SEVENTH EDITION

Microbiology
A Human Perspective
Eugene W. Nester
U N IVE RSIT Y OF W AS HI NGTO N

Denise G. Anderson
U N IVE RSIT Y OF W AS HI NGTO N

C. Evans Roberts, Jr.

U N IVE RSIT Y OF W AS HI NGTO N

Martha T. Nester

TM
 TM

MICROBIOLOGY: A HUMAN PERSPECTIVE, SEVENTH EDITION

Published by McGraw-Hill, a business unit of The McGraw-Hill Companies, Inc., 1221 Avenue of the Americas,
New York, NY 10020. Copyright © 2012 by The McGraw-Hill Companies, Inc. All rights reserved. Printed in the
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1 2 3 4 5 6 7 8 9 0 QDB/QDB 1 0 9 8 7 6 5 4 3 2 1

ISBN 978–0–07–337531–1
MHID 0–07–337531–4

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Library of Congress Cataloging-in-Publication Data

Nester, Eugene W.
Microbiology : a human perspective / Eugene Nester, Denise Anderson, C. Evans Roberts, Jr. ; contributors,
Deborah Allen, Sarah Salm. — 7th ed.
p. ; cm.
Includes index.
ISBN 978–0–07-337531–1 — ISBN 0–07–337531–4 (hard copy : alk. paper) 1. Microbiology. I. Anderson,
Denise G. (Denise Gayle). II. Roberts, C. Evans. III. Title.
[DNLM: 1. Microbiological Techniques. 2. Communicable Diseases—microbiology. QW 4]
QR41.2.M485 2012
616.9'041—dc22
2011010352
www.mhhe.com
Brief Contents
PAR T I P AR T IV
Life and Death of Microorganisms Infectious Diseases
1 Humans and the Microbial World 1 21 Respiratory System Infections 483
2 The Molecules of Life 17 22 Skin Infections 521
3 Microscopy and Cell Structure 39 23 Wound Infections 548
4 Dynamics of Prokaryotic Growth 82 24 Digestive System Infections 571
5 Control of Microbial Growth 107 25 Genitourinary Tract Infections 611
6 Metabolism: Fueling Cell Growth 126 26 Nervous System Infections 641
7 The Blueprint of Life, from DNA to 27 Blood and Lymphatic Infections 670
Protein 161 28 HIV Disease and Complications of
8 Bacterial Genetics 188 Immunodeficiency 694
9 Biotechnology and Recombinant DNA 215

P AR T V
PAR T I I Applied Microbiology
The Microbial World 29 Microbial Ecology 718
10 Identification and Classification of 30 Environmental Microbiology: Treatment of
Prokaryotic Organisms 237 Water, Wastes, and Polluted Habitats 735
11 The Diversity of Prokaryotic Organisms 255 31 Food Microbiology 748
12 The Eukaryotic Members of the Microbial
World 282 APPENDICES A–1

13 Viruses, Viroids, and Prions 304 GLOSSARY G–1

CREDITS C–1
INDEX I–1
PAR T I I I
Microorganisms and Humans
14 The Innate Immune Response 334
15 The Adaptive Immune Response 354
16 Host-Microbe Interactions 380
17 Immunologic Disorders 401
18 Applications of Immune Responses 419
19 Epidemiology 437
20 Antimicrobial Medications 457

iii
About the Authors
Eugene Nester Outside of academic life, Denise relaxes in the Phinney Ridge
neighborhood of Seattle, where she lives with her husband, Richard
Eugene (Gene) Nester performed
Moore, and dog, Dudley (neither of whom are well trained). When not
his undergraduate work at Cornell
planning lectures, grading papers, or writing textbook chapters, she can
University and received his Ph.D.
usually be found chatting with the neighbors, fighting the weeds in her
in microbiology from Case Western
garden, or enjoying a fermented beverage at the local pub.
University. He then pursued post-
doctoral work in the Department of
Genetics at Stanford University with
Joshua Lederberg. Since 1962, Gene C. Evans Roberts, Jr.
has been a faculty member in the Evans Roberts was a mathemat-
Department of Microbiology at the University of Washington. Gene’s ics student at Haverford College
research has focused on gene transfer systems in bacteria. His laboratory when a chance encounter landed
demonstrated that Agrobacterium transfers DNA into plant cells, the basis him a summer job at the Marine
for the disease crown gall. He continues to study this unique system of Biological Laboratory in Woods
gene transfer, which has become a cornerstone of plant biotechnology. Hole, Massachusetts. There, interac-
In 1990, Gene Nester was awarded the inaugural Australia Prize tions with leading scientists awak-
along with an Australian and a German scientist for their work on ened an interest in biology and
Agrobacterium transformation of plants. In 1991, he was awarded the medicine. After finishing his degree
Cetus Prize in Biotechnology by the American Society of Microbiology. at Haverford, he went on to get an M.D. degree at Columbia University
He has been elected to Fellowship in the National Academy of Sciences, College of Physicians and Surgeons, complete an internship at University
the American Academy for the Advancement of Science, the American of Rochester School of Medicine and Dentistry, and a residency in
Academy of Microbiology, and the National Academy of Sciences in medicine at University of Washington School of Medicine, where he
India. Throughout his career, Gene has been actively involved with the also completed a fellowship in infectious diseases under Dr. William
American Society for Microbiology in several leadership positions. M. M. Kirby, and a traineeship in diagnostic microbiology under Dr.
In addition to his research activities, Gene has taught an introductory John Sherris.
microbiology course for students in the allied health sciences for many Subsequently, Dr. Roberts taught microbiology at the University
years. He wrote the original version of the present text, Microbiology: of Washington, University of Oregon, and Chiang Mai University, in
Molecules, Microbes and Man, with C. Evans Roberts, Brian McCarthy, Chiagmai, Thailand, returning to the University of Washington thereafter.
and Nancy Pearsall more than 30 years ago because they felt no suitable He has directed diagnostic medical microbiology laboratories, served on
text was available for this group of students. The original text pioneered hospital infection control committees, and taught infectious diseases to
the organ system approach to the study of infectious disease. nurse practitioners in a camp for Karen refugees in Northern Thailand.
Gene enjoys traveling, museum hopping, and the study and collect- He has had extensive experience in the practice of medicine as it relates
ing of Northwest Coast Indian Art. He and his wife, Martha, live on Lake to infectious diseases. He is certified both by the American Board of
Washington with their labradoodle, Twana, and a well-used kayak. Their Microbiology and the American Board of Internal Medicine.
two children and four grandchildren live in the Seattle area. Evans Roberts worked with Gene Nester in the early development
of Microbiology: A Human Perspective. His professional publications
concern susceptibility testing as a guide to treatment of infectious diseases,
Denise Anderson etiology of Whipple’s disease, group A streptococcal epidemiology, use of
Denise Anderson is a Senior Lecturer fluorescent antibody in diagnosis, bacteriocin typing, antimicrobial resis-
in the Department of Microbiology tance in gonorrhea and tuberculosis, Japanese B encephalitis, and rabies.
at the University of Washington, For relaxation, he enjoys hiking, bird watching, and traveling worldwide.
where she teaches a variety of courses
including general microbiology,
recombinant DNA techniques, medical Martha Nester
bacteriology laboratory, and medical Martha Nester received an undergrad-
mycology/parasitology laboratory. uate degree in biology from Oberlin
Equipped with a diverse educational College and a Master’s degree in
background, including undergraduate education from Stanford University.
work in nutrition and graduate work in food science and in microbiology, She has worked in university research
she first discovered a passion for teaching when she taught microbiol- laboratories and has taught elemen-
ogy laboratory courses as part of her graduate training. Her enthusiastic tary school. She currently works in an
teaching style, fueled by regular doses of Seattle’s famous caffeine, environmental education program at
receives high reviews by her students. the Seattle Audubon Society. Martha

iv
 About the Authors v

has worked with her husband, Gene, for more than 40 years on microbiol- its treatment with bacteriophage; intracellular pathogens in general, their
ogy textbook projects, at first informally as an editor and sounding board, mechanism of pathogenicity, and the immune response to pathogen chal-
and then as one of the authors of Microbiology: A Human Perspective. lenge; and molecular and evolutionary immunology.
Martha’s favorite activities include spending time with their four grand-
Sarah Salm is a Professor at the Borough of Manhattan Community
children, all of whom live in the Seattle area. She also enjoys playing the
College (BMCC) of the City University of New York, where she
cello with a number of musical groups in the Seattle area.
teaches microbiology, anatomy and physiology, and general biology.
She earned her undergraduate and doctoral degrees at the University of
the Witwatersrand in Johannesburg, South Africa. She later moved to
About the Contributors New York, working first as a postdoctoral fellow and then an Assistant
Deborah Allen is an associate professor at Jefferson College in Missouri, Professor of Research at NYU Langone Medical Center. Her research
where she teaches microbiology as well as several other courses for has covered a range of subjects, from plant virus identification through
students entering allied health careers. Her graduate work was in zool- prostate stem cell characterization. She left her research position at NYU
ogy at the University of Oklahoma and in neurobiology and behavior at to focus exclusively on her position at BMCC, where she has been enthu-
Cornell University. She participated in cancer research at the University siastically teaching since 2004.
of Arkansas Medical Center before embarking on a career in publishing,
Teri Shors is a Professor in the Department of Biology and
acquiring and developing books in the life sciences. She is now thrilled to
Microbiology at the University of Wisconsin–Oshkosh. Teri began a
be working on the other end of the desk with the Nester team. Away from
tenure track position in the Department of Biology and Microbiology
campus, Deborah reads or listens to her favorite Eve Dallas novels, floats
in September 1997. Prior to her arrival at UW–Oshkosh, she served
the rivers and listens to folk music in the Ozarks, and fully appreciates the
two years as a postdoctoral fellow at the National Institutes of Health,
local microbes while visiting Missouri wineries.
in Bethesda, Maryland, under the direction of Dr. Bernard Moss, Chief
Nancy Boury is a Senior Lecturer at Iowa State University, where she also of the Laboratory of Viral Diseases, National Institute of Allergies and
received her Doctorate of Molecular Biology. She received her Master’s Infectious Diseases. Her research training/specialty has been in the field
degree from the University of Wisconsin–Madison in medical microbiol- of poxviruses. The courses she teaches center around laboratory and
ogy. She currently teaches biology, microbiology, and genetics courses. lectures in microbiology, virology, molecular and cellular biology, and
Nancy is developing a mouse model for oral challenge salmonellosis and graduate seminar.

DE DI CATION

We dedicate this book to our students;

we hope it helps to enrich their lives and
to make them better-informed citizens,

to our families
whose patience and endurance
made completion of this project a reality,

to Anne Nongthanat Panarak Roberts

in recognition of her invaluable help,
patience, and understanding, and

to our colleagues
for continuing encouragement
and advice.
 n e N e s t e r
f r o m G e
Letter

s and S t u d ents: step back f

rom
Instru c t o r , it’s time f o r m e t o
erson,
n Denise And
its 7 editio
th
o k e n t e r s o n s u lt a n t .
As this text
bo ior c
it e r t o b e c oming a sen t a k e on increasin
g
tiv e w r ditio n , w il l
being an ac ce the 2 e ember of th
e
nd
a u t h o r s in e c o m e a m
en an to b
who has be it ie s . I e n li sted Denise ic r o b iology, had
a
resp on s ib il love f o r m
authorship
s e s h e h a d an obvious n d w a s a n enthusiastic
becau gy, a
author team ic robial biolo the
g in a ll a spe c t s o f m
v e y e d h e r passion for
in con
broad train r w h o successfully r m e m b ership on
tea c h e nts . H e
and dynamic r u n d e r g r a duate stude t h e success of t
he
orld t o h e u s ly t o
microbial w ormo will enhanc
e
r s h a s c o n tributed en o n f id e n c e
f autho yc
the team o it io n s a n d I have ever lm , this textboo
k
p a s t e d t t h e h e
ell as . With Denis
ea
present as w r e e d it io n s s t u d e nts o f the allied
of all futu non-majors
and
the quality st c h o ic e f o r
the very be
will remain G e n e Nester
n c e s . —
health scie

vi
Customize Your Perfect Foundation

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vii
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viii
and Students to Course Concepts

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ix
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Animations Over 100 animations bringing key concepts to life,
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xi
ALL NEW Instructional Art Program
Every piece of art has been revised or replaced to make it more vibrant, three-dimensional,
and instructional. The goal of this extensive update was to evaluate every piece, give
colors and shapes consistency throughout, and make all art more engaging and useful
(for students and instructors). A more modern, instructional art program helps students
understand key concepts.

Budding of an Enveloped Virus

As you can see, the seventh edition figure is much more instructive. The student
will have a much easier time understanding how this process works! 6e
7e

FIGURE 13.15a

FIGURE 14.6a

Replication/Transcription/Translation
6e
Focus Figures highlight key topics.
7e

FIGURE 7.1

FIGURE 7.1

xii
System Overviews
The system chapters have anatomically correct figures
to help the students throughout the chapter.
7e
6e

FIGURE 22.2b

FIGURE 21.1b

Colors are more vibrant and figures

have a much more 3D effect.
7e
6e

FIGURE 3.24

FIGURE 3.24

xiii
LAYING THE FOUNDATION
Clarified Writing Helps Students Focus on Learning the Science
Microbiology is one of the fastest-moving fields, so it is not enough to teach students only the current hot topics. Students must be armed
with a strong foundation to understand the next year’s—even the next decade’s—hot topics as well. The Nester author team works hard,
both in their textbook and in their classrooms, to teach students for the future, as well as for today. Microbiology: A Human Perspective
remains focused on providing an excellent foundation in fundamental concepts for microbiology students of all backgrounds. With this
edition, the authors underwent the tremendous task of combing through the narrative with a critical eye—for example, general vocabulary
and reading level were simplified so that students can focus on learning about microbiology and the complex terminology required. The
seventh edition of Microbiology: A Human Perspective incorporates the changes necessary to ensure it is readable for students—easier to
comprehend—yet still maintains an appropriately rigorous level of science.

Table of Contents has been updated as necessary to

improve comprehension, reduce redundancy, and help students
build a solid foundation in microbiology.

NEW!
Key Terms begin every chapter to start building Learning Outcomes begin each major section. They are
a foundation! numbered to correspond to digital materials.

xiv
 MicroAssessments
Major sections end with a MicroAssessment that summarizes the
major concepts in that section and offers both review questions
and critical-thinking questions (marked with a + ) to assess
understanding of the preceding section.

NEW!
Figure Questions offer
another assessment opportunity
for students. Most figures have a
critical-thinking question associated
with them. ? Animations found
online correlate to many images
within the text.

End of Chapter Review

• Short Answer questions review major chapter concepts.
• Multiple Choice questions allow self-testing; answers are provided in
Appendix IV.
• Applications provide an opportunity to use knowledge of microbiology to
solve real-world problems.
• Critical Thinking questions encourage practice in analysis and problem
solving that can be used by the student in any subject.

MicroBytes
Found throughout the chapters, these are
small “bytes” of interesting information.

xv
BUILDING UP THE BASE
A Glimpse of History
Each chapter opens with an engaging story about the men and
women who pioneered the field of microbiology.

Perspective Boxes
Perspective boxes introduce a
human perspective by showing
how microorganisms and their
products influence our lives in
many different ways.

Future Challenges
Many chapters end with a pending challenge
facing current and future microbiologists.

Case Presentations
Each infectious disease chapter includes a case presentation
of a realistic clinical situation.

xvi
 Unmatched Clinical Coverage
Organized by human body systems, the infectious disease chapters
(21 to 28) are highlighted with purple shading in the top corner of
the page for easy reference.

Incomparable Presentation
of Diseases
Each disease is presented systematically and
predictably. Individual sections describe the
disease’s symptoms, causative agents, pathogenesis,
epidemiology, and prevention and treatment.

IMPROVED!
Enhanced Disease Summaries
Major diseases are represented with a summary table that
includes an outline of pathogenesis keyed to a human
figure showing the entry and exit of the pathogen. NEW!
Diseases in
Review Table
Each infectious
disease chapter
ends with a table
that summarizes the
key features of the
diseases discussed in
that chapter.

xvii
UPDATES—Enhancing the Foundation
Global Changes Chapter 3
■ All the art was revised to make it more engaging and clear ■ New figure that shows permeability of the lipid bilayer of the
cytoplasmic membrane (figure 3.25a)
■ Added diversity to the “disease person” in the disease sum-
mary tables, using different “people” rather than a generic ■ New figure that shows aquaporins (figure 3.25b)
“person” ■ Combined the figures that show types of transport systems
■ New feature—MicroBytes—short snippets of information, (figure 3.29)
usually 140 characters or less ■ New figure on secretion (figure 3.30)
■ New feature—Questions for most figures are meant to engage ■ New section on osmosis
students in the figure and encourage students to read the ■ Added information about the gel-like material sandwiched
accompanying information in the narrative between the cytoplasmic membrane and the Gram-positive
■ New feature—Diseases in Review tables highlight key cell wall
characteristics of diseases discussed in the chapter, helping ■ Added information on multiphoton microscopes
students to focus on the “big picture”
■ New feature—Focus Figures emphasize the key points of the Chapter 4
general topic of the chapter ■ New figure that shows the development of a biofilm (figure 4.3)
■ In the disease chapters, emphasized the ecology of the organ ■ New section on reactive oxygen species (ROS)
system by combining the previous sections “Anatomy and
Physiology” and “Normal Microbiota,” renaming the single Chapter 5
section “Anatomy, Physiology, and Ecology”
■ Added term “sporocide”
■ Decreased the number of bolded key terms—this was done
so that students focus more on the explanations, not just the
■ Added information on EPA’s “designed for the environment”
terms. Note that most of the previously bolded terms are still label
in the text; students can highlight these terms if they want
Chapter 6
■ Most bolded terms are in the glossary
■ Added a figure to show that many glucose molecules enter a
■ Tightened and simplified the explanations and descriptions
cell, and they can have different fates (figure 6.9)
throughout the text, with the aim of making the most important
information stand out; overall, the information is easier to read ■ Added colored icons to the table that shows precursor mol-
ecules (table 6.2), and these have been incorporated in other
Key Changes in Individual figures in place of the previous gray boxes
Chapters ■ Moved the figure that shows the effect of the energy source
versus the terminal electron acceptor forward in the chapter,
Chapter 1 and used it to give examples (figure 6.7)
■ Combined two sections to create one: “The Dispute Over ■ Changed the topic of the “Future Challenges” story to biofuels
Spontaneous Generation”
■ Added “The Golden Age of Microbiology” to the timeline Chapter 7
(figure 1.2) ■ New figure to show three kinds of RNA involved in protein
synthesis (figure 7.3)
Chapter 2 ■ New figure to show amplification with transcription/
■ Updated depiction of atoms to match that used in chemistry translation (figure 7.4)
texts, using only Lewis figures to show the number of elec- ■ New figure to show the regions on DNA that direct transcrip-
trons in the valence shell (figures 2.1 to 2.4) tion (figure 7.8)
■ Updated the depiction of polar compounds by using electron ■ New figure to show the regions on mRNA that direct transla-
density molecules (figures 2.5 and 2.6) tional (figure 7.12)
■ Introduced the term “electronegativity” (table 2.2) ■ New figure that uses an analogy to show the principles of
■ Added a table to illustrate biologically important functional regulation (figure 7.20)
groups (table 2.4) ■ Reordered the sections on regulation, so that sensing mecha-
■ Added a section on protein domains nisms are covered first.
xviii
■ Added inducer exclusion to the mechanisms of carbon catab- Chapter 13
olite repression
Viruses, Viroids, and Prions (Note that chapter titles are included
from this point on because of the change in the Table of Contents.)
Chapter 8
■ Condensed the information previously spread over two chap-
■ New figure that shows the mutagenic effects of an alkylating ters into one (to do this, overlapping information, including
agent (figure 8.7) details that overlapped with coverage in the disease chapters,
was eliminated)
Chapter 9 ■ New section on CRISPR, including a figure (figure 13.11)
■ Updated by no longer italicizing restriction enzyme names
■ Introduced the term “high throughput” Chapter 14
■ Added a MicroByte about the Genetic Information Non- The Innate Immune Response
discrimination Act (GINA) ■ Added an overview figure (figure 14.1)
■ Added information about the successful replacement of a ■ Highlighted the information on pattern recognition recep-
bacterial chromosome by a machine-made chromosome. tors by creating a new section (14.5); added information on
danger-associated molecular patterns (DAMPs), inflamma-
Chapter 10 somes, and RIG-like receptors
■ Updated taxonomic outline of Bergey’s Manual of Systematic ■ Added a small section on regulation of the complement sys-
Bacteriology (table 10.3) tem to prepare students for understanding how some patho-
gens hijack that mechanism to avoid activating the system
■ Moved a figure on serotypes from chapter 11 (now figure 10.9)
■ Added a section on the damaging effects of the inflamma-
■ Added a section on nucleic acid amplification tests (NAATs),
tory response to prepare students for understanding how the
incorporating the previous information on using PCR for
response can both protect against disease and also cause dis-
identification
ease symptoms
■ Added information on multilocus typing to the section on
■ Added a section on cell death and the inflammatory response
characterizing strain differences using molecular typing (pre-
to explain the difference between cell death due to direct
viously titled “Genomic Typing”)
damage, apoptosis, and pyroptosis
Chapter 11
Chapter 15
■ Updated to use the term “endoflagella” instead of axial
The Adaptive Immune Response
filaments
■ Overview figure has been incorporated as a “mini-figure” into
■ Added information about the problem associated with other figures
Coxiella in the placenta of infected animals
■ New figure showing a Peyer’s patch (figure 15.5)
■ Modified figures of B-cell activation and T-cell activation to
Chapter 12 show that lack of accessory signals induces anergy, thereby
■ Rearranged the order of the chapter so that fungi, which more promoting tolerance (figures 15.11 and 15.20)
students are probably familiar with, are covered first, fol- ■ New figure showing natural killer (NK) cells destroying
lowed by algae and protozoa “stressed” cells that lack MHC class I molecules (figure 15.23)
■ Updated classification of fungi
■ Updated classification of protozoa Chapter 16
■ Updated classification of eukaryotes (figures 12.10 and 12.13) Host-Microbe Interactions
■ New photo of water mold (figure 12.16) ■ Shortened figure that shows mechanisms viruses use to avoid
detection by the MHC class I antigen presentation (figure 16.13)
■ Separated the section that previously combined the coverage
to avoid overlap with the new figure in chapter 15
of arthropods and helminths and switched the order
■ New photo of elephantiasis (figure 12.17) Chapter 17
■ New life cycle figure of ascariasis (figure 12.18)
Immunologic Disorders
■ Created consistent subheadings within the sections on fungi, ■ New photo of a goiter (figure 17. 9)
algae, and protozoa to cover types, structure, habitats, repro-
duction, and economic or medical importance
Chapter 18
■ New box on river blindness (Perspective 12.1)
Applications of Immune Responses
■ Expanded coverage of insect vectors (table 12.6) ■ New figure that shows the host-pathogen “trilogy”—the
■ New photo of Anopheles mosquito, vector of malaria immune wars, the microbes fight back, and the return of the
(figure 12.20) humans (figure 18.1)
xix
 ■ Added a section on the basic principles of using labeled Chapter 23
antibodies, including a figure that highlights the difference
Wound Infections
between direct and indirect tests (figure 18.9)
■ Reordered coverage in “Bacterial Infections of Bite Wounds”
■ Moved the information on radial immunodiffusion and immu- so that “Human Bites” comes first, reflecting the student
noelectrophoresis to the Web, thereby placing more emphasis population’s interest in the topic
on the other techniques covered
■ Eliminated some of the overlap in the coverage of Staphylo-
coccus aureus in chapters 22 and 23, referring the reader back
Chapter 19 to chapter 22
Epidemiology
■ New figure on vector transmission, showing the action of a Chapter 24
mechanical vector and a biological vector (figure 19.6)
Digestive System Infections
■ Split a previous section into two: “Pathogen Factors That Influ- ■ Added a new figure to highlight the difference between infec-
ence the Epidemiology of Infectious Disease” and “Host Fac- tions of the teeth and infections of the gums (figure 24.3)
tors That Influence the Epidemiology of Infectious Disease”
■ Updated the information on periodontal diseases, emphasiz-
■ Created a separate primary section on “Emerging Infectious ing the polymicrobial nature of the disease and the role of the
Diseases” inflammatory response in the disease process
■ Updated terminology introducing case-fatality rate ■ Added a new section “General Characteristics of Diarrheal
■ Separated one figure into three separate figures showing Diseases” to highlight the importance of oral rehydration
(1) reservoirs of infection (figure 19.2), (2) portals of entry therapy and other overlapping concepts related to these dis-
(figure 19.3), and (3) mechanisms of transmission (figure 19.4) eases; a new figure (figure 24.10) summarizes the common
■ Combined portals of exit and portals of entry into single section mechanisms of pathogenesis of intestinal pathogens
■ Moved the information on Clostridium difficile, highlighting
Chapter 20 it in a new section: “Clostridium difficile–Associated Disease
(CDAD)”; added a photo of pseudomembranous colitis (fig-
Antimicrobial Medications ure 24.14)
■ New section that describes carbapenem-resistant Enterobac-
■ Updated the information on cholera to include its recent intro-
teriaceae (CRE)
duction into Haiti
■ New section on integrase inhibitors, used to treat HIV infection
■ Updated the information on E. coli gastroenteritis to empha-
size the different virulence factors and epidemiology of the
Chapter 21 different pathovars
Respiratory System Infections ■ Added paratyphoid fever to the section on typhoid fever
■ New section “Post-Streptococcal Sequelae” added to empha- ■ Separated viral diseases of the lower digestive system into
size the complications that can follow strep throat two sections: those that cause intestinal diseases and those
■ Increased coverage of the signs and symptoms of whooping that cause liver diseases
cough to emphasize the three stages; also added information ■ Increased the coverage on hepatitis B virus antigens to
about the recent outbreak in California emphasize their clinical significance
■ Updated coverage of tuberculosis, adding the terms “latent
tuberculosis infection (LTBI)” and “active tuberculosis dis-
Chapter 25
ease (ATBD)”; added a new figure to illustrate the pathogen-
esis of tuberculosis (figure 21.19) Genitourinary Tract Infections
■ Updated by substituting the term “sexually transmitted infec-
■ Updated the coverage of influenza, emphasizing how anti-
tion (STI)” in place of “sexually transmitted disease (STD)”
genic drift is responsible for seasonal influenza, and antigenic
to reflect that many of the infections do not cause noticeable
shift is responsible for pandemic influenza
disease
■ Added Weil’s disease to the coverage of leptospirosis
Chapter 22
■ Added “Latent Syphilis” to the stages of the disease
Skin Infections
■ New photograph of the bull’s-eye rash of Lyme disease
(figure 22.10) Chapter 26
■ New photograph of shingles (figure 22.16) Nervous System Infections
■ Moved the information on pneumococcal meningitis, Hae-
■ New section “Acne Vulgaris” on acne
mophilus influenzae meningitis, and neonatal meningitis into
■ Moved the information on Malassezia furfur and Candida albi- three separate new sections, thereby increasing the emphasis
cans, highlighting it in a new section “Other Fungal Diseases” on these diseases

xx
■ New figure showing meningitis belt in Africa (figure 26.5) ■ Added information about Plasmodium knowlesi to the malaria
■ New figure that highlights the use of bacteriophage prepara- section
tions to decrease the risk of listeriosis associated with ready- ■ Altered the coverage of plague to emphasize the importance
to-eat foods (figure 26.7) on the route of entry of Yersinia pestis in the disease symp-
■ New photo showing early signs of leprosy (figure 26.8a) toms and outcome
■ New graph showing the incidence of the three types of botu-
lism in the United States (figure 26.11) Chapter 28
■ Updated the information on Cryptococcus gattii; added map HIV Disease and Complications of Immunodeficiency
of its spread in the Pacific Northwest (figure 26.18) ■ Incorporated more structure in the coverage of HIV/AIDS by
■ Added a section on “Primary Amebic Meningoencephalitis subdividing the information further
(PAM)” ■ Added information on the role of viral set point in HIV dis-
■ Updated box on rabies survivors ease progression
■ Updated the information on HIV prevention by adding infor-
mation on male circumcision and new vaccine candidates
Chapter 27
Blood and Lymphatic Infections
■ Updated the coverage of sepsis and septic shock to emphasize Chapter 29
the role of the inflammatory response in the disease process Microbial Ecology
■ New section on “Dengue Fever” ■ Revised figures of biogeochemical cycles to highlight the
role that the reaction plays in microbial growth (biosynthesis,
■ New section on “Chikungunya”
energy source, terminal electron acceptor)
■ New section on “Emerging Hemorrhagic Fevers” (Ebola and
Marburg)

xxi
 microbiology

Unique Interactive Question Types Pre-tagged to

ASM’s Curriculum Guidelines for Undergraduate Microbiology

1. Case Study: Case study questions use tutorial and assessment to help students see the world of microbiology
and how it applies to their lives. These static case studies come to life in a learning activity that is interactive, self-
grading, and assessable. The integration of the cases with videos and animations enhances the depth of the content
covered, and the use of integrated questions forces students to stop, think, and evaluate their understanding. Pre- and
post-testing allow instructors and students to assess the overall comprehension of the activity.

2. System Summary Figures: System overview figures from the text are made interactive to give students addi-
tional opportunity to practice and assess their knowledge. Key components of each disease can be reviewed includ-
ing signs and symptoms, causative agent, incubation period, epidemiology, and more.

3. Key Concept Activities: These learning modules take students deep into the key microbiology concepts they
need to know for long-term success in the course. Pre- and post-assessment is used in addition to multimedia tutori-
als to ensure students gain mastery of the key topics in each chapter.

4. Disease Pathway Figures: This question correlates to the Diseases In Review table found at the end of each
disease chapter and will allow students to practice on the most important information to remember for each disease
covered in the chapter.

5. Tutorial Animation Learning Modules: Making use of McGraw-Hill’s collection of videos and animations,
this new question type presents an interactive, self-grading, and assessable activity. Pre- and post-testing is used to
assess student comprehension and the use of integrated questions forces students to stop, think, and evaluate their
understanding of the process being presented. These tutorials take a stand-alone, static animation and turn it into an
interactive learning experience for your students with real-time remediation.

6. Labeling: Using the high-quality art from the textbook, check your students’ visual understanding as they practice
interpreting figures and learning structures and relationships. Easily edit or remove any label you wish!

7. Classification: Ask students to organize concepts or structures into categories by placing them in the correct
“bucket.”

8. Sequencing: Challenge students to place the steps of a complex process in the correct order.

9. Composition: Fill in the blanks to practice vocabulary, and then reorder the sentences to form a logical paragraph
(these exercises may qualify as “writing across the curriculum” activities!)

All McGraw-Hill ConnectPlus® content is pre-tagged to Learning Outcomes for each chapter as well as topic, section,
Bloom’s Level, and ASM Curriculum Guidelines (once they’re finalized) to assist you in both filtering out unneeded
questions for ease of creating assignments and in reporting on your students’ performance against these points. This
will enhance your ability to effectively assess student learning in your courses by allowing you to align your learning
activities to peer-reviewed standards from an international organization.
Acknowledgments
Gene Nester, Evans Roberts, Brian McCarthy, and Nancy Pearsall Erica Perrer, Delgado Community College
shared a vision many years ago to write a new breed of microbi- Madhura Pradhan, Ohio State University
ology textbook especially for students planning to enter nursing Dr. Marceau Ratard, Delgado Community College
and other health-related careers. Today there are other books of Pele Eve Rich, North Hennepin Community College
this type, but we were extremely gratified to learn that a major- Seth Ririe, Brigham Young University–Idaho
ity of the students we surveyed intend to keep their copies of Deemah N. Schirf, University of Texas at San Antonio
Microbiology: A Human Perspective because they feel it will George F. Spiegel, Jr., College of Southern Maryland
benefit them greatly as they pursue their studies in these fields. Steve Thurlow, Jackson Community College
Special thanks to the many students who used Microbiology: A Floyd L. Wormley, Jr., University of Texas at San Antonio
Human Perspective over the years and who shared their thoughts Malcolm Zellars, Georgia State University
with us about how to improve the presentation for the students
who will use this edition of the text.
We offer our sincere appreciation to the many gracious and
expert professionals who helped us with this revision by offer-
Focus Group Participants
ing helpful suggestions. In addition to thanking those individuals Corrie Andries, Central New Mexico Community College
listed here who carefully reviewed chapters, we also thank those John Bacheller, Hillsborough Community College
who responded to our information surveys, those who participated Michelle Badon, University of Texas at Arlington
in regional focus groups, and those participants who chose not David Battigelli, University of North Carolina–Greensboro
to be identified. All of you have contributed significantly to this Nancy Boury, Iowa Sate University
work and we thank you. Lance Bowen, Truckee Meadows Community College
William Boyko, Sinclair Community College
Dave Brady, Southwestern Community College
Antonia Brem, Wayne County Community College
Reviewers of the Seventh Edition Chad Brooks, Austin Peay State University
Ahmmed Ally, Massachusetts College of Pharmacy and Health Lisa Burgess, Broward College
Sciences Liz Carrington, Tarrant Country College–Southeast Campus
Cindy B. Anderson, Mt. San Antonio College Joseph Caruso, Florida Atlantic University
James Barbaree, Auburn University Erin Christensen, Middlesex Community College
Kris Baumgarten, Delta College Robin Cotter, Phoenix College
Suzanne D. Clutter, West Virginia Northern Community College– John Dahl, Washington State University
Wheeling Alison Davis, East Los Angeles College
Lorraine A. Cramer, University of North Carolina at Chapel Hill Ana Dowey, Palomar College
Thomas R. Danford, West Virginia Northern Community College Elizabeth Emmert, Salisbury University
Judy Earl, Montgomery County Community College Susan Finazzo, Georgia Perimeter College
Kathryn Germain, Southwest Tennessee Community College Clifton Franklund, Ferris State University
Robert Gessner, Valencia Community College Jason Furrer, University of Missouri
Julie Gibbs, College of DuPage Chris Gan, Highline Community College
Amy D. Goode, Illinois Central College Edwin Gines-Candalaria, Miami–Dade
Ellen J. Gower, Greenville Technical College Zaida Gomez-Kramer, University of Central Arkansas
Judy Haber, California State University Fresno Amy Good, Illinois Central College
John Ireland, Jackson Community College Todd Gordon, Kansas City Kansas Community College
Shirley E. Kangas, Cuyahoga Community College, Brinda Govindan, San Francisco State University
Eastern Campus Julianne Grose, Brigham Young University
Amine Kidane, Columbus State Community College Gabriel Guzman, Triton College
P. Kourtev, Central Michigan University Judy Haber, California State University–Fresno
Timothy A. Kral, University of Arkansas Julie Harless, Lone Star College
Judith Marie Krey, Waubonsee Community College Zafer Hatahet, Northwestern State University
William Lorowitz, Webster State University James Herrick, James Madison University
Jennifer Leigh Myka, Galen College of Nursing–Cincinnati Jennifer Herzog, Herkimer County Community College
Karen Nakaoka, Webster State University Dena Johnson, Tarrant County College
Steven Obenauf, Broward College Jim Johnson, Central Washington University

xxiii
xxiv Acknowledgments

Eunice Kamunge, Essex County College over many years. Our special thanks go to John Leigh, Mary Bicknell,
Amine Kidane, Columbus State Community College Mark Chandler, Kendall Gray, Janis Fulton, Jimmie Lara, Sharon
Jeff Kiggins, Monroe Community College Schultz, Michael Lagunoff, Heather Felise, Julie Overbaugh, and
Terri Lindsey, Tarrant County College James Staley for their general suggestions and encouragement.
Suzanne Long, Monroe Community College We would also like to thank Denise’s husband, Richard
Kimberly Maznicki, Seminole State College of Florida Moore, who was “forced” to proofread and critique many of the
Caroline McNutt, Schoolcraft College chapters. Although he has no formal scientific education, or per-
Elizabeth McPherson, University of Tennessee–Knoxville haps because of that fact, his suggestions have been instrumental
Amee Mehta, Seminole State College of Florida in making the text more “reader-friendly.” Much to his own
Sharon Miles, Itawamba Community College surprise, Richard has learned enough about the fundamentals of
Tracey Mills, Ivy Tech Community College–Lawrence microbiology to actually become intrigued with the subject.
Rita Moyes, Texas A & M University Special thanks to the reviewers and other instructors who
Valerie Narey, Santa Monica College helped guide us in this revision. Deciding what to eliminate, what
Ruth Negley, Harrisburg Area Community College to add, and what to rearrange is always difficult, so we appreciate
Steven Obenauf, Broward College your input.
Julie Oliver, Cosumnes River College Thanks also to Jason Overby, who provided guidance as we
Marcia Pierce, Eastern Kentucky University updated our chemistry coverage in this edition, and David Hurley,
Madhura Pradhan, Ohio State University who helped us navigate the murky waters during a substantial
Todd Primm, Sam Houston State University revision of an earlier edition that updated the coverage of innate
Jean Revie, South Mountain Community College and adaptive immunity.
Jackie Reynolds, Richland College A list of acknowledgments is not complete without thanking
Beverly Roe, Erie Community College our fearless leaders from McGraw-Hill. Our sponsoring editor
Silvia Rossbach, Western Michigan University Lynn Breithaupt and developmental editor Fran Simon not only
Benjamin Rowley, University of Central Arkansas gave inspiration and sound advice, but laughed at our jokes and
Donald Rubbelke, Lakeland Community College became true friends. Our project manager Mary Jane Lampe and
Mark Schneegurt, Wichita State University copyeditor Bea Sussman were instrumental in making sure the
Teri Shors, University of Wisconsin–Oshkosh correct words actually made it onto paper.
Sasha Showsh, University of Wisconsin–Eau Claire Additionally, we would like to thank John Bacheller, Mira
Heidi Smith, Front Range Community College Beins, Chad Brooks, Liz Carrington, Greg Frederick, Renee
Louise Thai, University of Missouri–Columbia Krohne, Jonathan Miller, Bethany Morgan, Julie Oliver, Seth
Steve Thurlow, Jackson Community College Ririe, Ben Rowley, and George Wawrzyniak for producing our
Sanjay Tiwary, Hinds Community College digital resources to support us and other instructors who lecture
Stephen Wagner, Stephen F. Austin State University from our text.
Delon Washo-Krupps, Arizona State University We hope very much that this text will be interesting, educa-
George Wawrzyniak, Milwaukee Area Technical College tional for students, a help to their instructors, and will convey the
Janice Webster, Ivy Tech Community College excitement that we all feel for the subject. We would appreciate
Mary Weintraub, University of Cincinnati any comments and suggestions from our readers.
John Whitlock, Hillsborough Community College
Samia Williams, Santa Fe Community College Eugene Nester
Fadi Zaher, Gateway Technical College Denise Anderson
C. Evans Roberts, Jr.
We thank our colleagues in the Department of Microbiology at the Martha Nester
University of Washington who have lent their support of this project
Contents
About the Authors iv ■ Preface vi

2.3 Chemical Components of the Cell 22

PAR T I
Water 22
Life and Death of Microorganisms Elements and Small Molecules in the Cell 24
Macromolecules and Their Component Parts 24
1 Humans and the Microbial World 1
2.4 Proteins 25
A Glimpse of History 1 Amino Acids 25
Key Terms 1 Peptide Bonds 27
Protein Structure 27
1.1 The Dispute Over
Protein Domains 28
Spontaneous Generation 1
Substituted Proteins 29
Early Experiments 2
Protein Denaturation 29
Experiments of Pasteur 2
Experiments of Tyndall 2 2.5 Carbohydrates 29
Monosaccharides 30
1.2 Microbiology: A Human Perspective 3
Disaccharides 31
Vital Activities of Microorganisms 3
Polysaccharides 31
Applications of Microbiology 3
Medical Microbiology 6 2.6 Nucleic Acids 32
Microorganisms as Model Organisms 8 DNA 32
RNA 33
1.3 Living Members of The Microbial World 8
Diversity of Microorganisms 8 2.7 Lipids 33
Bacteria 9 Simple Lipids 33
Archaea 9 Compound Lipids 35
Eucarya 10 Steroids 36
Nomenclature 11
PERSPECTIVE 2.1: Isotopes: Valuable Tools for the Study of
1.4 Non-Living Members of the Microbial World 11 Biological Systems 19
FUTURE CHALLENGES: Fold Properly: Do Not Bend or Mutilate 37
1.5 Size in the Microbial World 12
SUMMARY 36
PERSPECTIVE 1.1: Every Rule Has an Exception 14 REVIEW QUESTIONS 38
FUTURE CHALLENGES: Entering a New Golden Age 15

SUMMARY 15 3 Microscopy and Cell Structure 39

REVIEW QUESTIONS 16
A Glimpse of History 39
2 The Molecules of Life 17 Key Terms 39

MICROSCOPY AND CELL

A Glimpse of History 17
MORPHOLOGY
Key Terms 17
3.1 Microscopic Techniques:
2.1 Atoms and Elements 17 The Instruments 40
2.2 Chemical Reactions and Principles of Light Microscopy: The Bright-Field
Bonds 19 Microscope 40
Ions and Ionic Bonds 20 Light Microscopes That Increase Contrast 43
Covalent Bonds 20 Other Light Microscopes 44
Hydrogen Bonds 21 Electron Microscopes 44
Molarity 22 Atomic Force Microscopes 46

xxv
xxvi Contents

3.2 Microscopic Techniques: Dyes and Staining 46 3.12 Protein Structures Within the Cell 73
Differential Staining 46 Ribosomes 73
Special Stains to Observe Cell Structures 48 Cytoskeleton 73
Fluorescent Dyes and Tags 49 Flagella and Cilia 74
3.3 Morphology of Prokaryotic Cells 50 3.13 Membrane-Bound Organelles 74
Shapes 50 The Nucleus 74
Groupings 51 Mitochondria 74
Multicellular Associations 51 Chloroplasts 76
Endoplasmic Reticulum (ER) 76
THE PROKARYOTIC CELL
The Golgi Apparatus 77
3.4 The Cytoplasmic Membrane 52 Lysosomes and Peroxisomes 77
Structure of the Cytoplasmic Membrane 52
PERSPECTIVE 3.1: The Origins of Mitochondria and Chloroplasts 76
Permeability of the Lipid Bilayer 54
FUTURE CHALLENGES: A Case of Breaking and Entering 78
The Role of the Cytoplasmic Membrane in Energy
Transformation 55 SUMMARY 78
REVIEW QUESTIONS 80
3.5 Directed Movement of Molecules Across
the Cytoplasmic Membrane 55
Transport Systems 55
Protein Secretion 56
4 Dynamics of Prokaryotic Growth 82

3.6 Cell Wall 58 A Glimpse of History 82

Peptidoglycan 58 Key Terms 82
The Gram-Positive Cell Wall 58
4.1 Principles of Prokaryotic
The Gram-Negative Cell Wall 60
Growth 82
Antibacterial Substances That Target Peptidoglycan 62
Cell Wall Type and the Gram Stain 62 4.2 Prokaryotic Growth
Bacteria That Lack a Cell Wall 62 in Nature 84
Cell Walls of the Domain Archaea 62 Biofilms 84
Interactions of Mixed Microbial Communities 84
3.7 Capsules and Slime Layers 62
4.3 Obtaining a Pure Culture 85
3.8 Filamentous Protein Appendages 63
Growing Microorganisms on a Solid Medium 85
Flagella 63
The Streak-Plate Method 86
Pili 65
Maintaining Stock Cultures 87
3.9 Internal Structures 66
4.4 Prokaryotic Growth in Laboratory Conditions 87
The Chromosome 66
The Growth Curve 87
Plasmids 66
Colony Growth 88
Ribosomes 66
Continuous Culture 88
Cytoskeleton 66
Storage Granules 66 4.5 Environmental Factors That Influence Microbial
Gas Vesicles 67 Growth 89
Endospores 67 Temperature Requirements 89
Oxygen (O2) Requirements 90
THE EUKARYOTIC CELL pH 91
3.10 The Plasma Membrane 70 Water Availability 91

3.11 Transfer of Molecules Across the Plasma 4.6 Nutritional Factors That Influence Microbial
Membrane 72 Growth 92
Transport Proteins 72 Required Elements 92
Endocytosis and Exocytosis 72 Growth Factors 93
Secretion 73 Energy Sources 93
Nutritional Diversity 93
 Contents xxvii

4.7 Cultivating Prokaryotes in the Laboratory 94 PERSPECTIVE 5.1: Contamination of an Operating Room by a
Bacterial Pathogen 119
General Categories of Culture Media 94
Special Types of Culture Media 95 FUTURE CHALLENGES: Too Much of a Good Thing? 122
Providing Appropriate Atmospheric Conditions 96 SUMMARY 122
Enrichment Cultures 97 REVIEW QUESTIONS 124
4.8 Methods to Detect and Measure Microbial
Growth 98
Direct Cell Counts 98 6 Metabolism: Fueling Cell Growth 126
Viable Cell Counts 99
A Glimpse of History 126
Measuring Biomass 102
Key Terms 126
Detecting Cell Products 103
6.1 Principles of
PERSPECTIVE 4.1: Can Prokaryotes Live on Only Rocks and
Water? 94
Metabolism 127
Harvesting Energy 127
FUTURE CHALLENGES: Seeing How the Other 99% Lives 104
Components of Metabolic Pathways 128
SUMMARY 104 Precursor Metabolites 132
REVIEW QUESTIONS 105 Overview of Catabolism 132

5 Control of Microbial Growth 107 6.2 Enzymes 135

Mechanisms and Consequences of Enzyme Action 135
A Glimpse of History 107 Cofactors 136
Key Terms 107 Environmental Factors That Influence Enzyme
Activity 136
5.1 Approaches to Allosteric Regulation 137
Control 107 Enzyme Inhibition 138
Principles of Control 108
Situational Considerations 108 6.3 The Central Metabolic Pathways 139
Glycolysis 139
5.2 Selection of an Antimicrobial Procedure 110 Pentose Phosphate Pathway 141
Type of Microbes 110 Transition Step 141
Numbers of Microorganisms 110 Tricarboxylic Acid (TCA) Cycle 142
Environmental Conditions 111
Risk for Infection 111 6.4 Respiration 142
Composition of the Item 111 The Electron Transport Chain—Generating Proton
Motive Force 142
5.3 Using Heat to Destroy Microorganisms ATP Synthase—Harvesting the Proton Motive Force to
and Viruses 111 Synthesize ATP 145
Moist Heat 112 ATP Yield of Aerobic Respiration in Prokaryotes 146
Dry Heat 114
6.5 Fermentation 147
5.4 Using Other Physical Methods to Remove
or Destroy Microbes 114 6.6 Catabolism of Organic Compounds Other Than
Filtration 114 Glucose 149
Radiation 115 Polysaccharides and Disaccharides 150
High Pressure 116 Lipids 150
Proteins 150
5.5 Using Chemicals to Destroy Microorganisms
and Viruses 116 6.7 Chemolithotrophs 150
Potency of Germicidal Chemical Formulations 116 6.8 Photosynthesis 151
Selecting the Appropriate Germicidal Chemical 116 Capturing Radiant Energy 151
Classes of Germicidal Chemicals 117 Converting Radiant Energy into Chemical Energy 152
5.6 Preservation of Perishable Products 121 6.9 Carbon Fixation 154
Chemical Preservatives 121 Calvin Cycle 154
Low-Temperature Storage 121
Reducing the Available Water 121
xxviii Contents

6.10 Anabolic Pathways—Synthesizing Subunits from

Precursor Molecules 155
8 Bacterial Genetics 188
Lipid Synthesis 156
A Glimpse of History 188
Amino Acid Synthesis 156
Key Terms 188
Nucleotide Synthesis 157
8.1 Genetic Change
PERSPECTIVE 6.1: Mining with Microbes 151 in Bacteria 189
FUTURE CHALLENGES: Fueling the Future 157
MUTATION AS A MECHANISM
SUMMARY 158
OF GENETIC CHANGE
REVIEW QUESTIONS 159
8.2 Spontaneous Mutations 190
Base Substitution 190
7 The Blueprint of Life, from DNA Deletion or Addition of Nucleotides 191
to Protein 161 Transposons (Jumping Genes) 192
8.3 Induced Mutations 192
A Glimpse of History 161
Chemical Mutagens 192
Key Terms 161
Transposition 194
7.1 Overview 162 Radiation 194
Characteristics of DNA 162
8.4 Repair of Damaged DNA 194
Characteristics of RNA 163
Repair of Errors in Nucleotide Incorporation 195
Regulating Gene Expression 164
Repair of Modified Nucleobases in DNA 196
7.2 DNA Replication 165 Repair of Thymine Dimers 196
Initiation of DNA Replication 166 SOS Repair 196
The Process of DNA Replication 166
8.5 Mutant Selection 197
7.3 Gene Expression in Bacteria 168 Direct Selection 197
Transcription 168 Indirect Selection 197
Translation 170 Screening for Possible Carcinogens 199
7.4 Differences Between Eukaryotic and Prokaryotic HORIZONTAL GENE TRANSFER AS A MECHANISM
Gene Expression 176 OF GENETIC CHANGE
7.5 Sensing and Responding to Environmental 8.6 DNA-Mediated Transformation 202
Fluctuations 177 Competence 203
Signal Transduction 177 The Process of Transformation 204
Natural Selection 178
8.7 Transduction 204
7.6 Bacterial Gene Regulation 179
Mechanisms to Control Transcription 179 8.8 Conjugation 206
The lac Operon as a Model 181 Plasmid Transfer 206
Chromosome Transfer 207
7.7 Eukaryotic Gene Regulation 182 F Donors 208
7.8 Genomics 184 8.9 The Mobile Gene Pool 208
Analyzing a Prokaryotic DNA Sequence 184 Plasmids 208
Metagenomics 184 Transposons 209
Genomic Islands 211
PERSPECTIVE 7.1: RNA: The First Macromolecule? 177
FUTURE CHALLENGES: Gems in the Genomes? 185 PERSPECTIVE 8.1: The Biological Function of DNA: A Discovery
Ahead of Its Time 203
SUMMARY 185
PERSPECTIVE 8.2: Bacteria Can Conjugate with Plants: A Natural
REVIEW QUESTIONS 186 Case of Genetic Engineering 210
FUTURE CHALLENGES: Hunting for Magic Bullets 212

SUMMARY 212
REVIEW QUESTIONS 213
 Contents xxix

10.2 Using Phenotypic Characteristics to Identify

9 Biotechnology and Recombinant DNA 215
Prokaryotes 241
Microscopic Morphology 241
A Glimpse of History 215
Culture Characteristics 242
Key Terms 215
Metabolic Capabilities 243
9.1 Fundamental Tools Used Serology 245
in Biotechnology 216 Fatty Acid Analysis (FAME) 245
Restriction Enzymes 216
10.3 Using Genotypic Characteristics to Identify
Gel Electrophoresis 217
Prokaryotes 245
9.2 Applications of Genetic Engineering 218 Detecting Specific Nucleotide Sequences 245
Genetically Engineered Bacteria 218 Sequencing Ribosomal RNA Genes 246
Genetically Engineered Eukaryotes 220
10.4 Characterizing Strain Differences 247
9.3 Techniques Used in Genetic Engineering 221 Biochemical Typing 247
Obtaining DNA 222 Serological Typing 247
Generating a Recombinant DNA Molecule 222 Molecular Typing 247
Obtaining Cells That Harbor Recombinant DNA Phage Typing 248
Molecules 223 Antibiograms 248
9.4 Concerns Regarding Genetic Engineering 10.5 Classifying Prokaryotes 249
and Other DNA Technologies 224 16S rDNA Sequence Analysis 250
9.5 DNA Sequencing 224 DNA Hybridization 252
Techniques Used in DNA Sequencing 225 DNA Base Ratio (G+C Content) 252
Phenotypic Methods 252
9.6 Polymerase Chain Reaction (PCR) 227
Techniques Used in PCR 228 PERSPECTIVE 10.1: Tracing the Source of an Outbreak of Foodborne
Disease 251
9.7 Probe Technologies 232 FUTURE CHALLENGES: Tangled Branches in the Phylogenetic Tree 252
Colony Blotting 232
SUMMARY 253
Fluorescence in situ Hybridization (FISH) 232
REVIEW QUESTIONS 253
DNA Microarrays 233

PERSPECTIVE 9.1: Science Takes the Witness Stand 229

SUMMARY 234
11 The Diversity of Prokaryotic
Organisms 255
REVIEW QUESTIONS 235
A Glimpse of History 255
Key Terms 255
PART I I
The Microbial World METABOLIC DIVERSITY
11.1 Anaerobic
10 Identification and Classification of
Chemotrophs 256
Prokaryotic Organisms 237
Anaerobic Chemolithotrophs 256
Anaerobic Chemoorganotrophs—Anaerobic
A Glimpse of History 237
Respiration 258
Key Terms 237
Anaerobic Chemoorganotrophs—Fermentation 258
10.1 Principles of
11.2 Anoxygenic Phototrophs 259
Taxonomy 238
The Purple Bacteria 259
Strategies Used to Identify
The Green Bacteria 260
Prokaryotes 238
Other Anoxygenic Phototrophs 261
Strategies Used to Classify Prokaryotes 238
Nomenclature 241 11.3 Oxygenic Phototrophs 261
The Cyanobacteria 261
xxx Contents

11.4 Aerobic Chemolithotrophs 262 12.3 Protozoa 291

The Sulfur-Oxidizing Bacteria 262 Types of Protozoa 292
The Nitrifiers 263 Structure of Protozoa 293
The Hydrogen-Oxidizing Bacteria 264 Protozoan Habitats 293
Protozoan Reproduction 293
11.5 Aerobic Chemoorganotrophs 264
Medical Importance of Protozoa 294
Obligate Aerobes 264
Facultative Anaerobes 265 12.4 Slime Molds and Water Molds 294
Slime Molds 294
ECOPHYSIOLOGICAL DIVERSITY Water Molds 294

11.6 Thriving in Terrestrial Environments 266 12.5 Multicellular Parasites: Helminths 295
Bacteria That Form a Resting Stage 266 Roundworms 296
Bacteria That Associate with Plants 268 Tapeworms 298
Flukes 298
11.7 Thriving in Aquatic Environments 269
Sheathed Bacteria 269 12.6 Arthropods 298
Prosthecate Bacteria 270 Mosquitoes 299
Bacteria That Derive Nutrients from Other Fleas 300
Organisms 271 Lice 300
Bacteria That Move by Unusual Mechanisms 272 Ticks 300
Bacteria That Form Storage Granules 273 Mites 300

11.8 Animals as Habitats 273 PERSPECTIVE 12.1: What Causes River Blindness? 296
Bacteria That Inhabit the Skin 273 FUTURE CHALLENGES: The Continued Fight to Eradicate
Bacteria That Inhabit Mucous Membranes 275 Malaria 301
Obligate Intracellular Parasites 276 SUMMARY 301
11.9 Archaea That Thrive in Extreme Conditions 278 REVIEW QUESTIONS 303
Extreme Halophiles 278
Extreme Thermophiles 278 13 Viruses, Viroids, and Prions 304

FUTURE CHALLENGES: Astrobiology: The Search for Life A Glimpse of History 304
on Other Planets 279
Key Terms 304
SUMMARY 280
13.1 General Characteristics
REVIEW QUESTIONS 281 of Viruses 305
Viral Architecture 306
12 The Eukaryotic Members of the Microbial Viral Taxonomy 308
World 282
13.2 Bacteriophages 310
A Glimpse of History 282 Lytic Phage Infections: T4 Phage as a Model 310
Key Terms 282 Temperate Phage Infections: Lambda Phage as a
Model 312
12.1 Fungi 283 Consequences of Lysogeny 313
Types of Fungi 284 Filamentous Phages: M13 Phage as a Model 313
Structure of Fungi 284
13.3 The Roles of Bacteriophages in Horizontal Gene
Fungal Habitats 285
Transfer 314
Symbiotic Relationships of Fungi 286
Generalized Transduction 314
Reproduction in Fungi 286
Specialized Transduction 314
Economic Importance of Fungi 288
Medical Importance of Fungi 288 13.4 Bacterial Defenses Against Phages 315
Preventing Phage Attachment 315
12.2 Algae 289
Restriction-Modification Systems 315
Types of Algae 289
CRISPR System 316
Structure of Algae 289
Algal Habitats 290 13.5 Methods Used to Study
Algal Reproduction 291 Bacteriophages 317
Medical Importance of Algae 291
 Contents xxxi

13.6 Animal Virus Replication 317 14.5 Pattern Recognition Receptors (PRRs) 342
Attachment 317 Toll-Like Receptors (TLRs) 342
Penetration and Uncoating 318 NOD-Like Receptors (NLRs) 342
Synthesis of Viral Proteins and Replication RIG-Like Receptors (RLRs) 343
of the Genome 319
14.6 The Complement System 344
Assembly 320
Complement System Activation 345
Release 320
Effector Functions of the Complement System 345
13.7 Categories of Animal Virus Infections 321 Regulation of the Complement System 346
Acute Infections 322
Persistent Infections 322 14.7 Phagocytosis 346
The Process of Phagocytosis 346
13.8 Viruses and Human Tumors 323 Specialized Attributes of Macrophages 348
13.9 Cultivating and Quantitating Animal Viruses 324 Specialized Attributes of Neutrophils 348
Cultivating Animal Viruses 325 14.8 The Inflammatory Response 348
Quantitating Animal Viruses 326 Factors That Trigger an Inflammatory Response 348
13.10 Plant Viruses 326 The Inflammatory Process 349
Damaging Effects of Inflammation 350
13.11 Other Infectious Agents: Viroids and Prions 328 Cell Death and the Inflammatory Process 350
Viroids 328
Prions 328 14.9 Fever 351

PERSPECTIVE 13.1: Microbe Mimicker 306 PERSPECTIVE 14.1: For Schistosoma, the Inflammatory Response
Delivers 350
FUTURE CHALLENGES: Gene Therapy: Turning a Corner? 330
SUMMARY 351
SUMMARY 330
REVIEW QUESTIONS 352
REVIEW QUESTIONS 332

15 The Adaptive Immune Response 354

PART I I I A Glimpse of History 354

Microorganisms and Humans Key Terms 354

15.1 Strategy of the Adaptive

14 The Innate Immune Response 334
Immune Response 355
Overview of Humoral
A Glimpse of History 334
Immunity 355
Key Terms 334
Overview of Cell-Mediated Immunity 356
14.1 Overview of the Innate
15.2 Anatomy of the Lymphatic System 357
Defenses 335
Lymphatic Vessels 358
14.2 First-Line Defenses 336 Secondary Lymphoid Organs 358
Physical Barriers 336 Primary Lymphoid Organs 358
Antimicrobial Substances 337
15.3 The Nature of Antigens 359
Normal Microbiota (Flora) 337
14.3 The Cells of the Immune System 338 15.4 The Nature of Antibodies 359
Granulocytes 339 Structure and Properties of Antibodies 360
Mononuclear Phagocytes 340 Protective Outcomes of Antibody-Antigen Binding 360
Dendritic Cells 340 Immunoglobulin Classes 361
Lymphocytes 340 15.5 Clonal Selection and Expansion
of Lymphocytes 363
14.4 Cell Communication 341
Surface Receptors 341 15.6 B Lymphocytes and the Antibody Response 365
Cytokines 341 B-Cell Activation 365
Adhesion Molecules 342 Characteristics of the Primary Response 365
xxxii Contents

Characteristics of the Secondary Response 367 16.7 Avoiding the Host Defenses 388
The Response to T-Independent Antigens 367 Hiding Within a Host Cell 388
Avoiding Killing by Complement System Proteins 389
15.7 T Lymphocytes: Antigen Recognition
and Response 368 Avoiding Destruction by Phagocytes 389
General Characteristics of T Cells 368 Avoiding Antibodies 390
Activation of T Cells 370 16.8 Damage to the Host 391
Effector Functions of TC (CD8) Cells 372 Exotoxins 391
Effector Functions of TH (CD4) Cells 372 Endotoxin and Other Bacterial Cell Wall
Subsets of Dendritic Cells and T Cells 373 Components 394
Damaging Effects of the Immune Response 395
15.8 Natural Killer (NK) Cells 374
16.9 Mechanisms of Viral Pathogenesis 395
15.9 Lymphocyte Development 374
Binding to Host Cells and Invasion 395
Generation of Diversity 375
Avoiding Immune Responses 395
Negative Selection of Self-Reactive B Cells 377
Positive and Negative Selection of Self-Reactive 16.10 Mechanisms of Eukaryotic Pathogenesis 396
T Cells 377 Fungi 396
Protozoa and Helminths 397
PERSPECTIVE 15.1: What Flavors Are Your Major Histocompatibility
Complex Molecules? 370 FUTURE CHALLENGES: The Potential of Probiotics 397
SUMMARY 377 SUMMARY 398
REVIEW QUESTIONS 378 REVIEW QUESTIONS 399

16 Host-Microbe Interactions 380

17 Immunologic Disorders 401

A Glimpse of History 401

A Glimpse of History 380 Key Terms 401
Key Terms 380
17.1 Type I Hypersensitivities:
MICROBES, HEALTH, Immediate
AND DISEASE IgE-Mediated 402
16.1 The Anatomical Barriers Localized Allergic Reactions 403
as Ecosystems 381 Systemic Anaphylaxis 404
Treatments to Prevent Allergic Reactions 404
16.2 The Normal Microbiota 381
The Protective Role of the Normal Microbiota 382 17.2 Type II Hypersensitivities: Cytotoxic 405
The Dynamic Nature of the Normal Microbiota 382 Transfusion Reactions 405
Hemolytic Disease of the Newborn 406
16.3 Principles of Infectious Disease 382
Pathogenicity 382 17.3 Type III Hypersensitivities: Immune
Characteristics of Infectious Disease 383 Complex–Mediated 407

16.4 Establishing the Cause of Infectious Disease 384 17.4 Type IV Hypersensitivities: Delayed-
Type Cell-Mediated 409
Koch’s Postulates 384
Tuberculin Skin Test 409
Molecular Koch’s Postulates 385
Delayed-Type Hypersensitivity in Infectious
Diseases 409
MECHANISMS OF PATHOGENESIS
Contact Hypersensitivities 409
16.5 Establishing Infection 386
17.5 Rejection of Transplanted Tissues 411
Adherence 386
Colonization 386 17.6 Autoimmune Disease 412
Delivering Effector Proteins to Host Cells 386 The Spectrum of Autoimmune Diseases 412
Treatment of Autoimmune Diseases 413
16.6 Invasion—Breaching the Anatomical Barriers 387
Penetrating the Skin 387 17.7 Immunodeficiency Disorders 414
Penetrating Mucous Membranes 387 Primary Immunodeficiencies 414
Secondary Immunodeficiencies 415
 Contents xxxiii

PERSPECTIVE 17.1: The Fetus as an Allograft 412 Reservoirs of Infection 438

FUTURE CHALLENGES: New Approaches to Correcting Immunologic Portals of Exit and Portals of Entry 439
Disorders 416 Disease Transmission 440
SUMMARY 416 Pathogen Factors That Influence the Epidemiology
REVIEW QUESTIONS 417 of Disease 442
Host Factors That Influence the Epidemiology
18 Applications of Immune Responses 419 of Disease 443
19.2 Epidemiological Studies 444
A Glimpse of History 419 Descriptive Studies 444
Key Terms 419 Analytical Studies 445
Experimental Studies 445
IMMUNIZATION
19.3 Infectious Disease Surveillance 446
18.1 Principles of
Immunization 420 National Disease Surveillance Network 446
Active Immunity 420 Worldwide Disease Surveillance 447
Passive Immunity 420 Reduction and Eradication of Disease 447

18.2 Vaccines and Immunization Procedures 421 19.4 Emerging Infectious Diseases 448
Attenuated Vaccines 423 19.5 Healthcare-Associated Infections 449
Inactivated Vaccines 423 Reservoirs of Infectious Agents in Healthcare
An Example of Vaccination Strategy—The Campaign Settings 450
to Eliminate Poliomyelitis 424 Transmission of Infectious Agents in Healthcare
The Importance of Childhood Immunizations 424 Settings 451
Current Progress in Immunization 425 Preventing Healthcare-Associated Infections 452

IMMUNOLOGICAL TESTING PERSPECTIVE 19.1: Standard Precautions—Protecting Patients

and Healthcare Personnel 452
18.3 Principles of Immunological Testing 426
FUTURE CHALLENGES: Maintaining Vigilance Against
Obtaining Antibodies 426 Bioterrorism 453
Quantifying Antigen-Antibody Reactions 427
SUMMARY 454
18.4 Observing Antigen-Antibody Aggregates 427 REVIEW QUESTIONS 455
Precipitation Reactions 427
Agglutination Reactions 430
18.5 Using Labeled Antibodies to Detect Antigen-
20 Antimicrobial Medications 457
Antibody Interactions 430
A Glimpse of History 457
Basic Principles 430
Key Terms 457
Fluorescent Antibody (FA) Test 431
Enzyme-Linked Immunosorbent Assay (ELISA) 432 20.1 History and Development
Western Blotting 432 of Antimicrobial
Fluorescence-Activated Cell Sorter (FACS) 434 Drugs 458
Discovery of Antimicrobial Drugs 458
PERSPECTIVE 18.1: Monoclonal Antibodies 428 Discovery of Antibiotics 458
FUTURE CHALLENGES: Global Immunization 434 Development of New Generations of Drugs 458
SUMMARY 434 20.2 Features of Antimicrobial Drugs 458
REVIEW QUESTIONS 435 Selective Toxicity 459
Antimicrobial Action 459
19 Epidemiology 437 Spectrum of Activity 459
Effects of Combinations 460
A Glimpse of History 437
Tissue Distribution, Metabolism, and Excretion
Key Terms 437
of the Drug 460
19.1 Principles of Adverse Effects 460
Epidemiology 438 Resistance to Antimicrobials 460
Rate of Disease in a
Population 438
xxxiv Contents

20.3 Mechanisms of Action of Antibacterial Drugs 461

Antibacterial Medications That Inhibit Cell Wall
21 Respiratory System Infections 483
Synthesis 463
A Glimpse of History 483
Antibacterial Medications That Inhibit Protein
Key Terms 483
Synthesis 465
Antibacterial Medications That Inhibit Nucleic Acid 21.1 Anatomy, Physiology,
Synthesis 466 and Ecology 484
Antibacterial Medications That Interfere with Metabolic The Upper Respiratory
Pathways 467 Tract 484
Antibacterial Medications That Interfere with Cell The Lower Respiratory Tract 486
Membrane Integrity 467
Antibacterial Medications Effective Against INFECTIONS OF THE UPPER RESPIRATORY SYSTEM
Mycobacterium Species 467 21.2 Bacterial Infections of the Upper Respiratory
20.4 Determining the Susceptibility of a Bacterial System 486
Strain to an Antimicrobial Drug 468 Streptococcal Pharyngitis (“Strep Throat”) 487
Minimum Inhibitory and Bactericidal Concentrations Post-Streptococcal Sequelae 489
(MIC and MBC) 468 Diphtheria 490
Conventional Disc Diffusion Method 468 Pinkeye, Earache, and Sinus Infections 492
Commercial Modifications of Antimicrobial
21.3 Viral Infections of the Upper Respiratory
Susceptibility Testing 468
System 494
20.5 Resistance to Antimicrobial Drugs 471 The Common Cold 494
Mechanisms of Acquired Resistance 471 Adenoviral Respiratory Tract Infections 495
Acquisition of Resistance 472
Examples of Emerging Antimicrobial Resistance 472 INFECTIONS OF THE LOWER RESPIRATORY SYSTEM
Slowing the Emergence and Spread of Antimicrobial
Resistance 473 21.4 Bacterial Infections of the Lower Respiratory
System 496
20.6 Mechanisms of Action of Antiviral Drugs 474 Pneumococcal Pneumonia 496
Prevent Viral Entry 474 Klebsiella Pneumonia 498
Interfere with Viral Uncoating 475 Mycoplasmal Pneumonia (“Walking Pneumonia”) 500
Interfere with Nucleic Acid Synthesis 475 Pertussis (“Whooping Cough”) 500
Prevent Genome Integration 476 Tuberculosis (“TB”) 502
Prevent Assembly and Release of Viral Particles 476 Legionnaires’ Disease 506
20.7 Mechanisms of Action of Antifungal 21.5 Viral Infections of the Lower Respiratory
Drugs 476 System 507
Interfere with Plasma Membrane Synthesis Influenza (“Flu”) 508
and Function 476
Respiratory Syncytial Virus Infections 511
Interfere with Cell Wall Synthesis 477
Hantavirus Pulmonary Syndrome 512
Interfere with Cell Division 477
Interfere with Nucleic Acid Synthesis 477 21.6 Fungal Infections of the Lung 514
Coccidioidomycosis (“Valley Fever”) 514
20.8 Mechanisms of Action of Antiprotozoan
Histoplasmosis (“Spelunker’s Disease”) 515
and Antihelminthic Drugs 478
PERSPECTIVE 21.1: Terror by Mail: Inhalation Anthrax 497
PERSPECTIVE 20.1: Measuring the Concentration
PERSPECTIVE 21.2: What to Do About Bird Flu 509
of an Antimicrobial Drug in Blood or Other Body
Fluids 470 FUTURE CHALLENGES: Global Preparedness Versus Emerging
Respiratory Viruses 518
FUTURE CHALLENGES: War with the Superbugs 479
SUMMARY 518
SUMMARY 479
REVIEW QUESTIONS 519
REVIEW QUESTIONS 481
 Contents xxxv

23.4 Bacterial Infections of Bite Wounds 561

22 Skin Infections 521
Human Bites 561
Pasteurella multocida Bite Wound Infections 562
A Glimpse of History 521
Key Terms 521
Bartonellosis (“Cat Scratch Disease”) 563
Streptobacillary Rat Bite Fever 564
22.1 Anatomy, Physiology,
and Ecology 522 23.5 Fungal Wound Infections 565
Sporotrichosis (“Rose Gardener’s Disease”) 565
22.2 Bacterial Skin
Diseases 523 CASE PRESENTATION 560
Acne Vulgaris 523 PERSPECTIVE 23.1: Infection Caused by a Human “Bite“ 562
Hair Follicle Infections 524 FUTURE CHALLENGES: Staying Ahead in the Race with
Staphylococcal Scalded Skin Syndrome 527 Staphylococcus aureus 568

Streptococcal Impetigo 528 SUMMARY 568

Rocky Mountain Spotted Fever 529 REVIEW QUESTIONS 569
Lyme Disease 531
22.3 Skin Diseases Caused by Viruses 534
24 Digestive System Infections 571
Varicella (Chickenpox) 534
A Glimpse of History 571
Rubeola (Measles) 537
Key Terms 571
Rubella (German Measles) 539
Other Viral Rashes of Childhood 541 24.1 Anatomy, Physiology,
Warts 542 and Ecology 572
The Upper Digestive
22.4 Skin Diseases Caused by Fungi 543
System 573
Superficial Cutaneous Mycoses 543
The Lower Digestive System 574
Other Fungal Diseases 544
UPPER DIGESTIVE SYSTEM INFECTIONS
CASE PRESENTATION 540
PERSPECTIVE 22.1: The Ghost of Smallpox: An Evil Shade 536 24.2 Bacterial Diseases of the Upper Digestive
FUTURE CHALLENGES: The Ecology of Lyme Disease 546 System 575
Dental Caries (Tooth Decay) 575
SUMMARY 546 Periodontal Disease 577
REVIEW QUESTIONS 547 Acute Necrotizing Ulcerative Gingivitis 578
Helicobacter pylori Gastritis 578
23 Wound Infections 548 24.3 Viral Diseases of the Upper Digestive System 581
Herpes Simplex (Cold Sores) 581
A Glimpse of History 548 Mumps 583
Key Terms 548
LOWER DIGESTIVE SYSTEM INFECTIONS
23.1 Anatomy, Physiology,
24.4 Bacterial Diseases of the Lower Digestive
and Ecology 549
System 584
Wound Abscesses 550
General Characteristics 585
Anaerobic Wounds 550
Cholera 586
23.2 Common Bacterial Infections of Wounds 551 Shigellosis 588
Staphylococcal Wound Infections 551 Escherichia coli Gastroenteritis 589
Group A Streptococcal “Flesh-Eating Disease” 553 Salmonella Gastroenteritis 592
Pseudomonas aeruginosa Infections 554 Typhoid and Paratyphoid Fevers 592
Campylobacteriosis 593
23.3 Diseases Due to Anaerobic Bacterial Wound
Infections 555 Clostridium difficile–Associated Disease (CDAD) 594
Tetanus (“Lockjaw”) 555 24.5 Viral Diseases of the Lower Digestive System—
Clostridial Myonecrosis (“Gas Gangrene”) 558 Intestinal Tract 595
Actinomycosis (“Lumpy Jaw”) 559 Rotaviral Gastroenteritis 595
Norovirus Gastroenteritis 596
xxxvi Contents

24.6 Viral Diseases of the Lower Digestive System— CASE PRESENTATION 614
Liver 597 PERSPECTIVE 25.1: The Death of Syphilis? 628
Hepatitis A 597 FUTURE CHALLENGES: Getting Control of Sexually Transmitted
Hepatitis B 599 Infections 636
Hepatitis C 601 SUMMARY 638
24.7 Protozoan Diseases of the Lower Digestive REVIEW QUESTIONS 639
System 602
Giardiasis 602 26 Nervous System Infections 641
Cryptosporidiosis (“Crypto”) 603
Cyclosporiasis 604 A Glimpse of History 641
Amebiasis 605 Key Terms 641

CASE PRESENTATION 581 26.1 Anatomy, Physiology,

PERSPECTIVE 24.1: Ecology of Cholera 586 and Ecology 641
FUTURE CHALLENGES: Defeating Digestive Tract Diseases 608 26.2 Bacterial Diseases
SUMMARY 608 of the Nervous System 644
REVIEW QUESTIONS 609 Pneumococcal Meningitis 644
Meningococcal Meningitis 645
Haemophilus influenzae Meningitis 646
25 Genitourinary Tract Infections 611 Neonatal Meningitis 648
Listeriosis 648
A Glimpse of History 611 Hansen’s Disease (Leprosy) 650
Key Terms 611 Botulism 652

25.1 Anatomy, Physiology, 26.3 Viral Diseases of the Nervous System 654
and Ecology 612 Viral Meningitis 654
The Urinary System 612 Viral Encephalitis 655
The Genital System 612 Poliomyelitis 656
Rabies 658
25.2 Urinary Tract Infections 613
Bacterial Cystitis (“Bladder Infection”) 613 26.4 Fungal Diseases of the Nervous System 660
Leptospirosis 615 Cryptococcal Meningoencephalitis 660
25.3 Genital System Diseases 617 26.5 Protozoan Diseases of the Nervous System 661
Bacterial Vaginosis (BV) 617 African Sleeping Sickness 662
Vulvovaginal Candidiasis (VVC) 618 Primary Amebic Meningoencephalitis (PAM) 663
Staphylococcal Toxic Shock Syndrome 619
26.6 Diseases Caused by Prions 664
25.4 Sexually Transmitted Infections: Scope Transmissible Spongiform Encephalopathy
of the Problem 620 in Humans 664
25.5 Bacterial STIs 622 CASE PRESENTATION 647
Gonorrhea 622 PERSPECTIVE 26.1: Rabies Survivors! 659
Chlamydial Infections 625 FUTURE CHALLENGES: Eradicate Polio: Then What? 667
Syphilis 626
SUMMARY 667
Chancroid 629
REVIEW QUESTIONS 668
25.6 Viral STIs 630
Genital Herpes 630 27 Blood and Lymphatic Infections 670
Papillomavirus STIs: Genital Warts and Cervical
Cancer 631 A Glimpse of History 670
HIV/AIDS 633 Key Terms 670

25.7 Protozoal STIs 635 27.1 Anatomy, Physiology,

Trichomoniasis (“Trich”) 635 and Ecology 671
The Heart 671
Blood Vessels 671
 Contents xxxvii

Lymphatics (Lymphatic Vessels) 672 Cytomegalovirus Disease 711

Spleen 672 Mycobacterial Diseases 713
27.2 Bacterial Diseases of the Blood Vascular CASE PRESENTATION 706
System 672 PERSPECTIVE 28.1: Origin of AIDS-Causing Viruses 698
Subacute Bacterial Endocarditis (SBE) 673 FUTURE CHALLENGES: AIDS and Poverty 714
Sepsis and Septic Shock 674
SUMMARY 716
27.3 Bacterial Diseases of the Lymph Nodes REVIEW QUESTIONS 717
and Spleen 675
Tularemia (“Rabbit Fever” or “Deer Fly Fever”) 675
Brucellosis (“Undulant Fever” or “Bang’s Disease”) 676 P AR T V
Plague (“Black Death”) 678
Applied Microbiology
27.4 Viral Diseases of the Lymphatic and Blood
Vascular Systems 680
Infectious Mononucleosis (“Mono” or “Kissing
29 Microbial Ecology 718
Disease”) 680
A Glimpse of History 718
Yellow Fever 682
Key Terms 718
Dengue Fever 683
Chikungunya 685 29.1 Principles of Microbial
Emerging Hemorrhagic Fevers 685 Ecology 719
Nutrient Acquisition 719
27.5 Protozoan Diseases 686
Microbes in Low-Nutrient Environments 720
Malaria 686
Microbial Competition and Antagonism 720
CASE PRESENTATION 684 Microorganisms and Environmental Changes 720
PERSPECTIVE 27.1: Arteriosclerosis: The Infection Hypothesis 671 Microbial Communities 721
PERSPECTIVE 27.2: Walter Reed and Yellow Fever 682 Studying Microbial Ecology 721
FUTURE CHALLENGES: Rethinking Malaria Control 691 29.2 Aquatic Habitats 722
SUMMARY 691 Marine Environments 722
REVIEW QUESTIONS 692 Freshwater Environments 723
Specialized Aquatic Environments 723
28 HIV Disease and Complications 29.3 Terrestrial Habitats 724
of Immunodeficiency 694 Characteristics of Soil 724
Microorganisms in Soil 724
A Glimpse of History 694 The Rhizosphere 724
Key Terms 694
29.4 Biogeochemical Cycling and Energy Flow 725
AIDS Carbon Cycle 725
28.1 Human Immunodeficiency Nitrogen Cycle 726
Virus (HIV) Infection and Sulfur Cycle 728
AIDS 695 Phosphorus Cycle and Other Cycles 728
HIV Disease 696 Energy Sources for Ecosystems 728
HIV Vaccine Prospects 704 29.5 Mutualistic Relationships Between
Microorganisms and Eukaryotes 729
COMPLICATIONS OF ACQUIRED IMMUNODEFICIENCIES
Mycorrhizas 730
28.2 Malignant Tumors 707 Symbiotic Nitrogen-Fixers and Plants 730
Kaposi’s Sarcoma 707 Microorganisms and Herbivores 732
B-Lymphocyte Tumors 707 SUMMARY 732
Cervical and Anal Carcinoma 708 REVIEW QUESTIONS 733
28.3 Infectious Diseases 708
Pneumocystis pneumonia (PCP) 708
Toxoplasmosis 709
xxxviii Contents

30 Environmental Microbiology: Treatment of 31.2 Microorganisms in Food and Beverage

Production 750
Water, Wastes, and Polluted Habitats 735
Lactic Acid Fermentations by the Lactic Acid
Bacteria 751
A Glimpse of History 735
Alcoholic Fermentations by Yeast 753
Key Terms 735
Changes Due to Mold Growth 755
30.1 Microbiology
31.3 Food Spoilage 756
of Wastewater
Treatment 736 Common Spoilage Bacteria 756
Biochemical Oxygen Demand (BOD) 736 Common Spoilage Fungi 756
Municipal Wastewater Treatment Methods 736 31.4 Foodborne Illness 756
Individual Wastewater Treatment Systems 740 Foodborne Intoxication 756
30.2 Drinking Water Treatment and Testing 740 Foodborne Infection 758
Water Treatment Processes 741 31.5 Food Preservation 759
Water Testing 742
PERSPECTIVE 31.1: Botox for Beauty and Pain Relief 758
30.3 Microbiology of Solid Waste Treatment 743
FUTURE CHALLENGES: Using Microorganisms to Nourish the
Sanitary Landfills for Solid Waste Disposal 743 World 760
Municipal and Backyard Composting—Alternative
SUMMARY 760
to Landfills 743
REVIEW QUESTIONS 761
30.4 Microbiology of Bioremediation 744
Pollutants 744 APPENDIX I Microbial Mathematics A–1
Means of Bioremediation 745
APPENDIX II Pronunciation Key for Bacterial, Fungal,
PERSPECTIVE 30.1: What a Gas! 739 Protozoan, and Viral Names A–2

FUTURE CHALLENGES: Better Identification of Pathogens in Water APPENDIX III Metabolic Pathways A–4
and Wastes 746
APPENDIX IV Answers to Multiple Choice Questions A–8
SUMMARY 746
GLOSSARY G–1
REVIEW QUESTIONS 747
CREDITS C–1
INDEX I–1
31 Food Microbiology 748

A Glimpse of History 748

Key Terms 748

31.1 Factors Influencing

the Growth of
Microorganisms in
Foods 749
Intrinsic Factors 749
Extrinsic Factors 750
1 Humans and
the Microbial World
KEY TERMS
Domain The highest level in
classification above kingdom.
There are three domains: Bacteria,
Archaea, and Eucarya.
Prokaryote Single-celled organism
consisting of a prokaryotic cell;
members of the domains Bacteria
and Archaea are prokaryotes.
Eukaryote Organism composed Prokaryotic Cell Cell type
of one or more eukaryotic cells; characterized by the lack of a
members of the domain Eucarya are membrane-bound nucleus.
eukaryotes. Viroid A non-living infectious
Eukaryotic Cell Cell type agent consisting only of RNA.
characterized by a membrane-bound Virus A non-living infectious agent
nucleus. consisting of nucleic acid surrounded
Obligate Intracellular Parasite by a protein coat.
An organism or virus that multiplies
only inside living cells.
Prion A non-living infectious agent
consisting only of protein.

Drawings that van Leeuwenhoek made in 1683 of the shapes of microorganisms mold. Hooke also described how to make the kind of microscope that van
he saw through his single lens microscope. He also observed organism B moving Leeuwenhoek constructed almost 10 years later. Both men deserve equal
from position C to D. credit for revealing the world of microbes—the organisms you are about
to study.

A Glimpse of History

M
icroorganisms are the foundation for all life on earth.
Microbiology as a science was born in 1674 when Antony van
Leeuwenhoek (1632–1723), an inquisitive Dutch drapery merchant, They have existed on this planet for about 3.5 billion
peered at a drop of lake water through a glass lens he had carefully years, and over this time, plants, animals, and modern
made. For several centuries it was known that curved glass would mag- microorganisms evolved from them. Microorganisms may be
nify objects, but the skilled hands of a craftsman and his questioning invisible to the naked eye, but our life depends on their activities.
mind uncovered an entirely new part of our world. What he observed
through his simple magnifying glass was undoubtedly one of the most
startling and amazing sights that humans have ever beheld—the world of
microbes. As van Leeuwenhoek wrote in a letter to the Royal Society of
London, he saw
1.1 ■ The Dispute Over
Very many little animalcules, whereof some were roundish,
Spontaneous Generation
while others a bit bigger consisted of an oval. On these last, I
Learning Outcome
saw two little legs near the head, and two little fins at the hind
most end of the body. Others were somewhat longer than an oval, 1. Describe the key experiments of scientists who disproved
and these were very slow a-moving, and few in number. These spontaneous generation.
animalcules had diverse colours, some being whitish and trans-
parent; others with green and very glittering little scales, others The discovery of microorganisms raised an intriguing question:
again were green in the middle, and before and behind white; “Where did these microscopic forms originate?” Some people
others yet were ashed grey. And the motion of most of these believed that worms and other forms of life arise from non-living
animalcules in the water was so swift, and so various, upwards, material in a process referred to as spontaneous generation. This
downwards, and round about, that ’twas wonderful to see. was challenged by an Italian biologist and physician, Francesco
Before van Leeuwenhoek observed bacteria, Robert Hooke, an Redi. In 1668, he used a simple experiment to show that worms
English microscopist saw another kind of microorganism. In 1665, he found on rotting meat originated from the eggs of flies, not from
described what he called a “microscopical mushroom.” His drawing was the decaying meat as proponents of spontaneous generation
so accurate that his specimen could later be identified as a common bread believed. In his experiment, Redi covered the meat with fine gauze

1
2 Chapter 1 Humans and the Microbial World

that prevented flies from depositing their eggs. When he did this, clever experiments. First, he demonstrated that air contains
no worms appeared. microorganisms. He did this by filtering air through a cotton plug,
Despite Redi’s work that explained the source of worms on trapping microorganisms. He then examined the trapped microor-
decaying meat, convincing proof that microorganisms did not ganisms with a microscope and found that many looked identical
arise from spontaneous generation took over 200 more years! One to those described by others who had been studying broths. When
reason for the delay was that various experiments in different Pasteur dropped the cotton plug into a sterilized broth, the broth
laboratories gave conflicting results. became cloudy.
Most importantly, Pasteur demonstrated that sterile broths in
Early Experiments specially constructed swan-necked flasks remained sterile even
when left open to air (figure 1.1). Microorganisms from the air
In 1749, John Needham, a scientist and Catholic priest, showed
settled in the bends and sides of these flasks, never reaching the
that flasks containing various broths (made by soaking hay,
broth. Only when the flasks were tipped would bacteria enter the
chicken, or other nutrient source in water) gave rise to microor-
broth and grow. These simple and elegant experiments ended the
ganisms even when the flasks were boiled and sealed with a cork.
arguments that unheated air or the broths themselves contained a
At that time, brief boiling was thought to kill all organisms, so this
“vital force” necessary for spontaneous generation.
suggested that microorganisms did indeed arise spontaneously.
In 1776, the animal physiologist and priest, Father Spallanzani,
obtained results that contradicted the experiments of Needham; no Experiments of Tyndall
bacteria appeared in Spallazani’s broths after boiling. His experi-
Although most scientists were convinced by Pasteur’s experi-
ments differed from Needham’s in two significant ways: Spallanzani
ments, some remained skeptical because they could not reproduce
boiled the broths for longer periods and he sealed the flasks by melt-
his results. An English physicist, John Tyndall, finally explained
ing their glass necks. Using these techniques, he repeatedly demon-
the conflicting data and, in turn, proved Pasteur correct. Tyndall
strated that broths remained sterile (free of microorganisms).
found that various types of broths required different boiling times
However, if the neck of the flask cracked, the broth rapidly became
to be sterilized. Some materials were sterilized by boiling for 5
cloudy due to growth of the organisms. Spallanzani concluded
minutes, whereas others, most notably broths made from hay, still
microorganisms had entered the broth with the air, and the corks
contained living microorganisms even after boiling for 5 hours!
used by many investigators did not keep them out.
Even when hay was merely present in the laboratory, broths that
Spallanzani’s experiments did not stop the controversy. Some
had previously been sterilized by boiling for 5 minutes were some-
people argued that the heating process destroyed a “vital force” nec-
times impossible to sterilize. What was going on? Tyndall finally
essary for spontaneous generation. The debate continued and further
realized that hay contained heat-resistant forms of microbial life.
experiments, with proper controls, were needed to settle the matter.
When hay was brought into the laboratory, dust particles must have
transferred these heat-resistant forms to the broths.
Experiments of Pasteur Tyndall concluded that some microorganisms exist in two
One giant in science who helped disprove spontaneous generation forms: a cell readily killed by boiling, and one that is heat resis-
was Louis Pasteur, the French chemist considered by many to be tant. In the same year (1876), a German botanist, Ferdinand Cohn,
the father of modern microbiology. In 1861, he did a series of discovered endospores, the heat-resistant forms of bacteria.

Air escapes from Microorganisms from

open end of flask. air settle in bend.

Years Hours/days

1 Broth sterilized— 2 Broth allowed 3 Broth stays sterile 4 Flask tilted so that 5 Bacteria multiply
air escapes. to cool slowly— indefinitely. the sterile broth comes in broth.
air enters. in contact with micro-
organisms from air.

FIGURE 1.1 Pasteur’s Experiment with the Swan-Necked Flask If the flask remains upright, no microbial growth occurs. If the flask is
tipped, the microorganisms trapped in the neck reach the sterile broth and grow.
? If the broth in Pasteur’s swan-necked flasks had contained endospores, what results would have been observed?
 Part I Life and Death of Microorganisms 3

Robert Koch then demonstrated that anthrax was caused by a Vital Activities of Microorganisms
spore-forming bacterium.  endospores, p. 67
The activities of microorganisms are required for the survival of not
The extreme heat resistance of endospores explains the differ-
just humans, but all other organisms on this planet. A few examples
ences between Pasteur’s results and those of other investigators.
readily prove this point. Nitrogen is an essential part of nucleic acids
Organisms that produce endospores are commonly found in the
and proteins. Because of this, all life-forms need a continual supply
soil and most likely were present in broths made from hay. Pasteur
of this element. A plentiful source is N2—the most common gas in
used only broths made with sugar or yeast extract, so his experi-
the atmosphere—yet neither plants nor animals can use this form.
ments probably did not have endospores. At the time, scientists
Instead, we depend on certain microbes that convert N2 into a form
did not appreciate the importance of the source of the broth, but in
other organisms can use. Without these microbes, life as we know
hindsight, the source was critical. This points out an important les-
it would not exist.
son for all scientists. In repeating an experiment, it is essential to
Humans and other animals all require oxygen gas (O2) to
reproduce all conditions as closely as possible. What may seem like
breathe. The supply of O2 in the atmosphere, however, would be
a trivial difference may be extremely important.
depleted in about 20 years were it not continually replenished.
It may seem surprising that spontaneous generation was dis-
Plants produce O2 during photosynthesis, but so do many photo-
proved only about a century and a half ago. How far the science
synthetic microorganisms.
of microbiology and all biological sciences have advanced since
Microorganisms are the only organisms that can degrade
that time! Figure 1.2 lists some of the more important advances
certain materials. For example, humans and other animals cannot
made over the years in the context of other historical events. It
digest cellulose—an important component of plants—but some
also highlights a period called the Golden Age of Microbiology—a
microorganisms can. Because microbes are able to degrade cel-
time during which the field of microbiology blossomed. Rather
lulose, leaves and fallen trees do not pile up in the environment.
than cover more history now, we will return to many of these
Cellulose is also degraded by billions of microorganisms in the
milestones in brief stories that open each chapter.
digestive tracts of cattle, sheep, deer, and other ruminants. The
MicroByte digestion products are used by the ruminants for energy, and
In 2001, someone mailed anthrax endospores to selected individuals without them the animals would starve. Microorganisms also
in the United States, an act of bioterrorism. play indispensable roles in degrading a wide variety of materials
in sewage and wastewater.  ruminants, p. 732
MicroAssessment 1.1
Pasteur and Tyndall finally disproved spontaneous generation about Applications of Microbiology
150 years ago. In addition to the crucial roles that microorganisms play in main-
1. Give two reasons why it took so long to disprove spontaneous taining all life, they also have made life more comfortable for
generation. humans over the centuries.
2. What experiment disproved the notion that a “vital force” in air
was responsible for spontaneous generation? Food Production
3. What conclusions could Tyndall reach on the properties of Egyptian bakers as early as 2100 b.c. used yeast to make bread. Today,
the agent that entered the broth from hay? + bakeries use essentially the same technology.  breadmaking, p. 755
The excavation of early tombs in Egypt revealed that by
1500 b.c., Egyptians used a complex procedure for fermenting cereal
grains to produce beer. Today, brewers use the same fundamental
1.2 ■ Microbiology: A Human techniques to make beer and other fermented drinks.  beer, p. 754
Perspective Virtually every population that raised milk-producing animals
such as cows and goats also developed procedures to ferment milk.
Learning Outcomes This allowed them to make foods such as yogurt, cheeses, and but-
2. Explain why life could not exist without microorganisms. termilk. Today, the bacteria added to some fermented milk products
3. Describe three applications of microbiology. are touted as probiotics, protecting against intestinal infections and
4. Describe the role of microbes in disease, including the triumphs, bowel cancer.  fermented milk products, p. 751  probiotics, p. 397
present and future challenges, and host-microbe interactions.
Biodegradation
Microorganisms have an enormous impact on all living things. Microorganisms degrade environmental pollutants. For example,
We could not survive without them, and they also make our bacteria can destroy polychlorinated biphenyls (PCBs), dichlo-
lives far more comfortable. At the same time, microorganisms rodiphenyltrichloroethane (DDT), trichloroethylene, (a toxic sol-
and other infectious agents, collectively called microbes, have vent used in dry cleaning), and other chemicals in contaminated soil
killed far more people than have ever been killed in war. and water. Bacteria also lessen the damage from oil spills. In some
Microbes have even been used as weapons in times of war, and cases, microorganisms are added to pollutants to hasten their decay,
continue today to be used as agents of bioterrorism. a process called bioremediation.  bioremediation, p. 744
4 Chapter 1 Humans and the Microbial World

• Egyptians ferment • Pilgrims establish • Robert Hooke publishes his

cereal grains to make Plymouth Colony: 1620 discovery of cells and sees
beer: 1500 B.C. the first microorganism: 1665
• Harvard College,
• Tutankhamen in first college in the • Edward Jenner introduces
Egypt: 1300 B.C. U.S., founded: vaccination for smallpox: 1796
1636
• Lewis and Clark explore
Historical the West: 1804–1806
Events
• War of 1812 with
England: 1812–1814
Milestones in
Microbiology

• First permanent English • American Revolution:

• Ferdinand Magellan’s settlement in America, 1775–1783
ships circle the globe: Jamestown: 1607
1519–1522
• Antony van Leeuwenhoek
observes bacteria: 1676
• Girolamo Fracastoro suggests
that invisible organisms may
cause disease: 1546 • Napoleon defeated
at Waterloo: 1815

• Louis Pasteur demonstrates

that yeast can degrade sugar
• J. Henle proposes
to ethanol and carbon dioxide: 1857
the germ theory
• Louis Pasteur of disease: 1840
refutes spontaneous
generation: 1861

• Mathias Schleiden and

• Darwin publishes • John Snow demonstrates • U.S.–Mexican War: Theodor Schwann
Origin of Species: the epidemic spread of 1846–1848 propose that all
1859 cholera through a organisms are composed
contaminated water • Ignaz Semmelweis demonstrates of cells:1838–1839
• American Civil supply: 1853–1854
War: 1861–1865 that childbed fever is a contagious
disease transmitted by doctors
• Louis Pasteur develops during childbirth: 1847–1850
pasteurization to destroy • Christian Gram describes
organisms in wine: 1864 the Gram stain: 1884
• Elie Metchnikoff* discovers
• Blacks given the • Robert Koch introduces phagocytic cells which
right to vote in U.S.: 1870 pure culture techniques in engulf bacteria: 1884
• U.S. acquires the laboratory: 1881 • Koch states Koch’s
Alaska from • Robert Koch* demonstrates postulates: 1884
Russia: 1867 that a bacterium causes
anthrax: 1876

• Joseph Lister publishes

the first work on
antiseptic surgery: 1867 • Koch discovers the cause
of tuberculosis: 1882
• Custer’s Last • Walter and Fanny Hesse
Stand: 1876 introduce agar-agar as a • Dmitri Iwanowski discovers that
solidifying gel for culture a filterable agent, a virus, causes
media: 1882 tobacco-mosaic disease: 1892

FIGURE 1.2 Some major milestones in microbiology—and their timeline in relation to other historical events. The asterisks indicate scientists
who won the Nobel Prize. The gold band indicates the Golden Age of Microbiology.
? What is the Golden Age of Microbiology?
 Part I Life and Death of Microorganisms 5

• F. Peyton Rous discovers that a virus

can cause cancer in chickens: 1911 • Frederick Griffith discovers
• Wright Brothers • Women given the genetic transformation in
make controlled right to vote in U.S.: 1919 bacteria: 1928
sustained flight:
• Worldwide influenza • Alexander Fleming* discovers
1903
epidemic: 1918 first antibiotic, penicillin: 1929
Historical • Prohibition of alcoholic
Events beverages repealed: 1933

Milestones in • World War II:

Microbiology 1939–1945
• World War I: • Alcoholic beverages • First talking motion
1914–1918 prohibited in U.S.: 1920 picture made: 1928
• Paul Ehrlich develops • Beadle* and Tatum* show that genes
the drug Salvarsan, • First woman elected
to Congress: 1916 direct the synthesis of proteins: 1941
the first chemotherapeutic
agent to treat syphilis: 1908 • Avery, MacLeod, and McCarty
demonstrate that Griffith’s
transforming principle is DNA: 1944
• Milstein*, Kohler*, and Jeme* • Joshua Lederberg* and Edward
develop monoclonal Tatum* demonstrate the transfer
antibodies: 1975 of DNA between bacteria: 1944
• Jacob* and Monod* • Jackie Robinson baseball
• Theodor Diener demonstrates playing days: 1947–1956
• Michael Bishop* and Harold fundamental differences between demonstrate how
Varmus* discover that cancer- viroids and viruses: 1971 enzyme levels in cells
causing genes are found in are regulated: 1961
normal tissues: 1976

• Carl Woese
classifies all • Barbara McClintock*
organisms discovers transposable
into three elements in maize: 1948
domains:
1977 • Korean War: 1950–1953
• Voting age lowered • U.S. enters
from 21 to 18 in U.S.: 1971 Vietnam War: • Watson*, Crick*, Franklin,
• Equal Rights Amendment 1959 and Wilkins* determine the
• Herbert Boyer and in U.S. gives women equal
Stanley Cohen, structure of DNA: 1953
rights: 1971
clone DNA: 1973
• Arber*, Nathans*, and • D. Carlton Gajdusek* demonstrates the slow
• A retrovirus causing a rare cancer Smith* discover the infectious nature of the disease kuru, later
in humans is discovered: 1980 restriction enzyme: 1968 shown to be caused by a prion: 1957
• World Health Organization declares
smallpox is eradicated: 1980 • The first new antibiotic in 35 years
is approved by the Food and Drug
• Sony introduces the
Administration: 2000 • Largest offshore
Walkman: 1981
• Cell phone becomes • The first complete nucleotide oil rig spill in U.S.
commercially available: 1983 sequence of a bacterial history: 2010
• Attack on World
chromosome is reported: 1995
Trade Center: 2001

• First product of genetic

engineering introduced–
human insulin: 1982 • Avian influenza
• Luc Montagnier* and • The Food and Drug • Bioterrorism by anthrax
• Barry Marshall* demonstrates considered a major
Robert Gallo isolate Administration approves a spores in the U.S.: 2001
that a bacterium causes threat: 2005
ulcers: 1982 and characterize the genetically engineered
• Stanley Prusiner* isolates a human immuno- food for human
protein; a prion from a slow deficiency virus (HIV): consumption: 1994
disease infection: 1982 1983

• Kary Mullis* invents

the polymerase
chain reaction: 1983
6 Chapter 1 Humans and the Microbial World

MicroByte
Several species of naturally occurring bacteria actively degraded
at least part of the oil that gushed into the Gulf of Mexico after the
explosion of an oil rig in 2010.

Commercially Valuable Products from Bacteria

Bacteria synthesize a wide variety of different products, some of
which are commercially valuable. Although these same products
can be made in factories, bacteria often generate them faster and
cheaper. For example, different bacteria produce:
■ Cellulose: used in stereo headsets
■ Hydroxybutyric acid: used in the manufacture of dispos-
able diapers and plastics
■ Ethanol: used as a biofuel
■ Hydrogen gas: used as a possible biofuel FIGURE 1.3 Students wearing gauze masks to protect themselves
against infection with the influenza virus in 1918.
■ Oils: used as a possible biofuel
? Why might the gauze masks not protect against the influenza
■ Insect toxins: used in insecticides virus?
■ Antibiotics: used in the treatment of disease
■ Amino acids: used as dietary supplements
realized that some of these microscopic agents could cause dis-
ease, they tried to prevent their spread. Over the last 100 years,
Biotechnology
improvements in human health have been due largely to using
Biotechnology—the use of microbiological and biochemical antibiotics to treat infectious diseases, and vaccines to prevent
techniques to solve practical problems—depends on members of them. The results have been astounding!
the microbial world. The study of microorganisms led to tech- The viral disease smallpox was one of the greatest killers the
niques that allow scientists to genetically engineer plants to world has ever known, resulting in the death of approximately 10
resist a wide variety of insects, bacteria, and viruses that could million people over 4,000 years. It was brought to the New World
otherwise attack them. Biotechnology has also led to easier pro- by the Spaniards, and the devastation it caused allowed Hernando
duction of many medications such as insulin (used to treat dia- Cortez, with fewer than 600 soldiers, to conquer the Aztec
betes). In the past, insulin needed to be isolated from pancreatic Empire, whose subjects numbered in the millions. During a cru-
glands of cattle and pigs. Now it is isolated from bacteria. cial battle in Mexico City, an epidemic of smallpox raged, killing
 genetic engineering, p. 216 mainly the Aztecs who had never been exposed to the disease. In
recent times, an active worldwide vaccination program eliminated
Medical Microbiology the disease in nature, with no cases being reported since 1977.
However, the possibility that the smallpox virus could be used in
Although most microbes are beneficial or not harmful, some are
bioterrorist attacks is a great concern.
pathogens, meaning they can cause disease. In fact, more
Plague has been another major killer. One-third of the popula-
Americans died of influenza in 1918–1919 than were killed in
tion of Europe, approximately 25 million people, died of this
World War I, World War II, and the Korean, Vietnam, and Iraq
bacterial disease between 1346 and 1350. Now, less than 100
wars combined (figure 1.3). Fortunately, technological advances
people worldwide die from plague in a typical year. This dramatic
such as sanitation, vaccination, and antibiotic treatments have
decrease is primarily a result of controlling the rodent population
dramatically reduced the incidence of many of the most feared
that harbors the bacterium. In addition, the discovery of antibiotics
infectious diseases. To maintain this success, however, we must
in the twentieth century made the isolated outbreaks treatable.
educate future generations to continue their vigilance and develop
Epidemics are not limited to human populations. The great
new weapons. This is particularly important considering the rapid
famine in Ireland in the 1800s was due to a disease of potatoes. In
increase in the number of antibiotic-resistant microorganisms.
2001, a catastrophic outbreak of foot-and-mouth disease of ani-
Meanwhile, another disease, acquired immunodeficiency syn-
mals ran out of control in England. To contain this disease, one of
drome (AIDS), has risen as a modern-day plague that no vaccine
the most contagious known, almost 4 million pigs, sheep, and cat-
prevents.
tle were destroyed.
Past Triumphs
The Golden Age of Microbiology included an important period Present and Future Challenges
when scientists were learning a great deal about medically impor- Although progress has been impressive against bacterial diseases,
tant microbes. Between 1876 and 1918, most pathogenic bacteria much more still needs to be done. This is especially true for
were identified, and early work on viruses had begun. Once people the  treatment of viral infections and diseases associated with
 Part I Life and Death of Microorganisms 7

poverty. Respiratory infections and diarrheal diseases cause most without causing disease. The infected animals, however, shed
illness and deaths in the world today. Even in wealthy developed virus in urine, feces, and saliva; from there it can be inhaled by
countries with their sophisticated healthcare systems, infectious humans as an aerosol. Hantavirus disease is only one of many
diseases remain a serious threat. In the United States, about emerging human diseases associated with small animals.
750 million cases of infectious diseases occur each year, leading Some emerging diseases arise because the infectious agents
to 200,000 deaths and costing tens of billions of dollars. change abruptly and gain the ability to infect new hosts. It appears
that HIV (Human Immunodeficiency Virus), the cause of AIDS,
Emerging Diseases In addition to the long-recognized dis-
arose from a virus that once could infect only chimpanzees. The
eases, emerging diseases continue to arise (figure 1.4). These
virus causing SARS is related to viruses found in animals and may
include swine flu, severe acute respiratory syndrome (SARS),
have been transmitted from animals to humans. Some bacterial
multidrug-resistant tuberculosis, Lyme disease, hepatitis C,
pathogens differ from their normally harmless relatives in that the
acquired immunodeficiency syndrome, hemolytic uremic syn-
pathogens contain large pieces of DNA that encode toxins or other
drome, hantavirus pulmonary syndrome, mad cow disease
characteristics that allow the organism to cause disease. These
(bovine spongiform encephalopathy), and West Nile encephali-
pieces of DNA may have originated in unrelated organisms.
tis. Most of these diseases are merely newly recognized rather
Are other agents out there that may cause “new” diseases in
than actually new. Their increased occurrence and wider dis-
the future? The answer is undoubtedly yes!
tribution have brought them to the attention of health workers.
Using the latest techniques, scientists have isolated, character- Re-Emerging Diseases Infectious diseases that were under con-
ized, and identified the agents causing the diseases. Now, better trol can increase again. In some cases, the preventive measures
methods need to be developed to control them. become victims of their own success. For example, decades
A number of factors account for the emergence of diseases. of vaccination have nearly eliminated measles, mumps, and
Changing lifestyles bring new opportunities for infectious agents whooping cough in the United States, so many people have no
to spread. For instance, the suburbs of cities are expanding into firsthand knowledge of the dangers of the diseases. Couple this
rural areas, bringing people into closer contact with animals. with misinformation about vaccines, and people develop an irra-
Consequently, people become exposed to viruses and infectious tional fear of the vaccine more than the disease itself. If parents
organisms that once were far removed from their environment. A refuse to have their children vaccinated, the diseases can easily
good example is the hantavirus. This virus infects rodents, usually re-emerge.

2002
Severe acute
respiratory
syndrome (SARS)
China

1986
Bovine spongiform
1982 encephalopathy
United Kingdom 1977
E. coli O157:H7 Hantaan virus
United States 1980 Republic of Korea
1981 Hepatitis D (Delta)
1980
AIDS Italy
1989 Human T-cell
United States Hepatitis C lymphotropic virus 1
1976 United States Japan
Legionnaires’ disease
1997
United States
1992 Avian flu (H5N1)
2009 1991 Hong Kong
Venezuelan Vibrio
Swine flu cholerae 0139
Mexico hemorrhagic
fever India
1976
Cryptosporidiosis Venezuela
United States 1999
1994 Malaysian
Brazilian encephalitis
hemorrhagic Malaysia
fever 1976
Brazil Ebola 1994
hemorrhagic fever Human and equine
Zaire morbilivirus
Australia
FIGURE 1.4 “New” Infectious Diseases in Humans and Animals Since 1976 Countries where cases first appeared or were identified
appear in a darker shade.
? Why might so many of the “new” diseases first appear or be identified in the United States and Western European countries?
8 Chapter 1 Humans and the Microbial World

Diseases also re-emerge when pathogens become resistant to can be used to obtain results very quickly because they grow rap-
antimicrobial medications. Tuberculosis and malaria are re-emerging, idly and form billions of cells per milliliter on simple inexpensive
in part because they are often now more difficult to treat. growth media. In fact, most major advances made in the last cen-
Travelers and immigrants can contribute to disease re-emergence tury toward understanding life have come through the study of
by inadvertently carrying pathogens around the globe. Diseases microbes. The number of Nobel Prizes awarded to microbiologists
such as malaria, cholera, plague, and yellow fever have largely been highlights this point. Such studies constitute basic research, and
eliminated from developed countries, but they still exist in many they continue today.
parts of the world. An international traveler incubating a disease
could theoretically circle the globe, touch down in several countries, MicroAssessment 1.2
and expose many people before becoming ill. Microorganisms have an enormous impact on all living things, and
Changes in the characteristics of a population can also cause their activities are vital to human life. Microorganisms have been
re-emergence of diseases. Elderly people typically have weaker used for food production for thousands of years. They are now being
immune systems than the young, so aging populations are more used to degrade toxic pollutants and produce a variety of compounds.
susceptible to infectious agents. Individuals with AIDS are espe- Enormous progress has been made in preventing and curing infectious
cially susceptible to a wide variety of diseases, including diseases, but new ones continue to emerge and re-emerge. Microbes
tuberculosis. are model organisms, and many principles of biochemistry and
genetics have been discovered from their study.
Chronic Diseases Caused by Bacteria In addition to the dis- 4. Describe two microbial activities essential to life and three that
eases long recognized as being caused by pathogens, some ill- make our lives more comfortable.
nesses once attributed to other sources may, in fact, be caused 5. Describe three reasons why some diseases re-emerge.
by microbes. The best-known example is peptic ulcers. This 6. Why would it seem logical, even inevitable, that at least some
common affliction—once thought to be due to stress—is caused bacteria would attack the human body and cause disease? +
by a bacterium (Helicobacter pylori) and treatable with antibiot-
ics. Chronic indigestion may be caused by the same bacterium.
Infectious microbes may play important roles in other chronic
diseases as well. 1.3 ■ Living Members of the
Microbial World
Host-Microbe Interactions
All surfaces of the human body are populated with characteris- Learning Outcomes
tic communities of microorganisms, collectively called normal 5. Describe the diversity of microorganisms in terms of their numbers
microbiota, or normal flora. These organisms play a number of and ability to be grown in culture.
indispensable roles, such as preventing disease by competing with 6. Compare and contrast the Bacteria, Archaea, and Eucarya.
pathogens. In addition, members of the normal intestinal micro- 7. Compare and contrast algae, fungi, and protozoa.
biota play critical roles in the development of the intestine and 8. Explain how the scientific name of an organism is written.
the immune system, and they help degrade foods that the body
otherwise could not digest. The microbial world is incredibly diverse, with extremely small
In contrast to beneficial microbes, pathogens damage body size the only common feature. Microorganisms include bacteria,
tissues, leading to symptoms of disease. They use the human body archaea, protozoa, algae, fungi, and some multicellular parasites.
as a habitat for multiplication, persistence, and transmission to
other hosts. The disease symptoms often result from the body’s
defense mechanisms damaging the host while attempting to con- Diversity of Microorganisms
trol the pathogen. Diversity of microorganisms is evident in their appearance,
metabolism, physiology, and genetics. An interesting bit of trivia
MicroByte that highlights this diversity is the estimate that the total number
Your body carries ten times more bacterial cells (including in the
of bacterial species on this planet outnumbers mammalian species
intestinal tract) than human cells!
by a factor of 10,000! This is important to keep in mind when
describing the biodiversity (variety of different species) in any
Microorganisms as Model Organisms environment. Biodiversity is often measured by considering only
Microorganisms are wonderful model organisms to study because the number of plant and animal species, but these species actually
they display the same fundamental metabolic and genetic proper- represent only a fraction of the organisms present in any given
ties as higher life-forms. For example, all cells are composed of environment.
the same elements and they synthesize their cell structures by The diversity of microorganisms results from the fact that
similar mechanisms. They all duplicate their DNA, and when they they are the oldest forms of life and therefore have had the longest
degrade foods to harvest energy, they do so via the same metabolic time to evolve. However, they must be even more varied than
pathways. To paraphrase a Nobel Prize–winning microbiologist, what we now understand because less than 1% of all microbial
Dr. Jacques Monod—what is true of elephants is also true of bac- species can be grown and studied in the laboratory. Thus, the vast
teria, and bacteria are much easier to study. In addition, bacteria majority of microorganisms have yet to be studied.
 Part I Life and Death of Microorganisms 9

bound nucleus or any other membrane-bound organelles. Instead, the

genetic material resides in a region called the nucleoid.
Most Bacteria have specific shapes, commonly cylindrical
Lens (rod-shaped), spherical (round), or spiral. They typically have
rigid cell walls that contain peptidoglycan, a compound unique to
Specimen holder bacteria (see figure 3.31). Many of the Bacteria can move using
flagella (singular: flagellum), appendages that extend from the
cell.  bacterial shapes, p. 50  flagella, pp. 63, 74
Bacteria typically multiply by binary fission, a process in
which one cell enlarges and then divides. This forms two cells,
Focus screw
each generally identical to the original.  binary fission, p. 82

MicroByte
One group of organisms in the domain Bacteria has a membrane
surrounding its genetic material as well as other internal membranes.

Archaea
Handle Like members of the Bacteria, the Archaea have a prokaryotic cell
structure. They have similar shapes, sizes, and appearances to the
Bacteria. In addition, they multiply by binary fission, move pri-
marily by means of flagella, and have rigid cell walls. Considering
FIGURE 1.5 Model of van Leeuwenhoek’s Microscope The how much they look like Bacteria, scientists initially did not
original made in 1673 could magnify the object being viewed almost believe they could be so different chemically. We now know,
300 times. The object being viewed is brought into focus with the
however, that there are major differences between the Bacteria
adjusting screws. Note the small size.
and the Archaea, and that Archaea are more closely related to
humans than they are to Bacteria. In fact, you are more closely
related to a plant than Archaea are to Bacteria!
The living microbial world includes the kinds of cells that van Archaea and Bacteria have important differences in their
Leeuwenhoek observed looking through his simple microscope chemical composition. For example, the archaeal cell wall does
(figure 1.5). Although he could not realize it at the time, members not contain peptidoglycan, whereas the bacterial wall does. One of
of the microbial world, in fact, all living organisms, can be classi- the most significant distinctions is in their ribosomal RNA
fied into three distinct domains—Bacteria, Archaea, and Eucarya. sequence, a characteristic discussed in other chapters. ribosomal
RNA, p. 246
Organisms in each domain share properties that distinguish
them from members of the other domains. Many characteristics, An interesting feature of many of the Archaea is their ability
however, are common among members of different domains. to grow in extreme environments that kill most other organisms.
Table 1.1 compares members of the domains Bacteria, Archaea, Some, for example, grow in salt concentrations 10 times higher
and Eucarya. than that of seawater. These organisms grow in such habitats as
the Great Salt Lake and the Dead Sea. Others grow best at
extremely high temperatures. One member can grow at a tem-
perature of 121°C! (100°C is the temperature at which water
Bacteria boils at sea level). Some Archaea can be found in the hot springs
Members of the domain Bacteria are prokaryotes, meaning they are at Yellowstone National Park.
single-celled organisms consisting of a prokaryotic cell (meaning While the Archaea that grow in extreme environments are the
“prenucleus”). This cell type typically does not contain a membrane- most intensively studied, others are abundant in moderate

TABLE 1.1 Comparison of Traits of Typical Bacteria, Archaea, and Eucarya

Bacteria Archaea Eucarya

Size 0.3–2 μm 0.3–2 μm 5–50 μm

Nuclear membrane No No Yes
Cell wall Peptidoglycan present No peptidoglycan No peptidoglycan
Membrane-bound organelles No No Yes
Where found In all environments In all environments In environments that are not extreme
10 Chapter 1 Humans and the Microbial World

environments. They are widely distributed in soils, the oceans,

marshes, as well as in the intestinal tract. Some have been impli-
cated in severe cases of periodontitis, a destructive inflammation
of the gums.

Eucarya
Members of the Eucarya are eukaryotes, meaning they are com-
posed of one or more eukaryotic cells (“true nucleus”). These
cells have a membrane-bound nucleus and other organelles, mak-
ing them far more complex than prokaryotes.
The microbial members of the Eucarya include algae (sin-
gular: alga), fungi (singular: fungus), and protozoa (singular:
protozoan). Algae and protozoa are also referred to as protists.
Some multicellular parasites are considered in this text because
FIGURE 1.6 Alga Micrasterias, a green alga composed of two sym-
they kill millions of people around the world, especially in metrical halves (100×).
developing nations. This group of organisms, helminths, include
roundworms and tapeworms. The eukaryotic members of the ? What general features of algae distinguish them from other
eukaryotic microorganisms?
microbial world are compared in table 1.2.

Algae Protozoa
Algae are a diverse group of photosynthetic eukaryotes. Some are Protozoa are a diverse group of microscopic, single-celled organ-
single-celled, whereas others are multicellular, and many different isms that live in both aquatic and terrestrial environments.
size and shapes are represented (figure 1.6). They all contain chlo- Although microscopic, they are very complex organisms and gen-
roplasts, many of which have chlorophyll, a green pigment. Some erally much larger than prokaryotes (figure 1.8). Unlike algae and
also contain other pigments that give
them characteristic colors. The pig-
ments absorb the energy of light, which
is used in photosynthesis. Algae are
usually found near the surface of either
salt or fresh water. Their cell walls are
rigid, but their chemical composition is
quite distinct from that of the Bacteria
and Archaea. Many algae move by
means of flagella, which are structur- Reproductive
structures
ally far more complex and unrelated to (spores)
flagella of prokaryotes.

Fungi
Fungi are another diverse group of
eukaryotes. Some are single-celled Mycelium
yeasts, but many are large multicellular
organisms such as molds and mush-
rooms (figure 1.7). In contrast to algae,
fungi gain their energy from degrading (a) 10 µm (b) 10 µm
organic materials and are found wher-
ever organic materials are present. FIGURE 1.7 Fungi (a) Yeast, Malassezia furfur. (b) Aspergillus, a typical mold form whose repro-
ductive structures rise above the mycelium.
Unlike algae, which live primarily in
water, fungi live mostly on land. ? What type of cells make up molds and mushrooms?

TABLE 1.2 Comparison of Eukaryotic Members of the Microbial World

Algae Fungi Protozoa

Cell organization Single- or multicellular Single- or multicellular Single-celled

Source of energy Sunlight Organic compounds Organic compounds
Size Microscopic or macroscopic Microscopic or macroscopic Microscopic
 Part I Life and Death of Microorganisms 11

MicroAssessment 1.3
Three domains of life exist: Bacteria, Archaea, and Eucarya.
Members of the Bacteria and Archaea are prokaryotes, but they
are distinctly different from each other in certain features of their
chemical composition. Members of the Eucarya are eukaryotes;
algae, fungi, protozoa, and multicellular parasites belong to this
group. Organisms are named according to the Binomial System of
Nomenclature.
7. Name one feature that distinguishes members of the Bacteria
from the Archaea.
8. List two features that distinguish prokaryotes from eukaryotes.
9. The binomial system of classification uses both a genus and a
species name. Why are two names used? +

20 µm 1.4 ■ Non-Living Members

FIGURE 1.8 Protozoan A paramecium moves with the aid of hair-
like appendages on the cell surface.
of the Microbial World
? How do protozoa differ from both fungi and algae? Learning Outcome
fungi, protozoa do not have a rigid cell wall. Most protozoa ingest 9. Compare and contrast viruses, viroids, and prions.
organic compounds as food sources and are motile.
Viruses, viroids, and prions are not living; they are acellular infec-
Nomenclature tious agents. By definition, an organism must be composed of one
In biology, the Binomial System of Nomenclature refers to a two- or more cells to be alive. Non-living agents are not microorgan-
word naming system. The first word in the name indicates the isms, so the term microbe is often used as a general term to
genus, with the first letter always capitalized; the second indicates include any member of the microbial world.   viroids, p. 328
the specific epithet, or species name, and is not capitalized. Both  prions, p. 328
words are usually italicized or underlined—for example, Viruses consist of nucleic acid packaged within a protein
Escherichia coli. The genus name is commonly abbreviated, with coat, and come in a variety of shapes (figure 1.9). To multiply,
the first letter capitalized—as in E. coli. A number of different spe- viruses use the machinery and nutrients of living cells, referred to
cies are included in the same genus. Members of the same species as hosts. Outside the hosts, however, viruses are inactive. Thus,
may vary from one another in minor ways, but not enough to give viruses are obligate intracellular parasites. All forms of life
the organisms different species names. These differences, how- including members of the Bacteria, Archaea, and Eucarya can be
ever, may result in the organism being given different strain desig- infected by viruses but of different types. Although viruses fre-
nations, for example, E. coli strain B or E. coli strain K12. quently kill the cells in which they multiply, some types can
Bacteria are often referred to informally by names resembling remain within the host cell without causing obvious ill effects. As
genus names but are not italicized. For example, species of the host cells multiply, they copy the viral genetic information,
Staphylococcus are often called staphylococci. passing it along to their progeny.

Nucleic acid
Protein coat

(a) 50 nm (b) Protein coat Nucleic acid Tail 50 nm (c) Nucleic acid 50 nm

FIGURE 1.9 Viruses That Infect Three Kinds of Organisms (a) Tobacco mosaic virus that infects tobacco plants. A long hollow protein
coat surrounds a molecule of nucleic acid. (b) A bacterial virus (bacteriophage), which infects bacteria. Nucleic acid is surrounded by a protein coat
(head) which is attached to a hollow protein tail. (c) Influenza virus. This virus infects humans and causes flu.
? Why can viruses be so much smaller than cells and still replicate?
12 Chapter 1 Humans and the Microbial World

Distinguishing Characteristics
TABLE 1.3
of Viruses, Viroids, and Prions
Viruses Viroids Prions

PSTV Obligate Obligate Abnormal form of a

intracellular intracellular cellular protein
agents agents
Consist of either Consist only of Consist only of
DNA or RNA, RNA; no protein protein; no DNA or
surrounded by a coat RNA
protein coat

T7 DNA Prions are infectious proteins responsible for several fatal

neurodegenerative diseases in humans and other animals
(figure 1.11). They apparently are simply misfolded versions of
normal cellular proteins found in the brain. When the misfolded
PSTV version contacts the normal cellular protein, it forces the normal
protein to misfold, thereby resulting in cells becoming filled with
1 µm
abnormal proteins that bind together to form threadlike structures
called fibrils. The fibril-filled cells are not able to function. The
FIGURE 1.10 The Size of a Viroid Compared with a Molecule disease is acquired by eating prion-containing nervous tissues.
of DNA from a Virus That Infects Bacteria (T7) The red arrows
point to potato spindle tuber viroids (PSTV); the blue arrow points to
Prions are more resistant to degradation by cellular enzymes than
the bacterial virus T7 DNA. are their normal counterparts. Prions are also resistant to the usual
sterilization procedures that destroy viruses and bacteria. Some
? How does a viroid differ from a virus?
scientists speculate that Alzheimer’s and Parkinson’s diseases are
Viroids are simpler than viruses, consisting of only a single, caused by mechanisms similar to those of prion diseases.
short piece of ribonucleic acid (RNA) (figure 1.10). Like viruses, The distinguishing features of the non-living members of the
they multiply only inside cells. Viroids cause a number of plant dis- microbial world are listed in table 1.3. The relationships of the
eases, and some scientists speculate that they may cause diseases in major groups of the microbial world to one another are presented
humans, although no evidence for this yet exists. in figure 1.12.

MicroAssessment 1.4
Viruses, viroids, and prions are infectious agents.
10. Describe the chemical composition of viruses, viroids, and
prions.
11. Which of the non-living members of the microbial world seems to
be the least threat to human health? +

1.5 ■ Size in the Microbial World

Learning Outcome
10. Compare the size differences among microbes.

Although all members of the microbial world are small, microbes

span a tremendous range of sizes—the largest eukaryotic cells are
about a million times bigger than the smallest viruses (figure 1.13).
Even within a single group, wide variations exist. For example,
Bacillus megaterium and Mycoplasma pneumoniae are both bac-
teria, but the former are much larger. The variation in size of
bacteria was recently expanded when a bacterium longer than
0.5 mm was discovered (see Perspective 1.1). It is so big, in fact,
50 nm
that it is visible to the naked eye. More recently, an even larger
FIGURE 1.11 Prion Prions isolated from the brain of a scrapie- bacterium, round in shape, was discovered. Its volume is 70 times
infected hamster. This neurodegenerative disease is caused by a prion. larger than the previous record holder. Likewise, a eukaryotic cell
? Why are prions visible here when normal cellular proteins are not? was recently discovered that is not much larger than a typical
Focus Figure
Microbial World

Organisms Infectious agents

(living) (non-living)

Domain Bacteria Archaea Eucarya Viruses Viroids Prions

Prokaryotes (unicellular) Eukaryotes

Algae Protozoa Fungi Helminths

(unicellular or (unicellular) (unicellular or (multicellular
multicellular) multicellular) parasites)

Protists

FIGURE 1.12 The Microbial World Although adult helminths are generally not microscopic, some stages in the life cycle of many disease-
causing helminths are.
? The members of which two domains cannot be distinguished microscopically?

Nucleus

Small Proteins Viruses Mitochondria

molecules
Prion fibril

Atoms Lipids Ribosomes Smallest Most Most eukaryotic cells Adult roundworm
bacteria bacteria

Human height
Electron microscope

Light microscope

Unaided human eye

0.1 nm 1 nm 10 nm 100 nm 1 µm 10 µm 100 µm 1 mm 1 cm 0.1 m 1m 10 m

The basic unit of length is the meter (m), and all These units of measurement correspond to units
other units are fractions of a meter. in an older but still widely used convention.

nanometer (nm) = 10–9 meter = .000000001 meter 1 angstrom (Å) = 10–10 meter
micrometer (µm) = 10–6 meter = .000001 meter 1 micron (µ) = 10–6 meter
millimeter (mm) = 10–3 meter = .001 meter
1 meter = 39.4 inches

FIGURE 1.13 Sizes of Molecules, Non-Living Agents, and Organisms

? Why is a logarithmic scale necessary when comparing sizes of members of the microbial world?
13
14 Chapter 1 Humans and the Microbial World

PERSPECTIVE 1.1
Every Rule Has an Exception
We might assume that because prokaryotes which means “sulfur pearl of Namibia”
have been so intensively studied over the past (figure 2). Although scientists were initially
hundred years, no major surprises are left skeptical that prokaryotes could be so large,
to be discovered. This, however, is far from there is no question in their minds now.  
the truth. In the mid-1990s, a large, peculiar- Thiomargarita, p. 273
looking organism was seen when the intestinal In contrast to the examples of large bac-
tracts of certain fish from both the Red Sea in teria, a cell found in the Mediterranean Sea
the Middle East and the Great Barrier Reef in is 1 μm in width. It is a eukaryote because
Australia were examined. This organism, named it contains a nucleus even though it is about
Epulopiscium cannot be cultured in the labora- the size of a typical bacterium.
tory (figure 1). Its large size, 600 μm long and How small can an organism be? An answer
80 μm wide, made it clearly visible without to this question may be at hand as a result of a
any magnification, and suggested that this new microbe discovered off the coast of Iceland.
organism was a eukaryote. However, it did not The organism, found in an ocean vent where the 1 µm
have a membrane-bound nucleus. A chemical temperature was close to the boiling point of FIGURE 3 Five Cells of Nanoarchaeum
analysis of the cell confirmed that it was a pro- water, cannot be grown in the laboratory by equitans, Attached on the Surface of
karyote and a member of the domain Bacteria. itself, but grows attached to another much larger the (Central) Ignicoccus Cell Platinum
This very long, slender organism is an exception member of the Archaea (figure 3). These tiny shadowed.
to the rule that prokaryotes are always smaller organisms, also members of the Archaea, have
than eukaryotes.  Epulopiscium, p. 272 been named Nanoarchaeum equitans, which
In 1999, a prokaryote even larger in volume means “riding the fire sphere.” The organism to
was isolated from the muck of the ocean floor which N. equitans is attached is Ignicoccus, contained in N.  equitans is less than in any
off the coast of Namibia in Africa. It is a spher- which means “fire ball.” Ignicoccus grows very known organism, and only about one-tenth the
ical organism 70 times larger in volume than well without its rider. N. equitans is spherical amount found in the common gut organism,
Epulopiscium. Since it grows on sulfur com- and only about 400 nanometers in diameter, Escherichia coli. Further analysis of these cells
pounds and contains glistening globules of sul- about a quarter the diameter of Ignicoccus. suggests that they may resemble the earliest
fur, it was named Thiomargarita namibiensis, Also, the amount of genetic information (DNA) cells and therefore the ancestor of all life. The
scientists who discovered N. equitans suggest
that many more unusual organisms related to
N. equitans will be discovered. They are prob-
ably right!  Nanoarchaeum, p. 279
Not only have surprises been found in the
sizes of prokaryotes, but equally unexpected
observations have been made regarding their
Epulopiscium internal structures. A major distinction between
(prokaryote) prokaryotes and eukaryotes is the lack of a
nuclear membrane in prokaryotes (see table 1.1).
However, a recent microscopic study on one
Paramecium
(eukaryote)
group in the domain Bacteria, the Planctomyces,
has clearly shown that the nucleoid is sur-
rounded by a membrane. Further, these bacteria
carry out endocytosis, a mechanism for the
0.1 mm 0.2 mm uptake of foreign material that previously was
FIGURE 1 Longest Known Bacterium, FIGURE 2 Thiomargarita namibiensis associated only with eukaryotes. All of these
Epulopiscium Note how large this prokary- The average Thiomargarita namibiensis is two- exceptions to long-standing rules point out the
ote is compared with the four eukaryotic tenths of a millimeter, but some reach three need to keep an open mind and not jump to
paramecia in the photo. times that size. conclusions.

bacterium. These, however, are rare exceptions to the rule that MicroAssessment 1.5
eukaryotes are larger than prokaryotes, which in turn are larger The size range of microbes is tremendous. As a general rule,
than viruses. eukaryotes are the largest and obligate intracellular parasites the
The small size of microbes requires measurements not com- smallest.
monly encountered in everyday life. Logarithms are enormously 12. Place in order with respect to typical size (arrange from smallest
helpful, especially in designating the sizes of prokaryotes and to largest) bacteria, eukaryotic cells, and viruses.
viruses. A brief discussion of measurements and logarithms is 13. What factor limits the size of free-living cells? +
given in Appendix I.
 Part I Life and Death of Microorganisms 15

FUTURE CHALLENGES 1.1

Entering a New Golden Age
For all the information that has been gathered the finding of 1,800 previously unknown bac- living organisms exist whose chemical structure
about the microbial world, it is remarkable how terial species. This estimate is based on the is not based on the carbon atom? Will living
little we know about its prokaryotic members. analysis of the DNA isolated from these microor- organisms be found whose genetic information
This is not surprising in view of the fact that ganisms. The biodiversity of the microbial world is coded in a chemical other than deoxyribo-
less than 1% of the prokaryotes have ever been is astounding! nucleic acid? What new metabolic pathways
studied. In large part, this is because only one Exploring the unknowns in the micro- remain to be discovered? As extreme environ-
in a hundred of the prokaryotes in the environ- bial world is a major challenge and should ments are mined for their living biological
ment can be cultured in the laboratory. Part of answer many intriguing questions fundamental diversity, there seems little doubt that many
the current revolution in microbiology, however, to understanding the biological world. What are surprises will be found. In many cases these sur-
will allow us to inventory the millions of species the extremes of temperature, salt, pH, radioac- prises will be translated into new biotechnology
yet to be discovered. This is now being done. tivity, and pressure in which prokaryotes can products on this planet, and they will help shape
Using techniques that helped decipher the human live? Are there organisms growing in even the way we look for life on other planets.
genome, scientists have begun to analyze the bio- more extreme environments? If life can exist on One hundred years ago we were in the
logical content of the oceans. In a small volume this planet under such extreme conditions, what Golden Age of Microbiology. With new
of water from the Sargasso Sea—an area of the does this mean about the possibility of finding technologies available, we are entering a
ocean that contains few nutrients and therefore living organisms on other planets? Although second Golden Age of identifying unknown
presumably few organisms—scientists reported considered highly unlikely, is it possible that members of the microbial world.

Summary
1.1 ■ The Dispute Over Spontaneous Generation new lifestyles and changing infectious agent (figure 1.4). Diseases once
The belief in spontaneous generation was challenged by Francesco under control re-emerge when successful preventive measures become
Redi in the seventeenth century. victims of their own success, pathogens become resistant to antimicro-
bial medications, travelers and immigrants carry pathogens around the
Early Experiments globe, and populations age or otherwise become more susceptible to
The experiments of Needham supported the idea of spontaneous genera- diseases. People have become lax in having children vaccinated. The
tion while those of Spallazani did not. normal microbiota plays a protective role, but pathogens cause disease.
Experiments of Pasteur Microorganisms as Model Organisms
The experiments of Louis Pasteur disproved spontaneous generation Microorganisms are excellent model organisms because they grow
(figure 1.1). rapidly on simple, inexpensive media. Their growth reveals the same
genetic, metabolic, and biochemical principles as higher organisms.
Experiments of Tyndall
1.3 ■ Living Members of the Microbial World (table 1.1)
John Tyndall showed that some microbial forms are not killed by boil-
ing. He and Ferdinand Cohn discovered endospores, the heat-resistant Diversity of Microorganisms
forms of bacteria. The microbial world is incredibly diverse (figure 1.12).
Bacteria
1.2 ■ Microbiology: A Human Perspective
Members of the Bacteria are single-celled prokaryotes that have pep-
Vital Activities of Microorganisms tidoglycan in their cell wall.
The activities of microorganisms are vital for the survival of all
Archaea
other organisms, including humans. Bacteria convert the nitro-
Members of the Archaea are identical in appearance to the Bacteria, but
gen gas in air into a form that plants and other organisms can
are very different in their chemical composition. They do not contain
use. Microorganisms replenish O2 and degrade certain materials
peptidoglycan. Many Archaea grow in extreme environments.
that no other organisms can.
Eucarya
Applications of Microbiology Members of the Eucarya have eukaryotic cell structures (table 1.2).
Bread, wine, beer, and cheeses are made today using technology devel- Algae can be single-celled or multicellular, and use sunlight as an
oped 4,000 years ago. Bacteria are used to degrade toxic pollutants and energy source (figure 1.6). Fungi include single-celled yeasts and mul-
to synthesize a variety of different products, such as cellulose, hydroxy- ticellular molds and mushrooms; they use organic compounds as food
butyric acid, ethanol, antibiotics, and amino acids. Biotechnology (figure 1.7). Protozoa are typically motile single-celled organisms that
depends on members of the microbial world. use organic compounds as food (figure 1.8).
Medical Microbiology Nomenclature
Many devastating diseases such as smallpox, plague, and influenza Organisms are named according to a binomial system. Each organism
have determined the course of history. Diseases emerge because of has a genus and a species name, written in italics or underlined.
16 Chapter 1 Humans and the Microbial World

1.4 ■ Non-Living Members of the Microbial World 1.5 ■ Size in the Microbial World
The non-living members of the microbial world are not composed Although they are all small, sizes of members of the microbial world vary
of cells (table 1.3). Viruses consist of nucleic acid within a protein tremendously (figure 1.13).
coat (figure 1.9). Viroids consist of a single, short RNA molecule
(figure 1.10). Prions consist only of protein; apparently, they are mis-
folded versions of normal cellular protein (figure 1.11).

Review Questions
Short Answer 6. The Archaea
1. How did Louis Pasteur help disprove spontaneous generation? 1. are microscopic.
2. Give three reasons why life could not exist without the activities of 2. are commonly found in extreme environments.
microorganisms. 3. contain peptidoglycan.
3. List five beneficial applications of bacteria. 4. contain mitochondria.
4. State three reasons why there is a resurgence of infectious diseases 5. are most commonly found in the soil.
today. a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5
5. Name the prokaryotic groups in the microbial world. 7. Prokaryotes typically do not have
6. Name one location where you could isolate members of the a) cell walls. b) flagella. c) a nuclear membrane.
Archaea. d) specific shapes. e) genetic information.
7. How might you distinguish a prokaryotic cell from a eukaryotic cell?
8. Nucleoids are associated with
8. In the designation Escherichia coli B, what is the genus? What is
1. genetic information. 2. prokaryotes.
the species? What is the strain?
3. eukaryotes. 4. viruses. 5. prions.
9. Why are viruses not microorganisms?
a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5
10. Name three non-living groups in the microbial world and describe
their major properties. 9. Viruses
1. contain both protein and nucleic acid.
Multiple Choice 2. infect all domains of life.
1. The property of endospores that led to confusion in the experi- 3. can grow in the absence of living cells.
ments on spontaneous generation is their 4. are generally the same size as prokaryotes.
a) small size. 5. always kill the cells they infect.
b) ability to pass through cork stoppers. a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5
c) heat resistance. 10. Antony van Leeuwenhoek could not have observed
d) presence in all infusions. a) roundworms. b) Escherichia coli.
e) presence on cotton plugs. c) yeasts. d) viruses.
2. The “Golden Age of Microbiology” was the time when
a) microorganisms were first used to make bread. Applications
b) microorganisms were first used to make cheese. 1. The American Society for Microbiology is preparing a “Microbe-
c) most pathogenic bacteria were identified. Free” banquet to emphasize the importance of microorganisms in
d) a vaccine against influenza was developed. the diet. What foods could not be on the menu?
e) antibiotics became available. 2. If you were asked to nominate one of the individuals mentioned in
3. Microorganisms play a role in this chapter for the Nobel Prize, who would it be? Make a state-
ment supporting your choice.
a) disease. b) biodegradation. c) cheese production.
d) nitrogen recycling. e) all of the above.
Critical Thinking +
4. Which disease was once thought to be due to stress but is now
known to be caused by a bacterium? 1. A microbiologist obtained two pure biological samples: one of
a virus, and the other of a viroid. Unfortunately, the labels had
a) smallpox b) peptic ulcers c) AIDS
been lost. The microbiologist felt she could distinguish the two
d) plague e) influenza by analyzing for the presence or absence of a single molecule.
5. The prokaryotic members of the microbial world include What molecule would she search for and why?
1. algae. 2. fungi. 3. prions. 4. bacteria. 5. archaea. 2. Why is the bacterium that causes anthrax such an effective agent
a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5 of bioterrorism?
2 The Molecules of Life
KEY TERMS
Amino Acid A subunit of a protein.
Atom The basic unit of matter.
Carbohydrate A compound
Lipid An organic molecule that is
insoluble in water.
Macromolecule A very large
molecule usually consisting of
containing principally carbon, repeating subunits.
hydrogen, and oxygen in a ratio
of 1:2:1. Molecule Two or more atoms held
together by chemical bonds.
Covalent Bond A strong chemical
bond formed when two atoms share Nucleic Acid A macromolecule
electrons. consisting of chains of nucleotide
subunits to form either DNA or
Hydrogen Bond The attraction
RNA.
between a hydrogen atom in a polar
molecule and an electronegative Nucleotide The subunit of nucleic
atom in the same or another polar acids.
molecule. Organic Compound A compound
Inorganic Compound A that has a carbon atom covalently
compound that is not organic. bonded to a hydrogen atom.
Ion An atom or molecule that has pH A measure of the hydrogen ion
gained or lost one or more electrons. concentration or acidity of a solution
on a scale of 0 to 14.
Ionic Bond A chemical bond
resulting from the attraction between Protein A macromolecule
Ball-and-stick model of water molecules. consisting of one or more chains
positively and negatively charged
ions; ions form when electrons of amino acids.
leave one atom and become part
A Glimpse of History of the other.
Louis Pasteur (1822–1895) is often considered the father of bacteriology
because of his many contributions to the field. However, he started his
scientific career as a chemist, researching crystals for the French wine
industry. In his studies, he worked with tartaric and paratartaric acids,

T
 
which form thick crusts within wine barrels. These two substances form o understand how cells interact with one another and
crystals that are identical in their chemical composition, but affect polar- their environment, you must be familiar with the com-
ized light very differently. When polarized light passes through tartaric ponents that make up all living matter. The principles
acid crystals, the light rotates (twists) to the right. In contrast, paratar- described in this chapter are fundamental to information through-
taric acid crystals have no effect on the light. Pasteur wanted to learn
out the text.
how the crystals differed.
Pasteur noticed that under a microscope, all tartaric acid crystals
looked identical, whereas paratartaric acid crystals had two kinds of struc-
tures. Using tweezers, Pasteur carefully separated the two types of paratar-
taric acid crystals and dissolved them in separate flasks of water. He found 2.1 ■ Atoms and Elements
that one solution rotated polarized light to the left and the other, to the
right. Solutions containing equal numbers of the two crystals did not rotate Learning Outcome
the light. Pasteur concluded that paratartaric acid must be a mixture of two
1. Describe the characteristics of atoms and elements and how atoms
types of chemical structures, each being the mirror image, or stereoisomer,
can be depicted diagrammatically.
of the other. The stereoisomers counteracted the light-rotating effects of
each other and, as a result, the mixture did not rotate the light.
What Pasteur studied as a simple problem in chemistry has impli- Atoms, the basic units of all matter, are made up of three major
cations far beyond what he ever imagined. In fact, stereoisomers of the components (figure 2.1):
same molecule can have very different properties. For example, one
form of a key ingredient in the artificial sweetener aspartame is sweet ■ Neutrons: uncharged particles
but the other form is bitter. Pasteur’s work demonstrates that seemingly ■ Protons: positively charged particles
unimportant observations sometimes lead to significant discoveries with
far-reaching implications. ■ Electrons: negatively charged particles

17
18 Chapter 2 The Molecules of Life

6e–
6p+
Proton 6n0

Neutron (a)

Mass
number 12

Cloud of
Atomic 6 C Element
symbol
number
electrons

(b)

FIGURE 2.1 Atom A proton has a positive charge, a neutron has a

neutral charge, and an electron has a negative charge. The electrons
move around the nucleus as a cloud, arranged in shells of different
energy levels. C
? How does the number of electrons in an atom compare to the
number of protons?
(c)
The number of protons normally equals the number of electrons,
FIGURE 2.2 Depictions of an Atom An atom of carbon (C) is
so the atom as a whole has no charge. The neutrons and protons shown in three different ways: (a) cloud structure, (b) symbol designa-
make up the heaviest part of the atom—the nucleus—and the tion, and (c) Lewis structure.
electrons form an electron “cloud” around the nucleus (figure 2.1).
? How would the Lewis structure of hydrogen be different from
An atom is distinguished by its atomic number, the number that of carbon?
of protons in its nucleus (table 2.1). For example, a hydrogen
atom has one proton so its atomic number is 1; a carbon atom has naturally occurring elements, but living material is primarily made
6 protons so its atomic number is 6. Each atom has a mass num- up of only four: carbon (the symbol is C), hydrogen (H), oxygen
ber, the sum of the number of protons and neutrons (electrons are (O), and nitrogen (N). Two other elements, phosphorus (P), and
too light to contribute to the mass). A hydrogen atom that has 1 sulfur (S), are also found in all living systems. All atoms of an
proton and no neutrons has both an atomic number and mass element have the same atomic number, but they can have different
number of 1. A carbon atom that has 6 protons and 6 neutrons mass numbers. That is, they have the same number of protons but
has an atomic number of 6 and a mass number of 12 (figure 2.2). can have different numbers of neutrons. Nearly 99% of naturally
The symbols for atoms are sometimes written with the atomic occurring carbon atoms have 6 neutrons, but a few have 7 or 8.
number in subscript to the left, and the mass number in super- These various forms—isotopes—are useful tools in biological
script to the left. research (see Perspective 2.1).
An element consists of only one kind of atom and cannot be Electrons of an atom are arranged in shells around the nucleus,
separated into simpler parts by chemical methods. There are 92 with each shell having a limit to the number of electrons it can hold.

TABLE 2.1 Characteristics of Atoms Common in Biology

Number of
Atomic Number Mass Number Covalent Bonds Approximate %
Atom Symbol (Number of Protons) (Protons + Neutrons) Formed in Cells

Hydrogen H 1 1 1 49
Carbon C 6 12 4 25
Nitrogen N 7 14 3 0.5
Oxygen O 8 16 2 25
Phosphorus P 15 31 3 0.1
Sulfur S 16 32 2 0.4
 Part I Life and Death of Microorganisms 19

PERSPECTIVE 2.1
Isotopes: Valuable Tools for the Study of Biological Systems
One important tool in the analysis of living only difference is that the molecules containing other molecules. Tritiated uridine will label
cells is the use of isotopes, variant forms tritium can be detected by the radiation they RNA (uridine is a component of RNA).
of atoms that have different atomic masses. emit. Radioisotopes are also used in medical diag-
The nuclei of certain atoms can have greater Radioisotopes are used in numerous ways nosis. For example, to evaluate proper func-
or fewer neutrons than usual and thereby be in biological research. They are frequently tioning of the human thyroid gland—which
heavier or lighter than typical. For example, added to growing cells in order to label par- produces the iodine-containing hormone
the most common form of the hydrogen atom ticular molecules, thereby making them thyroxin—doctors often administer radioac-
contains one proton and no neutrons and has detectable. For example, tritiated thymidine tive iodine. He or she would then scan the
a mass number of 1 (1H). Another form also (thymidine is a component of DNA) added to gland later to determine if the amount and
exists in nature in very low amounts. This iso- growing bacteria will label only DNA and no distribution of iodine is normal (figure 1).
tope, 2H (deuterium), contains one neutron. A
third, even heavier isotope, 3H (tritium), is not
found in nature but can be made by a nuclear
reaction in which stable atoms are bombarded
with high-energy particles. This latter isotope
is unstable and gives off radiation (decays) in
the form of rays or electrons, which can be very
sensitively measured by a radioactivity coun-
ter. Once the atom has finished disintegrating,
it no longer gives off radiation and is stable.
The other properties of radioactive isotopes,
called radioisotopes, are very similar to their
non-radioactive counterparts. For example, tri-
(a) (b)
tium combines with oxygen to form water and
with carbon to form hydrocarbons, and both Figure 1 Radioisotopes (a) Physicians use scintillation counters such as this to detect
molecules have biological properties similar to radioisotopes. (b) A scan of the thyroid gland 24 hours after the patient received radioactive
those of their non-radioactive counterparts. The iodine.

The electrons are attracted to the protons in the nucleus, so shells MicroAssessment 2.1
closer to the nucleus are filled before electrons occupy other shells.
The basic unit of all matter, the atom, is composed of protons,
The first shell (closest to the nucleus) holds no more than 2 elec-
electrons, and neutrons. The four most important elements in biology
trons. These electrons are most highly attracted to the nucleus and are carbon, hydrogen, oxygen, and nitrogen. In diagrams, atoms can
have the lowest energy levels in that particular atom. Once that shell be shown as a cloud structure, symbol designation, or Lewis structure.
is filled, additional electrons occupy the next shell, to a maximum
1. Why are electrons not considered in determining the mass
of 8 electrons. Larger atoms have additional shells that hold even number of an element?
more electrons. Most atoms of biological significance follow the
2. What is the “octet rule” and its biologically important exception?
“octet rule,” meaning they are most stable when their outer shell has
3. Why is the energy level of an electron higher the farther it is
8 electrons. An important exception is hydrogen, with a single shell;
from the nucleus? +
recall that the first shell has a limit of 2 electrons.
Valence electrons are those found in an atom’s outer shell.
They are particularly important in chemical bond formation, which
will be discussed next. Lewis structures are often used to show 2.2 ■ Chemical Reactions
atoms in a way that highlights the number of valence electrons and Bonds
(figure 2.2c). Each of the valence electrons is illustrated as a dot
surrounding the atomic symbol; any other electrons are ignored. Learning Outcomes
2. Describe the importance of valence electrons.
3. Compare and contrast ionic bonds, covalent bonds, and hydrogen
MicroByte bonds.
If the nucleus of an atom were a marble in the center of a football 4. Explain the concept of a mole and why molarity is important in
field, the electrons would occupy the entire space of the stadium. chemistry.
20 Chapter 2 The Molecules of Life

e– Covalent Bonds
Na Cl Na Cl Atoms often share pairs of valence electrons, the basis for
covalent bond formation. One pair of shared electrons is a cova-
lent bond. The number of covalent bonds an atom can form—its
valence—is the number of electrons the atom must gain or lose to
Na+ Cl – Na+ Cl – fill its outer shell (table 2.1). Two or more atoms joined together
by covalent bonds form a molecule. The atoms that make up a
molecule may be of the same or different elements. H2 is a mol-
ecule of hydrogen gas formed from two atoms of hydrogen; a
Na+ Cl – water molecule (H2O) contains two hydrogen atoms bonded to one
oxygen atom. Water is a compound, a molecule consisting of
(a) more than one element.
Carbon (C)—a critical atom in biology—frequently forms
covalent bonds. This element has four electrons in its outer shell,
and needs four more to fill it. A hydrogen (H) atom has one elec-
tron and requires an additional one to fill its shell. If one C atom
shares electrons with four H atoms, methane (CH4) is created
(figure 2.4; see also table 2.1). Carbon atoms joined by covalent
(b)
bonds to H atoms form an organic compound. All other com-
FIGURE 2.3 Ions and Ionic Bonds (a) Lewis model of sodium pounds are inorganic compounds.
and chloride ions being formed, and an ionic bond between them.
(b) Space-filling model of a salt crystal being formed by ionic bond-
A covalent bond is indicated by a dash between the two atoms
ing. Note that by convention, each type of atom is depicted using a sharing the pair of electrons; for example C–H. Sometimes two pairs
consistent color. Also, an atom gets smaller when it loses an electron of electrons are shared. This forms a double covalent bond indicated
and larger when it gains one. by two lines between the atoms; for example O=C=O (CO2).
? Which of the ions in this figure is an anion, and which is a cation? Covalent bonds are strong. Double bonds are stronger than single
bonds, and triple bonds are stronger still. The stronger the bond, the
more difficult it is to break. Consequently, covalent bonds do not
Atoms lose, gain, or share the electrons in their outer shell to break unless exposed to certain chemicals or large amounts of energy,
achieve the most stable state. This is the basis for chemical bond generally as heat. The temperatures required are incompatible with
formation. Recall that with the exception of hydrogen, biologi-
cally important atoms are most stable when they have eight
valence electrons (the octet rule).
H
4 H + C H C H
Ions and Ionic Bonds
Each hydrogen atom Carbon needs H
An atom that gains or loses an electron is no longer neutral—it is an needs one electron four electrons Methane
ion (figure 2.3). Positively charged ions are called cations; nega- to fill its valence. to fill its valence.
tively charged ions are anions. Atoms that gain an electron become
(a)
negatively charged, whereas those that lose an electron become posi-
tively charged. The type and amount of charge are indicated by a
superscript to the right of the chemical symbol. For example, Na+
indicates a sodium atom that has lost one electron and therefore car- H
ries a +1 (positive) charge; Mg2+ indicates a magnesium atom that has
lost two electrons and therefore has a +2 charge.
H C H CH4
Ionic bonds form between cations and anions because of H
strong attractions between positive and negative charges
Each line represents Ball-and-stick Space-filling Chemical
(figure 2.3). The resulting product is called a salt. A common type a shared pair model model formula
of salt, sodium chloride (table salt), is composed of Na+ (sodium of electrons.
cations) and Cl- (chloride anions) and forms a solid crystalline
(b)
structure. Water often breaks ionic bonds because of reasons
described later. In aqueous solution, salts are electrolytes, meaning FIGURE 2.4 Covalent Bonds Covalent bonds are formed when
they conduct electricity. atoms share electrons. (a) Methane is formed when a carbon atom
fills its outer electron shell by sharing eight electrons—four belong to
H atoms and four belong to the carbon atom. (b) Different ways of
MicroByte
depicting the methane molecule.
Electrical charges from the heart are conducted by electrolytes and
can be detected on the body surface as an electrocardiogram (ECG). ? In terms of its bonding properties, why is carbon such an
important element in biological systems?
 Part I Life and Death of Microorganisms 21

The Relative Electronegativities of

TABLE 2.2 Some Atoms Common in Biology* δ–
δ+

Hydrogen 2.1 δ+
δ–
Carbon 2.5
δ+
Nitrogen 3.0 δ+
Oxygen 3.5
Hydrogen Increasing
*Electronegativity is the ability of an atom to attract electrons to itself in a bond electron density
molecule; an atom with a significantly higher electronegativity is better able δ–
to attract electrons.

Decreasing
δ+ δ+
electron density
TABLE 2.3 Non-Polar and Polar Covalent Bonds
Water molecule
Type of
Covalent Atoms Involved and Charge FIGURE 2.6 Hydrogen Bond Formation Hydrogen bonds form
Bond Distribution between water molecules because the electron-rich oxygen atom
attracts electron-poor hydrogen atoms.
Non-polar C–C C and H have similar attractions for
electrons, so there is a nearly equal
? Explain why two identical atoms joined by a covalent bond cannot
C–H form a hydrogen bond.
charge on each atom.
H–H resulting in a polar covalent bond. The slight separation of
charge is called a dipole, and is indicated by the Greek symbol
Polar O–H The O and N atoms have a stronger
attraction for electrons than do C
delta (δ); the atom that has a slight positive charge is δ+ and the
N–H atom with the slight negative charge is δ-. An example is the
and H, so the O and N have a slight
O–C negative charge; the C and H have O—H bonds in water; the oxygen atom is more electronegative
N–C a slight positive charge. than the hydrogen atom (figure 2.5). Consequently, the oxygen
atom pulls the electrons toward it, giving it a slight negative
charge and leaving each of the two hydrogen atoms with a slight
life, so cells use protein catalysts called enzymes that help break positive charge. Polar covalent bonds play a key role in biological
covalent bonds at lower temperatures. Enzymes and their functions systems because hydrogen bonds often result.
are covered in chapter 6. enzymes, p. 135
Electrons in a covalent bond may or may not be equally Hydrogen Bonds
shared between the two atoms, depending on the relative electro-
Hydrogen bonds are weak bonds formed when a hydrogen atom
negativities of the atoms; electronegativity is the ability of an
in a polar molecule is attracted to an electronegative atom in the
atom to attract electrons to itself in a molecule (table 2.2). A
same or another polar molecule (figure 2.6; see also table 2.2).
non-polar covalent bond forms when the electrons are shared
Compounds that contain electronegative atoms such as oxygen (O)
equally, such as when identical atoms share electrons. The same
or nitrogen (N) are common in biological systems, creating the
can occur between different atoms, if both have a similar attrac-
possibility for many hydrogen bonds. Like other weak bonds, these
tion for electrons (table 2.3). Examples of non-polar covalent
are important in molecule-molecule recognition (figure 2.7). For
bonds are H–H and C–H.
If one atom in a covalent bond is significantly more electro-
negative than the other, then the electrons are shared unequally, Weak bond

+ –
+
Increasing –
δ– –
δ+ electron density +
O H –
+
–
H δ+ Decreasing +
electron density

(a) (b) Molecule 1 Molecule 2

FIGURE 2.5 Polar Covalent Bonds Electrons move closer to the FIGURE 2.7 Weak Bonds and Molecular Recognition Weak
more electronegative atom in a compound, creating a polar molecule. bonds, such as ionic and hydrogen bonds, are important for molecules
(a) Lewis diagram of a water molecule. (b) Electron density model of to recognize each other. Many weak bonds are required to hold the
a water molecule. The symbol δ indicates a partial charge. two molecules together.
? Why is the oxygen atom in a water molecule more electron-rich ? Why would it be important for certain molecules to be held
than the hydrogen atoms? together by hydrogen bonds instead of covalent bonds?
22 Chapter 2 The Molecules of Life

example, in order for an enzyme to

Water molecule
break covalent bonds of a compound
(the substrate), the enzyme binds to
the substrate through many weak δ–
non-covalent bonds, such as
hydrogen and ionic bonds. δ+
A single hydrogen bond δ+
often exists for only a fraction of
a second, and enzymes are not
needed to form or break them. The Ice
hydrogen bonds between water mol-
ecules are constantly being formed and
broken at room temperature because the
energy produced by the movement of water is enough
to break the bonds.
Although a single hydrogen bond is too weak to keep mol- Liquid water
ecules together, a large number can hold them together firmly.
An analogy would be the hook and loop fasteners of Velcro. A
single hook-and-loop attachment does not provide much
strength, but many such attachments result in a strong connec-
tion. Like hook and loop attachments, weak bonds can be
formed and broken quickly and easily, allowing the molecules
to separate. A good example is the double-stranded DNA mol-
ecule. The two strands are held together by many hydrogen
FIGURE 2.8 Water In ice, each H2O molecule hydrogen bonds to
bonds along the molecule. The two strands will come apart if four other H2O molecules, forming a rigid crystalline structure. In
energy is supplied, usually in the form of heat approaching tem- liquid water, the hydrogen bonds continuously break and re-form
peratures of 100°C. and the molecules can move closer together.
? Why does water expand as it freezes?

Molarity
A chemical reaction can be likened to a recipe—it uses relative
2.3 ■ Chemical Components
quantities of different substances. But while chefs work with mea- of the Cell
sures such as a dozen, chemists work with moles. One mole is
6.022×1023 particles. That number is not important from a practi- Learning Outcomes
cal standpoint, but the concept is essential in chemistry—a mole 5. Describe the bonding properties of water molecules, and explain
of one compound has the same number of molecules as a mole of why they are important in biology.
any other. 6. Explain the concept of pH, and how the pH of a solution relates to
One mole of sodium chloride (NaCl), for example, weighs its acidity.
58.4 grams, whereas one mole of potassium chloride (KCl) 7. Name the four types of macromolecules found in all cells.
weighs 74.55 grams. The molarity (M) of a solution is defined as
the number of moles of a compound dissolved in 1 liter of solu-
tion. Therefore, a 1 M solution of NaCl has 58.4 grams of NaCl
Water
dissolved in 1 liter of aqueous solution. The most important molecule in the cell is water and the life of all
organisms depends on it. In addition, cells contain many elements,
small molecules, and macromolecules (large molecules). Water
MicroAssessment 2.2 makes up over 70% of all living organisms by weight. The impor-
tance of water in large part depends on its unusual properties.
Valence electrons are important in chemical bond formation. Ionic
bonds form between positively and negatively charged ions. Covalent
bonds result from sharing electrons. Hydrogen bonds form between Bonding Properties of Water
polar molecules or portions of molecules. Hydrogen bonds play a critical role in the properties of water.
4. Compare the relative strengths of covalent, hydrogen, and ionic Because water is a polar molecule, hydrogen bonds form between
bonds. the δ- O portions of one molecule and the δ+ H portions of
5. Which type of bond requires an enzyme to break it? another. The extent and stability of hydrogen bonding between
6. Why does an atom that loses electrons become positively water molecules depends on the temperature (figure 2.8). At
charged? What causes the positive charge? + freezing temperatures, the water molecules form a lattice-like
crystalline structure (ice). At room temperature, however, the
 Part I Life and Death of Microorganisms 23

hydrogen bonds continually break and reform. Because of the H+ ion concentration pH
fluctuations in hydrogen bonding at room temperature, the water (molarity)
molecules can move closer together, so liquid water is denser than 10-14 14 1 M NaOH
ice. This is why ice floats. Drain cleaner
The polar nature of water molecules also accounts for water’s
13 Lye
ability to dissolve a large number of compounds. To dissolve in
water, compounds must be polar or have a positive or negative

More basic (higher pH)

12
charge. The polar water molecules surround these compounds. In Household ammonia
the case of salts, water molecules split them into their component
ions. For example, NaCl dissolves in water to form Na+ and Cl-; 11
the surrounding water molecules prevent them from associating Milk of magnesia
(figure 2.9). Salts and polar molecules are hydrophilic (“water 10
loving”). In contrast, non-polar molecules are hydrophobic Detergent solution
(“water-fearing”); they do not dissolve in water because they can- 9
not form hydrogen bonds.
Water containing dissolved substances freezes at a lower 8 Seawater
temperature than pure water, so most water does not freeze Blood
unless the temperature drops below 0°C. Consequently, some 10-7 7 NEUTRAL
microorganisms can multiply below 0°C, because at least some of
the water remains liquid. Milk
Urine

More acidic (lower pH)

6
Unpolluted rainwater
pH of Aqueous Solutions
An important property of aqueous solutions is their pH, a measure of 5 Black coffee
their acidity. It is measured on a logarithmic scale of 0 to 14 in which Beer
the lower the number, the more acidic the solution (figure 2.10). 4
pH is a measure of the concentration of H+ in moles per liter.
Vinegar
Water has a slight tendency to split (ionize) into hydrogen ions 3
Cola
H+ (protons), which are acidic, and OH- ions (hydroxyl), which are
Lemon juice
2
Stomach acid
1
Water molecules δ–
δ– 100 0 Battery acid
Na+
δ– δ–
δ–
FIGURE 2.10 pH Scale The concentration of H+ ions varies by a
+ factor of 10 between each pH number since the scale is logarithmic.
Na
? Does the H+ concentration increase or decrease when the pH drops
from 5 to 4? What about the OH– concentration?
Cl–
basic or alkaline. In pure water, the concentration of H+ and OH-
ions is equal, and the concentration of each is 10-7 molar (10-7 M).
δ+ The product of the concentration of H+ and OH- is always equal to
δ+
δ+
10-14 M (10-7×10-7) (exponents are added when numbers are
Cl–
multiplied). Thus, if H+ ions are added to an aqueous solution,
δ+ increasing the concentration of H+ 10-fold (to 10-6 M), then the
δ+
concentration of OH- must decrease by a factor of 10 (to 10-8 M).
The pH scale ranges from 0 to 14 because the concentrations
Salt crystal (NaCl) of H+ and OH- ions vary within these limits. When the concentra-
tions of H+ and OH- are equal (10-7), the pH of the solution is 7.0
and neutral. For every unit on the log scale, however, the concen-
FIGURE 2.9 Salt (NaCl) Crystal Dissolving in Water In water,
the Na+ and Cl– are separated by H2O molecules. The Na+ is attracted tration of H+ ions changes by a factor of 10.
to the slightly negatively charged O– and the Cl– is attracted to the Compounds called buffers stabilize the pH of solutions. They
slightly positively charged H+ portion of the water molecules. In the are frequently added to bacterial growth media to prevent a dramatic
absence of water, the salt is highly structured because of ionic bonds rise or fall in pH resulting from metabolic processes. This is
between Na+ and Cl– ions.
important to do because most bacteria can live only within a nar-
? If water were not polar, would it dissolve sodium chloride? Explain. row pH range, usually near neutrality.
24 Chapter 2 The Molecules of Life

Elements and Small Molecules Adenosine

in the Cell NH2

All cells contain a variety of elements and small molecules, many of Phosphate groups N
N
which are ions. About 1% of a bacterial cell’s dry weight (weight O O O
excluding water) is composed of inorganic ions, principally Na+
O– P O P O P O CH2 N N
(sodium), K+ (potassium), Mg2+ (magnesium), Ca2+ (calcium), Fe2+
Adenine
(iron), Cl- (chloride), PO43- (phosphate), and SO42- (sulfate). O– O– O– O
Certain enzymes require positively charged ions in very small
amounts to function. High-energy
Organic compounds often have distinctive chemical groups bonds OH OH
that contribute to the molecule’s properties. Characteristics of Ribose
some of these functional groups are shown in table 2.4. FIGURE 2.11 ATP Adenosine triphosphate (ATP) serves as the
An especially important small organic molecule is adenosine energy currency of a cell. When the terminal phosphate bonds break,
triphosphate (ATP), the energy currency of a cell (figure 2.11). the energy released can be used to drive cellular reactions.
ATP is an effective energy carrier because the three negatively ? Why are the bonds between the phosphate groups of ATP “high
charged phosphate groups repel each other, so the bonds joining them energy”?
are inherently unstable. They are easily broken to release a sufficient
amount of energy to drive a cellular process. The relatively high When the terminal phosphate bond of ATP breaks, inorganic phos-
amount of energy released when the bonds are hydrolyzed makes phate and adenosine diphosphate (ADP) are formed. The role of
them high-energy phosphate bonds, indicated by the symbol ~. ATP in energy metabolism is covered more fully in chapter 6.
Other organic small molecules in cells are the subunits, or
Biologically Important building blocks, of macromolecules that will be described next.
TABLE 2.4 Functional Groups The subunits include amino acids, nucleotides, and various sugars.
Functional Biological
Group Structure Significance
Macromolecules and Their
Component Parts
O
Aldehyde Carbohydrates Macromolecules are large molecules (macro means “large”). The
C H four major classes are proteins, carbohydrates, nucleic acids, and
H
lipids. Although these groups differ from each other in their
Amino acids, the subunit chemical structure, they have some features in common.
Amino N
of protein Most macromolecules are polymers (poly means “many”),
H
formed by joining subunits together. Different classes of macro-
O molecules are composed of different subunits, each with distinct
Organic acids, including
Carboxyl C
amino acids and fatty acids
structures. Macromolecules are synthesized by joining subunits,
OH one by one, generally forming a long chain. This joining involves
Carbohydrates, fatty acids,
dehydration synthesis—a chemical reaction that removes H2O
Hydroxyl OH (figure 2.12). The reverse reaction, breaking a macromolecule
alcohol, some amino acids
down to its subunits by adding H2O is called a hydrolytic reaction,
O
Carbohydrates, or hydrolysis (figure 2.12b). Both types of chemical reactions
Keto
C polypeptides
require specific enzymes.
H
Some amino acids, MicroAssessment 2.3
Methyl C H
attached to DNA The weak polar bonds between water molecules are responsible for the
H many properties of water required for life. The degree of acidity of an
aqueous solution is expressed as pH. Macromolecules consist of many
O–
Nucleotides (subunit repeating subunits, joined together in a reaction that releases water.
Phosphate O P O– of nucleic acids), ATP, 7. Why is water a polar molecule? Give two examples of why this
signaling molecules property is important in microbiology.
O
8. Name the four important classes of large molecules in cells.
Part of the amino acid 9. In pure water, what must be done to decrease the OH-
Sulfhydryl S H
cysteine concentration? To decrease the H+ concentration? +
 Part I Life and Death of Microorganisms 25

■ Cell framework. Proteins make up the cytoskeleton, the struc-

Dehydration Synthesis tural framework of many cells. cytoskeleton, p. 73
■ Sensing and responding to conditions outside the cell.
H2O
Proteins on the cell surface recognize conditions in the external
HO H + HO H
environment and relay that information to the cellular machin-
ery. two-component system, p. 178
■ Regulating gene expression. Proteins bind to DNA and regu-
late gene expression. gene expression, pp. 162, 168
HO H

(a)
MicroByte
The genetic information of a typical bacterial cell encodes up to
Hydrolysis 4,000 proteins.

H2O

HO H
Amino Acids
Amino acids are the subunits that make up proteins. Twenty major
amino acids can be arranged in a functionally infinite number of
HO H + HO H
combinations in a protein. The characteristics of a protein depend
mainly on its shape, which in turn depends on the sequence of
(b) amino acids.
FIGURE 2.12 Synthesis and Breakdown of Macromolecules All amino acids have a central carbon atom bonded to (1) a
(a) Subunits are joined together (polymerized) by removing water, a carboxyl group, (2) an amino group, and (3) a side chain (R
dehydration reaction. (b) In the reverse reaction, hydrolysis, the addi- group) (figure 2.13). The side chain—the part that distinguishes
tion of water breaks bonds between the subunits. different amino acids—gives an amino acid its characteristic
? What are the four major classes of macromolecules? properties.
Amino acids are subdivided into several different groups
based on properties of their side chains (figure 2.14). Non-polar
amino acids are characterized by side chains that lack polar
bonds; an example is the methyl group (–CH3) of alanine. In
2.4 ■ Proteins contrast, polar amino acids have side chains that contain a polar
bond; an example is the hydroxyl group (–OH) of serine.
Learning Outcomes Charged amino acids carry a positive or negative charge because
8. Describe the important functions of proteins in cells. their side chains contain functional groups that can ionize; these
9. Describe the characteristics of amino acids and the peptide bonds include carboxyl groups (–COOH; acidic) and amino groups
that hold them together. (–NH2; basic).
10. Compare and contrast the four levels of protein structure.
11. Describe protein domains, substituted proteins, and protein
denaturation.
Side chain—
Proteins make up more than half of the dry weight of cells. Some “R” is the general
of their most important roles include: R designation for a side chain

H
■ Catalyzing reactions. Enzymes are proteins that speed up the O
Amino group— Carboxyl group—
various chemical reactions in cells. enzymes, p. 135 H N+ C C
positively charged negatively charged
at neutral pH H H O– at neutral pH
■ Transporting molecules. Transport proteins move
molecules either into or out of cells. transport proteins, FIGURE 2.13 Generalized Amino Acid This figure illustrates the
p. 55 three groups that all amino acids possess.
■ Motility. Proteins are essential components of flagella and ? Which portion of an amino acid is responsible for the unique
cilia, structures that move cells. flagella, pp. 63, 74 properties of the molecule?
26 Chapter 2 The Molecules of Life

Non-polar

H3C CH3 CH3

H3C CH3 CH CH2 CH2
H CH3 CH CH2 H3C CH H2C CH2
H3 N+ C COO– H3 N+ C COO – H3 N+ C COO –
H3N+ C COO –
H3N+ C COO – H2N+ C COO–
H H H H H H
Glycine (Gly; G) Alanine (Ala; A) Valine (Val; V) Leucine (Leu; L) Isoleucine (IIe; I) Proline (Pro; P)

H CH3
N S
SH CH2
CH2 CH2 CH2 CH2
H3N+ C COO– H3N+ C COO– H3N+ C COO– H3N+ C COO–
H H H H
Phenylalanine (Phe; F) Tryptophan (Trp; W) Cysteine (Cys; C) Methionine (Met; M)

Polar/hydrophilic (uncharged)

OH
O NH2
O NH2 C
OH HO CH3 C CH2
CH2 CH CH2 CH2 CH2
H3N+ C COO– H3N+ C COO– H3N+ C COO– H3N+ C COO– H3N+ C COO–
H H H H H
Serine (Ser; S) Threonine (Thr; T) Asparagine (Asn; N) Glutamine (Gln; Q) Tyrosine (Tyr; Y)

Polar/hydrophilic (charged)

Basic

Acidic NH2
NH3 + C NH2+

O O– CH2 NH

O O– C HN CH2 CH2
NH+
C CH2 CH2 CH2
CH2 CH2 CH2 CH2 CH2
H3 N+ C COO – H3N+ C COO –
H3N+ C COO –
H3 N+ C COO – H3 N+ C COO–
H H H H H
Aspartic acid (Asp; D) Glutamic acid (Glu; E) Histidine (His; H) Lysine (Lys; K) Arginine (Arg; R)

FIGURE 2.14 Common Amino Acids The groupings are based on the polarity and the overall charge of the amino acid. The basic and acidic
amino acids have a net positive and negative charge, respectively. For simplicity, tyrosine is shown in only one group but it has both non-polar and
polar characteristics. The three-letter and single-letter code names for each amino acid are given.
? What chemical groups characterize a hydrophobic amino acid? A hydrophilic amino acid?
 Part I Life and Death of Microorganisms 27

L-Amino acid D-Amino acid consists of about 250 amino acids. The primary structure in large
part determines the final shape of the protein and thus is respon-
W W
sible for its properties.
The secondary structure is the three-dimensional shape of
C C localized regions; certain amino acid sequences (the primary struc-
ture) lead to characteristic spirals and folds due to weak forces
X Y Z Z Y X such as hydrogen bonds (figure 2.17b). A spiral or helical struc-
ture is an alpha (α) helix, whereas parallel strands make up a beta
(β) pleated sheet.
The entire protein folds into its distinctive three-dimensional
shape, its tertiary structure (figure 2.17c). The tertiary structure
is determined primarily by the sequence of amino acids and
whether or not they interact with water. Amino acids that have
polar side chains are hydrophilic, and are typically located on the
Mirror outside of the protein molecule, where they can interact with
charged polar water molecules. Amino acids that have non-polar
side chains are hydrophobic, so these tend to cluster inside the
FIGURE 2.15 Mirror Images (Stereoisomers) of an Amino Acid protein molecule, thereby avoiding water molecules. In addition to
The joining of a carbon atom to four different groups leads to asym- these interactions, a covalent bond can form between sulfur atoms
metry in the amino acid. The molecule can exist in either the L or D in different cysteine molecules, creating a disulfide (S–S) bond.
form, each being the mirror image of the other. The two molecules
cannot be rotated in space to give two identical molecules.
The combination of strong and weak bonds between the various
amino acids results in the proteins’ tertiary structure. Two major
? Which form (L or D) is found in proteins?
shapes exist: globular, which tend to be spherical and water solu-
ble, and fibrous, which are elongated and insoluble.
Proteins sometimes consist of more than one polypeptide,
All amino acids except glycine can exist in two stereoisomers, either identical or different from one another, which are held
or spatial arrangements (figure 2.15; see A Glimpse of History). together by many weak bonds. The specific shape that results is
These forms—L (left-handed) and D (right-handed)—are mirror the quaternary structure (figure 2.17d). Of course, only proteins
images of each other. Proteins only have L-amino acids. The D-amino that consist of more than one polypeptide have a quaternary
acids are found in a few structures in bacteria, but otherwise are rare
in nature.

R R
Peptide Bonds
H H
Amino acids are held together in an unbranched chain by peptide O O
bonds. This type of covalent bond forms when the carboxyl group H N+ C C + H N+ C C
of one amino acid reacts with the amino group of another, releasing
O– O–
water (dehydration synthesis) (figure 2.16). The joining of amino H H H H
acid subunits by peptide bonds creates a polypeptide. One end of the Amino acid Amino acid
molecule has a free amino group (the N terminal or amino terminal
end), and the other has a free carboxyl group (the C terminal or car- Dehydration
boxyl terminal end). protein synthesis, p. 170 H2O synthesis
A protein is one or more long polypeptides folded to create a
functional molecule. Note, however, that the distinction between a R R
polypeptide and a protein is not always clear, and the terms are often H O
used interchangeably. O
H N+ C C N C C
O–
H H H H
Protein Structure
Proteins have four levels of structure: primary, secondary, tertiary, FIGURE 2.16 Peptide Bond Formation Dehydration synthesis
and quaternary. The number and sequence of amino acids in the forms the peptide bond, shown in red.
polypeptide determines its primary structure (figure  2.17). ? What two chemical groups are involved in the formation of a
Proteins vary greatly in size, but an average-size polypeptide peptide bond?
28 Chapter 2 The Molecules of Life

FIGURE 2.17 Protein Structure (a) The primary structure is deter-

Primary Structure mined by the amino acid composition. (b) The secondary structure
results from folding of the various parts of the protein into two major
patterns—helices and sheets. For simplicity, R groups are not shown.
R R (c) The tertiary structure is the overall shape of the molecule. The dia-
H H O H H
gram represents a ribbon model of the protein, made up of helices and
C C N C C N C C N C sheets. (d) The quaternary structure results from several polypeptide
chains interacting to form the protein. The molecule shown in (c) can
H O R H H O R weakly bond with an identical protein molecule to form a dimer.
? Which of the four levels of structure are especially important in
The primary structure can fold determining the properties of a protein?
into a pleated sheet, or turn into a helix.

(a)

Secondary Structure Secondary Structure

β-pleated sheet α-helix

(b)

Tertiary Structure Quaternary Structure

(c) (d)

structure. The individual polypeptides generally do not have bio- Protein Domains
logical activity.
A unit of organization distinct from the four levels just described
After being synthesized, the polypeptide chain folds into its
is the protein domain (figure 2.18). A domain is a protein sub-
correct shape. Although many shapes are possible, only one is
structure consisting of sheets and helices that fold into a stable
functional. Most proteins fold spontaneously into their correct
structure independently of other parts of the molecule. Large pro-
state, but protein chaperones are sometimes needed to help with
teins sometimes contain many domains, whereas a small protein
the process. Misfolded proteins are degraded into their amino acid
may contain only one. The domains, usually consisting of 40 to
subunits, which are then used to make more proteins.
350 amino acids, are connected to each other by short lengths of
MicroByte polypeptides.
The computer game “Foldit” allows players to help researchers
Different domains are associated with specific functions. For
predict how a specific amino acid sequence will fold to become a
example, a certain domain may grasp DNA; another may carry out
functional protein.
a catalytic activity. Thus, a single protein may have several different
 Part I Life and Death of Microorganisms 29

Protein Denaturation
High temperature, extreme pH, and certain solvents can break
Domain 1 bonds within a protein, causing its shape to change (figure 2.19).
The protein becomes denatured and no longer functions. This is
why most organisms cannot grow at very high temperatures—
their enzymes denature. Denaturation may be reversible in some,
but not all, cases. If the denaturating agent is a chemical that is
then removed, for example, the protein may refold spontaneously
into its original shape. However, boiling an egg denatures the egg
white protein irreversibly, which is why cooling the egg does not
restore it to its original appearance.

MicroAssessment 2.4
Domain 3 The properties of amino acids are determined by their side chains.
The sequence of amino acids in a protein determines how the protein
Domain 2 folds into a functional three-dimensional shape. Substituted proteins
have other molecules attached.
FIGURE 2.18 Domain Structure of a Protein Each domain has 10. What type of bond joins amino acids to form proteins?
different functions.
11. Describe five roles of proteins.
? Which levels of protein structure determine the properties of 12. What elements must all amino acids contain? What element will
domains? only some amino acids contain? +

functions. Once a known function can be attributed to a specific

domain, the function of an unknown protein can be inferred if it has 2.5 ■ Carbohydrates
that domain.
Learning Outcomes
Substituted Proteins 12. Describe the important functions of carbohydrates in a cell.
Substituted proteins have other molecules covalently bonded to the 13. Compare and contrast monosaccharides, disaccharides, and
side chains of some of their amino acids. Many proteins found on polysaccharides.
the surface of cells are substituted. The proteins are named after the
molecules covalently joined to the amino acids. If sugar molecules Carbohydrates are a diverse group of compounds that include
are bonded, the protein is a glycoprotein; if lipids are attached, the sugars and starches. They play several critical roles in biology:
protein is a lipoprotein. Sugars ■ Energy source. Organisms degrade carbohydrates to harvest
and lipids are covered later
the energy they contain. metabolism, p. 126
in this chapter.
■ Energy storage. Organisms can store excess energy and
nutrients for later use by producing certain carbohydrates
that function as reserve material. storage granules, p. 66
■ Source of carbon for biosynthetic products. Many
microbes can make all of their cell components from a sin-
gle carbohydrate—glucose. precursor metabolites, p. 132
Heated to 100°C ■ Component of DNA and RNA. The subunits of
DNA and RNA contain sugars. nucleic acids, p. 32
■ Structural components of cells. Some types of
Properly folded protein (active)
cell walls are composed of sugar-containing material.
cell wall structure, p. 58

Carbohydrates all contain carbon, hydrogen,

and oxygen atoms in an approximate ratio of 1 : 2 :1.
Their general chemical formula, (CH2O)n, indicates
Denatured protein (inactive) this ratio. The “H2O” in that formula is reflected in the
FIGURE 2.19 Denaturation of a Protein The denatured term “carbohydrate” (meaning “hydrate of carbon”); note,
protein loses its function. however, that the arrangement of atoms in carbohydrate
? Describe two environmental conditions that can denature a protein. molecules has little in common with water.
30 Chapter 2 The Molecules of Life

Monosaccharides H
5CH2OH
O
Monosaccharides, or simple sugars, have only a single unit 1C O OH
(mono means “one”). They are classified by the number of car- 4C H H C1
H 2
C OH
bon atoms they contain, and most common monosaccharides H C3 C2 H
have five or six carbon atoms (table 2.5). Each carbon atom is H 3
C OH
OH OH
numbered using a characteristic scheme, allowing scientists to
H C OH Ribose
describe the position of various functional groups attached to the 4

molecules (figure 2.20). H C OH

5

Common Monosaccharides, H
TABLE 2.5 5CH2OH
Disaccharides, and Polysaccharides O
1C O OH
Name Components Significance 4C
H C H H H C1
2 H C3
Monosaccharides C2 H
H 3
C OH
(5-carbon) OH H
Ribose Component of RNA H C OH Deoxyribose
4

Deoxyribose Component of DNA H C OH

5
(6-carbon)
H
Glucose Common subunit of
disaccharides FIGURE 2.20 Ribose and Deoxyribose Linear and ring forms.
Galactose Component of milk Although both structures occur in the cell, the ring form predomi-
sugar (see below) nates. The plane of the ring is perpendicular to the plane of the paper
with the thick line on the ring closest to the reader.
Fructose Fruit sugar
? What is the major chemical difference between ribose and
Mannose Found on the surface deoxyribose?
of some microbes
6CH2 OH 6CH2 OH
Disaccharides 5 O 5 O
H H H H H H
Lactose Glucose + Milk sugar 4 1 4 1
galactose OH H OH OH
HO OH HO OH
3 2 3 2
Maltose Glucose + Breakdown product
glucose of starch H OH H H
Glucose Mannose
Sucrose Glucose + Table sugar from
fructose sugar cane and beets 6CH2 OH 6CH2 OH
5 O O
Polysaccharides HO H H OH
4 1
Agar Polymer of Gelling agent in OH H 5 H OH 2

galactose bacteriological H OH H 1CH2 OH

3 2 4 3
media; extracted H OH OH H
from the cell walls Galactose Fructose
of some algae
FIGURE 2.21 Common 6-Carbon Sugars These sugars are struc-
Cellulose Polymer of Main structural tural isomers with different properties.
glucose, in a polysaccharide in
β 1,4 linkage; plant cell walls ? What is a structural isomer?
no branching
Chitin Polymer of Major organic Ribose
N-acetyl- component in
glucosamine exoskeleton of insects 5CH2OH 5CH2OH
and crustaceans O O
H OH
Dextran Polymer of Storage product in 4
H H 1 4
H H 1
glucose in an some bacterial cells H OH H H
3 2 3 2
α 1,6 linkage;
branching OH OH OH OH
Glycogen Polymer of Main storage α form β form
glucose in an polysaccharide in
α 1,4 linkage; animal and bacterial
branching cells FIGURE 2.22 α and β Forms The α and β forms of ribose are
interconvertible and differ only in whether the OH group on carbon 1
Starch Polymer of Main storage product is above or below the plane of the ring.
glucose in plants
? When are the α and β forms not interconvertible?
 Part I Life and Death of Microorganisms 31

Sugars occur in two interchangeable CH2OH CH2OH H 2O

CH2OH CH2OH
forms: linear and ring (figure 2.20). Both O O O O
H H H H H H H H
naturally occur in the cell, but most are in OH H
+
H OH OH H H OH
the ring form. In diagrams, the lower por- HO OH HO CH2OH Dehydration HO O CH2OH
tion of the ring form is thickened to suggest H OH OH H synthesis H OH OH H
a three-dimensional structure. α--Glucose Fructose Sucrose
Ribose and deoxyribose are 5-carbon FIGURE 2.23 Formation of a Disaccharide The sucrose molecule is formed by the removal
sugars found in nucleic acids (figure 2.20). of water.
These sugars are identical except deoxyri- ? What type of reaction would reverse the step shown in this diagram?
bose has one less molecule of oxygen than
does ribose (de means “away from”); ribose
has a hydroxyl group on the number 2 car- Cellulose
bon (also called the 2 prime carbon, written 2 carbon), whereas
deoxyribose has only a hydrogen at that position.
Glucose, galactose, fructose, and mannose are all
Weak bond
6-carbon sugars. These are all structural isomers, meaning that
they contain the same atoms but differ in their chemical
arrangements (figure  2.21). Structural isomers result in dis-
6CH2OH 6
CH2OH
tinct sugars with different properties and different names. For
example, both glucose and mannose have a sweet taste, but 5C O 5C O
mannose has a bitter aftertaste. Glucose, an important energy O 4C C1 O C4 C1 O
Non-branching
source of many cells, will be discussed extensively in the
chapter on metabolism. Mannose is found on the surface of 3C C2 3C C2
Glucose
some microbes.
Sugars can exist in two different forms—alpha (α) and
beta (β)—based on the relative position of the hydroxyl group Glycogen
joined to the number 1 carbon atom (figure 2.22). The α and
β forms are interconvertible, but once the carbon atom is
6CH2OH 6
CH2OH
joined to another sugar molecule, the α or β form is essentially
locked in place. 5C O 5C O

4C C1 C4 C1
O O O
Disaccharides 3C C2 3C C2

Disaccharides are two monosaccharides joined together by

covalent bonds (table 2.5). The two most common examples are
sucrose (table sugar) and lactose (milk sugar). Sucrose, which
Branching
comes from sugar cane and sugar beets, is composed of glucose
and fructose, whereas lactose consists of glucose and galactose.
Another disaccharide, maltose, composed of two glucose mol- Dextran
ecules, is a breakdown product of starch.
To form a disaccharide, two monosaccharides are joined
6CH2
together by a dehydration reaction between a pair of their
5C O
hydroxyl groups, with the loss of water (figure 2.23). Note that
this reaction is similar to that used to join two amino acids. The 4C C1 6
reaction is reversible, so hydrolysis, which adds a water mole- O CH2
3C C2
cule, yields the two original molecules. 5C O

4
C C1
O
3C C2
Polysaccharides
Polysaccharides are large molecules composed of long chains Branching
of monosaccharide subunits or their derivatives (table 2.5).
Shorter chains are called oligosaccharides. FIGURE 2.24 Structures of Three Important Polysaccharides
Polysaccharides often contain only glucose molecules, but The molecules shown have the same subunit (glucose) yet they are
are structurally diverse because some polymers are branched. In distinct because of differences in linkage that join the molecules
addition, some types have linkages between α forms of the sug- (α and β; 1,4 or 1,6), the degree of branching, and the bonds involved
in branching (not shown). Weak bonding forces are also involved.
ars and others between β forms. The position of the carbon
atoms involved in the bonding can also differ (figure 2.24). ? Where are the three polysaccharides shown found in nature?
32 Chapter 2 The Molecules of Life

Cellulose, starch, glycogen, and dextran, all polymers of Adenine

NH2
glucose, are important polysaccharides. Cellulose is the princi- N
N
ple component of plant cell walls and the most abundant organic H Nucleobase
molecule on earth. Most organisms cannot degrade this sub- N H
N
stance, however, because they lack the enzyme that  breaks the O–
bonds joining the subunits. Certain bacteria and fungi have that
HO P O 5CH2
enzyme, which is why they play such an important role in recy- O
cling organic material. Starch, the energy storage form produced OH Deoxyribose
4 1
by plants, can be used as a food source by many organisms. H H (5-carbon sugar)
H H
Glycogen is an energy storage product of animals and some bac- Phosphate 3 2
teria. Dextran, a storage product of some microbes is a compo- group OH H
nent of some products used to increase the volume of blood or
deliver iron to the blood of iron-deficient patients. cellulose FIGURE 2.25 A Nucleotide This is one subunit of DNA. This
degradation, p. 150
subunit is called adenylic acid or deoxyadenosine-5-phosphate
because the nucleobase is adenine. If the nucleobase is thymine,
Chitin and agar are other important polysaccharides in the nucleotide is thymidylic acid; if guanine, guanylic acid; and if
microbiology. Chitin, a polymer of the glucose derivative cytosine, cytidylic acid. If the nucleotide lacks the phosphate group,
N-acetylglucosamine, is in the cell walls of fungi and is a major it is called a nucleoside—in this case, deoxyadenosine.
component in the exoskeletons of insects and crustaceans. Agar, a ? What are the three components of a nucleotide?
polymer of galactose, is found in the cell walls of algae; it is exten-
sively used as a gelling agent in media used to grow microorgan-
isms in the laboratory. of two fused rings; and pyrimidines (cytosine and thymine), single
ring structures (figure 2.26).
MicroAssessment 2.5 Nucleic acids consist of linear chains of nucleotide subunits
with a covalent bond between the phosphate of one nucleotide and
Carbohydrates all have the general formula (CH2O)n. Sugars can
exist in a number of different isomeric forms that have different the sugar of the next (figure 2.27). Thus, the phosphate is a bridge
properties. Monosaccharides are the subunits of disaccharides and that joins the number 3 carbon (3, pronounced “3 prime”) of one
polysaccharides. Polysaccharides consisting of the same sugars sugar to the number 5 carbon (5) of the other. This results in a
have different properties because of differences in bonding between molecule with a backbone of alternating sugar and phosphate mol-
subunits. ecules. The 5 end of the chain has a phosphate attached to the
13. Distinguish between structural isomers and stereoisomers. number 5 carbon of sugar; the 3 end has a hydroxyl group
14. What is the general name given to a single sugar? How can attached to the number 3 carbon (figure 2.27). During DNA syn-
single sugars differ from another? thesis, the chain is elongated by adding more nucleotides to the
15. How can you distinguish sucrose and lactose from a protein hydroxyl group at the 3 end. This topic is covered in chapter 7.
molecule by identifying the elements in the molecules? +

Purines Pyrimidines
2.6 ■ Nucleic Acids (double ring) (single ring)
Learning Outcome NH2 O O
CH3 H H
14. Compare and contrast the chemical compositions, structures, and N N H N
N
major functions of DNA and RNA. H
N H H O H O
N N N
Nucleic acids carry genetic information. Cells decode this infor-
H H H
mation, converting the information in a sequence of nucleotides
Adenine (A) Thymine (T) Uracil (U)
into the sequence of amino acids in protein molecules. (both DNA and RNA) (DNA only) (RNA only)

O NH2
DNA N H H
N N
DNA, which stands for deoxyribonucleic acid, is the master mol- H
ecule of the cell. Its nucleotide sequence encodes all of the cell’s N NH2 H O
N N
properties. The information in DNA is converted to the form of
H H
RNA, which is then translated to make proteins, a process covered
Guanine (G) Cytosine (C)
in chapter 7. (both DNA and RNA) (both DNA and RNA)
The nucleotides of DNA have three different parts: a
nucleobase, deoxyribose, and a phosphate group (figure 2.25).
The four different nucleobases in DNA can be characterized by FIGURE 2.26 Formulas of Purines and Pyrimidines
their ring structures: purines (adenine and guanine), each consisting ? Which of the nucleobases are found in DNA? In RNA?
 Part I Life and Death of Microorganisms 33

Sugar- RNA
phosphate RNA, which stands for ribonucleic acid, is involved in the proc-
backbone Nucleobases
ess that decodes the information in DNA to create a sequence of
amino acids in proteins. This complex multistep process will be
G described in chapter 7.
5
Although the structure of RNA resembles that of DNA, sev-
P
1 eral differences should be noted. First, the nucleotides in RNA
4
5' phosphate contain the pyrimidine uracil in place of thymine and the sugar
3 2 ribose in place of deoxyribose (see figures 2.20 and 2.26). Also,
whereas DNA is a long, double-stranded helix, RNA is consider-
A
ably shorter and exists as a single chain of nucleotides. Although
P 5 single-stranded, RNA may form short, double-stranded stretches
1
4 as a result of hydrogen bonding between complementary nucleo-
bases in the single strand.
3 2
Nucleotide
MicroAssessment 2.6
C
5
DNA is a double-stranded helical molecule composed of repeating
P subunits that consist of a nucleobase, a phosphate molecule, and the
1
4 sugar deoxyribose. RNA is a single-stranded molecule of repeating
3
nucleotides that contain uracil in place of thymine and ribose in place
2
of deoxyribose. DNA carries the genetic code in the sequence of
nucleotides. The information in the code is transferred to RNA and
T then into a sequence of amino acids in proteins.
P 5 16. How do the nucleotides of DNA differ from those of RNA?
The 3' carbon of 1
one nucleotide is 4 17. How does the structure of DNA differ from that of RNA?
linked to the 5' carbon 18. If the DNA molecule were placed in boiling water, how would
3 2
of the next nucleotide
OH the molecule change? +
via a phosphate group.

3' hydroxyl
2.7 ■ Lipids
FIGURE 2.27 Single Strand of DNA Single chain of nucleotides
in DNA showing the differences between 5 end and 3 end.
Learning Outcome
? What parts of the nucleotides are joined together? 15. Compare and contrast the structure and function of simple lipids,
compound lipids, and steroids.
The DNA of a typical bacterium is a single double-stranded
helix, arranged somewhat like a spiral staircase with two railings Lipids play an indispensable role in all cells. They are critically
(figure 2.28). The railings represent the sugar-phosphate back- important in the structure of membranes, which function as a
bone of the molecule, and the stairs are pairs of nucleobases cell’s gatekeepers. Membranes prevent cell contents from leak-
attached to the railings. The two strands, each about 4 million ing out, and also keep many molecules from entering cells.
nucleotides in a typical bacterium, are antiparallel, meaning they cytoplasmic membrane, p. 52

are oriented in different directions. One goes in the 3 to the 5 Lipids are a very diverse group of non-polar, hydrophobic mol-
direction; the other, 5 to 3. Each pair of nucleobases is held ecules. Their single common feature is that they are only slightly
together by hydrogen bonds. These bonds are weak, but the large soluble in water but highly soluble in organic solvents such as ether,
number of them holds the two strands of the DNA molecule firmly benzene, and chloroform. Thus, their defining characteristic is a
together. physical, rather than a chemical, property. Unlike other macromol-
The hydrogen bonding between nucleobases is specific in that ecules, lipids are not composed of similar subunits; rather, they
adenine (A) can only bond to thymine (T), and guanine (G) to consist of a wide variety of chemically distinct substances.
cytosine (C). Three hydrogen bonds join each G to C, but only two
join A to T. The pair of nucleobases that bond are said to be Simple Lipids
complementary to each other. Thus, G is complementary to C, Simple lipids contain only carbon, hydrogen, and oxygen. The most
and A to  T. These are referred to as base-pairing rules. As a common simple lipids are fats—fatty acids linked to glycerol
result, one strand of DNA is complementary to the other, and the (figure 2.29). Fatty acids are long chains of C atoms bonded to H
sequence of one strand determines the sequence of the other. atoms, with a carboxyl group on one end (figure 2.29a). The length
of the chain varies, depending on the fatty acid, which usually has an
MicroByte
Scientists once believed that DNA could not play an important role even number of carbon atoms. Glycerol is a 3-carbon molecule with
in the cell because it has such a simple structure. a hydroxyl group attached to each carbon. Fatty acids can join to
glycerol via covalent bonds between a hydroxyl group of glycerol
34 Chapter 2 The Molecules of Life

3' end
OH
5' end C 2 3
4
G 1 P
5
5
P 1 2
4 T 3
4
3 2 A
1
5
P
P 5
1 3' 5'
4 2
3
4
3 2 A 1
5
P
P 5 T G C
1
4 2 3
2 A T
3 C 4
1 P
5 G 5 T A
P 1
4 2 3 G C
3 2 1
4
A P
5 T A
5
P 1 T
4
3 2 A T
OH 5' end
G C
3' end C G

5' 3'
2C
H 5 T A
O O–
A T
O Guanine
Cytosine O
P P
O H N H2 N H O A T
O– O O
5 CH
2 H
H G C
O N N H
N H 2
4 3
H 1 H
H N
N
H
H A T
H 1 O
3 NH2 O 4
G C
2 H
O H 2C C G
O Adenine H 5
Thymine O–
P O
O H2N
N H P T A
O– H3C O O
5 CH H O
2 N H
N 2
O H N H G C
4 H
3
H A T
N 1
H N H
H
H 1
O
H
3 O 4
A T
OH 2 H
H 2C
H 5 C G
O–
O
3' P
O
O 5'
T A
5' 3'
(a) (b)
FIGURE 2.28 DNA Double-Stranded Helix (a) The sugar-phosphate backbone and the hydrogen bonding between bases. There are two
hydrogen bonds between adenine and thymine and three between guanine and cytosine. (b) The “spiral staircase” of the sugar-phosphate back-
bone, with the nucleobases on the inside.
? Which would require a higher temperature to denature—a DNA strand composed primarily of A-T base pairs or one that is the same length
but composed primarily of G-C base pairs?

and the carboxyl group of the fatty acid (figure 2.29b). A monoglyc- more double bonds. Those that have one double bond are monoun-
eride has only one fatty acid bound to glycerol; a diglyceride has two, saturated, and those with more than one double bond are  polyun-
and a triglyceride, three. In nature, the most common fats are triglyc- saturated. Oleic acid is a common monounsaturated fatty acid
erides, which are stored in the body as an energy reserve. (figure 2.29a). Most naturally occurring fatty acids are cis, meaning
Although hundreds of different fatty acids exist, they can be the hydrogen atoms attached to the double-bonded carbon mole-
divided into two groups based on the presence of double bonds cules are on the same side of the bond. Trans fatty acids have hydro-
between carbon atoms. Saturated fatty acids have no double bonds gen atoms on opposite sides of the double bond.
(figure 2.29a). The term “saturated” means they have the maximum The type of fatty acids in a triglyceride affects the melting point
number of hydrogen atoms. Unsaturated fatty acids contain one or of the fat. Fats that contain only saturated fatty acids are typically solid
 Part I Life and Death of Microorganisms 35

Saturated fatty acid (palmitic acid)

Watery exterior of cell
O H H H H H H H H H H H H H H H
HO C C C C C C C C C C C C C C C C H
H H H H H H H H H H H H H H H
Phospholipid
bilayer 
Unsaturated fatty acid (oleic acid)
H H
H H
H C H
H C C H
H C C H
H C C H
H C H C H Watery interior of cell
H C H H C H
O C H H C H
C H H C
C H H C
C H H H
HO H H
H H
Double bond Phospholipid

Polar head R
(a)
group O
O P O–
H O H O Phosphate
O
H C OH HO C R H C O C R Hydrophilic group
3 H2O CH2 CH CH2
head
O O O O Glycerol
C O C O
H C OH + HO C R H C O C R Hydrophobic
Dehydration tail CH2 CH2
O synthesis O CH2 CH2
H C OH HO C R H C O C R CH2 CH2
H H CH2 CH2
CH2 CH2
Glycerol + 3 fatty acids Triglyceride (fat)
CH2 CH2
(b)
CH2 CH2
FIGURE 2.29 Fat Formation from Fatty Acids and Glycerol CH
CH2
(a) Two common fatty acids. Most fatty acids contain an even number
CH2 CH
of carbon atoms (commonly 16 or 18) and may be saturated or
unsaturated. The unsaturated fatty acids may be cis or trans. CH2 CH2
(b) Dehydration synthesis in the joining of fatty acids with glycerol; CH2 CH2
the R groups are carbon-hydrogen chains, such as those shown in (a). CH2
CH2
? What characteristic of the fat in this figure makes it a triglyceride? CH2 CH2
CH2
CH2
at room temperature because the straight, long tails of the fatty acids CH2
CH2
can pack tightly together. Fats that contain one or more unsaturated CH2
CH2
fatty acids tend to be liquid at room temperature because these fatty CH3
CH3
acids have kinks in their long tails that prevent tight packing. Oils are
fats that are liquid at room temperature. Manufacturing processes that Saturated fatty acid Unsaturated fatty acid
hydrogenate oils, thereby making them solid at room temperature, FIGURE 2.30 Phospholipids Are an Essential Component
sometimes convert cis fatty acids to trans fatty acids. of Cell Membranes In phospholipids, two of the –OH groups of
glycerol are linked to fatty acids and the third –OH group is linked
MicroByte to a hydrophilic head group, which contains a phosphate ion and a
Diets rich in saturated fats and trans fats raise blood cholesterol polar molecule (labeled R).
levels, leading to clogged arteries. ? What about the structure of a phospholipid makes one portion
hydrophilic and the other hydrophobic?

Compound Lipids
Compound lipids contain fatty acids and glycerol as well as ele- cell from the outside environment (figure 2.30). The phospholipid
ments other than carbon, hydrogen, and oxygen, Biologically, molecules orient themselves in the membrane as opposing layers,
some of the most important of these are phospholipids, which forming a bilayer. The fatty acids face inward, interacting with the
contain a phosphate group linked to one of a variety of other polar fatty acids of the phospholipid molecules in the opposing layer. The
molecules (figure 2.30). This phosphate-containing portion is the hydrophilic polar heads face outward, toward either the aqueous
polar head and is soluble in water (hydrophilic). In contrast, the external environment or the internal (cytoplasmic) environment.
fatty acid portion is insoluble in water (hydrophobic). Water-soluble substances cannot pass through the hydrophobic por-
Phospholipids are an essential component of cytoplasmic tion, so the cell uses special transport mechanisms to move these
membranes, the structure that separates the internal contents of a across the membrane. These will be discussed in chapter 3.
36 Chapter 2 The Molecules of Life

TABLE 2.6 Structure and Function of Macromolecules

Name Subunit Some Functions of Macromolecules

Protein Amino acid Catalysts; structural portion of many cell components

Nucleic acids
DNA Deoxyribonucleotide Carrier of genetic information
RNA Ribonucleotide Various roles in protein synthesis
Polysaccharide Monosaccharide Structural component of plant cell wall; storage products
Lipids Varies—subunits are not similar Important component of cell membranes

Steroids
Steroid Ring Sterol (cholesterol)
Steroids are simple lipids that have a characteristic structure con-
H3C
sisting of four connected rings (figure 2.31). Their chemical
CH3 CH3
C
structure is quite different from fats, but they are classified as
CH3 lipids because they are insoluble in water. If a hydroxyl group is
CH3
attached to one of the rings, the steroid is a sterol, an example
being cholesterol (figure 2.31b). Other steroids include the hor-
mones cortisone, progesterone, and testosterone.
HO
Some of the most important properties of macromolecules of
Hydroxyl group attached to a ring
biological importance are summarized in table 2.6.
(a) (b)

FIGURE 2.31 Steroid (a) General formula showing the four- MicroAssessment 2.7
membered ring and (b) the –OH group that makes the molecule a
sterol. The sterol shown here is cholesterol. The carbon atoms in the Lipids are a diverse group of hydrophobic molecules. Fats are
ring structures and the attached hydrogen atoms are not shown. composed of fatty acids joined to glycerol. Phospholipids, with one
end hydrophilic and the other hydrophobic, form a major part of cell
? Why are steroids classified as lipids? membranes where they exist as a bilayer. They limit the entry and exit
of molecules into and out of cells.
19. What are the main functions of lipids in cells?
Other compound lipids are found in bacterial membranes and
will also be discussed in chapter 3. These include the lipoproteins 20. What features in the chemical composition of phospholipids
make them ideal components of the cytoplasmic membrane?
(covalent associations of proteins and lipids) and lipopolysaccha-
rides (molecules of lipid linked with polysaccharides through 21. How could you determine if a solid compound were a lipid or a
carbohydrate based on its solubility properties? +
covalent bonds).

Summary
2.1 ■ Atoms and Elements Molarity
Atoms are composed of electrons, protons, and neutrons (figures 2.1, A mole of one compound has the same number of molecules as a mole
2.2). An element consists of a single type of atom. Valence electrons are of another compound.
particularly important in chemical bond formation. 2.3 ■ Chemical Components of the Cell
2.2 ■ Chemical Reactions and Bonds Water
Water is the most important molecule in the cell, making up over 70%
Ions and Ionic Bonds of all living organisms by weight. Hydrogen bonding plays a very
Ionic bonds are weak attractions between cations and anions (figure 2.3). important role in the properties of water (figures 2.8, 2.9). pH is the
Covalent Bonds degree of acidity of a solution and is measured on a scale of 0 to 14.
Covalent bonds are formed by atoms sharing electrons (figure 2.4). Buffers prevent a dramatic rise or fall of pH (figure 2.10).
Organic compounds have C–H bonds. When atoms have an equal Elements and Small Molecules in the Cell
attraction for electrons, a non-polar covalent bond is formed (table 2.3). All cells contain a variety of elements and small molecules. Organic mol-
When one atom is electronegative, a polar covalent bond is formed ecules often contain distinct chemical groups important in the function-
(figure 2.5). ing of the molecule (table 2.4). ATP carries energy in two high-energy
Hydrogen Bonds phosphate bonds, which, when broken, release the energy (figure 2.11).
Hydrogen bonds are weak bonds that result from the attraction of a Macromolecules and Their Component Parts
hydrogen atom in a polar molecule to an electronegative atom in another Macromolecules are usually polymers of subunits. Different classes
polar molecule (figure 2.6). Hydrogen bonds and ionic bonds are important of macromolecules have different subunits. Macromolecules are made
in the weak association of various molecules with one another (figure 2.7). through dehydration synthesis and degraded by hydrolysis (figure 2.12).
 Part I Life and Death of Microorganisms 37

FUTURE CHALLENGES 2.1

Fold Properly: Do Not Bend or Mutilate
An organism’s properties depend on its fold incorrectly, we might be able to prevent the amino acid side chains allow the flex-
proteins, including structural proteins and such diseases. ible molecule to “find its way” to the correct
enzymes. Even though a cell may be able The information that determines how a tertiary structure. Proteins called chaperones
to synthesize a protein, that protein will not protein folds into its three-dimensional shape can assist the process by preventing detri-
function properly unless it is folded correctly is contained in the sequence of its amino mental interactions. Mistakes still occur, but
and has its correct shape. A major challenge is acids. It is not yet possible, however, to pre- improperly folded proteins can be recognized
to understand how proteins fold correctly—a dict accurately how a protein will fold from and degraded by enzymes called proteases.
puzzle known as the protein-folding problem. its amino acid sequence. The folding occurs The protein-folding problem has such impor-
Not only is this important from a purely sci- in a matter of seconds after the protein is tant implications for medicine, and is so
entific point of view, but a number of serious synthesized. The protein folds rapidly into challenging a scientific question, that a super-
neurodegenerative diseases result from protein its secondary structure and then more slowly computer with a huge memory is now being
misfolding. These include Alzheimer’s disease into its tertiary structure. These slower reac- used to help predict the three-dimensional
and the neurodegenerative diseases caused by tions are still poorly understood, but various structure of proteins from their amino acid
prions. If we could understand why proteins attractive and repulsive interactions between sequences. prions, pp. 12, 328

2.4 ■ Proteins Disaccharides

Proteins are made of amino acid subunits. Disaccharides are two monosaccharides joined together by covalent
bonds (figure 2.23). They include lactose, sucrose, and maltose (table 2.5).
Amino Acids
Polysaccharides
Amino acids have a central carbon atom, bonded to a carboxyl group,
an amino group, and a side chain. The side chain gives an amino acid Polysaccharides are large molecules composed of chains of monosac-
its characteristic properties (figure 2.13). Twenty major amino acids charide subunits or their derivatives. They include cellulose, starch,
function as subunits of proteins (figure 2.14). Most amino acids can exist glycogen, dextran, chitin, and agar (figure 2.24, table 2.5).
in two stereoisomers—d and l (figure 2.15). 2.6 ■ Nucleic Acids
Peptide Bonds Nucleic acids carry genetic information, which is decoded to produce
Peptide bonds join the amino group of one amino acid with the proteins.
carboxyl group of another (figure 2.16). DNA
Protein Structure (figure 2.17) The nucleotide sequence of DNA (deoxyribonucleic acid) encodes a
cell’s properties. The nucleotides have a nucleobase, deoxyribose, and
The primary structure of a protein is its amino acid sequence. The
a phosphate group (figures 2.25, 2.26, 2.27). DNA is a double-stranded
secondary structure is the three-dimensional shape of localized
helical molecule with a sugar phosphate backbone; the two strands
regions, characterized by helices and sheets. The tertiary structure
are antiparallel. The purine and pyrimidine nucleobases extend into
is the three-dimensional shape, usually either globular or fibrous. The
the center of the helix (figure 2.28a). The two strands of DNA are
quaternary structure is the shape that results from the interaction of
complementary—meaning there are G-C and A-T base pairs—and are
multiple polypeptide chains. Some proteins need chaperones to help
held together by hydrogen bonds between the nucleobases (figure 2.28b).
them fold into their functional shape.
RNA
Protein Domains RNA (ribonucleic acid) is involved in the process that decodes the
A protein domain is a region that folds into a stable structure indepen- information contained in DNA. RNA is a single-stranded molecule and
dently of other parts of the molecule (figure 2.18). its nucleotides contain uracil in place of thymine and ribose in place of
Substituted Proteins
deoxyribose (figures 2.20, 2.26).
Substituted proteins contain other molecules such as sugars and lipids, 2.7 ■ Lipids
bonded to the side chains of amino acids in the protein. Lipids are a diverse group of hydrophobic, non-polar molecules.
Protein Denaturation Simple Lipids
When bonds within a protein break, the protein changes shape and no Simple lipids contain carbon, hydrogen, and oxygen. Fats consist
longer functions; the protein is denatured (figure 2.19). of fatty acids linked to glycerol and may be liquid or solid at room
temperature (figure 2.29). Saturated fatty acids have no double bonds,
2.5 ■ Carbohydrates whereas unsaturated fatty acids have one or more double bonds.
Carbohydrates have carbon, hydrogen, and oxygen atoms in a ratio of Compound Lipids
approximately 1 : 2 : 1. Compound lipids contain fatty acids and glycerol as well as elements
Monosaccharides other than carbon, hydrogen, and oxygen. Phospholipids are essential
components of cytoplasmic membranes (figure 2.30).
Monosaccharides include the 5-carbon sugars ribose and deoxyribose
and the common 6-carbon sugars glucose, galactose, fruc-
(figure 2.20) Steroids
tose, and mannose (figure 2.21, table 2.5). Sugars can exist in two inter- Steroids are simple lipids that have a characteristic structure consisting
changeable forms: α and β (figure 2.22). of four connected rings (figure 2.31). The sterol cholesterol is an example.
38 Chapter 2 The Molecules of Life

Review Questions
Short Answer 8. Complementarity plays a major role in the structure of
1. Differentiate between an atom, a molecule, and a compound. a) proteins. b) lipids. c) polysaccharides.
2. Why is water a good solvent? d) DNA. e) RNA.
3. Which solution is more acidic, one with a pH of 4 or a pH of 5? 9. A bilayer is associated with
What is the concentration of H+ ions in each? The concentration of a) proteins. b) DNA. c) RNA.
OH- ions? d) complex polysaccharides. e) phospholipids.
4. Name the subunits of proteins, polysaccharides, and nucleic acids. 10. Isomers are associated with
5. Give an example of dehydration synthesis. Give an example of a 1. carbohydrates. 2. amino acids. 3. nucleotides.
hydrolysis reaction. How are these reactions related? 4. RNA. 5. fatty acids.
6. List four functions of proteins. a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5
7. What are the four levels of protein structure, and what is the distin-
guishing feature of each?
8. How do the two types of nucleic acids differ from one another in Applications
(a) composition, (b) size, and (c) function? 1. A group of prokaryotes known as thermophiles thrive at high
9. What are the two major groups of lipids? Give an example of each temperatures that would normally destroy other organisms. Yet
group. What feature is common to all lipids? these thermophiles cannot survive well at the lower tempera-
10. What features do all lipids share? tures normally found on the earth. Propose an explanation for
this observation.
Multiple Choice 2. Microorganisms use hydrogen bonds to attach to surfaces. Many
1. Choose the list that goes from the lightest to the heaviest: of the cells lose hold of the surface because of the weak nature
of these bonds. Contrast the benefits and disadvantages of using
a) proton, atom, molecule, compound, electron.
covalent bonds as a means of attaching to surfaces.
b) atom, proton, compound, molecule, electron.
c) electron, proton, atom, molecule, compound.
d) atom, electron, proton, molecule, compound. Critical Thinking +
e) proton, atom, electron, molecule, compound. 1. What properties of the carbon atom make it ideal as the key atom
2. The strongest chemical bonds between two atoms in solution are for all molecules in organisms?
a) covalent. b) ionic. 2. A biologist determined the amounts of several amino acids in two
c) hydrogen bonds. d) hydrophobic interactions. separate samples of pure protein. The data are shown here:
3. Dehydration synthesis is involved in the synthesis of all of the
following except
Amino Acid Leucine Alanine Histidine Cysteine Glycine
a) DNA. b) proteins. c) polysaccharides.
Protein A 7% 12% 4% 2% 5%
d) lipids. e) monosaccharides.
Protein B 7% 12% 4% 2% 5%
4. The primary structure of a protein relates to its
a) sequence of amino acids. b) length. c) shape.
The scientist concluded that protein A and protein B were the same
d) solubility. e) bonds between amino acids.
protein. Do you agree with this conclusion? Justify your answer.
5. Pure water has all of the following properties except
3. This table indicates the freezing and boiling points of several
a) polarity. b) ability to dissolve lipids. c) pH of 7.
molecules:
d) covalent joining of its atoms.
e) ability to form hydrogen bonds.
6. The macromolecules that are composed of carbon, hydrogen, and Molecule Freezing Point (°C) Boiling Point (°C)
oxygen in an approximate ratio of 1:2:1 are Water 0 100
a) proteins. b) lipids. c) polysaccharides. Carbon tetrachloride (CCl4) – 23 77
d) DNA. e) RNA. Methane (CH4) – 182 – 164
7. In proteins, α helices and β pleated structures are associated
with the
Carbon tetrachloride and methane are non-polar molecules. How
a) primary structure. b) secondary structure. does the polarity and non-polarity of these molecules explain why
c) tertiary structure. d) quaternary structure. the freezing and boiling points for methane and carbon tetrachlo-
e) multiprotein complexes. ride are so much lower than those for water?
3 Microscopy and Cell Structure
KEY TERMS
Capsule A distinct, thick gelatinous
material that surrounds some
microorganisms.
Lipopolysaccharide (LPS)
Molecule that makes up the outer
layer of the outer membrane of
Gram-negative bacteria.
Chemotaxis Movement of a
cell toward or away from a certain Peptidoglycan A macromolecule
chemical in the environment. that provides strength to the cell
wall; it is found only in bacteria.
Cytoplasmic Membrane A
phospholipid bilayer embedded with Periplasm The gel-like material
proteins that surrounds the cytoplasm that fills the region between the
and defines the boundary of the cell. cytoplasmic membrane and the
outer membrane of Gram-negative
Endospore An extraordinarily bacteria.
resistant dormant cell produced by
some bacteria. Pili Cell surface structures that
allow cells to adhere to certain
Flagellum A type of structure used surfaces; some types are involved in
for cell movement. a mechanism of DNA transfer.
Gram-Negative Bacteria Bacteria Plasmid Extrachromosomal
that have a cell wall characterized DNA molecule that replicates
by a thin layer of peptidoglycan independently of the chromosome.
surrounded by an outer membrane;
Ribosome Structure involved in
when Gram stained, these cells are
protein synthesis.
Bacterial cells (color-enhanced scanning electron micrograph). pink.
Transport Systems Mechanisms
Gram-Positive Bacteria Bacteria used to transport nutrients and
that have a cell wall characterized by other small molecules across the
A Glimpse of History a thick layer of peptidoglycan; when cytoplasmic membrane.
Gram stained, these cells are purple.
Hans Christian Joachim Gram (1853–1938) was a Danish physician
working in a laboratory at the morgue of the City Hospital in Berlin,
microscopically examining the lungs of patients who had died of pneumo-
nia. He was working under the direction of Dr. Carl Friedlander, who was
trying to identify the cause of pneumonia by studying patients who had
The similarities and differences between these two basic cell types
died of it. Gram’s task was to stain the infected lung tissue to make the
bacteria easier to see under the microscope. Strangely, one of the meth-
are important from a scientific standpoint. They also have signifi-
ods he developed did not stain all bacteria equally; some types retained cant consequences to human health. For example, chemicals that
the first dye applied in this multistep procedure, whereas others did not. interfere with processes unique to prokaryotic cells can be used to
Gram’s staining method revealed that two different kinds of bacteria were selectively destroy bacteria without harming humans. prokaryotic
causing pneumonia, and these types retained the dye differently. We now cells, p. 9 eukaryotic cells, p. 10
recognize that this important staining method, called the Gram stain, effi- Prokaryotic cells are generally much smaller than most
ciently identifies two large, distinct groups of bacteria: Gram-positive and eukaryotic cells—a trait that carries with it certain advantages
Gram-negative. The staining outcome reflects a fundamental difference in as well as disadvantages. On one hand, their high surface area
the structure and chemistry of the cell walls of these two groups, and is a relative to low volume makes it easier for these cells to take in
key test in the initial identification of bacterial species. nutrients and excrete waste products. Because of this, they can
multiply much faster than eukaryotic cells. On the other hand,

I
magine the astonishment Antony van Leeuwenhoek must have their small size makes them vulnerable to a variety of threats,
felt in the 1600s when he first observed microorganisms with including predators, parasites, and competitors. To cope, pro-
his handcrafted microscopes, instruments that could magnify karyotes have evolved many unique features that increase their
images approximately 300-fold. Even today, observing diverse chances of survival.
microbes interacting in a sample of pond water can provide enor- Eukaryotic cells are much more complex than prokaryotic cells.
mous education and entertainment. Not only are they larger, but many of their cellular processes take
Microscopic study of cells has revealed two fundamental place within membrane-bound compartments. Eukaryotic cells are
types: prokaryotic and eukaryotic. The cells of all members of the defined by the presence of one of these—the nucleus. Although
domains Bacteria and Archaea are prokaryotic. In contrast, cells eukaryotic cells have many of the same characteristics as prokaryotic
of all animals, plants, protozoa, fungi, and algae are eukaryotic. cells, some structures and processes are fundamentally different.
39
40 Chapter 3 Microscopy and Cell Structure

MICROSCOPY AND CELL MORPHOLOGY

3.1 ■ Microscopic Techniques: 1931, can magnify images in excess of 100,000×, revealing many
fine details of cell structure. A major advancement came in the
The Instruments 1980s with the development of the atomic force microscope,
which allows scientists to produce images of individual atoms on
Learning Outcomes
a surface.
1. Describe the importance and principles of magnification,
resolution, and contrast in microscopy.
2. Compare and contrast light microscopes, electron microscopes,
and atomic force microscopes.
Principles of Light Microscopy:
The Bright-Field Microscope
One of the most important tools for studying microorganisms is In light microscopy, light passes through a specimen and then
the light microscope, which uses visible light for observing through a series of magnifying lenses. The most common type of
objects. These instruments can magnify images approximately light microscope is the bright-field microscope, which evenly
1,000× (1,000-fold), making it relatively easy to observe cell size, illuminates the field of view. Characteristics of this and other
shape, and motility. The electron microscope, introduced in microscopes are summarized in table 3.1.

TABLE 3.1 A Summary of Microscopic Instruments and Their Characteristics

Instrument Mechanism Comment

Light Microscopes Visible light passes through a series of lenses to Relatively easy to use; considerably less expensive
produce a magnified image. than electron and atomic force microscopes.

Bright-field Illuminates the field of view evenly. Most common type of microscope.

Dark-field Light is directed toward the specimen at an angle. Makes unstained cells easier to see; organisms
stand out as bright objects against a dark
background.

Phase-contrast Increases contrast by amplifying differences in Makes unstained cells easier to see.
refractive index.
 Part I Life and Death of Microorganisms 41

Instrument Mechanism Comment

Differential interference contrast Two light beams pass through the specimen and The image of the specimen appears three-
then recombine. dimensional.

Fluorescence Projects ultraviolet light, causing fluorescent Used to observe cells stained or tagged with a
molecules in the specimen to emit longer fluorescent dye.
wavelength light.

Scanning laser Mirrors scan a laser beam across successive regions Used to construct a three-dimensional image of
and planes of a specimen. From that information, a a structure; provides detailed sectional views of
computer constructs an image. intact cells.

Electron Microscopes Electron beams are used in place of visible light to Can clearly magnify images 100,000×.
produce the magnified image.
Transmission Transmits a beam of electrons through a specimen. Elaborate specimen preparation is required.

Scanning A beam of electrons scans back and forth over the Used for observing surface details; produces a
surface of a specimen. three-dimensional effect.

Atomic Force Microscope A probe moves in response to even the slightest Produces a map showing the bumps and valleys of
force between it and the sample. the atoms on the surface of the sample.
42 Chapter 3 Microscopy and Cell Structure

Magnification
The modern light microscope, called a compound microscope,
has two types of magnifying lenses: an objective lens and an
ocular lens (figure  3.1). In combination, these lenses visually
enlarge an object by a factor equal to the product of each lens’
magnification. For example, a structure is magnified 1,000-fold
when viewed through a 10× ocular lens in conjunction with a
100× objective lens. Most compound microscopes have a selec-
tion of objective lenses that are of different powers—typically 4×,
10×, 40×, and 100×. This makes a choice of different magnifica-
tions possible with the same instrument. Light microscope (450×) Electron microscope (450×)
The condenser lens, positioned between the light source and FIGURE 3.2 Resolving Power These images of an onion root tip
the specimen, does not affect the magnification. It focuses the magnified 450× illustrate the difference in resolving power between
light on the specimen. a light microscope and an electron microscope. Note the difference
in the degree of detail that can be seen at the same magnification.
Resolution ? Which type of microscope—a light microscope or an electron
The usefulness of a microscope depends primarily on its resolving microscope—has the higher resolving power?
power, which determines how much detail can actually be seen
(figure 3.2). Resolving power is a measure of the ability to distin- a medium of one refractive index (a measure of the relative speed
guish two objects that are very close together. It is defined as the of light as it passes through the medium) to another. If refraction
minimum distance between two points at which those points can occurs, some light rays will miss the relatively small openings of
still be observed as separate objects. higher-power objective lenses. The oil prevents refraction because
The resolving power of a microscope depends on the quality it has nearly the same refractive index as glass.
and type of lens, the wavelength of the light, the magnification,
and how the specimen has been prepared. The maximum resolving Contrast
power of the best light microscope is 0.2 μm. This is sufficient to
The amount of contrast affects how easily cells can be seen. As
see the general morphology of a prokaryotic cell but too low to
an example, bacteria are essentially transparent against a bright
distinguish an object the size of most viruses.
colorless background, so the lack of contrast makes them harder
To obtain maximum resolution when using certain high-power
objectives such as the 100× lens, immersion oil must be used to
displace the air between the lens and the specimen. This prevents
the refraction (bending of light rays) that occurs when light passes
from glass to air (figure 3.3). Light rays bend when they pass from
Objective lens

Ocular lens
(eyepiece)
Magnifies the Objective lens
image, usually A selection of lens
10-fold (10×). options provide
different
magnifications. The Air Oil
Specimen total magnification is
stage the product of the
magnifying power of
Condenser the ocular lens and
the objective lens. Slide Light source
Focuses
the light. (a) (b)
Light source
Iris diaphragm FIGURE 3.3 Refraction As light passes from one medium to
Controls the Rheostat another, the light rays may bend, depending on the refractive index of
amount of light Controls the the two media. (a) The pencil in water appears bent because the
that enters the brightness of the
refractive index of water is different from that of air. (b) Light rays
objective lens. light.
bend as they pass from air to glass because of the different refractive
FIGURE 3.1 A Modern Light Microscope The compound indexes of these media. Immersion oil and glass have the same
microscope uses a series of magnifying lenses. refractive index, and therefore the light rays are not bent.
? What are the two sets of magnifying lenses called, and how do ? What would the pencil in part (a) look like if oil were in the glass
these relate to total magnification? instead of water?
 Part I Life and Death of Microorganisms 43

to see (see figure 11.17). One way to overcome this difficulty is to

stain the cells with one of the various dyes that will be discussed
shortly. The staining process kills microbes, however, so living
cells cannot be observed.

Light Microscopes
That Increase Contrast
Special light microscopes that increase the contrast between
microorganisms and their surroundings overcome some of the
difficulties of observing unstained cells. They are invaluable
when examining characteristics of living organisms such as
motility. When observing live organisms, the specimen is pre-
pared as a wet mount—a drop of liquid on which a coverslip has
been placed.

The Dark-Field Microscope 25 µm

Cells viewed through a dark-field microscope stand out as bright FIGURE 3.5 Phase-Contrast Photomicrograph Paramecium
objects against a dark background (figure  3.4). The microscope bursaria containing endosymbiotic Chlorella (a green alga).
works in the same way that a beam of light shining into a dark ? How does a phase-contrast microscope increase contrast?
room makes dust visible. For dark-field microscopy, a special
mechanism directs light toward the specimen at an angle, so that
only light scattered by the specimen enters the objective lens. slight difference between the refractive index of dense material
A simple “dark-field stop” can be attached to the condenser lens and that of the surrounding medium. As light passes through mat-
of a bright-field microscope to temporarily convert it to a dark- erial, it is refracted slightly differently than when it passes through
field microscope. its surroundings. Special optical devices increase those differ-
ences, thereby enhancing the contrast.
MicroByte
Dark-field microscopy is used to see Treponema pallidum, which The Differential Interference
causes syphilis; bright-field microscopy will not work because the Contrast Microscope (DIC)
cells are thin and stain poorly. The differential interference contrast microscope (DIC) causes
the image of the specimen to appear three-dimensional (figure 3.6).
The Phase-Contrast Microscope This microscope, like the phase-contrast microscope, depends on
The phase-contrast microscope makes cells and other dense
material appear darker (figure 3.5). It does this by amplifying the

Filamentous alga
(Spirogyra sp.)

Colonial alga
(Volvox sp.)

25 µm 30 µm

FIGURE 3.4 Dark-Field Photomicrograph Volvox (sphere) and FIGURE 3.6 Differential Interference Contrast (DIC) Photomicro-
Spirogyra (filaments), both of which are eukaryotes. graph Paramecium multimicronuclatum, a protozoan.
? How does a dark-field microscope increase contrast? ? How does a DIC microscope increase contrast?
44 Chapter 3 Microscopy and Cell Structure

differences in refractive index as light passes through different

materials. It has a device for separating light into two beams that
pass through the specimen and then recombine. The light waves
are out of phase when they recombine, thereby creating the three-
dimensional appearance of the image.

Other Light Microscopes

Various other light microscopes have special characteristics that
make them useful in certain situations.
Fluorescence Microscopes
Fluorescence microscopes are used to observe cells or other
materials that are either naturally fluorescent or stained or tagged
with fluorescent dyes (figure 3.7). Fluorescent molecules absorb
light at one wavelength (usually ultraviolet light) and then emit (a) (b)
light of a longer wavelength. The types and characteristics of fluo- FIGURE 3.8 Scanning Laser Microscopy (a) Confocal photo-
rescent dyes and tags will be discussed shortly. fluorescent dyes micrography of fava bean mitosis. (b) Regular photomicrograph.
and tags, p. 49
? How is multiphoton microscopy different from confocal microscopy?
Most fluorescence microscopes used today are epifluorescence
microscopes, meaning they project ultraviolet light onto the speci-
men rather than through it. They then capture the light emitted by
the fluorescent molecules to form the image, allowing fluorescent then assembles the data and constructs a three-dimensional image,
cells to stand out as bright objects against a dark background. which is displayed on a screen. In effect, this microscope is a minia-
Because the light does not need to travel through the specimen, cells ture computerized axial tomography (CAT) scan for cells.
attached to soil or other opaque particles can be observed. Multiphoton microscopy is similar to confocal microscopy,
but lower energy light is used. This light is less damaging to cells,
Scanning Laser Microscopes so time-lapse images of live cells can be obtained. In addition, the
A scanning laser microscope (SLM) can be used to get detailed light penetrates deeper, making it possible to get interior views of
interior views of intact cells (figure 3.8). Frequently, the specimens relatively thick structures.
are first stained or tagged with a fluorescent dye. By using fluores-
cent tags that bind only to certain compounds, the precise cellular Electron Microscopes
location of those compounds can be determined. Some scanning Electron microscopy is in some ways comparable to light micros-
laser microscopes can also be used to make three-dimensional copy. Rather than using glass lenses, visible light, and the eye to
images of microbial communities or other thick structures. observe the specimen, an electron microscope uses electromag-
In confocal microscopy, lenses focus a laser beam to illuminate netic lenses, electrons, and a fluorescent screen to produce the
a given point on one vertical plane of a specimen. Mirrors then scan magnified image (figure  3.9). That image can be captured on
the laser beam across the specimen, illuminating successive regions photographic film to create an electron micrograph. Sometimes
and planes until the entire specimen has been scanned. Each plane the black-and-white images are artificially enhanced with color to
corresponds to an image of one fine slice of the specimen. A computer add visual clarity. electrons, p. 17
Electrons have a wavelength about 1,000 times shorter than
visible light, so the resolving power of electron microscopes is
about 1,000-fold more than light microscopes—about 0.3 nano-
meters (nm) or 0.3 ×10–3 μm (see figure  1.13). Consequently,
considerably more detail can be observed. These instruments can
clearly magnify an image 100,000×. One of the biggest draw-
backs of the microscope is that the lenses and specimen must all
be in a vacuum. Otherwise, the molecules composing air would
interfere with the path of the electrons. The need for a vacuum
results in an expensive, bulky unit, and requires substantial and
complex specimen preparation. It also means that electron micros-
copy cannot be used to observe the activities of living cells.
Transmission Electron Microscopes
A transmission electron microscope (TEM) is used to observe fine
10 µm
details of cell structure (figure 3.10). It works by directing a beam of
FIGURE 3.7 Fluorescence Photomicrograph A rod-shaped electrons that will either pass through the specimen or be scattered,
bacterium tagged with fluorescent marker. depending on the density of the region. The darker areas of the
? What is an epifluorescence microscope? resulting image correspond to the denser portions of the specimen.
 Part I Life and Death of Microorganisms 45

Light Microscope Transmission Electron

Microscope

Electron
Lamp gun

Glass Condenser Electromagnet

lens

Light Electron beams

rays
(a) 1 µm

Specimen

Glass Objective Electromagnet

lens

Image

Glass Ocular Electromagnet

lens (b) 1 µm

FIGURE 3.10 Transmission Electron Photomicrograph A rod-

shaped bacterium prepared by (a) thin-sectioning; (b) freeze-etching.
? How is thin-sectioning different from freeze-etching?

Eye Viewing screen

temperatures, thereby avoiding some of the sample preparation
FIGURE 3.9 Comparison of the Principles of Light and processes that damage cells. A variation called cryo-electron
Electron Microscopy For the sake of comparison, the light source
for the light microscope has been inverted (the light is shown at the
tomography compiles images taken at different angles to create
top and the ocular lens at the bottom). three-dimensional images of specimens.
? Some electron micrographs are “color enhanced.” What does this Scanning Electron Microscopes
mean?
A scanning electron microscope (SEM) is used to observe sur-
face details of cells (figure 3.11). A beam of electrons scans back
A process called thin-sectioning is used to view details of a and forth over the surface of a specimen coated with a thin film of
specimen’s internal structures. First, the sample is treated with a metal. As those beams move, electrons are released from the
preservative and dehydrated before embedding it in plastic. Once specimen and reflected back into the viewing chamber. This
embedded, the sample can be cut into exceptionally thin slices reflected radiation is observed with the microscope. Relatively
with a diamond or glass knife and then stained with heavy metals. large specimens can be viewed, and a dramatic three-dimensional
Even a single bacterial cell must be cut into slices to be viewed via effect is observed with the SEM.
TEM. Unfortunately, the procedure can severely distort cells,
causing artifacts (artificial structures introduced as a result of the
process). Distinguishing actual cell components from artifacts that
result from specimen preparation is a major concern.
A method called freeze-fracturing is used to observe the shape
of structures within a cell. The specimen is rapidly frozen and then
fractured by hitting it with a knife blade. The cells break open, usu-
ally along the middle of internal membranes. Next, the surface of
the section is coated with a thin layer of carbon to create a replica
of the surface. This replica is then examined in the electron micro-
scope. A variation of freeze-fracturing is freeze-etching. In this
process, the frozen surface exposed by fracturing is dried slightly 2 µm

under vacuum, which allows underlying regions to be exposed. FIGURE 3.11 Scanning Electron Photomicrograph A rod-shaped
A newer method of preparing and observing specimens with bacterium.
an electron microscope is cryo-electron microscopy (cryo-EM). ? In what way is scanning electron microscopy different from
This method involves rapidly freezing the specimen at very low transmission electron microscopy?
46 Chapter 3 Microscopy and Cell Structure

3.2 ■ Microscopic Techniques:

Dyes and Staining
Learning Outcomes
3. Describe the principles of the Gram stain and the acid-fast stain.
4. Describe the techniques used to observe capsules, endospores, and
flagella.
5. Describe the benefits of using fluorescent dyes and tags.

Observing living microorganisms with the bright-field microscope

can be difficult because the cells often move around quickly and
0.3 µm are nearly transparent. This problem can be avoided by immobiliz-
FIGURE 3.12 Atomic Force Photomicrograph A DNA fragment. ing the cells and staining them with dyes. To do this, a drop of
The bright peaks are enzymes attached to the DNA. liquid containing the organism is first placed on a microscope
? How does the resolving power of atomic force microscopy slide and allowed to dry (figure  3.13). The resulting specimen
compare to that of electron microscopy? forms a film, or smear. Next, the slide is passed over a flame to fix
(attach) the cells to the slide. Dye is then applied and the excess
washed off with water. If only a single dye is used, the procedure
Atomic Force Microscopes is called simple staining (table 3.2).
An atomic force microscope (AFM) produces detailed images of Staining procedures typically use basic dyes, meaning the
surfaces (figure 3.12). The resolving power is much greater than dyes carry a positive charge. These dyes stain cells because they
that of an electron microscope, and the samples do not need the are attracted to the many negatively charged cellular components.
special preparation required for electron microscopy. In fact, the Examples of basic dyes include methylene blue, crystal violet,
instrument can inspect samples either in air or submerged in liquid. safranin, and malachite green.
The mechanics of AFM can be compared to that of a stylus Although acidic dyes do not stain cells, they can be used for
mounted on the arm of a record player. A very sharp probe (stylus) negative staining, a procedure that colors the background. The
moves across the sample’s surface, “feeling” the bumps and valleys of cells repel the negatively charged dye, allowing the colorless cell
the atoms. As the probe scans the sample, a laser measures its motion, to stand out against the background. An advantage of negative
and a computer produces a surface map of the sample. atom, p. 17 staining is that it can be done as a wet mount. This avoids the heat-
fixing step, a process that can distort the shape of the cells.
MicroAssessment 3.1
The usefulness of a microscope depends on its resolving power. The most Differential Staining
common type of microscope is the bright-field microscope. Dark-field, Differential staining is used to distinguish different groups of
phase-contrast, and DIC microscopes increase the contrast between a
microorganism and its surroundings. The fluorescence microscope is used
bacteria. The two most frequently used differential staining tech-
to observe microbes stained with special dyes. Scanning laser microscopes niques are the Gram stain and the acid-fast stain.
are used to construct three-dimensional images of thick structures. Electron
microscopes can magnify images 100,000×. The atomic force microscope
produces detailed images of surfaces.
1. Why must oil be used to obtain the best resolution with a 100× lens?
2. What are some drawbacks of electron microscopes?
3. If an object being viewed under the phase-contrast microscope
has the same refractive index as the background material, how
would it appear? +

Spread thin film of Allow to air dry. Pass slide Flood the smear with Examine with microscope.
specimen over slide. through stain, rinse, and dry.
flame to
heat-fix
specimen.
FIGURE 3.13 Staining Bacteria
? What is the purpose of heating the smear?
 Part I Life and Death of Microorganisms 47

TABLE 3.2 A Summary of Stains and Their Characteristics

Stain Characteristic
Simple Stains A basic dye is used to stain cells. Easy way to increase the contrast between otherwise colorless cells and a
transparent background.
Differential Stains A multistep procedure is used to stain cells and distinguish one group of microorganisms from another.
Gram stain Used to separate bacteria into two major groups, Gram-positive and Gram-negative. The staining characteristics
of these groups reflect a fundamental difference in the chemical structure of their cell walls. This is by far the
most widely used staining procedure.
Acid-fast stain Used to detect organisms that do not easily take up stains, such as members of the genus Mycobacterium.
Special Stains Stain specific structures inside or outside of cells.
Capsule stain These are often negative stains that take advantage of the fact that viscous capsules do not take up certain
stains; the capsules stand out against a stained background.
Endospore stain Stains endospores, a type of dormant cell that does not readily take up stains. Endospores are produced by
Bacillus and Clostridium species.
Flagella stain The staining agent adheres to and coats the otherwise thin flagella, making them visible with the light microscope.
Fluorescent Dyes and Tags Fluorescent dyes and tags absorb ultraviolet light and then emit light of a longer wavelength. They are used in
conjunction with a fluorescence microscope.
Fluorescent dyes Some fluorescent dyes bind to compounds found in all cells; others bind to compounds specific to only certain types
of cells.
Fluorescent tags Antibodies to which a fluorescent molecule has been attached are used to tag specific molecules.

Gram Stain Gram staining involves four basic steps (figure 3.14).

The Gram stain is by far the most widely used procedure for stain- 1. The smear is first flooded with the primary stain, crystal
ing bacteria. The basis for it was developed over a century ago by violet in this case. The primary stain is the first dye applied in
Dr. Hans Christian Gram (see A Glimpse of History). He showed differential staining and generally stains all cells.
that bacteria can be separated into two major groups: Gram- 2. The smear is rinsed to remove excess dye and then flooded
positive bacteria and Gram-negative bacteria. We now know with a solution called Gram’s iodine. The iodine combines
that the difference in the staining properties of these two groups with the crystal violet to form a dye-iodine complex, thereby
reflects a fundamental difference in the structure of their cell walls. decreasing the solubility of the dye within the cell.

Steps in Staining State of Bacteria Appearance

1 Crystal violet Cells stain purple.

(primary stain)

2 Iodine Cells remain purple.

(mordant)

3 Alcohol Gram-positive cells

(decolorizer) remain purple;
Gram-negative cells
become colorless.

4 Safranin Gram-positive cells

(counterstain) remain purple;
Gram-negative cells
appear pink.

(a) (b) 10 µm

FIGURE 3.14 Gram Stain (a) Steps in the Gram stain procedure. (b) Results of a Gram stain. The Gram-positive cells (purple) are
Staphylococcus aureus; the Gram-negative cells (reddish-pink) are Escherichia coli.
? Which step of the Gram stain is most critical with respect to timing?
48 Chapter 3 Microscopy and Cell Structure

3. The stained smear is rinsed, and then a decolorizing

agent—95% alcohol or a mixture of alcohol and acetone—is
briefly added. These chemicals quickly remove the dye-iodine
complex from Gram-negative, but not Gram-positive, bacteria.
4. A counterstain is then applied to give a different color to the
now colorless Gram-negative bacteria. For this purpose, the
red dye safranin is used. This dye enters Gram-positive cells
as well, but because they are already purple, it makes little
difference to their color.
To obtain reliable results, the Gram stain must be done prop-
erly. One common mistake is to decolorize a smear for the incor-
rect time period. Even Gram-positive cells can lose the crystal
violet-iodine complex during prolonged decolorization. When
this happens, over-decolorized Gram-positive cells will appear
pink after counterstaining. In contrast, under-decolorizing results FIGURE 3.15 Acid-Fast Stain Mycobacterium species retain the
in Gram-negative cells appearing purple. Another important red primary stain, carbolfuchsin. Counterstaining with methylene blue
consideration is the age of the culture. As bacterial cells age, colors the non-acid-fast cells blue.
they lose their ability to retain the crystal violet–iodine dye com- ? What characteristic of Mycobacterium cells makes them acid-fast?
plex. As a result, cells from old cultures may appear pink. Thus,
Gram staining fresh cultures (less than 24 hours old) gives more
reliable results. Capsule Stain
A capsule is a gel-like layer that surrounds certain microbes. This
Acid-Fast Stain layer has a protective function and often increases an organism’s
The acid-fast stain is a procedure used to detect a small group of pathogenicity. Capsules stain poorly, so capsule stains often use
organisms that do not readily take up dyes. Among these are mem- negative staining to make them visible. In one common method,
bers of the genus Mycobacterium, including a species that causes India ink is added to a suspension of cells to make a wet mount.
tuberculosis and one that causes Hansen’s disease (leprosy). The The fine particles of ink darken the background, allowing the cap-
cell wall of these bacteria contains high concentrations of mycolic sule to stand out as a clear area surrounding a cell (figure 3.16).
acid, a waxy fatty acid that prevents uptake of dyes such as those capsule, p. 62 pathogen, p. 6

used in Gram staining. Therefore, harsh methods are needed to

stain these organisms. Once stained, however, these same cells are Endospore Stain
very resistant to decolorization. Because mycobacteria are among Members of certain genera, including Bacillus and Clostridium,
the few organisms that retain the dye in this procedure, acid-fast form a special type of resistant, dormant cell called an endospore.
staining can be used to presumptively identify them in clinical These structures do not stain with the Gram stain, but they can
specimens. tuberculosis, p. 502 Hansen’s disease, p. 650 fatty acid, often be seen as clear, smooth objects within stained cells.
p. 33 endospore, p. 67
Acid-fast staining, like Gram staining, requires multiple
steps. The primary stain is carbolfuchsin, a red dye. In the classic
procedure, the dye-flooded smear is heated, which facilitates
staining. A  current variation does not include heat, but instead
uses a concentrated solution of dye for a longer period of time.
The slide is then rinsed briefly to remove excess dye before being
flooded with acid-alcohol, a strong decolorizing agent. This agent
removes the carbolfuchsin from tissue cells and most bacteria;
those few species that retain the dye are called acid-fast.
Methylene blue is then used as a counterstain. As a result of the
staining procedure, acid-fast organisms are bright reddish-pink,
making them easy to distinguish from the blue non-acid-fast cells
(figure 3.15).

10 µm
Special Stains to Observe Cell Structures FIGURE 3.16 Capsule Stain Cryptococcus neoformans, an
Special procedures can also be used to stain specific structures encapsulated yeast. The capsules stand out against the India
inside or outside the cell. The function of these structures will be ink–stained background.
discussed in more depth later in the chapter. ? How is the India ink capsule stain an example of a negative stain?
 Part I Life and Death of Microorganisms 49

10 µm 1 µm

FIGURE 3.17 Endospore Stain Endospores retain the primary FIGURE 3.18 Flagella Stain The staining agent adheres to and
stain, malachite green. Counterstaining with safranin colors other cells coats the flagella. This increases their diameter so they can be seen
red. with the light microscope.
? What color would Escherichia coli cells be with the endospore ? How can the flagella stain be helpful in identifying bacteria?
stain shown in the photo?

To make endospores easier to see, an endospore stain is thin to be seen with the light microscope. The flagella stain uses
used. This multistep procedure often uses malachite green as a a substance that allows the staining agent to adhere to and coat the
primary stain, with gentle heating to facilitate uptake of the dye by thin flagella, making them visible using light microscopy. Not all
endospores. The smear is then rinsed with water, which removes prokaryotic cells have flagella, but those that do can have them in
the dye from everything but the endospores. The smear is then different arrangements around a cell, so the presence and distribu-
counterstained, most often with the red dye safranin. Using this tion of these appendages can be used as identifying features
method, the endospores will be green while all other cells will be (figure 3.18). flagella, p. 63
pink (figure 3.17).

Fluorescent Dyes and Tags

Flagella Stain Fluorescence can be used to observe total cells, a subset of cells,
Flagella are appendages that provide the most common mecha- or cells with certain proteins on their surface, depending on the
nism of motility for prokaryotic cells, but they are ordinarily too procedure employed (figure 3.19).

(a) 10 µm (b) 10 µm (c) 10 µm

FIGURE 3.19 Fluorescent Dyes and Tags (a) Dyes that cause live cells to fluoresce green and dead ones red. (b) Auramine is used to stain
Mycobacterium species in a modification of the acid-fast technique. (c) Fluorescent antibodies tag specific molecules—in this case, the antibody
binds to a molecule unique to Streptococcus pyogenes.
? How can fluorescent dyes and tags be used to identify bacteria?
50 Chapter 3 Microscopy and Cell Structure

Fluorescent Dyes
Coccus Rod (bacillus)
Some fluorescent dyes bind to compounds found in all cells
whereas others bind only to certain components. As an example,
acridine orange binds DNA, making it useful for determining the
total number of microbes in a sample. In contrast, calcofluor white
binds to a cell wall component of fungi and certain bacteria, caus-
ing those organisms to fluoresce bright blue. The fluorescent dyes
auramine and rhodamine bind to the mycolic acid in the cell walls
of Mycobacterium species, making the dyes useful in a staining
procedure similar to the acid-fast stain. Other fluorescent dyes are
changed by cellular processes, allowing microbiologists to distin-
guish between living and dead cells. mycolic acid, p. 503

Immunofluorescence (a) 1 µm (b) 11.4 µm

Immunofluorescence is a technique used to tag specific proteins
with a fluorescent compound. By tagging a protein unique to a Vibrio Spirillum
given microbe, immunofluorescence can be used to detect that
organism. Immunofluorescence uses an antibody to deliver the
fluorescent tag (see figure  18.8). Antibodies, and how they are
made, will be described in chapter 15. antibody, pp. 355, 359

MicroAssessment 3.2
Dyes are used to stain cells so they can be seen against an unstained
background. The Gram stain is the most commonly used differential
stain. The acid-fast stain is used to detect Mycobacterium species.
Specific dyes and techniques are used to observe cell structures such
as capsules, endospores, and flagella. Fluorescent dyes and tags can
be used to observe total cells, a subset of cells, or cells that have
certain proteins on their surface. (c) 15 µm (d) 15 µm
4. What are the functions of a primary stain and a counterstain?
5. Describe one error in the staining procedure that would result in Spirochete
a Gram-positive bacterium appearing pink.
6. What color would a Gram-negative bacterium be in an acid-fast
stain? +

3.3 ■ Morphology FIGURE 3.20 Shapes of

of Prokaryotic Cells Common Bacteria (a) Coccus;
(b) rod; (c) vibrio; (d) spirillum;
(e) spirochete. Electron micro-
Learning Outcomes graphs.
6. Describe the common bacterial shapes and groupings, and their
? What are the two most
significance. common shapes of bacteria?
7. Describe two multicellular associations of bacteria. (e) 7.5 µm

Prokaryotic cells come in a variety of simple shapes and often

form characteristic groupings. Some aggregate, living as multicel- is often called a coccobacillus. The descriptive term “bacillus”
lular associations. should not be confused with Bacillus, the name of a genus.
Although members of the genus Bacillus are rod-shaped, so are
many other bacteria, including Escherichia coli.
Shapes Cells come in a variety of other shapes. A short, curved rod
Most common bacteria are one of two shapes: spherical, called a is a vibrio (plural: vibrios), whereas a curved rod long enough
coccus (plural: cocci), or cylindrical, called a rod (figure 3.20). A to form spirals is a spirillum (plural: spirilla). A long, helical
rod-shaped bacterium is sometimes called a bacillus (plural: cell with a flexible cell wall and a unique mechanism of motility
bacilli), and one so short that it can easily be mistaken for a coccus is a spirochete. Bacteria that characteristically vary in their
 Part I Life and Death of Microorganisms 51

shape are pleomorphic (pleo meaning “many” and morphic refer- In their natural habitat, most bacteria on surfaces live in com-
ring to shape). munities called biofilms. Details about these communities are
The greatest diversity in shapes is found in low-nutrient described in chapter 4. biofilms, p. 84
aquatic environments. Cells there often have a large surface area,
which helps them absorb nutrients more easily (figure 3.21). For MicroAssessment 3.3
example, some aquatic bacteria have cytoplasmic extensions, giv-
ing them a starlike appearance. Square, tilelike archaeal cells have Most common prokaryotes are cocci or rods, but other shapes include
vibrios, spirilla, and spirochetes. Cells may form characteristic
been found in salt ponds.
groupings such as chains or clusters. Some form multicellular
associations.
Groupings 7. What shape are Escherichia coli cells?
Most prokaryotes divide by binary fission, a process in which one 8. What determines whether a group of dividing cells will form
chains or clusters? +
cell divides into two. Cells often stick to each other following
division, forming characteristic groupings (figure 3.22).
Cells that divide in one plane can form chains of varying
length. Cocci that typically occur in pairs are routinely called
diplococci. An important clue in the identification of Neisseria
gonorrhoeae is its characteristic diplococcus arrangement. Other Chains
cells form long chains, a characteristic typical of some members
of the genus Streptococcus.
Cocci often divide in more than one plane. Those that divide
in perpendicular planes form cubical packets. Members of the
genus Sarcina form such packets. Cocci that divide in several
planes at random may form clusters. Staphylococcus species, Diplococcus
which typically form grapelike clusters, are an example.
Cell divides
in one plane.
Multicellular Associations
Some prokaryotes characteristically live as multicellular associa-
Chain of cocci
tions. For example, members of a group of bacteria called myxo-
bacteria glide over moist surfaces, forming swarms of cells that (a)
move as a pack. The cells release enzymes, and, as a pack, they
degrade organic material, including other bacterial cells. When Packets
water or nutrients become limiting, the cells come together to
form a structure called a fruiting body, which is visible to the
naked eye (see figure 11.16). myxobacteria, p. 268
Cell divides
in two or more planes
perpendicular to one Packet
another.

(b)

Clusters

Cell divides
in several planes at
random.
Cluster

(a) 1 µm (b) 1 µm (c)

FIGURE 3.21 Diverse Shapes of Aquatic Prokaryotes FIGURE 3.22 Typical Cell Grouping (a) Chains; (b) packets;
(a) Square, tilelike archaeal cell. (b) Ancalomicrobium adetum, (c) clusters.
a bacterium that has a starlike appearance.
? What aspect of bacterial cell division determines the characteristic
? Why would aquatic microbes need maximal surface area? cell arrangements?
 Focus Figure

Pilus

Ribosomes
Cytoplasm

Chromosome
(DNA)
Nucleoid

Cell wall
Flagellum

(b) 0.5 µm

Capsule Cell wall Cytoplasmic

membrane
(a)

FIGURE 3.23 Typical Structures of a Prokaryotic Cell (a) Diagrammatic representation. Note that archaea rarely have capsules. (b) Electron
micrograph.
? How does the function of the cytoplasmic membrane differ from that of the cell wall?

THE PROKARYOTIC CELL

Knowing the structure and function of the various prokaryotic MicroByte
cell components is essential for understanding how microbes Our bodies defenses have “alarm systems” that recognize compounds
survive. The information is particularly relevant from a medical unique to bacteria, using them as indicators of invading microbes.
standpoint because characteristics unique to bacteria are poten-
tial targets for antibacterial medications. By interfering with
these targets, we can selectively kill bacteria or inhibit their 3.4 ■ The Cytoplasmic Membrane
growth without harming the human host. In addition, features
found in only select groups of prokaryotes can be used to iden- Learning Outcomes
tify different species. 8. Describe the structure and chemistry of the cytoplasmic membrane,
The structures of a prokaryotic cell allow it to survive and focusing on how it relates to membrane permeability.
multiply in a given environment. Some of the components are 9. Describe how the cytoplasmic membrane is involved with proton
essential for growth and therefore found in all prokaryotic cells, motive force.
but others are optional. While these optional parts are not required
in the protected confines of a laboratory, cells lacking them might The cytoplasmic membrane (or plasma membrane) is a thin,
not survive the competitive environment of the outside world. delicate structure that surrounds the cytoplasm and defines the
The surface layers of a prokaryotic cell, called the cell envelope, boundary of the cell. It serves as the critical permeability barrier
consist of the cytoplasmic membrane, cell wall, and, if present, the between the cell and its external environment (see figure 2.30).
capsule (figure 3.23). Bacteria often have capsules, but archaea
rarely do. Enclosed within the envelope are the contents of the cell— Structure of the Cytoplasmic Membrane
the cytoplasm and nucleoid. The cytoplasm is the viscous material The structure of the prokaryotic cytoplasmic membrane is typical
enclosed by the envelope; the fluid portion is called cytosol. The of other biological membranes—a phospholipid bilayer embedded
nucleoid is the gelatinous region where the genetic material resides. with proteins (figure  3.24). The phospholipid molecules are
Unlike the nucleus that characterizes eukaryotic cells, the nucleoid arranged in opposing layers so that their hydrophobic tails face in,
is not enclosed by a membrane. The cell may also have appendages toward the other layer, and their hydrophilic tails face outward,
that give it useful traits, including motility and the ability to adhere interacting freely with aqueous solutions. phospholipids, p. 35
to certain surfaces. The characteristics of typical structures of pro- The proteins embedded in the membrane have a variety of dif-
karyotic cells are summarized in table 3.3. ferent functions. Some act as selective gates, allowing nutrients to

52
 Part I Life and Death of Microorganisms 53

TABLE 3.3 A Summary of Typical Prokaryotic Cell Structures

Structure Characteristics

Extracellular
Filamentous appendages Composed of protein subunits that form a helical chain.
Flagella Provide the most common mechanism of motility.
Pili Different types of pili have different functions. The common types, often called fimbriae, allow cells to adhere to
surfaces. A few types are used for twitching or gliding motility. Sex pili join cells in preparation for DNA transfer.
Capsules and slime layers Layers outside the cell wall, usually made of polysaccharide.
Capsule Distinct and gelatinous. Allows bacteria to adhere to specific surfaces; allows some organisms to evade innate
defense systems and thus cause disease.
Slime layer Diffuse and irregular. Allows bacteria to adhere to specific surfaces.
Cell wall Peptidoglycan provides rigidity to bacterial cell walls, preventing the cells from lysing.
Gram-positive Thick layer of peptidoglycan that contains teichoic acids and lipoteichoic acids.
Gram-negative Thin layer of peptidoglycan surrounded by an outer membrane. The outer layer of the outer membrane is
lipopolysaccharide.
Cell Boundary
Cytoplasmic membrane Phospholipid bilayer embedded with proteins. Surrounds the cytoplasm, separating it from the outside
environment. Also transmits information about the external environment to the inside of the cell.
Intracellular
DNA Contains the genetic information of the cell.
Chromosomal Carries the genetic information required by a cell. Typically a single, circular, double-stranded DNA molecule.
Plasmid Extrachromosomal DNA molecule. Generally carries only genetic information that may be advantageous to a cell in
certain situations.
Endospore A type of dormant cell. Generally extraordinarily resistant to heat, desiccation, ultraviolet light, and toxic
chemicals.
Cytoskeleton Involved in cell division and control of cell shape.
Gas vesicles Small, rigid structures that provide buoyancy to a cell.
Granules Accumulations of high-molecular-weight polymers, synthesized from a nutrient available in relative excess.
Ribosomes Involved in protein synthesis. Two subunits, 30S and 50S, join to form the 70S ribosome.

enter the cell and waste products to exit. Others serve as sensors of
environmental conditions, providing the cell with a mechanism to
monitor and adjust to its surroundings. So, while the cytoplasmic
membrane functions as a permeability barrier, it also transmits
information about the external environment to the inside of cell. Phospholipid
bilayer
protein function, p. 25
Membrane proteins are not stationary; rather, they constantly
drift in the lipid bilayer. Such movement is necessary for the mem-
brane functions. This structure, with its resulting dynamic nature,
is called the fluid mosaic model.
Members of the Bacteria and Archaea have the same general Hydrophilic head
structure of their cytoplasmic membranes, but the lipid composi-
tions are distinctly different. The lipid tails of the archaeal mem-
Hydrophobic tail
brane lipids are connected to glycerol by a different type of chemical
linkage and are not fatty acids. glyerol, p. 33 fatty acid, p. 33
Proteins

MicroByte FIGURE 3.24 The Structure of the Cytoplasmic Membrane

Two opposing layers make up the phospholipid bilayer. Various
E.  coli’s DNA encodes approximately 1,000 different membrane proteins are embedded within the bilayer.
proteins, a large portion of which are involved in transport.
? Which part of the membrane is hydrophobic?
54 Chapter 3 Microscopy and Cell Structure

Permeability of the Lipid Bilayer Water flows across a

The cytoplasmic membrane is selectively permeable, mean- membrane toward the
hypertonic solution.
ing that only certain substances can cross. Those that pass
through the lipid bilayer freely include gases such as O2, CO2,
Hypotonic solution Hypertonic solution
and N2, small hydrophobic molecules, and water (figure 3.25).
Some cells facilitate water passage with aquaporins, pore- Water flow
forming membrane proteins that specifically allow water mol- Solute
ecules to pass through. Other molecules that enter or exit cells molecule
must be moved across the membrane by the transport systems
discussed later.

Simple Diffusion
Molecules that can pass through the lipid bilayer move in and out Water flow
of the cell by simple diffusion, in which molecules move from a
region of high concentration to one of low concentration, until Water flows in Water flows out
equilibrium is reached. The speed and direction of movement
depend on the relative concentration of molecules on each side of
the membrane—the greater the difference in concentration, the
higher the rate of diffusion. The molecules continue to pass
through at a diminishing rate until their concentration is the same
on both sides of the membrane.
Cytoplasmic membrane is Cytoplasmic membrane
forced against cell wall. pulls away from cell wall.
Osmosis (a) (b)
Osmosis is the diffusion of water across a selectively permeable
membrane. It occurs when the concentrations of solute (dissolved
FIGURE 3.26 Osmosis (a) The effect of a hypotonic solution
molecules and ions) on two sides of a membrane are unequal. on a bacterial cell. (b) The effect of a hypertonic solution on a
Typical of diffusion, water moves down its concentration gradient bacterial cell.
from high water concentration (low solute concentration) to low
? What might happen in part (a) if the cell wall were weakened?
water concentration (high solute concentration).
When describing osmosis, three terms are used to refer to the and isotonic (iso means the same). Water flows from the hypo-
solutions on opposing sides of a membrane: hypotonic (hypo tonic solution to the hypertonic one (figure  3.26). No net water
means less; tonic refers to solute), hypertonic (hyper means more), movement occurs between isotonic solutions.

Pass through easily: Passes through: Do not pass through:

Gases (O2, CO2, N2) Water Sugars
Small hydrophobic Ions
molecules Amino acids
ATP Water
Macromolecules

Aquaporin

(a) The cytoplasmic membrane is selectively permeable. Gases, small (b) Aquaporins allow water to pass through the cytoplasmic membrane
hydrophobic molecules, and water are the only substances that more easily.
pass freely through the phospholipid bilayer.
FIGURE 3.25 Permeability of the Lipid Bilayer (a) Selective permeability. (b) The role of aquaporins.
? Why do small charged molecules not pass through the bilayer?
 Part I Life and Death of Microorganisms 55

Osmosis has important biological consequences. The cyto- which is analogous to the energy stored in a battery. electron
plasm of a cell is a concentrated solution of inorganic salts, sugars, transport chain, p. 142
amino acids, and various other molecules. However, the environ- The energy of a proton motive force can be harvested when
ments in which prokaryotes normally grow are typically very dilute protons are allowed to move back into the cell. This is used to
(hypotonic). Water moves toward the high solute concentration, so drive certain cellular processes, including ATP synthesis. It is also
it flows from the surrounding medium into the cell (figure 3.26b). used to power one of the transport systems discussed next and
This inflow of water exerts tremendous osmotic pressure on the some forms of motility.
cytoplasmic membrane, much more than it generally can resist.
However, the strong cell wall surrounding the membrane generally
withstands such high pressure. The cytoplasmic membrane is forced MicroAssessment 3.4
against the wall but cannot balloon further. Damage to the cell wall The cytoplasmic membrane is a phospholipid bilayer embedded with
weakens the structure, and consequently, cells may lyse (burst). proteins. It serves as a selectively permeable barrier between the cell
and the surrounding environment, allowing relatively few substances
to pass through freely. Molecules that pass through the lipid bilayer
move in and out by simple diffusion. Osmosis is the diffusion of
The Role of the Cytoplasmic Membrane water across the membrane. The electron transport chain within the
in Energy Transformation membrane expels protons, generating a proton motive force that is
used to synthesize ATP, and power transport systems and some types
The cytoplasmic membrane of prokaryotic cells plays a critical of motility.
role in transforming energy—converting the energy of food or
9. Explain the fluid mosaic model.
sunlight into ATP, the energy currency of a cell. This is an impor-
10. Name three molecules that pass freely through the lipid bilayer.
tant distinction between prokaryotic and eukaryotic cells; in
eukaryotic cells, this process occurs in membrane-bound organ- 11. Why do the protons ejected by the electron transport system stay
close to the membrane, rather than float away? +
elles, which will be discussed later in this chapter. ATP, p. 24
As part of their energy-transforming processes, most pro-
karyotes have a series of protein complexes, the electron
transport chain, embedded in their membrane. These sequen-
tially transfer electrons and, in the process, move protons out of 3.5 ■ Directed Movement
the cell. The details of the process will be explained in chapter
6. The expulsion of protons by the electron transport chain cre-
of Molecules Across the
ates a proton gradient across the cell membrane—positively Cytoplasmic Membrane
charged protons are concentrated immediately outside the mem-
brane, whereas negatively charged hydroxyl ions remain inside Learning Outcomes
(figure 3.27). The charged ions attract each other, so they stay 10. Explain why transport systems are necessary for a cell.
close to the membrane. This separation of protons and hydroxyl 11. Compare and contrast facilitated diffusion, active transport, and
ions creates an electrochemical gradient across the membrane; group translocation.
inherent in it is a form of energy called proton motive force, 12. Describe the importance of secretion, and explain how a cell
determines which polypeptides are destined for secretion.

Most molecules that enter or exit a cell must pass via proteins
that function as selective gates. This is necessary because the
H+
lipid bilayer of the cytoplasmic membrane prevents nearly all
H+ H+ H+
H+ H+ H+ H+ substances from passing through. Cells must constantly bring
H+ H+ H+ nutrients in through these gates, and expel waste products. They
H+ H+
also need to move certain proteins they synthesize to the outside
of the cell.
Electron transport chain

Transport Systems
OH – OH– OH– OH– Mechanisms that allow nutrients and other small molecules to
OH– OH– OH– OH–
H+ OH–– H+ enter the cell are called transport systems. These are also used to
expel wastes and compounds such as antibiotics and disinfectants
FIGURE 3.27 Proton Motive Force The electron transport chain, that are otherwise damaging to the cell.
a series of protein complexes within the membrane, ejects protons Transported molecules enter the cell through transport proteins,
from the cell. sometimes called permeases or carriers. These span the membrane, so
? Why is proton motive force a form of energy? that one end projects into the surrounding environment and the other
56 Chapter 3 Microscopy and Cell Structure

Small molecule

1 Transport protein recognizes 2 Binding of that molecule changes 3 The molecule is released on the
a specific molecule. the shape of the transport protein. other side of the membrane.

FIGURE 3.28 Transport Protein

? What types of molecules do prokaryotic cells bring in?

into the cell (figure  3.28). The interaction between the transport Active Transport
protein and the molecule it carries is highly specific. Consequently, a Active transport moves compounds against a concentration gra-
single carrier generally transports only a specific type of molecule. dient and requires energy. The two main mechanisms use different
The mechanisms of transport are summarized in table 3.4. forms of energy.
Facilitated Diffusion Transport Systems That Use Proton Motive Force Many pro-
Facilitated diffusion is a form of passive transport, meaning that karyotic transport systems move small molecules and ions in or
it does not require energy. It uses a transport protein to move sub- out of the cell using the energy of a proton motive force (figure
stances from one side of the membrane to the other, but it can only 3.29b). Transporters of this type allow a proton into the cell and
eliminate a difference in concentration, not create one (figure simultaneously either bring along or expel another substance. For
3.29a). Molecules are transported until their concentration is the example, the permease that transports lactose brings the sugar into
same on both sides of the membrane. Because prokaryotes typi- the cell along with a proton. Expulsion of waste products, on the
cally grow in relatively nutrient-poor environments, they gener- other hand, relies on transporters that eject the compound as a
ally cannot rely on facilitated diffusion to take in nutrients. proton passes in. Efflux pumps, which are used by some bacteria
to oust antimicrobial drugs, use this latter mechanism.
Transport Systems That Use ATP Transport mechanisms called
A Summary of Transport ABC transport systems require ATP as an energy source (ABC
TABLE 3.4 Mechanisms Used by stands for ATP Binding-Cassette) (figure  3.29b). These systems
Prokaryotic Cells use specific binding proteins that reside immediately outside of
the cytoplasmic membrane to gather and deliver molecules to the
Transport respective transport complexes.
Mechanism Characteristics

Facilitated Diffusion Rarely used by prokaryotes. Exploits Group Translocation

a concentration gradient to move Group translocation is a transport process that chemically alters
molecules; can only eliminate a a molecule during its passage through the cytoplasmic membrane
gradient, not create one. No energy is
expended.
(figure  3.29c). Typically, this is done by adding a phosphate
group—a process called phosphorylation. Glucose and several
Active Transport Energy is expended to accumulate
molecules against a concentration
other sugars are phosphorylated during their transport. Although
gradient. energy is expended in the process, it can be regained when the
Transporters that As a proton is allowed into the cell
sugar is later broken down, a process described in chapter 6.
use proton motive another substance is either brought
force along or expelled. Protein Secretion
ABC transporters ATP is used as an energy source. Cells actively move certain proteins they synthesize out of the
Extracellular binding proteins deliver a
cell—a process called secretion (figure 3.30). Some of these mol-
molecule to the transporter.
ecules must be moved to the outside because they will become
Group Translocation The transported molecule is chemically
enzymes that break down macromolecules into their individual
altered as it passes into the cell.
subunits. The macromolecules are too large to transport into the
 Part I Life and Death of Microorganisms 57

Transported H+ Binding
substance protein
H+
H+ H+

P~P~P R P
ATP P
H+
R
P~P + Pi
P P
ADP

(a) Facilitated diffusion (b) Active transport, using Active transport, using ATP as an (c) Group translocation
proton motive force as an energy source. A binding protein
Transporter allows a substance energy source. gathers the transported molecules. Transporter chemically
to move across the membrane, alters the substance
but only down the concentration Transporter uses energy (ATP or proton motive force) to move a sub- as it is transported
gradient. stance across the membrane and against a concentration gradient. across the membrane.

FIGURE 3.29 Types of Transport Systems (a) Facilitated diffusion. (b) Active transport. (c) Group translocation.
? Why is facilitated diffusion relatively uncommon in prokaryotes?

cell, but the subunits are not. Others make up external structures MicroAssessment 3.5
such as flagella, the appendages used for motility.
Facilitated diffusion does not require energy. Some active transport
How does the cell machinery know which polypeptides are to
systems use proton motive force as an energy source and others use
be secreted? Those destined for secretion have a characteristic ATP. Group translocation chemically modifies a molecule as it enters
sequence of amino acids, typically at one end of the molecule. the cell, usually by phosphorylation. Proteins destined for secretion
This sequence—a signal sequence—functions as a tag that directs have a characteristic signal sequence.
the secretion machinery to move the preprotein (precursor protein) 12. Prokaryotes rarely rely on facilitated diffusion. Why is this so?
across the membrane. During the transport process, the signal 13. Why would a cell need to secrete proteins?
sequence is removed and the protein ultimately folds into its func-
14. Can you argue that group translocation is a form of
tional shape. Secretion is a complex process, and a variety of dif- active transport? +
ferent secretion systems are used by prokaryotes.

Macromolecule

Extracellular
enzyme
Subunit of
Signal macromolecule
sequence

P~P~P
Preprotein ATP
P~ P + Pi
ADP

a The signal sequence on the preprotein targets b Extracellular enzymes degrade macromolecules
it for secretion and is removed during the so that the subunits can then be transported
secretion process. Once outside the cell, the into the cell using the mechanisms shown in
protein folds into its functional shape. figure 3.29.

FIGURE 3.30 Protein Secretion (a) Generalized view of the protein secretion process. (b) One function of a secreted protein.
? Why would a cell secrete enzymes rather than bring intact macromolecules into the cell?
58 Chapter 3 Microscopy and Cell Structure

3.6 ■ Cell Wall above the peptidoglycan layer and bind cations such as Mg2+.
Their function is not well understood, but they may serve as a
Learning Outcomes reservoir for cations that are essential for enzyme function. They
13. Describe the chemistry and structure of peptidoglycan.
also seem to be important for cell wall construction and cell divi-
sion. Some teichoic acids are covalently joined to the peptido-
14. Compare and contrast the structure and chemistry of the Gram-
positive and Gram-negative cell walls.
glycan molecule, and these are called wall teichoic acids. Others
are linked to the cytoplasmic membrane, and these are called
15. Explain the significance of lipid A and the O antigen of LPS.
lipoteichoic acids.
16. Explain how the cell wall affects susceptibility to penicillin and Recent studies indicate that a gel-like substance is sand-
lysozyme.
wiched between the cytoplasmic membrane and the peptidogly-
17. Explain how the cell wall affects Gram staining characteristics. can layer. This substance is thought to have a function similar to
18. Describe the cell walls of the Archaea. the periplasm associated with Gram-negative cell walls, dis-
cussed next.
The prokaryotic cell wall is a strong, somewhat rigid structure that
prevents the cell from bursting. Its architecture distinguishes two
main groups of bacteria—Gram-positive and Gram-negative
(table 3.5). Comparison of Features
TABLE 3.5 of Gram-Positive and
Gram-Negative Bacteria
Peptidoglycan
Outer membrane
The strength of the Gram-positive and Gram-negative bacterial Peptidoglycan Gel-like
and teichoic acids material Periplasm
cell walls is due to a layer of peptidoglycan, a material found only
in bacteria (figure 3.31). The basic structure of peptidoglycan is
an alternating series of two major subunits: N-acetylmuramic acid
(NAM) and N-acetylglucosamine (NAG). These subunits, which Cytoplasmic Peptidoglycan
are related to glucose, are covalently joined to one another to form membrane
Cytoplasmic membrane
a glycan chain. This linear polymer serves as the backbone of the
peptidoglycan molecule. Gram-Positive Gram-Negative
Attached to each of the NAM molecules is a tetrapeptide
Color of Gram- Purple Pink
chain (a string of four amino acids) that plays an important role in Stained Cell
the strength of peptidoglycan. The tetrapeptide chains connect
together, linking adjacent glycan chains to form a single, very Representative Bacillus, Escherichia,
Genera Staphylococcus, Neisseria,
large three-dimensional molecule, much like a flexible, multi-
Streptococcus Pseudomonas
layered chain-linked fence. In Gram-negative bacteria, tetrapep-
tides are joined directly. In Gram-positive bacteria, they are Distinguishing Structures/Components
usually linked indirectly by a peptide interbridge (a series of Peptidoglycan Thick layer Thin layer
amino acids).
Only a few types of amino acids make up the tetrapeptide Teichoic acids Present Absent
chains. One of these, diaminopimelic acid (related to lysine), is Outer membrane Absent Present
not found in any other place in nature. Some of the others are
d-stereoisomers, a form found in relatively few substances. Lipopolysaccharide Absent Present
(endotoxin)
lysine, p. 26 D-stereoisomer, p. 27
Porin proteins Absent Present; allow
(unnecessary molecules to pass
The Gram-Positive Cell Wall because there through outer
is no outer membrane
A relatively thick layer of peptidoglycan characterizes the Gram- membrane)
positive cell wall (figure 3.32). As many as 30 layers of intercon-
nected glycan chains make up the polymer. Regardless of its General Characteristics
thickness, peptidoglycan is permeable to sugars, amino acids, and Sensitivity to Generally more Generally less
many other substances. penicillin susceptible susceptible
In addition to peptidoglycan, the Gram-positive cell wall (with notable (with notable
has teichoic acids (from the Greek word teichos, meaning wall). exceptions) exceptions)
These are negatively charged chains of a common subunit (either Sensitivity to Yes No
ribitol-phosphate or glycerol-phosphate) to which various sugars lysozyme
and d-alanine are typically attached. Teichoic acids stick out
 Part I Life and Death of Microorganisms 59

N-acetylmuramic acid N-acetylglucosamine

(NAM) (NAG)

CH2OH CH2OH
O O
H H Chemical structure of N-acetylglucosamine (NAG)
O O O and N-acetylmuramic acid (NAM); the ring structure
H H OH H H of each molecule is glucose.
H NH H NH
O
C O C O
HC CH3
CH3 CH3
C O
OH

Glycan
NAG NAM NAG NAM
chain

Peptidoglycan
Peptide interbridge
(Gram-positive cells)
Tetrapeptide
chain Glycan chains are composed of
(amino acids) alternating subunits of NAG and
NAM. They are cross-linked via
their tetrapeptide chains to
create peptidoglycan.

NAM NAG NAM NAG

NAM NAG
Glycan
chain

Interconnected glycan chains

form a large sheet. Multiple
connected layers create a
three-dimensional molecule.

Tetrapeptide chain Peptide interbridge FIGURE 3.31 Components and Structure of

(amino acids) Peptidoglycan
? Why is peptidoglycan medically important?
60 Chapter 3 Microscopy and Cell Structure

N-acetylglucosamine N-acetylmuramic acid Teichoic acid

Peptidoglycan Gel-like
and teichoic acids material

Peptidoglycan
(cell wall)

Cytoplasmic
membrane

Gel-like Gram-positive
material (b)

Cytoplasmic
membrane
Cytoplasmic
Peptidoglycan membrane

(a)

FIGURE 3.32 Gram-Positive Cell Wall (a) The Gram-positive cell wall has a
relatively thick layer of peptidoglycan consisting of many sheets of interconnected
glycan chains. (b) Simple diagram of a cross-section of the structure. (c) Gram-positive
(c) 0.15 µm
cell wall TEM (Bacillus subtilis).
? What connects the glycan chains in peptidoglycan?

The Gram-Negative Cell Wall Two parts of the LPS molecule are particularly notable
(figure 3.33b):
The Gram-negative cell wall contains only a thin layer of peptido-
glycan (figure  3.33). Outside of that is the outer membrane, a ■ Lipid A anchors the LPS molecule in the lipid bilayer. This
unique lipid bilayer embedded with proteins. The outer membrane is the portion of the LPS molecule that the body recognizes as
is joined to peptidoglycan by lipoproteins. lipoproteins, p. 29 the sign of invading Gram-negative bacteria.
■ O antigen is the portion of LPS directed away from the mem-
The Outer Membrane brane, at the end opposite lipid A. It is made up of a chain of
The outer membrane of Gram-negative bacteria is unique. Its sugar molecules, the number and type of which varies among
bilayer structure is typical of other membranes, but the outside different species. The differences can be used to identify cer-
layer is made up of a molecule called lipopolysaccharide (LPS) tain species or strains.
rather than phospholipid. LPS is extremely important from a
medical standpoint. When injected into an animal, it causes symp- Like the cytoplasmic membrane—which in Gram-negative
toms characteristic of infections caused by live bacteria. The bacteria is sometimes called the inner membrane—the outer
symptoms are actually the body’s response to LPS, which serves membrane serves as a barrier to the passage of most molecules.
as a signal to the defense systems that Gram-negative bacteria It excludes many compounds that could damage the cell,
have invaded. If very small quantities of LPS enter the tissues, including certain antimicrobial medications. This is one reason
such as when a few bacterial cells contaminate a minor wound, the why Gram-negative bacteria are generally less sensitive to
defense systems respond at a level that can safely eliminate the many such medications. Small molecules and ions can cross
invader. When large amounts of the molecule accumulate, how- the membrane through porins, specialized channel-forming
ever, such as when Gram-negative bacteria are actively growing proteins that span the outer membrane. Some porins are spe-
in the bloodstream, the magnitude of the response can be deadly. cific for certain molecules; others allow many different mole-
Because of this lethal effect, LPS is called endotoxin (endo mean- cules to pass.
ing that it is inside the cell, although it is actually a component of Gram-negative bacteria have a number of unique secretion
the envelope). endotoxin, p. 394 systems that move proteins across both the cytoplasmic and outer
 Part I Life and Death of Microorganisms 61

O antigen
(varies in length and
Porin protein composition)

Core polysaccharide

Lipid A

Lipopolysaccharide (b)
(LPS) Outer
membrane
(lipid bilayer)
Outer Peptidoglycan
membrane
Lipoprotein

Periplasm

Periplasm
Cytoplasmic
membrane
Peptidoglycan
(c)

Cytoplasmic
membrane
(inner membrane;
lipid bilayer)

Outer Cytoplasmic
Peptidoglycan membrane Periplasm membrane

(a)

(d) 0.15 µm

FIGURE 3.33 Gram-Negative Cell Wall (a) The Gram-negative cell wall has a thin layer of peptidoglycan made up of only one or two sheets
of interconnected glycan chains. The outer membrane is a typical phospholipid bilayer, except the outer layer is lipopolysaccharide. Porins span
the membrane to allow specific molecules to pass. Periplasm fills the region between the two membranes. (b) Structure of lipopolysaccharide. The
lipid A portion is responsible for the symptoms associated with endotoxin. The sugars in the O antigen vary among bacterial species. (c) Simple
diagram of a cross-section of the structure. (d) Gram-negative cell wall TEM (Pseudomonas aeruginosa).
? Why is lipopolysaccharide medically significant?

membranes. These play a critical role in the disease process of the periplasm unless specifically moved across the outer mem-
some pathogens, so medical microbiologists are very interested in brane as well. Thus, periplasm is filled with proteins involved in a
learning more about how they function. One hope is that medica- variety of cellular activities, including nutrient degradation and
tions can be developed to jam these systems. type III secretion transport. For example, the enzymes that cells export to break
systems, p. 386 down peptides and other molecules are in periplasm. Similarly,
the binding proteins of the ABC transport systems are found there.
Periplasm
The region between the cytoplasmic membrane and the outer
MicroByte
membrane is the periplasmic space, which is filled with a gel-like
The “O157” in E. coli O157:H7 refers to the characteristic O antigen.
substance called periplasm. All exported proteins accumulate in
62 Chapter 3 Microscopy and Cell Structure

Antibacterial Substances
That Target Peptidoglycan
Compounds that interfere with the synthesis of peptidoglycan or alter
its structural integrity weaken the molecule to a point where it can no
longer prevent the cell from bursting. These substances have no effect
on eukaryotic cells because peptidoglycan is unique to bacteria.
Penicillin
Penicillin is the most thoroughly studied of a group of antibiotics
that interfere with peptidoglycan synthesis. It functions by prevent-
ing the cross-linking of adjacent glycan chains. penicillin, p. 463
Generally, but with notable exceptions, penicillin is far more
effective against Gram-positive bacteria than Gram-negative ones.
This is because the outer membrane of Gram-negative cells pre-
vents the medication from reaching the peptidoglycan layer. 2 µm
Scientists have developed derivatives of penicillin that cross the FIGURE 3.34 Mycoplasma pneumoniae These cells vary in
outer membrane, however, and these can be effective in treating shape because they lack a cell wall.
infections caused by Gram-negative bacteria. ? Would lysozyme or penicillin affect M. pneumoniae?

Lysozyme
of flat protein or glycoprotein subunits. These subunits self-assemble,
Lysozyme—an enzyme found in tears, saliva, and many other body
so they may have applications in nanotechnology—a branch of sci-
fluids—breaks the bonds that link the alternating subunits of the gly-
ence that seeks to build functional items from molecules and atoms.
can chain. This destroys the structural integrity of the peptidoglycan
Some bacterial cells have S-layers as well, but they are in addition to
molecule. Lysozyme is sometimes used in the laboratory to remove
a cell wall, rather than serving as the primary structural component.
the peptidoglycan layer from bacteria for experimental purposes.

Cell Wall Type and the Gram Stain MicroAssessment 3.6

The type of bacterial cell wall accounts for the Gram stain reac- Peptidoglycan is a molecule unique to bacteria that provides strength
to the cell wall. The Gram-positive cell wall is composed of a
tion, but it is the inside of the cell, not the wall, that is stained by
relatively thick layer of peptidoglycan as well as teichoic acids.
crystal violet. Gram-positive cells retain the dye because their cell Gram-negative cell walls have a thin layer of peptidoglycan and a
wall prevents the crystal violet–iodine complex from being lipopolysaccharide-containing outer membrane. The outer membrane
washed out by the decolorizing agent, whereas Gram-negative excludes many molecules. Penicillin and lysozyme interfere with the
cells lose the dye quite easily. The decolorizing agent is thought structural integrity of peptidoglycan. Mycoplasma species lack a cell
to dehydrate the thick layer of peptidoglycan, and in this desic- wall. Members of the Archaea have a variety of cell wall types.
cated state the wall acts as a permeability barrier—the barrier 15. What is the significance of lipid A?
prevents the dye from leaving the cell. In contrast, the solvent 16. How does the action of penicillin differ from that of lysozyme?
action of the decolorizing agent easily damages the outer mem- 17. Explain why penicillin kills only actively multiplying cells,
brane of Gram-negative bacteria, and the relatively thin layer of whereas lysozyme kills cells in any stage of growth. +
peptidoglycan cannot retain the dye complex. Gram stain, p. 47

Bacteria That Lack a Cell Wall 3.7 ■ Capsules and Slime Layers
Some bacteria naturally lack a cell wall. Mycoplasma species, one
of which causes a mild form of pneumonia, have an extremely Learning Outcome
variable shape because they lack a rigid cell wall (figure 3.34). As 19. Compare and contrast the structure and function of capsules and
expected, neither penicillin nor lysozyme affects these organisms. slime layers.
Mycoplasma and related bacteria can survive without a cell wall
because their cytoplasmic membrane has sterols in it, making it Many bacteria have a gel-like layer outside the cell wall that either
stronger than that of most other bacteria. sterol, p. 36 protects the cell or allows it to attach to a surface (figure 3.35). If
the layer is distinct and gelatinous, it is a capsule. If, instead, the
Cell Walls of the Domain Archaea layer is diffuse and irregular, it is a slime layer. Colonies that
As a group, members of the Archaea have a variety of cell wall types. form either of these often appear moist and glistening.
This is probably due to the fact that they inhabit a wide range of Capsules and slime layers vary in their chemical composition,
environments, including some that are extreme. The Archaea have depending on the microbial species. Most are composed of poly-
not been studied as extensively as the Bacteria, however, so less is saccharides, and are commonly referred to as a glycocalyx (glyco
known about the structure of their walls. None of the Archaea have means “sugar” and calyx means “shell”). A few capsules consist
peptidoglycan in their cell wall, but some do have a similar molecule of polypeptides made up of repeating subunits of only one or two
called pseudopeptidoglycan. Many have S-layers, which are sheets amino acids. Interestingly, the amino acids are generally of the
 Part I Life and Death of Microorganisms 63

MicroAssessment 3.7
Cell in intestine Capsules and slime layers allow organisms to adhere to surfaces and
sometimes protect bacteria from the host defense systems.
18. How do capsules differ from slime layers?
19. Explain why a sugary diet can lead to tooth decay. +

3.8 ■ Filamentous Protein

Appendages
Capsule Learning Outcomes
20. Describe the structure and arrangements of flagella, and explain
how they are involved in chemotaxis.
21. Compare and contrast the structure and function of fimbriae and
sex pili.

Many prokaryotes have protein appendages anchored in the cyto-

(a) 2 µm
plasmic membrane that protrude out from the surface. These
structures are not essential to the life of microbes, but they do give
the cells a competitive advantage.

Flagella
Flagella (singular: flagellum) are long protein structures respon-
sible for most types of prokaryotic motility (figure 3.36). By spin-
ning like propellers, flagella push a cell through liquid much as a

(b) 1 µm

FIGURE 3.35 Capsules and Slime Layers (a) Encapsulated bacte-

ria attaching to intestinal cells (EM). (b) Masses of bacteria adhering in
a layer of slime (SEM).
? What is the function of capsules and slime layers?

d-stereoisomeric form, one of the few places these are found in

nature. polysaccharide, p. 31 D-amino acid, p. 27
(a) 1 µm
Some capsules and slime layers allow bacterial cells to adhere
to specific surfaces, including teeth, rocks, and other bacteria.
Once attached, the cells can grow as a biofilm, a polymer-encased
community of microbes. One example is dental plaque, a biofilm
on teeth. When a person ingests sucrose, Streptococcus mutans (a
common member of the oral microbiota) can use that to make a
glycocalyx. This allows S. mutans to attach to other bacterial cells
on the teeth. In turn, additional bacteria adhere to the sticky gly-
cocalyx, resulting is an even greater accumulation of plaque.
Acids produced by bacteria in dental plaque then damage the tooth
surface. biofilm, p. 84 dental plaque, p. 575
Some capsules allow bacteria to avoid host defense systems
that otherwise protect against infection. Streptococcus pneumoniae,
(b) 1 µm
an organism that causes bacterial pneumonia, can cause the disease
only if it has a capsule. Unencapsulated cells are quickly engulfed FIGURE 3.36 Flagella (a) Peritrichous flagella (SEM). (b) Polar
and killed by phagocytes, an important cell of the body’s defense flagellum (SEM).
system. Streptococcus pneumoniae, p. 497 phagocytes, p. 346 ? How can flagella affect a microbe’s ability to cause disease?
64 Chapter 3 Microscopy and Cell Structure

ship is driven through water. Flagella must work very hard to at one end of the cell. Other arrangements include a tuft of fla-
move a cell, because water has the same relative viscosity to pro- gella at one or both ends of a cell (see figure 3.18).
karyotes as molasses has to humans.
In some cases, flagella are important in disease. For example, Chemotaxis
Helicobacter pylori, the bacterium that causes gastric ulcers, has Motile bacteria sense the presence of chemicals and respond by
powerful multiple flagella at one end of the cell. These flagella moving in a certain direction—a phenomenon called chemotaxis.
allow H. pylori to penetrate the viscous mucous gel that coats the If a compound is a nutrient, it may serve as an attractant, causing
stomach epithelium. Helicobacter pylori, p. 578 cells to move toward it. On the other hand, if the compound is
toxic, it may act as a repellent, causing cells to move away.
MicroByte The movement of a bacterial cell toward an attractant is not
Flagella can rotate over 100,000 rpm, propelling a cell 20 body lengths
a second. This is equivalent to a 6-foot-tall man running 82 mph!
in a straight line (figure  3.38). When an E.  coli cell travels, it
progresses in one direction for a short time, but then stops and
tumbles for a fraction of a second. As a consequence, the cell usu-
Structure and Arrangement of Flagella ally finds itself oriented in a completely different direction. It then
A flagellum has three basic parts—a filament, hook, and basal body moves in that direction for a short time, and tumbles again. The
(figure 3.37). The filament is the portion that extends into the exter- seemingly odd pattern of travel is due to the coordinated rotation
nal environment. It is made up of identical subunits of a protein of the flagella. When flagella rotate counterclockwise, they form
called flagellin. These subunits form a chain that twists into a helical a tight propelling bundle and the bacterium is pushed in a forward
structure with a hollow core. The hook, a flexible curved segment, movement called a run. After a brief period, the direction of rota-
connects the filament to the cell surface. The basal body anchors the tion of the flagella reverses. This change causes the cell to stop
structure to the cell wall and cytoplasmic membrane. and tumble. When cells sense movement toward an attractant they
While the flagella of bacteria and archaea share the same tumble less frequently, so the runs are longer. Cells also tumble
general structure, they have many differences. For instance, bacte- less frequently when they are moving away from a repellent.
rial flagella are powered by proton motive force, whereas archaeal In addition to reacting to chemicals, some bacteria respond to the
flagella use ATP for energy. The molecules that compose archaeal concentration of O2 (aerotaxis). Organisms that require O2 for growth
flagella and the mechanisms of their assembly are also will move toward it, whereas bacteria that grow
distinct. only in its absence tend to be repelled by it.
The numbers and arrangement of flagella Certain motile bacteria can react to the
can be used to characterize flagellated bacte- earth’s magnetic field by the process of
ria. As an example, E.  coli have flagella magnetotaxis. They actually contain a
Flagellin
distributed over the entire surface, an row of magnetic particles that cause
arrangement called peritrichous (peri Filament
the cells to line up in a north-to-south
means “around”). Other common bacteria direction much as a compass does
have a polar flagellum, a single flagellum (figure 3.39). The magnetic forces of

Hook

Flagellum

FIGURE 3.37 The

Structure of a Flagellum
in a Gram-Negative
Bacterium The flagellum
is composed of three basic
parts—a filament, a flexible
hook, and a basal body.
? What is the role of
flagellin? E. coli

Basal
body

Harvests the energy

of the proton motive force
to rotate the flagellum.
 Part I Life and Death of Microorganisms 65

A cell moves via a series of runs and tumbles.

Tumble (T) Run (R) Tumble (T)

Sex pilus

Flagellum
The cell moves randomly When a cell senses it is moving toward
when there is no an attractant, it tumbles (T) less frequently, Other pili
concentration gradient of resulting in longer runs (R).
attractant or repellent.

T Gradient of attractant concentration

T T
(a) 1 µm

R Epithelial cell
R

FIGURE 3.38 Chemotaxis

Bacterium
? What mechanism causes a cell to tumble?

the earth attract the organisms so that they move downward and into Bacterium
sediments where the O2 concentration is low—the environment best with pili
suited for their growth. Some bacteria can move in response to varia-
tions in temperature (thermotaxis) or light (phototaxis).

Pili
Pili are considerably shorter and thinner than flagella, and their
function is quite different. However, one part of their structure has (b) 5 µm
a theme similar to the filament of a flagellum—a string of protein FIGURE 3.40 Pili (a) Pili on an Escherichia coli cell. The short pili
subunits arranged helically to form a long molecule with a hollow (fimbriae) mediate adherence; the sex pilus is involved in DNA transfer.
core (figure 3.40). (b) Escherichia coli attaching to epithelial cells in the small intestine of
Many types of pili allow cells to attach to specific surfaces; a pig.
these pili are also called fimbriae. Strains of E.  coli that cause ? How does the structure and function of pili compare to that of
watery diarrhea have pili that allow them to attach to cells that line flagella?
the small intestine. Without the ability to attach, these cells would
simply be propelled through the intestinal tract along with the
other intestinal contents. enterotoxigenic E. coli, p. 590
Some types of pili help bacterial cells move on solid media.
Twitching motility (characterized by jerking movements) and
Flagellum certain types of smooth, gliding motility involve pili.
Another type of pilus, called a sex pilus, is used to join one
bacterium to another for a specific type of DNA transfer. This and
other mechanisms of DNA transfer will be described in chapter 8.
DNA transfer, p. 200

MicroAssessment 3.8
Magnetite particles
Flagella are the most common mechanism for bacterial motility. Chemo-
0.4 mm taxis is the directed movement of cells toward an attractant or away from
FIGURE 3.39 Magnetotactic Bacterium The chain of magnetic a repellent. Pili provide a mechanism for attachment to specific surfaces.
particles (magnetite: Fe3O4) within Magnetospirillum magnetotacti- 20. E. coli cells have peritrichous flagella. What does this mean?
cum helps align the cell along geomagnetic lines (TEM).
21. How are fimbriae different from sex pili in their function? +
? Why would magnetotaxis benefit a cell?
66 Chapter 3 Microscopy and Cell Structure

3.9 ■ Internal Structures these medications. Because a bacterium can sometimes transfer a
copy of a plasmid to another bacterial cell, this accessory genetic
Learning Outcomes information can spread, which accounts in large part for the
22. Describe the structure and function of the chromosome, plasmids,
increasing frequency of antibiotic-resistant organisms.
ribosomes, storage granules, gas vesicles, and endospores.
23. Describe the significance and processes of sporulation and Ribosomes
germination.
Ribosomes are involved in protein synthesis, where they serve as
Prokaryotic cells have a variety of structures within the cell. Some, the structures that facilitate the joining of amino acids. Their rela-
such as the chromosome and ribosomes, are essential for the life of tive size and density is expressed as a distinct unit, S (for
all cells, whereas others give cells certain selective advantages. Svedberg), that reflects how fast they settle when spun at very
high speeds in an ultracentrifuge. The faster they move toward the
bottom, the higher the S value and the greater the density.
The Chromosome Prokaryotic ribosomes are 70S. Note that S units are not strictly
The prokaryotic chromosome is typically a single, circular arithmetic; the 70S ribosome is composed of a 30S and a 50S
double-stranded DNA molecule that contains all the genetic infor- subunit (figure 3.42). Prokaryotic ribosomes differ from eukary-
mation required by a cell. It occurs as a mass within the cyto- otic ribosomes, which are 80S. The fact that they are distinct is
plasm, forming a gel-like region called the nucleoid. important medically because antibiotics that interfere with the
Chromosomal DNA is tightly packed into the cell function of the 70S ribosome have no effect on the 80S molecule.
(figure  3.41). The compact shape is due partially to nucleoid- function of ribosomes, p. 171

associated proteins that bind to DNA, creating a structure that

bends and folds. In addition, the DNA is twisted, or supercoiled.
This can be visualized by cutting a rubber band and then twisting
Cytoskeleton
one end several times before rejoining cut the ends. The resulting It was once thought that bacteria lacked a cytoskeleton, an interior
circle will twist and coil in response. protein framework. Several bacterial proteins that have similari-
ties to those of the eukaryotic cytoskeleton have now been charac-
terized, and these appear to be involved in cell division and
Plasmids controlling cell shape.
Most plasmids are circular, supercoiled, double-stranded DNA
molecules. They are generally much smaller than the chromosome
and carry from a few to several hundred genes. A single cell can Storage Granules
have more than one type of plasmid, and these can each be present Storage granules are accumulations of high-molecular-weight
in multiple copies. plasmids, p. 208 polymers synthesized from a nutrient that a cell has in relative
A cell generally does not require the genetic information car- excess. If nitrogen and/or phosphorus are lacking, for example, a
ried by a plasmid. However, the encoded characteristics can be cell cannot multiply even if a carbon and energy source such as
advantageous in certain situations. For example, many plasmids glucose is plentiful. Rather than waste the carbon/energy source,
code for the production of enzymes that destroy certain antibiot- cells use it to produce glycogen, a glucose polymer. Later, when
ics, allowing the organism to resist the otherwise lethal effect of conditions are appropriate, cells degrade and use the glycogen.

DNA

(a) 0.5 µm (b) 1.3 µm

FIGURE 3.41 The Chromosome (a) Color-enhanced transmission electron micrograph of a thin section of Escherichia coli, with the DNA
shown in red. (b) Chromosome released from a gently lysed E. coli cell. Note how tightly packed the DNA must be inside the bacterium.
? What is the gel-like region formed by the chromosome called?
 Part I Life and Death of Microorganisms 67

Gas Vesicles
Some aquatic bacteria produce gas vesicles—small, rigid, protein-
50S subunit bound compartments that provide buoyancy to the cell. Gases, but
not water, flow freely into the vesicles, thereby decreasing the
density of the cell. By regulating the number of gas vesicles, a cell
can float or sink to its ideal position in the water column. Bacteria
30S subunit that use sunlight as a source of energy use gas vesicles to float
closer to the surface, where light is available.

Endospores
An endospore is a unique type of dormant cell produced by cer-
30S + 50S tain bacterial species such as members of the genera Bacillus and
combined Clostridium (figure 3.44). The structures may remain dormant for
perhaps 100 years, or even longer, and are extraordinarily resistant
to damaging conditions including heat, desiccation, toxic chemi-
cals, and ultraviolet (UV) light. Immersion in boiling water for
hours may not kill them. Endospores that survive these treatments
70S ribosome can germinate, or exit the dormant stage, to become a typical
multiplying cell, called a vegetative cell.
FIGURE 3.42 70S Ribosome The 70S ribosome is composed of 50S
and 30S subunits. Because endospores can survive so long in a variety of condi-
tions, they can be found virtually everywhere. They are common
? What is the function of ribosomes?
in soil, which can make its way into environments such as labora-
tories and hospitals and onto products such as medical devices,
Other bacterial species store carbon and energy as poly-β- food, and media used to cultivate microbes. Excluding microbes
hydroxybutyrate (figure 3.43). from these environments and products is very important, so spe-
Some types of granules can be readily detected by light cial precautions must be taken to avoid or destroy endospores.
microscopy. Volutin granules, a storage form of phosphate, stain Endospores are sometimes called spores. However, this latter
red with the dye methylene blue, whereas the surrounding cellular term is also used to refer to structures produced by unrelated
material stains blue. Because of this, they are often called meta- microbes such as fungi. Bacterial endospores are much more resis-
chromatic granules (meta means “change” and chromatic means tant to environmental conditions than are other types of spores.
“color”). Recent evidence suggests that the role of these granules
MicroByte
is more complex than originally thought. The volutin granules of The diseases botulism, tetanus, gas gangrene, and anthrax are all
some bacteria are membrane-bound and appear to resemble caused by endospore-formers.
eukaryotic organelles that are thought to be involved with energy
storage and pH balance.
Sporulation
MicroByte Endospore formation, or sporulation, is a complex sequence
Poly-β-hydroxybutyrate can be used to make biodegradable plastics. of changes that begin when spore-forming bacteria experience
Phosphate-storing cells can be used to remove pollutants from
wastewater.

Storage granules 0.5 µm

Endospore 1 µm
FIGURE 3.43 Storage Granules The large unstained areas in the 
photosynthetic bacterium Rhodospirillum rubrum are granules of poly-β- FIGURE 3.44 Endospores Endospore inside a vegetative cell of a
hydroxybutyrate. Clostridium species (TEM).
? How would storage granules benefit a cell? ? What is the function of an endospore?
68 Chapter 3 Microscopy and Cell Structure

limiting amounts of carbon or nitrogen. The cells sense starvation

1
conditions, which triggers them to begin the 8-hour process. Vegetative growth stops;
After vegetative growth stops, the DNA is duplicated and a DNA is duplicated.
septum forms, dividing the cell asymmetrically (figure 3.45). The
larger compartment then engulfs the smaller compartment, form-
ing a forespore within a mother cell. These two portions take on
different roles in synthesizing the components that will make up
the endospore. The forespore will ultimately become the core of
the endospore. Peptidoglycan-containing material is laid down 2
A septum forms, dividing
between the two membranes that surround the forespore, forming the cell asymmetrically.
the core wall and the cortex. Meanwhile, the mother cell makes
proteins that will form the spore coat. Ultimately, the mother cell
is degraded and the endospore released.
The layers of the endospore protect it from damage. The spore
coat is thought to function as a sieve, excluding molecules such as
lysozyme. The cortex helps maintain the core in a dehydrated
3
state, protecting it from the effects of heat. In addition, the core The larger compartment
has small, acid-soluble proteins that bind to DNA, thereby protect- engulfs the smaller
ing it from damage. The core is rich in an unusual compound compartment, forming a
forespore within a
called calcium dipicolinate, which seems to also play an important mother cell.
role in spore resistance.

Germination
Germination can be triggered by a brief exposure to heat or cer- 4 Forespore
Peptidoglycan-containing
tain chemicals. Following such exposure, the endospore takes on material is laid down between
water and swells. The spore coat and cortex then crack open, and the two membranes that now
a vegetative cell grows out. Since one vegetative cell gives rise to surround the forespore.
one endospore, sporulation is not a means of cell reproduction.

MicroAssessment 3.9 Peptidoglycan-containing Mother cell

material
The prokaryotic chromosome is usually a circular, double-stranded DNA
molecule that contains all of the genetic information required by a cell.
Plasmids generally encode only information that is advantageous to a Core wall
5
cell in certain conditions. Ribosomes are the structures that facilitate the The mother cell is degraded
joining of amino acids to form a protein. The cytoskeleton is an interior and the endospore released.
framework involved in cell division and controlling cell shape. Storage
granules are polymers synthesized from nutrients a cell has in relative
excess. Gas vesicles provide buoyancy to a cell. An endospore is a
highly resistant dormant stage produced by certain bacterial species.
22. Explain how glycogen granules benefit a cell.
23. Explain why endospores are an important concern for the Cortex Spore coat
canning industry.
24. Why are the processes of sporulation and germination not a
mechanism of multiplication? + FIGURE 3.45 The Process of Sporulation
? Approximately how long does the sporulation process take?

THE EUKARYOTIC CELL

Eukaryotic cells are generally much larger than prokaryotic cells, information (DNA). The organelles, which can take up half the
and their internal structures are far more complex (figure 3.46). total cell volume, allow the cell to perform complex functions in
One of their distinguishing characteristics is the abundance of separated regions. For example, when degradative enzymes are
membrane-enclosed compartments or organelles. The most contained within an organelle, they can digest material without
important of these is the nucleus, which contains the cell’s genetic posing a threat to the integrity of the cell itself.
 Part I Life and Death of Microorganisms 69

Nucleus
A vesicle forms when a The mobile vesicle can then move to
Nuclear envelope Cytoplasm section of an organelle other parts of the cell, ultimately fusing
Nucleolus buds off. with the membrane of another organelle.
Rough
Plasma membrane endoplasmic
reticulum Protein
Centriole
with ribosomes

Mitochondrion Smooth
endoplasmic
Cytoskeleton reticulum
Actin filament Migrating Fusing
Ribosomes Budding
Microtubule transport vesicle
Golgi vesicle
Intermediate vesicle
filament apparatus
Peroxisome
(a) Lysosome
FIGURE 3.47 Vesicle Formation and Fusion
Rough Nucleus ? The lumen is which part of an organelle?
endoplasmic Nuclear envelope
reticulum Nucleolus
with ribosomes Cytoskeleton
Intermediate
Smooth filament
endoplasmic
reticulum Microtubule
Actin
Ribosomes filament
Each organelle contains a variety of compounds, many of
Golgi
apparatus Central which are synthesized at other locations. To deliver these to the
vacuole Peroxisome lumen (interior) of another organelle, a section of the organelle
Mitochondrion buds off, forming a membrane-bound vesicle (figure 3.47). This
Chloroplast
(opened to
carries a bit of the organelle’s contents to other parts of the cell.
show thylakoids) When the vesicle encounters another organelle, the two mem-
branes fuse to become one unit, similar to two oil droplets merg-
Adjacent cell wall ing. By doing so, the vesicle spills its contents into the organelle.
As a group, eukaryotic cells are highly variable. For exam-
Cell wall
Cytoplasm ple, protozoa, which are single-celled organisms, must function
Plasma membrane
(b)
exclusively as self-contained units that seek and ingest food.
These cells must be mobile and flexible to take in food particles,
so they lack cell walls that would otherwise provide rigidity.
Animal cells also lack a cell wall, because they too must be flex-
ible to accommodate movement. Fungi, on the other hand, are
Cell
stationary and benefit from the protection provided by a strong
membrane cell wall. Fungal cell walls are composed primarily of polysac-
charides including chitin, a polymer of N-acetylglucosamine.
Plants, which are also stationary, have cell walls composed of
Nucleus
cellulose, a polymer of glucose. polysaccharides, p. 31 chitin,
Nuclear p. 32 cellulose, p. 32
membrane The individual cells of a multicellular organism can be dis-
tinctly different from one another. Liver cells, for example, are
obviously quite different from bone cells. Different cell types,
Mitochondrion however, can function in cooperative associations called tissues.
The tissues in your body include muscle, connective, nerve, epi-
thelial, blood, and lymphoid. Each of these provides a unique
function. Various tissues work together to make up organs,
(c) 1 µm including skin, heart, and liver. Organs and the tissues that com-
pose them will be covered in more detail in chapters on infectious
FIGURE 3.46 Eukaryotic Cells (a) Diagrammatic representation
of an animal cell. (b) Diagrammatic representation of a plant cell. diseases.
(c) Micrograph of an animal cell shows several membrane-bound struc- A comprehensive coverage of all aspects of eukaryotic cells
tures including mitochondria and a nucleus. Note that a prokaryotic is beyond the scope of this textbook. Instead, this section will
cell is about the size of a mitochondrion. focus on key characteristics, particularly those that directly affect
? Which organelle contains the cell’s genetic information? the interactions of microbe with human hosts. These characteristics
70 Chapter 3 Microscopy and Cell Structure

TABLE 3.6 A Summary of Typical Eukaryotic Cell Structures

Structure Characteristics

Plasma Membrane Asymmetric lipid bilayer embedded with proteins. Permeability barrier, transport, and cell-to-cell
communication.
Internal Protein Structures
Cilia Beat in synchrony to provide movement. Composed of microtubules in a 9 + 2 arrangement.
Cytoskeleton Dynamic filamentous network that provides structure to the cell.
Flagella Propel or push the cell with a whiplike or thrashing motion. Composed of microtubules in a 9 + 2
arrangement.
Ribosomes Two subunits, 60S and 40S, join to form the 80S ribosome.
Membrane-Bound Organelles
Chloroplasts Site of photosynthesis; the organelle harvests the energy of sunlight to generate ATP, which is then
used to convert CO2 to carbohydrates.
Endoplasmic reticulum Site of synthesis of macromolecules destined for other organelles or the external environment.
Rough Attached ribosomes thread proteins they are synthesizing into the lumen of the organelle.
Smooth Site of lipid synthesis and degradation, and calcium ion storage.
Golgi apparatus Site where macromolecules synthesized in the endoplasmic reticulum are modified before being
transported in vesicles to other destinations.
Lysosome Digestion of macromolecules.
Mitochondria Harvest the energy released during the degradation of organic compounds to generate ATP.
Nucleus Contains the genetic information (DNA).
Peroxisome Site where oxidation of lipids and toxic chemicals occurs.

are summarized in table  3.6. The functions of prokaryotic and structures, helping to maintain cell integrity. Those that face the
eukaryotic cell components are compared in table 3.7. outside often function as receptors. A given receptor binds a
specific molecule, which is referred to as its ligand. These
receptor-ligand interactions are extremely important in multicel-
3.10 ■ The Plasma Membrane lular organisms because they allow cells to communicate with
each other—a process called signaling. For example, when a cell
Learning Outcome within the human body encounters a compound unique to bac-
24. Describe the structure and function of the eukaryotic plasma teria, it secretes specific proteins as a call for help. Other cells
membrane, comparing and contrasting it with the prokaryotic have receptors for these proteins on their surface, allowing them
cytoplasmic membrane. to recognize the signals, and respond accordingly. Using this cell-
to-cell communication, a multicellular organism can function as
All eukaryotic cells have a cytoplasmic membrane, or plasma a cohesive unit.
membrane, which is similar in chemical structure and function to The membranes of many eukaryotic cells contain sterols,
that of prokaryotic cells. It is a typical phospholipid bilayer which provide strength to the otherwise fluid structure. The sterol
embedded with proteins. The lipid and protein composition of the in animal cell membranes is cholesterol, whereas fungal mem-
layer that faces the cytoplasm, however, differs significantly from branes have ergosterol. This difference is exploited by antifungal
that facing the outside of the cell. The same is true for membranes medications that act by interfering with ergosterol synthesis or
that surround the organelles. The layer facing the inside of the function. antifungal medications, p. 476
organelle is similar to its plasma membrane counterpart that faces Within the plasma membrane are cholesterol-rich regions
the outside of the cell. This lack of symmetry reflects the impor- called lipid rafts. The role of these regions is still being studied,
tant role these membranes play in complex processes within the but they appear to be important in allowing the cell to detect and
eukaryotic cell. respond to signals in the external environment. From a microbi-
The membrane proteins perform a variety of functions. Some ologist’s perspective, they are also important because many
are involved in transport and others are attached to internal viruses use these regions when they enter or exit a cell.
 Part I Life and Death of Microorganisms 71

TABLE 3.7 Comparison of Typical Prokaryotic and Eukaryotic Cell Structures/Functions

Prokaryotic Eukaryotic

General Characteristics
Size Generally 0.3–2 μm in diameter. Generally 5–50 μm in diameter.
Cell division Chromosome replication followed by binary Chromosome replication and mitosis followed by
fission. division.
Chromosome location Located in the nucleoid, which is not Contained within the membrane-bound nucleus.
membrane-bound.
Structures
Cell membrane Relatively symmetric with respect to the lipid Highly asymmetric; lipid composition of outer layer
content of the bilayers. differs significantly from that of inner layer.
Cell wall Composed of peptidoglycan (Bacteria); Absent in animal cells; composition in other cell
Gram-negative bacteria have an outer types may include chitin, glucans and mannans
membrane as well. (fungi), and cellulose (plants).
Chromosome Single, circular DNA molecule is typical. Multiple, linear DNA molecules. DNA is wrapped
around histones.
Flagella Composed of protein subunits; attached Made up of a 9 + 2 arrangement of microtubules;
to the cell envelope. covered by an extension of the plasma membrane.
Membrane-bound organelles Absent. Present; includes the nucleus, mitochondria,
chloroplasts (only in plant cells), endoplasmic
reticulum, Golgi apparatus, lysosomes, and
peroxisomes.
Nucleus Absent; DNA resides as an irregular mass forming Present.
the nucleoid region.
Ribosomes 70S ribosomes, which are made up of 50S and 80S ribosomes, which are made up of 60S and 40S
30S subunits. subunits. Mitochondria and chloroplasts have 70S
ribosomes.
Functions
Degradation of extracellular Enzymes are secreted that degrade Macromolecules can be brought into the cell by
substances macromolecules outside of the cell. The resulting endocytosis. Lysosomes carry digestive enzymes.
small molecules are transported into the cell.
Motility Generally involves flagella, which are composed Involves cilia and flagella, which are made up of a
of protein subunits. Flagella rotate like 9 + 2 arrangement of microtubules. Cilia move in
propellers. synchrony; flagella propel a cell with a whiplike
motion or thrash back and forth to pull a cell
forward.
Protein secretion Secretion systems transport proteins across the Secreted proteins are moved to the lumen of the
cytoplasmic membrane. rough endoplasmic reticulum as they are being
synthesized. From there, they are transported to the
Golgi apparatus for processing and packaging.
Strength and rigidity Peptidoglycan-containing cell wall (Bacteria). Cytoskeleton composed of microtubules, intermediate
filaments, and microfilaments. Some have a cell wall;
some have sterols in the membrane.
Transport Primarily active transport. Group translocation. Facilitated diffusion and active transport. Ion
channels.

The plasma membrane plays no role in ATP synthesis; MicroAssessment 3.10

instead, that task is performed by mitochondria (discussed later). The plasma membrane is an asymmetric lipid bilayer embedded
Even though the plasma membrane has no electron transport chain with proteins. Specific receptors allow cell-to-cell signaling. Sterols
to generate a proton motive force, there is still an electrochemical provide strength to the fluid membrane.
gradient across the membrane. This is maintained by energy- 25. What is the medical significance of ergosterol in fungal membranes?
consuming mechanisms that pump either sodium ions or protons
26. Describe why signaling is important in animal cells.
to the outside of the cell. electrochemical gradient, p. 55
27. How could one argue that the interior of an organelle is
“outside” of a cell? +
72 Chapter 3 Microscopy and Cell Structure

3.11 ■ Transfer of Molecules example is an ABC transporter used by cancerous cells that ejects
therapeutic drugs intended to kill those cells.
Across the Plasma
Membrane Endocytosis and Exocytosis
Learning Outcomes Endocytosis is the process by which a eukaryotic cell takes up
25. Compare and contrast the roles of channels and carriers. material from the surrounding environment by forming invagina-
26. Describe the processes of endocytosis and exocytosis. tions (inward folds) in its membrane (figure  3.48). The type of
27. Describe the role of the endoplasmic reticulum in secretion.
endocytosis common to most animal cells is pinocytosis. In this
process, the small invaginations carry along some liquid and any
Various substances—including nutrients, signaling molecules, material attached to the membrane. This action ultimately forms a
and waste products—must pass through the plasma membrane. membrane-bound compartment called an endosome, which then
Cells have several mechanisms to accomplish this. fuses with digestive organelles called lysosomes to form an
endolysosome. The characteristics of lysosomes will be discussed
shortly. lysosome, p. 77
Animal cells often take up material by receptor-mediated
Transport Proteins endocytosis, a variation of pinocytosis. It allows cells to take up
The transport proteins of eukaryotic cells function as either carri- extracellular ligands that bind to receptors on the cell’s surface.
ers or channels. Carriers are analogous to proteins in prokaryotic When the receptors bind their ligands, the region is internalized to
cells that function in facilitated diffusion and active transport. form an endosome that contains the receptors along with their
Channels form small pores in the membrane that allow only spe- bound ligands. The acidic internal environment of the endosome
cific ions to diffuse through. To control ion passage, the channel frees the ligands from the receptors, which are then often recycled.
has a gate that can open or close in response to environmental After that, the endosome fuses with lysosomes.
conditions. Eukaryotic cells also have aquaporins. facilitated dif- Protozoa and phagocytes, both of which ingest bacteria and
fusion, p. 56 active transport, p. 56 aquaporin, p. 54 large debris, use a specific type of endocytosis called phagocytosis.
Cells of multicellular organisms often take up nutrients by Phagocytes are important cells of the body’s defense system
facilitated diffusion. This is possible because systems can regulate against microbial invaders. In phagocytosis, the cells send out
the nutrient concentration surrounding the cells. For instance, when armlike extensions called pseudopods, which surround and
the human body maintains a high glucose concentration in the enclose extracellular material, including bacteria. This action
blood, tissue cells can bring the sugar in without using energy. brings the material into the cell in an enclosed compartment called
The active transport mechanisms of eukaryotic cells are struc- a phagosome. This fuses with endosomes, and ultimately fuses
turally similar to those of prokaryotic cells. A medically important with lysosomes to form a phagolysosome. phagocytes, p. 346

Endocytosis Fusion with lysosomes Exocytosis

Pinocytosis— Lysosome
cells take in liquids.

Digestive
Receptor- Endosome enzymes Endolysosome
mediated Nucleus
endocytosis—
cells take in
material that
has bound
to receptors. Endosome Exocytic
Endolysosome vesicle

Phagosome
Phagocytosis— Phagolysosome
cells engulf
Exocytosis
particulate material Pseudopod
such as bacteria.

FIGURE 3.48 Endocytosis and Exocytosis

? How is pinocytosis different from phagocytosis?
 Part I Life and Death of Microorganisms 73

The process of exocytosis is the reverse of endocytosis.

Membrane-bound exocytic vesicles inside the cell fuse with the
plasma membrane and release their contents to the outside.

MicroByte
Many viruses use receptor-mediated endocytosis to enter animal cells. Microtubule
They bind to a specific receptor, “tricking” the cell into bringing them in.

Secretion
As in prokaryotic cells, proteins destined for secretion have a sig- Intermediate filament
nal sequence, a characteristic amino acid sequence that functions Actin filament
as a tag. When ribosomes synthesize a protein with such a
sequence, they attach to a complex on the membrane of the endo- Plasma membrane
plasmic reticulum (ER)—an organelle that will be described
shortly. As the protein is made, it is threaded through the mem-
brane and into the ER. Once inside the ER, the protein can easily
be transported by vesicles to the outside of the cell. Proteins des-
(a) Actin filament
tined for various organelles also have specific amino acid tags.
 endoplasmic reticulum, p. 76

MicroAssessment 3.11
Transport proteins function as either channels or carriers. Pinocytosis
allows cells to internalize small molecules. Protozoa and phagocytes
internalize bacteria and debris by phagocytosis. Exocytosis is used to (b) Microtubule
expel material. Secreted proteins are threaded through the membrane
of the endoplasmic reticulum as they are being made.
28. How does a cell bring in ligands?
29. How is the formation of an endosome different from that of a
phagosome?
30. How might a bacterium resist the killing effects of a
phagolysosome? +
(c) Intermediate filament
FIGURE 3.49 Cytoskeleton Diagrammatic representation of the
dynamic filamentous network that provides structure to the cell; the
3.12 ■ Protein Structures cytoskeleton is composed of three elements: (a) actin filaments,
(b) microtubules, and (c) intermediate filaments.
Within the Cell ? What is the role of actin filaments?

Learning Outcome
28. Describe the structure and function of eukaryotic ribosomes, the Actin filaments allow the cell cytoplasm to move. The fila-
cytoskeleton, flagella, and cilia. ments are polymers of protein subunits called actin, which rapidly
assemble and subsequently disassemble to cause motion. For
Eukaryotic cells have a variety of important protein structures example, pseudopod formation relies on actin polymerization in
within the cell. These include ribosomes, the cytoskeleton, fla- one part of the cell and depolymerization in another.
gella, and cilia. Microtubules, the thickest of the cytoskeleton’s components,
are long hollow structures made of protein subunits called tubulin.
Ribosomes Microtubules form the mitotic spindles, the machinery that sepa-
rates duplicated chromosomes as cells divide. Without mitotic
Ribosomes are the structures involved in protein synthesis. The
spindles, cells could not reproduce. In addition, microtubules are
eukaryotic ribosome is 80S, and is made up of a 60S and a 40S
the main structures that make up the cilia and flagella, the mecha-
subunit. Recall that the prokaryotic ribosomes are 70S.
nisms of locomotion in some eukaryotic cells. Microtubules also
function as the framework along which organelles and vesicles
Cytoskeleton move within a cell. The antifungal drug griseofulvin interferes
The cytoskeleton forms the framework of a cell (figure 3.49). Its with the action of microtubules in some fungi. mitosis, p. 283
three components—(1) actin filaments (also called microfilaments), Intermediate filaments function like ropes, strengthening
(2) microtubules, and (3) intermediate filaments—continually the cell mechanically. By providing mechanical support, they
reconstruct to adapt to the cell’s constantly changing needs. allow cells to resist physical stresses.
74 Chapter 3 Microscopy and Cell Structure

Outer MicroAssessment 3.12

microtubule pair
The 80S eukaryotic ribosome is composed of 60S and 40S subunits.
The cytoskeleton is a dynamic filamentous network that provides
structure to the cell; it is composed of actin filaments, microtubules,
Flagellum and intermediate filaments. Flagella function in motility. Cilia either
propel a cell or move material along a stationary cell.
31. Explain how actin filaments are related to phagocytosis.
32. Explain what is meant by the 9 + 2 structure of cilia and
flagella.
Central
Plasma microtubule pair
33. Why would 60S and 40S ribosomal subunits make an 80S
membrane ribosome rather than a 100S ribosome? +
Basal body

Microtubule 3.13 ■ Membrane-Bound

triplet
Organelles
Learning Outcome
29. Describe the function of the nucleus, mitochondria, chloroplasts,
endoplasmic reticulum, Golgi apparatus, lysosomes, and
peroxisomes.
FIGURE 3.50 Flagellum A flexible structure involved in movement.
Membrane-bound organelles are an important feature of eukary-
? How is the structure of a eukaryotic flagellum different from its
prokaryotic counterpart?
otic cells that sets them apart from prokaryotic cells.

The Nucleus
An important distinguishing feature of a eukaryotic cell is the
nucleus, which contains the genetic information. The boundary of
MicroByte
this structure is the nuclear envelope, composed of two lipid bi-
Some intracellular pathogens trigger actin polymerization, which
propels the pathogens with enough force to push them into an layer membranes—the inner and outer membranes (figure 3.51).
adjacent cell. Complex protein structures span the envelope, forming nuclear
pores. These allow large molecules such as ribosomal subunits
and proteins to be transported into and out of the nucleus. The
nucleolus is a region where ribosomal RNAs are synthesized.
Flagella and Cilia The nucleus contains multiple chromosomes, each one encod-
ing different genetic information. Unlike the situation in most
Flagella and cilia are flexible structures that appear to project out
prokaryotic cells, the DNA is linear. It is wound around proteins
of a cell yet are covered by extensions of the plasma membrane
called histones, a characteristic that adds structure and order to the
(figure 3.50). They are composed of long microtubules grouped in
long molecule.
what is called a 9 + 2 arrangement—nine pairs of microtubules
Events that take place in the nucleus during cell division dis-
surrounding two individual ones. They originate from basal bodies
tinguish eukaryotes from prokaryotes. After DNA is replicated in
within the cell; the basal body has a slightly different arrangement
eukaryotic cells, chromosomes go through a nuclear division pro-
of microtubules.
cess called mitosis, which ensures that the daughter cells receive
Although eukaryotic flagella function in motility, they are
the same number of chromosomes as the original parent. Because
structurally very different from their prokaryotic counterparts.
of mitosis, a cell that is diploid (has two copies of each chromo-
Using ATP as a source of energy, they either propel the cell
some) will generate two diploid daughter cells. A different pro-
with a whiplike motion or thrash back and forth to pull the cell
cess, meiosis, generates haploid daughter cells, which each have a
forward.
single copy of each chromosome.
Cilia are shorter than flagella, often covering a cell and mov-
ing in synchrony (see figure 1.8). Their motion can move a cell
forward in an aqueous solution, or propel surrounding material Mitochondria
along a stationary cell. For example, epithelial cells that line the Mitochondria function as ATP-generating powerhouses, and are
respiratory tract have cilia that beat together in a directed fashion. found in nearly all eukaryotic cells. They are complex structures
This moves the mucus film that covers those cells, directing it bounded by two lipid bilayers—the outer and inner membranes
toward the mouth, where it can be swallowed. Because of this (figure  3.52). The outer membrane is smooth, but the inner
action, called the mucociliary escalator, most microbes that have membrane is highly folded, forming invaginations called cristae.
been inhaled are removed before they can enter the lungs. The folds increase the membrane’s surface area, maximizing the
 Part I Life and Death of Microorganisms 75

Nuclear
pores

Nuclear
envelope

Nucleolus
Nucleus

Nuclear
pores

Vacuole
Inner membrane

Outer membrane

Nuclear pore

(a) (b) 0.3 µm

FIGURE 3.51 Nucleus Organelle that contains the cell’s genetic information (DNA). (a) Diagrammatic representation. (b) Electron micrograph
of a yeast cell by freeze-fracture technique.
? What is the function of nuclear pores?

Ribosome

Matrix
DNA
Crista

Intermembrane space
Inner membrane
Outer membrane

(a) (b) 0.1 µm

FIGURE 3.52 Mitochondria Organelles that harvest energy released during the degradation of organic compounds to synthesize ATP.
(a) Diagrammatic representation. (b) Electron micrograph.
? What were the first pieces of evidence that led scientists to conclude that mitochondria evolved from bacterial cells?

ATP-generating capabilities of the organelle (the processes will be cytoplasm of eukaryotic cells. This observation, and the fact that
discussed in chapter 6). mitochondria divide in a fashion similar to that of bacteria, were
The mitochondrial matrix—the gel-like material enclosed by among the first pieces of evidence that led scientists to hypothesize
the mitochondrial inner membrane—contains DNA, ribosomes, that mitochondria evolved from bacterial cells. This evidence was
and other molecules necessary for protein synthesis. Notably, the bolstered by DNA sequencing data, supporting what is now known
ribosomes are 70S rather than the 80S ribosome found in the as the endosymbiotic theory (see Perspective 3.1).
76 Chapter 3 Microscopy and Cell Structure

PERSPECTIVE 3.1
The Origins of Mitochondria and Chloroplasts
Mitochondria and chloroplasts bear such a for some of the ribosomal proteins and ribo- allows scientists to easily determine the nucleo-
striking similarity to prokaryotic cells that it somal RNAs that make up their 70S ribosomes. tide sequence of DNA molecules, making it
is no wonder scientists speculated for many These ribosomes contrast with the typical 80S possible to compare organelle DNA with
decades that these organelles evolved from ribosomes that characterize eukaryotic cells genomes of different bacteria. This led to the
bacteria. The endosymbiont theory states that and, in fact, are equivalent to the prokaryotic discovery that some mitochondrial DNA
the ancestors of mitochondria and chloroplasts 70S ribosomes. Interestingly, nuclear DNA sequences bear a striking resemblance to DNA
were bacteria residing within other cells in a encodes some of the components that make up sequences of members of a group of obligate
mutually beneficial partnership. The intracel- these ribosomes. Another characteristic that intracellular parasites, the rickettsias. These are
lular bacterium in such a partnership is called supports the theory that mitochondria and chlo- probably relatives of modern-day mitochondria.
an endosymbiont. As time went on each roplasts were once intracellular bacteria is the A tremendous effort is now underway to
partner became indispensable to the other, and double membrane that surrounds these organ- determine the nucleotide sequence of mito-
the endosymbiont eventually lost key features elles. Present-day endosymbionts retain their chondria from a wide variety of eukaryotes,
such as a cell wall and the ability to replicate cytoplasmic membranes and live within mem- including plants, animals, and protists. While
independently. brane-bound compartments in their eukaryotic the size of mitochondrial DNA varies a great
Several early observations have supported host cell. In addition, mitochondria and chloro- deal among these different eukaryotic organ-
the endosymbiont theory. Mitochondria and plasts multiply by elongating and then dividing isms, common sequence themes are emerging.
chloroplasts, unlike other eukaryotic organelles, (binary fission), just as bacteria do. Today, researchers are no longer discussing
both carry some of the genetic information nec- Evidence in favor of the endosymbiont the- “if” but “when” these organelles evolved from
essary for their function. These include genes ory continues to accumulate. Recent technology intracellular prokaryotes.

Chloroplasts As with mitochondria, substantial evidence indicates that

chloroplasts evolved from bacterial cells (see Perspective 3.1).
Chloroplasts, found exclusively in plants and algae, are the site
Chloroplasts contain DNA and 70S ribosomes, elongate and
of photosynthesis in eukaryotic cells. They harvest the energy of
divide, and have photosynthetic mechanisms and DNA sequences
sunlight to generate ATP, which is then used to convert CO2 to
similar to a group of bacteria called cyanobacteria.
sugar and starch. Like mitochondria, chloroplasts are bound by
two membranes (figure 3.53). Within the chloroplast’s stroma, the
substance analogous to the mitochondrial matrix, are membrane- Endoplasmic Reticulum (ER)
bound, disclike structures called thylakoids. Chlorophyll and other The endoplasmic reticulum (ER) is a complex system of flattened
pigments that capture radiant energy are embedded in the thyla- sheets, sacs, and tubes (figure  3.54). The rough endoplasmic
koid membranes. reticulum has a characteristic bumpy appearance due to the

Ribosome DNA Thylakoids

Stroma

Thylakoid membrane
Outer membrane
Inner membrane
Thylakoid disc

FIGURE 3.53 Chloroplasts These harvest the energy of sunlight to generate ATP, which is then used to convert CO2 to an organic form.
? Chloroplasts evolved from which group of bacteria?
 Part I Life and Death of Microorganisms 77

Rough
endoplasmic
reticulum

Ribosomes

Smooth
endoplasmic
reticulum

FIGURE 3.54 Endoplasmic Reticulum Site of synthesis of macromolecules destined for other
organelles or the external environment.
? What causes the bumpy appearance of the rough endoplasmic reticulum? 0.08 µm

many ribosomes adhering to the surface. It is the site where pro- Lysosomes and Peroxisomes
teins not destined for the cytoplasm are synthesized. These
Lysosomes are organelles that contain a number of powerful deg-
include proteins targeted for secretion or transfer to an organ-
radative enzymes that could destroy the cell if not contained
elle’s lumen. Membrane proteins such as receptors are also syn-
within the organelle. Endosomes and phagosomes fuse with lyso-
thesized on the rough ER. When ribosomes begin making these
somes, so that material taken up by the cell can be degraded. In a
proteins, they attach to the ER surface so that the polypeptides
similar manner, old organelles or vesicles formed around unneeded
they are synthesizing can be threaded through gated pores in the
cellular components can fuse with lysosomes. This process,
membrane. This delivers the molecules to the ER’s lumen,
autophagy, digests the cell’s own contents.
where they then fold to assume their three-dimensional shapes.
Peroxisomes are the organelles in which O2 is used to help
Vesicles that bud off from the ER then transfer the newly syn-
break down lipids and detoxify certain chemicals. The organelle’s
thesized proteins to the Golgi apparatus for further modification
enzymes generate highly reactive molecules such as hydrogen
and sorting. polypeptide, p. 27
peroxide and superoxide. The peroxisome contains these mole-
Some regions of the ER are smooth. This smooth endoplasmic
cules and ultimately degrades them, protecting the cell from their
reticulum functions in lipid synthesis and degradation, and calcium
toxic effects. hydrogen peroxide, p. 90 superoxide, p. 90
ion storage. As with material made in the rough ER, vesicles trans-
fer compounds from the smooth ER to the Golgi apparatus.
MicroAssessment 3.13
The nucleus, which contains the genetic information, is a
distinguishing feature of a eukaryotic cell. Mitochondria are ATP-
generating powerhouses. Chloroplasts are the site of photosynthesis.
The Golgi Apparatus The rough endoplasmic reticulum is the site where proteins not
The Golgi apparatus consists of a series of membrane-bound destined for the cytoplasm are synthesized. The smooth ER functions
in lipid synthesis and degradation, and calcium ion storage. The Golgi
flattened compartments (figure 3.55). It is the site where macro- apparatus modifies and sorts molecules synthesized in the rough ER.
molecules synthesized in the endoplasmic reticulum are modified Lysosomes are the organelles within which digestion takes place. In
before transport to other destinations. These modifications, such peroxisomes, O2 is used to break down substances.
as the addition of carbohydrate and phosphate groups, take place 34. Describe the structure of the nucleus.
in a sequential order in different compartments of the Golgi. Much
35. How does the function of the rough endoplasmic reticulum differ
like an assembly line, the molecules are transferred in vesicles from that of the smooth endoplasmic reticulum?
from one compartment to another. The molecules are then sorted
36. Why is it logical that bacterial cells do not contain
and delivered in vesicles to specific cellular compartments or to chloroplasts? +
the outside of the cell.
78 Chapter 3 Microscopy and Cell Structure

Vesicle

Fusing
vesicle
Forming
vesicle

Vesicle

0.57 µm

FIGURE 3.55 Golgi Apparatus Site where macromolecules synthesized in the endoplasmic reticulum are modified before being transported
to other destinations.
? How are the modified macromolecules transported from the Golgi apparatus to other sites?

FUTURE CHALLENGES 3.1

A Case of Breaking and Entering
Unraveling the complex mechanisms that pro- be prevented from bringing in nutrients and A more thorough understanding of
karyotic and eukaryotic cells use to transport removing wastes, their growth would cease. eukaryotic uptake systems could be used to
materials across their membranes can poten- Another strategy would be to enhance the develop better antiviral drugs. Viruses exploit
tially aid in the development of new antimi- uptake or decrease the efflux of a specific the process of receptor-mediated endocytosis
crobial medications. Armed with a precise compound that interferes with intracellular to gain entry into the cell. Once they are
model of the structure and function of bacterial processes. This is already being done to some enclosed within the endosome, their protec-
transporter proteins, scientists might be able to extent as new derivatives of current antibiotics tive protein coat is removed, releasing their
design new drugs that exploit these systems. are being produced, but more precise under- genetic material. New drugs can potentially
One strategy would be to design compounds standing of the processes by which bacteria be developed that block these steps, prevent-
that irreversibly bind to transporter molecules take up or remove compounds could speed ing the entry or uncoating of infectious viral
and jam the mechanism. If the microbes can drug development. particles.

Summary
MICROSCOPY AND CELL MORPHOLOGY (DIC) microscopes cause an image to appear three-dimensional
(figure 3.6).
3.1 ■ Microscopic Techniques: The Instruments (table 3.1)
Other Light Microscopes
Principles of Light Microscopy: The Bright-Field Fluorescence microscopes are used to observe cells stained with fluo-
Microscope rescent dyes (figure 3.7). Scanning laser microscopes (SLMs) are used
The most commonly used type of microscope is the bright-field micro- to obtain interior views of intact cells and three-dimensional images of
scope (figure 3.1). The usefulness of a microscope depends largely on its thick structures (figure 3.8).
resolving power (figure 3.2).
Electron Microscopes
Light Microscopes That Increase Contrast Transmission electron microscopes (TEMs) transmit electrons
Cells viewed through a dark-field microscope stand out against a dark through a specimen (figure  3.10). Scanning electron microscopes
background (figure  3.4). Phase-contrast microscopes increase dif- (SEMs) scan a beam of electrons over the surface of a specimen
ferences in refraction (figure  3.5). Differential interference contrast (figure 3.11).
 Part I Life and Death of Microorganisms 79

Atomic Force Microscopes Protein Secretion

Atomic force microscopes map surfaces on an atomic scale (figure 3.12). A characteristic signal sequence targets proteins for secretion (figure 3.30).

3.2 ■ Microscopic Techniques: Dyes and Staining (table 3.2) 3.6 ■ Cell Wall

Differential Staining Peptidoglycan (figure 3.31)

The Gram stain is widely used: Gram-positive bacteria stain purple Peptidoglycan provides rigidity to the cell wall of bacteria. It is com-
and Gram-negative bacteria stain pink (figure  3.14). The acid-fast posed of glycan strands connected via tetrapeptide chains.
stain is used to stain Mycobacterium species; acid-fast organisms stain The Gram-Positive Cell Wall (figure 3.32)
pink and all other organisms stain blue (figure 3.15). The Gram-positive cell wall contains a relatively thick layer of pepti-
Special Stains to Observe Cell Structures doglycan. Teichoic acids project out of the peptidoglycan. A gel-like
The capsule stain allows the capsule to stand out as a clear zone around substance is sandwiched between the cytoplasmic membrane and pep-
a cell (figure 3.16). The endospore stain uses heat to facilitate the stain- tidoglycan layer.
ing of endospores (figure  3.17). The flagella stain uses a substance The Gram-Negative Cell Wall (figure 3.33)
that allows a stain to adhere to and coat the otherwise thin flagella The Gram-negative cell wall has a relatively thin layer of peptido-
(figure 3.18). glycan. An outer membrane contains lipopolysaccharides. LPS is
Fluorescent Dyes and Tags called endotoxin. Porins permit small molecules to pass through the
outer membrane. Periplasm fills the space between the inner and outer
Some fluorescent dyes bind compounds that characterize all cells;
membranes.
others bind compounds specific to certain cell types (figure  3.19).
Immunofluorescence is used to tag proteins of interest with fluorescent Antibacterial Substances That Target Peptidoglycan
compounds. Penicillin prevents peptidoglycan synthesis. Lysozyme destroys the
structural integrity of peptidoglycan.
3.3 ■ Morphology of Prokaryotic Cells
Cell Wall Type and the Gram Stain
Shapes Gram-positive cells retain the crystal violet–iodine dye complex even
Most prokaryotes are cocci or rods; other common shapes are vibrios, when subjected to decolorization, but Gram-positive cells do not.
spirilla, and spirochetes (figure 3.20).
Bacteria That Lack a Cell Wall
Groupings Mycoplasma species are variable in shape and not affected by lysozyme
Cells adhering to one another following division form characteristic or penicillin (figure 3.34).
arrangements such as chains, packets, and clusters (figure 3.22). Cell Walls of the Domain Archaea
Multicellular Associations Archaea have a greater variety of cell wall types than do the Bacteria,
Myxobacteria move as a pack and form fruiting bodies. Most bacteria and they all lack peptidoglycan.
on surfaces live as biofilms.
3.7 ■ Capsules and Slime Layers
Capsules and slime layers allow bacteria to adhere to surfaces. Some
THE PROKARYOTIC CELL (figure 3.23; table 3.3) capsules allow pathogens to avoid host defense systems (figure 3.35).
3.4 ■ The Cytoplasmic Membrane
3.8 ■ Filamentous Protein Appendages
Structure of the Cytoplasmic Membrane (figure 3.24)
Flagella (figure 3.36)
The cytoplasmic membrane is a phospholipid bilayer embedded with
Flagella are used for motility (figure 3.37). Chemotaxis is movement
a variety of different proteins.
toward an attractant or away from a repellent (figure 3.38). Cells use
Permeability of the Lipid Bilayer phototaxis, aerotaxis, magnetotaxis, and thermotaxis to move toward
The cytoplasmic membrane is selectively permeable. A few com- light, O2, a magnetic field, and temperature, respectively.
pounds pass through by simple diffusion (figure 3.26). Some of the Pili (figure 3.40)
membrane proteins function as selective gates.
Many types of pili (fimbriae) allow specific attachment of cells to sur-
The Role of the Cytoplasmic Membrane in Energy faces. Sex pili are involved in a form of DNA transfer.
Transformation
3.9 ■ Internal Structures
The electron transport chain generates an electrochemical gradient, a
source of energy called proton motive force (figure 3.27). The Chromosome (figure 3.41)
The chromosome forms a region called the nucleoid; it contains the
3.5 ■ Directed Movement of Molecules Across genetic information required by a cell.
the Cytoplasmic Membrane
Plasmids
Transport Systems (figures 3.28, 3.29; table 3.4)
Plasmids encode genetic information that may be helpful to a cell.
Facilitated diffusion moves compounds by exploiting a concentration
gradient. Active transport moves compounds against a concentration Ribosomes (figure 3.42)
gradient and requires energy. Group translocation chemically modi- Ribosomes are involved in protein synthesis. The 70S bacterial ribo-
fies a molecule during its transport. some is composed of a 50S and a 30S subunit.
80 Chapter 3 Microscopy and Cell Structure

Cytoskeleton 3.12 ■ Protein Structures Within the Cell

The cytoskeleton is involved in cell division and regulation of shape. Ribosomes
Storage Granules (figure 3.43) The 80S ribosome is composed of 60S and 40S subunits.
Storage granules are synthesized from nutrients a cell has in excess. Cytoskeleton (figure 3.49)
Gas Vesicles The cytoskeleton is composed of actin filaments, microtubules, and
Gas vesicles provide buoyancy to aquatic cells. intermediate filaments.
Flagella and Cilia (figure 3.50)
Endospores
Flagella propel a cell or pull the cell forward. Cilia move in synchrony
Endospores are resistant to heat, desiccation, toxic chemicals, and UV
to either propel a cell or move material along a stationary cell.
light; they can germinate to become vegetative cells (figures 3.44, 3.45).
3.13 ■ Membrane-Bound Organelles

THE EUKARYOTIC CELL (figure 3.46; table 3.6) The Nucleus (figure 3.51)

The nucleus contains the cell’s genetic information (DNA).
3.10 ■ The Plasma Membrane
The plasma membrane is a phospholipid bilayer embedded with pro- Mitochondria
teins that are involved in transport, structural integrity, and signaling. Mitochondria use the energy released during the degradation of
organic compounds to generate ATP (figure 3.52).

3.11 ■ Transfer of Molecules Across Chloroplasts

the Plasma Membrane Chloroplasts capture the energy of sunlight, and then use it to synthe-
size ATP. This is used to convert CO2 to an organic form (figure 3.53).
Transport Proteins
Carriers function in facilitated diffusion and active transport. Endoplasmic Reticulum (ER) (figure 3.54)
Channels form pores in the membrane. These channels are gated. Proteins not destined for the cytoplasm are synthesized in the rough
endoplasmic reticulum. The smooth endoplasmic reticulum is where
Endocytosis and Exocytosis (figure 3.48)
lipids are synthesized and degraded, and calcium is stored.
Pinocytosis is used to take up liquids. Receptor-mediated endocytosis is
used by animal cells to take up material that binds to receptors. Protozoa The Golgi Apparatus (figure 3.55)
and phagocytes take up material using phagocytosis. Exocytosis expels The Golgi apparatus modifies and sorts molecules synthesized in the
material. endoplasmic reticulum.
Secretion Lysosomes and Peroxisomes
Proteins destined for a secretion are made by ribosomes attached to the Lysosomes carry digestive enzymes. Peroxisomes are the organelles
endoplasmic reticulum. in which O2 is used to oxidize certain substances.

Review Questions
Short Answer b) Confocal scanning microscope
1. Explain why resolving power is important in microscopy. c) Phase-contrast microscope
2. Explain why basic dyes are used more frequently than acidic dyes d) Scanning electron microscope
in staining. e) Transmission electron microscope
3. Describe what happens at each step in the Gram stain. 2. When a medical technologist wants to determine if a clinical
4. Compare and contrast ABC transport systems with group translocation. specimen contains a Mycobacterium species, which should be
5. Give two reasons why the outer membrane of Gram-negative bac- used?
teria is medically significant. a) Acid-fast stain b) Capsule stain c) Endospore stain
6. Compare and contrast penicillin and lysozyme. d) Gram stain e) Simple stain
7. Describe how a plasmid can help a cell. 3. Penicillin
8. How is an organ different from tissue? 1. is generally effective against Gram-positive bacteria.
9. How is receptor-mediated endocytosis different from phagocytosis? 2. is generally effective against Gram-negative bacteria.
10. Explain how the Golgi apparatus cooperatively functions with the 3. functions in the cytoplasm of the cell.
endoplasmic reticulum. 4. is effective against Mycoplasma species.
5. kills only growing cells.
Multiple Choice a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5
1. Which of the following is most likely to be used in a typical micro- 4. Endotoxin is associated with
biology laboratory? a) Gram-positive bacteria. b) Gram-negative bacteria.
a) Bright-field microscope c) the cytoplasmic membrane. d) the endospore.
 Part I Life and Death of Microorganisms 81

5. The “O157” in the name E. coli O157:H7 refers to the type of 2. A research laboratory is investigating environmental factors that
O antigen. From this information you know that E. coli inhibit the growth of archaea. They wonder if penicillin would be
a) has a capsule. b) is a rod. effective in controlling their growth. Explain the probable results
c) is a coccus. d) is Gram-positive. of an experiment in which penicillin is added to a culture of
archaea.
e) is Gram-negative.
6. Eliminating which structure is always deadly to cells? Critical Thinking +
a) Flagella b) Capsule c) Cell wall 1. This graph shows facilitated diffusion of a compound across a
d) Cytoplasmic membrane e) Fimbriae cytoplasmic membrane and into a cell. As the external concentra-
7. Which of the following do bacterial cells use for attachment? tion of the compound is increased, the rate of uptake increases
1. Capsule 2. Pilus 3. Cytoplasmic membrane. until it reaches a point where it slows and then begins to plateau.
This is not the case with passive diffusion, where the rate of uptake
4. Periplasm 5. Peptidoglycan
continually increases. Why does the rate of uptake slow and then
a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5 eventually plateau with facilitated diffusion?
8. Endocytosis is associated with
a) mitochondria. b) prokaryotic cells. c) eukaryotic cells.
d) chloroplasts. e) ribosomes.
9. Protein synthesis is associated with
1. lysosomes. 2. the cytoplasmic membrane.

Solute transport rate

3. the Golgi apparatus. 4. rough endoplasmic reticulum.
5. ribosomes.
a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5
10. If a eukaryotic cell were treated with a chemical that destroys
tubulin, all of the following would be directly affected except
a) actin. b) cilia. c) eukaryotic flagella.
d) microtubules. e) More than one of these.

Applications Solute concentration

1. You are working in a laboratory producing new antibiotics for
human and veterinary use. One compound with potential value
inhibits the action of prokaryotic ribosomes. The compound, how- 2. Most medically useful antibiotics interfere with either peptidogly-
ever, was shown to inhibit the growth of animal cells in culture. can synthesis or ribosome function. Why would the cytoplasmic
What is one possible explanation for its effect on animal cells? membrane be a poor target for antibacterial medications?
4 Dynamics of Prokaryotic Growth
KEY TERMS
Biofilm Polymer-encased
community of microorganisms.
Chemically Defined Medium
A culture medium composed of
Facultative Anaerobe Organism
that grows best if O2 is available, but
can also grow without it.
Generation Time The time it takes
exact quantities of pure chemicals; for a population to double in number.
generally used for specific
Obligate Aerobe Organism that
experiments when nutrients must be
requires molecular oxygen (O2).
precisely controlled.
Obligate Anaerobe Organism that
Complex Medium A culture
cannot multiply, and is often killed,
medium that contains protein
in the presence of O2.
digests, extracts, or other ingredients
that vary in their chemical Plate Count Method to measure
composition. the concentration of viable cells by
determining the number of colonies
Differential Medium A culture
that develop from a sample added to
medium with an ingredient that
an agar plate.
certain microorganisms change
in a recognizable way; used to Pure Culture A population
differentiate microbes based on their descended from a single cell.
metabolic traits. Selective Medium A culture
Exponential (Log) Phase Stage medium with an ingredient that
in the growth curve during which inhibits the growth of microbes other
cells divide at a constant rate; than the one being sought.
Bacterial colonies on an agar plate.
generation time is measured during
this period of active multiplication.

A Glimpse of History
The greatest contributor to methods of cultivating bacteria was Robert
Koch (1843–1910), a German physician who combined a medical prac-
tice with a productive research career for which he received a Nobel Prize planets, it may resemble these microbes. Each species, however,
in 1905. Koch was primarily interested in identifying disease-causing has a limited set of environmental conditions in which it can grow;
bacteria, but to do this, he needed simple methods to isolate and grow even then, it will grow only if specific nutrients are available.
these particular species. Koch recognized that a single bacterial cell could Some prokaryotes can grow at temperatures above the boiling
multiply on a solid medium in a limited area to form a distinct visible point of water but not at room temperature. Others can grow only
mass of descendants, so he experimented with growing bacteria on the within an animal host, and then only in specific areas of that host.
cut surfaces of potatoes. Some species would not grow, however, because Because of the medical significance of some microbes, as
the potatoes did not contain enough nutrients, so Koch experimented well as the nutritional and industrial use of microbial by-products,
with methods to solidify any liquid nutrient medium. He used gelatin scientists must be able to grow microorganisms in culture. This
initially, but there were two major drawbacks—it melts at the temperature
is why it is important to understand the basic principles involved
preferred by many medically important microbes and some bacteria can
in prokaryotic growth while recognizing that much information is
digest it. In 1882, Fannie Hess, the wife of an associate of Koch, sug-
gested using agar. This solidifying agent was used to harden jelly at the yet to be discovered.
time and proved to be the perfect answer.
Today, we take pure culture techniques for granted because of their
relative ease and simplicity. Their development, however, had a major 4.1 ■ Principles of
impact on microbiology. By 1900, the agents causing most of the major
bacterial diseases of humans had been isolated and characterized.
Prokaryotic Growth
Learning Outcome

P
rokaryotes can be found growing even in the harshest cli- 1. Describe binary fission and how it relates to generation time and
mates and most severe conditions. Environments that no exponential growth.
unprotected human could survive, such as the ocean depths,
volcanic vents, and the polar regions, have thriving prokaryotic Prokaryotes generally multiply by the process of binary fission
species. Indeed many scientists believe that if life exists on other (figure  4.1). After a cell has increased in size and doubled its

82
 Part I Life and Death of Microorganisms 83

components, it divides. One cell divides into two, those two divide
to become four, those four become eight, and so on. In other
words, the increase in cell numbers is exponential. Because it is
neither practical, nor particularly meaningful, to determine the
Cell gets longer and relative size of the cells in a given population, microbial growth is
DNA replicates.
defined as an increase in the number of cells in a population. The
time it takes for a population to double in number is the generation
time. This varies greatly from species to species, and is influenced
by the conditions in which the cells are grown.
The exponential multiplication of bacteria has important
DNA is moved into
each future daughter health consequences. For instance, a mere 10 cells of a food-borne
cell and cross wall forms. pathogen in a potato salad, sitting for 4 hours in the warm sun at
a picnic, may multiply to more than 40,000 cells. A simple equa-
tion expresses the relationship between the number of cells in a
population at a given time (Nt), the original number of cells in the
population (N0), and the number of divisions those cells have
undergone during that time (n). If any two values are known, the
Cell divides into two cells.
third can be easily calculated from the equation:
Nt = N0 × 2n
In this example, assume that 10 cells of the pathogen were
initially in the potato salad and that the organism has a generation
Cells separate. time of 20 minutes. The first step is to determine the number of
cell divisions that will occur in the 4-hour time frame. Because the
organism divides every 20 minutes (3 times per hour) we know
that in 4 hours it will divide 12 times. Once we know the original
number of cells and the number of divisions, we can solve for Nt:

Daughter cells 10 × 212 = Nt = 40,960

FIGURE 4.1 Binary Fission After 4 hours, our potato salad will have 40,960 cells of the patho-
? How does the process of binary fission relate to the generation gen (table  4.1). Keep this in mind, and your potato salad in a
time? cooler, the next time you go to a picnic!

TABLE 4.1 Example of Exponential Growth

Number of
Time in Minutes (t) Initial Population (N 0) Generations (n) 2n Population N0 × 2n

0 10 0 10
20 10 1 21 (= 2) 20
2
40 10 2 2 (= 2 × 2) 40
60 (1 hour) 10 3 23 (= 2 × 2 × 2) 80
4
80 10 4 2 (= 2 × 2 × 2 × 2) 160
100 10 5 25 (= 2 × 2 × 2 × 2 × 2) 320
6
120 (2 hours) 10 6 2 640
140 10 7 27 1,280
160 10 8 28 2,560
180 (3 hours) 10 9 29 5,120
10
200 10 10 2 10,240
220 10 11 211 20,480
12
240 (4 hours) 10 12 2 40,960
84 Chapter 4 Dynamics of Prokaryotic Growth

MicroByte
Escherichia coli has a generation time of 20 minutes in ideal
conditions; Mycobacterium tuberculosis needs at least 12 hours to
double.

MicroAssessment 4.1
Most prokaryotes multiply by binary fission. The time required for a
population to double in number is the generation time.
1. Explain why microbial growth refers to populations rather than
cell size.
2. If a bacterium has a generation time of 30 minutes, and you start
with 100 cells at time 0, how many cells will you have in 30, 60,
90, and 120 minutes? +

4.2 ■ Prokaryotic Growth

in Nature
FIGURE 4.2 Biofilm on a Stainless Steel Surface A biofilm
Learning Outcomes is a polymer-encased community of microorganisms.
2. Describe a biofilm, and give one positive and one negative impact ? Why would microbes in biofilms be more resistant to antibiotics
that biofilms have on humans. and disinfectants than their planktonic counterparts?
3. Explain why microbes that grow naturally in mixed communities
sometimes cannot be grown in pure culture.
are due to biofilms. In fact, the majority of bacterial infections
Historically, microorganisms have been studied by growing them seem to involve biofilms. Treatment of these infections is difficult
in the laboratory. Scientists now recognize, however, that the because microbes within the protective EPS often resist the effects
dynamic and complex conditions of the natural environment, of antibiotics as well as the body’s defenses. Biofilms are also
which differ greatly from the conditions in the laboratory, have important in industry, where their accumulations in pipes, drains,
profound effects on microbial growth and behavior. In fact, micro- and cooling water towers can interfere with processes as well as
bial cells are able to adjust to their surroundings by sensing vari- damage equipment. Again, the structure of the biofilm shields the
ous chemicals and then responding to that input by producing microbes growing within it, so the bacteria in a biofilm may be
materials appropriate for the situation. hundreds of times more resistant to disinfectants than are their
planktonic counterparts. disinfectants, p. 108
Although biofilms can be damaging, they also can be benefi-
Biofilms cial. Many bioremediation efforts, which use microbes to degrade
In nature, prokaryotes can live suspended in an aqueous environ- harmful chemicals, are enhanced by biofilms. So as some indus-
ment, but most attach to surfaces and live in polymer-encased tries are exploring ways to destroy biofilms, others, such as waste-
communities called biofilms (figure 4.2). Biofilms cause the slip- water treatment facilities, are looking for ways to foster their
periness of rocks in a stream bed, the slimy “gunk” that coats development. bioremediation, p. 744 wastewater treatment, p. 736
kitchen drains, the scum that gradually accumulates in toilet
bowls, and the dental plaque that forms on teeth. Biofilm forma-
Interactions of Mixed
tion begins when planktonic (free-floating) prokaryotic cells move
to a surface and adhere, where they then multiply and release Microbial Communities
polysaccharides, DNA, and other hydrophilic polymers to which Prokaryotes in the environment regularly grow in close associa-
unrelated cells may attach and grow (figure  4.3). The meshlike tions with many different species. Sometimes the interactions are
accumulation of these polymers, referred to as extracellular poly- cooperative, even fostering the growth of species that otherwise
meric substances (EPS), gives a biofilm its characteristic slimy could not survive. For example, organisms that cannot multiply in
appearance. dental plaque, pp. 63, 573 the presence of O2 will grow in the mouth if neighboring microbial
Surprisingly, biofilms are not random mixtures of microbes in cells consume that gas; one species creates a microenvironment in
a layer of EPS, but instead have characteristic formations with which the other can thrive. In a similar manner, the metabolic
channels through which nutrients and wastes pass. Cells commu- wastes of one organism can serve as nutrients for another. Often,
nicate with one another by synthesizing and responding to chemi- however, microbes in a community compete for nutrients, and
cal signals—an exchange important in establishing structure. some even resort to a type of biological warfare, synthesizing toxic
Biofilms are more than just an unsightly annoyance. Dental compounds that inhibit competitors. Understandably, the condi-
plaque leads to tooth decay and gum disease. Even troublesome, tions in these close associations are exceedingly difficult to repro-
persistent ear infections and the complications of cystic fibrosis duce in the laboratory. microbial competition and antagonism, p. 720
 Part I Life and Death of Microorganisms 85

Planktonic bacteria Bacteria multiply Other bacteria may Cells communicate Some cells detach
move to the surface and produce attach to the EPS and create channels and then move to
and adhere. extracellular polymeric and grow. in the EPS that allow other surfaces to
substances (EPS). nutrients and waste create additional
products to pass. biofilms.

FIGURE 4.3 Development of a Biofilm

? What are extracellular polymeric substances (EPS)?

activities of a particular species. But although the results are much

MicroAssessment 4.2
easier to interpret when studying pure cultures, the organisms
Biofilms have a characteristic architecture with channels through sometimes behave differently than they do in their natural envi-
which nutrients and wastes can pass. In nature, prokaryotes often
ronment, as discussed earlier. Another complicating issue is that
grow in associations with many different species.
only an estimated 1% of all prokaryotes can currently be grown in
3. Water bowls left out for pets sometimes develop a slimy layer if culture successfully. This makes it exceedingly difficult to study
not washed regularly. What causes the slime?
the vast majority of microorganisms. Fortunately for humanity,
4. Describe a situation in which the activities of one species benefit most known medically significant bacteria can be grown in pure
another.
culture.
5. Why would bacteria in a biofilm be more resistant to harmful To work with a pure culture, all containers, media, and instru-
chemicals? +
ments must be sterile, or free of microbes, prior to use. These are
then handled using aseptic techniques, procedures that minimize
the chance of other organisms being accidentally introduced. The
medium the cells are grown in, or on, is called a culture medium.
4.3 ■ Obtaining a Pure Culture It consists of nutrients dissolved in water, and can be a liquid broth
or a solid gel. aseptic techniques, p. 85 sterilization, p. 108
Learning Outcome
4. Describe how the streak-plate method is used to obtain a pure
culture, and how the resulting culture can be stored.
Growing Microorganisms
In the laboratory, prokaryotes are generally isolated and grown in on a Solid Medium
pure culture. A pure culture is a population descended from a The basic requirements for obtaining a pure culture are a solid
single cell and therefore separated from all other species. Work- culture medium, a container to hold and maintain the medium in
ing with pure cultures makes it easier to identify and study the an aseptic condition, and a method to separate individual microbial
86 Chapter 4 Dynamics of Prokaryotic Growth

very few microbes can degrade agar. It is not destroyed at high

temperatures and can therefore be sterilized by heating, a process
that also liquefies it. Melted agar will stay liquid until cooled to a
temperature below 45°C. Therefore, nutrients that would be
destroyed at high temperatures can be added at lower temperatures
before the agar hardens. Once solidified, an agar medium will
remain so until heated above 95°C. Thus, unlike gelatin—which is
liquid at 37°C—agar remains solid over the entire temperature
range at which most microbes grow. polysaccharide, p. 31
A solid culture medium is contained in a Petri dish—a
two-part, covered container made of glass or plastic. While not
airtight, the Petri dish does exclude airborne contaminants. A cul-
ture medium in a Petri dish is commonly referred to as a plate of
that medium—for example, a nutrient agar plate or, more simply,
an agar plate.

FIGURE 4.4 Colonies Growing on Agar Medium

? What is the purpose of agar in the medium?
The Streak-Plate Method
The streak-plate method is the simplest and most commonly used
technique for isolating prokaryotes (figure 4.5). A sterile inoculat-
ing loop is dipped into a microbe-containing sample and then lightly
cells. A single prokaryotic cell, supplied with the right nutrients drawn several times across the surface of an agar plate, creating a
and conditions, will multiply on the solid medium in a limited area set of parallel streaks covering approximately one-third of the agar.
to form a colony, a distinct mass of cells (figure  4.4). About The loop is then sterilized and a new series of parallel streaks is
1 million cells are required for a colony to be easily visible to the made across and at an angle to the previous ones, covering another
naked eye. surface section. This drags some of those cells streaked onto the
Agar, a polysaccharide extracted from marine algae, is used first portion over to a fresh section, effectively inoculating it with
to solidify culture media. Unlike gelatin and other gelling agents, a diluted sample. The loop is sterilized again, and another set of

1 Sterilize 4 Sterilize 6 Sterilize

loop. loop. loop.

2 Dip loop
into culture.

3 Streak first area. 5 Streak second area.

Starting point

Agar containing
nutrients
 Part I Life and Death of Microorganisms 87

parallel streaks is made, dragging into a third area some of the

organisms that had been moved into the second section. The goal is Stationary

Number of cells (logrithmic scale)

to reduce the number of cells being spread with each successive 1010 phase Death
series of streaks. By the third set of streaks, cells should be sepa- phase
rated enough so that distinct, well-isolated colonies will form. 108 Phase of
Log or prolonged decline
exponential
106 phase
Maintaining Stock Cultures
104
Once a pure culture has been obtained, it can be maintained as a
stock culture, a culture stored for use as an inoculum in later proce- Lag
102 phase
dures. Often, a stock culture is stored in the refrigerator as growth
on the surface of an agar slant (a tube of agar that was held at an 100
angle as it solidified). For long-term storage, stock cultures can be Time (hr) (days/months/years)
frozen at –70°C in a glycerol-containing solution that prevents ice
crystals from forming and damaging cells. Alternatively, cells can FIGURE 4.6 Growth Curve The growth curve is characterized by
be freeze-dried. five distinct stages: lag phase, exponential or log phase, stationary
phase, death phase, and phase of prolonged decline.

MicroAssessment 4.3 ? During which phase is generation time measured?

Only an estimated 1% of prokaryotes can be grown in culture. Agar

is used to solidify nutrient-containing broth. The streak-plate method
is used to obtain a pure culture. Stock cultures are stored in the
refrigerator or frozen. 4.4 ■ Prokaryotic Growth in
6. What properties of agar make it ideal for use in culture media? Laboratory Conditions
7. To identify the causative agent of a given illness, a pure culture
is often needed. How is a streak plate used to obtain a pure Learning Outcome
culture?
5. Describe the stages of a growth curve, and compare this closed
8. What might be a reason that pathogens can be grown in pure system to colony growth and continuous culture.
culture more often than environmental organisms? +
In the laboratory, bacteria and archaea are typically grown either
on agar plates or in tubes or flasks of broth. These are considered
closed systems, or batch cultures, because nutrients are not
renewed, nor are wastes removed. As the cells grow in this type of
system, the population increases in a distinct pattern of stages, and
then declines. This characteristic pattern is called a growth curve.
To maintain cells in a state of continuous growth, nutrients
must be continuously added and waste products removed. This is
an open system, or continuous culture.

Streak final area.

7
The Growth Curve
A growth curve is characterized by five distinct
stages—lag phase, exponential or log phase, stationary
phase, death phase, and phase of prolonged decline
(figure 4.6).
8 Isolated
colonies Lag Phase
develop
after incubation. When a dilute culture is transferred into a different medium,
the cell number does not immediately increase. During this lag
phase cells begin synthesizing enzymes required for growth.
The length of the lag phase depends on the conditions in the
FIGURE 4.5 The Streak-Plate Method The successive streaks original culture and the new medium. If cells are transferred into
dilute the cells. By the third set of streaks, cells should be sepa- a medium that contains fewer nutrients, the lag phase will be lon-
rated enough so that isolated colonies develop after incubation. ger because the cells must begin making amino acids or other
? What is the purpose of obtaining isolated colonies? components not supplied in the new medium. A similar situation
occurs when a stock culture is inoculated into fresh medium.
88 Chapter 4 Dynamics of Prokaryotic Growth

Exponential Phase (Log Phase) Death Phase

During the exponential or log phase, cells divide at a constant The death phase is the period when the total number of viable cells
rate. This is when the generation time is measured. in the population decreases as cells die off at a constant rate. Like
The exponential phase is medically important because bacte- cell growth, death is exponential, but the rate is usually much slower.
ria are most sensitive to antimicrobial medications at this stage.
Many of these medications target processes primarily active when Phase of Prolonged Decline
bacteria are multiplying. In many cases, a fraction of the cell population survives the death
From a commercial standpoint, the exponential phase is phase. These cells have adapted to tolerate the worsened condi-
important because some molecules made by growing cells are tions and are able to multiply for at least a short time, using the
valuable. For instance, amino acids can be sold as nutritional nutrients released from the dead cells. As the conditions continue
supplements, and microbial waste products such as ethanol are to deteriorate during this phase of prolonged decline, most of
used as biofuels. The small molecules made by cells as they mul- these survivors then die. However, the few progeny better
tiply are called primary metabolites. equipped for survival can grow. This dynamic process generates
In the later stages of exponential growth, nutrients gradually successive waves of slightly modified populations, each more fit
become depleted and waste products accumulate. Cells’ activities to survive than the previous ones. Thus, the statement “survival of
shift as this occurs. If the cells are able to form endospores, they the fittest” holds true even for closed cultures of microbes.
initiate the process of sporulation. If they cannot, they still alter
their activities to prepare for starvation conditions. Compounds
that begin accumulating at this stage are made for purposes other Colony Growth
than growth and are called secondary metabolites (figure 4.7). Growth of a microbial colony on a solid medium involves many of
Commercially, the most valuable of these are antibiotics. the same features as growth in liquid, but it is marked by some
important differences. After a lag phase, cells multiply exponentially
Stationary Phase and eventually compete with one another for available nutrients.
Unlike the situation in a liquid culture, however, the position of a
Cells enter the stationary phase when the nutrient levels are too
single cell within a colony determines its environment. Cells multi-
low to sustain growth. The total number of viable cells in the
plying on the edge of the colony face relatively little competition for
population remains relatively constant, but some cells are dying
O2 and nutrients. In the center of the colony, meanwhile, the high
while others are multiplying. How can cells multiply when they density of cells rapidly depletes available O2 and nutrients. Wastes
have exhausted their supply of nutrients? Dead cells often burst, such as acids accumulate, and these can be toxic. As a consequence,
releasing nutrients that then fuel the growth of other cells. cells at the edge of the colony may be growing exponentially, while
During the stationary phase, the viable cells continue to syn- those in the center may be in the death phase. Cells in locations
thesize secondary metabolites and maintain the altered properties between these two extremes may be in the stationary phase.
they demonstrated in late log phase. The length of time cells
remain in the stationary phase varies, depending on the species
and environmental conditions. Some populations remain in the Continuous Culture
stationary phase for only a few hours, whereas others remain for Microbial cells can be kept in a state of continuous growth by
days or longer. using a chemostat. This device continually drips fresh medium
into a broth culture contained in a chamber. With each drop that
enters, an equivalent volume—containing cells, wastes, and spent
medium—leaves through an outlet. By manipulating the nutrient
Number of cells (logarithmic scale)

Stationary content of the medium and the speed at which it enters the cham-
ber, a constant growth rate and cell density can be maintained.
Synthesis of metabolites

This makes it possible to study a uniform population’s response to

different nutrient concentrations or environmental conditions.
Log

MicroAssessment 4.4
Primary
metabolite Secondary When grown in a closed system, a prokaryotic population goes
Lag metabolite through five distinct phases: lag, exponential or log, stationary, death,
and prolonged decline. Cells within a colony may be in any one of the
phases, depending on their relative location. Chemostats are used to
Time (hr) study uniform populations of growing cells.
9. Explain the difference between the lag and exponential phases.
FIGURE 4.7 Primary and Secondary Metabolite Production 10. Describe how a chemostat keeps cells in a state of continuous
Primary metabolites are synthesized during the period of active growth.
multiplication. Secondary metabolites begin to be synthesized in late
log phase. 11. Why would a compound that prevents bacteria from growing
interfere with the function of the antibiotic penicillin? +
? What is the most commercially valuable secondary metabolite?
 Part I Life and Death of Microorganisms 89

■
4.5 ■ Environmental Factors That Psychrophiles have an optimum between –5°C and 15°C.
These organisms grow in the cold Arctic and Antarctic
Influence Microbial Growth regions and in lakes fed by glaciers.
■ Psychrotrophs have an optimum between 20°C and 30°C,
Learning Outcomes
but grow well at lower temperatures. They are an important
6. Describe the importance of a prokaryote’s requirements for
temperature, O2, pH, and water availability, and define the terms cause of spoilage in refrigerated foods. food spoilage, p. 756
that express these requirements. ■ Mesophiles have an optimum between 25°C and about 45°C.
7. Explain the significance of reactive oxygen species, and describe E. coli and most other common bacteria are in this group.
the mechanisms cells use to protect against their effects.

Environmental Factors That

As a group, prokaryotes inhabit nearly every environment on TABLE 4.2
Influence Microbial Growth
earth. Microbes we associate with disease and rapid food spoilage
live in habitats that humans consider quite comfortable. Some, Environmental Factor/
however, live in harsh environments that would kill most other Descriptive Terms Characteristics
organisms. Most of the examples in this latter group, called Temperature Thermostability appears to be
extremophiles (phile means “loving”), are in the domain Archaea. due to protein structure.
Recognizing the major environmental factors that influence Psychrophile Optimum temperature between
microbial growth—temperature, atmosphere, pH, and water –5°C and 15°C.
availability—is essential for studying microbes and helps us Psychrotroph Optimum temperature between
understand their roles in the complex ecology of the planet. These 20°C and 30°C, but grows well at
factors and their associated characteristics are summarized in refrigeration temperatures.
table 4.2. Mesophile Optimum temperature between
25°C and 45°C.
Thermophile Optimum temperature between
Temperature Requirements 45°C and 70°C.

Each microbial species has a well-defined temperature range in Hyperthermophile Optimum temperature of 70°C or
greater.
which it grows. Within this range is the optimum growth
temperature, the temperature at which the organism multiplies Oxygen (O2) Availability Oxygen (O2) requirement/
most rapidly. As a general rule, this optimum is close to the upper tolerance reflects the organism’s
energy-harvesting mechanisms
limit of the organism’s temperature range. and its ability to inactivate
Prokaryotes are commonly divided into five groups based on reactive oxygen species.
their optimum growth temperatures (figure 4.8). Note, however, Obligate aerobe Requires O2.
that this merely represents a convenient organization scheme. In Facultative anaerobe Grows best if O2 is present, but
reality, no sharp dividing line exists between each group. can also grow without it.
Furthermore, not every organism in a group can grow in the entire Obligate anaerobe Cannot grow in the presence of O2.
temperature range typical for its group. Microaerophile Requires small amounts of O2,
but higher concentrations are
inhibitory.
Aerotolerant anaerobe Indifferent to O2.
Hyperthermophile (obligate fermenter)
Mesophile Thermophile
pH Prokaryotes that live in pH
extremes maintain a near-neutral
Growth rate

Psychrotroph internal pH by pumping protons

Psychrophile out of or into the cell.
Neutrophile Multiplies in the range of pH 5 to 8.
Acidophile Grows optimally at a pH below 5.5.
Alkalophile Grows optimally at a pH above 8.5.
Water Availability Prokaryotes that can grow in
high-solute solutions maintain
–10 0 10 20 30 40 50 60 70 80 90 100 110 120
the availability of water in the
Temperature (°C) cell by increasing their internal
solute concentration.
FIGURE 4.8 Temperature Requirements for Growth Halotolerant Can grow in relatively high-salt
Prokaryotes are commonly divided into five groups based on their solutions, up to approximately
optimum growth temperatures. This graph depicts a typical example 10% NaCl.
of each group. Note that the optimum temperature, the point at
which the growth rate is highest, is near the upper limit of the range. Halophile Requires high levels of sodium
chloride.
? Most pathogens fall into which group on this chart?
90 Chapter 4 Dynamics of Prokaryotic Growth

Pathogens, adapted to growth in the human body, typically diffusion, the level of O2 in the tube is high at the top, whereas the
have an optimum between 35°C and 40°C. Mesophiles that bottom portion is anaerobic. The cells grow in the region that has
inhabit soil, a colder environment, generally have a lower a suitable O2 level (table 4.3).
optimum, close to 30°C. Based on their O2 requirements, prokaryotes can be separated
■ Thermophiles have an optimum between 45°C and 70°C. into these groups:
These organisms commonly live in hot springs and compost ■ Obligate aerobes have an absolute requirement for oxygen
heaps. composting, p. 743 (O2). They use it in aerobic respiration, an energy-harvesting
■ Hyperthermophiles have an optimum of 70°C or greater. These process. This and other ATP-generating pathways will be
are usually members of the Archaea. One member, isolated from discussed in chapter 6. An example of an obligate aerobe is
the wall of a hydrothermal vent deep in the ocean, has a maxi- Micrococcus luteus, which is common in the environment.
mum growth temperature of 121°C, the highest yet recorded! aerobic respiration, pp. 134, 144 ATP, pp. 24, 128

Why can some prokaryotes withstand very high temperatures ■ Facultative anaerobes grow better if O2 is present, but can
but most cannot? As a general rule, proteins from thermophiles are also grow without it. The term “facultative” means that the
not denatured at high temperatures. This thermostability is due to organism is flexible, in this case in its requirements for O2.
the amino acid sequence of the protein. This controls the number Facultative anaerobes use aerobic respiration if O2 is avail-
and position of the bonds that form within the protein, which in able, but resort to alternative types of metabolism if it is not.
turn determines its three-dimensional structure. Heat-stable Growth is faster when O2 is present because aerobic respira-
enzymes that degrade fats and other proteins are used in high- tion yields the most ATP. E. coli is one of the most common
temperature detergents. protein denaturation, p. 29 facultative anaerobes in the large intestine. fermentation,
pp. 134, 147 anaerobic respiration, pp. 134, 144

Temperature and Food Preservation ■ Obligate anaerobes cannot multiply if O2 is present; in fact,
Refrigeration temperatures (approximately 4°C) slow spoilage they are often killed by even brief exposure to air. Obligate
because they limit the multiplication of otherwise fast-growing anaerobes harvest energy using processes other than aerobic
mesophiles. Psychrophiles and psychrotrophs can still grow, how- respiration; the details of these will be covered in chapter 6. Most
ever, so refrigerated foods will still spoil, but it happens more inhabitants of the large intestine are obligate anaerobes, as is the
slowly. low-temperature storage, p. 121 food spoilage, p. 756 bacterium that causes botulism—Clostridium botulinum.
Foods and other perishable products that withstand below- ■ Microaerophiles require small amounts of O2 (2% to 10%)
freezing temperatures can be frozen for long-term storage. It is for aerobic respiration; higher concentrations are inhibitory.
important to recognize, however, that freezing is not an effective An example is Helicobacter pylori, which causes gastric and
means of destroying microbes. Recall that freezing is routinely duodenal ulcers.
used to preserve stock cultures. ■ Aerotolerant anaerobes are indifferent to O2. They can
grow in its presence, but do not use it to harvest energy. They
Temperature and Disease are also called obligate fermenters, because fermentation is
The temperatures of different parts of the human body vary signifi- their only metabolic option. An example is Streptococcus
cantly. For example, the heart, brain, and gastrointestinal tract are near pyogenes, which causes strep throat.
37°C, but the temperature of the extremities is lower. For these rea-
sons, some microbes cause disease more readily in certain body parts. MicroByte
Hansen’s disease (leprosy) typically involves the coolest regions (ears, Over half of all the cytoplasm on earth is probably in anaerobic
hands, feet, and fingers) because the causative bacterium, microbes!
Mycobacterium leprae, grows best at these lower temperatures. The
same situation applies to syphilis, in which lesions appear on the Reactive Oxygen Species (ROS)
genitalia and then on the lips, tongue, and throat. Indeed, for more than
When organisms use O2 in aerobic respiration, harmful derivatives
30 years the major treatment of syphilis was to induce fever by delib-
called reactive oxygen species (ROS) form as by-products.
erately introducing the agent that causes malaria, which results in very
Examples of these molecules include superoxide (O2–) and hydro-
high fevers. Hansen’s disease, p. 650 syphilis, p. 626 fever, p. 351
gen peroxide (H2O2). Reactive oxygen species can damage cell
components, so cells that grow aerobically must have mechanisms
Oxygen (O2) Requirements to protect against them. Obligate anaerobes typically do not have
Like humans, some prokaryotes have an absolute requirement for these mechanisms, but there are exceptions.
O2. Others thrive in environments that are anaerobic, meaning Virtually all organisms that grow in the presence of O2 pro-
little or no O2 is present. One way of determining an organism’s duce the enzyme superoxide dismutase, which inactivates super-
O2 requirement is to grow it in a shake tube. To do this, a tube of oxide by converting it to O2 and hydrogen peroxide. Nearly all
nutrient agar is boiled, which both melts the agar and drives off these organisms produce the enzyme catalase as well, which con-
the O2. The agar is then allowed to cool to just above its solidify- verts hydrogen peroxide into O2 and water. An important exception
ing temperature. Next, the test organism is added and dispersed by is the aerotolerant anaerobes. The fact that they do not produce
gentle shaking or swirling. The agar medium is allowed to harden catalase is useful in the laboratory. A simple test for the enzyme
and the tube is incubated. Because the solidified agar slows gas can be used to distinguish two groups of medically important
 Part I Life and Death of Microorganisms 91

TABLE 4.3 Oxygen (O2) Requirements of Prokaryotes

Obligate Facultative Obligate Microaerophile Aerotolerant
aerobe anaerobe anaerobe anaerobe

Bacterial
growth

Growth Grows only when O2 is Grows best when O2 Cannot grow when O2 Grows only if small Grows equally well
characteristics available. is available, but also is present. amounts of O2 are with or without O2.
grows without it. available.
Use of O2 Requires O2 for Uses O2 for respiration, Does not use O2. Requires O2 for Does not use O2.
in energy- respiration. if available. respiration.
harvesting
processes
Typical Produces superoxide Produces superoxide Does not produce Produces some Produces superoxide
mechanisms to dismutase and catalase. dismutase and catalase. superoxide dismutase superoxide dismutase but not
protect against or catalase. dismutase and catalase.
reactive oxygen catalase.
species

Gram-positive cocci that grow aerobically—Staphylococcus spe- Water Availability

cies, which are catalase positive, and Streptococcus species, which
All microorganisms require water for growth. Even if water is
are catalase negative. catalase test, p. 243
present, however, it may not be available in certain environments.
Dissolved substances such as salt (NaCl) and sugars, for example,
pH interact with water molecules, making them unavailable to the
cell. In many environments, particularly in certain natural habitats
Each prokaryotic species can survive within a range of pH values,
such as salt marshes, prokaryotes are faced with this situation. If
and within this range is its pH optimum. Despite the pH of the
the solute concentration is higher in the medium than in the cell,
external environment, however, cells maintain a constant internal
water diffuses out of the cell due to osmosis. This causes the cyto-
pH, typically near neutral. Many that grow in acidic environments
plasm to dehydrate and shrink from the cell wall, a phenomenon
quickly pump out protons that enter the cell, whereas those that
called plasmolysis (figure 4.9). solute p. 54 osmosis, p. 54
grow in alkaline conditions bring in protons. pH, p. 23
Most microbes are neutrophiles—they live and multiply
within the range of pH 5 (acidic) to pH 8 (basic), and have a pH
Dissolved substances Cytoplasmic membrane
optimum near neutral (pH 7). Food preservation methods such as (solute) pulls away from the
pickling inhibit bacterial growth by increasing the acidity of the cell wall (plasmolysis).
food.
While most neutrophiles cannot withstand highly acidic con- Cytoplasmic
membrane
ditions, one medically important bacterium has found a way.
Helicobacter pylori grows in the stomach, sometimes causing
ulcers. It decreases the acidity of its immediate surroundings by
producing urease, an enzyme that splits urea into carbon dioxide
and ammonia. The ammonia neutralizes any stomach acid sur- Cell wall
rounding the cell. ulcers, p. 580
Acidophiles grow optimally at a pH below 5.5. Picrophilus Water flows
out of cell
oshimae, a member of the Archaea, has an optimum pH of less
than 1! This prokaryote, which was isolated from the dry, acid FIGURE 4.9 Effects of Solute Concentration on Cells
soils of a gas-emitting volcanic fissure in Japan, has an unusual The cytoplasmic membrane allows water molecules to pass through
cytoplasmic membrane that is unstable at a pH above 4.0. freely. If the solute concentration is higher outside of the cell, water
Alkaliphiles grow optimally at a pH above 8.5. They often moves out.
live in alkaline lakes and soils. ? What is plasmolysis?
92 Chapter 4 Dynamics of Prokaryotic Growth

The growth-inhibiting effect of high salt and sugar concentra-

Representative Functions
tions is used in food preservation. High levels of salt are added to TABLE 4.4
of the Major Elements
preserve such foods as bacon, salt pork, and anchovies. Many
foods with a high sugar content, such as jams, jellies, and honey, Chemical Function
are naturally preserved. food preservation, p. 759
Carbon, oxygen, and Component of amino acids, lipids,
Although many microbes are inhibited by high concentrations
hydrogen nucleic acids, and sugars
of salt, some withstand or even require it. Microbes that tolerate
Nitrogen Component of amino acids and nucleic
high concentrations of salt, up to approximately 10% NaCl, are
acids
halotolerant (halo means “salt”). Staphylococcus species, which
Sulfur Component of some amino acids
reside on the dry salty environment of the skin, are an example.
Halophiles require high levels of sodium chloride. Many marine Phosphorus Component of nucleic acids, membrane
lipids, and ATP
bacteria are mildly halophilic, requiring concentrations of approx-
imately 3% sodium chloride. Certain members of the Archaea are Potassium, Required for the functioning of certain
magnesium, and enzymes; additional functions as well
extreme halophiles, requiring 9% sodium chloride or more.
calcium
Extreme halophiles are found in environments such as the salt flats
Iron Part of certain enzymes
of Utah and the Dead Sea.

MicroAssessment 4.5
A prokaryotic species can be categorized according to its optimum the essential components of proteins, carbohydrates, lipids, and
growth temperature. A species can also be grouped according to its O2
nucleic acids (table 4.4). elements, p. 17
requirements. Most microbes grow best at a near-neutral pH, although
some prefer acidic conditions, and others grow best in alkaline The source of carbon distinguishes different groups of pro-
conditions. Halophiles require high-salt conditions. karyotes. Heterotrophs use organic carbon (hetero means “differ-
12. Clostridium paradoxum grows optimally at 55oC, pH 9.3; it
ent” and troph means “nourishment”). Medically important
will not grow in the presence of O2. How should this bacterium bacteria are typically heterotrophs. Autotrophs (auto means
be categorized with respect to its temperature, pH, and O2 “self”) use inorganic carbon in the form of carbon dioxide (CO2).
requirements? They play a critical role in the cycling of carbon in the environ-
13. What is the function of the enzyme catalase? ment because they can convert inorganic carbon to an organic
14. Why would hydrogen peroxide be an effective disinfectant? + form, the process of carbon fixation. Without carbon fixation, the
earth would quickly run out of organic carbon, which is essential
to life. carbon cycle, p. 725
Nitrogen is needed to make amino acids and nucleic acids.
4.6 ■ Nutritional Factors That Some prokaryotes use nitrogen gas (N2) as a nitrogen source, con-
Influence Microbial Growth verting it to ammonia and then incorporating that into cellular
material. This process, nitrogen fixation, is unique to prokary-
Learning Outcomes otes. Like carbon fixation, it is essential to life because once the
8. List the required elements and examples of common sources. nitrogen is incorporated into cellular material such as amino acids,
9. Explain the significance of a limiting nutrient. other organisms can easily use it. Many microbes use ammonia as
10. Explain why fastidious microbes require growth factors. a nitrogen source. Some convert nitrate to ammonia, which is then
11. Describe the energy and carbon sources used by photo- incorporated into cellular material. All of these nitrogen uses are
autotrophs, chemolithoautotrophs, photoheterotrophs, and important steps in the nitrogen cycle nitrogen cycle, p. 726
chemoorganoheterotrophs. Sulfur is a component of some amino acids. Many microbes
use inorganic sulfur sources such as sulfate, but others require
Growth of any prokaryote depends not only on a suitable physical organic sources such as sulfur-containing amino acids.
environment, but also on the availability of nutrients. The cells Phosphorus is a component of nucleic acids, membrane
use the nutrients to synthesize the components discussed in lipids, and ATP. As with sulfur, many organisms can use inor-
chapter 3—including lipid membranes, cell walls, proteins, and ganic phosphorus sources such as phosphate. Some, however,
nucleic acids. These are made from subunits such as phospholipids, require organic sources, such as phosphorus-containing cell
sugars, amino acids, and nucleotides. In turn, each of these subunits is components.
composed of a variety of chemical elements, including carbon and Other elements, including potassium, magnesium, calcium,
nitrogen. What sets the prokaryotic world apart from all other forms of and iron, are required for some enzyme functions. A variety of
life is their remarkable ability to use diverse sources of these elements. inorganic and organic sources may be used by microbes.
Phosphorus and iron are important ecologically because
they are often limiting nutrients—meaning they are available at
Required Elements the lowest concentration relative to need. To understand this
Chemical elements that make up cell constituents are called major concept, think about using a recipe to make chocolate chip cook-
elements; these include carbon, oxygen, hydrogen, nitrogen, sulfur, ies. Just as the quantity of chocolate chips available would deter-
phosphorus, potassium, magnesium, calcium, and iron. They are mine the number of batches you could make (assuming the other
 Part I Life and Death of Microorganisms 93

ingredients are on hand), a limiting nutrient dictates the maxi- Nutritional Diversity
mum level of microbial growth. This concept is important eco-
Microbiologists often group prokaryotes according to the energy
logically, as illustrated when algal blooms in a small Seattle lake
and carbon sources they utilize (table 4.5):
resulted in a murky mess. To solve the problem, a chemical
treatment was used to remove excess phosphate in the lake. By ■ Photoautotrophs use the energy of sunlight along with CO2
decreasing the level of phosphorus—the limiting nutrient—algal in the atmosphere to make organic compounds. They are
growth was restricted. primary producers, meaning they support other forms of life
Trace elements are required in such small amounts that most by fixing carbon. Cyanobacteria are important primary pro-
natural environments, including water, have sufficient levels to ducers that inhabit soil and aquatic environments. Many fix
support microbial growth. Trace elements include cobalt, zinc, nitrogen as well, providing another indispensable role in the
copper, molybdenum, and manganese. biosphere. primary producers, p. 719
■ Photoheterotrophs use the energy of sunlight and derive
Growth Factors their carbon from organic compounds. Some are facultative
Some microbes cannot synthesize certain organic molecules such in their nutritional capabilities. For example, some mem-
as amino acids, vitamins, purines, or pyrimidines. Consequently, bers of a group called the purple nonsulfur bacteria grow
these organisms grow only if the molecule they cannot produce is anaerobically using light as an energy source and organic
available in the surrounding environment; the molecule is a compounds as a carbon source (photoheterotrophs). They
growth factor. purines, p. 32 pyrimidines, p. 32 can also grow aerobically in the dark using organic sources
A microbe’s growth factor requirements reflect its biosyn- of carbon and energy (chemoheterotrophs). purple nonsulfur
thetic capabilities. Most E. coli strains, for example, can use glu- bacteria, p. 260
cose as the raw material to synthesize all of their cell components, ■ Chemolithoautotrophs (lith means “stone”), often referred
so they do not need any growth factors. They multiply in a to simply as chemoautotrophs or chemolithotrophs, use inor-
medium containing only glucose and six different inorganic salts. ganic compounds for energy and derive their carbon from
In contrast, Neisseria species are less resourceful metabolically, CO2. These prokaryotes live in seemingly inhospitable places
and require numerous growth factors including vitamins and such as sulfur hot springs, which are rich in hydrogen sulfide,
amino acids. Bacteria such as Neisseria are fastidious, meaning and other environments that have reduced inorganic com-
they have complicated nutritional requirements. pounds (see Perspective 4.1). In some regions of the ocean
Fastidious bacteria are used to measure the quantity of vita- depths, near hydrothermal vents, chemoautotrophs serve
mins in food products. To do this, a well-characterized species that as essential primary producers, supporting rich communi-
requires a specific vitamin is inoculated into a medium that lacks ties in these habitats that lack sunlight (see figure  29.11).
the vitamin but is supplemented with a measured amount of the hydrothermal vents, p. 729
food product. The extent of growth of the bacterium is related to
■ Chemoorganoheterotrophs, also referred to as chemohet-
quantity of the vitamin in the product.
erotrophs or chemoorganotrophs, use organic compounds
for both energy and carbon. They are by far the most com-
Energy Sources mon group of microorganism associated with humans and
Organisms harvest energy either from sunlight or chemical com- other animals. Individual species of chemoheterotrophs
pounds, using processes discussed in chapter 6. Phototrophs differ in the number of organic compounds they can use.
obtain energy from sunlight (photo means “light”). They include For example, certain members of the genus Pseudomonas
plants, algae, and photosynthetic bacteria. Chemotrophs extract can derive carbon and/or energy from more than 80 differ-
energy from chemical compounds (chemo means “chemical”). ent organic compounds, including such unusual compounds
Mammalian cells, fungi, and many types of prokaryotes use as naphthalene (the ingredient associated with the smell
organic chemicals such as sugars, amino acids, and fatty acids as of mothballs). At the other extreme, some organisms can
energy sources. Some prokaryotes extract energy from inorganic degrade only a few compounds. Bacillus fastidiosus can use
chemicals such as hydrogen sulfide and hydrogen gas, an ability only urea and certain of its derivatives as a source of both
that distinguishes them from eukaryotes. carbon and energy.

TABLE 4.5 Energy and Carbon Sources Used by Different Groups of Prokaryotes
Type Energy Source Carbon Source

Photoautotroph Sunlight CO2

Photoheterotroph Sunlight Organic compounds
– 2+
Chemolithoautotroph Inorganic chemicals (H2, NH3, NO2 , Fe , H2S) CO2
Chemoorganoheterotroph Organic compounds (sugars, amino acids, etc.) Organic compounds
94 Chapter 4 Dynamics of Prokaryotic Growth

PERSPECTIVE 4.1
Can Prokaryotes Live on Only Rocks and Water?
Prokaryotes have been isolated from diverse pressures of 160 atmospheres (at sea level, the What do these organisms use for food? They
environments that previously were thought to pressure is 1 atmosphere). The discovery of apparently get their energy from the H2 pro-
be incapable of sustaining life. For example, these microbes suggests that thermophiles may duced in a reaction between groundwater and
members of the Archaea have been isolated be widespread in the earth’s crust. the iron-rich minerals in the rock. The ground-
from environments 10 times more acidic than Perhaps the most unusual environment water also contains dissolved CO2, which the
that of lemon juice. Other Archaea have been from which prokaryotes have been isolated is organisms use as a source of carbon. These
isolated from oil wells a mile below the sur- volcanic rock 1 mile below the earth’s surface prokaryotes apparently exist on nothing more
face of the earth at temperatures of 70°C and near the Columbia River in Washington State. than rocks and water!

mixture of amino acids and short peptides produced by digesting any

MicroAssessment 4.6
of a variety of different proteins. Other substances, such as extracts
Organisms require a source of major and trace elements. Heterotrophs use (the water-soluble components of a substance such as lean beef), are
an organic carbon source, and autotrophs use CO2. Phototrophs harvest
often added to provide vitamins and minerals. A common complex
energy from sunlight, and chemotrophs extract energy from chemicals.
medium—nutrient broth—consists of peptone and beef extract in
15. To prevent excess phosphate from entering lakes and streams, distilled water. If agar is added, then nutrient agar results.
certain laws govern the amount of phosphorus allowed in
Many medically important bacteria are fastidious, requiring a
laundry and dishwasher detergents. What can happen if
phosphorus levels in a lake increase? medium even richer than nutrient agar. Because of this, clinical
laboratories often use blood agar. As the name implies, this con-
16. How would your cells be categorized with respect to their carbon
and energy sources? tains red blood cells, a source of a variety of nutrients including
hemin; the medium contains other ingredients as well. A medium
17. Why would human-made materials (such as many plastics) be
degraded only slowly or not at all? + used for even more fastidious bacteria is chocolate agar, named
for its brownish appearance rather than its ingredients. Chocolate
agar contains lysed red blood cells and additional nutrients.

4.7 ■ Cultivating Prokaryotes Characteristics of Representative

TABLE 4.6
in the Laboratory Media Used to Cultivate Bacteria
Medium Characteristic
Learning Outcomes
12. Compare and contrast complex, chemically defined, selective, and Blood agar Complex medium used routinely in clinical
differential media. labs. Differential because colonies of hemolytic
13. Explain how aerobic, microaerophilic, and anaerobic conditions organisms are surrounded by a zone of red
can be provided. blood cell clearing. Not selective.

14. Describe the purpose of an enrichment culture. Chocolate agar Complex medium used to culture fastidious
bacteria, particularly those found in clinical
specimens. Not selective or differential.
By knowing the environmental and nutritional factors that influ-
Glucose-salts Chemically defined medium. Used in
ence the growth of specific prokaryotes, it is often possible to laboratory experiments to study nutritional
provide appropriate conditions for their cultivation. These include requirements of bacteria. Not selective or
a suitable growth medium and the proper atmosphere. differential.
MacConkey Complex medium used to isolate Gram-
General Categories of Culture Media agar negative rods that typically reside in the
Considering the diversity of prokaryotes, it should not be surpris- intestine. Selective because bile salts and
dyes inhibit Gram-positive organisms and
ing that hundreds of different types of media are available. Even
Gram-negative cocci. Differential because
so, some medically important organisms and most environmental the pH indicator turns pink-red when the
ones have not yet been grown on in the laboratory. Table 4.6 sum- sugar in the medium, lactose, is fermented.
marizes the characteristics of representative examples of media. Nutrient agar Complex medium used for routine
laboratory work. Supports the growth of
Complex Media a variety of nonfastidious bacteria. Not
A complex medium contains a variety of ingredients such as meat selective or differential.
juices and digested proteins, forming what might be viewed as a tasty Thayer-Martin Complex medium used to isolate Neisseria
soup for microbes. This type of medium is easy to make and is used species, which are fastidious. Selective
because it contains antibiotics that inhibit
for routine purposes. Although a specific amount of each ingredient
most organisms except Neisseria species. Not
is in the medium, the exact chemical compositions of those sub- differential.
stances can be highly variable. One common ingredient is peptone, a
 Part I Life and Death of Microorganisms 95

a problem in complex media because the amino acids and other

Ingredients in Two Types of Media
TABLE 4.7 natural components provide at least some buffering action.
That Support the Growth of E. coli
buffer, p. 23
Nutrient Broth Glucose-Salts Broth
(complex medium) (chemically defined medium)
Special Types of Culture Media
Peptone Glucose
To detect or isolate a species from a mixed population, it is often
Meat extract Dipotassium phosphate necessary to make that species more prevalent or obvious. For
Water Monopotassium phosphate these purposes selective and differential media are used. They can
Magnesium sulfate be either complex or chemically defined, depending on the needs
Ammonium sulfate of the microbiologist.
Calcium chloride
Selective Media
Iron sulfate
Selective media inhibit the growth of certain species, making it
Water
easier to isolate the one being sought. For example, Thayer-Martin
agar is used to isolate Neisseria gonorrhoeae from clinical speci-
mens. Thayer-Martin is chocolate agar to which three or more
Chemically Defined Media antimicrobial drugs have been added. The antimicrobials inhibit
Chemically defined media are composed of exact amounts of fungi, Gram-positive bacteria, and Gram-negative rods, but not
pure chemicals. This type of medium is generally used only for most N. gonorrhoeae strains. Because of this, the pathogen can
specific research experiments when the type and quantity of nutri- grow on the medium with little competition.
ents must be precisely controlled. A chemically defined medium MacConkey agar is used to isolate Gram-negative rods from
called glucose-salts supports the growth of E. coli, and contains various clinical specimens such as urine. In addition to containing
only those chemicals listed in table  4.7. The cells grow more peptones and other nutrients, this medium includes two inhibitory
slowly in this medium than in nutrient broth because they must compounds—crystal violet (a dye that inhibits Gram-positive bac-
synthesize all of their cell components from glucose. A more teria) and bile salts (a compound that inhibits most non-intestinal
elaborate recipe containing as many as 46 different ingredients bacteria).
must be used to make a chemically defined medium that supports
the growth of the fastidious bacterium Neisseria gonorrhoeae (the Differential Media
cause of gonorrhea). Differential media contain substances that certain microbes
To maintain the pH near neutral, buffers are often added to change in a recognizable way. Blood agar is differential because
culture media. They are especially important in defined media bacteria that produce a hemolysin—a protein that causes red
because some bacteria produce so much acid as a by-product of blood cells to burst—are surrounded by a zone of clearing called
metabolism that they inhibit their own growth. This is usually not hemolysis (figure 4.10). This readily observable characteristic is

Colony Zone of clearing

(a) (b)
FIGURE 4.10 Blood Agar This complex medium is differential for hemolysis. (a) A zone of complete clearing around a colony growing on
blood agar is called beta hemolysis. (b) A zone of greenish clearing is called alpha hemolysis.
? Which type of hemolysis characterizes Streptococcus pyogenes, the bacterium that causes strep throat?
96 Chapter 4 Dynamics of Prokaryotic Growth

have additional CO2. Some are even capnophiles, meaning they

require increased CO2. One of the simplest ways to build up the
level of CO2 in the incubation atmosphere is to use a candle jar.
For this system, the inoculated plates or tubes are added to the jar
and then, just before the container is closed, a candle within it is
lit. As the candle burns, it consumes some of the O2 in the air,
generating CO2 and H2O; the flame soon extinguishes because of
insufficient O2. However, about 17% O2 remains, enough to sup-
port the growth of obligate aerobes and prevent the growth of
obligate anaerobes. Special incubators are also available that
maintain CO2 at prescribed levels.

Microaerophilic
Microaerophilic prokaryotes typically require O2 concentrations
FIGURE 4.11 MacConkey Agar This complex medium is differen- less than what is achieved in a candle jar. These microbes are
tial for lactose fermentation and selective for Gram-negative rods. often incubated in a gastight container with a special disposable
? What specifically causes colonies of lactose fermenters to be dark packet; the packet holds chemicals that react with O2, reducing its
pink on MacConkey agar? concentration to approximately 5–15%.

Anaerobic
Obligate anaerobes are challenging to grow because of their
important medically because some pathogens can be distin- sensitivity to O2-containing environments. Those that can toler-
guished by their type of hemolysis. For example, the colonies of ate brief exposures to O2 are incubated in an anaerobe container
Streptococcus pyogenes (the cause of strep throat) produce a (figure 4.12). This is the same type of container used to incubate
clear zone of hemolysis called beta hemolysis. This makes them microaerophiles, but the chemical composition of the disposable
stand out from Streptococcus species that reside harmlessly in packet produces an anaerobic environment. An alternative
the throat. Many of these show alpha hemolysis, meaning their method is to use a semisolid culture medium that contains a
colonies are surrounded by a zone of greenish partial clearing;
other streptococci have no effect on red blood cells. Streptococcus
pyogenes, p. 487
MacConkey agar is differential as well as selective
(figure  4.11). In addition to its ingredients already men-
tioned, it contains lactose and a pH indicator.
Bacteria that ferment the sugar produce acid, which
turns the pH indicator pink. Colonies of lactose-
fermenting bacteria are pink on MacConkey agar,
whereas colonies of lactose-negative bacteria are
colorless. lactose, p. 31

Providing Appropriate
Atmospheric Conditions
To grow microorganisms on culture media in the laboratory,
appropriate atmospheric conditions must be provided.

Aerobic
When incubating most obligate aerobes and facultative anaerobes
on agar media, special atmospheric conditions are not required;
they can be incubated in air (approximately 20% O2). Broth cul-
tures of these organisms grow best when tubes or flasks containing FIGURE 4.12 Anaerobe Containers A disposable packet con-
the media are shaken, providing maximum aeration. tains chemicals that react with O2, thereby producing an anaerobic
Many medically important bacteria, including species of environment.
Neisseria and Haemophilus, grow best in aerobic atmospheres that ? Is the environment in a candle jar anaerobic?
 Part I Life and Death of Microorganisms 97

filled with an inert gas is used to add or remove items. Airtight

gloves allow researchers to handle items within the chamber.

Enrichment Cultures
An enrichment culture is used to isolate an organism present as
only a very small fraction of a mixed population. It does this by
providing conditions that preferentially enhance the growth of that
particular species in a broth (figure 4.14).
To enrich for a species, a sample is added to a broth medium
designed to favor the growth of the desired organism over others.
For example, if the target microbe can grow using atmospheric
nitrogen, then that element is left out of the medium. If it can use
an unusual carbon source such as phenol, then that compound is
added as the only carbon source. A selective agent such as bile
salts may also be added. The culture is then incubated in condi-
tions that preferentially promote the growth of the desired organ-
ism. As the microbes multiply, the relative concentration of the
FIGURE 4.13 Anaerobic Chamber The enclosed compartment target organism can increase dramatically. A pure culture can then
can be maintained as an anaerobic environment. A special port be obtained by streaking the enrichment onto an appropriate agar
(visible on the right side of this device) that can be filled with inert medium and selecting a single colony.
gas is used to add or remove items. The airtight gloves allow the
researcher to handle items within the chamber.
MicroAssessment 4.7
? Why would the chamber be preferable to an anaerobe container?
Culture media can be complex or chemically defined. Some media
contain additional ingredients that make them selective or differential.
Appropriate atmospheric conditions must be provided to grow
reducing agent such as sodium thioglycolate, which reduces O2 microbes in culture. An enrichment culture increases the relative
to water. In many cases, an O2-indicating dye is included as well. concentration of an organism growing in a broth.
The medium can be heated immediately before use to remove 18. Distinguish between complex and chemically defined media.
dissolved O2. reducing agent, p. 131 19. Describe two methods to create anaerobic conditions.
A more stringent method for working with anaerobes is to use 20. Would bacteria that cannot use lactose be able to grow on
an anaerobic chamber, an enclosed compartment maintained as MacConkey agar? +
an anaerobic environment (figure 4.13). A special port that can be

Inoculate and
Incubate incubate plate

Enriched sample is plated onto appropriate

Medium contains nutrients Sample that contains a variety Species of interest agar medium. A pure culture is obtained by
that few species other than of species, including the one of multiplies, whereas selecting a single colony of the species
the one of interest can use. interest, is added to the medium. others cannot. of interest.
FIGURE 4.14 Enrichment Culture Medium and incubation conditions favor the growth of the desired species over other microorganisms in
the same sample.
? How would you enrich for an organism that can use phenol as a carbon source?
98 Chapter 4 Dynamics of Prokaryotic Growth

Direct Microscopic Count

4.8 ■ Methods to Detect
One of the most rapid methods of determining the cell concen-
and Measure tration in a suspension is the direct microscopic count
Microbial Growth (figure 4.15). A liquid specimen is added to special glass slide
designed specifically for counting cells. The slide has a thin
Learning Outcome chamber that holds a known volume of liquid atop a micro-
15. Compare and contrast methods used for direct cell counts, scopic grid. The contents of the chamber can be viewed under
viable cell counts, measuring biomass, and detecting cell the light microscope, so the number of cells in a given volume
products. can be counted precisely. At least 10 million bacteria (107) per
milliliter are usually required for enough cells to be seen in the
A variety of techniques can be used to monitor microbial microscope field.
growth. Characteristics of common methods are summarized
in table 4.8.
Cell-Counting Instruments
A Coulter counter is an electronic instrument that counts cells in
Direct Cell Counts a suspension as they pass single file through a narrow channel
Direct cell counts are particularly useful for determining the total (figure 4.16). The suspending liquid must be an electrically con-
numbers of microbes, including those that cannot be grown in ducting fluid, because the machine counts the brief changes in
culture. Unfortunately, they generally do not distinguish between resistance that occur when non-conducting particles such as bac-
living and dead cells in the specimen. teria pass.

TABLE 4.8 Methods Used to Measure Microbial Growth

Method Characteristics and Limitations
Direct Cell Counts Used to determine total number of cells; counts include living and dead cells.

Direct microscopic count Rapid, but at least 107 cells/ml must be present to be effectively counted.

Cell-counting instruments Coulter counters and flow cytometers count total cells in dilute solutions. Flow cytometers can also be used to
count organisms to which fluorescent dyes or tags have been attached.

Viable Cell Counts Used to determine the number of viable microorganisms in a sample, but that number includes only those
that can grow in given conditions. Requires an incubation period of approximately 24 hours or longer.
Selective and differential media can be used to determine the number of specific microbial species.

Plate count Time-consuming but technically simple method that does not require sophisticated equipment. Generally used
only if the sample has at least 102 cells/ml.

Membrane filtration Concentrates microorganisms by filtration before they are plated; thus can be used to count cells in dilute
environments.

Most probable number Statistical estimation of likely cell number; it is not a precise measurement. Can be used to estimate numbers
of microorganisms in relatively dilute solutions.

Measuring Biomass Biomass can be correlated to cell number.

Turbidity Very rapid method; used routinely. A one-time correlation with plate counts is required in order to use
turbidity for determining cell number.

Total weight Tedious and time-consuming; however, it is one of the best methods for measuring the growth of filamentous
microorganisms.

Detecting Cell Products Used to detect growth, but not routinely used for quantitation.

Acid and Gas Production A pH indicator can be used to detect acid production. Gases can be trapped in an inverted Durham tube in a
tube of broth; a more sensitive method uses a fluorescent sensor that detects the slight decrease in pH that
accompanies CO2 production.
ATP Firefly luciferase catalyzes a light-emitting reaction when ATP is present.
 Part I Life and Death of Microorganisms 99

Cover glass resting on

supporting ridges
Counting grid
Counting chamber 00 0 0 25 89
Side view

Automatic counter
Sample spreads over counting
grid by capillary action.

Sample in
liquid

Bacterial cell
Electronic detector

Using a microscope, the cells in several large squares like the one shown
are counted and the results averaged. To determine the number of cells
per ml, that number must be multiplied by 1/volume (in ml) held in the
square. For example, if the square holds 1/1,250,000 ml, then the number FIGURE 4.16 A Coulter Counter This instrument counts cells as
of cells must be multiplied by 1.25 × 106 ml. they pass through a narrow channel.
FIGURE 4.15 Direct Microscopic Count The counting chamber ? Why do the microbial cells need to be suspended in an electrically
holds a known volume of liquid. The number of microbial cells in that conducting fluid for this method to work?
volume can be counted precisely.
? What is one disadvantage of doing a direct microscopic count to
determine the number of cells in a suspension?

A flow cytometer is similar in principle to a Coulter counter Plate Counts

except it measures light scattered by cells as they pass a laser. The Plate counts measure the number of viable cells in a sample by
instrument can be used to count either total cells or a specific taking advantage of the fact that an isolated microbial cell on a
subset that has been stained with a fluorescent dye or tag. fluo- nutrient agar plate will give rise to one colony. A simple count of
rescent dyes and tags, p. 49 the colonies determines how many cells were in the initial sample.
Plate counts are generally only done if a sample contains more
Viable Cell Counts than 100 organisms/ml. Otherwise, few if any cells will be trans-
Viable cell counts determine the number of cells capable of mul- ferred to the plates. In these situations, alternative methods give
tiplying. They are particularly valuable when working with sam- more reliable results.
ples such as food and water that contain too few microbes for a When counting colonies, the ideal number on a plate is
direct microscopic count. In addition, by using appropriate selec- between 30 and 300. Numbers outside of that range are more
tive and differential media, these methods can be used to count likely to be inaccurate. Samples usually contain many more cells
the cells of a particular microbial species. than that, so they generally must be diluted by a stepwise process
100 Chapter 4 Dynamics of Prokaryotic Growth

called serial dilution (figure  4.17). This is done using a sterile colony. When calculating CFUs, three things must be considered:
liquid called the diluent, often physiological saline (0.85% NaCl the number of colonies, the amount the sample was diluted before
in water). Dilutions are normally done in 10-fold increments, being plated, and the volume plated.
making the resulting math relatively simple.
Two techniques can be used to plate samples—spread-plate
and pour-plate (figure 4.18). In the spread-plate method, 0.1 to Membrane Filtration
0.2 ml of the diluted sample is transferred onto a plate of a solidi- Membrane filtration is used for liquid samples that contain rela-
fied agar medium. It is then spread over the surface of the agar tively few cells, as might occur in dilute environments such as
with a sterilized bent glass rod that resembles a miniature hockey natural waters. This method concentrates the microbes by filtra-
stick. In the pour-plate method, 0.1 to 1.0 ml of the diluted tion before they are plated. A known volume of liquid is passed
sample is transferred to a sterile Petri dish and then overlaid with through a sterile membrane filter that has a pore size small enough
a melted agar medium cooled to 50°C . At this temperature, agar to prevent microorganisms from passing through (figure  4.19).
is still liquid. The dish is then gently swirled to mix the microbial The filter is then placed on an appropriate agar medium and incu-
cells with the liquid agar. When the agar hardens, the individual bated. The number of colonies that form on the filter indicates the
cells become fixed in place; they form colonies when incubated. number of cells in the volume filtered.
Colonies that form on the surface will be larger than those embed-
ded in the medium.
In both methods, the plates are incubated and then the number Most Probable Number (MPN)
of colonies is counted. From that number, the concentration of The most probable number (MPN) is a method for estimating
colony-forming units (CFUs) in the sample can be determined. the concentration of cells in a specimen. The procedure uses a
This measure of viable cells accounts for the fact that microbial series of dilutions to determine the point at which subsequent dilu-
cells often attach to one another and then grow to form a single tions receive no cells.

Adding 1 ml of culture to 9 ml of diluent results in a 1:10 dilution.

Transfer 1 ml Transfer 1 ml Transfer 1 ml Transfer 1 ml

Original bacterial to 9 ml diluent to 9 ml diluent to 9 ml diluent to 9 ml diluent

culture 1:10 dilution 1:100 dilution 1:1,000 dilution 1:10,000 dilution
50,000 5,000 500 50 5
cells/ml cells/ml cells/ml cells/ml cells/ml

1 ml 1 ml 1 ml 1 ml 1 ml

Too many cells Too many cells Too many cells Between 30–300 Does not produce
produce too produce too produce too cells produces a enough colonies
many colonies many colonies many colonies countable plate. for a valid count.
to count. to count. to count.

FIGURE 4.17 Making Serial Dilutions To decrease the concentration of cells in a sample, 10-fold dilutions are often used.
? If the 1-ml sample had been added to 99 ml of diluent to make the first dilution (instead of 9 ml), how many cells would be in that particular
dilution?
 Spread-plate method

Solid agar

Incubate
Culture, diluted
as needed
Bacterial colonies
appear only on surface.
0.1–0.2 ml
Spread cells onto surface
of pre-poured solid agar.

Pour-plate method

0.1–1.0 ml

Melted cooled agar

Incubate

Some colonies appear on

Add melted cooled agar surface; many are below surface.
and swirl gently to mix.

FIGURE 4.18 Spread Plates and Pour Plates These techniques can be used for plate counts.
? Why are the results of plate counts expressed as the number of colony-forming units, rather than the number of cells?

FIGURE 4.19 Membrane Filtration This technique

concentrates microbial cells before they are plated.
? In what situation would membrane filtration be used
rather than a plate count?

Liquid
sample

Membrane
filter

A known volume of liquid is passed The membrane filter is placed on The number of colonies that grow on the
through a sterile membrane filter; the an appropriate agar medium and filter indicates the number of bacterial cells
filter retains bacterial cells. incubated. in the volume filtered.

101
102 Chapter 4 Dynamics of Prokaryotic Growth

Volume of Observation after incubation Number of positive Combination MPN Index/

inoculum (gas production noted) tubes in set of five of positives 100 ml

4-0-0 13

4-0-1 17
10 ml 4
4-1-0 17

4-1-1 21

4-1-2 26
+ – + + +
4-2-0 22

4-2-1 26

4-3-0 27
1 ml 3
4-3-1 33

4-4-0 34

– + + – + 5-0-0 23

5-0-1 30

5-0-2 40

0.1 ml 1 5-1-0 30

5-1-1 50

5-1-2 60
– – – + –

FIGURE 4.20 The Most Probable Number (MPN) Method In this example, three sets of five tubes containing the same growth medium
were prepared. Each set received the indicated amount of inoculum. After incubation, the presence or absence of gas in each tube was noted. The
results were then compared to an MPN table to get a statistical estimate of the concentration in the original sample of gas-producing bacteria
that could grow in the medium.
? If you were using a sample similar to the one illustrated, and all the tubes that received 10 ml had gas, but none that received 1.0 ml or 0.1 ml
did, what is the MPN index/100 ml?

To determine the MPN, three sets of three or five tubes containing and measures the percentage that reaches a light detector. That per-
a growth medium are prepared (figure 4.20). Each set receives a mea- centage is inversely proportional to the optical density. To use turbid-
sured amount of a sample such as water, soil, or food. The amount ity to estimate cell numbers, a one-time test must be done to determine
added is determined, in part, by the expected microbial concentration the correlation between optical density and cell concentration for the
in that sample. What is important is that the second set receives 10-fold specific organism and conditions under study. Once this correlation
less than the first, and the third set 100-fold less. In other words, each has been determined—generally using a direct microscopic count or
set is inoculated with an amount 10-fold less than the previous set. plate count to determine cell concentration—the turbidity measure-
After incubation, the presence or absence of turbidity or other indica- ment becomes a rapid and relatively accurate assay.
tion of growth is noted; the results are then compared against an MPN One limitation of using turbidity to measure biomass is that
table, which gives a statistical estimate of the cell concentration. the medium must contain a relatively high concentration of cells
to be cloudy. A solution containing 1 million bacteria (106) per ml
is still perfectly clear, and if it contains 10 times that amount, it is
Measuring Biomass barely turbid. It is important to remember that although a turbid
Instead of measuring the number of cells, the cell mass can be culture indicates that microbes are present, a clear solution does
determined. not guarantee their absence.

Turbidity Total Weight

The cloudiness or turbidity of a microbial suspension is proportional The total weight of a culture can be used to measure growth, but
to the concentration of cells, and is measured with a spectrophotom- the method is tedious and time-consuming. Because of this, total
eter (figure 4.21). This instrument shines light through a specimen weight is usually used to study only filamentous organisms that do
 Part I Life and Death of Microorganisms 103

Top scale = Percentage of

light that passes through

30 40 50 60 70
20 80
10 90
0 10
.4 .3 .2 0
8.7 .6 .5 .1 .0
1.9. 5
1.3 0

Bottom scale =
Optical density
(absorbance)

Light
detector

Light source
Dilute cell
(a) The cloudiness, or turbidity, of the liquid in the tube on the left suspension
is proportional to the concentration of cells.

30 40 50 60 70
20 80
10 90
0 10
.4 .3 .2 0
8.7 .6 .5 .1 .
1.9. 05
1.3 0

Concentrated cell
suspension
(c) The percentage of light that reaches the detector of the spectrophotometer
(b) A spectrophotometer is used to measure turbidity. is inversely proportional to the optical density.

FIGURE 4.21 Measuring Turbidity with a Spectrophotometer To use turbidity to estimate cell number, a one-time test must be done to
determine the correlation between optical density and cell concentration for the specific organism and conditions under study.
? Approximately how many bacterial cells must be in a suspension for it to be cloudy?

not readily separate into the individual cells necessary for a valid ATP
plate count. To measure the wet weight, cells in liquid culture are ATP can be detected by adding the firefly enzyme luciferase.
centrifuged and the liquid supernate removed. The weight of the The enzyme catalyzes a chemical reaction that uses ATP as an
resulting packed cell mass is proportional to the number of cells in energy source to produce light. This method is sometimes used
the culture. The dry weight can be determined by heating the cen- to assess the effectiveness of chemical treatments intended to
trifuged cells in an oven before weighing them. kill bacteria. Light is produced only if living organisms
remain.
Detecting Cell Products
Products of microbial growth can be used to detect the presence of MicroAssessment 4.8
microbes. Direct microscopic counts and cell-counting instruments generally
do not distinguish between living and dead cells. Plate counts
Acid and Gas Production determine the number of cells capable of multiplying; membrane
Microorganisms produce a variety of acids and, sometimes, gases filtration can be used to concentrate the sample. The most
as a result of their metabolism. Acids can be detected by includ- probable number estimates cell concentration. Turbidity of a
culture can be correlated to cell number. The total weight of
ing a pH indicator in the culture medium. Several pH indicators
a culture can be correlated to the number of cells present.
are available, and they differ in the pH value at which their color Microbial growth can be detected by the presence of cell products
changes. To detect gas production, inverted tubes (Durham tubes) such as acids, gases, and ATP.
can be used to trap gas bubbles in broth cultures. Clinical labs use 21. Why is an MPN an estimate rather than an accurate number?
more sensitive methods to detect the slight amounts of CO2 pro-
22. Water that accumulates in dirty bowls and dishes left in the
duced by bacteria growing in patient blood samples. One method sink often becomes cloudy over time. Besides suspended food
uses a fluorescent sensor to detect the slight decrease in pH that particles, what causes the turbidity? +
accompanies the production of CO2.
104 Chapter 4 Dynamics of Prokaryotic Growth

FUTURE CHALLENGES 4.1

Seeing How the Other 99% Lives
One of the biggest challenges for the future is cultures. Yet, over 99% of prokaryotes have with the DNA sequencing techniques dis-
to develop the methods to cultivate and study never been successfully grown in the labo- cussed in chapter 9, have made it easier to
a wider array of prokaryotes. Without these ratory. At the same time, when organisms study uncultivated microorganisms. This
microbes, humans and other animals would are removed from their natural habitat, and may well lead to a better understanding
not be able to exist. Yet, considering their especially when they are separated from of the diversity and the roles of microbes
importance, we still know very little about other organisms, their environment changes in our ecosystem. Scientists have learned
most species, including the relative contribu- drastically. Consequently, the study of pure a great deal since the days of Pasteur,
tions of each to such fundamental processes as cultures may not be the ideal for studying but most of the microbial world is still a
O2 generation and N2 and CO2 fixation. natural situations. mystery.
Much of our understanding of prokary- Technological advances such as flow
otic processes comes from work with pure cytometry and fluorescent labeling, along

Summary
4.1 ■ Principles of Prokaryotic Growth Continuous Culture
Most prokaryotes multiply by binary fission (figure  4.1). Microbial Microbes can be maintained in a state of continuous growth by using
growth is an increase in the number of cells in a population. a chemostat.
The time required for a population to double in number is the
generation time (table 4.1). 4.5 ■ Environmental Factors That Influence
Microbial Growth (table 4.2)
4.2 ■ Prokaryotic Growth in Nature
Temperature Requirements (figure 4.8)
Biofilms (figure 4.2)
Organisms can be grouped as psychrophiles, psychrotrophs,
Prokaryotes often live in a biofilm, a community encased in polysac- mesophiles, thermophiles, or hyperthermophiles based on their opti-
charides and other extracellular polymeric substances. mum growth temperatures.
Interactions of Mixed Microbial Communities Oxygen (O2) Requirements (table 4.3)
Prokaryotes often grow in close associations containing multiple differ- Organisms can be grouped as obligate aerobes, facultative anaerobes,
ent species. The metabolic activities of one organism often affects the obligate anaerobes, microaerophiles, or aerotolerant anaerobes based
growth of another. on their oxygen (O2) requirements.
4.3 ■ Obtaining a Pure Culture pH
Only an estimated 1% of prokaryotes have been grown in the laboratory. Organisms can be grouped as neutrophiles, acidophiles, or
alkaliphiles based on their optimum pH.
Growing Microorganisms on a Solid Medium
A single microbial cell deposited on a solid medium will multiply to Water Availability
form a visible colony (figure 4.4). Halophiles are adapted to live in high-salt environments.
The Streak-Plate Method (figure 4.5) 4.6 ■ Nutritional Factors That Influence Microbial Growth
The streak-plate method is used to isolate microorganisms in order to
obtain a pure culture. Required Elements (table 4.4)
The major elements make up cell constituents and include carbon,
Maintaining Stock Cultures nitrogen, sulfur, and phosphorus. Trace elements are required in very
Stock cultures can be stored on agar slants in the refrigerator, frozen, minute amounts.
or freeze-dried.
Growth Factors
4.4 ■ Prokaryotic Growth in Laboratory Conditions Microorganisms that cannot synthesize cell constituents such as amino
acids and vitamins require these as growth factors.
The Growth Curve (figure 4.6)
When grown in a closed system, a population of prokaryotic cells goes Energy Sources
through five phases: lag, exponential or log, stationary, death, and Organisms harvest energy either from sunlight or from chemical
prolonged decline. compounds.
Colony Growth Nutritional Diversity (table 4.5)
The position of a single cell within a colony markedly determines its Photoautotrophs use the energy of sunlight along with the carbon in
environment. the atmosphere to make organic compounds. Chemolithoautotrophs
 Part I Life and Death of Microorganisms 105

use inorganic compounds for energy and derive their carbon from 4.8 ■ Methods to Detect and
CO2. Photoheterotrophs use the energy of sunlight and obtain their Measure Microbial Growth (table 4.8)
carbon from organic compounds. Chemoorganoheterotrophs use
Direct Cell Counts
organic compounds for energy and as a carbon source.
Direct microscopic counts involve counting the number of cells
4.7 ■ Cultivating Prokaryotes in the Laboratory viewed through a microscope (figure 4.15). Both a Coulter counter and
a flow cytometer count cells as they pass through a minute aperture
General Categories of Culture Media (table 4.6) (figure 4.16).
A complex medium contains a variety of ingredients such as peptones
and extracts. A chemically defined medium is composed of precise Viable Cell Counts
mixtures of pure chemicals. Plate counts measure the number of viable cells by taking advan-
tage of the fact that an isolated cell will form a single colony
Special Types of Culture Media (figures 4.17, 4.18). Membrane filtration concentrates microbial
A selective medium inhibits organisms other than the one being cells by filtration; the filter is then incubated on an agar plate
sought. A differential medium contains a substance that certain micro- (figure 4.19). The most probable number (MPN) method is used a
organisms change in a recognizable way. series of inoculated tubes to statistical estimation of the cell con-
Providing Appropriate Atmospheric Conditions
centration (figure 4.20).
A candle jar provides increased CO2, which enhances the growth of Measuring Biomass
many medically important bacteria. Microaerophilic microbes are incu- Turbidity of a culture can be correlated with the number of cells; a
bated in a gastight container along with a packet that generates low O2 spectrophotometer is used to measure turbidity (figure 4.21). Wet weight
conditions. Anaerobes may be incubated in an anaerobe container or and dry weight are proportional to the number of cells in a culture.
an anaerobic chamber (figures 4.12, 4.13).
Detecting Cell Products
Enrichment Cultures (figure 4.14)
Products including acid, gas, and ATP can indicate growth.
An enrichment culture provides conditions in a broth that enhance the
growth of one particular organism in a mixed population.

Review Questions
Short Answer 2. Compared with their growth in the laboratory, bacteria in nature
1. Describe a detrimental and a beneficial effect of biofilms. generally grow
2. Define a pure culture. a) more slowly.
3. Explain what occurs during each of the five phases of growth. b) faster.
4. Explain how the environment of a colony differs from that of cells c) at the same rate.
growing in a liquid broth. 3. Cells are most sensitive to penicillin during which phase of the
5. List the five categories of optimum temperature, and describe a growth curve?
corresponding environment in which a representative might thrive. a) Lag b) Exponential c) Stationary
6. Why would botulism be a concern with canned foods? d) Death e) More than one of these.
7. Explain why O2-containing atmospheres kill some microbes. 4. Lactic acid is a primary metabolite. If a company wants to harvest
8. Explain why photoautotrophs are primary producers. this compound from a bacterial culture, the cells should be in
9. Distinguish between a selective medium and a differential which growth phase?
medium. a) Lag b) Exponential c) Stationary
10. If the number of microorganisms in lake water were determined d) Death e) More than one of these.
using both a direct microscopic count and a plate count, which 5. E. coli, a facultative anaerobe, is grown for 24 hours on the same
method would most likely give a higher number? Why? solid medium, but under two different conditions: one aerobic, the
other anaerobic. The size of the colonies would be
Multiple Choice a) the same under both conditions.
1. If there are 103 cells per ml at the middle of log phase, and the b) larger when grown under aerobic conditions.
generation time of the cells is 30 minutes, how many cells will c) larger when grown under anaerobic conditions.
there be 2 hours later?
a) 2 × 103 b) 4 × 103 c) 8 × 103
4 7
d) 1.6 × 10 e) 1 × 10
106 Chapter 4 Dynamics of Prokaryotic Growth

6. The generation time of a bacterium was measured at two different now must oversee its production. What are some factors you must
temperatures. Which results would be expected of a thermophile? consider if you need to grow 5,000-liter cultures of bacteria?
a) 20 minutes at 10°C; 220 minutes at 37°C 2. High-performance boat manufacturers know that microbes can
b) 220 minutes at 10°C; 20 minutes at 37°C collect on a boat, ruining its hydrodynamic properties. A boat-
c) no growth at 10°C; 20 minutes at 37°C manufacturing facility recently hired you to help with this problem
d) 20 minutes at 45°C; 220 minutes at 65°C because of your microbiology background. What strategies other
e) 220 minutes at 37°C; 20 minutes at 65°C than routine cleaning would you pursue to come up with a long-
term remedy for the problem?
7. Which of the following is false?
a) E. coli grows faster in nutrient broth than in glucose-salts Critical Thinking +
medium.
1. This figure shows a growth curve plotted on a non-logarithmic,
b) Organisms require nitrogen to make amino acids. or linear, scale. Compare this with figure 4.6. In both figures, the
c) Some eukaryotes can fix N2. number of cells increases dramatically during the log or exponen-
d) An organism that grows on ham is osmotolerant. tial phase. In this phase, the cell number increases more and more
e) Blood agar is used to detect hemolysis. rapidly (this effect is more apparent in the accompanying figure).
8. If the pH indicator were left out of MacConkey agar, the medium Why should the increase be speeding up?
would be
a) complex. b) differential. c) defined.
d) defined and differential. e) complex and differential.
9. A soil sample is placed in liquid and the number of bacteria in the
sample determined in two ways: (1) colony count and (2) direct
microscopic count. How would the results compare?
a) Methods 1 and 2 would give approximately the same results.
b) Many more bacteria would be estimated by method 1.

Number of cells

Log phase
c) Many more bacteria would be estimated by method 2.
d) Depending on the soil sample, sometimes method 1 would be
higher and sometimes method 2 would be higher.
10. The concentration of E. coli in a broth is between 104 and 106
cells per ml. To determine the precise number of living cells in the
sample, it would be best to
a) use a counting chamber.
b) plate out an appropriate dilution of the sample on nutrient agar.
c) determine cell number by using a spectrophotometer.
d) Any of these three methods would be satisfactory. Lag
phase
e) None of these three methods would be satisfactory.
Time
Applications
1. You are a microbiologist working for a pharmaceutical company 2. In question 1, how would the curve appear if the availability of
and discover a new metabolite that can serve as a medication. You nutrients were increased?
5 Control of Microbial Growth
KEY TERMS
Antiseptic A disinfectant non-
toxic enough to be used on skin.
Aseptic Technique Procedures
Pasteurization Brief heat
treatment that reduces the number
of spoilage organisms and destroys
disease-causing microbes.
that minimize the chance of
unwanted microbes being Preservation Process of inhibiting
accidentally introduced. microbial growth to delay spoilage.
Bactericidal Kills bacteria. Sterilant A chemical that destroys
all microbes.
Bacteriostatic Prevents the growth
of, but does not kill, bacteria. Sterile Completely free of all viable
microbes; an absolute term.
Disinfectant A chemical that
destroys many microbes. Sterilization The destruction or
removal of all microbes through
Germicide Kills microbes.
physical or chemical means.

U
p until the late nineteenth century, patients undergoing
even minor surgeries were at great risk of developing fatal
infections due to unsanitary medical practices and hospital
conditions. Physicians did not know that their hands could pass
diseases from one patient to the next. Nor did they understand
Medical settings warrant a high level of microbial control. that airborne microscopic organisms could infect open wounds.
Fortunately, today’s modern hospitals use strict procedures to
avoid microbial contamination, allowing most surgeries to be
A Glimpse of History performed with relative safety.
The British Medical Journal stated that the British physician Joseph Microbial growth affects more than our health. Manufacturers
Lister (1827–1912) “saved more lives by the introduction of his system of a wide variety of goods recognize that microorganisms can
than all the wars of the 19th century together had sacrificed.” Lister reduce product quality. Their growth can lead to undesirable
revolutionized surgery by introducing methods that prevent wounds from changes in the safety, appearance, taste, odor, or function of prod-
becoming infected. ucts ranging from food to lumber.
Impressed with Pasteur’s work on fermentation (said to be caused This chapter covers methods to destroy, remove, and inhibit
by “minute organisms suspended in the air”), Lister wondered if “minute microbial growth on inanimate objects and some body surfaces.
organisms” might be responsible also for the pus that formed in wounds. Most of these approaches can damage all forms of life. In contrast,
He then experimented by applying carbolic acid, a toxic compound, antibiotics and other antimicrobial medications are specifically
directly onto damaged tissues and found it prevented infections. Carbolic
toxic to microbes, which is why they are so valuable in treating
acid wound dressings then became standard in his practice, and he took
infectious diseases; they will be discussed in chapter 20.
pride in the fact that his patients no longer developed gangrene. His work
also provided impressive evidence for the germ theory of disease, even
though microorganisms specific for various diseases were not identified 5.1 ■ Approaches to Control
for another decade.
Later, Lister improved his methods even further by using surgi- Learning Outcomes
cal procedures that excluded bacteria from wounds. These procedures 1. Using the appropriate terminology, describe the principles
included sterilizing instruments before use and maintaining a clean envi- of sterilization, disinfection, pasteurization, decontamination,
ronment in the operating room. Doing this was preferable to killing the sanitization, and preservation.
bacteria after they entered wounds because they avoided the toxic effects 2. Compare and contrast the methods used to control microbial growth in
of the disinfectant on the wound. daily life, healthcare settings, microbiology laboratories, food and food
Lister was knighted in 1883 and subsequently became a baron and a production facilities, water treatment facilities, and other industries.
member of the House of Lords.

107
108 Chapter 5 Control of Microbial Growth

The processes used to control microorganisms are either physical laboratories, foods and food production facilities, water treatment
or chemical, though a combination of both can be used. Physical facilities, and other industries.
methods include heat treatment, irradiation, filtration, and mechan-
ical removal (washing). Chemical methods use any of a variety of Daily Life
antimicrobial chemicals. The method chosen depends on the cir- Washing and scrubbing with soaps and detergents is a sufficient
cumstances and level of control required. level of microbial control in routine situations. Soap itself gener-
ally does not destroy many organisms; it simply aids in the
mechanical removal of microbes, including most pathogens, as
Principles of Control well as dirt, organic material, and some skin cells. Regular hand-
Sterilization is the removal or destruction of all microorganisms washing and bathing does not adversely affect the beneficial nor-
and viruses on or in a product. Microbes can be removed by filtra- mal skin microbiota, which reside more deeply on underlying
tion, or destroyed using heat, certain chemicals, or irradiation. layers of skin cells and in hair follicles. Other methods used to
Destruction of microorganisms means they cannot be “revived” to control microorganisms in daily life include cooking foods, clean-
multiply even when transferred from the sterilized product to an ing surfaces, and refrigeration.
ideal growth medium. A sterile item is one that is free of microbes,
including endospores and viruses. It is important to note, however, MicroByte
Thorough handwashing with soap and water is the most important
that the term sterile does not consider prions. These infectious
step in stopping the spread of many infectious diseases.
protein particles are not destroyed by standard sterilization proce-
dures. endospores, p. 67 prions, p. 328
Disinfection is the elimination of most or all pathogens on or Hospitals and other Healthcare Facilities
in a material. In practice, the term generally implies the use of Minimizing the numbers of microorganisms in healthcare settings
antimicrobial chemicals. Unlike sterilization, disinfection suggests is particularly important because of the danger of healthcare-
that some living microbes may remain. Disinfectants are antimi- associated infections. Patients in healthcare facilities, particularly
crobial chemicals used for disinfecting inanimate objects. They are hospitals, are often more susceptible to infectious agents because
toxic to many forms of life, and therefore biocides (bio means of their weakened condition. In addition, patients may undergo
“life,” and cida means “to kill”). They are typically used in a man- invasive procedures such as surgery, which cuts the intact skin
ner that targets microorganisms and viruses, however, so they are that would otherwise help prevent infection. Finally, pathogens
often called germicides. They are also described as bactericidal, are more likely to be found in healthcare settings because of the
meaning they kill bacteria. Antiseptics are antimicrobial chemicals high concentration of patients with infectious disease. These
non-toxic enough to be used on skin or other body tissue. These are patients can shed pathogens in their feces, urine, respiratory drop-
routinely used to decrease bacterial numbers on skin before inva- lets, or other body secretions. Because of this, healthcare facilities
sive procedures such as surgery. pathogen, p. 6 must be extremely careful to control microorganisms. Nowhere is
Pasteurization is a brief heat treatment that reduces the num- this more important than in the operating rooms, where instru-
ber of spoilage organisms and destroys pathogens. Foods and ments used in invasive procedures must be sterile to avoid intro-
inanimate objects can be pasteurized. ducing even normally harmless microbes into deep body tissue
Decontamination is a process used to reduce the number of where they could easily cause infection.  healthcare-associated infec-
pathogens to a level considered safe to handle. The treatment can tions, p. 449
be as simple as thorough washing, or it may involve the use of heat Prions are a relatively new concern for healthcare facilities.
or disinfectants. Fortunately, disease caused by these agents is thought to be
Sanitization generally implies a process that substantially exceedingly rare, less than 1 case per 1 million persons per year.
reduces the microbial population to meets accepted health stan- Healthcare facilities, however, must take special precautions when
dards. Most people also expect a sanitized object to look clean. handling tissue that may be contaminated with prions, because
Note that this term does not indicate any specific level of control. these infectious particles are very difficult to destroy.
Preservation is the process of delaying spoilage of foods or
other perishable products. One way to do this is to adjust storage Microbiology Laboratories
conditions to slow microbial growth. Alternatively, chemical pre- Microbiology laboratories routinely work with microbial cultures
servatives can be added. These are bacteriostatic, meaning they and consequently must use rigorous methods to control microor-
inhibit the growth of bacteria but do not kill them. ganisms. To work with pure cultures, all media and instruments
that contact the culture must first be sterilized to avoid contami-
nating the culture with environmental microbes. All materials
Situational Considerations used to grow microorganisms must again be treated before dis-
Methods used to control microbial growth vary greatly depending posal to avoid contamination of workers and the environment. The
on the situation and level of control required (figure 5.1). Control use of specific methods to prevent microorganisms from contami-
measures adequate for routine circumstances of daily life might nating an environment is called aseptic technique. Although all
not be sufficient for situations such as hospitals, microbiology microbiology laboratory personnel must use these measures, those
 Part I Life and Death of Microorganisms 109

(a)

(b) (c)

(d) (e)
FIGURE 5.1 Situations That Warrant Different Levels of Microbial Control (a) Daily home life; (b) foods and food production facilities;
(c) water treatment facilities; (d) hospitals; (e) other industries.
? Why would a hospital require a more stringent level of microbial control than daily home life?

who work with known pathogens must be even more careful. microorganisms. These are known as biosafety levels (BSLs) and
aseptic technique, p. 85 range from BSL-1 (for work with microbes not known to cause
The Centers for Disease Control and Prevention (CDC) has disease) to BSL-4 (for work with deadly pathogens for which no
established precaution guidelines for laboratories working with vaccine or specific treatment exists).
110 Chapter 5 Control of Microbial Growth

Foods and Food Production Facilities Selection of an effective antimicrobial procedure is complicated
Foods and other perishable products retain their quality longer by the fact that every procedure has disadvantages that limit its
when contaminating microbes are destroyed, removed, or inhib- use. An ideal, multipurpose, non-toxic method simply does not
ited. Heat treatment is the most common and reliable method used exist. The ultimate choice depends on many factors including the
to kill microbes, but it can alter the flavor and appearance of the type and number of microbes, environmental conditions, risk for
products. Irradiation can be used to destroy microbes, and chemi- infection, and the composition of the item.
cal additives can be used to prevent their growth, but the risk of
toxicity must always be a concern. Because of this, the Food and
Drug Administration (FDA) regulates these options. Type of Microbes
Food-processing facilities need to keep surfaces relatively One of the most critical considerations in selecting an antimicro-
free of microorganisms to avoid contamination. If machinery used bial procedure is the microbial population. Products contaminated
to grind meat is not cleaned properly, for example, it can create an with microbes highly resistant to killing require a more rigorous
environment in which bacteria multiply, eventually contaminating treatment. Highly resistant microbes include:
large quantities of product.
■ Bacterial endospores. The endospores of Bacillus, Clostridium,
and related genera are the most resistant form of life typically
Water Treatment Facilities encountered. Only extreme heat or chemical treatment ensures
Water treatment facilities need to ensure that drinking water is free their complete destruction. endospores, p. 67
of pathogenic microbes. Chlorine has traditionally been used to ■ Protozoan cysts and oocysts. Cysts and oocysts are stages
disinfect water, saving hundreds of thousands of lives by prevent- in the life cycle of certain intestinal protozoan pathogens such
ing the spread of waterborne illnesses such as cholera. Chlorine as Giardia lamblia and Cryptosporidium parvum. These
and other disinfectants, however, can react with naturally occur- disinfectant-resistant forms are excreted in the feces of infected
ring chemicals in the water to form disinfection by-products animals, including humans, and can cause diarrheal disease if
(DBPs). Some of these have been linked to long-term health risks. ingested. Unlike endospores, they are easily destroyed by boiling.
In addition, certain pathogens, particularly Cryptosporidium Cryptosporidium parvum, p. 604 Giardia lamblia, p. 602
parvum, a cause of diarrhea, can survive traditional disinfection ■ Mycobacterium species. The waxy cell walls of mycobacteria
procedures. To address these problems, water treatment regula- make them resistant to many chemical treatments. Because of
tions now require facilities to minimize the level of both DBPs and this, stronger, more toxic chemicals must be used to disinfect
C. parvum in treated water. Cryptosporidium parvum, p. 604 environments that may contain Mycobacterium tuberculosis,
the cause of tuberculosis. tuberculosis, p. 502
Other Industries ■ Pseudomonas species. These common environmental organ-
Many diverse industries have specialized concerns regarding isms are not only resistant to some disinfectants, but in some
microbial growth. Manufacturers of pharmaceuticals, cosmetics, cases actually grow in them. Pseudomonas species can cause
deodorants, or any other product that will be ingested, injected, or serious healthcare-associated infections. Pseudomonas infec-
tions, p. 554
applied to the skin must avoid microbial contamination that could
affect the product’s quality or safety. ■ Naked viruses. Viruses such as poliovirus that lack a lipid
envelope are more resistant to disinfectants. Conversely,
MicroAssessment 5.1 enveloped viruses, such as HIV, tend to be very sensitive to
these chemicals. naked viruses, p. 306 enveloped viruses, p. 306
The methods used to control microbial growth depend on the situation
and the level of control required.
1. How is sterilization different from disinfection?
2. Why would water treatment facilities be concerned about
Number of Microorganisms
disinfection by-products? The time it takes for heat or chemicals to kill a microbial popula-
3. Why would the term sterilization not encompass prions? + tion is dictated in part by the number of cells present. It takes more
time to kill a large population than it does to kill a small popula-
tion, because only a fraction of organisms die during a given time
5.2 ■ Selection of an interval. For example, if 90% of a bacterial population is killed
during the first 3 minutes, then approximately 90% of those
Antimicrobial Procedure remaining will be killed during the next 3 minutes, and so on.
Removing organisms by washing or scrubbing can minimize the
Learning Outcome
time necessary to sterilize or disinfect a product.
3. Explain why the type and number of microbes, environmental In the commercial canning industry, the decimal reduction
conditions, risk for infection, and composition of the item influence
time, or D value, is the time required for killing 90% of a bacterial
the selection of an antimicrobial procedure.
population under specific conditions (figure 5.2). The temperature
 Part I Life and Death of Microorganisms 111

■ Semicritical instruments. These come into contact with

108
Log decrease of 1 mucous membranes, but do not penetrate body tissue; they
107 include gastrointestinal endoscopes and endotracheal tubes.
Number of surviving cells (logarithmic scale)
Semicritical instruments must be free of all viruses and veg-
106 etative bacteria. The few endospores that may remain pose
little risk for infection because mucous membranes are effec-
Logarithmic
105 killing tive barriers against their entry into deeper tissue.
■ Non-critical instruments and surfaces. These come into
104 contact only with unbroken skin so they pose little risk for
infection. Countertops, stethoscopes, and blood pressure cuffs
103 are examples of non-critical items.
Log
decrease
102 of 1
Composition of the Item
101
Some sterilization and disinfection procedures are inappropriate
“D” “D”
for certain types of material. For example, heat treatment can dam-
1 age many types of plastics and other materials. Irradiation pro-
0 30 60 90 120 150 vides an alternative to heat, but the process damages some types
Time (min) of plastics. Moist heat (such as boiling water) and liquid chemical
disinfectants cannot be used to treat moisture-sensitive material.
FIGURE 5.2 D Value The D value is the time it takes to reduce the
population by 90%.
? Based on this graph, what is the approximate D value for the
organism?
MicroAssessment 5.2
The types and numbers of microorganisms initially present,
environmental conditions, the potential risks associated with use
of the process is indicated by a subscript, for example, D121. A one of the item, and the composition of the item must all be considered
D process reduces the number of cells by one exponent. Thus, if the when determining which sterilization or disinfection procedure to
D value for an organism is 2 minutes, then it would take 4 minutes employ.
(2 D values) to reduce a population of 100 (102) cells to only one 4. Describe three groups of microorganisms that are resistant to
(100) survivor. It would take 20 minutes (10 D values) to reduce a certain chemical treatments.
population of 1010 cells to only one survivor. 5. Why do critical instruments need to be sterile, whereas
semicritical instruments need only be free of viruses and
vegetative bacteria?
6. Would it be safe to say that if all bacterial endospores had been
Environmental Conditions killed, then all other medically important microorganisms had
Dirt, grease, and body fluids such as blood can interfere with heat also been killed? +
penetration and the action of chemical disinfectants. This is
another reason why it is important to thoroughly clean items
before disinfection or sterilization.
Temperature and pH influence microbial death rates. A solu-
tion of sodium hypochlorite (household bleach), for example, can 5.3 ■ Using Heat to Destroy
kill a suspension of M. tuberculosis at a temperature of 55°C in
half the time it would take if the suspension were held at 50°C. Microorganisms
The hypochlorite solution is even more effective at a low pH. and Viruses
Learning Outcomes
Risk for Infection 4. Compare and contrast pasteurization, sterilization using
pressurized steam, and the commercial canning process.
To guide the selection of germicidal procedures, medical instru-
5. Explain the drawbacks and benefits of using dry heat rather than
ments are categorized according to their risk for transmitting
moist heat to kill microorganisms.
infectious agents. Those that pose greater threats require more
rigorous germicidal procedures. The categories are:
Heat treatment is one of the most useful methods of microbial
■ Critical instruments. These come into direct contact with control because it is reliable, safe, relatively fast and inexpensive,
body tissues; they include needles and scalpels. Critical and does not introduce potentially toxic substances into materials.
instruments must be sterile. Some heat-based methods sterilize the product, whereas others
112 Chapter 5 Control of Microbial Growth

decrease the number of microbes. Table 5.1 summarizes the char- Pasteurization

acteristics of heat treatment and other physical methods of control. Louis Pasteur developed the brief heat treatment we now call pas-
teurization as a way to prevent spoilage of wine. Today, pasteuri-
zation is still used to destroy heat-sensitive spoilage organisms in
Moist Heat foods and beverages, increasing the product’s shelf life without
Moist heat destroys microbes by irreversibly denaturing their pro- significantly altering its quality. More importantly, pasteurization
teins. Examples of moist heat treatment include boiling, pasteuri- is widely used to destroy pathogens in milk and juices, protecting
zation, and pressurized steam. consumers from diseases such as tuberculosis, brucellosis, salmo-
nellosis, and typhoid fever. food spoilage, p. 756
Today, most pasteurization is high-temperature–short-time
Boiling (HTST). With this treatment, milk is heated to 72°C and held for
Boiling (100°C at sea level) easily destroys most microorganisms 15 seconds, but the parameters must be adjusted to the product.
and viruses. Because of this, drinking water that might be con- For example, ice cream is rich in fats, so it is pasteurized at 82°C
taminated during floods or other emergency situations should be for about 20 seconds.
boiled for at least 5 minutes. Boiling is not a method of steriliza- Shelf-stable boxed juices and milk, as well as the single-
tion, however, because endospores can survive the process. serving containers of half-cream served in restaurants, are treated

TABLE 5.1 Physical Methods Used to Destroy Microorganisms and Viruses

Method Characteristic Use

Moist Heat Denatures proteins. Relatively fast, reliable, safe, Widely used.
and inexpensive.
Boiling Boiling for 5 minutes destroys most microorganisms Boiling for at least 5 minutes can be used to treat
and viruses; a notable exception is endospores. drinking water.
Pasteurization Significantly decreases the numbers of heat-sensitive Milk is pasteurized by heating it to 72°C for 15 seconds.
microorganisms, including spoilage microbes and Juices are also routinely pasteurized.
pathogens (except sporeformers).
Pressurized Typical treatment is 121°C/15 psi for 15 minutes or longer, Widely used to sterilize microbiological media,
steam a process that destroys endospores. laboratory glassware, surgical instruments, and other
(autoclaving) items that steam can penetrate. The canning process
renders foods commercially sterile.
Dry Heat
Incineration Burns cell components to ashes. Flaming of wire inoculating loops. Also used to destroy
medical wastes and contaminated animal carcasses.
Dry heat ovens Destroys cell components and denatures proteins. Less Laboratory glassware is sterilized by heating at 160°C
efficient than moist heat, requiring longer times and to 170°C for 2 to 3 hours. Powders, oils, and other dry
higher temperatures. materials are also sterilized in ovens.
Filtration Filter retains microbes while letting the suspending fluid
or air pass through small holes.
Filtration of Various pore sizes are available; 0.2 μm is commonly used Used for beer and wine, and to sterilize some heat-
fluids to remove bacteria. sensitive medications.
Filtration of air HEPA filters are used to remove microbes that have Used in biological safety cabinets, specialized hospital
a diameter of 0.3 μm or greater. rooms, and airplanes. Also used in some vacuum cleaners
and home air purification units.
Radiation Type of cell damage depends on the wavelength of the
radiation.
Ionizing Destroys DNA and possibly damages cytoplasmic Used to sterilize heat-sensitive materials including
radiation membranes. Produces reactive molecules that damage medical equipment, disposable surgical supplies, and
other cell components. Items can be sterilized even drugs such as penicillin. Also used to destroy microbes in
after packaging. spices, herbs, and approved types of produce and meats.
Ultraviolet Damages DNA. Penetrates poorly. Used to destroy microbes in the air and drinking water,
radiation and to disinfect surfaces.
High Pressure Treatments of 130,000 psi are thought to denature proteins Used to extend the shelf life of certain commercial food
and alter the permeability of the cell. Products retain color products such as guacamole.
and flavor.
 Part I Life and Death of Microorganisms 113

Exhaust valve to Valve to sterilization with a higher temperature for a shorter time can be
remove steam control steam used. To destroy prions, autoclaving at a temperature of 132°C for
after sterilization to chamber Pressure gauge
Safety
1 hour is thought to be effective.
valve Autoclaving is consistently effective in sterilizing most
Door objects, if done correctly. The temperature and pressure gauges of
the autoclave should both be monitored to ensure proper operating
Steam conditions. It is also critical that steam enter items to displace air.
Because of this, long, thin containers should be placed on their
sides, and containers should never be closed tightly.
Tape that contains a heat-sensitive indicator, which turns
Air
black at a high temperature, should be attached to items before
Jacket autoclaving. This provides a visual signal that the items have been
processed (figure 5.4a). A changed indicator, however, does not
mean that the object is sterile, only that it has been heated.
Biological indicators are used to ensure that an autoclave is work-
Thermometer ing properly (figure 5.4b). A tube containing the heat-resistant endo-
Trap spores of Geobacillus stearothermophilus can be placed near the
Pressure center of an item. After that item is autoclaved, the endospores are
regulator
mixed with a growth medium by manually crushing a container within
Steam supply the tube. If the medium changes color during incubation, microbial
growth occurred, indicating an unsuccessful process.
FIGURE 5.3 Autoclave Steam first travels in an enclosed layer,
or jacket, surrounding the chamber. It then enters the autoclave,
displacing the air downward and out through a port in the bottom The Commercial Canning Process
of the chamber. Commercial canning uses an industrial-sized autoclave (called a
? Why is autoclaving more effective than boiling? retort). The canning process is designed to ensure that endospores
of Clostridium botulinum are destroyed. This is critical because
surviving spores can germinate in canned foods such as vegetables
using ultra-high-temperature (UHT) processing. This destroys and meats. The resulting vegetative cells can grow in low-acid
all microorganisms that can grow under normal storage condi- anaerobic conditions and produce botulinum toxin, one of the
tions, so it is referred to as “ultra-pasteurization.” The UHT pro- most potent toxins known. botulism, p. 652

cess for milk requires rapidly heating the milk to 140°C, holding In destroying endospores of C. botulinum, the canning pro-
it at that temperature for a few seconds, and then quickly cooling cess also kills all other organisms that grow under normal storage
it. The product is then aseptically packaged in sterile containers. conditions. Canned foods are commercially sterile, meaning that
the endospores of some thermophiles may survive. These are usu-
MicroByte ally not a concern, however, because they grow only at tempera-
Clothes can be pasteurized by regulating the temperature in a tures well above those of normal storage.
washing machine.

Sterilization Using Pressurized Steam

Heat- and moisture-tolerant items such as surgical instruments,
most microbiological media, and reusable glassware are sterilized
using autoclaves. Water in a chamber in the autoclave is heated to
form steam, causing the pressure in the chamber to increase
(figure 5.3). The higher pressure, in turn, increases the tempera-
ture at which steam forms. Steam at atmospheric pressure never
exceeds 100°C, but steam at an additional 15 psi (pounds per
square inch) is 121°C, a temperature that kills even endospores.
The pressure itself plays no direct role in the killing.
Typical conditions for sterilization are 15 psi and 121°C for (a) (b)
15 minutes. Longer treatment is necessary for large volumes
FIGURE 5.4 Indicator Used in Autoclaving (a) Chemical
because it takes more time for heat to completely penetrate the indicators. The pack on the left has been autoclaved. Diagonal marks
substance. For example, 4 liters of nutrient broth in a flask takes on the tape have turned black, indicating that the object was exposed
longer to sterilize than the same volume distributed into tubes. to heat. (b) Biological indicators. Following incubation, a change of
Other autoclave conditions can be used for specific purposes. color to yellow indicates growth of the endospore-forming organism.
When rapid processing is important, such as when sterile instru- ? Why would a biological indicator be better than other indicators
ments must always be available in an operating room, flash to determine if the sterilization procedure was effective?
114 Chapter 5 Control of Microbial Growth

Several factors dictate the time and temperature of the can- Some materials cannot withstand heat treatment. For these items,
ning process. First, as discussed earlier, the higher the tempera- other physical methods including filtration, irradiation, and high-
ture, the shorter the time needed to kill all organisms. Second, the pressure treatment can be used to destroy or remove microbes.
higher the concentration of organisms, the longer the heat treat-
ment required to kill them all. To provide a wide safety margin,
the commercial canning process is designed to reduce a popula- Filtration
tion of 1012 C. botulinum endospores to only one spore. In other Recall that membrane filtration, used to determine the number of
words, it is a 12 D process. It is virtually impossible for a food to bacteria in a liquid medium, retains bacteria while allowing the
have this many endospores. fluid to pass through. That same principle can be used to physi-
cally remove microbes from liquids or air. membrane filtration,
p. 100
Dry Heat
Dry heat is not as efficient as moist heat in killing microbes, and Filtration of Fluids
therefore requires longer times and higher temperatures. For Filtration is used extensively to remove organisms from heat-
example, 200°C for 90 minutes of dry heat has the killing equiva- sensitive fluids such as sugar solutions, beer, and wine. Specially
lent of 121°C for 15 minutes of moist heat. designed filtration units are also used by backpackers and campers
Incineration burns the cell components to ashes. In microbiol- to remove Giardia cysts and bacteria from water.
ogy laboratories, the wire loops continually reused to transfer Paper-thin membrane filters or microfilters have micro-
bacterial cultures are sterilized by flaming—heating them in a scopic pores that allow liquid to flow through while trapping
flame until they are red hot. Alternatively, they can be heated to particles too large to pass (figure  5.5). A vacuum is commonly
the same point in a benchtop incinerator designed for this purpose. used to help pull the liquid through the filter; alternatively, pres-
Incineration is also used to destroy medical wastes and contami- sure may be applied to push the liquid. The filters are made of
nated animal carcasses. nitrocellulose or other polymers and come in a variety of different
Temperatures achieved in hot air ovens destroy cell compo- pore sizes, extending below the dimensions of the smallest known
nents and irreversibly denature proteins. Glass Petri dishes and viruses. Pore sizes smaller than necessary should be avoided,
glass pipettes are sterilized in ovens with non-circulating air at however, because they slow the flow. Filters with a pore size of
temperatures of 160°C to 170°C for 2 to 3 hours. Ovens with a fan 0.2 micrometers (μm) are commonly used to remove bacteria.
that circulates the hot air can sterilize in a shorter time because Depth filters trap material within thick porous filtration
they transfer heat more efficiently. Powders, oils, and other dry material such as cellulose fibers. They have complex passages that
material are also sterilized in hot ovens. retain microorganisms while letting the suspending fluid pass
through the small holes. The diameter of the passages is often
MicroAssessment 5.3 much larger than that of microbes, but the electrical charges on the
Moist heat such as boiling water destroys most microbes. walls help hold the microbial cells. cellulose, p. 32
Pasteurization significantly reduces the numbers of heat-sensitive
organisms. Autoclaves use pressurized steam to achieve high
temperatures that kill microbes, including endospores. The
commercial canning process is designed to destroy the endospores of
Clostridium botulinum. Dry heat takes longer than moist heat to kill
microbes.
7. Why is it important that the commercial canning process
destroys the endospores of Clostridium botulinum?
Filter
8. How does incineration destroy microbes?
9. Would pasteurization destroy endospores? +

Flask
5.4 ■ Using Other Physical
Methods to Remove Vacuum
pump
or Destroy Microbes Sterilized
fluid
Learning Outcomes FIGURE 5.5 Sterilization of Fluids Using a Membrane Filter
6. Describe how depth filters, membrane filters, and HEPA filters are The liquid to be sterilized flows through the filter on top of the flask
used to remove microorganisms. in response to a vacuum produced in the flask by means of a pump.
Scanning electron micrograph (5,000×) shows a membrane filter
7. Compare and contrast the use of gamma radiation, ultraviolet retaining cells of Pseudomonas.
radiation, and microwaves for destroying microorganisms.
? Why would a pore size smaller than necessary be a poor choice?
 Part I Life and Death of Microorganisms 115

Filtration of Air forms, whereas Gram-negative bacteria such as Salmonella and

High-efficiency particulate air (HEPA) filters remove nearly all Pseudomonas species are among the most susceptible. Two
airborne particles 0.3 μm or larger. These filters are used for keep- important forms of ionizing radiation are gamma rays (emitted
ing microorganisms out of specialized hospital rooms designed for from decaying radioisotopes such as cobalt-60) and X rays.
patients extremely susceptible to infection. The filters are also reactive oxygen species, p. 90 radioisotope, p. 19

used in biological safety cabinets in which laboratory personnel Ionizing radiation is used extensively to sterilize heat-
work with dangerous airborne pathogens such as Mycobacterium sensitive materials including medical equipment, disposable sur-
tuberculosis. A continuous stream of incoming and outgoing air gical supplies, and drugs such as penicillin. Irradiation can
flows through HEPA filters to hold microbes within the cabinet. generally be carried out after packaging.
The cabinets are also used to protect samples from contamination. The number of microbes destroyed by irradiating a food prod-
uct depends on the dose applied. Treatments designed to sterilize
MicroByte food can cause unwanted flavor changes, however, which limits
HEPA filters are used in passenger aircraft (to remove microbes their usefulness. More commonly, food is irradiated as a method
from the recirculated cabin air) and in vacuum cleaners. of eliminating pathogens and decreasing the numbers of spoilage
organisms. For example, it can be used to kill pathogens such as
Salmonella species in poultry with little or no change in taste of
Radiation the product.
Radio waves, microwaves, visible and ultraviolet (UV) light rays, In the United States, irradiation has been used for many years
X rays, and gamma rays are all examples of electromagnetic to control microorganisms on spices and herbs. The FDA has also
radiation. This is a form of energy that travels in waves and has approved irradiation of fruits, vegetables, and grains (to control
no mass; the amount of energy is related to the wavelength, which insects), pork (to control parasites), and poultry, beef, lamb, and
is the distance from crest to crest (or trough to trough) of a wave. pork (to control bacterial pathogens). Many consumers refuse to
The wavelength is inversely proportional to the frequency, the accept irradiated products, even though the FDA and officials of
number of waves per second. Radiation that has short waves, and the World Health and the United Nations Food and Agriculture
therefore high frequency, has more energy than that which has Organizations have endorsed the technique. Some people mistak-
long waves and low frequency. The full range of wavelengths is enly believe that irradiated products are radioactive. Others think
called the electromagnetic spectrum (figure 5.6). that irradiation-induced toxins or carcinogens are present in food,
even though available scientific evidence indicates that consump-
Ionizing Radiation tion of irradiated food is safe. Another argument raised against
irradiation is that it will allow people to ignore other important
Ionizing radiation has enough energy to remove electrons from
food-safety practices. Irradiation, however, is intended to comple-
atoms. This harms cells directly by destroying DNA and damag-
ment, not replace, proper food-handling procedures by producers,
ing cytoplasmic membranes. It also causes indirect damage, react-
processors, and consumers.
ing with O2 to produce reactive oxygen species (ROS). Bacterial
endospores are among the most radiation-resistant microbial
Ultraviolet Radiation
Ultraviolet light in wavelengths of approximately 220 to 300 nm
Wavelength (nm) destroys microbes by damaging their DNA. Actively multiplying
200 300 400 500 600 700
organisms are the most easily killed, whereas bacterial endospores
Bactericidal are the most UV-resistant.
Ultraviolet light is used extensively to destroy microbes in the
Ultraviolet (UV) light Visible light
air and drinking water and to disinfect surfaces. It penetrates
poorly, however, which limits its use. Even a thin film of grease
Ionizing radiation on the UV bulb or material covering microbial cells can make it
Gamma less effective. Likewise, it cannot be used to destroy microbes in
X rays UV Infrared Microwaves Radio waves
rays solid substances or turbid liquids. Because most types of glass and
10 –5 10 –3 1 10 3 10 6 10 9 10 12
plastic screen out ultraviolet radiation, UV light is most effective
Wavelength (nm) when used at close range against exposed microorganisms. It must
be used carefully because UV rays can also damage the skin and
Increasing energy
One wavelength eyes and promote the development of skin cancers.
Crest

Trough Microwaves
Increasing wavelength
Microwaves do not affect microorganisms directly, but the heat
FIGURE 5.6 The Electromagnetic Spectrum Visible wavelengths they generate can be lethal. It is important to remember, however,
include the colors of the rainbow. that microwave ovens often heat food unevenly, so even heat-
? Which types of radiation are ionizing? sensitive cells can sometimes survive the process.
116 Chapter 5 Control of Microbial Growth

High Pressure procedures originally defined by the EPA. Germicides are grouped
according to their potency:
High-pressure processing is used to decrease the number of
microbes in commercial food products, such as guacamole, with- ■ Sterilants. These can destroy all microorganisms, includ-
out using high temperatures. The process, which uses pressures of ing endospores and viruses. They are also called sporocides.
up to 130,000 psi, is thought to destroy microbes by denaturing Destruction of endospores usually requires a 6- to 10-hour
proteins and altering cell permeability. High-pressure treated treatment. Sterilants are used to treat heat-sensitive critical
products keep the color and flavor associated with fresh foods. instruments such as scalpels.
■ High-level disinfectants. These destroy all viruses and veg-
MicroAssessment 5.4
etative microorganisms, but they do not reliably kill endospores.
Filters can be used to remove microorganisms and viruses from Most are simply sterilants used for short time periods, not long
liquids and air. Gamma radiation can be used to sterilize products enough to ensure endospore destruction. They are used to treat
and to decrease the number of microorganisms in foods. Ultraviolet
semicritical instruments such as gastrointestinal endoscopes.
light can be used to disinfect surfaces and air. Microwaves do not kill
microbes directly but destroy them by the heat they generate. Extreme ■ Intermediate-level disinfectants. These destroy all vegeta-
pressure can kill microorganisms. tive bacteria including mycobacteria, fungi, and most, but not
10. What is the difference between the mechanism of a depth filter all, viruses. They do not kill endospores even with prolonged
and that of a membrane filter? exposure. This group of germicides is used to disinfect non-
11. How does ultraviolet light kill microorganisms? critical instruments such as stethoscopes.
12. Why could sterilization by gamma irradiation be carried out ■ Low-level disinfectants. These destroy fungi, vegetative
even after packaging? + bacteria except mycobacteria, and enveloped viruses. They do
not kill endospores, nor do they always destroy naked viruses.
Intermediate-level and low-level disinfectants are also called
5.5 ■ Using Chemicals to Destroy general-purpose disinfectants. In hospitals, they are used for
disinfecting furniture, floors, and walls.
Microorganisms and Viruses
To perform properly, germicides must be used strictly according
Learning Outcomes to the manufacturer’s instructions, especially those for dilution,
8. Describe the difference between sterilants, high-level temperature, and the amount of time they must be in contact with
disinfectants, intermediate-level disinfectants, and low-level the object being treated. It is extremely important to thoroughly
disinfectants. clean objects before they are treated to both decrease the number
9. Describe five important factors to consider when selecting an of microbes present and remove organic material that might inter-
appropriate germicidal chemical. fere with the activity of the chemical.
10. Compare and contrast the characteristics and use of alcohols,
aldehydes, biguanides, ethylene oxide gas, halogens, metals,
ozone, peroxygens, phenolic compounds, and quaternary
Selecting the Appropriate
ammonium compounds as germicidal chemicals. Germicidal Chemical
Selecting the appropriate germicide is a complex decision. Some
Germicidal chemicals can be used to disinfect and, in some cases, points to consider include:
sterilize. Most react irreversibly with proteins, DNA, cytoplasmic
■ Toxicity. Germicides are at least somewhat toxic to humans
membranes or viral envelopes. Their mechanisms of action, how-
ever, are often poorly understood. They are generally less reliable and the environment. Therefore, the benefit of disinfecting or
than heat but useful for treating large surfaces and heat-sensitive sterilizing an item or surface must be weighed against the risks
items. Some are sufficiently non-toxic to be used as antiseptics. associated with using the chemical. For example, the risk of being
exposed to pathogens in a hospital warrants using the most effec-
tive germicides, even considering the potential risks of their use.
Potency of Germicidal The microbiological risks associated with typical household and
Chemical Formulations office situations, however, may not justify using many of those
same chemicals. To encourage the use of less toxic options, the
Numerous different germicidal chemicals are marketed for medi-
EPA developed a “Design for the Environment” pilot program
cal and industrial use under a variety of trade names. Frequently,
that allows manufacturers to place a specially designed logo on
they contain more than one antimicrobial chemical as well as other
products considered the least hazardous.
chemicals, such as buffers that can affect their antimicrobial activ-
ity. In the United States, the FDA is responsible for regulating ■ Activity in the presence of organic matter. Hypochorite
chemicals that can be used to process medical devices in order to (bleach) and many other germicides react with organic matter,
ensure they work as claimed. Most other chemical disinfectants losing their effectiveness in doing so. Chemicals such as phe-
are considered pesticides and, as such, are regulated by the nolics, however, tolerate the presence of some organic matter.
Environmental Protection Agency (EPA). To be registered with ■ Compatibility with the material being treated. Items
either the FDA or EPA, manufacturers of germicidal chemicals such as electrical equipment often cannot withstand liquid
must document the potency of their products using testing chemical germicides, and so gaseous alternatives must be
 Part I Life and Death of Microorganisms 117

employed. Likewise, corrosive germicides such as hypochlo- The stocks are simply diluted according to the manufacturer’s
rite often damage some metals and rubber. instructions before use. Some have a limited shelf life once
■ Residue. Many chemical germicides leave a toxic or corro- prepared.
sive residue. If such a germicide is used to treat an item, the ■ Environmental risk. Germicides that retain their antimicro-
residue must be removed by washing the item. bial activity after use can interfere with wastewater treatment
■ Cost and availability. Some germicides are less expensive systems. The activity of those germicides must be neutralized
and more readily available than others. Hypochlorite can eas- before disposal. wastewater treatment, p. 736
ily be purchased in the form of household bleach. In contrast,
ethylene oxide gas is not only expensive, but requires a special Classes of Germicidal Chemicals
chamber, which affects the cost and practicality. Germicides are represented in a number of chemical families.
■ Storage and stability. Some germicides are sold as concen- Each type has characteristics that make it more or less appropriate
trated stock solutions, decreasing the required storage space. for specific uses (table 5.2).

TABLE 5.2 Chemicals Used in Sterilization, Disinfection, and Preservation of Non-Food Substances
Chemical (examples) Characteristics Uses

Alcohols (ethanol and isopropanol) Easy to obtain and inexpensive. Rapid Aqueous solutions of alcohol are used as
evaporation limits their contact time. antiseptics to clean skin in preparation
for procedures that break intact skin, and
as disinfectants for treating instruments.
Aldehydes (glutaraldehyde, Capable of destroying all microbes. Irritating to Glutaraldehyde and orthophthalaldehyde are
orthophthalaldehyde, and the respiratory tract, skin, and eyes. used to sterilize medical instruments. Formalin
formaldehyde) is used in vaccine production and to preserve
biological specimens.
Biguanides (chlorhexidine) Relatively low toxicity, destroys a wide range of Chlorhexidine is widely used as an antiseptic
microbes, adheres to and persists on skin and in soaps and lotions, and is often impregnated
mucous membranes. into catheters and surgical mesh.
Ethylene Oxide Gas Easily penetrates hard-to-reach places and Commonly used to sterilize medical devices.
fabrics and does not damage moisture-sensitive
material. It is toxic, explosive, and may be
carcinogenic.
Halogens (chlorine and iodine) Chlorine solutions are inexpensive and readily Solutions of chlorine are widely used to
available; however, organic compounds and disinfect inanimate objects, surfaces, drinking
other impurities neutralize the activity. Some water, and wastewater. Tincture of iodine and
forms of chlorine may react with organic iodophores can be used as disinfectants or
compounds to form toxic chlorinated products. antiseptics.
Iodine is more expensive than chlorine and
does not reliably kill endospores.
Metals (silver) Most metal compounds are too toxic to be Silver sulfadiazine is used in topical dressings to
used medically. prevent infection of burns. Silver nitrate drops
can be used to prevent eye infections caused
by Neisseria gonorrhoeae in newborns. Some
metal compounds are used to prevent microbial
growth in industrial processes.
Ozone This unstable form of oxygen readily breaks Used to disinfect drinking water and
down to an ineffective form. wastewater.
Peroxygens (hydrogen peroxide Readily biodegradable and less toxic than Hydrogen peroxide is used to sterilize
and peracetic acid) traditional alternatives. The effectiveness of containers for aseptically packaged juices and
hydrogen peroxide as an antiseptic is limited milk. Peracetic acid is widely used to disinfect
because the enzyme catalase breaks it down. and sterilize medical devices.
Peracetic acid is a more potent germicide than
hydrogen peroxide.
Phenolic Compounds (triclosan and Wide range of activity, reasonable cost, Triclosan is used in a variety of personal care
hexachlorophene) remains effective in the presence of detergents products, including toothpastes, lotions, and
and organic contaminants, leaves an active deodorant soaps. Hexachlorophene is highly
antimicrobial residue. effective against Staphylococcus aureus, but its use
is limited because it can cause neurological damage.
Quaternary Ammonium Compounds Non-toxic enough to be used on food Widely used to disinfect inanimate objects and
(benzalkonium chloride and preparation surfaces. Inactivated by anionic to preserve non-food substances.
cetylpyridinium chloride) soaps and detergents.
118 Chapter 5 Control of Microbial Growth

Alcohols disease. Adverse side effects of chlorhexidine are rare, but severe
Aqueous solutions of 60% to 80% ethyl or isopropyl alcohol allergic reactions have been reported.
quickly kill vegetative bacteria and fungi. They do not, however,
reliably destroy bacterial endospores and some naked viruses. Ethylene Oxide
Alcohol denatures essential proteins such as enzymes, and dam- Ethylene oxide is an extremely useful gaseous sterilizing agent
ages lipid membranes. Proteins are more soluble and denature that destroys all microbes, including endospores and viruses, by
more easily in alcohol mixed with water, which is why the aque- reacting with proteins. As a gas, it penetrates well into fabrics,
ous solutions are more effective than pure alcohol. equipment, and implantable devices such as pacemakers and arti-
Alcohol solutions are commonly used as antiseptics to clean ficial hips. It is particularly useful for sterilizing heat- or moisture-
skin before procedures such as injections that break intact skin. In sensitive items such as electrical equipment, pillows, and
addition, the Centers for Disease Control and Prevention recom- mattresses. Many disposable laboratory items, including plastic
mends that alcohol-based hand sanitizers be used routinely by Petri dishes and pipettes, are also sterilized with ethylene oxide.
healthcare personnel as a means to protect patients. A special chamber that resembles an autoclave is used to
Alcohol solutions are also used as disinfectants for treating sterilize items with ethylene oxide. This allows careful control of
instruments and surfaces. They are relatively non-toxic and inex- factors such as temperature, relative humidity, and ethylene oxide
pensive and do not leave a residue. Unfortunately, they evaporate concentration, all of which influence the effectiveness of the gas.
quickly, which limits their contact time and, consequently, their Because ethylene oxide is explosive, it is generally mixed with a
germicidal effectiveness. In addition, they can damage rubber, non-flammable gas such as carbon dioxide. Under these carefully
some plastics, and other material. controlled conditions, objects can be sterilized in 3 to 12 hours.
Antimicrobial chemicals such as iodine are sometimes dis- The toxic ethylene oxide must then be eliminated from the treated
solved in alcohol. These alcohol-based solutions, called tinctures, material using heated forced air for 8 to 12 hours. This is impor-
can be more effective than the corresponding aqueous solutions. tant because the chemical irritates tissues and has a persistent
antimicrobial effect, which, in the case of Petri dishes and other
Aldehydes items used for growing bacteria, is unacceptable.
The aldehydes glutaraldehyde, orthophthalaldehyde (OPA), and Ethylene oxide is mutagenic and may be carcinogenic.
formaldehyde destroy microorganisms and viruses by inactivating Studies have shown a slightly increased risk of malignancies in
proteins and nucleic acids. A 2% solution of alkaline glutaralde- long-term users of the gas.
hyde is one of the most widely used liquid chemical sterilants for
treating heat-sensitive medical items. Immersion in this solution Halogens
for 10 to 12 hours destroys all microbes, including endospores and Chlorine and iodine are common disinfectants that are thought to
viruses. Vegetative cells can be killed with soaking times as short act by reacting with proteins and other essential cell components.
as 10 minutes. Glutaraldehyde is toxic, however, so treated items
must be thoroughly rinsed with sterile water before use. Chlorine Chlorine destroys all types of microorganisms and vi-
Orthophthalaldehyde is a relatively new type of disinfectant ruses but is too irritating to skin and mucous membranes to be used
that provides an alternative to glutaraldehyde. It requires shorter as an antiseptic. Chlorine-releasing compounds such as sodium
processing times and is less irritating to eyes and nasal passages, hypochlorite can be used to disinfect waste liquids, swimming pool
but it stains proteins gray, including those of the skin. water, instruments, and surfaces, and at much lower
Formaldehyde is used as a gas or as formalin, an aqueous concentrations, to disinfect drinking water.
37% solution. It is an extremely effective germicide that kills most Chlorine solutions are inexpensive, readily
microbes quickly. Formalin is used to kill bacteria and inactivate available disinfectants (figure  5.7). An effec-
viruses for use as vaccines. It has also been used to preserve bio- tive solution can easily be made by diluting
logical specimens. Formaldehyde releases irritating vapors and is liquid household bleach (5.25% sodium
a suspected carcinogen, limiting its use. hypochlorite) 1:100 in water, resulting in
a solution of 500  ppm (parts per mil-
Biguanides lion) chlorine. This concentration is
Chlorhexidine, the most effective of a group of chemicals called several hundred times the amount
biguanides, is extensively used in antiseptic products. It stays on
skin and mucous membranes, is of relatively low toxicity, and
destroys a wide range of microbes, including vegetative bacteria,
fungi, and some enveloped viruses. Chlorhexidine is an ingredient
in many products including antiseptic skin creams, disinfectants,
and mouthwashes. Chlorhexidine-impregnated catheters and FIGURE 5.7 Chlorine in the Form of
implanted surgical mesh are used in medical procedures. Even Household Bleach
tiny chips that slowly release chlorohexidine have been devel- ? Why should bleach concentrations
oped; these are inserted into periodontal pockets to treat gum higher than necessary be avoided?
 Part I Life and Death of Microorganisms 119

PERSPECTIVE 5.1
Contamination of an Operating Room by a Bacterial Pathogen
A patient with burns infected with Pseudomonas
aeruginosa was taken to the operating room
to have the wounds cleaned and dead tissue
removed. After the procedure was completed,
samples of various surfaces in the room were
cultured to determine the extent of contamina-
tion. P.  aeruginosa was recovered from all
parts of the room. Figure 1 shows how readily
and extensively an operating room can become
contaminated by an infected patient. Operating
rooms and other patient care rooms must be
thoroughly cleaned after use, in a process
known as terminal cleaning.

FIGURE 1 Contaminated Operating Room An operating

room in which tissue infected with Pseudomonas aeruginosa
was removed from a burn patient. Reddish areas indicate places
where P. aeruginosa was recovered following the surgical
procedures.

required to kill most pathogens, but usually necessary for fast, Iodine Iodine, unlike chlorine, does not reliably kill endospores,
reliable effects. In situations when excessive organic material is but it can be used as a disinfectant. It is used as a tincture, or more
present, a 1:10 dilution of bleach may be required. This is because commonly as an iodophore, in which the iodine is linked to a car-
chlorine readily reacts with organic compounds and other impuri- rier molecule that releases free (unbound) iodine slowly. Iodo-
ties in water, disrupting its germicidal activity. The use of high phores are not as irritating to the skin as tincture of iodine nor are
concentrations, however, should be avoided when possible, they as likely to stain. Iodophores used as disinfectants contain
because chlorine is both corrosive and toxic. Diluted bleach dete- more free iodine (30 to 50 ppm) than do those used as antiseptics
riorates over time, so fresh solutions should be prepared regularly. (1 to 2 ppm). Stock solutions must be strictly diluted according to
More stable forms of chlorine, including sodium dichloroisocyan- the manufacturer’s instructions because dilution affects the
urate and chloramines, are often used in hospitals. amount of free iodine available.
Chlorine is used at very low levels to disinfect drinking water; Surprisingly, some Pseudomonas species survive in the con-
the amount needed depends on the amount of organic material in centrated stock solutions of iodophores. The reasons are unclear,
the water. The presence of organic compounds is also a problem but it could be due to inadequate levels of free iodine in concen-
because chlorine can react with some organic compounds to form trated solutions, because iodine may be released from the carrier
trihalomethanes—disinfection by-products that are potential car- only with dilution. Pseudomonas species also can form biofilms,
cinogens. Although chlorination of drinking water kills many which are less permeable to chemicals. Healthcare-associated
pathogens, it is important to remember that the levels used are not infections can result if a Pseudomonas-contaminated iodophore is
effective against Cryptosporidium parvum oocysts and Giardia unknowingly used to disinfect instruments. biofilm, p. 84
lamblia cysts. Cryptosporidium parvum, p. 604 Giardia lamblia, p. 602
Chlorine dioxide (ClO2) is a strong oxidizing agent increas-
ingly being used as a disinfectant and sterilant. It has an advantage Metal Compounds
over chlorine-releasing compounds in that it does not react with Metal compounds kill microorganisms by combining with sulfhy-
organic compounds to form trihalomethanes or other toxic chlori- dryl groups (—SH) of enzymes and other proteins, thereby inter-
nated products. Compressed chlorine dioxide gas, however, is fering with their function. High concentrations of most metals are
explosive and liquid solutions decompose readily, so it must be too toxic to human tissue to be used medically.
generated on-site. It is used to treat drinking water, wastewater, Silver is one of the few metals still used as a disinfectant.
and swimming pools. oxidizing agent, p. 131 Creams containing silver sulfadiazine, a combination of silver and
120 Chapter 5 Control of Microbial Growth

a sulfa drug, are applied topically to prevent infection of second- be used to sterilize items in less than 1 hour. It is effective in
and third-degree burns. Commercially available bandages with the presence of organic compounds, leaves no residue, and can
silver-containing pads can be used on minor scalds, cuts, and be used on a wide range of materials. It has a sharp, strong
scrapes. For many years, doctors were required by law to add odor, however, and like other oxidizing agents, it irritates the
drops of another silver compound, 1% silver nitrate, into the eyes skin and eyes.
of newborns. This was to prevent ophthalmia neonatorum, an eye
infection caused by Neisseria gonorrhoeae, acquired from infected Phenolic Compounds (Phenolics)
mothers during the birth process. Drops of antibiotics have now Phenol (carbolic acid) is important historically because it was one
largely replaced use of silver nitrate because they are less irritating of the earliest disinfectants (see A Glimpse of History), but it has
to the eye and more effective against another genitally acquired an unpleasant odor and irritates the skin. Phenolics are derivatives
pathogen, Chlamydia trachomatis. Neisseria gonorrhoeae, p.  622
of phenol that have greater germicidal activity. Because they are
Chlamydia trachomatis, p. 625
so effective, more dilute and therefore less irritating solutions can
Compounds of mercury, tin, arsenic, copper, and other metals be used. Phenolics are the active ingredients in Lysol.
were once widely used as preservatives in industrial products and Phenolics destroy cytoplasmic membranes of microorgan-
to prevent microbial growth in recirculating cooling water. Their isms and denature proteins. They kill most vegetative bacteria
extensive use resulted in serious pollution of natural waters, which and, in high concentrations (from 5% to 19%), many can kill
has prompted strict controls. Mycobacterium species. They do not, however, reliably inactivate
all groups of viruses. The major advantages of phenolic com-
MicroByte
In emergencies, drinking water can be disinfected by adding two pounds include their wide range of activity, reasonable cost, and
drops of plain bleach to a liter of water; let sit for 30 minutes before ability to remain effective in the presence of detergents and
ingesting. organic contaminants. They also leave an active antimicrobial
residue, which in some cases is desirable.
Some phenolics, such as triclosan and hexachlorophene, are
Ozone sufficiently non-toxic to be used in soaps and lotions. Triclosan is
Ozone (O3), an unstable form of oxygen, is a powerful oxidizing widely used as an ingredient in a variety of personal care products
agent. It decomposes quickly, however, so it must be generated such as deodorant soaps, lotions, and toothpaste. Hexachlorophene
on-site, usually by passing air or O2 between two electrodes. has substantial activity against Staphylococcus aureus, the leading
Ozone is used as an alternative to chlorine for disinfecting drink- cause of wound infections, but high levels have been associated
ing water and wastewater. with symptoms of neurotoxicity. Although once widely used in
over-the-counter products, antiseptic skin cleansers containing
Peroxygens hexachlorophene are now available only with a prescription.
Hydrogen peroxide and peracetic acid are powerful oxidizing Staphylococcus aureus, p. 524

agents that can be used as sterilants under controlled conditions.

They are readily biodegradable and, in normal concentrations of Quaternary Ammonium Compounds (Quats)
use, appear to be less toxic than the traditional alternatives (ethyl- Quaternary ammonium compounds (quats) are cationic (positively
ene oxide and glutaraldehyde). charged) detergents non-toxic enough to be used to disinfect food
preparation surfaces. Like all detergents, quats have both a
Hydrogen Peroxide The effectiveness of hydrogen peroxide charged hydrophilic region and an uncharged hydrophobic region.
(H2O2) as a germicide depends in part on whether it is used on Because of this, they reduce the surface tension of liquids and help
living tissue or an inanimate object. This is because all cells that wash away dirt and organic material, facilitating the mechanical
use aerobic metabolism, including the body’s tissue cells, produce removal of microbes from surfaces. Unlike most common house-
catalase, the enzyme that inactivates hydrogen peroxide by break- hold soaps and detergents, however, which are anionic (negatively
ing it down to water and O2. Thus, when a solution of 3% hydro- charged) and repelled by the negatively charged microbial cell
gen peroxide is applied to a wound, our cellular enzymes quickly surface, quats are attracted to the cell surface. They react with
break it down. When the same solution is used on an inanimate membranes, destroying many vegetative bacteria and enveloped
surface, however, it overwhelms the relatively low concentration viruses. They are not effective, however, against endospores,
of catalase produced by microbial cells. catalase, p. 90 mycobacteria, or naked viruses.
Hydrogen peroxide is particularly useful as a disinfectant Quats are economical, effective, and widely used to disin-
because it leaves no residue and does not damage stainless steel, fect clean inanimate objects and preserve non-food substances.
rubber, plastic, or glass. Hot solutions are commonly used in the The ingredients of many personal care products include quats
food industry to produce commercially sterile containers for asepti- such as benzalkonium chloride or cetylpyridinium chloride.
cally packaged juices and milk. Vapor-phase hydrogen peroxide is They also enhance the effectiveness of some other disinfec-
more effective than liquid solutions and can be used as a sterilant. tants. Cationic soaps and organic material such as gauze, how-
ever, can neutralize their activity. In addition, Pseudomonas, a
Peracetic Acid Peracetic acid is an even more potent germi- troublesome cause of healthcare-associated infections, resists
cide than hydrogen peroxide. A 0.2% solution of peracetic acid, the effects of quats and can even grow in solutions preserved
or a combination of peracetic acid and hydrogen peroxide, can with them.
 Part I Life and Death of Microorganisms 121

MicroAssessment 5.5 which are carcinogens—by the metabolic activities of intestinal

bacteria or during cooking.
Germicidal chemicals can be used to disinfect and, in some cases,
sterilize, but they are less reliable than heat. They are especially MicroByte
useful for destroying microorganisms and viruses on heat-sensitive Swiss cheese naturally contains propionic acid, and cranberries have
items and large surfaces. benzoic acid.
13. Describe four factors to consider when selecting a germicide.
14. Explain why it is essential to dilute iodophores properly.
15. Why would a heavy metal be a more serious pollutant than most Low-Temperature Storage
organic compounds? +
Refrigeration inhibits the growth of many pathogens and spoilage
microorganisms by slowing or stopping critical enzyme reactions.
Psychrotrophic and some psychrophilic organisms, however, can
5.6 ■ Preservation of Perishable grow at refrigeration temperatures. psychrophiles, p. 89
Freezing preserves foods and other products by stopping
Products microbial growth. The ice crystals that form kill some of the
microbial cells, but those that survive can grow and spoil foods
Learning Outcome once thawed.
11. Compare and contrast chemical preservatives, low-temperature
storage, and reducing the available water as methods to preserve
perishable products. Reducing the Available Water
Salting and drying decrease the availability of water in food below
Preventing or slowing the growth of microorganisms extends the
the limits required for growth of most microbes. The high-solute
shelf life of products such as food, medicines, deodorants, cosmet-
environment also causes plasmolysis, which damages microbial cells
ics, and contact lens solutions. Preservatives are often added to
(see figure 4.9). water availability, pp. 91, 749 plasmolysis, p. 91
these products to prevent or slow the growth of microbes inevita-
bly introduced from the environment. Other common methods of
slowing microbial growth include low-temperature storage such
Adding Sugar or Salt
as refrigeration or freezing, and reducing available water. These
methods are particularly important in preserving foods. food
Sugar and salt draw water out of cells, dehydrating them. High
spoilage, p. 756 food preservation, p. 759
concentrations of these solutes are added to many foods as preser-
vatives (figure 5.8). For example, fruit is made into jams and

Chemical Preservatives
Some of the germicidal chemicals described in the previous sec-
tion can be used to preserve non-food items. For example, mouth-
washes often contain a quaternary ammonium compound,
cosmetics may contain thimerosal, and leather belts may be
treated with one or more phenol derivatives. Food preservatives,
however, must be non-toxic for safe ingestion.
Benzoic, sorbic, and propionic acids are weak organic acids
sometimes added to foods such as bread, cheese, and juice to
prevent microbial growth. In acidic conditions, these chemicals
affect cell membrane functions. Although this action inhibits the
growth of many microbes, the low pH itself prevents the growth
of most bacteria. Therefore, the primary benefit of adding the
preservatives is that they inhibit fungi, which otherwise grow at
acidic pH.
Nitrate and its reduced form, nitrite, serve a dual purpose in
processed meats. From a microbiological viewpoint, their most
important function is to inhibit the germination of endospores and
subsequent growth of Clostridium botulinum. If low levels of
nitrate or nitrite are not added to cured meats such as bologna, ham,
bacon, and smoked fish, C. botulinum may grow and produce
deadly botulinum toxin. At higher concentrations than required for
preservation, nitrate and nitrite react with myoglobin in the meat to
form a stable pigment that gives a desirable pink color associated
with fresh meat. The preservatives pose a potential hazard, how- FIGURE 5.8 Preserved Jams and Jellies
ever, because they can be converted to nitrosamines—some of ? How does a high concentration of sugar function as a preservative?
122 Chapter 5 Control of Microbial Growth

jellies by adding sugar, and fish and meats are cured by soaking Although drying stops microbial growth, it does not reliably
them in salty water, or brine. It is important to note, however, that kill bacteria and fungi in or on foods. For example, numerous
the food-poisoning bacterium Staphylococcus aureus can grow in cases of salmonellosis have been traced to dried eggs. Eggshells
relatively high-salt conditions. Staphylococcus aureus, p. 524 and even egg yolks may be heavily contaminated with Salmonella
species from the gastrointestinal tract of the hen. To prevent the
Drying Food transmission of such pathogens, some states have laws requiring
Foods that are dried often have added salt, sugar, or chemical dried eggs to be pasteurized before they are sold.
preservatives. For example, meat jerkies usually have added salt
and sometimes sugar.
Lyophilization (freeze-drying) is widely used for preserving MicroAssessment 5.6
foods such as coffee, milk, meats, and vegetables. In the process Preservation techniques slow or halt the growth of microorganisms to
of freeze-drying, the food is first frozen and then dried in a vac- delay spoilage.
uum. When water is added to the lyophilized material, it reconsti- 16. What is the most important function of nitrate in cured meat?
tutes. The quality of the reconstituted product is often much better 17. How do high concentrations of sugar and salt preserve foods?
than that of products dried using ordinary methods. The light
18. Preservation by freezing is sometimes compared to drying.
weight and stability without refrigeration of freeze-dried foods Why would this be so? +
make them popular with hikers.

FUTURE CHALLENGES 5.1

Too Much of a Good Thing?
In our complex world, the solution to one chal- The issues surrounding the excessive use antibiotic resistance. By using antimicrobial
lenge may inadvertently lead to the creation of antimicrobial chemicals are complicated. chemicals indiscriminately, we may eventually
of another. Scientists have long been pursu- On the one hand, there is no question that make these useful tools obsolete. Excessive use
ing less toxic alternatives to many traditional some microorganisms cause disease. Even of disinfectants may even be contributing to the
biocidal chemicals. For example, glutaralde- those that are not harmful to human health problems of antibiotic resistance. Disinfectant-
hyde has now largely replaced the more toxic can be troublesome because their metabolic resistant bacteria sometimes overproduce
formaldehyde, chlorhexidine is generally used end products can ruin the quality of perish- efflux pumps that expel otherwise damaging
in place of hexachlorophene, and gaseous able products. Based on that information, it chemicals, including antibiotics, from the cell.
alternatives to ethylene oxide are now being seems wise to destroy or inhibit the growth Thus, by overusing disinfectants, we may be
sought. Meanwhile, ozone and hydrogen per- of microorganisms whenever possible. The inadvertently increasing antibiotic resistance.
oxide, which are both readily biodegradable, role of microorganisms in our life, however, Another concern is over the misguided
may eventually replace glutaraldehyde. While is not that simple. Our bodies actually har- belief that “non-toxic” or “biodegradable”
these less toxic alternatives are better for bor a greater number of microbial cells than chemicals cause no harm, and the common
human health and the environment, their wide- human cells, and this normal microbiota plays notion that “if a little is good, more is even
spread acceptance and use may be unwittingly an important role in maintaining our health. better.” For example, concentrated solutions
contributing to an additional problem—the Excessive use of antiseptics or other antimi- of hydrogen peroxide, though biodegradable,
overuse and misuse of germicidal chemicals. crobials may actually predispose a person to can cause serious damage, even death, when
Many products, including soaps, toothbrushes, infection by damaging the normal microbiota. used improperly. Other chemicals, such as
and even clothing and toys are marketed with An even more worrisome concern is that chlorhexidine, can elicit severe allergic reac-
the claim of containing antimicrobial ingredi- overuse of disinfectants and other germicidal tions in some people.
ents. Already bacterial resistance to some of chemicals will select for microorganisms that As less toxic germicidal chemicals are
the chemicals included in these products has are more resistant to those chemicals, a situ- developed, people must be educated on the
been reported. ation similar to our current problems with appropriate use of these alternatives.

Summary
5.1 ■ Approaches to Control
Situational Considerations (figure 5.1)
The methods used to destroy or remove microbes and viruses can be
physical, such as heat treatment, irradiation, and filtration, or chemical. Situations encountered in daily life, hospitals, microbiology labora-
tories, food production facilities, water treatment facilities, and other
Principles of Control industries warrant different degrees of microbial control.
A variety of terms are used to describe antimicrobial agents and processes.
 Part I Life and Death of Microorganisms 123

5.2 ■ Selection of an Antimicrobial Procedure High Pressure

Type of Microbe
High pressure is thought to destroy microorganisms by denaturing pro-
teins and altering the permeability of the cell.
One of the most critical considerations in selecting a method of destroy-
ing microorganisms and viruses is the type of microbial population
5.5 ■ Using Chemicals to Destroy
thought to be present on or in the product.
Microorganisms and Viruses
Number of Microorganisms
Potency of Germicidal Chemical Formulations
The amount of time it takes for heat or chemicals to kill a popula-
Germicides are grouped according to their potency as sterilants, high-
tion of microorganisms is dictated in part by the number of cells
level disinfectants, intermediate-level disinfectants, or low-level
initially present. Microbial death generally occurs at a constant
disinfectants.
rate. The D value, or decimal reduction time, is the time it takes
to kill 90% of a population of bacteria under specific conditions Selecting the Appropriate Germicidal Chemical
(figure 5.2). Factors that must be included in the selection of an appropriate ger-
Environmental Conditions micidal chemical include toxicity, residue, activity in the presence of
organic matter, compatibility with the material being treated, cost and
Factors such as pH and presence of organic materials influence micro-
availability, storage and stability, and ease of disposal.
bial death rates.
Classes of Germicidal Chemicals (table 5.2)
Risk for Infection
Solutions of 60% to 80% ethyl or isopropyl alcohol in water rap-
Medical instruments are categorized as critical, semicritical, and
idly kill vegetative bacteria and fungi by coagulating enzymes
non-critical according to their risk of transmitting infectious agents.
and other essential proteins, and by damaging lipid membranes.
Composition of the Item Glutaraldehyde, orthophthalaldehyde, and formaldehyde destroy
Some sterilization and disinfection procedures are inappropriate for microorganisms and viruses by inactivating proteins and nucleic
certain types of material. acids. Chlorhexidine is a biguanide extensively used in antiseptic
products. Ethylene oxide is a gaseous sterilizing agent that destroys
microbes by reacting with proteins. Sodium hypochlorite (liquid
5.3 ■ Using Heat to Destroy Microorganisms
bleach) is one of the least expensive and most readily available forms
and Viruses
of chlorine. Chlorine dioxide is used as a sterilant and disinfec-
Moist Heat tant. Iodophores are iodine-releasing compounds used as antiseptics.
Moist heat destroys microorganisms by causing irreversible coagula- Metals interfere with protein function. Silver-containing compounds
tion of their proteins. Pasteurization utilizes a brief heat treatment to are used to prevent wound infections. Ozone is used as an alterna-
destroy spoilage and disease-causing organisms. Pressure cookers and tive to chlorine in the disinfection of drinking water and wastewater.
autoclaves use pressurized steam to achieve temperatures that can kill Peroxide and peracetic acid are both strong oxidizing agents that
endospores (figure  5.3). The most important aspect of the commercial can be used alone or in combination as sterilants. Phenolics destroy
canning process is to ensure that endospores of Clostridium botulinum cytoplasmic membranes and denature proteins; triclosan is used in
are destroyed. lotions and deodorant soaps. Quaternary ammonium compounds are
cationic detergents; they are non-toxic enough to be used to disinfect
Dry Heat food preparation surfaces.
Incineration burns cell components to ashes. Temperatures achieved
in hot air ovens destroy cell components and irreversibly denature 5.6 ■ Preservation of Perishable Products
proteins.
Chemical Preservatives
5.4 ■ Using Other Physical Methods Benzoic, sorbic, and propionic acids are sometimes added to foods
to Remove or Destroy Microbes to prevent microbial growth. Nitrate and nitrite are added to some
foods to inhibit the germination and subsequent growth of Clostridium
Filtration (figure 5.5)
botulinum endospores.
Membrane filters and depth filters retain microorganisms while let-
ting the suspending fluid pass through. High-efficiency particulate air Low-Temperature Storage
(HEPA) filters remove nearly all microorganisms from air. Low temperatures above freezing inhibit microbial growth. Freezing
essentially stops all microbial growth.
Radiation (figure 5.6)
Ionizing radiations destroys cells by damaging cell structures and pro- Reducing the Available Water
ducing reactive oxygen species. Ultraviolet light damages the structure Sugar and salt draw water out of cells, preventing the growth of micro-
and function of nucleic acids. Microwaves kill microorganisms by organisms. Lyophilization is used for preserving food. The food is first
generating heat. frozen and then dried in a vacuum.
124 Chapter 5 Control of Microbial Growth

Review Questions
Short Answer 8. Aseptically boxed juices and cream containers are processed using
1. How is preservation different from pasteurization? which of the following heating methods?
2. What is the most chemically resistant non-spore-forming bacterial a) Canning
pathogen? b) High-temperature–short-time (HTST) method
3. Explain why it takes longer to kill a population of 109 cells than it c) Low-temperature–long-time (LTLT) method
does to kill a population of 103 cells. d) Ultra-high-temperature (UHT) method
4. What is the primary reason that wine is pasteurized? 9. Commercial canning processes are designed to ensure destruction
5. What is the primary reason that milk is pasteurized? of which of the following?
6. When canning, why are low-acid foods processed at higher tem- a) All vegetative bacteria
peratures than high-acid foods? b) All viruses
7. How are heat-sensitive liquids sterilized? c) Endospores of Clostridium botulinum
8. How does microwaving a food product kill bacteria? d) E. coli
9. How is an iodophore different from a tincture of iodine? e) Mycobacterium tuberculosis
10. Name two products commonly sterilized using ethylene oxide gas. 10. Which of the following is false?
a) A high-level disinfectant cannot be used as a sterilant.
Multiple Choice
b) Critical items must be sterilized before use.
1. Unlike a disinfectant, an antiseptic
c) Low numbers of endospores may remain on semicritical items.
a) sanitizes objects rather than sterilizes them.
d) Standard sterilization procedures do not destroy prions.
b) destroys all microorganisms.
e) Quaternary ammonium compounds can be used to disinfect
c) is non-toxic enough to be used on human skin. food preparation surfaces.
d) requires heat to be effective.
e) can be used in food products. Applications
2. The D value is defined as the time it takes to kill 1. An agriculture extension agent is preparing pamphlets on prevent-
a) all bacteria in a population. ing the spread of disease. In the pamphlet, he must explain the
appropriate situations for using disinfectants around the house.
b) all pathogens in a population.
What situations should the agent discuss?
c) 99.9% of bacteria in a population.
2. As a microbiologist representing a food corporation, you have
d) 90% of bacteria in a population. been asked to serve on a health food panel to debate the need for
e) 10% of bacteria in a population. chemical preservatives in foods. Your role is to prepare a statement
3. Which of the following is the most resistant to destruction by that compares the benefits of chemical preservatives and the risks.
chemicals and heat? What points must you bring up that indicate the benefits of chemi-
a) Bacterial endospores cal preservatives?
b) Fungal spores
Critical Thinking +
c) Mycobacterium tuberculosis
1. This graph shows the time it takes to kill populations of the same
d) E. coli
microorganism under different conditions. What conditions would
e) HIV explain the differences in lines a, b, and c?
4. Ultraviolet light kills bacteria by
Number of surviving cells (logarithmic scale)

a) generating heat.
b) damaging DNA.
c) inhibiting protein synthesis.
d) damaging cell walls.
e) damaging cytoplasmic membranes.
a
5. Which concentration of alcohol is the most effective germicide?
b
a) 100% b) 75% c) 50% d) 25% e) 5%
6. Which of the following can most reliably be used as a sterilant?
a) Alcohol b) Phenolic compounds c) Ethylene oxide gas c

d) Iodine
7. All of the following are routinely used to preserve foods except
a) high concentrations of sugar. Time
b) high concentrations of salt.
c) benzoic acid. 2. This diagram shows the filter paper method used to evaluate the
d) freezing. inhibitory effect of chemical agents, heavy metals, and antibiot-
e) ethylene oxide. ics on bacterial growth. A culture of a test bacterium is spread
 Part I Life and Death of Microorganisms 125

uniformly over the surface of an agar plate. Small filter paper discs
containing the material to be tested are then placed on the surface Control disc
of the medium. A disc that has been soaked in sterile distilled water
E A Filter paper discs
is sometimes added as a control. After incubation, a lawn (film of soaked in material
growth) will cover the plate, but a clear zone will surround those to be tested (A–E)
discs that contain an inhibitory compound. The size of the zone Clear area D B
reflects several factors, one of which is the effectiveness of the around disc C Bacterial growth
inhibitory agent. What are two other factors that might affect the size (lawn) on agar
surface
of the zone of inhibition? What is the purpose of the control disc? If
a clear area were apparent around the control disc, how would you
interpret the observation?
6 Metabolism: Fueling Cell Growth
KEY TERMS
Adenosine Triphosphate (ATP)
The energy currency of cells.
Hydrolysis of the bonds between
its phosphate groups can be used
Photophosphorylation Synthesis
of ATP using the energy of a proton
motive force created by harvesting
radiant energy.
to power endergonic (energy- Precursor Metabolites Metabolic
consuming) reactions. intermediates that can be either
Anabolism Processes that used to make the subunits of
synthesize and assemble the subunits macromolecules or oxidized to
of macromolecules, using energy of generate ATP.
ATP; biosynthesis.
Proton Motive Force Form of
Catabolism Processes that energy generated as an electron
harvest energy released during the transport chain moves protons across
breakdown of compounds such as a membrane to create a chemiosmotic
glucose, using it to synthesize ATP. gradient.
Electron Transport Chain Group Respiration Metabolic process that
of membrane-embedded electron transfers electrons stripped from a
carriers that pass electrons from one chemical energy source to an electron
to another, and, in the process, create transport chain, generating a proton
a proton motive force. motive force that is then used to
Enzyme A protein that functions as synthesize ATP.
a catalyst, speeding up a biological Substrate-Level
Wine—a beverage produced using microbial metabolism. reaction. Phosphorylation Synthesis of
Fermentation Metabolic process ATP using the energy released in
that stops short of oxidizing glucose an exergonic (energy-releasing)
or other organic compounds chemical reaction during the
A Glimpse of History completely, using an organic breakdown of the energy source.
intermediate as a terminal electron Terminal Electron Acceptor
In the 1850s, Louis Pasteur tried to determine how alcohol develops from acceptor. Chemical that is ultimately reduced
grape juice. Biologists had already noticed that when grape juice is held as a consequence of fermentation or
Oxidative Phosphorylation
in large vats, alcohol and CO2 are produced and the number of yeast cells respiration.
Synthesis of ATP using the energy
increases. They concluded that multiplying cells were converting sugar in of a proton motive force created by
the juice to alcohol and CO2. Pasteur agreed, but could not convince two harvesting chemical energy.
very powerful and influential German chemists, Justus von Liebig and
Friedrich Wöhler, who refused to believe that microorganisms caused
the breakdown of sugar. Both men mocked the hypothesis, publishing
pictures of yeast cells looking like miniature animals taking in grape juice
through one end and releasing CO2 and alcohol through the other.

A
ll cells need to accomplish two fundamental tasks to
Pasteur studied the relationship between yeast and alcohol produc-
tion using a strategy commonly employed by scientists today—that is, grow. They must continually synthesize new parts—
simplifying the experimental system so that relationships can be more including cell walls, membranes, ribosomes, and nucleic
easily identified. First, he prepared a clear solution of sugar, ammonia, acids—that allow the cell to enlarge and eventually divide. In
mineral salts, and trace elements. He then added a few yeast cells. As addition, cells need to harvest energy and convert it to a form that
the yeast grew, the sugar level decreased and the alcohol level increased, can power the various energy-consuming reactions. The sum total
indicating that the sugar was being converted to alcohol as the cells of all chemical reactions in a cell is called metabolism.
multiplied. This strongly suggested that living cells caused the chemical Microbial metabolism is important to humans for a num-
transformation. Liebig, however, still would not believe that the process ber of reasons. For example, as scientists look for new supplies
was occurring inside microorganisms. To convince him, Pasteur tried to of energy, some are investigating biofuels—fuels made from
extract something from inside the yeast cells that would convert the sugar.
renewable biological sources such as plants and organic wastes.
He failed, like many others before him.
Metabolic pathways of microbes are used to produce biofuels,
In 1897, Eduard Buchner, a German chemist, showed that crushed
yeast cells could convert sugar to ethanol and CO2. We now know that the breaking down solid materials such as corn stalks, sugar cane, and
cells’ enzymes carried out this transformation. For these pioneering stud- wood to make ethanol. As another example, cheese-makers add
ies, Buchner was awarded a Nobel Prize in 1907. He was the first of many Lactococcus and Lactobacillus species to milk because the meta-
investigators who received Nobel Prizes for studies on the processes by bolic wastes of these bacteria contribute to the flavor and texture
which cells degrade sugars. of various cheeses. Microbial metabolism is also important in the
126
 Part I Life and Death of Microorganisms 127

laboratory, because products characteristic of specific microbes Metabolism can be separated into two components—catabolism
can be used as identifying markers. In addition, the metabolic and anabolism (figure  6.1). Catabolism is the set of processes
pathways of organisms such as E. coli serve as an invaluable that degrade compounds, releasing their energy. Cells capture that
model for studying similar pathways in eukaryotic cells, including energy and use it to make ATP (see figure  2.11). In contrast,
those of humans. Finally, metabolic processes unique to prokary- anabolism, or biosynthesis, is the set of processes that cells use
otes are potential targets for antimicrobial drugs. to synthesize and assemble the subunits of macromolecules, using
ATP for energy. The subunits of macromolecules include amino
6.1 ■ Principles of Metabolism acids, nucleotides, and lipids. ATP, p. 24
Although catabolism and anabolism are often discussed sepa-
Learning Outcomes rately, they are intimately linked. As mentioned, ATP made during
catabolism is used in anabolism. In addition, some of the compounds
1. Compare and contrast catabolism and anabolism.
produced during catabolism serve as precursor metabolites. These
2. Describe the energy sources used by photosynthetic organisms and
are molecules that can be diverted from the catabolic pathways and
chemoorganoheterotrophs.
used to make subunits such as amino acids.
3. Describe the components of metabolic pathways (enzymes, ATP,
chemical energy sources, redox reactions, and electron carriers)
and the role of precursor metabolites. Harvesting Energy
4. Describe the roles of the three central metabolic pathways.
Energy is the capacity to do work. It can exist as potential energy
5. Distinguish between respiration and fermentation.
(stored energy) and kinetic energy (energy of motion) (figure 6.2).
Potential energy can be stored in various forms including chemi-
CATABOLISM ANABOLISM cal bonds, a rock on a hill, or water behind a dam.
Energy in the universe can never be created or destroyed; how-
Energy source
(glucose) ever, it can be changed from one form to another. In other words,
although energy cannot be created, potential energy can be con-
verted to kinetic energy and vice versa, and one form of potential
Cell structures energy can be converted to another. Hydroelectric dams unleash the
(cell wall, membrane, potential energy of water stored behind a dam, creating the kinetic
ribosomes, surface energy of moving water. This can be captured to generate an electri-
structures)
cal current, which can then be used to charge a battery.
Energy Photosynthetic organisms harvest the energy of sunlight,
using it to power the synthesis of organic compounds from CO2.
Macromolecules By doing so, they convert the kinetic energy of photons (particles
(proteins, nucleic acids,
polysaccharides, lipids)

Energy
Subunits
(amino acids,
nucleotides, sugars,
fatty acids)

Energy
Precursor
metabolites

Waste products Nutrients

(acids, carbon (source of nitrogen,
dioxide) sulfur, etc.)

Catabolic processes harvest Anabolic processes

the energy released during the (biosynthesis) synthesize and
breakdown of compounds and assemble subunits of macro-
use it to make ATP. The molecules that make up the cell
processes also produce structures. The processes use
precursor metabolites used in the ATP and precursor metabo-
biosynthesis. lites produced in catabolism.

FIGURE 6.2 Forms of Energy Potential energy is stored energy,

FIGURE 6.1 The Relationship Between Catabolism such as water held behind a dam. Kinetic energy is the energy of
and Anabolism motion, such as movement of water from behind the dam.
? Which subunits make up proteins? Which make up nucleic acid? ? What is energy?
128 Chapter 6 Metabolism: Fueling Cell Growth

that travel at the speed of light) to the potential energy of chemical energy is released in the reaction. The reaction is said to be
bonds. Chemoorganotrophs obtain energy by degrading organic exergonic. In contrast, if the products have more free energy than
compounds. Thus, most chemoorganotrophs depend on the meta- the starting compounds, the reaction requires an input of energy
bolic activities of photosynthetic organisms (figure 6.3). and is termed endergonic.
The amount of energy released by breaking down a com- The change in free energy for a given reaction is the same
pound can be explained by the concept of free energy, the energy regardless of the number of steps involved. For example, convert-
available to do work. From a biological perspective, this is the ing glucose to CO2 and water in a single step releases the same
energy released when a chemical bond is broken. In a chemical amount of energy as degrading it in a series of steps. Cells use
reaction, some bonds are broken and others are formed. If the multiple steps when degrading compounds, harvesting the energy
starting compounds have more free energy than the products, released in an exergonic reaction by having it power an ender-
gonic one.

Components of Metabolic Pathways

The series of sequential chemical reactions that converts a starting
compound to an end product is called a metabolic pathway
(figure 6.4). Pathways can be linear, branched, or cyclical. Like
the flow of rivers controlled by dams, their activities can be
adjusted at certain points. This allows a cell to regulate certain
processes, ensuring that specific molecules are produced in pre-
cise quantities when needed. If a metabolic step is blocked, prod-
ucts “downstream” of that blockage will not be made.
The intermediates and end products of metabolic pathways
Radiant energy Photosynthetic organisms harvest the energy
(sunlight) of sunlight and use it to power the synthesis are sometimes organic acids, which are weak acids. Depending
of organic compounds from CO2. This on the pH of the environment, these occur as either the undissoci-
converts radiant energy to chemical energy. ated or dissociated (ionized) form. Biologists often use the names
of the two forms interchangeably—for example, pyruvic acid and
pyruvate (an ion). Note, however, that at the near-neutral pH
inside the cell, the ionized form predominates, whereas outside of
the cell, the acid often predominates. ion, p. 20 pH, p. 23
To understand what metabolic pathways accomplish, it is
important to be familiar with their critical components. These
include enzymes, ATP, the chemical energy source and terminal
electron acceptor, and electron carriers.

The Role of Enzymes

Enzymes are proteins that function as biological catalysts, accel-
erating the conversion of one substance, the substrate, into
another, the product. A specific enzyme facilitates each step of a
Chemical energy Chemoorganotrophs degrade organic
compounds, harvesting chemical energy.
metabolic pathway. Without enzymes, energy-yielding reactions
(organic compounds)
would still occur, but at rates so slow they would be insignificant.
 enzymes, p. 135  proteins, p. 25
An enzyme catalyzes a chemical reaction by lowering the
activation energy—the energy it takes to start a reaction
(figure 6.5). Even an exergonic reaction has an activation energy,
and by lowering this barrier, an enzyme speeds the reaction.
Enzymes will be described in more detail later in the chapter.

The Role of ATP

Adenosine triphosphate (ATP) is the main energy currency of
cells, serving as the ready and immediate donor of free energy.
The molecule is composed of ribose, adenine, and three phos-
FIGURE 6.3 Most Chemoorganotrophs Depend on phate groups arranged in tandem (see figure  2.11). Adenosine
Photosynthetic Organisms diphosphate (ADP) can be viewed as an acceptor of free energy.
? How do chemoorganotrophs require the activities of Cells produce “energy currency” by using energy to add an
photosynthetic organisms? inorganic phosphate group (Pi) to ADP, forming ATP. That energy
 Part I Life and Death of Microorganisms 129

Starting compound Intermediatea Intermediateb End product

(a) Linear metabolic path

pathway

Intermediateb1 End product1

Starting compound Intermediatea

(b) Branched metabolic p

pathway Intermediateb2 End product2

Starting compound

Intermediated

End product
Intermediatea
FIGURE 6.4 Metabolic Pathways Metabolic
pathways use a series of sequential chemical reactions to
convert starting compounds into intermediates and then,
Intermediatec ultimately, into end products. A metabolic pathway can be
(a) linear, (b) branched, or (c) cyclical.
Intermediateb ? In a branched metabolic pathway, how might a cell control
(c) Cyclical metabolic pathway which end product is made most often?

FIGURE 6.5 The Role of Enzymes Enzymes function as biological

catalysts. (a) An enzyme catalyzes a chemical reaction by lowering the
activation energy of the reaction. (b) A specific enzyme facilitates each
step of a metabolic pathway.
? What is activation energy?

Activation
Activation energy
energy without an
enzyme
Relative energy

Energy of with an
reactants enzyme

Energy of
products

Progress of reaction

(a)

Enzyme a Enzyme b Enzyme c

Starting compound Intermediatea Intermediateb End product

(b)
130 Chapter 6 Metabolism: Fueling Cell Growth

Unstable (high-energy) bonds Terminal

Energy electron
P~ P ~ P sources acceptors
ATP
Energy
Pi Pi released
H2 Organic Organic
carbon carbon
compounds compounds

Relative tendency to give up electrons

Relative tendency to give up electrons

Energy used H 2S CO2
The energy comes Energy released
from catabolic 0
The energy drives S SO4
reactions. anabolic reactions.
P~ P
FeOOH
ADP
Fe2+
FIGURE 6.6 ATP Cells produce ATP by using energy to add
inorganic phosphate (Pi) to ADP. Energy is released in the reverse
reaction, which breaks the phosphate bond to convert ATP to
ADP + Pi. NH4+
NO2– (to form NH4+)
? What characteristic of the structure of ATP makes its phosphate
NO3– (to form NH4+)
bonds “high energy”?
Mn2+
MnO2
currency can then be “spent” by removing the phosphate group,
releasing energy and again yielding ADP and Pi (figure 6.6). Cells
constantly produce ATP during exergonic reactions of catabolism NO3– (to form NH2)
and then use it to power endergonic reactions of anabolism. O2
Chemoorganotrophs use two different processes to make
ATP. In substrate-level phosphorylation, the energy released in (a) Energy is released when electrons are moved from an energy source with a
an exergonic reaction is used to power the addition of Pi to ADP; low affinity for electrons to a terminal electron acceptor with a higher affinity.
in oxidative phosphorylation the energy of a proton motive force
Glucose Terminal Inorganic Terminal
drives the reaction. Recall from chapter 3 that proton motive as an electron energy electron
force is the form of energy that results from the electrochemical energy source acceptors sources acceptors
gradient established by the electron transport chain (see fig-
ure  3.27). Details of this process will be discussed later in the
chapter. Photosynthetic organisms can generate ATP by
photophosphorylation. This uses the sun’s radiant energy and an Glucose H2
electron transport chain to create a proton motive force. The
Relative tendency to give up electrons

Relative tendency to give up electrons

Relative tendency to give up electrons

mechanism will be discussed later. oxidative phosphorylation, p. 142 H2S CO2
proton motive force, p. 55 photophosphorylation, p. 152 Pyruvate

The Role of the Chemical Energy Source

and the Terminal Electron Acceptor Fe2+
To understand how cells obtain energy, it is helpful to recall from
chapter 2 that certain atoms are more electronegative than others,
meaning they have a greater affinity (attraction) for electrons (see
table  2.2). Likewise, certain molecules have a greater affinity for
NO3–
electrons than other molecules, a characteristic that relates to the (to form
electronegativities of the atoms that make up the molecules. When NH4+)
electrons move from a molecule that has a relatively low electron
affinity (tends to give up electrons) to one that has a higher elec-
tron affinity (tends to accept electrons), energy is released. This is
how cells obtain the energy used to make ATP. They remove O2 O2
electrons from glucose or another low electron affinity chemical,
and donate them to a molecule such as O2 that has a higher affin- (b) Three examples of (c) Three examples of
ity. The chemical that serves as the electron donor is the energy chemoorganotrophic chemolithotrophic metabolism
source, and the one that ultimately accepts those electrons is the metabolism
terminal electron acceptor. The greater the difference between FIGURE 6.7 Chemical Energy Sources and Terminal Electron
the electron affinities of the energy source and the terminal elec- Acceptors
tron acceptor, the more energy released (figure 6.7). electrons, ? If Mn2+ is used as an energy source, which two molecules on the
p. 17 electronegativity, p. 21 chart in (a) could serve as terminal electron acceptors?
 Part I Life and Death of Microorganisms 131

Transfer of electrons The Role of Electron Carriers

e– When electrons are removed from the energy source, they are not
e– removed all at once, nor are they transferred directly to the termi-
Compound + Compound Compound X + Compound Y nal electron acceptor. Instead, the energy is harvested in a step-
X Y (oxidized) (reduced) wise process, with the electrons initially transferred to electron
carriers. Cells have several different types of electron carriers,
X loses electron(s).
Y gains electron(s). X is oxidized by the reaction.
each with distinct roles (table 6.1). The electron carriers are usu-
X is the reducing agent. Y is reduced by the reaction. ally referred to by abbreviations that represent their oxidized and
Y is the oxidizing agent. reduced forms, respectively:
■ NAD+/NADH (nicotinamide adenine dinucleotide)
FIGURE 6.8 Oxidation-Reduction Reactions The molecule that
loses one or more electrons is oxidized by the reaction; the one ■ NADP+/NADPH (nicotinamide adenine dinucleotide phosphate)
that gains those electrons is reduced.
■ FAD/FADH2 (flavin adenine dinucleotide)
? Why is hydrogenation a reduction?
These electron carriers can also be considered hydrogen carri-
ers because, along with electrons, they carry protons. FADH2
As a group, prokaryotes are remarkably diverse with respect carries two electrons and two protons; NADH and NADPH
to the energy sources and terminal electron acceptors they can use each carry two electrons and one proton. Note, however, that
(see figure 6.7a and b). E. coli and other chemoorganotrophs use the location of protons in biological reactions is often ignored
organic compounds such as glucose for an energy source, whereas because in aqueous solutions protons—unlike electrons—do
chemolithotrophs use hydrogen sulfide or another inorganic not require carriers.
chemical. Aerobic organisms can use O2 as a terminal electron Reduced electron carriers represent reducing power because
acceptor, whereas anaerobes use only molecules other than O2. they can easily transfer their electrons to another chemical that has
chemoorganotrophs p. 93 chemolithotrophs, p. 93 a higher affinity for electrons. By doing so, they raise the energy
Cells remove electrons from the energy source through a level of the recipient molecule. NADH and FADH2 transfer their
series of oxidation-reduction reactions, or redox reactions. The electrons to the electron transport chain, which then uses the
substance that loses electrons is oxidized by the reaction; the one energy to generate a proton motive force. In turn, this drives the
that gains those electrons is reduced (figure 6.8). When electrons synthesis of ATP in the process of oxidative phosphorylation.
are removed from a biological molecule, protons (H+) often fol- Ultimately the electrons are transferred to the terminal electron
low. The result is that an electron-proton pair, or hydrogen atom, acceptor. The electrons carried by NADPH have an entirely differ-
is removed. Thus, dehydrogenation (the removal of a hydrogen ent fate; they are used to reduce compounds during biosynthetic
atom) is an oxidation. Correspondingly, hydrogenation (the addi- reactions. Note, however, that many microbial cells can convert
tion of a hydrogen atom) is a reduction. reducing power in the form of NADH to NADPH.

TABLE 6.1 Electron Carriers

Oxidized Form Reduced Form Typical Fate of
Carrier (Accepts Electrons) (Donates Electrons) Electrons Carried

e–
Electron Electron
carrier
Nicotinamide adenine NAD+ + 2 e– + 2 H+  NADH + H+ Used to generate a
dinucleotide (carries proton motive force that
2 electrons and 1 proton) can drive ATP synthesis
Flavin adenine dinucleotide FAD+ + 2 e– + 2 H+  FADH2 Used to generate a
(carries 2 electrons and proton motive force that
2 protons; i.e., 2 hydrogen can drive ATP synthesis
atoms)
Nicotinamide adenine NADP+ + 2 e– + 2 H+  NADPH + H+ Biosynthesis
dinucleotide phosphate
(carries 2 electrons and
1 proton)
132 Chapter 6 Metabolism: Fueling Cell Growth

Precursor Metabolites Glucose molecules

Certain intermediates of catabolic pathways can be used in anabolic

pathways. These intermediates—precursor metabolites—serve
as carbon skeletons from which subunits of macromolecules can
be made (table 6.2). For example, the precursor metabolite pyru-
vate can be converted to any one of three amino acids: alanine,
leucine, or valine.
Recall from chapter 4 that E. coli can grow in glucose-salts
medium, which contains only glucose and a few inorganic salts.
This means that the glucose is serving two purposes in the cell: To: To:
(1) the energy source, and (2) the starting point from which all cell Lipid Amino acid
synthesis synthesis
components are made—including proteins, lipids, carbohydrates,
and nucleic acids. When E. coli cells degrade glucose molecules,
they use a series of steps that not only release energy but also form
a dozen or so precursor metabolites. Other organisms use the same
steps but sometimes lack certain biosynthetic capabilities. Any
essential compounds that a cell cannot synthesize must be pro-
To: To:
vided from an external source. glucose-salts medium, p. 95 Carbohydrate Nucleic acid
It is important to realize that the glucose molecules in E. coli synthesis synthesis
and other cells can have different fates. In essence, the catabolic
pathways are like an extensive highway system for millions of glu-
cose molecules (figure 6.9). Glucose molecules that remain on the
“highway” are oxidized completely to CO2, releasing the maximum
amount of energy. Some breakdown intermediates, however, can
“exit at an off ramp” to be used in biosynthesis. The “off ramps” are
located at the steps immediately after a precursor metabolite is made. CO2 molecules + energy
So once a precursor metabolite is made in catabolism, it can be fur- FIGURE 6.9 Metabolic Highway The millions of glucose
ther oxidized to release energy, or it can be used in biosynthesis. molecules that continually enter a cell can have different fates. Some
may be oxidized completely to release the maximum amount of
energy, and others will be used in biosynthesis.
Overview of Catabolism ? If precursor metabolites were shown on this diagram, where
Catabolism of glucose, the preferred energy source of many cells, would they be located?
encompasses two key processes: (1) oxidizing glucose molecules to
generate ATP, reducing power, and precursor metabolites; and Central Metabolic Pathways
(2) transferring the electrons carried by NADH and FADH2 (reducing Three key metabolic pathways—the central metabolic
power) to the terminal electron acceptor. The central metabolic path- pathways—gradually oxidize glucose to CO2 (figure 6.10). The
ways do the former, and respiration and fermentation do the latter. pathways are catabolic, but the precursor metabolites and reducing

TABLE 6.2 Precursor Metabolites

Precursor Metabolite Pathway Generated Biosynthetic Role

Glucose 6-phosphate Glycolysis Lipopolysaccharide

Fructose 6-phosphate Glycolysis Peptidoglycan
Dihydroxyacetone phosphate Glycolysis Lipids (glycerol component)
3-phosphoglycerate Glycolysis Proteins (the amino acids cysteine, glycine, and serine)
Phosphoenolpyruvate Glycolysis Proteins (the amino acids phenylalanine, tryptophan, and tyrosine)
Pyruvate Glycolysis Proteins (the amino acids alanine, leucine, and valine)
Ribose 5-phosphate Pentose phosphate cycle Nucleic acids and proteins (the amino acid histidine)
Erythrose 4-phosphate Pentose phosphate cycle Proteins (the amino acids phenylalanine, tryptophan, and tyrosine)
Acetyl-CoA Transition step Lipids (fatty acids)
α-ketoglutarate TCA cycle Proteins (the amino acids arginine, glutamate, glutamine, and proline)
Oxaloacetate TCA cycle Proteins (the amino acids aspartate, asparagine, isoleucine, lysine,
methionine, and threonine)

Some organisms use succinyl-CoA as a precursor in heme biosynthesis; E. coli uses glutamate.
 Part I Life and Death of Microorganisms 133

GLUCOSE

2 Pentose phosphate Yields

1 Glycolysis Reducing
pathway
Oxidizes glucose to pyruvate ~ ~ +
power
Starts the oxidation of glucose
ATP
by substrate-level
phosphorylation

Yields Reducing
power

Biosynthesis 5 Fermentation
Reduces pyruvate
or a derivative

Pyruvate Pyruvate Acids, alcohols, and gases

3a Transition step
CO2 CO2
Yields
Reducing
power

Acetyl- Acetyl-
CoA CoA

x2 CO2

CO2

3b TCA cycle
Incorporates an acetyl
group and releases CO2
(TCA cycles twice)

Yields
~ ~ + Reducing
power
ATP
by substrate-level 4 Respiration
phosphorylation Uses the electron transport
chain to convert reducing
power to proton motive force
FIGURE 6.10 Overview of Metabolism (1) Glycolysis, (2) the pentose phosphate
pathway, (3a) the transition step, and (3b) the tricarboxylic acid cycle (TCA cycle) are used to
gradually oxidize glucose completely to CO2. Together, these pathways produce ATP, reducing
power, and intermediates that function as precursor metabolites (depicted as colored ovals). Yields
~ ~
(4) Respiration uses the reducing power to generate a proton motive force that is then
harvested to make ATP by oxidative phosphorylation, ultimately passing the electrons to ATP
O2 or another terminal electron acceptor. (5) Fermentation stops short of oxidizing glucose by oxidative
completely and instead uses pyruvate or a derivative as a terminal electron acceptor. phosphorylation

? What is the terminal electron acceptor in aerobic respiration?

134 Chapter 6 Metabolism: Fueling Cell Growth

power they generate can also be diverted for use in biosynthesis. Also, when using anaerobic respiration, microbes employ a modi-
To reflect the dual role of these pathways, they are sometimes fied version of the TCA cycle. Organisms that use respiration,
called amphibolic pathways (amphi meaning “both kinds”). The either aerobic or anaerobic, are said to respire.
central metabolic pathways include:
Fermentation
■ Glycolysis. 1 This pathway splits glucose and gradually
oxidizes it to form two molecules of pyruvate. Glycolysis Cells that cannot respire are limited by their relative inability to
provides the cell with a small amount of energy in the form recycle reduced electron carriers. A cell has only a limited number
of ATP, some reducing power, and six precursor metabolites. of carrier molecules; if electrons are not removed from the
Note that some microbial cells use an alternative series of reduced carriers, none will be available to accept electrons. As a
reactions called the Entner-Doudoroff pathway. consequence, no more glucose molecules can be broken down.
Fermentation provides a solution to this problem. 5 In this pro-
■ Pentose phosphate pathway. 2 This also breaks down glu- cess, cells break down glucose through glycolysis only and then
cose, but its primary role in metabolism is the production of use pyruvate or a derivative as a terminal electron acceptor. By
compounds used in biosynthesis, including reducing power in transferring the electrons carried by NADH to pyruvate or a
the form of NADPH and two precursor metabolites. A prod- derivative, NAD+ is regenerated. Although fermentation does not
uct of the pathway feeds into glycolysis. involve the TCA cycle, organisms that ferment often use certain
■ Tricarboxylic acid cycle (TCA cycle). 3a As a prelude to key steps of it to generate specific precursor metabolites required
this cycle, a single reaction called the transition step converts for biosynthesis. Fermentation oxidizes glucose only partially and,
the pyruvate from glycolysis into acetyl-CoA. 3b The TCA compared with respiration, produces relatively little ATP.
cycle then accepts the 2-carbon acetyl group, ultimately oxi- Table 6.3 summarizes the difference between aerobic respiration,
dizing it to release two molecules of CO2. The transition step anaerobic respiration, and fermentation.
and the TCA cycle together generate the most reducing power
of all the central metabolic pathways. They also produce three
MicroAssessment 6.1
precursor metabolites and ATP.
Catabolic pathways gradually oxidize an energy source to harvest
During the oxidation of glucose, a relatively small amount of ATP energy. A specific enzyme catalyzes each step. Substrate-level
is made by substrate-level phosphorylation. The reducing power phosphorylation uses exergonic chemical reactions to synthesize
accumulated during the oxidation steps, however, can be used to ATP; oxidative phosphorylation employs a proton motive force to
generate ATP by oxidative phosphorylation. do the same. Reducing power in the form of NADH and FADH2
is used to generate a proton motive force; the reducing power
of NADPH is utilized in biosynthesis. Precursor metabolites are
Respiration metabolic intermediates that can be used in biosynthesis. The
4 Respiration transfers the electrons extracted from glucose to central metabolic pathways generate ATP, reducing power, and
the electron transport chain. The electron transport chain then uses precursor metabolites.
the electrons to generate a proton motive force, a form of energy 1. How does the fate of electrons carried by NADPH differ from the
that can be harvested to make ATP by oxidative phosphorylation. fate of electrons carried by NADH?
In aerobic respiration, O2 serves as the terminal electron accep- 2. Why are the central metabolic pathways called amphibolic?
tor. Anaerobic respiration is similar to aerobic respiration, but it 3. Why does fermentation supply less energy than respiration? +
uses a molecule other than O2 as a terminal electron acceptor.

TABLE 6.3 ATP-Generating Processes of Prokaryotic Chemoorganoheterotrophs

ATP Generated by ATP Generated
Uses an Substrate-Level by Oxidative Total ATP
Electron Terminal Phosphorylation Phosphorylation Generated
Metabolic Transport Electron (Theoretical (Theoretical (Theoretical
Process Chain Acceptor Maximum) Maximum) Maximum)

Aerobic Yes O2 2 in glycolysis (net) 34 38

respiration 2 in the TCA cycle
4 total
Anaerobic Yes Molecule other than Number varies; however, the ATP yield of anaerobic respiration is less than that
respiration O2 such as nitrate of aerobic respiration but more than that of fermentation.
(NO3–), nitrite (NO2–),
sulfate (SO42–)
Fermentation No Organic molecule 2 in glycolysis (net) 0 2
(pyruvate or a 2 total
derivative)
 Part I Life and Death of Microorganisms 135

6.2 ■ Enzymes Mechanisms and Consequences

of Enzyme Action
Learning Outcomes
An enzyme has on its surface an active site, typically a rela-
6. Describe the active site of an enzyme, and explain how it relates
tively small crevice (figure  6.11). This is the critical site to
to the enzyme-substrate complex.
which a substrate binds by weak forces. The binding of the
7. Compare and contrast cofactors and coenzymes.
substrate to the active site causes the shape of the flexible
8. List two environmental factors that influence enzyme activity. enzyme to change slightly. This mutual interaction, or induced
9. Describe allosteric regulation. fit, results in a temporary intermediate called an enzyme-
10. Compare and contrast competitive enzyme inhibition and non- substrate complex. The substrate is held within this complex
competitive enzyme inhibition. in a specific orientation so that existing bonds are destabilized
and new ones can easily form, lowering the activation energy
Recall that enzymes are biological catalysts, which increase the of the reaction. The products are then released, leaving the
rate at which substrates are converted into products (see fig- enzyme unchanged and free to combine with new substrate
ure  6.5). They do this with extraordinary specificity and speed, molecules. Note that enzymes can also catalyze reactions that
usually acting on only one or a few substrates. They are neither join two substrates to create one product. Theoretically, all
consumed nor permanently changed during a reaction, allowing a enzyme-catalyzed reactions are reversible, but the free energy
single enzyme molecule to be rapidly used again and again. More change of certain reactions makes them effectively non-
than a thousand different enzymes exist in a cell. enzymes, p. 135 reversible.
The name of an enzyme usually reflects its function and ends The interaction of an enzyme with its substrate is very spe-
with the suffix -ase. For example, isocitrate dehydrogenase cific. The substrate fits into the active site like a hand into a glove.
removes a hydrogen atom (a proton-electron pair) from isocitrate. Not only must it fit spatially, but appropriate chemical interactions
Some groups of enzymes are referred to by their general such as hydrogen and ionic bonding need to occur to induce the
function—for example, proteases degrade proteins. fit. This requirement for a precise fit and interaction explains why,
with few exceptions, a unique enzyme is required to catalyze each
MicroByte reaction in a cell. Very few molecules of any particular enzyme
In only one second, the fastest enzymes can transform more than 104
are needed, however, as each can be used repeatedly.
substrate molecules into products.
hydrogen bonds, p. 21 ionic bonds, p. 20

Enzyme-substrate Products released

Substrate
complex formed

Enzyme

Active site

Enzyme
unchanged
(a)

Substrate Substrate
Enzyme
Enzyme

(b) (c)

FIGURE 6.11 Mechanism of Enzyme Action (a) The substrate binds to the active site, forming an enzyme-substrate complex. The
products are then released, leaving the enzyme unchanged and free to combine with new substrate molecules. (b) A model showing an enzyme
and its substrate. (c) The binding of the substrate to the active site causes the shape of the flexible enzyme to change slightly.
? Why are enzymes so specific with respect to the reactions they catalyze?
136 Chapter 6 Metabolism: Fueling Cell Growth

with them. The same coenzyme can assist different enzymes.

Because of this, far fewer different coenzymes are required than
enzymes. Like enzymes, coenzymes are recycled as they function
and, consequently, are needed only in very small quantities.
Most coenzymes are derived from vitamins (table 6.4). Many
prokaryotes can synthesize these vitamins and convert them to the
Enzyme necessary coenzymes. Humans and other animals, however, can-
not synthesize most vitamins, so the vitamins usually must be
ingested as part of the diet. In some cases, vitamins made by bac-
teria residing in the intestine can be absorbed. If an animal lacks a
vitamin, the functions of all the different enzymes whose activity
Cofactor requires the corresponding coenzyme are impaired. Thus, a single
vitamin deficiency has serious consequences.

Substrate Environmental Factors That

Influence Enzyme Activity
FIGURE 6.12 Some Enzymes Act in Conjunction with a Several environmental factors influence how well enzymes func-
Cofactor Cofactors are non-protein components, either coenzymes tion and in this way determine how rapidly microbes multiply
or trace elements.
(figure  6.13). Each enzyme has a narrow range of conditions—
? What is the function of the coenzyme FAD? including temperature, pH, and salt concentration—within which
it operates best. A 10°C rise in temperature approximately doubles
the speed of enzymatic reactions, until optimal activity is reached;
this explains why bacteria tend to grow more rapidly at higher
Cofactors temperatures. If the temperature is too high, however, proteins
Some enzymes act with the assistance of a non-protein component will denature and no longer function. Most enzymes operate best
called a cofactor (figure  6.12). Magnesium, zinc, copper, and at low salt concentrations and at pH values slightly above 7. Not
other trace elements often function as cofactors. Coenzymes are surprisingly, most microbes grow fastest under these same condi-
organic cofactors that function as loosely bound carriers of mole- tions. Some prokaryotes, however, particularly certain members
cules or electrons (table  6.4). They include the electron carriers of the Archaea, thrive in environments where conditions are
FAD, NAD+, and NADP+. trace elements, p. 93 extreme. They may require high salt concentrations, very acidic
All coenzymes transfer substances from one compound to conditions, or temperatures near boiling. Enzymes from these
another, but they function in different ways. Some remain bound organisms can be very important commercially because they func-
to the enzyme during the transfer process, whereas others separate tion in harsh conditions that are often typical of industrial settings.
from the enzyme, carrying the substance being transferred along temperature and growth requirements, p. 89 denaturation, p. 29

TABLE 6.4 Some Coenzymes and Their Function

Vitamin from Which
Coenzyme It Is Derived Substance Transferred Example of Use

Nicotinamide adenine Niacin Hydride ions (2 electrons and Carrier of reducing power
dinucleotide (NAD+) 1 proton)
Flavin adenine dinucleotide (FAD) Riboflavin Hydrogen atoms (2 electrons Carrier of reducing power
and 2 protons)
Coenzyme A Pantothenic acid Acyl groups Carries the acetyl group that
enters the TCA cycle
Thiamin pyrophosphate Thiamine Aldehydes Helps remove CO2 from pyruvate
in the transition step
Pyridoxal phosphate Pyridoxine Amino groups Transfers amino groups in amino
acid synthesis
Tetrahydrofolate Folic acid 1-carbon molecules 1-carbon donor in nucleotide
synthesis
 Part I Life and Death of Microorganisms 137

FIGURE 6.13 Environmental

Factors That Influence Enzyme
Optimum Activity (a) A rise in temperature
temperature increases the speed of enzymatic
Optimum pH
activity until the optimum

Enzyme activity
temperature is reached. If the
Enzyme activity

temperature gets too high, the

enzyme denatures and no longer
functions. (b) Most enzymes function
best at pH values slightly above 7.
? Why would an enzyme no longer
function once it denatures?

1 2 3 4 5 6 7 8 9 10 11 12 13
Temperature Acidic Basic

(a) (b)

Allosteric Regulation Allosteric enzymes generally catalyze the first step of a path-
Cells can rapidly adjust the activity of certain key enzymes, using way. If the pathway is biosynthetic, the end product generally acts
other molecules that reversibly bind to and distort them as the allosteric inhibitor—a mechanism called feedback
(figure 6.14). This is how cells regulate the activity of metabolic inhibition (figure 6.14c). This allows the cell to shut down a path-
pathways. The enzymes that can be controlled are allosteric (allo way when the product begins accumulating. For example, the
means “other” and stereos means “shape”). They have an alloste- amino acid isoleucine is an allosteric inhibitor of the first enzyme
ric site as well as an active site. When a regulatory molecule binds of the pathway that converts threonine to isoleucine. When the
to the allosteric site, the shape of the enzyme changes. This distor- level of isoleucine is relatively high, the pathway is shut down.
tion alters the relative affinity (chemical attraction) of the enzyme The binding of the isoleucine is reversible, however, so the
for its substrate. In some cases the binding of the regulatory mol- enzyme becomes active again if isoleucine levels decrease. Cells
ecule enhances the affinity for the substrate; in other cases it can also control the amount of enzyme they synthesize, a topic
decreases it. discussed in chapter 7. bacterial gene regulation, p. 179

Enzyme Enzyme
Allosteric
inhibitor

Substrate

Allosteric site Active site

(a) (b)

Allosteric inhibitor

Enzyme a Enzyme b Enzyme c

Starting compound Intermediatea Intermediateb End product

(c)

FIGURE 6.14 Regulation of Allosteric Enzymes (a) Allosteric enzymes have, in addition to the active site, an allosteric site. (b) The
binding of a regulatory molecule to the allosteric site causes the shape of the enzyme to change, altering the relative affinity of the enzyme for
its substrate. (c) The end product of a given biosynthetic pathway generally acts as an allosteric inhibitor of the first enzyme of that pathway.
? Why would a cell need to regulate enzyme activity?
138 Chapter 6 Metabolism: Fueling Cell Growth

TABLE 6.5 Characteristics of Enzyme Inhibitors

Type Characteristics

Competitive inhibition Inhibitor binds to the active site of the enzyme, blocking access of the substrate to that site. Competitive
inhibitors such as sulfa drugs are used as antibacterial medications.
Non-competitive inhibition Inhibitor changes the shape of the enzyme, so that the substrate can no longer bind the active site. This is a
(by regulatory molecules) reversible action that provides cells with a means to control the activity of allosteric enzymes.
Non-competitive inhibition Inhibitor permanently changes the shape of the enzyme, making the enzyme non-functional. Enzyme
(by enzyme poisons) poisons such as mercury are used in certain antimicrobial compounds.

Compounds that reflect a cell’s relative energy supply often vitamin folic acid. The drug does not affect human metabolism
regulate allosteric enzymes of catabolic pathways. This allows because humans cannot synthesize folic acid; it must be consumed
cells to adjust the flow of metabolites through these pathways in in foods or nutritional supplements. Sulfa drugs have a structure
response to changing energy needs. High levels of ATP inhibit similar to para-aminobenzoic acid (PABA), an intermediate in the
certain enzymes and, as a consequence, slow down catabolic pro- bacterial pathway for folic acid synthesis. Because of this, they fit
cesses. In contrast, high levels of ADP warn that a cell’s energy into the active site of the enzyme that normally uses PABA as a
stores are low and stimulate the activity of some enzymes. substrate. By doing so, they prevent the enzyme from binding
PABA. The greater the number of sulfa molecules relative to
PABA molecules, the more likely the active site of the enzyme
Enzyme Inhibition will be occupied by a sulfa molecule. Once the sulfa is removed,
Enzymes can be inhibited by a variety of compounds other than however, the enzyme functions normally with PABA as the sub-
the regulatory molecules just described (table 6.5). These inhibi- strate. sulfa drugs, p. 467
tory compounds can prevent microbial growth, so they are medi-
cally and commercially valuable. The site on the enzyme to which Non-Competitive Inhibition
an inhibitor binds determines whether it functions as a competitive Non-competitive inhibition occurs when the inhibitor binds to a
or non-competitive inhibitor. site other than the active site. The binding changes the shape of
the enzyme so that the substrate can no longer bind the active site.
Competitive Inhibition The allosteric inhibitors discussed earlier are non-competitive
In competitive inhibition, the inhibitor binds to the active site of inhibitors that have a reversible action. The effect of other non-
the enzyme, blocking access of the substrate to that site competitive inhibitors is permanent. For example, mercury inhibits
(figure  6.15). Generally the inhibitor has a chemical structure growth because it oxidizes the S–H groups of the amino acid cys-
similar to the normal substrate. teine in proteins. This converts cysteine to cystine, which cannot
A good example of competitive inhibition is the action of the form the important covalent disulfide bond (S–S). As a result, the
group of antimicrobial medications called sulfa drugs. These enzyme shape changes, making it non-functional; the inhibitor “poi-
inhibit an enzyme in the pathway bacteria use to synthesize the sons” the enzyme. cysteine, p. 26 disulfide bond, p. 27

Structural
PABA differences
(substrate)
H H
HO O N

C O S O

Sulfa
(inhibitor)

N N
H H H H

PABA Sulfanilamide
Enzyme
(a) (b)
FIGURE 6.15 Competitive Inhibition of Enzymes (a) The inhibitor competes with the normal substrate for binding to the active site. The
greater the proportion of inhibitor relative to substrate, the more likely the active site of the enzyme will be occupied by an inhibitor. (b) A
competitive inhibitor generally has a chemical structure similar to the normal substrate.
? Will the enzyme function if sulfa is removed?
 Part I Life and Death of Microorganisms 139

MicroAssessment 6.2 Comparison of the Central

TABLE 6.6
Enzymes facilitate the conversion of a substrate into a product with Metabolic Pathways
extraordinary speed and specificity. They are neither consumed nor Pathway Characteristics
permanently changed in the reaction. The activity of some enzymes
requires a cofactor. Environmental factors influence enzyme activity Glycolysis Glycolysis generates:
and, by doing so, determine how rapidly microorganisms multiply.
• 2 ATP (net) by substrate-level phosphorylation
The activity of allosteric enzymes can be regulated. A variety of
different compounds and conditions affect enzyme activity. • 2 NADH + 2 H+
4. Explain why sulfa drugs prevent bacterial growth without • six different precursor metabolites
harming the human host. Pentose The pentose phosphate cycle generates:
5. Explain the function of a coenzyme. phosphate • NADPH + H+ (amount varies)
cycle
6. Why is it important for a cell that allosteric inhibition • two different precursor metabolites
be reversible? +
Transition The transition step, repeated twice to oxidize
step two molecules of pyruvate to acetyl-CoA,
generates:
6.3 ■ The Central Metabolic • 2 NADH + 2 H+
Pathways • one precursor metabolite
TCA cycle The TCA cycle, repeated twice to incorporate
Learning Outcomes two acetyl groups, generates:
11. Make a simple diagram that integrates the central metabolic
• 2 ATP by substrate-level phosphorylation (may
pathways, including their starting and end products. involve conversion of GTP)
12. Compare and contrast each of the central metabolic pathways
• 6 NADH + 6 H+
with respect to the yield of ATP, reducing power, and number of
different precursor metabolites. • 2 FADH2
• two different precursor metabolites
The three central metabolic pathways—glycolysis, the pentose
phosphate pathway, and the tricarboxylic acid cycle—modify
organic molecules in a step-wise fashion, generating: The pathway can be viewed as having two phases (figure 6.16):
■ ATP by substrate-level phosphorylation ■ Investment or preparatory phase. 1 through 5 This
■ Reducing power in the form of NADH, FADH2, and NADPH consumes energy because two different steps each transfer
a high-energy phosphate group to the 6-carbon sugar. In
■ Precursor metabolites (see table 6.2) eukaryotic cells, both phosphates come from ATP, as shown
This section describes how a molecule of glucose is broken in figure 6.16. In bacteria, the first one is added as glucose is
down in the central metabolic pathways. Bear in mind, however, transported into the cell via group translocation; the next one
that many millions of molecules of glucose enter a cell, and differ- comes from ATP. The 6-carbon sugar is then split to yield
ent molecules can have different fates (see figure 6.9). For exam- two 3-carbon molecules, each with a phosphate molecule.
ple, a cell might oxidize one glucose molecule completely to CO2, group translocation, p. 56
thereby producing the maximum amount of ATP. Another glucose ■ Pay-off phase. 6 through 10 This oxidizes and rearranges
molecule might enter glycolysis, or perhaps the pentose phosphate the 3-carbon molecules, generating 1 NADH and 2 ATP in
pathway, only to be siphoned off as a precursor metabolite for use the process, and ultimately forms pyruvate. Note that the
in biosynthesis. The step and rate at which the various intermedi- steps of this phase occur twice for each molecule of glucose
ates are removed for biosynthesis will dramatically affect the that entered glycolysis. This is because the 6-carbon sugar
overall energy gain of catabolism. This is generally overlooked in was split into two 3-carbon molecules in the previous phase.
descriptions of the ATP-generating functions of these pathways A total of 2 NADH and 4 ATP are made from 1 molecule of
for the sake of simplicity. However, because these pathways serve glucose.
more than one function, the energy yields are only theoretical.
The pathways of central metabolism are compared in Yield of Glycolysis
table  6.6. The entire pathways with chemical formulas and For every glucose molecule degraded, the steps of glycolysis produce:
enzyme names are illustrated in Appendix IV.
■ ATP: 2 molecules of ATP, net gain (4 ATP molecules are
Glycolysis made in the pay-off phase, minus the 2 spent in the invest-
ment phase).
Glycolysis is the primary pathway used by many organisms to
convert glucose to pyruvate (figure 6.16). For each molecule of ■ Reducing power: The payoff phase converts 2 NAD+ to
glucose that enters, two molecules of pyruvate can be made. This 2 NADH + 2 H+.
generates a net gain of two molecules of ATP and two molecules ■ Precursor metabolites: Five intermediates of glycolysis as
of NADH. In addition, the pathway produces six different precur- well as the end product, pyruvate, are precursor metabolites
sor molecules needed by E. coli (see table 6.2). used by E. coli.
140 Chapter 6 Metabolism: Fueling Cell Growth

Pentose phosphate
GLUCOSE

Glycolysis Yields
FIGURE 6.16 Glycolysis This pathway oxidizes glucose to pyruvate,
2 Reducing

generating ATP by substrate-level phosphorylation, reducing power in the

pathway 1
Oxidizes glucose to pyruvate ~ ~ +
power
Starts the oxidation of glucose
ATP
by substrate-level
phosphorylation

form of NADH, and six different precursor metabolites.

? How many ATPs—total and net—are produced from breaking down one
Yields Reducing
power molecule of glucose in glycolysis?
Biosynthesis Fermentation
5
Reduces pyruvate
or a derivative

3a Transition step
Pyruvate Pyruvate Acids, alcohols, and gases
Glucose
CO2 CO2
Yields
Reducing
power

Acetyl- Acetyl-
CoA CoA

x2 CO2
ATP ~ ~ 1 ATP is expended to add a phosphate group.
CO2

~
TCA cycle

ADP
3b
Incorporates an acetyl
group and releases CO2
(TCA cycles twice)

Yields
~ ~ + Reducing
power
ATP
by substrate-level 4 Respiration
phosphorylation Uses the electron transport
chain to convert reducing
power to proton motive force

Yields
~ ~
Glucose
ATP
by oxidative
phosphorylation
6-phosphate

2 A chemical rearrangement occurs.

Fructose
6-phosphate

ATP ~ ~ 3 ATP is expended to add a phosphate group.

ADP ~

Fructose
1,6-bisphosphate
4 The 6-carbon molecule is split into two 3-carbon
molecules.

Dihydroxyacetone
phosphate
5 A chemical rearrangement of one of the
molecules occurs.
Glyceraldehyde
3-phosphate

NAD+ NAD+
6 The addition of a phosphate
NADH + H+ NADH + H+ group is coupled to a redox
reaction, generating NADH and
1,3-bisphospho- a high-energy phosphate bond.
glycerate ~ ~

ADP ~ ~ 7 ATP is produced by

substrate-level
ATP ~ ~ ~ ~ phosphorylation.

3-phospho-
glycerate

8 A chemical rearrangement occurs.

2-phospho-
glycerate

9 Water is removed, causing the

H2O H2O phosphate bond to become
Phospho- high-energy.
~

enolpyruvate

ADP ~ ~ 10 ATP is produced by

substrate-level
ATP ~ ~ ~ ~ phosphorylation.

Pyruvate
 Part I Life and Death of Microorganisms 141

Pentose Phosphate Pathway Yield of the Pentose Phosphate Pathway

The other central metabolic pathway used by cells to break down The yield of the pentose phosphate pathway varies, depending upon
glucose is the pentose phosphate pathway. This pathway is par- which of several possible alternatives are taken. It can produce:
ticularly important because of its contribution to biosynthesis. It ■ Reducing power: A variable amount of reducing power in
generates reducing power in the form of NADPH, and two of its the form of NADPH is produced.
intermediates—ribose 5-phosphate and erythrose 4-phosphate— ■ Precursor metabolites: Two intermediates of the pentose
are important precursor metabolites. A product of this complex phosphate pathway are precursor metabolites.
pathway is glyceraldehyde 3-phosphate (G3P), which can enter a
step in glycolysis for further breakdown. Transition Step
GLUCOSE
The transition step, which links the previous pathways to the
2
Pentose phosphate
pathway
Starts the oxidation of glucose
1
Glycolysis
Oxidizes glucose to pyruvate
Yields
~ ~ + Reducing
power
TCA cycle, involves several reactions catalyzed by a large
ATP
by substrate-level
phosphorylation

Pyruvate
multi-enzyme complex (figure  6.17). CO2 is first removed

Yields Reducing
power CO2
Biosynthesis
5 Fermentation
Reduces pyruvate
Transition step:
or a derivative

Pyruvate Pyruvate Acids, alcohols, and gases NAD+ CO2 is removed, a redox reaction generates
3a Transition step

Yields
CO2 CO2 CoA NADH, and coenzyme A is added.
Reducing
power

Acetyl- Acetyl-
CoA CoA

NADH + H+
x2 CO2

3b TCA cycle
CO2
CoA
Incorporates an acetyl
group and releases CO2
(TCA cycles twice)
Acetyl-CoA
Yields
~ ~ + Reducing
power
1 The acetyl group is transferred
ATP
by substrate-level 4 Respiration
CoA
to oxaloacetate to start a new
phosphorylation Uses the electron transport
chain to convert reducing
power to proton motive force

Yields
~ ~
round of the cycle.
ATP
by oxidative
phosphorylation

Oxaloacetate
NADH + H+ 2 A chemical
Citrate rearrangement occurs.
8 A redox reaction
generates NADH.

NAD+
Isocitrate
NAD+
3 A redox reaction
generates NADH
Malate and CO2 is
7 Water is added. removed.

NADH + H+
H2O
CO2

Fumarate
α-ketoglutarate
NAD+
4 A redox reaction
FADH2 generates NADH,
CoA
CO2 is removed,
and coenzyme A
6 A redox reaction NADH + H+ is added.
generates FADH2.
CO2
FAD CoA
Succinate Succinyl-CoA

5 The energy released

during CoA removal is CoA
C
~ ~ ~ + Pi
harvested to produce ATP.
ATP ADP

FIGURE 6.17 The Transition Step and the Tricarboxylic Acid Cycle The transition step links glycolysis and the TCA cycle, converting pyruvate
to acetyl-CoA; it generates reducing power and one precursor metabolite. The TCA cycle incorporates the acetyl group of acetyl-CoA and, using a series
of steps, releases CO2; it generates ATP, reducing power in the form of both NADH and FADH2, and two different precursor metabolites.
? Which generates more reducing power—glycolysis or the TCA cycle?
142 Chapter 6 Metabolism: Fueling Cell Growth

from pyruvate, a step called decarboxylation. Then, a redox reaction ■ Precursor metabolites: Two intermediates of the TCA
transfers electrons to NAD+, reducing it to NADH + H+. Finally, the cycle, formed in steps 3 and 8, are precursor metabolites.
remaining 2-carbon acetyl group is joined to coenzyme A to form
acetyl-CoA. MicroAssessment 6.3
In prokaryotic cells, all the central metabolic pathways occur
Glycolysis oxidizes glucose to pyruvate, yielding some ATP and
in the cytoplasm. In eukaryotic cells, however, the enzymes of NADH and six different precursor metabolites. The pentose phosphate
glycolysis and the pentose phosphate pathways are located in the pathway also oxidizes glucose, but more importantly, it produces two
cytoplasm, whereas those of the transition step and TCA cycle are different precursor metabolites and NADPH for biosynthesis. The
within the mitochondrial matrix. Because of this, eukaryotic cells transition step and the TCA cycle, each repeated twice, complete the
must transport pyruvate molecules into mitochondria for the tran- oxidation of glucose, yielding some ATP, a great deal of reducing
sition step to occur. cytoplasm, p. 52 mitochondria, p. 74 power, and three different precursor metabolites.
7. How does the “investment phase” of glycolysis effect the net
Yield of the Transition Step yield of ATP in that pathway?
The transition step occurs twice for every molecule of glucose 8. Which central metabolic pathway generates the most reducing
that enters glycolysis, oxidizing pyruvate to form acetyl-CoA. power?
Together, these generate: 9. Which compound contains more free energy—glucose or
oxaloacetate? On what did you base your conclusion? +
■ Reducing power: 2 NADH + 2 H+.
■ Precursor metabolites: One precursor metabolite (acetyl-
CoA).
6.4 ■ Respiration
Tricarboxylic Acid (TCA) Cycle Learning Outcomes
The tricarboxylic acid (TCA) cycle completes the oxidation of glu- 13. Describe the components of the electron transport chain and how
cose (see figure  6.17). The cycle incorporates the acetyl groups they generate a proton motive force.
from the transition step, ultimately releasing two molecules of CO2. 14. Compare and contrast the electron transport chains of eukaryotes
In addition to generating ATP and reducing power, the steps of the and prokaryotes.
TCA cycle form two more precursor metabolites (see table 6.2). 15. Describe how proton motive force is used to synthesize ATP and
1 The TCA cycle begins when CoA transfers its acetyl how the ATP yield of aerobic respiration is calculated.
group to the 4-carbon compound oxaloacetate, forming the 6-car-
bon compound citrate. 2 Citrate is then chemically rearranged to Respiration uses the reducing power generated in glycolysis, the
make isocitrate. 3 This is oxidized and a molecule of CO2 transition step, and the TCA cycle to synthesize ATP. The mecha-
removed, producing the 5-carbon compound α-ketoglutarate. nism, oxidative phosphorylation, involves two sequential pro-
During the oxidation, NAD+ is reduced to NADH + H+. 4 Then, cesses. First, the electron transport chain generates a proton
in a complex step, α-ketoglutarate is oxidized, CO2 is removed, motive force. Then, the enzyme ATP synthase uses the energy of
and CoA is added, producing the 4-carbon compound succinyl- the proton motive force to drive the synthesis of ATP.
CoA. During the process, NAD+ is reduced to NADH + H+. The remarkable process that links the electron transport chain
5 The CoA is then removed from succinyl-CoA, and the energy to ATP synthesis was proposed by the British scientist Peter
released is used to produce ATP by substrate-level phosphoryla- Mitchell in 1961. His hypothesis, now called the chemiosmotic
tion. Note that some bacteria and other cells make guanosine tri- theory, was widely dismissed initially. Only through years of self-
phosphate (GTP) rather than ATP at this step; GTP can be funded research was he able to convince others of its validity; he
converted to ATP. 6 Succinate is then oxidized to fumarate, as received a Nobel Prize in 1978.
FAD is reduced to FADH2. 7 A molecule of water is added to
fumarate, producing malate. 8 This compound is then oxidized
to oxaloacetate; note that oxaloacetate is the starting compound to
The Electron Transport Chain—
which acetyl-CoA is added to initiate the cycle. During this last Generating Proton Motive Force
step, NAD+ is reduced to NADH + H+. The electron transport chain is a group of membrane-embedded
electron carriers that pass electrons sequentially from one to
Yield of the TCA Cycle another, ejecting protons in the process (see figure 3.27). In pro-
The tricarboxylic acid cycle “turns” once for each acetyl-CoA that karyotes, the electron transport chain is located in the cytoplasmic
enters. Because two molecules of acetyl-CoA are generated for membrane, whereas in eukaryotic cells it is in the inner membrane
each glucose molecule that enters glycolysis, the breakdown of of mitochondria. cytoplasmic membrane, p. 52
one molecule of glucose causes the TCA cycle to turn twice. Because of the order of the different carriers in the electron
Together, these two turns generate: transport chain and their relative affinities for electrons, energy is
gradually released as the electrons are passed from one carrier to
■ ATP: 2 ATP produced in step 5. another, much like a ball falling down a flight of stairs
■ Reducing power: Redox reactions at steps 3, 4, 6, and 8 (figure  6.18). The energy release is coupled to the ejection of
produce a total of 6 NADH + 6 H+ and 2 FADH2. protons, moving them from the inside of the cell to the outside or,
 Part I Life and Death of Microorganisms 143

Electrons from the Flavoproteins are proteins to which a flavin is attached. FAD
energy source 2 e– and other flavins are synthesized from the vitamin riboflavin (see
Energy released is table 6.4).
used to generate a
proton motive force. General Mechanisms of Proton Ejection
An important characteristic of the electron carriers is that some
High energy
accept only hydrogen atoms (proton-electron pairs), whereas others
accept only electrons. The spatial arrangement of these two types of
carriers in the membrane causes protons to be shuttled from one side
of the membrane to the other. This occurs because a hydrogen car-
rier receiving electrons from an electron carrier must pick up pro-
tons, which come from inside the cell (or matrix of the mitochondrion)
due to the hydrogen carrier’s relative location in the membrane.
Electrons donated
Low energy to the terminal
Conversely, when a hydrogen carrier passes electrons to a carrier
electron acceptor. that accepts electrons, but not protons, the protons are released to
the outside of the cell (or intermembrane space of the mitochon-
2 H+
1/
2 O2 drion). The net effect of these processes is that the components of
the electron transport chain pump protons from one side of the
H2O membrane to the other, establishing the concentration gradient
across the membrane. Note that the gradient could not be estab-
FIGURE 6.18 Electron Transport As electrons are passed along lished if energy were not released during electron transfer.
the electron transport chain, the energy released is used to establish a
proton gradient. The Electron Transport Chain of Mitochondria
? O2 is serving as the terminal electron acceptor in this diagram; is it The electron transport chain of mitochondria has four different
being oxidized, or reduced? protein complexes, three of which function as proton pumps. In
addition, two electron carriers (ubiquinone and cytochrome c)
shuttle electrons between the complexes (figure 6.19):
in the case of mitochondria, from the matrix to the region between
■ Complex I (also called NADH dehydrogenase complex).
the inner and outer membranes. This ejection of protons creates a
This accepts electrons from NADH, ultimately transferring
proton gradient—an electrochemical gradient—across the mem-
them to ubiquinone (also called coenzyme Q); in the process,
brane. Energy of this gradient, proton motive force, is then har-
four protons are moved across the membrane.
vested by the cell to synthesize ATP. Recall from chapter 3 that
prokaryotes can also use the energy of proton motive force to ■ Complex II (also called succinate dehydrogenase complex).
transport substances and power the rotation of flagella. This accepts electrons from the TCA cycle, when FADH2 is
formed during the oxidation of succinate (see figure  6.17,
Components of an Electron Transport Chain step 6). Note that the electrons carried by FADH2 enter the
Most carriers in the electron transport chain are grouped into sev- electron transport chain “downstream” of those carried by
eral large protein complexes that function as proton pumps. Others NADH. Because of this, a pair of electrons carried by NADH
shuttle electrons from one complex to the next. Three general result in more protons being expelled than does a pair carried
groups of electron carriers are notable: quinones, cytochromes, by FADH2. Electrons are then transferred from complex II to
and flavoproteins. ubiquinone.
Quinones are lipid-soluble organic molecules that move ■ Complex III (also called cytochrome bc1 complex). This
freely in the membrane and can therefore transfer electrons accepts electrons from ubiquinone, which has carried them
between different protein complexes in the membrane. Several from either complex I or II. Complex III pumps four protons
types of quinones exist, one of the most common being ubiqui- across the membrane before transferring the electrons to
none (meaning “ubiquitous quinone”). Menaquinone, a quinone cytochrome c.
used in the electron transport chain of some prokaryotes, serves as ■ Complex IV (also called cytochrome c oxidase complex).
a source of vitamin K for humans and other mammals. This vita- This accepts electrons from cytochrome c and pumps two
min is required for proper blood coagulation, and mammals obtain protons across the membrane. Complex IV is a terminal oxi-
much of their requirement by absorbing menaquinone produced doreductase, meaning it transfers the electrons to the terminal
by bacteria growing in the intestinal tract. electron acceptor, which, in this case, is O2.
Cytochromes are proteins that contain heme, a molecule that
holds an iron atom in its center. Several different cytochromes exist, The Electron Transport Chains of Prokaryotes
each distinguished with a letter, for example, cytochrome c. The Considering the versatility and diversity of prokaryotes, it should
presence of certain cytochromes can be used as an identifying not be surprising that the types and arrangement of their electron
marker. For instance, the oxidase test—which is used in the steps to transport components vary tremendously. In fact, a single species
identify Neisseria, Pseudomonas, and Campylobacter species— can have several alternative carriers, allowing cells to cope with
detects the activity of cytochrome c oxidase (see table 10.5). ever-changing growth conditions.
144 Chapter 6 Metabolism: Fueling Cell Growth

2
Pentose phosphate
GLUCOSE

Glycolysis Yields
FIGURE 6.19 The Electron Transport Chain of Mitochondria The electrons carried by NADH
Reducing
pathway 1 P~ P ~ P +
Starts the oxidation of glucose
Oxidizes glucose to pyruvate
ATP
by substrate-level
phosphorylation
power

are passed to complex I. They are then passed to ubiquinone (coenzyme Q), which transfers them to
complex III. Cytochrome c then transfers electrons to complex IV. From there, they are passed to O2.
Electrons carried by FADH2 enter the chain at complex II, which then passes
Yields Reducing
power
them to ubiquinone; from there, the electrons follow the same path as the
Biosynthesis Fermentation
Eukaryotic cell ones donated by NADH. Protons are shuttled from the mitochondrial
5
Reduces pyruvate

Pyruvate Pyruvate
or a derivative

Acids, alcohols, and gases

matrix to the intermembrane space by complexes I, III, and IV, creating
3a Transition step
CO2 CO2
the proton motive force. ATP synthase allows protons to reenter the
Yields

mitochondrial matrix, harvesting the energy released to drive ATP

Reducing
power

Acetyl- Acetyl-
CoA CoA

synthesis.
x2 CO2

CO2
? Which would be expected to generate more ATP per
3b TCA cycle
Incorporates an acetyl
group and releases CO2
(TCA cycles twice) Inner electron carried—NADH or FADH2?
Yields
P~ P ~ P + Reducing
power
mitochondrial
ATP
by substrate-level
phosphorylation
4 Respiration
Uses the electron transport
chain to convert reducing
membrane
power to proton motive force

Yields
P~ P ~ P
ATP
by oxidative
phosphorylation

Electron Transport Chain Use of Proton Motive Force

Proton motive force

ATP synthase
Complex I Complex III Complex IV is used to drive: (ATP synthesis)

4 H+ 4 H +
2 H + 10 H+ Intermembrane
space
Ubiquinone
Ubiquin Cytochrome c

Path of 2 e–
electrons

Mitochondrial
2 H+ 1/
2
O2 matrix
Complex II
NADH Terminal
+ H2O electron acceptor
NAD+
H+

~ ~
3 ATP

~ + 3 Pi
3 ADP

The electron transport chain of E. coli provides an excellent four protons. The other ejects only two protons, but it can more ef-
example of the versatility of some prokaryotes. This bacterium fectively scavenge O2 and therefore is particularly useful in low O2
uses aerobic respiration when O2 is available, but in the absence of conditions.
O2 it can switch to anaerobic respiration if a suitable electron
acceptor such as nitrate is present. The E. coli electron transport Anaerobic Respiration Anaerobic respiration harvests less en-
chain serves as a model for both aerobic and anaerobic respiration. ergy than aerobic respiration, which makes sense considering the
lower electron affinities of other electron acceptors (see figure
Aerobic Respiration When growing aerobically in a glucose- 6.7). Some of the electron transport chain components used during
containing medium, E. coli can use two different NADH dehydro- anaerobic respiration are different from those of aerobic respira-
genases; one is a proton pump functionally equivalent to complex I tion. For example, E. coli can synthesize a terminal oxidoreduc-
of the mitochondrion (figure 6.20). E. coli also has a succinate de- tase that uses nitrate as a terminal electron acceptor when O2 is not
hydrogenase functionally equivalent to complex II of a mitochon- available. This produces nitrite, which E. coli then converts to
drion. In addition to these protein complexes, E. coli can produce ammonia, avoiding the toxic effects of nitrite. Other bacteria can
several alternatives, allowing the organism to optimally use a vari- reduce nitrate even further, forming compounds such as nitrous
ety of different energy sources, including H2. The bacterium does oxide (N2O), and nitrogen gas (N2).
not have the equivalent of complex III or cytochrome c. Instead, A group of obligate anaerobes called the sulfate-reducers use
quinones shuttle the electrons directly to one of two variations of a sulfate (SO42–) as a terminal electron acceptor, producing hydro-
ubiquinol oxidase, which are functionally equivalent to complex IV gen sulfide as an end product. The diversity and ecology of
of a mitochondrion, a terminal oxidoreductase. One variation of the sulfate-reducing bacteria will be discussed in chapter 11.
ubiquinol oxidase works optimally in high O2 conditions and expels sulfate-reducing bacteria, p. 258
 Part I Life and Death of Microorganisms 145

Prokaryotic cell

Cytoplasmic
membrane

Electron Transport Chain Uses of Proton Motive Force

ATP synthase Active transport Rotation of flagella

NADH dehydrogenase Ubiquinol oxidase (ATP synthesis) (one mechanism)
H+ (0 or 4) H+ (2 or 4) 10 H+ H+ H+
Proton motive force
Transported Outside of
is used to drive:
molecule cytoplasmic
Ubiquinone membrane

Path of 2 e–
electrons

Cytoplasm
1/ O2
2 H+ 2
Succinate
NADH dehydrogenase Terminal
+ electron acceptor
NAD+ H2O
H+

~ ~
3 ATP

~ + 3 Pi
3 ADP
FIGURE 6.20 The Electron Transport Chain of E. coli Growing Aerobically in a Glucose-Containing Medium The electrons carried
by NADH are passed to one of two different NADH dehydrogenases. They are then passed to ubiquinone, which transfers them to one of two
ubiquinol oxidases. From there they are passed to O2. The electrons carried by FADH2 enter the chain at succinate dehydrogenase, which then
transfers them to ubiquinone; from there, the electrons follow the same path as the ones donated by NADH. Protons are ejected by one of the
two NADH dehydrogenases and both ubiquinol oxidases, creating the proton motive force. ATP synthase allows protons to reenter the cell, using
the energy released to drive ATP synthesis. The proton motive force is also used to drive one form of active transport and to power the rotation
of flagella. E. coli has other components of the electron transport chain that function under different growth conditions.
? Succinate dehydrogenase is equivalent to which component of the mitochondrial electron transport chain?

ATP Synthase—Harvesting the Proton compared. It is not a straightforward comparison, however,

Motive Force to Synthesize ATP because oxidative phosphorylation has so many variables. This is
particularly true for prokaryotic cells because they use proton
Just as energy is required to establish a concentration gradient, energy motive force to drive processes other than ATP synthesis.
is released when the gradient is removed or reduced. The enzyme ATP Prokaryotic cells, as a group, use different carriers in their electron
synthase uses the energy of proton motive force to synthesize ATP. It transport chain and eject a variable number of protons per pair of
does this by allowing protons to flow back into the bacterial cell (or electrons passed.
matrix of the mitochondrion) in a controlled manner, simultaneously The basis for calculating the ATP yield of oxidative phos-
using the energy released to add a phosphate group to ADP. One mol- phorylation relies on experimental studies using rat mitochondria.
ecule of ATP is formed from the entry of approximately three protons. These studies indicate that approximately 2.5 ATP are made for
each pair of electrons transferred to the electron transport chain by
Theoretical ATP Yield of Oxidative NADH; about 1.5 ATP are made for each pair transferred by
Phosphorylation FADH2. For simplicity we will use whole numbers (3 ATP/NADH
By calculating the ATP yield of oxidative phosphorylation, and 2 ATP/FADH2) in calculations. Using these numbers, the
the relative energy gains of respiration and fermentation can be maximum theoretical energy yield for oxidative phosphorylation
146 Chapter 6 Metabolism: Fueling Cell Growth

in a prokaryotic cell (assuming the electron transport chain is simi- fate of the reducing power (NADH) generated during glycolysis.
lar to that of mitochondria) is: Recall that in eukaryotic cells, glycolysis takes place in the cyto-
From glycolysis: plasm, whereas the electron transport chain is located in the mito-
chondria. Consequently, the electrons carried by cytoplasmic NADH
■ 2 NADH→6 ATP (assuming 3 for each NADH) must be moved across the mitochondrial membrane before they can
From the transition step: enter the electron transport chain. This requires an expenditure of
approximately 1 ATP per NADH generated during glycolysis.
■ 2 NADH→6 ATP (assuming 3 for each NADH)

From the TCA cycle:

■ 6 NADH→18 ATP (assuming 3 for each NADH)
ATP Yield of Aerobic Respiration
in Prokaryotes
■ 2 FADH2→4 ATP (assuming 2 for each FADH2)
Now that the ATP-yielding components of the central metabolic
Total maximum ATP yield from oxidative phosphorylation = 34 pathways have been considered, we can calculate the theoretical
The ATP gain as a result of oxidative phosphorylation will be maximum ATP yield of aerobic respiration in prokaryotes. This
slightly less in eukaryotic cells than in prokaryotic cells because of the yield is illustrated in figure 6.21.
GLUCOSE

2
Pentose phosphate
pathway
Starts the oxidation of glucose
1
Glycolysis
Oxidizes glucose to pyruvate
Yields
~ ~ + Reducing
power
GLUCOSE
ATP
by substrate-level
phosphorylation

Glycolysis
Yields Reducing
power
Oxidizes glucose to pyruvate
Biosynthesis Fermentation
5
Reduces pyruvate
or a derivative

Pyruvate Pyruvate Acids, alcohols, and gases ~ ~

3a Transition step

Yields
Reducing
power
CO2 CO2
2 ATP
Acetyl-

net gain = 0
Acetyl-
CoA CoA

x2 CO2

CO2
~ ~
3b TCA cycle
Incorporates an acetyl
group and releases CO2
(TCA cycles twice)
2 ATP
Yields
~ ~ + Reducing
power
ATP
by substrate-level 4 Respiration
phosphorylation Uses the electron transport
chain to convert reducing
power to proton motive force

Yields
~ ~
ATP
by oxidative
phosphorylation

2 NADH ~ ~
Oxidative
phosphorylation 6 ATP

~ ~
Substrate-level
phosphorylation 2 ATP

Pyruvate Pyruvate

2 NADH ~ ~
Oxidative
phosphorylation 6 ATP
Acetyl- Acetyl-
CoA CoA

FIGURE 6.21 Maximum Theoretical

Energy Yield from Aerobic 6 NADH ~ ~
Respiration in a Prokaryotic Cell This Oxidative
x2 CO2 phosphorylation 18 ATP
maximum energy yield calculation
assumes that for every pair of electrons 2 FADH2 ~ ~
transferred to the electron transport Oxidative
chain, 3 ATP are synthesized; and for phosphorylation 4 ATP
CO2
every pair of electrons donated by
FADH2, 2 ATP are synthesized. TCA cycle ~ ~
Incorporates an acetyl
? Why is it difficult to calculate
group and releases CO2
Substrate-level
2 ATP
the actual maximum ATP yield of phosphorylation
(TCA cycles twice)
respiration in a prokaryotic cell?
 Part I Life and Death of Microorganisms 147

Substrate-level phosphorylation: beverages are produced using fermentations. Important end prod-
ucts of fermentation pathways include:
■ 2 ATP (from glycolysis; net gain)
■ 2 ATP (from the TCA cycle) ■ Lactic acid. Lactic acid (the ionized form is lactate) is pro-
duced when pyruvate itself serves as the terminal electron
■ 4 total
acceptor. Lactic acid and other end products of a group
Oxidative phosphorylation: of Gram-positive organisms called lactic acid bacteria are
instrumental in creating the flavor and texture of cheese,
■ 6 ATP (from the reducing power gained in glycolysis)
yogurt, pickles, cured sausages, and other foods. Yet lactic
■ 6 ATP (from the reducing power gained in the transition step) acid also causes food spoilage, and contributes to tooth decay
■ 22 ATP (from the reducing power gained in the TCA cycle) when produced by bacteria living on the teeth. Some animal
■ 34 total cells use this fermentation pathway temporarily when O2 is
in short supply; the accumulation of lactic acid in muscle
Total ATP gain (theoretical maximum) = 38 tissue causes the pain and fatigue associated with strenuous
exercise. lactic acid bacteria, p. 258 cheese, yogurt, and other fer-
MicroAssessment 6.4 mented milk products, p. 751 pickled vegetables, p. 752 fermented
Respiration uses the NADH and FADH2 generated in glycolysis, the meat products, p. 752
transition step, and the TCA cycle to synthesize ATP. The electron
transport chain is used to convert reducing power into proton motive GLUCOSE

Pentose phosphate Yields

Glycolysis

force. ATP synthase then harvests that energy to synthesize ATP.

2 Reducing
pathway 1
Oxidizes glucose to pyruvate P~ P ~ P + power
Starts the oxidation of glucose
ATP
by substrate-level
phosphorylation

The overall process is called oxidative phosphorylation. In aerobic

respiration, O2 serves as the terminal electron acceptor; anaerobic
respiration uses a molecule other than O2. Yields Reducing
power

10. In bacteria, what is the role of the molecule that serves as a Biosynthesis
5 Fermentation
Reduces pyruvate
or a derivative

source of vitamin K for humans? 3a Transition step

Pyruvate Pyruvate Acids, alcohols, and gases

CO2 CO2

11. Why is the overall ATP yield in aerobic respiration only a

Yields
Reducing
power

Acetyl- Acetyl-
CoA CoA

theoretical number?
12. Why could an oxidase also be called a reductase? +
x2 CO2

CO2

3b TCA cycle
Incorporates an acetyl
group and releases CO2
(TCA cycles twice)

Yields
P~ P ~ P + Reducing
power
ATP
by substrate-level 4 Respiration

6.5 Fermentation
Uses the electron transport

■
phosphorylation
chain to convert reducing
power to proton motive force

Yields
P~ P ~ P
ATP
by oxidative
phosphorylation

Learning Outcome
16. Describe the role of fermentation and the importance of the NADH + H+ NAD+
common end products. O O OH O
–
H3C C C O H3C C C O–
Fermentation is used by organisms that cannot respire, either
because a suitable inorganic terminal electron acceptor is not
H
available or because they lack an electron transport chain. E. coli
Pyruvate Lactate
is a facultative anaerobe able to use any of three ATP-generating
options: aerobic respiration, anaerobic respiration, and fermenta- (a) Lactic acid fermentation
tion. In contrast, the only option for Streptococcus pneumoniae
is fermentation because it does not have an electron transport
CO2
chain. NADH + H+ NAD+
In general, the only ATP-generating reactions of fermenta- O O O OH
tion are those of glycolysis and involve substrate-level phos-
H3C C C O– H3C C H H3C C H
phorylation. The additional steps simply consume excess reducing
power as a way of regenerating NAD+ (figure  6.22). This is a H
critical function, because NAD+ is needed to accept electrons in Pyruvate Acetaldehyde Ethanol
subsequent rounds of glycolysis—without it, glycolysis would
(b) Ethanol fermentation
stop. To consume reducing power, fermentation uses an organic
compound such as pyruvate or a derivative as a terminal electron FIGURE 6.22 Fermentation Pathways Use Pyruvate or a
Derivative as a Terminal Electron Acceptor (a) In lactic acid
acceptor.
fermentation, the pyruvate generated during glycolysis serves as
The end products of fermentation are significant for a number the terminal electron acceptor, producing lactate. (b) In ethanol
of reasons (figure 6.23). For one thing, certain end products help fermentation, the pyruvate is first converted to acetaldehyde, which
identify bacterial isolates because a given organism uses a charac- then serves as the terminal electron acceptor, producing ethanol.
teristic fermentation pathway. In addition, some end products are ? Why is it important for cells to have a mechanism to oxidize
commercially valuable. Chapter 31 describes how foods and NADH?
148 Chapter 6 Metabolism: Fueling Cell Growth

■ Ethanol. Ethanol is produced in a pathway that first removes holes, and propionic acid gives the cheese its unique flavor.
CO2 from pyruvate, generating acetaldehyde, which then cheese, p. 751
serves as the terminal electron acceptor. The end products of
these sequential reactions—which are used in making wine, ■ Mixed acids. These are produced in a multistep branching
beer, spirits, and bread—are ethanol and CO2, (see figures pathway, generating a variety of different fermentation prod-
31.4, 31.5, and 31.6). Ethanol is also an important biofuel. ucts including lactic acid, succinic acid (the ionized form is
Saccharomyces (yeast) and Zymomonas (bacteria) use this succinate), ethanol, acetic acid (the ionized form is acetate), and
pathway. wine, p.  753 beer, p.  754 distilled spirits, p.  755 gases. This is another pathway used to differentiate members of
bread, p. 755 the family Enterobacteriaceae; the methyl-red test detects the
low pH resulting from the acidic end products, distinguishing
■ Butyric acid. Butyric acid (the ionized form is butyrate) and
members that use this pathway, such as E. coli, from those that
a variety of other end products are produced in a complex
do not, such as Klebsiella and Enterobacter (see table  10.5).
multistep pathway used by Clostridium species, which are
methyl-red test, p. 243
obligate anaerobes. Under certain conditions, some organ-
isms use a variation of this pathway to produce the organic ■ 2,3-Butanediol. 2,3-Butanediol is produced in a multistep
solvents butanol and acetone. pathway that uses two molecules of pyruvate to generate
■ Propionic acid. Propionic acid (the ionized form is propio- acetoin and two molecules of CO2. Acetoin is then used as
nate) is generated in a multistep pathway that first adds CO2 the terminal electron acceptor. The primary significance of
to pyruvate, generating a compound that then serves as a ter- this pathway is that it serves to differentiate certain members
minal electron acceptor. After NADH reduces this, it is fur- of the family Enterobacteriaceae. The Voges-Proskauer test
ther modified to form propionate. Propionibacterium species detects acetoin, distinguishing members that use this pathway
use this pathway, and their growth is encouraged as a part (such as Klebsiella and Enterobacter) from those that do not
of Swiss cheese production. The CO2 they make forms the (such as E. coli; see table 10.5). Voges-Proskauer test, p. 243
GLUCOSE

FIGURE 6.23 End Products

MicroAssessment 6.5
Pentose phosphate Yields
2 Glycolysis Reducing
pathway 1
Oxidizes glucose to pyruvate P~ P ~ P + power
Starts the oxidation of glucose
ATP
by substrate-level
phosphorylation of Fermentation Pathways
Fermentation stops short of the TCA cycle, using pyruvate or a
? How can end products of
fermentation help identify
derivative as a terminal electron acceptor. Many end products of
Yields Reducing
power

a bacterium? fermentation are commercially valuable.

Biosynthesis Fermentation
5
Reduces pyruvate
or a derivative
13. Why would a cell ferment rather than respire?
Pyruvate Pyruvate Acids, alcohols, and gases

3a Transition step

Yields
Reducing
CO2 CO2
14. How do the methyl-red and Voges-Proskauer tests differentiate
power

Acetyl-
CoA
Acetyl-
CoA between certain members of the Enterobacteriaceae?
x2 CO2
15. Fermentation is used as a means of preserving foods. Why would
CO2 it slow spoilage? +
3b TCA cycle
Incorporates an acetyl
group and releases CO2
(TCA cycles twice)

Yields
P~ P ~ P + Reducing
power
ATP
by substrate-level 4 Respiration
phosphorylation Uses the electron transport
chain to convert reducing
power to proton motive force

Yields
P~ P ~ P
ATP
by oxidative
phosphorylation

Pyruvate

Fermentation
pathway Lactic acid Ethanol Butyric acid Propionic acid Mixed acids 2,3-Butanediol

Streptococcus
Microorganisms Lactobacillus Saccharomyces Clostridium Propionibacterium E. coli Enterobacter

End products Lactic acid Ethanol Butyric acid Propionic acid Acetic acid Formic acid
CO2 Butanol Acetic acid Lactic acid Ethanol
Acetone CO2 Succinic acid Lactic acid
Isopropanol Ethanol 2,3-Butanediol
CO2 CO2 CO2
H2 H2 H2
 Part I Life and Death of Microorganisms 149

6.6 ■ Catabolism of Organic Microbes can use a variety of organic compounds other than glu-
cose as energy sources, including polysaccharides, proteins and
Compounds Other lipids. To break these down into their respective sugar, amino
Than Glucose acid, and lipid subunits, cells synthesize hydrolytic enzymes,
which break bonds by adding water. To use a macromolecule in
Learning Outcome the surrounding medium, a cell secretes the appropriate hydrolytic
enzyme and then transports the resulting subunits into the cell (see
17. Briefly describe how polysaccharides and disaccharides, lipids,
and proteins are degraded and utilized by a cell. figure 3.30). Inside the cell, the subunits are further degraded to
form appropriate precursor metabolites (figure 6.24). Recall that

POLYSACCHARIDES DISACCHARIDES LIPIDS (fats) PROTEINS

Starch Lactose Maltose
Cellulose Sucrose lipases proteases

amylases cellulases disaccharidases glycerol amino acids

+
deamination
monosaccharides GLUCOSE fatty acids
(simple sugars)
NH3
Pentose phosphate
pathway Glycolysis

Applies to
both branches
in glycolysis

ß-oxidation
removes
Pyruvate Pyruvate 2-carbon units.

Acetyl- Acetyl-
CoA CoA

FIGURE 6.24 Catabolism of Organic Compounds x2

Other Than Glucose The subunits of macromolecules
are degraded to form the appropriate precursor
metabolites. These metabolites can then either be
oxidized in one of the central metabolic pathways or be
used in anabolism.
? Which is more common—an organism that produces TCA cycle
amylase or one that produces cellulase?
150 Chapter 6 Metabolism: Fueling Cell Growth

precursor metabolites can be either oxidized in one of the central

MicroAssessment 6.6
metabolic pathways or used in biosynthesis.  hydrolysis, p. 24
In order for polysaccharides, lipids, and proteins to be used as energy
sources, they must be first hydrolyzed to release their respective
Polysaccharides and Disaccharides subunits. These are then converted to the appropriate precursor
metabolites so they can enter a central metabolic pathway.
Starch and cellulose are both polymers of glucose, but different
types of chemical linkages join their subunits. The nature of this 16. Why do cells secrete hydrolytic enzymes?
difference profoundly affects their degradation. Enzymes called 17. Explain the process used to degrade fatty acids.
amylases are made by a wide variety of organisms to digest 18. How would cellulose-degrading bacteria in the rumen of a cow
starches. In contrast, cellulose is broken down by cellulases, which benefit the animal? +
are produced by relatively few organisms. Bacteria that reside in
the rumen of animals, as well as many types of fungi, are among
those that can produce cellulase. Considering that cellulose is the 6.7 ■ Chemolithotrophs
most abundant organic compound on earth, it is not surprising that
fungi are important decomposers in terrestrial habitats. The glucose Learning Outcome
subunits released when polysaccharides are hydrolyzed can then 18. Explain how chemolithotrophs obtain energy.
enter glycolysis to be oxidized to pyruvate.  starch, p. 32 cellulose,
p. 32 rumen, p. 732 Prokaryotes as a group are unique in their ability to use reduced
Disaccharides including lactose, maltose, and sucrose are inorganic chemicals such as hydrogen sulfide (H2S) and ammonia
hydrolyzed by specific disaccharidases. For example, the enzyme (NH3) as sources of energy. Note that these compounds are the
β-galactosidase breaks down lactose, forming glucose and galac- very ones produced as a result of anaerobic respiration, when
tose. Glucose can enter glycolysis directly, but other monosac- inorganic molecules, such as sulfate and nitrate, serve as terminal
charides must first be converted to one of the precursor metabolites. electron acceptors. This is one important example of how nutrients
disaccharides, p. 31 are cycled; the waste products of one organism serve as an energy
source for another. biogeochemical cycling and energy flow, p. 725
Lipids Chemolithotrophs fall into four general groups (table 6.7):
Fats, the most common simple lipids, are a combination of fatty ■ Hydrogen bacteria oxidize hydrogen gas.
acids and glycerol. Fats are hydrolyzed by lipases. The glycerol
■ Sulfur bacteria oxidize hydrogen sulfide.
component is then converted to the precursor metabolite dihy-
droxyacetone phosphate, which enters glycolysis. The fatty acids ■ Iron bacteria oxidize reduced forms of iron.
are degraded using a series of reactions collectively called ■ Nitrifying bacteria include two groups of bacteria: one oxi-
β-oxidation. Each sequential reaction transfers a 2-carbon unit dizes ammonia (forming nitrite), and the other oxidizes nitrite
from the end of the fatty acid to coenzyme A, forming acetyl-CoA, (producing nitrate).
which enters the TCA cycle. Each β-oxidation is a redox reaction,
Chemolithotrophs extract electrons from inorganic energy sources,
generating 1 NADH + H+ and 1 FADH2. simple lipids, p. 33
passing them to an electron transport chain that generates a proton
motive force. The energy of this gradient is then harvested
Proteins to make ATP, using the processes described earlier. As with chemo-
Proteins are hydrolyzed by proteases, which break peptide bonds heterotrophs, the amount of energy gained in metabolism depends on
between amino acid subunits. The amino group of the resulting the energy source and the terminal electron acceptor (see figure 6.7).
amino acids is removed by a reaction called a deamination. The Chemolithotrophs generally thrive in very specific environ-
remaining carbon skeletons are then converted into the appropri- ments where reduced inorganic compounds are found. For exam-
ate precursor molecules. protein, p. 25 ple, certain bacteria are found in sulfur-rich acidic environments;

TABLE 6.7 Metabolism of Chemolithotrophs

Common Name Source Oxidation Reaction(s) Important Feature(s) Common Genera
of Organism of Energy (Energy Yielding) of Group in Group

Hydrogen bacteria H2 H2 + * O2→H2O Can also use simple organic Hydrogenomonas

compounds for energy
Sulfur bacteria H2S H2S + * O2→H2O + S Some members of this group can Acidithiobacillus,
(non-photosynthetic) live at a pH of less than 1. Thiobacillus, Beggiatoa,
S + 1* O2 + H2O→H2SO4
Thiothrix
Iron bacteria Reduced 2 Fe2+ + * O2 + H2O→2 Fe3+ + 2 OH– Iron oxide present in the sheaths Sphaerotilus, Gallionella
Iron (Fe2+) of these bacteria
Nitrifying bacteria NH3 NH3 + 1* O2→HNO2 + H2O Important in the nitrogen cycle Nitrosomonas
Important in the nitrogen cycle
HNO2 HNO2 + * O2→HNO3 Nitrobacter
 Part I Life and Death of Microorganisms 151

PERSPECTIVE 6.1
Mining with Microbes
Microorganisms have been used for thou- activities are expensive and harmful to the all copper produced in the world comes from
sands of years in the production of bread environment. With the development of the process of biomining. Similar processes
and wine. Only in the past several decades, biomining, some of these problems are being are being applied to gold mining.
however, have microorganisms been used solved. The current process of biomining uses
with increasing frequency in another area— In the process of biomining copper, the microbes naturally in the ore. Many improve-
the mining industry. The mining process low-grade ore is piled outside the mine and ments should be possible. For example, oxidat-
traditionally consists of digging crude ores then treated with acid. The acidic conditions ing the minerals generates heat to the point that
(mineral-containing rocks) from the earth, encourage the growth of Acidithiobacillus spe- the bacteria may be killed, but it may be pos-
crushing them, and then extracting the desired cies present naturally in the ore. These acido- sible to use thermophiles to overcome this
minerals from the contaminants. The extrac- philic bacteria use CO2 as a source of carbon problem. In addition, many ores contain heavy
tion process for copper and gold frequently and gain energy by oxidizing sulfides of iron metals, such as mercury, cadmium, and arse-
involves harsh conditions, such as smelting, first to sulfur and then to sulfuric acid. The nic, which are toxic to the bacteria, but perhaps
and burning off the contaminants before sulfuric acid dissolves the insoluble copper microorganisms resistant to these metals could
extracting the metal with cyanide. Such and gold from the ore. Currently about 25% of be found. Biomining is still in its infancy.

these organisms can be used to enhance the recovery of metals capture radiant energy and convert it to chemical energy in the form
because they oxidize metal sulfides (see Perspective 6.1). of ATP. An unrelated set of reactions (sometimes called the light-
Thermophilic chemolithotrophs that grow near hydrothermal independent reactions or dark reactions) uses the ATP to synthesize
vents of the deep ocean obtain energy from reduced inorganic organic compounds. This involves carbon fixation, which converts
compounds that spew from the vents. The diversity and ecology of CO2 into organic compounds. We will describe the steps of carbon
some chemolithotrophs will be discussed in chapter 11. fixation in a separate section later in the chapter because chemolitho-
Unlike organisms that use organic molecules to fill both their trophs use the process as well. Characteristics of various photosyn-
energy and carbon needs, chemolithotrophs incorporate CO2 into an thetic mechanisms are summarized in table 6.8.
organic form. This process (carbon fixation) will be described later.
Capturing Radiant Energy
MicroAssessment 6.7 Photosynthetic organisms are highly visible in their natural habitats
Chemolithotrophs use reduced inorganic compounds as an energy because they have various colored pigments that capture the energy of
source. They use carbon dioxide as a carbon source. light (radiant energy). The colors we observe are due to the wave-
19. Describe the roles of hydrogen sulfide and carbon dioxide in lengths reflected by the pigments—for example, pigments that absorb
chemolithoautotrophic metabolism. only blue and red light are green (see figure 5.6). Multiple pigments
20. Which energy source, Fe2+ or H2S, would result in the greatest are involved in photosynthesis, increasing the range of wavelengths
energy yield when O2 is used as a terminal electron acceptor absorbed by a cell. The pigments are located in protein complexes
(hint: refer to figure 6.7)? + called photosystems within photosynthetic membranes (figure 6.25).
The photosystems specialize in capturing and using the energy of light.

6.8 ■ Photosynthesis Electron

Photosystem
transport chain
Learning Outcomes Radiant Reaction-center
19. Describe the role of chlorophylls, bacteriochlorophylls, accessory energy chlorophyll e–
pigments, reaction-center pigments, and antennae pigments in
Chlorophyll
capturing radiant energy. molecule
20. Compare and contrast the tandem photosystems of cyanobacteria
and photosynthetic eukaryotes with the single photosystems of
purple and green bacteria.

Plants, algae, and several groups of bacteria harvest the radiant

energy of sunlight, and then use it to power the synthesis of
organic compounds from CO2. The capture and subsequent con-
version of radiant energy into chemical energy is called
photosynthesis. The general reaction—with X indicating an ele- Photosynthetic membrane
ment such as oxygen or sulfur—is as follows:
FIGURE 6.25 Photosystem Chlorophyll and other pigments
Light Energy
capture the energy of light and then transfer it to reaction-center
6 CO2 + 12 H2X → C6H12O6 + 12 X + 6 H2O chlorophyll, which emits an electron that is then passed to an electron
Photosynthetic processes are generally considered in two distinct transport chain.
stages. The light reactions (also called the light-dependent reactions) ? What is the role of an electron transport chain?
152 Chapter 6 Metabolism: Fueling Cell Growth

TABLE 6.8 Comparison of the Photosynthetic Mechanisms Used by Different Organisms

Oxygenic Photosynthesis Anoxygenic Photosynthesis

Plants, Algae Cyanobacteria Purple Bacteria Green Bacteria

Location of the In membranes of In membranes of Within the cytoplasmic Primarily within the
photosystem thylakoids, which are thylakoids, located within membrane; extensive cytoplasmic membrane;
within the stroma of the cell invaginations in that chlorosomes attached to
chloroplasts membrane increase the the inner surface of the
surface area. membrane contain the
accessory pigments.
Type of photosystem Photosystem I and photosystem II Similar to photosystem II Similar to photosystem I
Primary light-harvesting Chlorophyll a Chlorophyll a Bacteriochlorophylls Bacteriochlorophylls
pigment
Mechanism for generating Non-cyclic photophosphorylation using both Reversed electron transport Non-cyclic use of the
reducing power photosystems photosystem
Source of electrons for H2O H2O Varies among the organisms in the group; may include
reducing power H2S, H2, or organic compounds.
CO2 fixation Calvin cycle Calvin cycle Calvin cycle Reversed TCA cycle
Accessory pigments Carotenoids Carotenoids, phycobilins Carotenoids Carotenoids

Photosynthetic pigments include chlorophylls, bacteriochlo- The photosystems of the purple and green bacteria are embed-
rophylls, and accessory pigments. Chlorophylls are found in ded in the cytoplasmic membrane. Purple bacteria have extensive
plants, algae, and cyanobacteria. The various types of chloro- invaginations in the membrane that maximize the surface area.
phylls are designated with a letter following the term—for Green bacteria have specialized structures called chlorosomes
example, chlorophyll a. Bacteriochlorophylls are found in attached to the inner surface of the cytoplasmic membrane. These
anoxygenic photosynthetic bacteria (“anoxygenic” means they do structures contain the accessory pigments.
not generate O2). These pigments absorb wavelengths not
absorbed by chlorophylls, allowing the bacteria to grow in habi-
tats where other photosynthetic organisms cannot. Accessory Converting Radiant Energy
pigments increase the efficiency of light capture by absorbing
wavelengths not absorbed by the other pigments. These pigments
into Chemical Energy
include carotenoids—found in a wide variety of photosynthetic Photosynthetic organisms use the light-dependent reactions to
prokaryotes and eukaryotes—and phycobilins, which are unique accomplish two tasks. First, they use radiant energy to fuel ATP
to cyanobacteria and red algae. synthesis, the process of photophosphorylation. They also need
Within the photosystems, some pigments function as to generate reducing power so they can fix CO2. Depending on the
reaction-center pigments and others function as antennae pig- method used to fix CO2, the type of reducing power required may
ments (figure  6.25). Reaction-center pigments are electron be either NADPH or NADH.
donors in the photosynthetic process. When excited by radiant
energy, these emit high-energy electrons, which are then passed to Light-Dependent Reactions in Cyanobacteria
an electron transport chain similar to that used in respiration. The and Photosynthetic Eukaryotic Cells
reaction-center pigment of oxygenic photosynthetic organisms Cyanobacteria and chloroplasts have two distinct photosystems
(plants, algae, and cyanobacteria) is chlorophyll a, whereas the that work in tandem (figure  6.26). The sequential absorption of
anoxygenic photosynthetic organisms (purple and green bacteria) energy by the two photosystems allows the process to raise the
use one of the bacteriochlorophylls. Antennae pigments make up energy of electrons stripped from water to a high enough level to
a complex that acts as a funnel, capturing the energy of light and be used to generate a proton motive force as well as produce
then transferring it to the reaction-center pigment. reducing power. The process is oxygenic—that is, it generates O2.
The photosystems of cyanobacteria are embedded in the First we will consider the simplest situation, which occurs
membranes of structures called thylakoids located within the when the cell needs to synthesize ATP, but not reducing power
cells. Plants and algae also have thylakoids, in the stroma of (NADPH). To accomplish this, only photosystem I is used. When
the chloroplast (see  figure 3.53). The similarity between the radiant energy is absorbed by this photosystem, the reaction-center
structure of chloroplasts and cyanobacteria is not surprising chlorophylls emit high-energy electrons. The electrons are then
considering that the organelle appears to have descended passed to an electron carrier, which transports them to a proton
from an ancestor of a cyanobacterium (see Perspective 3.1). pump similar to complex III in the electron transport chain of
thylakoid, p. 76 stroma, p. 76 mitochondria. After being used to move protons across the
 Part I Life and Death of Microorganisms 153

Excited
chlorophyll Electron
Proton gradient carrier
formed for ATP
synthesis e–
Excited NADP
chlorophyll H+ e– reductase
Electron
carrier
NADPH
e– NADP+
Reaction-
Energy of electrons

Proton
pump center
Electron chlorophyll
carrier
Reaction-
e– Radiant
center
chlorophyll energy

Water-splitting
enzyme
Radiant
energy Z 2 H2O
e–
4 H+ + O2

Photosystem II Proton pump Photosystem I NADP reductase

FIGURE 6.26 The Tandem Photosystems of Cyanobacteria and Chloroplasts Radiant energy captured by photosynthetic pigments
excites the reaction-center chlorophyll, causing it to emit a high-energy electron, which is then passed to an electron transport chain. In cyclic
photophosphorylation, electrons emitted by photosystem I are returned to that photosystem; the path of the electrons is shown in green arrows.
In non-cyclic photophosphorylation, the electrons used to replenish photosystem I are donated by radiant energy–excited photosystem II; the path
of these electrons is shown in orange arrows. In turn, photosystem II replenishes its own electrons by stripping them from water, producing O2.
? When do the cells need to use non-cyclic photophosphorylation?

membrane—generating a proton motive force—the electrons are from water molecules to a high enough level that they can be used
returned to photosystem I. As in oxidative phosphorylation, ATP syn- to power photophosphorylation. Photosystem I then accepts those
thase harvests the energy of the proton motive force to synthesize electrons, which still have some energy, and again captures the
ATP. This overall process is called cyclic photophosphorylation energy of light to boost the energy of the electrons to an even
because the electrons have followed a cyclical path—the molecule that higher level necessary to reduce NADPH.
serves as the electron donor (reaction-center chlorophyll) is also the
terminal electron acceptor. Light-Dependent Reactions in Anoxygenic
When photosynthetic cells must produce both ATP and reduc- Photosynthetic Bacteria
ing power, non-cyclic photophosphorylation is used. In this pro- Anoxygenic photosynthetic bacteria have only a single photosys-
cess, the electrons emitted by photosystem I are not passed to the tem and cannot use water as an electron donor for reducing
proton pump but instead reduce NADP+ to NADPH. Although this power. This is why they are anoxygenic (do not generate O2).
action provides reducing power, the cell must now use another These bacteria use electron donors such as hydrogen gas (H2),
source to replenish the electrons emitted by reaction-center chloro- hydrogen sulfide (H2S), and organic compounds. Two groups of
phyll. In addition, the cell still needs to generate a proton motive anoxygenic photosynthetic bacteria are the purple bacteria and
force in order to synthesize ATP. Photosystem II plays a critical green bacteria. purple bacteria, p. 259 green bacteria, p. 260
role in this process. When photosystem II absorbs radiant energy, Purple bacteria synthesize ATP using a photosystem similar
the reaction-center chlorophylls emit high-energy electrons that to photosystem II of cyanobacteria and eukaryotes. However, this
can be donated to photosystem I. First, however, the electrons are photosystem does not raise the electrons to a high enough energy
passed to the proton pump, which uses some of their energy to level to reduce NAD+ (or NADP+), so the cells must use an alter-
establish the proton motive force. The electrons emitted from pho- native mechanism to generate reducing power. They use a process
tosystem II are replenished when an enzyme within that complex called reversed electron transport, expending ATP to run the elec-
extracts electrons from water, donating them to the reaction-center tron transport chain in the reverse direction, or “uphill.”
chlorophyll. Removal of electrons from two molecules of water Green bacteria have a photosystem similar to photosystem I.
generates O2. In essence, photosystem II captures the energy of The electrons emitted from this can either generate a proton
light and then uses it to raise the energy level of electrons stripped motive force or reduce NAD+.
154 Chapter 6 Metabolism: Fueling Cell Growth

MicroAssessment 6.8 Calvin Cycle

Photosynthetic organisms harvest radiant energy and use it to power the The Calvin cycle, or Calvin-Benson cycle, named in honor of the
synthesis of organic compounds from CO2. Various pigments are used scientists who described much of it, is a complex cycle. It can be
to capture radiant energy. These pigments are arranged in complexes viewed as having three essential stages—incorporation of CO2 into
called photosystems. When reaction-center chlorophyll absorbs the an organic compound, reduction of the resulting molecule, and
energy of light, a high-energy electron is emitted. This is then passed
regeneration of the starting compound (figure 6.27). Because of the
along an electron transport chain to generate a proton motive force,
which is used to synthesize ATP. Plants and cyanobacteria use water as complexities, it is easiest to consider the process as consisting of six
a source of electrons for reducing power, generating O2. Anoxygenic “turns” of the cycle. Together, these six turns generate a net gain of
photosynthetic bacteria obtain electrons from a reduced compound two molecules of glyceraldehyde 3-phosphate (G3P), which can be
other than water and do not evolve O2. converted into one molecule of fructose 6-phosphate, an intermedi-
21. β-carotene is a carotenoid that mammals can use as a source of ate of glycolysis. The three stages of the Calvin cycle are:
vitamin A. What is the function of carotenoids in photosynthetic ■ Stage 1. 1 Carbon dioxide enters the cycle when an enzyme
organisms? commonly called rubisco (ribulose bisphosphate carboxylase)joins
22. What is the advantage of having tandem photosystems? it to a 5-carbon compound, ribulose 1,5-bisphosphate (RuBP). The
23. Energy is required to reverse the flow of the electron transport resulting compound spontaneously hydrolyzes to produce two
chain. Why would this be so? + molecules of a 3-carbon compound, 3-phosphoglycerate (3PG).
■ Stage 2. 2 A sequential input of energy (ATP) and reducing
power (NADPH) is used in steps that, together, convert 3PG
6.9 ■ Carbon Fixation to G3P. This molecule is also a precursor metabolite formed as
an intermediate in glycolysis and can have a variety of different
Learning Outcome fates. It can be used in biosynthesis, oxidized to make other pre-
21. Describe the three stages of the Calvin cycle. cursor compounds, or converted to a 6-carbon sugar. A critical
aspect of the Calvin cycle, however, is that RuBP must be regen-
Chemolithoautotrophs and photoautotrophs use carbon dioxide (CO2) erated from G3P for the cycle to continue. Consequently, in six
to synthesize organic compounds, the process of carbon fixation. In turns of the cycle, a maximum of 2 G3P can be converted to a
photosynthetic organisms, this process is called the light-independent 6-carbon sugar; the rest is used to regenerate RuBP.
reactions. Carbon fixation consumes a great deal of ATP and reducing ■ Stage 3. 3 Many of the steps used to regenerate RuBP involve
power, which should not be surprising considering that the reverse reactions of the pentose phosphate cycle.
process (oxidizing those same
compounds to CO2) liberates a
great deal of energy. The 6 CO2
1 Carbon dioxide is added to ribulose
Calvin cycle is the most com- 1,5-bisphosphate to start a new
mon pathway used to fix car- round of the cycle.
12 molecules
bon, but some prokaryotes 3-phosphoglycerate
incorporate CO2 using other
6 molecules ~ ~
mechanisms. For example, the ribulose 1,5-bisphosphate 12 ATP
green bacteria and some mem- STAGE 1
bers of the Archaea use a
~
pathway that reverses the steps
12 ADP
of the TCA cycle.
12 molecules
~ 1,3-bisphosphoglycerate
STAGE 3 ~
6 ADP
STAGE 2 12 NADPH + H+
~ ~
6 ATP
6 molecules
ribulose 5-phosphate 12 molecules
glyceraldehyde 12 NADP+
FIGURE 6.27 The Calvin 3-phosphate
Cycle The Calvin cycle has three Series 12 Pi
essential stages: (1) incorporation 3 Ribulose 1,5-bisphosphate is of complex
of CO2 into an organic compound; regenerated so that the cycle reactions
2 ATP and NADPH are used to reduce
(2) reduction of the resulting can continue. the product of stage 1, producing
1 molecule
molecule; and (3) regeneration of e
fructose 6-phosphate glyceraldehyde 3-phosphate, which
the starting compound. can be used in biosynthesis.

? How much ATP and NADPH

must be spent to synthesize
one molecule of fructose? Cell components
 Part I Life and Death of Microorganisms 155

Yield of the Calvin Cycle 6.10 ■ Anabolic Pathways—

One molecule of the 6-carbon sugar fructose can be generated for
every six turns of the cycle. These six turns consume 18 ATP and
Synthesizing Subunits
12 NADPH + H+. from Precursor Molecules
Learning Outcome
MicroByte
Although rubisco is unique to autotrophs, it is probably the most 22. Describe the synthesis of lipids, amino acids, and nucleotides.
abundant enzyme on earth!
Prokaryotes, as a group, are highly diverse with respect to the com-
pounds they use for energy but remarkably similar in their biosynthetic
MicroAssessment 6.9 processes. They synthesize the necessary subunits, using specific ana-
The process of carbon fixation consumes a great deal of ATP and bolic pathways that require ATP, reducing power in the form of
reducing power. The Calvin cycle is the most common pathway used NADPH, and the precursor metabolites formed in the central meta-
to incorporate inorganic carbon into an organic form. bolic pathways (figure 6.28). Organisms lacking one or more enzymes
24. What is the role of rubisco? in a given biosynthetic pathway must have the end product provided
from an external source. This is why fastidious bacteria, such as lactic
25. What would happen if ribulose 1,5-bisphosphate (RuBP)
were depleted in a cell? + acid bacteria, require many different growth factors. Once the subunits
are synthesized or transported into the cell, they can be assembled to

Pentose phosphate Glycolysis

pathway

Glucose 6-phosphate
Lipopolysaccharide
(polysaccharide)
Fructose 6-phosphate
Ribose 5-phosphate
Nucleotides Peptidoglycan
amino acids Dihydroxyacetone
Erythrose 5-phosphate (histidine) phosphate
Amino acids Lipids
(phenylalanine, (glycerol
tryptophan, component)
tyrosine)
3-phosphoglycerate
Amino acids
(cysteine,
glycine, serine)
Phosphoenolpyruvate
Amino acids
(phenylalanine,
tryptophan, tyrosine)
Pyruvate Pyruvate Amino acids
(alanine,
leucine, valine)

Acetyl-CoA Acetyl-CoA
Lipids
(fatty acids)

Oxaloacetate
FIGURE 6.28 The Use of Precursor
Metabolites in Biosynthesis The size of Amino acids
the arrows indicates the relative quantity (aspartate, asparagine, x2 α-ketoglutarate
of each precursor metabolite needed to isoleucine, lysine, Amino acids
produce a given weight of E. coli cells. methionine, threonine) (arginine, glutamate,
glutamine, proline)
? If an E. coli cell lost the ability to use
the pentose phosphate pathway, which
TCA cycle
amino acids would it require as growth
factors?
156 Chapter 6 Metabolism: Fueling Cell Growth

make macromolecules. Various different macromole- NH2

α-ketoglutarate NH3 (ammonia)
cules can then be joined to form the structures that
make up the cell. fastidious, p. 93 Glutamate is
synthesized
Lipid Synthesis Aspartate by adding ammonia
to the precursor
Synthesis of most lipids requires fatty acids and metabolite
α-ketoglutarate.
glycerol. To produce fatty acids, the acetyl group of
acetyl-CoA (the precursor metabolite produced in
the transition step) is transferred to a carrier protein.
This carrier holds the developing fatty acid chain as NH2
Oxaloacetate
2-carbon units are progressively added. When the
fatty acid reaches its required length, usually 14, 16,
or 18 carbon atoms long, it is released. The glycerol Glutamate
component of the fat is synthesized from the precur-
sor metabolite dihydroxyacetone phosphate, which The amino group (NH2) of glutamate can be
is generated in glycolysis. transferred to other carbon compounds to
produce other amino acids.

Amino Acid Synthesis FIGURE 6.29 The Role of Glutamate in Amino Acid Synthesis Once glutamate
has been synthesized from α-ketoglutarate, its amino group can be transferred to
Proteins are composed of various combinations of usu- produce other amino acids.
ally 20 different amino acids. These amino acids can
be grouped into structurally related families that share ? Alpha-ketoglutarate is produced in which central metabolic pathway?

common pathways of biosynthesis.

Glutamate The pathway begins with the joining of two precursor

Amino acids are necessary for protein synthesis, but glutamate is espe- metabolites—phosphoenolpyruvate (3-carbon) and erythrose
cially important because its synthesis provides a mechanism for bacte- 4-phosphate (4-carbon)—to form a 7-carbon compound. The pre-
ria to incorporate nitrogen into organic material. Recall from chapter 4 cursors originate in glycolysis and the pentose phosphate pathway,
that many bacteria use ammonium (NH4+) as their source of nitrogen; respectively. Then, the 7-carbon compound is modified through a
it is primarily through the synthesis of glutamate that they do this. series of steps until a branch point is reached. At this juncture, two
Glutamate is synthesized in a single-step reaction that adds options are possible. If synthesis proceeds in one direction, tryp-
ammonia to the precursor metabolite α-ketoglutarate, produced in tophan is produced. In the other direction, another branch point is
the TCA cycle (figure 6.29). Once glutamate has been formed, its reached; from there, either tyrosine or phenylalanine can be made.
amino group can be transferred to other carbon compounds to When an amino acid is provided to a cell, it would be a waste
produce amino acids such as aspartate. This transfer of the amino of carbon, energy, and reducing power for that cell to continue
group, a transamination, regenerates α-ketoglutarate from gluta- synthesizing it. But with branched pathways, how does the cell
mate. The α-ketoglutarate can then be used again to incorporate stop synthesizing the product of only one branch? In the pathway
more ammonia. amino group, p. 25 for aromatic acid biosynthesis, this partly occurs by regulating the
enzymes at the branch points. Tryptophan is a feedback inhibitor
Aromatic Amino Acids of the enzyme that directs the branch to its synthesis; this sends the
Synthesis of aromatic amino acids such as tyrosine, phenylala- pathway to the steps leading to the synthesis of the other amino
nine, and tryptophan requires a multistep, branching pathway acids—tyrosine and phenylalanine. Likewise, these two amino
(figure  6.30). This serves as an excellent illustration of many acids each inhibit the first enzyme of the branch leading to their
important features of the regulation of amino acid synthesis. synthesis.
From glycolysis
Phenylalanine

Compound Branch
3-C a point II
7-C Branch
+ Tyrosine
compound point I
4-C
Compound Tryptophan
b
From pentose
phosphate pathway
FIGURE 6.30 Synthesis of Aromatic Amino Acids A multistep branching pathway is used to synthesize aromatic amino acids. The end
product of a branch inhibits the first enzyme of that branch; that end product also inhibits one of the three enzymes that catalyze the first step of
the pathway.
? The synthesis of aromatic amino acids requires precursor metabolites made in which central metabolic pathways?
 Part I Life and Death of Microorganisms 157

In addition to regulating the branchpoints, the three amino Then, in a highly ordered sequence, atoms from the other sources
acids each control the first step of the pathway, the formation of are added to form the purine ring. This can then be converted to a
the 7-carbon compound. In E. coli, three different enzymes can purine nucleotide. To synthesize pyrimidine nucleotides, the pyrim-
catalyze this step; each has the same active site, but they have dif- idine ring is made first and then attached to ribose 5-phosphate.
ferent allosteric sites. Each aromatic amino acid acts as a feedback After one pyrimidine nucleotide is formed, the nucleobase compo-
inhibitor for one of the enzymes. If all three amino acids are pres- nent can be converted into any of the other pyrimidines.
ent in the environment, then very little of the 7-carbon compound
will be synthesized. If only one or two of those amino acids are MicroAssessment 6.10
present, then proportionally more of the compound will be synthe- Biosynthetic processes of different organisms are remarkably similar,
sized. allosteric enzymes, p. 137 using precursor metabolites, NADPH, and ATP to form subunits.
Synthesis of the amino acid glutamate provides a mechanism for
Nucleotide Synthesis bacteria to incorporate nitrogen in the form of ammonia into organic
Nucleotide subunits of DNA and RNA are composed of three material. Synthesis of aromatic amino acids involves branching
pathways. The purine nucleotides are synthesized in a very different
units: a 5-carbon sugar, a phosphate group, and a nucleobase,
manner from the pyrimidine nucleotides.
either a purine or a pyrimidine. They are synthesized as ribonucle-
otides, but these can then be converted to deoxyribonucleotides by 26. Explain why glutamate synthesis is particularly important
for a cell.
replacing the hydroxyl group on the 2′ carbon of the sugar with a
hydrogen atom. nucleotides, p. 32 purine, p. 32 pyrimidine, p. 32 27. What three general products of the central metabolic pathways
does a cell require to carry out biosynthesis?
The purine (double-ring structure) and pyrimidine (single ring)
nucleotides are synthesized in distinctly different manners. The 28. With a branched biochemical pathway, why would it be
important for a cell to shut down the first step as well as
starting compound of purine synthesis is ribose 5-phosphate, a pre- branching steps? +
cursor metabolite generated in the pentose phosphate pathway.

FUTURE CHALLENGES 6.1

Fueling the Future
Considering that microbes are masters at collectively referred to as advanced biofuels. with enough sunlight to support their
extracting energy from seemingly unlikely These include: abundant growth generally lack sufficient
sources, it should not be surprising that they ■ Bioethanol. Rather than using edible water for the final steps of fuel production.
are being enlisted to help us meet our future foods, bioethanol produced with advanced Another option being explored for biodiesel
energy needs. Bacteria, yeast, and algae play technologies is made from cornstalks or production is to genetically engineer an easy-
starring roles in most of the current research other plant waste materials. One problem to-grow bacterium such as E. coli to become
efforts toward developing cost-effective is that the principle component of plant a biodiesel-producing factory, which a lab
methods to produce biofuels, which are fuels wastes is cellulose, and relatively few has recently done. The strain has also been
made from renewable resources. One goal microbes degrade this carbohydrate. engineered to produce the enzyme that
in producing and using these is to be carbon Microbes that degrade cellulose generally breaks down cellulose, so it can use waste
neutral, meaning that fuel production con- are not as easy to grow on a large-scale materials as an energy source. The efficiency
sumes as much atmospheric carbon as fuel basis as yeast, so technical hurdles must is still low, however, so improvements are
use releases. be overcome for this to be a feasible still necessary. Also, the engineered strain
First-generation biofuels are made option. still relies on a second organism—in this
using standard technologies (such as fer- ■ Biodiesel. Composed of fatty acid methyl case, plants—to convert radiant energy into
mentations) and readily available mate- esters, biodiesel has an advantage over chemical energy.
rial (including corn and sugar cane). One bioethanol in that it has a higher energy ■ Biohydrogen. Some cyanobacteria and
example is using yeast to ferment the sugars content and is more compatible with current anoxygenic phototrophs make hydrogen
in corn to produce bioethanol. Although fuel storage and distribution systems. Algae gas as a by-product of nitrogen fixation
this provides an alternative to fossil fuels, and cyanobacteria—both of which are and grow using little more than sunlight
it uses an edible resource, contributing to photosynthetic—capture radiant energy and water. An advantage of biohydrogen
food shortages and increased food prices. In to make lipids that can be used to produce is that it can be easily captured from
addition, the farming practices used to grow biodiesel. This provides a particularly a culture. As with the other advanced
corn on a large-scale basis can adversely attractive option because it bypasses the biofuels, however, there are still technical
effect the environment, and the process for need for the two-stage process used to obstacles that must be overcome before
making bioethanol uses fossil fuels so the create bioethanol (plants convert sunlight to the gas can be produced and stored on a
product is not carbon neutral. chemical energy, and then microbes ferment large-scale basis.
In response to the problems with first- the plant material to produce ethanol). A Will microbes end our reliance on fossil
generation biofuels, newer technologies problem with using photosynthetic microbes fuels? Only time will tell, but the possibilities
are being developed to produce what are to produce biofuels, however, is that areas are exciting!
158 Chapter 6 Metabolism: Fueling Cell Growth

Summary
6.1 ■ Principles of Metabolism 6.3 ■ The Central Metabolic Pathways (table 6.6)
Catabolism is the set of processes that capture and store energy
Glycolysis (figure 6.16)
by breaking down complex molecules. Anabolism includes processes
that use energy to make and assemble the building blocks of a cell Glycolysis converts one molecule of glucose into two molecules of
(figure 6.1). pyruvate; the theoretical net yield of the pathway is 2 ATP, 2 NADH +
H+. Six different precursor metabolites are produced.
Harvesting Energy
Pentose Phosphate Pathway
Photosynthetic organisms harvest the energy of sunlight, using it to
power the synthesis of organic compounds. Chemoorganotrophs The pentose phosphate pathway forms NADPH + 6 H+ and two differ-
harvest energy contained in organic compounds (figure 6.3). Exergonic ent precursor metabolites.
reactions release energy; endergonic reactions use energy. Transition Step (figure 6.17)
Components of Metabolic Pathways The transition step converts pyruvate to acetyl-CoA. Repeated twice, this
A specific enzyme facilitates each step of a metabolic pathway produces 2 NADH + 2 H+. The end product is a precursor metabolite.
(figure 6.5). ATP is the energy currency of the cell. The energy source Tricarboxylic Acid (TCA) Cycle (figure 6.17)
is oxidized to release its energy (figure 6.7). The redox reactions reduce
The TCA cycle completes the oxidation of glucose; the theoretical yield
an electron carrier (figure  6.8). NAD+/NADH, NADP+/NADPH. and
of two “turns” is 6 NADH + 6 H+, and 2 FADH2, in addition to 2 ATP.
FAD/FADH2 are electron carriers (table 6.1).
Two intermediates are precursor metabolites.
Precursor Metabolites
Precursor metabolites are used to make the subunits of macromol- 6.4 ■ Respiration
ecules, and they can also be oxidized to generate energy in the form of
The Electron Transport Chain—Generating Proton
ATP (table 6.2).
Motive Force
Overview of Catabolism (figure 6.10) The electron transport chain sequentially passes electrons, and, as
The central metabolic pathways are glycolysis, the pentose phos- a result, ejects protons. In the mitochondrial electron transport chain,
phate pathway, and the tricarboxylic acid cycle (TCA cycle). three different complexes (complexes I, III, and IV) function as proton
Respiration uses the reducing power accumulated in the central pumps (figure  6.19). Prokaryotes vary with respect to the types and
metabolic pathways to generate ATP by oxidative phosphorylation. arrangements of their electron transport components (figure 6.20). Some
Aerobic respiration uses O2 as a terminal electron acceptor; anaerobic prokaryotes can use molecules other than O2 as terminal electron accep-
respiration uses a molecule other than O2 as a terminal electron accep- tors. This process of anaerobic respiration harvests less energy than
tor (table 6.3). Fermentation uses pyruvate or a derivative as a terminal aerobic respiration.
electron acceptor; this recycles the reduced electron carrier NADH.
ATP Synthase—Harvesting the Proton Motive Force
to Synthesize ATP
6.2 ■ Enzymes
Enzymes function as biological catalysts; they are neither consumed nor ATP synthase permits protons to flow back across the membrane, har-
permanently changed during a reaction. vesting the energy released to fuel the synthesis of ATP.

Mechanisms and Consequences of Enzyme Action (figure 6.11) ATP Yield of Aerobic Respiration in Prokaryotes (figure 6.21)
The substrate binds to the active site, forming an enzyme-substrate The theoretical maximum yield of ATP of aerobic respiration is
complex that lowers the activation energy of the reaction. 38 ATP.

Cofactors (figure 6.12; table 6.4) 6.5 ■ Fermentation

Enzymes sometimes act in conjunction with cofactors such as coen- In general, the only ATP-yielding reactions of fermentations are those
zymes and trace elements. of the glycolytic pathway; the other steps provide a mechanism for
Environmental Factors That Influence Enzyme recycling NADH (figure  6.22). Some end products of fermentation are
Activity (figure 6.13) commercially valuable (figure 6.23). Certain end products help in bacte-
The factors most important in influencing enzyme activities are tem- rial identification.
perature, pH, and salt concentration.
6.6 ■
Catabolism of Organic Compounds
Allosteric Regulation (figure 6.14) Other Than Glucose (figure 6.24)
Cells can fine-tune the activity of an allosteric enzyme by using a regu- Hydrolytic enzymes break down macromolecules into their respective
latory molecule that binds to the allosteric site of the enzyme. subunits.
Enzyme Inhibition Polysaccharides and Disaccharides
Non-competitive inhibition occurs when the inhibitor and the substrate Amylases digest starch, releasing glucose subunits, and are produced by
act at different sites on the enzyme. Competitive inhibition occurs many organisms. Cellulases degrade cellulose. Sugar subunits released
when the inhibitor competes with the normal substrate for the active when polysaccharides are broken down can then enter glycolysis to be
binding site (figure 6.15). oxidized to pyruvate.
 Part I Life and Death of Microorganisms 159

Lipids by ATP synthase to fuel the synthesis of ATP. Photosystems I and II

Fats are hydrolyzed by lipases, releasing glycerol and fatty acids. of cyanobacteria and chloroplasts raise the energy level of electrons
Glycerol is converted to dihydroxyacetone phosphate; fatty acids are stripped from water to a high enough level to be used to generate a pro-
degraded by β-oxidation, generating reducing power and the precursor ton motive force and produce reducing power; this process is oxygenic
metabolite acetyl-CoA. (figure  6.26). Purple and green bacteria use only a single photosystem;
they must obtain electrons from a reduced compound other than water
Proteins and therefore do not generate oxygen.
Proteins are hydrolyzed by proteases. Deamination removes the amino
group; the remaining carbon skeleton is then converted into the appro- 6.9 ■ Carbon Fixation
priate precursor molecule.
Calvin Cycle (figure 6.27)
6.7 ■ Chemolithotrophs The most common pathway used to incorporate CO2 into an organic
Prokaryotes, as a group, are unique in their ability to use reduced inor- form is the Calvin cycle.
ganic compounds such as hydrogen sulfide (H2S) and ammonia (NH3)
as a source of energy. Chemolithotrophs are autotrophs. 6.10 ■ Anabolic Pathways—Synthesizing Subunits from
Precursor Molecules (figure 6.28)
6.8 ■ Photosynthesis
The light reactions capture energy from light and convert it to Lipid Synthesis
chemical energy in the form of ATP. That energy is used for carbon The fatty acid components of fat are synthesized by progressively
fixation. adding 2-carbon units to an acetyl group. The glycerol component is
synthesized from dihydroxyacetone phosphate.
Capturing Radiant Energy
Various pigments such as chlorophylls, bacteriochlorophylls, carot- Amino Acid Synthesis
enoids, and phycobilins are used to capture radiant energy. Reaction Synthesis of glutamate from α-ketoglutarate and ammonia provides
center pigments function as the electron donor in the photosynthetic a mechanism for cells to incorporate nitrogen into organic molecules
process; antennae pigments funnel radiant energy to the reaction (figure  6.29). Synthesis of aromatic amino acids requires a multistep
center pigment. branching pathway. Allosteric enzymes regulate key steps of the path-
way (figure 6.30).
Converting Radiant Energy into Chemical Energy
The high-energy electrons emitted by reaction center chlorophylls Nucleotide Synthesis
are passed to an electron transport chain, which uses them to generate Purines and pyrimidine nucleotides are made in distinctly different
a proton motive force. The energy of proton motive force is harvested manners.

Review Questions
Short Answer 2. Which of the following statements is false? Enzymes
1. Explain the difference between catabolism and anabolism. a) bind to substrates.
2. How does ATP serve as a carrier of free energy? b) lower the energy of activation.
3. How do enzymes catalyze chemical reactions? c) convert coenzymes to products.
d) speed up biochemical reactions.
4. Explain how precursor molecules serve as junctions between cata-
bolic and anabolic pathways. e) can be named after the kinds of reaction they catalyze.
5. How do cells regulate enzyme activity? 3. Which of these is not a coenzyme?
6. Why do the electrons carried by FADH2 result in less ATP produc- a) FAD b) Coenzyme A c) NAD+ d) ATP e) NADP+
tion than those carried by NADH? 4. What is the end product of glycolysis?
7. Name three food products produced with the aid of microorgan- a) Glucose b) Citrate c) Oxaloacetate
isms. d) α-ketoglutarate e) Pyruvate
8. In photosynthesis, what is encompassed by the term “light- 5. The major pathway(s) of central metabolism are
independent reactions”? a) glycolysis and the TCA cycle only.
9. Unlike the cyanobacteria, the anoxygenic photosynthetic bacteria b) glycolysis, the TCA cycle, and the pentose phosphate
do not produce O2. Why not? pathway.
10. What is the role of transamination in amino acid biosynthesis? c) glycolysis only.
d) glycolysis and the pentose phosphate pathway only.
Multiple Choice e) the TCA cycle only.
1. Which of these factors does not affect enzyme activity?
a) Temperature b) Inhibitors c) Coenzymes
d) Humidity e) pH
160 Chapter 6 Metabolism: Fueling Cell Growth

6. Which of these pathways gives a cell the potential to produce the Applications
most ATP? 1. A worker in a cheese-making facility argues that whey, a nutrient-
a) TCA cycle rich by-product of cheese, should be dumped in a nearby pond
b) Pentose phosphate pathway where it could serve as fish food. Explain why this proposed action
c) Lactic acid fermentation could actually kill the fish by depleting the O2 in the pond.
d) Glycolysis 2. Scientists working with DNA in vitro often store it in solutions that
7. In fermentation, the terminal electron acceptor is contain EDTA, a chelating agent that binds magnesium (Mg2+).
a) oxygen (O2). b) hydrogen (H2). This is done to prevent enzymes called DNases from degrading the
DNA. Explain why EDTA would interfere with enzyme activity.
c) carbon dioxide (CO2). d) an organic compound.
8. In the process of oxidative phosphorylation, the energy of proton
motive force is used to generate
Critical Thinking +
a) NADH. b) ADP. c) ethanol. d) ATP. e) glucose. 1. A student argued that aerobic and anaerobic respiration should
produce the same amount of ATP. He reasoned that they both use
9. In the TCA cycle, the carbon atoms contained in acetate are con-
basically the same process; only the terminal electron acceptor is
verted into
different. What is the primary error in this student’s argument?
a) lactic acid. b) glucose. c) glycerol.
2. Chemolithotrophs near hydrothermal vents support a variety of
d) CO2. e) all of these. other life-forms there. Explain how their role is analogous to that
10. Degradation of fats as an energy source involves all of the of photosynthetic organisms in terrestrial environments.
following except
a) β-oxidation. b) acetyl-CoA. c) glycerol.
d) lipase. e) transamination.
7
The Blueprint of Life,
from DNA to Protein
KEY TERMS
Codon A series of three nucleotides Ribosome Structure that facilitates
that code for a specific amino acid. the joining of amino acids during
translation; it is composed of
DNA Polymerase Enzyme that
ribosomal RNA (rRNA) and protein.
synthesizes DNA, using an existing
strand as a template to make the RNA Polymerase Enzyme that
complementary strand. synthesizes RNA using one strand of
DNA as a template.
DNA Replication Duplication of a
DNA molecule. Transcription The process that
copies the information encoded by
Gene The functional unit of the
DNA into RNA.
genome; it encodes a product, most
often a protein. Transfer RNA (tRNA) Type of
RNA molecule that acts as a key,
Genome Complete set of genetic
interpreting the genetic code; each
information in a cell or a virus.
tRNA molecule carries a specific
Messenger RNA (mRNA) Type amino acid.
of RNA molecule translated during
Translation The process that uses
protein synthesis.
the information carried by mRNA to
Promoter Nucleotide sequence synthesize the encoded protein.
to which RNA polymerase binds to
start transcription.
Ribosomal RNA (rRNA) Type of
Model of DNA double helix.
RNA molecule present in ribosomes.

A Glimpse of History
are enzymes. In 1958, Beadle and Tatum were awarded a Nobel Prize,
In 1866, the Czech-Austrian monk Gregor Mendel determined that traits
largely for these pioneering studies that ushered in the era of modern biol-
are inherited as physical units, now called genes. The precise function of
ogy. polypeptide, p. 27
genes, however, was not revealed until 1941, when George Beadle and
Edward Tatum published a scientific paper reporting that genes direct the
production of enzymes.

C
 
onsider for a moment the incredible diversity of life-forms
Beadle and Tatum set out to discover how genes control metabolic in our world—from the remarkable variety of microorgan-
reactions by studying common bread molds that have very simple nutri-
isms, to the plants and animals consisting of many differ-
tional requirements. These molds, Neurospora species, can grow on
media containing only sucrose, inorganic salts, and biotin. Beadle and
ent specialized cells. Every characteristic of each of these cells,
Tatum reasoned that if they created mutant strains which required addi- from its shape to its function, is dictated by information within its
tional nutrients, they could use them to gain insights into the relationship deoxyribonucleic acid (DNA). DNA encodes the master plan, the
between genes and enzymes. For example, a mutant that requires a certain blueprint, for all of an organism’s features.
amino acid likely has a defect in an enzyme required to synthesize that DNA itself is a simple structure, a string composed of only
amino acid. four different nucleotides, each containing a particular nucleobase
To generate mutant strains, Beadle and Tatum treated the mold (also simply called a base): adenine (A), thymine (T), cytosine
cultures with X rays and then grew the resulting cells on a nutrient- (C), or guanine (G). A set of three nucleotides encodes a specific
rich medium that supported the growth of both the original strain and amino acid; in turn, a string of amino acids makes up a protein,
any mutants. Next, they screened thousands of the progeny to find the the structure and function of which is dictated by the order of the
nutrient-requiring mutants. To identify the metabolic defect of each one,
amino acid subunits. Some proteins serve as structural compo-
they grew them separately in various types of media containing different
nutrients.
nents of a cell. Others, such as enzymes, direct cellular activities
Eventually, Beadle and Tatum established that the metabolic defect including biosynthesis and energy conversion. Together, all these
was inherited as a single gene, which ultimately led to their conclusion proteins control the cell’s structure and activities, dictating the
that a single gene determines the production of one enzyme. That assump- overall characteristics of that cell.  deoxyribonucleic acid (DNA), p. 32
tion has been modified somewhat, because we now know that some   nucleotide, p.  32   nucleobase, p.  32   amino acid, p.  25   enzymes,
enzymes are made up of more than one polypeptide, and not all proteins p. 135

161
 Focus Figure

Gene Expression

DNA

Transcription
Copies the information in DNA
DNA Replication into RNA.
Duplicates the DNA molecule
so its encoded information
can be passed on to the next
generation.
RNA
Translation
Interprets the information carried
by RNA to synthesize the
encoded protein.

Protein
FIGURE 7.1 Overview of Replication, Transcription, and Translation
? Which is more stable in a cell—DNA or RNA?

While at first it might seem unlikely that the vast array of analysis of the nucleotide sequence of DNA is called genomics.
life-forms could be encoded by a molecule consisting of only four  chromosome, p. 66  plasmid, p. 66
different units (the nucleotides), think about how much informa- All cells must accomplish two general tasks in order to mul-
tion can be transmitted by binary code, the language of all com- tiply. First, the double-stranded DNA must be duplicated before
puters. Using only two units, a simple series of ones and zeros, cell division so that its encoded information can be passed on to
binary can code for each letter of the alphabet. String enough of the next generation (figure 7.1). This is the process of DNA
these together in the right sequence and the letters become words. replication. Second, the information encoded by the DNA must
With longer and longer strings, the words can become complete be decoded so that the cell can synthesize the necessary gene
sentences, chapters, books, or even whole libraries. products. This process, gene expression, involves two interre-
This chapter will focus on the processes bacteria use to repli- lated events—transcription and translation. Transcription cop-
cate their DNA and convert the encoded information into proteins. ies the information encoded in DNA into a slightly different
The mechanisms used by eukaryotic cells have many similarities, molecule, RNA. The RNA serves as a transitional, temporary
but are considerably more complicated, and will only be discussed form of the information and is the one actually deciphered.
briefly. The processes in archaea are sometimes similar to those of Translation interprets information carried by RNA to synthe-
bacteria, but often resemble those of eukaryotic cells. size the encoded protein. The flow of information from
DNA→RNA→protein is often referred to as the central
dogma of molecular biology.
7.1 ■ Overview
Learning Outcomes Characteristics of DNA
1. Compare and contrast the characteristics of DNA and RNA. DNA is usually a double-stranded, helical structure (figure 7.2).
2. Explain why a cell must be able to regulate gene expression. Each strand is composed of a chain of deoxyribonucleotide sub-
units, more commonly called nucleotides. These subunits are
The complete set of genetic information of a cell is referred to as joined together by a covalent bond between the 5PO4 (5 prime
its genome. Technically, this includes plasmids as well as the phosphate) of one nucleotide and the 3OH (3 prime hydroxyl) of
chromosome; however, the term “genome” is often used inter- the next. The designations 5 and 3 refer to the numbered carbon
changeably with chromosome. The genome of all cells is com- atoms of the pentose sugar of the nucleotide (see figure 2.20).
posed of DNA, but some viruses have an RNA genome. The Joining of the nucleotides in this manner creates a series of alter-
functional unit of the genome is a gene. A gene encodes a product nating sugar and phosphate units, called the sugar-phosphate
(called the gene product), most commonly a protein. The study and backbone. Because of the chemical structure of nucleotides and
 Part I Life and Death of Microorganisms 163

5' phosphate 5' end 3' end 3' hydroxyl

HO

Base
pairs Sugar
P O
A T P
Sugar O

Sugar
P O
G C P
Sugar O

Sugar
DNA P O
C G P
Sugar O

Sugar
P O Nucleotide
T A P
Sugar O

Sugar
P O
C G P
Sugar O
Hydrogen
bonds
HO

3' hydroxyl 3' end 5' end 5' phosphate

FIGURE 7.2 The Structure of DNA The two strands in the double helix are complementary. Three hydrogen bonds form between a
G-C base pair and two between an A-T base pair. The strands are antiparallel; one is oriented in the 5 to 3 direction, and its complement
is oriented in the 3 to 5 direction.
? If a 100 base-pair double-stranded DNA fragment has 40 cytosines, how many adenines does it contain?

how they are joined to each other, a single strand of DNA will strands. Separating the two strands is called melting, or denaturing.
always have a 5PO4 at one end and a 3OH at the other. These hydrogen bonds, p. 21
ends are often referred to as the 5 end (5 prime end) and the
3 end (3 prime end).
The two strands of DNA are complementary and are held Characteristics of RNA
together by hydrogen bonds between the nucleobases. Wherever RNA is similar to DNA in many ways, with a few important
an adenine (A) is in one strand, a thymine (T) is in the other; these exceptions. One difference is that RNA is made up of ribonucleo-
opposing A-T bases are held together by two hydrogen bonds. tides rather than deoxyribonucleotides, although in both cases
Similarly, wherever a guanine (G) is in one strand, a cytosine (C) these are usually referred to simply as nucleotides. Another dis-
is in the other. These G-C bases are held together by three hydro- tinction is that RNA contains the nucleobase uracil in place of the
gen bonds, a slightly stronger attraction than that of an A-T pair. thymine found in DNA. Like DNA, RNA consists of a chain of
The characteristic bonding of A to T and G to C is called base- nucleotides, but RNA is usually a single-stranded linear molecule
pairing and is a fundamental characteristic of DNA. Because of much shorter than DNA. ribonucleic acid (RNA), p. 33
the rules of base-pairing, one strand can always be used as a tem- RNA is synthesized using a region of one of the two strands
plate for the synthesis of the opposing strand. of DNA as a template. In making the RNA molecule, or
Although the two strands of DNA in the double helix are transcript, the base-pairing rules apply except that uracil, rather
complementary, they are also antiparallel. That is, they are ori- than thymine, pairs with adenine. The interaction of DNA and
ented in opposite directions. One strand is oriented in the 5 to 3 RNA is only temporary, however, and the transcript quickly sepa-
direction and its complement is oriented in the 3 to 5 direction. rates from the template.
The duplex structure of double-stranded DNA is generally Specific chromosomal regions are used as templates for RNA
quite stable because of the numerous hydrogen bonds that occur synthesis, generating numerous distinct transcripts. Either DNA
along its length. Short fragments of DNA have correspondingly strand may serve as the template, but only one of the two strands
fewer hydrogen bonds, so they are easily separated into single is transcribed in a given region.
164 Chapter 7 The Blueprint of Life, from DNA to Protein

FIGURE 7.3 Three Functional Types of Protein-encoding gene rRNA gene tRNA gene
RNA Molecules The different functional
groups of RNA—messenger RNA (mRNA),
ribosomal RNA (rRNA), and transfer RNA DNA Transcription
(tRNA)—are transcribed from different
genes. The mRNA is translated, and the
tRNA and rRNA fold into characteristic
three-dimensional structures that each Messenger RNA (mRNA) Ribosomal RNA (rRNA) Transfer RNA (tRNA)
play a role in protein synthesis.
Translation
? Ribosomal RNA is a component of
ribosomes. What are ribosomes?
Protein

Three different functional types of RNA are required for gene provides cells with an important regulatory mechanism. If tran-
expression, and these are transcribed from different sets of genes scription of a gene is turned “on,” transcripts will continue to be
(figure 7.3). Most genes encode proteins and are transcribed into available for translation. If it is then turned “off,” the number of
messenger RNA (mRNA). The information encrypted in mRNA transcripts will rapidly decline. By simply regulating the synthesis
is deciphered according to the genetic code, which correlates each of mRNA molecules, a cell can quickly change the levels of protein
set of three nucleotides to a particular amino acid. Some genes are production. Some cells have mechanisms to adjust the stability of
never translated into proteins; instead the RNAs themselves are RNA, providing an additional level of control.
the final products. These genes encode either ribosomal RNA
(rRNA) or transfer RNA (tRNA), each of which plays a different MicroAssessment 7.1
but critical role in protein synthesis. Replication is the process of duplicating double-stranded DNA.
Transcription is the process of copying the information encoded
Regulating Gene Expression in DNA into RNA. Translation is the process of interpreting the
information carried by messenger RNA in order to synthesize the
Although a cell’s DNA can encode thousands of different pro- encoded protein.
teins, not all of them are needed at the same time or in equal 1. How does the 5 end of a DNA strand differ from the 3 end?
quantities (figure 7.4). Because of this, cells require mechanisms
2. What are the base-pairing rules?
to regulate the expression of certain genes.
3. If the nucleotide sequence of one strand of DNA is
A fundamental aspect of gene regulation is the instability of
5 ACGTTGCA 3, what is the sequence of the complementary
mRNA. Within minutes of being produced, transcripts are degraded strand? +
by cellular enzymes. Although this might seem wasteful, it actually

Gene A Gene B Gene C

Low levels of gene A No transcription of gene B Continuous transcription of

transcription generates leads to no synthesis of gene C generates many
some transcripts of protein B. transcripts of the gene.
the gene.

Translation of each of
the gene A transcripts
generates some protein A.

Translation of each of the gene C transcripts

generates many molecules of protein C.

FIGURE 7.4 The Level of Gene Expression Can Be Controlled

? How does the fact that mRNA is quickly degraded help a cell
control gene expression?
 Part I Life and Death of Microorganisms 165

7.2 ■ DNA Replication complete copy. The replication process is generally bidirectional,
meaning it proceeds in both directions from the starting point
Learning Outcome (figure 7.5). This allows a chromosome to be replicated in half the
3. Describe the DNA replication process, including its initiation and time it would take if the process were unidirectional. The progres-
the events that occur at the replication fork. sion of bidirectional replication around a circular DNA molecule
creates two advancing forks where DNA synthesis is occurring.
DNA is replicated in order to create a duplicate molecule, so that These regions, called replication forks, ultimately meet at a termi-
the two cells generated during binary fission can each receive one nating site when the process is complete. binary fission, p. 82

Original
strand

DNA Replication

New
strand

Origin of
replication Original
strand

Replication
forks
Site where
replication
New
ends
strand

Replication of chromosomal Bidirectional replication creates

DNA starts at the origin of two advancing forks where DNA
replication and then proceeds synthesis is occurring. The replication forks ultimately
in both directions. meet at a terminating site. DNA replication is semiconservative,
meaning each of the two molecules
(a) created contains one of the original
strands paired with a newly
Original synthesized strand.
double-stranded
molecule

(b) Replication forks Replication forks

FIGURE 7.5 Replication of a Bacterial Chromosome (a) Process of bidirectional replication. (b) Partially replicated chromosome; an
electron micrograph and a diagrammatic depiction.
? What is a replication fork?
166 Chapter 7 The Blueprint of Life, from DNA to Protein

The two DNA molecules created through replication each

Components of DNA Replication
contain one of the original strands paired with a newly synthesized TABLE 7.1
in Bacteria
strand. Because half of the original molecule is conserved in each
molecule, replication is said to be semiconservative. Component Comment

Replisome The complex of enzymes and other proteins that

Initiation of DNA Replication synthesize DNA
To initiate replication of a DNA molecule, specific proteins must DNA gyrase Enzyme that temporarily breaks the strands of
recognize and bind to a distinct DNA sequence called an origin of DNA, relieving the tension caused by unwinding
replication. Prokaryotic chromosomes and plasmids typically the two strands of the DNA helix.
contain only one of these initiating sites. A molecule that lacks this DNA ligase Enzyme that joins two DNA fragments by
forming a covalent bond between the sugar and
sequence will not be replicated. The proteins that bind to the origin
phosphate residues of adjacent nucleotides.
of replication cause localized melting of the double-stranded DNA,
DNA Enzymes that synthesize DNA; they use one
exposing single-stranded regions that can act as templates. Enzymes polymerases strand of DNA as a template to make the
called primases then synthesize short stretches of RNA comple- complementary strand. Nucleotides can
mentary to the exposed templates. These small fragments, called be added only to the 3 end of an existing
primers, are critical in the next steps of replication. fragment—therefore, synthesis always occurs in
the 5 to 3 direction.

The Process of DNA Replication Helicases Enzymes that unwind the DNA helix ahead of
the replication fork.
The process of DNA replication requires the coordinated action of Okazaki Nucleic acid fragment produced during
many different enzymes and other proteins. The most critical of fragment discontinuous synthesis of the lagging strand
these exist together in DNA-synthesizing “assembly lines” called of DNA.
replisomes (table 7.1). Origin of Distinct region of a DNA molecule at which
Enzymes called DNA polymerases synthesize DNA in the replication replication is initiated.
5 to 3 direction, using one strand as a template to make the Primase Enzyme that synthesizes small fragments of RNA
complement (figure 7.6). To do this, DNA polymerase adds to serve as primers for DNA synthesis.
nucleotides onto the 3 end of a primer, powering the reaction Primer Fragment of nucleic acid to which DNA
with the energy released when a high-energy phosphate bond polymerase can add nucleotides (the enzyme can
add nucleotides only to an existing fragment).
of the incoming nucleotide is hydrolyzed. Note that DNA

Template strand
5' 3'

T A C G G T A C T A G T A G T A G T C G A T T C G A A
DNA Replication
T C A T C A T C A G C T A A G C T T

Direction
3' 5'
of synthesis A
DNA polymerase New strand

FIGURE 7.6 The Process of DNA P P P

Synthesis DNA polymerase synthesizes
a new strand by adding one nucleotide
at a time to the 3 end of the elongating O O O
strand. The base-pairing rules determine
the specific nucleotides that are added.
T A G
? Considering that DNA is synthesized
in the 5 to 3 direction, which A T C
direction must DNA polymerase
travel along the template strand:
5 to 3 or 3 to 5? O O O

OH OH
P~
P~ P P
P
 Part I Life and Death of Microorganisms 167

polymerases add nucleotides only onto an existing

DNA strand, so they cannot initiate synthesis. This
explains why primers are required at the origin of rep- Replication forks
lication—they provide the DNA polymerase with a
molecule to which it can add additional nucleotides.
  high-energy phosphate bond, p. 24

In order for replication to progress, enzymes called 3'

helicases must progressively “unzip” the DNA strands at 5'
1 A helicase “unzips”
each replication fork to reveal additional template the two strands of DNA. Helicase
sequences (figure 7.7 1 ). 2 Synthesis of one new strand Leading
proceeds continuously as fresh template is exposed, strand
5'
because DNA polymerase simply adds nucleotides to the 5'
3' RNA primer
3 end. This strand is called the leading strand.
3   Synthesis of the other strand, the lagging strand, is
more complicated. This is because DNA polymerases can- DNA polymerase adds
not add nucleotides to the 5 end, so as additional template nucleotides onto the 3'
end of the strand. 5'
is exposed, synthesis must be reinitiated. Each time syn-
thesis is reinitiated, another RNA primer must be made
first. The result is a series of small fragments, each of 3'
2 Synthesis of the leading strand 5'
which has a short stretch of RNA at its 5 end. These proceeds continuously as fresh
fragments are called Okazaki fragments. 4 As DNA template is exposed.
polymerase adds nucleotides to the 3 end of one Okazaki
fragment, it eventually reaches the 5 end of another. A 5'
different type of DNA polymerase then removes the RNA 3' 5'
primer nucleotides and simultaneously replaces them with Okazaki fragment
of the lagging strand
deoxynucleotides. 5 The enzyme DNA ligase then seals
the gaps between fragments by forming a covalent bond Primase synthesizes
the RNA primer.
between the adjacent nucleotides.
When a circular bacterial chromosome is replicated,
the two replication forks eventually meet at a site opposite 3 Synthesis of the lagging strand must be reinitiated as more
template is exposed. Each time synthesis is reinitiated, 3'
the origin of replication. Two complete DNA molecules a new RNA primer must be made. Discontinuous synthesis 5'
have been produced at this point, and these can be passed generates Okazaki fragments.
on to the two daughter cells.
It takes approximately 40 minutes for the E. coli chro-
mosome to be replicated. How, then, can the organism
have a generation time of only 20 minutes? This can
happen because, under favorable growing conditions, a
cell initiates replication before the preceding round of rep-
lication is complete. In this way, each of the two daughter
cells will get one complete chromosome that has already
started another round of replication. generation time, p. 83 5'
3' 5'
MicroByte 5 DNA ligase seals
the gaps between
Antibacterial medications called fluoroquinolones target the
Okazaki fragments
enzyme DNA gyrase, a component of the bacterial replisome. by forming a covalent
bond between them.

4 As DNA polymerase adds nucleotides

MicroAssessment 7.2 to the 3' end of one Okazaki fragment,
it encounters the 5' end of another. DNA ligase
DNA replication begins at the origin of replication and then A different type of DNA polymerase
proceeds bidirectionally, creating two replication forks. DNA then removes the RNA primer
polymerases synthesize DNA in the 5 to 3 direction, using one nucleotides and simultaneously 3'
strand as a template to generate the complementary strand. replaces them with deoxynucleotides. 5'
4. Why is a primer required for DNA synthesis?
5. How does synthesis of the lagging strand differ from that
of the leading strand? FIGURE 7.7 The Replication Fork This depiction is simplified to
highlight the key differences between synthesis of the leading and lagging
6. Eukaryotic chromosomes often have multiple origins of strands.
replication. Why would this be the case? +
? Synthesis of which strand requires the repeated action of DNA ligase?
168 Chapter 7 The Blueprint of Life, from DNA to Protein

7.3 ■ Gene Expression in Bacteria TABLE 7.2 Components of Transcription

in Bacteria
Learning Outcomes
Component Comment
4. Describe the process of transcription, focusing on the role of RNA
polymerase, sigma (σ) factors, promoters, and terminators. (–) strand Strand of DNA that serves as the template for
5. Describe the process of translation, focusing on the role of mRNA, RNA synthesis; the resulting RNA molecule is
ribosomes, ribosome-binding sites, rRNAs, tRNAs, and codons. complementary to this strand.
(+) strand Strand of DNA complementary to the one that
Recall that gene expression involves two separate but inter- serves as the template for RNA synthesis; the
related processes, transcription and translation. Transcription is nucleotide sequence of the RNA molecule is
the same as this strand, except it has uracil
the process of synthesizing RNA from a DNA template. During rather than thymine.
translation, information encoded by an mRNA transcript is
Promoter Nucleotide sequence to which RNA polymerase
used to synthesize a protein. Note that the term “polypeptide” binds to initiate transcription.
is more accurate here, but the word “protein” is often used in
RNA polymerase Enzyme that synthesizes RNA using single-
this context for simplicity. The distinction between these two stranded DNA as a template; synthesis always
words is subtle—a polypeptide is simply a chain of amino occurs in the 5 to 3 direction.
acids, whereas a protein is a functional molecule made up of Sigma (σ) factor Component of RNA polymerase that recognizes
one or more polypeptides. the promoter regions. A cell can have different
types of σ factors that recognize different
Transcription promoters, allowing the cell to transcribe
specialized sets of genes as needed.
In transcription, the enzyme RNA polymerase synthesizes single-
Terminator Nucleotide sequence at which RNA synthesis
stranded RNA molecules from a DNA template. Nucleotide
stops; the RNA polymerase falls off the DNA
sequences in the DNA direct the polymerase where to start and template and releases the newly synthesized
where to end (figure 7.8). The DNA sequence to which RNA RNA.
polymerase can bind and initiate transcription is called a promoter;
one that stops the process is a terminator. Like DNA polymerase,
RNA polymerase can add nucleotides only to the 3 end of a chain
Initiation of RNA Synthesis
and therefore synthesizes RNA in the 5 to 3 direction. Unlike
DNA polymerase, however, RNA polymerase can start synthesis Transcription is initiated when RNA polymerase binds to a pro-
without a primer. moter (figure 7.10). The binding melts a short stretch of DNA,
The RNA sequence made during transcription is complemen- creating a region of exposed nucleotides that serves as a template
tary and antiparallel to the DNA template (figure 7.9). The DNA for RNA synthesis.
strand that serves as the template for transcription is called the The portion of RNA polymerase that recognizes the promoter
minus (–) strand, and its complement is called the plus (+) is a loosely attached subunit called sigma (σ) factor. A cell can
strand (table 7.2). Because the RNA is complementary to the (–) produce various types of σ factors, each recognizing different pro-
DNA strand, its nucleotide sequence is the same as the (+) DNA moters. By controlling which σ factors are made, cells can tran-
strand, except it contains uracil rather than thymine. scribe specialized sets of genes as needed. The RNA polymerases
In prokaryotes, mRNA molecules can carry the information
for one or multiple genes. A transcript that carries one gene is FIGURE 7.9 RNA Is Complementary
called monocistronic (a cistron is synonymous with a gene). One Gene Expression
and Antiparallel to the DNA
that carries multiple genes is called polycistronic. The proteins Template The DNA strand that serves
Transcription as a template for RNA synthesis is
encoded on a polycistronic message generally have related func-
called the (–) strand of DNA. The
tions, allowing a cell to express related genes as one unit.
complement to that is the (+) strand.
Translation
Region transcribed
? How does the nucleotide sequence
of the (+) strand differ from that
of the RNA transcript?

DNA Promoter Transcription Terminator

5' 3' Plus (+)
G C T G A T G A T C C G C G T A GG T G C T strand
5' 3' of DNA
C G A C T A C T A GG C G C A T C C A C G A
RNA
3' 5' Minus (–)
FIGURE 7.8 Nucleotide Sequences in DNA Direct strand
Transcription The promoter is a DNA sequence to which RNA of DNA
polymerase can bind in order to initiate transcription. The terminator
is a sequence at which transcription stops. 5' 3' RNA
? In which direction is RNA synthesized: 5 to 3 or 3 to 5? G C U G A U G A U C C G C G U A GG U G C U
 Part I Life and Death of Microorganisms 169
Gene Expression

FIGURE 7.10 The Process of RNA Synthesis Bacterial RNA polymerases include a sigma subunit (as
Transcription
illustrated); the RNA polymerases of eukaryotic cells and archaea use transcription factors to recognize promoters.
? Which component of the bacterial RNA polymerase recognizes the promoter?
Translation

5' 3'
3' 5'
Promoter Terminator

RNA polymerase
1 Initiation
5' 3' RNA polymerase binds to the
3' 5' promoter and melts a short
stretch of DNA.
Template
Sigma strand

2 Elongation
Sigma factor dissociates from RNA
5' 3' polymerase, leaving the core
3' 5' enzyme to complete transcription.
RNA is synthesized in the 5' to 3'
Promoter direction as the enzyme adds
5' nucleotides to the 3' end of
RNA
the growing chain.
GA C U G
C T GAC

3 Termination
5' 3' When RNA polymerase
encounters a terminator, it falls
3' 5' off the template and releases
Promoter the newly synthesized RNA.

5'

RNA polymerase
dissociates from template.

of eukaryotic cells and archaea use proteins called transcription which strand will be used as a template (figure 7.11). The direc-
factors to recognize promoters. tion of polymerase movement can be likened to the flow of a river.
Promoters identify the regions of a DNA molecule that will Because of this, the words “upstream” and “downstream” are used
be transcribed into RNA. In doing so, they also orient the direction to describe relative positions of other sequences. As an example,
of the RNA polymerase on the DNA molecule, thereby dictating promoters are upstream of the genes they control.

The orientation of the promoter dictates the

direction of transcription, and this determines
Terminator Template which strand is used as a template. Terminator
strand
5' 3'
DNA
3' 5'
Promoter 1 Promoter 2 Template strand

RNA 3' 5' 5' 3' RNA

FIGURE 7.11 The Promoter Orients RNA Polymerase The orientation of RNA polymerase determines which strand will be used as
the template. In this diagram, the color of the RNA molecules indicates which DNA strand was used as the template. The light blue RNA was
transcribed from the red DNA strand (and is therefore analogous in sequence to the blue DNA strand), whereas the pink RNA was transcribed
from the blue DNA strand (and is therefore analogous in sequence to the red DNA strand).
? The light blue RNA strand is complementary to which DNA strand in this figure? To which DNA strand is the pink RNA strand complementary?
170 Chapter 7 The Blueprint of Life, from DNA to Protein

TABLE 7.3 Components of Translation in Bacteria

Component Comment

Anticodon Sequence of three nucleotides in a tRNA molecule that is complementary to a particular codon in mRNA. The
anticodon allows the tRNA to recognize and bind to the appropriate codon.
mRNA Type of RNA molecule that contains the genetic information deciphered during translation.
Polyribosome Multiple ribosomes attached to a single mRNA molecule.
(polysome)
Reading frame Grouping of a stretch of nucleotides into sequential triplets; an mRNA molecule has three potential reading frames,
but only one is typically used in translation.
Ribosome Structure that facilitates the joining of amino acids during the process of translation; composed of protein and
ribosomal RNA. The prokaryotic ribosome (70S) consists of a 30S and 50S subunit.
Ribosome-binding site Sequence of nucleotides in mRNA to which a ribosome binds; the first time the codon for methionine (AUG) appears
after that site, translation generally begins.
rRNA Type of RNA molecule present in ribosomes.
Start codon Codon at which translation is initiated; it is typically the first AUG after a ribosome-binding site.
Stop codon Codon that terminates translation, signaling the end of the protein; there are three stop codons.
tRNA Type of RNA molecule that acts as a key that interprets the genetic code; each tRNA molecule carries a specific amino
acid.

Elongation of the RNA Transcript The Role of mRNA

In the elongation phase, RNA polymerase moves along The mRNA is a temporary copy of genetic information; it carries
DNA, using the (–) strand as a template to synthesize encoded instructions for synthesis of a specific protein, or in
a single-stranded RNA molecule (see figure 7.10). the case of a polycistronic message, a specific group of
Gene Expression
As with DNA replication, nucleotides are added proteins. That information is deciphered using the
only to the 3 end; the reaction is fueled by Transcription genetic code, which correlates a series of three nucle-
hydrolyzing a high-energy phosphate bond of otides, a codon, with one amino acid (table 7.4). The
the incoming nucleotide. When RNA poly- genetic code is practically universal, meaning that it
merase advances, it denatures a new stretch of Translation is used in nearly its entirety by all living things.
DNA and allows the previous portion to close.
This exposes a new region of the template so elon-
Region translated
gation can continue.
Once elongation has proceeded far enough for RNA 5' 3'
polymerase to clear the promoter, another molecule of the enzyme mRNA
can bind, initiating a new round of transcription. Thus, a single Ribosome- Start Stop
binding site codon Translation codon
gene can be transcribed repeatedly very quickly.

Termination of Transcription
Just as an initiation of transcription occurs at a distinct site on the Protein
DNA, so does termination. When RNA polymerase encounters a
sequence called a terminator, it falls off the DNA template and
releases the newly synthesized RNA.
Ser His
Phe Cys
Tyr Glu Val Gly
Tyr
Ser Ala
Pro Leu
Gln
Ser Met

Translation
FIGURE 7.12 Nucleotide Sequences in mRNA Direct
Translation is the process of decoding the information carried on Translation The ribosome begins to assemble at the ribosome-
the mRNA to synthesize the specified protein. The process binding site and starts translating at the start codon. Translation ends
requires three major structures—mRNA, ribosomes, and tRNAs— at the stop codon.
in addition to various other components (table 7.3). ? In which direction does the ribosome move along RNA?
 Part I Life and Death of Microorganisms 171

TABLE 7.4 The Genetic Code

Second Letter
First Third
Letter U C A G Letter
U UUU UCU UAU Tyr Tyrosine UGU U
Phe Phenylalanine Cys Cysteine
UUC UCC UAC UGC C
Ser Serine
UUA UCA UAA “Stop” UGA “Stop” A
Leu Leucine
UUG UCG UAG “Stop” UGG Trp Trytophan G

C CUU CCU CAU CGU U

His Histidine
CUC CCC CAC CGC C
Leu Leucine Pro Proline Arg Arginine
CUA CCA CAA CGA A
Gln Glutamine
CUG CCG CAG CGG G

A AUU ACU AAU AGU U

Asn Asparagine Ser Serine
AUC Ile Isoleucine ACC AAC AGC C
Thr Threonine
AUA ACA AAA AGA A
Lys Lysine Arg Arginine
AUG Met Methionine; “Start” ACG AAG AGG G

G GUU GCU GAU GGU U

Asp Aspartate
GUC GCC GAC GGC C
Val Valine Ala Alanine Gly Glycine
GUA GCA GAA GGA A
Glu Glutamate
GUG GCG GAG GGG G

A codon consists of three nucleotides read in the sequence shown. For example, ACU codes for threonine. The first letter, A, is in the First Letter column; the second
letter, C, is in the Second Letter column; and the third letter, U, is in the Third Letter column. Many amino acids are specified by more than one codon. For example,
threonine is specified by four codons, which differ only in the third nucleotide (ACU, ACC, ACA, and ACG).

Because a codon is a triplet of any combination of the four Reading frame #1 C U G G C A U U G C C U U A U

nucleotides, there are 64 different codons (43). Three are stop
codons, which will be discussed later. The remaining 61 translate
Leu Ala Leu Pro Tyr
to the 20 different amino acids. This means that more than one
codon can code for a specific amino acid. For example, both ACA
and ACG encode the amino acid threonine. Because of this redun-
dancy, the genetic code is said to be degenerate.
Reading frame #2 C U G G C A U U G C C U U A U
The nucleotide sequence of mRNA indicates where the cod-
ing region begins and ends (figure 7.12). The site at which it
begins is particularly critical because the translation “machinery” Trp His Cys Leu
reads the mRNA in groups of three nucleotides. As a conse-
quence, any given sequence has three possible reading frames,
or ways in which triplets can be grouped (figure 7.13). If transla-
Reading frame #3 C U G G C A U U G C C U U A U
tion begins in the wrong reading frame, a very different, and
generally non-functional, protein would be synthesized.
Gly Ile Ala Leu

The Role of Ribosomes

Ribosomes serve as translation “machines,” structures that FIGURE 7.13 Reading Frames A nucleotide sequence has
string amino acids together. A ribosome does this by aligning three potential reading frames, but only one is typically used for
two amino acids so that a ribosomal enzyme can easily create translation.
a peptide bond between them. peptide bond, p. 27 ? Why is it important that the correct reading frame is used?
172 Chapter 7 The Blueprint of Life, from DNA to Protein

Pro
Ribosomes also locate key punctuation sequences on the
Amino acid
mRNA molecule, such as the point at which protein synthesis
should begin. The ribosome then moves along the mRNA in
the 5 to 3 direction, “presenting” each codon in a sequential
order for deciphering, while maintaining the correct reading
frame. ribosomes, p. 66
Prokaryotic ribosomes are composed of a 30S subunit and
Hydrogen bond
a 50S subunit, each made up of protein and ribosomal RNA
(rRNA) (see figures 3.42 and 10.8); the “S” stands for Svedberg
unit, which is a measure of size. Note that Svedberg units are
not additive, which is why the 70S ribosome can have 30S and
50S subunits. ribosomal subunits, p. 246

tRNA
MicroByte
Several types of antibiotics, including tetracycline and azithromycin,
interfere with the function of the bacterial 70S ribosome.
Anticodon

G G C
C C G The Role of Transfer RNAs
5' 3' The tRNAs are segments of RNA that act as keys to the genetic
mRNA
code (figure 7.14). Each tRNA recognizes and base-pairs with
(a) Codon
certain codons and, in the process, delivers the appropriate amino
acid to that site. This recognition is made possible because each
tRNA has an anticodon—three nucleotides complementary to a
Pro codon in the mRNA. The anticodon of a tRNA molecule dictates
which amino acid the molecule carries.
Once a tRNA molecule has donated its amino acid during
translation, it can be recycled. An enzyme in the cytoplasm recog-
tRNA nizes the tRNA and then attaches the appropriate amino acid.

Initiation of Translation
In prokaryotes, translation begins as the mRNA molecule is still
being synthesized (figure 7.15). Part of the ribosome binds to
a sequence in mRNA called the ribosome-binding site; the
first AUG after that site usually serves as the start codon. The
complete ribosome assembles there, joined by an initiating tRNA
G G C that carries a chemically altered form of the amino acid methio-
(b) Anticodon nine (N-formylmethionine, or f-Met). The position of the first
AUG is critical, as it determines the reading frame used for trans-
FIGURE 7.14 The Structure of Transfer RNA (tRNA) lation of the remainder of that protein. Note that AUG functions
(a) Two-dimensional illustration of tRNA. The anticodon of the tRNA base-
pairs with its complementary codon in the mRNA; by doing so, it delivers as a start codon only when preceded by a ribosome-binding site;
the appropriate amino acid to the site. The amino acid that the tRNA carries at other sites, it simply encodes methionine.
is dictated by the genetic code. The tRNA that recognizes the codon CCG
carries the amino acid proline. (b) Three-dimensional illustration of tRNA.
? A tRNA that has the anticodon GAG carries which amino acid? Elongation of the Polypeptide Chain
The ribosome has two sites to which amino acid–carrying tRNAs
can bind—the P-site and the A-site. At the start of translation, the
initiating tRNA carrying the f-Met occupies the P-site (figure 7.16).
A tRNA that recognizes the next codon on the mRNA then fills
the unoccupied A-site. Once both sites are filled, an enzyme cre-
ates a peptide bond between the two amino acids carried by the
tRNAs. This transfers the amino acid from the initiating tRNA to
the amino acid carried by the incoming tRNA.
 Part I Life and Death of Microorganisms 173

Gene Expression

Transcription

Translation

5' 3'
3' 5'
DNA mRNA strand
Ribosome

5' Polypeptide

Ribosome- Start
binding site codon

FIGURE 7.15 In Prokaryotes, Translation Begins as the mRNA Molecule Is Still Being Synthesized Ribosomes begin translating
the 5 end of mRNA before transcription is complete. More than one ribosome can be translating the same mRNA molecule.
? Why is the position of the first AUG after the ribosome-binding site critical?

After the initiating tRNA has donated its amino acid to the Post-Translational Modification
tRNA in the A-site, the ribosome advances a distance of one Polypeptides must often be modified after they are synthesized in
codon, moving along the mRNA in a 5 to 3 direction. As this order to become functional. For example, some must be folded
happens, the initiating tRNA is released through a region called into a specific three-dimensional structure, a process that requires
the E-site. The remaining tRNA, which now carries both amino the assistance of proteins called chaperones. Polypeptides des-
acids, occupies the P-site. The A-site is transiently empty. A tined for transport through the cytoplasmic membrane also must
tRNA that recognizes the codon in the A-site quickly attaches be modified. These have a signal sequence, a characteristic series
there, and the process repeats. of hydrophobic amino acids at their amino terminal end, which
Once translation of a gene has progressed far enough for the “tags” them for transport. The signal sequence must be removed
ribosome to clear the initiating sequences, another ribosome can by proteins in the membrane.
bind. Thus, at any one time, multiple ribosomes can be translating
a single mRNA molecule (see figure 7.15). This allows maximal
protein synthesis from a single mRNA template. The assembly of
multiple ribosomes attached to a single mRNA molecule is called MicroAssessment 7.3
a polyribosome, or a polysome. Gene expression involves transcription and translation. RNA
polymerase synthesizes RNA in the 5 to 3 direction, using one strand
of DNA as a template. Ribosomes move along the resulting mRNA
in the 5 to 3 direction, synthesizing protein. tRNAs carry specific
Termination of Translation amino acids, thereby acting as keys to the genetic code.
Elongation of the polypeptide terminates when the ribosome 7. How does a promoter dictate which DNA strand is used as the
reaches a stop codon, a codon not recognized by a tRNA. At this template?
point, enzymes free the polypeptide by breaking the covalent bond 8. What is the role of tRNA?
that joins it to the tRNA. The ribosome falls off the mRNA, dis-
9. Could two mRNAs have different nucleotide sequences and yet
sociating into its two component subunits (30S and 50S). The code for the same protein? Explain your answer. +
subunits can then be reused to initiate translation at other sites.
174 Chapter 7 The Blueprint of Life, from DNA to Protein

f-Met

P-site Initiation
The initiating tRNA, carrying the amino
E-site A-site acid f-Met, base-pairs with the start
codon and occupies the P-site.

U A C
A U G C C G U A C G A A G A U U A C U A G G A U
5' 3'

mRNA

f-Met Pro

A tRNA that recognizes the next codon

then fills the unoccupied A-site.

U A C G G C
A U G C C G U A C G A A G A U U A C U A G G A U
5' 3'

Peptide bond
f-Met
Pro

The ribosome catalyzes the joining of the

amino acid carried by the tRNA in the
P-site to the one carried by the tRNA in
the A-site.

U A C G G C
A U G C C G U A C G A A G A U U A C U A G G A U
5' 3'

(a)

FIGURE 7.16 The Process of Translation (a) Initiation. (b) Elongation. (c) Termination.
? What would happen if a tRNA that recognized UAA (the stop codon) were present?
 Ty Part I Life and Death of Microorganisms 175
f-Met
r

Pro

Elongation
The ribosome advances a distance of
P-site one codon. The tRNA that occupied the
P-site exits through the E-site and the
C E-site A-site tRNA that was in the A-site occupies the
A
U P-site. A tRNA that recognizes the next
A codon quickly fills the empty A-site.
U
G
G G C
A U G C C G U A C G A A G A U U A C U A G G A U
5' 3'
Ribosome moves along mRNA.

f-Met
G
lu
Pro

Tyr

The ribosome continues advancing down

C the mRNA in the 5' to 3' direction, moving
G G one codon at a time.
C
U
U
A U G
A U G C C G U A C G A A G A U U A C U A G G A U
5' 3'

(b)

f-Met Pro Tyr

Glu
Asp

Tyr

Termination
A Translation continues until a stop codon
U
C is reached, signaling the end of the
process. No tRNA molecules recognize a
stop codon.

A U G
A U G C C G U A C G A A G A U U A C U A G G A U
5' 3'

The components dissemble, releasing the

newly formed polypeptide.

Pro Tyr
f-Met Glu Tyr
Asp

(c)
176 Chapter 7 The Blueprint of Life, from DNA to Protein

TABLE 7.5 Major Differences Between Prokaryotic and Eukaryotic Transcription and Translation
Prokaryotes Eukaryotes

mRNA is not processed. A cap is added to the 5 end of mRNA, and a poly A tail is added to the
3 end.

mRNA does not contain introns. mRNA contains introns, which are removed by splicing.

Translation of mRNA begins as it is being transcribed. The mRNA transcript is transported out of the nucleus so that it can be
translated in the cytoplasm.

mRNA is often polycistronic; translation usually begins at the first mRNA is monocistronic; translation begins at the first AUG.
AUG codon that follows a ribosome-binding site.

7.4 ■ Differences inactivate bacterial 70S ribosomes, but not 80S ribosomes. This
explains why those antibiotics kill bacteria without causing sig-
Between Eukaryotic nificant harm to mammalian cells.
and Prokaryotic
Gene Expression MicroByte
Symptoms of the disease diphtheria are due to a toxin that inactivates
Learning Outcome a protein required for 80S ribosome function.
6. Describe four differences between prokaryotic and eukaryotic gene
expression.

Eukaryotes differ significantly from prokaryotes in several Eukaryotic DNA contains

aspects of transcription and translation (table 7.5). Eukaryotic introns, which interrupt
coding regions (exons).
mRNA, for example, is synthesized in a precursor form,
called pre-mRNA. The pre-mRNA must be processed
(altered), both during and after transcription to form mature Exon Intron Exon Intron Exon
mRNA. Shortly after transcription begins, the 5 end of the
pre-mRNA is capped by adding a methylated guanine deriv- Eukaryotic DNA
ative. This cap binds specific proteins that stabilize the tran- Transcription generates
script and enhance translation. The 3 end of the molecule is pre-mRNA (precursor mRNA)
also modified, even before transcription has been terminated. that contains introns. A cap
and poly A tail are then added.
This process, polyadenylation, cleaves the transcript at a
specific sequence and then adds about 200 adenine deriva- Cap Poly A tail
tives to the new 3 end. This creates a poly A tail, which is
thought to stabilize the transcript as well as enhance transla-
tion. Another important modification is splicing, which Pre-mRNA
removes specific segments of the transcript (figure 7.17).
Splicing is necessary because eukaryotic genes are often
interrupted by non-coding sequences. These intervening
sequences, introns, are transcribed along with the expressed Splicing removes introns to
regions, exons, and must be removed from pre-mRNA to create functional mRNA.
create functional mRNA.
The mRNA in eukaryotic cells must be transported out of the
nucleus before it can be translated in the cytoplasm. Thus, unlike in mRNA
prokaryotes, the same mRNA molecule cannot be synthesized and
mRNA is transported out of
translated at the same time or even in the same cellular location. The the nucleus to be translated
mRNA of eukaryotes is generally monocistronic, and translation of in the cytoplasm.
the message typically begins at the first AUG in the molecule.
The ribosomes of eukaryotes differ from those of prokary-
otes. Whereas the prokaryotic ribosome is 70S, made up of 30S
and 50S subunits, the eukaryotic ribosome is 80S, made up of
40S and 60S subunits. The differences in ribosome structure FIGURE 7.17 Splicing of Eukaryotic RNA
are medically important because certain antibiotics bind to and ? Introns are intervening sequences; what are exons?
 Part I Life and Death of Microorganisms 177

PERSPECTIVE 7.1
RNA: The First Macromolecule?
The 1989 Nobel Prize in Chemistry was proteins had been added, fully expecting that widespread. Ribozymes have been found in
awarded to two Americans, Sidney Altman nothing would happen. Much to his surprise, the mitochondria of eukaryotic cells and
and Thomas Cech, who independently made the introns were removed in the control as shown to catalyze other reactions that resem-
the unexpected observation that RNA mol- well. Based on these results, Cech could only ble the polymerization of RNA.
ecules can act as enzymes. Before their stud- conclude that the RNA acted on itself to cut These observations have profound impli-
ies, it was believed that only proteins had out pieces of RNA. cations for a long-standing question in evolu-
enzymatic activity. The studies of Altman and his col- tionary biology: Which came first, proteins or
Cech made a key observation in 1982 when leagues—carried out simultaneously to, and nucleic acids? The answer seems to be nucleic
he was trying to understand how introns were independently of, Cech’s—showed that RNA acids, specifically RNA, which acted both as
removed from ribosomal pre-RNA (pre-rRNA) had catalytic properties beyond cutting out a carrier of genetic information as well as an
in a eukaryotic protozoan. Because he was con- introns from pre-rRNA. Altman’s group enzyme. Billions of years ago, before the pres-
vinced that proteins were responsible for cutting found that RNA could convert a tRNA mol- ent universe in which DNA, RNA, and protein
out introns, he added all of the protein in the ecule from a precursor form to its final func- are found, the only macromolecule was prob-
cells’ nuclei to a suspension of pre-rRNA. As tional state. Additional studies have shown ably RNA. Once tRNAs became available,
expected, the introns were cut out. For a con- that enzymatic reactions in which catalytic they carried amino acids to nucleotide
trol, Cech used pre-rRNA to which no nuclear RNAs, or ribozymes, play a role are very sequences on a strand of RNA.

however, follows the feast. Between meals—a period of time

MicroAssessment 7.4
that can be many days in the case of some mammals—the rich
Eukaryotic pre-mRNA must be processed, which involves capping, source of nutrients is depleted. Now the bacterial cells’ biosyn-
polyadenylation, and splicing. In eukaryotic cells, the mRNA must thetic pathways must be activated, using energy and slowing cell
be transported out of the nucleus before it can be translated in the
cytoplasm. Eukaryotic mRNA is monocistronic.
growth. Cells dividing several times an hour in a nutrient-rich
environment might divide only once every 24 hours in a starved
10. What is an intron?
mammalian gut. Later, when the animal defecates, some of the
11. Would a deletion of two base pairs have a greater E. coli cells are excreted in the feces. Outside of the mammalian
consequence if it occurred in an intron or in an exon? +
host, the bacterial cells must cope with yet a completely different
set of conditions to survive.

Signal Transduction
7.5 ■ Sensing and Responding to Signal transduction transmits information from outside a cell to
Environmental Fluctuations the inside. This allows cells to monitor and react to environmental
conditions.
Learning Outcomes
7. Describe how quorum sensing and two-component regulatory
systems allow cells to adapt to fluctuating conditions. Quorum Sensing
8. Compare and contrast antigenic variation and phase variation. Some organisms can “sense” the density of cells within their own
population—a phenomenon called quorum sensing. This allows
Microorganisms are constantly faced with rapidly changing cells to activate genes that are only useful when expressed by a
environmental conditions, and must quickly adapt to the fluctua- critical mass. As an example, the cooperative activities leading to
tions if they are to survive. Consider the situation of E. coli in biofilm formation are controlled by quorum sensing. Some patho-
the intestinal tract of mammals. In this habitat, it must cope with gens use the mechanism to coordinate expression of genes involved
alternating periods of feast and famine. For a limited time after with the infection process. biofilms, p. 84
a mammal eats, the bacterial cells prosper, bathed in the mixture Quorum sensing involves a process that allows bacteria to
of amino acids, vitamins, and other nutrients. The cells actively “talk” to each other by synthesizing one or more varieties of extra-
take up these compounds they would otherwise need to synthe- cellular signaling molecules. When few cells are present, the
size. Simultaneously, the cells shut down their biosynthetic concentration of a given signaling molecule is very low. As the
pathways, channeling the conserved energy into the rapid pro- cells multiply in a confined area, however, the concentration of
duction of cell components such as DNA and protein. Famine, that molecule increases proportionally. Only when a signaling
178 Chapter 7 The Blueprint of Life, from DNA to Protein

Environmental stimulus

Sensor
protein

Bacterial cell Signaling

molecule

When few cells are present, the When many cells are present, the
concentration of the signaling signaling molecule reaches a
molecule is low. concentration high enough to induce
the expression of certain genes. Response
regulator
FIGURE 7.18 Quorum Sensing
The sensor protein spans the cytoplasmic membrane.
? Why would it be beneficial for cells to wait until a critical
The response regulator is a protein inside the cell.
population density is present before expressing certain genes?

molecule reaches a critical level does it induce the expression of

specific genes (figure 7.18).
Some types of bacteria are able to detect and even interfere with
the signaling molecules produced by other species. This allows them
to “eavesdrop” and even obstruct “conversations” of other bacteria.

Two-Component Regulatory Systems

An important mechanism that cells use to detect and react to changes
in the external environment is a two-component regulatory system P
(figure 7.19). E. coli uses such a system to control the expression
P
of genes for its alternative types of metabolism. When nitrate is
present in anaerobic conditions, the cells activate genes required
to use it as the terminal electron acceptor. Some pathogens use
In response to a specific change in the environment, the
two-component regulatory systems to sense and respond to envi- sensor phosphorylates a region on its internal portion.
ronmental magnesium concentrations. Because the magnesium The phosphate group is transferred to the response
concentration within certain host cells is generally lower than regulator, which can then turn genes on or off, depending
on the system.
that of the extracellular environment, these pathogens are able to
recognize whether or not they are within a host cell. In turn, they
can activate appropriate genes that help them evade host defenses. FIGURE 7.19 Two-Component Regulatory System
terminal electron acceptor, p. 130 ? E. coli cells use a two-component system to sense nitrate in the
Two-component regulatory systems consist of two different environment. What would happen if they lost the ability to sense
proteins—a sensor and a response regulator. The sensor spans the that compound?
cytoplasmic membrane. In response to specific environmental
variations, the sensor chemically modifies a region on its internal
portion, usually by phosphorylating a specific amino acid. The well-characterized examples is Neisseria gonorrhoeae. This bac-
phosphate group is then transferred to a response regulator. When terium has many different genes for pilin, the protein subunit that
phosphorylated, the response regulator can turn genes either on or makes up pili, yet most of these genes are silent. The only one
off, depending on the system. expressed is in a particular chromosomal location called an
expression locus. N. gonorrhoeae cells have a mechanism to
shuffle the pilin genes, randomly moving different ones in and out
of the expression locus. In a population of 104 cells, at least one is
Natural Selection expressing a different type of pilin. During an infection, the
Natural selection can also play a role in gene expression. The body’s immune system will begin to respond to the dominant pilin
expression of some genes changes randomly in cells, enhancing type, but the bacterial cells that have “switched” to produce a dif-
survival chances of at least a part of a population. ferent pilin type will survive and then multiply. Eventually,
The role of natural selection is readily apparent in bacteria the immune system learns to recognize those, but by that time,
that undergo antigenic variation, an alteration in the characteris- another subpopulation will have switched its pilin type.  pili, p. 65
tics of certain surface proteins. Pathogens that do this can stay one  Neisseria gonorrhoeae, p. 622
step ahead of the body’s defenses by altering the very molecules Another mechanism of randomly altering gene expression is
our immune systems must learn to recognize. One of the most phase variation, the routine switching on and off of certain genes.
 Part I Life and Death of Microorganisms 179

In E. coli, for example, certain types of pili required for attach- ■ Inducible. Inducible enzymes are not routinely produced at
ment to epithelial cells undergo phase variation. In an E. coli significant levels; instead, their synthesis can be turned on
population adhering to an epithelial surface, some bacterial cells when needed (figure 7.20). An example is β-galactosidase, the
will turn off the genes required for pili synthesis, thereby causing enzyme that hydrolyzes lactose into its component monosac-
those cells to detach from the surface. The process is reversible, so charides—glucose and galactose. The genes for this enzyme
the detached cells will later turn the genes on again, allowing the are part of the lac operon, which is turned on only when lactose
cells to colonize epithelial cells elsewhere. By altering the expres- is present. This makes sense because a cell would waste pre-
sion of genes such as these, at least a part of the population is cious resources if it expressed the operon when lactose is not
poised for change. available. Inducible enzymes are often involved in the transport
and breakdown of specific energy sources.
■ Repressible. Repressible enzymes are produced routinely,
MicroAssessment 7.5 but their synthesis can be turned off when they are not
Quorum sensing and two-component regulatory systems allow a cell required (figure 7.20). Repressible enzymes are generally
to respond to changing environmental conditions. The expression of involved in biosynthetic (anabolic) pathways, such as those
some genes changes randomly, increasing the chances of survival that produce amino acids. Cells require a sufficient amount
of at least a subset of a cell population under varying environmental of a given amino acid to multiply; so, if an amino acid is not
conditions. available in the environment, it needs to be produced by the
12. Explain how certain bacteria “sense” the density of cells. cell. When the amino acid is available, however, synthesis of
13. Describe antigenic variation. the enzymes used in its production would waste energy.
14. In quorum sensing, why might a bacterium synthesize more than
one type of signaling molecule? +
Mechanisms to Control Transcription
The methods a cell uses to prevent or facilitate transcription must
be readily reversible, allowing cells to control the relative number
7.6 ■ Bacterial Gene Regulation of transcripts made. Two of the most common regulatory mecha-
nisms are alternative sigma factors and DNA-binding proteins.
Learning Outcomes
9. Give an example of a constitutive enzyme, an inducible enzyme, Alternative Sigma Factors
and a repressible enzyme. As described earlier, sigma factor is a loose component of RNA
10. Using the lac operon as a model, explain the role of inducers, polymerase that functions in recognizing specific promoters.
repressors, and inducer exclusion. Standard sigma factors recognize promoters for genes that need to
be expressed during routine growth conditions, but a cell can also
In bacterial cells, many genes are routinely expressed, but others produce alternative sigma factors. These recognize different sets
are regulated in response to environmental conditions. Note that of promoters, thereby controlling the expression of specific groups
scientists describe these regulated genes as capable of being of genes. In the endospore-former Bacillus subtilis, the sporulation
turned on or off, but in reality there are no absolutes. In a popu- process is controlled by a number of different alternative sigma
lation of cells, a gene that is off may still be expressed at very
low levels.
A regulatory mechanism sometimes controls the transcrip-
tion of only a limited number of genes, but in other cases, a wide Inducible enzymes
These enzymes are not routinely
array of genes is controlled coordinately. A set of regulated produced, but mechanisms can
genes transcribed as a single polycistronic message is called an turn expression on for as long as
operon. One of the most well-characterized examples is the lac needed, for example, when the
Turn ON enzyme’s substrate is present.
operon, the set of genes required for transporting and hydro- (push and hold)
lyzing the disaccharide lactose. Separate operons controlled by a
single regulatory mechanism constitute a regulon. Two-
component regulatory systems often control regulons. The simul-
taneous regulation of numerous genes is called global control. Repressible enzymes
disaccharide, p. 31 These enzymes are routinely
When describing enzymes, scientists group them according to produced, but mechanisms can
turn expression off for as long as
the type of regulation that governs their synthesis: necessary, for example, when
the enzyme’s product is present
■ Constitutive. Constitutive enzymes are synthesized con- Turn OFF in sufficient quantity.
stantly; the genes that encode these enzymes are always (push and hold)
active. Constitutive enzymes usually play indispensable roles
in the central metabolic pathways. For example, the enzymes FIGURE 7.20 Principles of Regulation
of glycolysis are constitutive. central metabolic pathways, ? Why would the biosynthetic enzymes of a cell be repressible
pp. 132, 139 rather than constitutive or inducible?
180 Chapter 7 The Blueprint of Life, from DNA to Protein

factors. One controls the steps at the beginning of sporulation. With the repressor unable to bind to DNA, RNA polymerase
Others then guide the stages of development in the mother cell and may transcribe the gene.
spore. A cell can also express anti-sigma factors, which inhibit the ■ Repression. The repressor is synthesized as a form that
function of specific sigma factors. sporulation, p. 67 cannot bind to the operator. However, when a mole-
cule termed a corepressor attaches to the repressor, the
DNA-Binding Proteins corepressor-repressor complex can then bind to the operator,
Transcription is often controlled by proteins that bind to specific blocking transcription.
DNA sequences. When a regulatory protein attaches to DNA, it
Activators An activator is a regulatory protein that facilitates
can act either as a repressor, which blocks transcription, or an
transcription (positive regulation). Genes controlled by an activator
activator, which facilitates transcription.
have an ineffective promoter preceded by an activator-binding
Repressors A repressor is a regulatory protein that blocks tran- site. The binding of the activator to the DNA enhances the abil-
scription (negative regulation). It does this by binding to an opera- ity of RNA polymerase to initiate transcription at that promoter.
tor, a specific DNA sequence located immediately downstream of Like repressors, activators can be changed by the binding of other
a promoter. When a repressor is bound to an operator, RNA poly- molecules. When a molecule called an inducer binds to an activa-
merase cannot progress past that DNA sequence. Repressors are tor, the shape of the activator is altered so that it can now bind to
allosteric proteins, however, meaning that specific molecules can the activator-binding site (figure 7.22). Thus, the term “inducer”
attach to them and change their shape. This can alter the repressor’s applies to a molecule that turns on transcription, either by stimulat-
ability to bind to operator DNA. As shown in figure 7.21, there are ing the function of an activator or interfering with the function of
two general mechanisms by which different repressors can function: a repressor.
■ Induction. The repressor is synthesized as a form that binds
MicroByte
to the operator, blocking transcription. When a molecule In Bacillus subtilis, the DNA-binding protein that triggers the sporu-
called an inducer attaches to the repressor, the shape of the lation process controls a regulon of over 120 genes.
repressor changes so that it can no longer grasp the operator.

a Induction

Repressor +
Inducer helps turn
Transcription normally off. Inducer Repressor transcription on.

Operator
RNA polymerase
bound to promoter Transcription

Transcription blocked Inducer binds to the repressor and alters its shape, so that it can
The repressor binds to the operator, blocking transcription. no longer bind to the operator. The DNA is open for transcription.

b Repression

+
Repressor Corepressor helps
Transcription normally on. Corepressor Repressor turn transcription off.

Operator
RNA polymerase
Transcription
bound to promoter

The repressor alone cannot bind to the operator. The DNA is open Transcription blocked
for transcription. The corepressor-repressor complex can bind to the operator,
blocking transcription.

FIGURE 7.21 Transcriptional Regulation by Repressors (a) Induction. (b) Repression.

? How is a corepressor different from an inducer? How is it similar?
 Part I Life and Death of Microorganisms 181

+
RNA
polymerase Inducer Activator
Transcription (active) Inducer helps turn
Activator normally off. transcription on.
(inactive)

Activator-
binding site Promoter Transcription

RNA polym
polymerase cannot bind to the promoter unless the activator
ctivator Inducer
Ind binds
bin
nds to the
he activator and changes
c its shape, allowing
ng the
is bound to the activator-binding site, but the activator is in an activator to bind to the site. RNA polymerase can then bind to the
inactive form. promoter and initiate transcription.

FIGURE 7.22 Transcriptional Regulation by Activators

? How do activators facilitate transcription?

The lac Operon as a Model expression of the genes is significant biologically because it forces
cells to use the most efficiently metabolized carbon source first.
Originally described in the early 1960s by Francois Jacob and
Jacques Monod, the lac operon of E. coli has served as an impor-
tant model for understanding the control of bacterial gene expres- Lactose and the lac Operon
sion. This operon encodes proteins involved with the transport and The lac operon uses a repressor that prevents transcription when
degradation of lactose, and is only turned on when lactose is in the lactose is not available; the repressor binds the operator, blocking
cell but glucose is not available. The fact that glucose prevents RNA polymerase (figure 7.23). When lactose is in the cell,

No lactose in the cell

The repressor binds to operator, blocking transcription.

lacZ lacY lacA

(β-galactosidase) (permease) (transacetylase)

DNA Terminator
RNA polymerase Repressor bound
Repressor bou
b und
bound to promoter to ope
operator
perator
Lactose present in the cell
Some lactose is converted to allolactose. This binds to
the repressor and alters its shape, so that it can no
longer bind to the operator. If glucose is not available,
the operon will be transcribed.

Transcription

Transcription
Allolactose Non-functional
repressor

Translation

FIGURE 7.23 Lactose and the lac Operon

? What is the function of β-galactosidase? What is
the function of a permease?
182 Chapter 7 The Blueprint of Life, from DNA to Protein

however, some of it is converted to allolactose, an inducer. This One mechanism of carbon catabolite repression involves an
compound binds the repressor and, in doing so, changes the activator called CAP (catabolite activator protein), which is
repressor’s shape so that it can no longer grasp the operator. With required for transcription. To be functional, the activator must be
the operator unoccupied, RNA polymerase can begin transcribing bound by an inducer—an ATP derivative called cAMP (cyclic
the operon. However, this can happen only if glucose is not avail- AMP). The inducer is made only when extracellular glucose levels
able in the growth medium. are low, because the enzyme required for its synthesis is activated
by the idle form of the glucose transporter component.
Glucose and the lac Operon Although a great deal of attention has been paid to the role of
When glucose is available, the lac operon is not expressed because the activator in carbon catabolite repression, another mechanism
of a phenomenon called carbon catabolite repression (CCR). of regulation called inducer exclusion might be more significant
E. coli cells prioritize carbon/energy sources, a trait that can be in E. coli. In this mechanism, when glucose is being moved into
demonstrated by growing them in a medium containing both glu- the cell, a glucose transport component binds to the lactose trans-
cose and lactose. Initially, the cells multiply using only glucose. porter (permease), locking it in a non-functional position. The
Once the supply of that sugar is exhausted, growth stops for a locked permease cannot move lactose into the cell, so the lac
short period as the cells gear up to begin metabolizing lactose. operon will not be induced. Once the glucose supply diminishes,
Then, they begin multiplying again, this time using lactose to fuel the glucose transporter becomes idle, so lactose can then be
their growth. This characteristic two-phase growth pattern is brought into the cell.
called diauxic growth (figure 7.24).
Carbon catabolite repression is a global control system that MicroAssessment 7.6
allows glucose to regulate expression of the lac operon as well as Enzymes can be constitutive, inducible, or repressible. A repressor
other sets of genes. Glucose does not act directly in the regulation, blocks transcription when it binds to an operator. An activator
however. Instead, the cell’s glucose transport system serves as a enhances transcription when it binds to an activator-binding site. The
sensor of glucose availability. When the transport system is mov- lac operon, a model for regulation, is controlled by a repressor, an
ing glucose molecules into the cell, catabolite repression prevents activator, and inducer exclusion.
the lac operon from being expressed. When the transport system 15. Explain the difference between a constitutive enzyme and an
is idle, indicating that glucose is not available, then the lac operon inducible enzyme.
can be turned on (figure 7.25). 16. Explain how glucose represses the lactose operon.
17. Why would it be advantageous for a cell to control the activity of
an enzyme as well as its synthesis? +

Lactose
used up
7.7 ■ Eukaryotic Gene Regulation
Number of cells (logarithmic scale)

Learning Outcome
11. Describe how RNA interference silences genes.

Glucose Growth on
lactose
Considering the complexity of eukaryotic cells and the diversity
used up
of cell types found in multicellular organisms, it is not surpris-
ing that eukaryotic gene regulation is much more complicated
than that of prokaryotic organisms. Eukaryotic cells use a vari-
ety of control methods, including modifying the structure of the
Glucose and chromosome, regulating the initiation of transcription, and
lactose Growth on altering pre-mRNA processing and modification. We will focus
added glucose only on a process called RNA interference (RNAi), a recent
Nobel-winning discovery that revolutionized the current views
on gene regulation. Cells routinely use RNAi to destroy specific
RNA transcripts, and scientists can manipulate the process to
Time of incubation (hr)
silence select genes.
In RNAi, a cell produces short single-stranded RNA pieces to
FIGURE 7.24 Diauxic Growth Curve of E. coli Growing in a
Medium Containing Glucose and Lactose Cells preferentially locate specific RNA transcripts that need to be destroyed. To func-
use glucose. Only when the supply of glucose is used up do cells start tion in RNAi, a short RNA strand joins a multi-protein unit called
metabolizing lactose. Note that the growth on lactose is slower than it is an RNA-induced silencing complex (RISC). Within a RISC, the
on glucose. short RNA strand serves as the probe that allows the complex to
? Why would growth on lactose be slower than that on glucose? locate a specific nucleotide sequence on mRNA molecules. The
 Part I Life and Death of Microorganisms 183

Glucose transporter as a sensor Glucose transporter as a sensor

High glucose Low glucose

Glucose

Glucose
transporter

P R~ P ~P R~ P
P
R Unphosphorylated ~P R
P P transporter component

The unphosphorylated form of the The phosphorylated form of the

glucose transporter component indicates transporter component indicates that
that glucose is available in the medium. glucose is not available in the medium.
This is because the phosphorylated form This is because it cannot donate its
donates its phosphate group during the phosphate during glucose transport.
transport process.

Inducer exclusion Positive regulation of the lac operon

Posi
P
ATP
Glucose
Glucos
Glu
G cosee
Lactose R~ P
E. coli cellll
cAMP (inducer)
+

R
Lactose CAP Functional
transporter (inactive) activator
(permease)

High glucose
The unphosphorylated form of the
glucose transporter component
prevents the lactose transporter Low glucose
(permease) from functioning. Because The phosphorylated form of the glucose transporter component activates the
lactose cannot be moved into the cell, enzyme that produces cAMP, which binds to the activator (CAP). The complex of
the inducer (allolactose) cannot CAP and cAMP can then bind to the activator-binding site of the lac operon,
accumulate, so transcription will be permitting transcription. Note that lactose must be present for transcription to
blocked (see figure 7.23). occur (see figure 7.23).

FIGURE 7.25 Glucose and the lac Operon

? Why would it be advantageous for a cell to use glucose before lactose?

short RNA strand does this by binding to complementary MicroByte

sequences on an mRNA molecule, tagging that transcript for RNAi technology shows promise in treating certain viral infections
destruction by enzymes in the RISC (figure 7.26). The components and tumors.
of the RISC are not destroyed in the process, so the complex is
catalytic, providing a rapid and effective means of silencing genes MicroAssessment 7.7
that have already been transcribed. Two different types of RNA RNA interference uses short strands of RNA to locate specific RNA
molecules are used in RNAi—microRNA (miRNA) and short transcripts destined for destruction.
interfering RNA (siRNA). These are each about two dozen nucle-
18. What is the role of miRNA and siRNA in regulation of gene
otides in length and functionally equivalent, but they differ in how expression?
they are produced.
184 Chapter 7 The Blueprint of Life, from DNA to Protein

Analyzing a Prokaryotic DNA Sequence

Cell produces short single-stranded
RNA. When analyzing a DNA sequence, the (+) strand is used to repre-
sent the sequence of the corresponding RNA transcript. As an
example, an ATG in the (+) strand of DNA indicates a possible
start codon. plus (+) strand, p. 168
Computers help locate protein-encoding regions in DNA.
An RNA-induced silencing complex
(RISC) assembles. They search for open reading frames (ORFs), stretches of nucle-
otide sequences generally longer than 300 bp that begin with a
start codon and end with a stop codon. An ORF potentially encodes
RNA-induced silencing a protein. Other characteristics such as an upstream sequence that
complex (RISC) can serve as a ribosome-binding site also suggest that an ORF
encodes a protein.
The nucleotide sequence of an ORF can be compared with other
known sequences by searching computerized databases of published
sequences. The same can be done for the amino acid sequence of the
Binding of the RNA in the RISC to encoded protein. Not surprisingly, as genomes of more organisms
mRNA tags the mRNA for are being sequenced, information contained in these databases is
destruction. Enzymes cut mRNA; growing at a remarkable rate. If the encoded protein shows certain
RISC can then bind to another
mRNA molecule. amino acid similarities to characterized proteins, a presumed func-
tion can sometimes be assigned. For example, proteins that bind
DNA have similar amino acid sequences in certain regions.
Likewise, regulatory regions in DNA such as promoters can some-
FIGURE 7.26 RNA Interference (RNAi) times be identified based on similarities to known sequences.
? How could RNAi be used medically?

Metagenomics
Metagenomics is the analysis of total microbial genomes in an
7.8 ■ Genomics environment. By examining the total genomes, researchers can
study all microorganisms and viruses in a community, not just the
Learning Outcomes relatively few that grow in culture. Imagine the insights that can
12. Explain how protein-encoding regions are found when analyzing be gained from this new approach—tracking changes in the com-
a DNA sequence. position of the normal microbiota of a person over time to see if
13. Describe metagenomics and the information it can provide. there are changes during health and disease; comparing the micro-
biota of different body sites; and even comparing the microbiota
In 1995, the nucleotide sequence of the chromosome of of different people around the world! Metagenomics is also being
Haemophilus influenzae was published, marking the first com- used to study microbial life in the open oceans and in soils.
plete genomic sequence ever determined. Since then, sequencing Analyzing these sequences gives an entirely new perspective on
microbial genomes has become relatively common, leading to the extent of biodiversity and will probably lead to the discovery
many exciting advances, including a better understanding of the of new antibiotics and other medically useful compounds.
complex relationships between microbes and humans. In fact, Although metagenomics holds a great deal of promise, the
many of the recent findings described in this textbook have been amount of new data presents tremendous challenges. New com-
discovered through genomics. puting methods are being developed to handle the complications
Although sequencing methods are becoming more rapid, ana- of analyzing such complex information.
lyzing the resulting data is far more complex than it might initially
MicroByte
seem. Imagine trying to determine the amino acid sequence of a So far, over 1,000 prokaryotic genomes have been sequenced!
protein encoded by a stretch of DNA, without knowing anything
about the orientation of the gene’s promoter or the reading frame
used for translation. Because either strand of the DNA molecule MicroAssessment 7.8
could potentially be the template strand, two entirely different RNA
Sequencing methods are rapid, but analyzing the data and extracting
sequences must be considered as protein-encoding candidates. In the pertinent information is difficult.
turn, each of those two candidates has three reading frames, for a
19. What is an open reading frame?
total of six possible reading frames to consider. Yet only one of
these actually codes for the protein. Understandably, computers are 20. Describe two things that you can learn by searching a
computerized database for sequences that have similarities to a
an invaluable aid and are used extensively in deciphering the mean-
newly sequenced gene.
ing of the raw sequence data. As a result, a new field has emerged—
21. There are characteristic differences in the nucleotide sequences
bioinformatics—which creates the computer technology to store,
of the leading and lagging strands. Why might this be so? +
retrieve, and analyze nucleotide sequence data.
 Part I Life and Death of Microorganisms 185

FUTURE CHALLENGES 7.1

Gems in the Genomes?
From a medical standpoint, one of the most of those genes are understood, scientists should proteins of medical value. Previously unchar-
exciting challenges is to put the information be able to design drugs that turn off synthesis acterized proteins that have sequence similari-
obtained from sequencing microbial genomes of key proteins, making the organism harmless. ties to proteins of known therapeutic value are
into use. The potential gains are tremendous. Learning more about the human genome among the most promising. Some of these
For example, by studying the genomes of provides another means of developing drug are now in clinical trials to test their efficacy,
pathogenic microbes, scientists can learn more therapies. Already, companies are searching while other “gems” are probably still hidden,
about specific genes that enable an organism to genomic databases, a process called genome waiting to be discovered.
cause disease. Once the regulatory mechanisms mining, to locate ORFs that may encode

Summary
7.1 ■ Overview (figure 7.1) site serves as a start codon (figure 7.12). Ribosomes are translation
“machines.” tRNAs carry specific amino acids and act as keys that
Characteristics of DNA (figure 7.2)
interpret the genetic code (figure 7.14). The ribosome moves along
A single strand of DNA has a 5 end and a 3 end; the two strands of mRNA in the 5 to 3 direction; translation terminates when the ribo-
DNA in the double helix are complementary and antiparallel. some reaches a stop codon (figure 7.16).

Characteristics of RNA
7.4 ■ Differences Between Eukaryotic and Prokaryotic
A single-stranded RNA molecule is transcribed from one of the two
Gene Expression (table 7.5)
strands of DNA. There are three different functional types of RNA
molecules: messenger RNA (mRNA), ribosomal RNA (rRNA), and Eukaryotic mRNA is processed; a cap and a poly A tail are added.
transfer RNA (tRNA) (figure 7.3). Eukaryotic genes often contain introns; splicing removes these from
pre-mRNA (figure 7.17). In eukaryotic cells, the mRNA must be trans-
Regulating Gene Expression ported out of the nucleus before it can be translated in the cytoplasm.
Protein synthesis is generally controlled by regulating the synthesis
of mRNA (figure 7.4). 7.5 ■ Sensing and Responding to
Environmental Fluctuations
7.2 ■ DNA Replication
Signal Transduction
The bidirectional progression of replication around a circular DNA
Bacteria use quorum sensing to activate genes that are useful only when
molecule creates two replication forks (figure 7.5). DNA replication is
expressed by a critical mass (figure 7.18). Two-component regulatory
semiconservative.
systems use a sensor that recognizes changes outside the cell and then
transmits that information to a response regulator (figure 7.19).
Initiation of DNA Replication
DNA replication begins at the origin of replication. Natural Selection
The expression of some genes changes randomly, enhancing survival
The Process of DNA Replication
chances of at least a part of a population. Antigenic variation is a rou-
DNA polymerase synthesizes DNA in the 5 to 3 direction, using tine alteration in the characteristics of certain surface proteins. Phase
one strand as a template to generate the complementary strand variation is the routine switching on and off of certain genes.
(figures 7.6, 7.7).

7.3 ■ Gene Expression in Bacteria 7.6 ■ Bacterial Gene Regulation

Constitutive enzymes are constantly synthesized. The synthesis of
Transcription
inducible enzymes can be turned on by certain conditions. The syn-
RNA polymerase synthesizes RNA in the 5 to 3 direction, producing thesis of repressible enzymes can be turned off by certain conditions
a single-stranded RNA molecule complementary and antiparallel to the (figure 7.20).
DNA template (figure 7.9). Transcription initiates when RNA poly-
merase recognizes and binds to a promoter (figures 7.10, 7.11). When Mechanisms to Control Transcription
RNA polymerase encounters a terminator, it falls off the DNA and
Repressors block transcription (figure 7.21). Activators enhance tran-
releases the newly synthesized RNA.
scription (figure 7.22).
Translation The lac Operon as a Model
The information encoded by mRNA is deciphered using the genetic The lac operon uses a repressor that prevents transcription of the
code (table 7.4). The first AUG downstream of a ribosome-binding genes when lactose is not available (figure 7.23). A mechanism called
186 Chapter 7 The Blueprint of Life, from DNA to Protein

carbon catabolite repression (CCR) prevents transcription of the lac 7.8 ■ Genomics
operon when glucose is available (figure 7.25).
Analyzing a Prokaryotic DNA Sequence
When analyzing a DNA sequence, the (+) strand is used to represent the
7.7 ■ Eukaryotic Gene Regulation sequence corresponding RNA transcript; computers are used to search
for open reading frames (ORFs).
Regulation in eukaryotic cells is much more complicated than that in
prokaryotic cells. In RNA interference (RNAi), a cell synthesizes Metagenomics
short single-stranded RNA pieces to locate specific RNA transcripts Metagenomics allows researchers to study all microorganisms and
destined for destruction (figure 7.26). viruses in a community, not just the relatively few that grow in culture.

Review Questions
Short Answer 6. Which of the following statements about gene expression is false?
1. Explain what the term semiconservative means with respect to a) More than one RNA polymerase can be transcribing a specific
DNA replication. gene at a given time.
2. What is an origin of replication? b) More than one ribosome can be translating a specific transcript
at a given time.
3. Why are primers required in DNA replication but not in transcrip-
c) Translation begins at a site called a promoter.
tion?
d) Transcription stops at a site called a terminator.
4. What is polycistronic mRNA?
e) Some amino acids are coded for by more than one codon.
5. Explain why knowing the orientation of a promoter is critical when
determining the amino acid sequence of an encoded protein. 7. An enzyme used to synthesize the amino acid tryptophan is most
likely
6. What is the function of a sigma factor?
a) constitutive. b) inducible.
7. What is the fate of a protein that has a signal sequence?
c) repressible. d) a and b.
8. Explain how some bacteria sense the density of cells in their own
8. Under which of the following conditions will transcription of the
population.
lac operon occur?
9. Compare and contrast regulation by a repressor and an activator. a) Lactose present/glucose present
10. Explain why it is sometimes difficult to locate genomic regions b) Lactose present/glucose absent
that encode a protein. c) Lactose absent/glucose present
d) Lactose absent/glucose absent
Multiple Choice e) a and b
1. All of the following are involved in transcription except 9. All of the following are characteristics of eukaryotic gene
expression except
a) polymerase. b) primer. c) promoter.
a) 5 cap is added to the mRNA.
d) sigma factor. e) uracil.
b) a poly A tail is added to the 3 end of mRNA.
2. All of the following are involved in DNA replication except c) introns must be removed to create the mRNA that is translated.
a) polysome. b) gyrase. c) polymerase. d) the mRNA is often polycistronic.
d) primase. e) primer. e) translation begins at the first AUG.
3. All of the following are directly involved in translation except 10. Which of the following statements is false?
a) promoter. b) ribosome. c) start codon. a) A derivative of lactose serves as an inducer of the lac operon.
d) stop codon. e) tRNA. b) Signal transduction provides a mechanism for a cell to sense
the conditions of its external environment.
4. Using the DNA strand shown here as a template, what will be the
c) Quorum sensing allows bacterial cells to sense the density of
sequence of the RNA transcript?
like cells.
5 GCGTTAACGTAGGC 3
⎯→ d) An example of a two-component regulatory system is the
promoter
3 CGCAATTGCATCCG 5 lactose operon, which is controlled by a repressor and an
a) 5 GCGUUAACGUAGGC 3 activator.
b) 5 CGGAUGCAAUUGCG 3 e) An ORF is a stretch of DNA that may encode a protein.
c) 5 CGCAAUUGCAUCCG 3
d) 5 GCCUACGUUAACGC 3 Applications
1. A graduate student is trying to identify the gene coding for an enzyme
5. A ribosome binds to the following mRNA at the site indicated by
found in a bacterial species that degrades trinitrotoluene (TNT). The
the dark box. At which codon will translation likely begin?
student is frustrated to find that the organism does not produce the
5 ■ GCCGGAAUGCUGCUGGC enzyme when grown in nutrient broth, making it difficult to collect
a) GCC b) GGC the mRNA needed to help identify the gene. What could the student
c) AUG d) AAU do to potentially increase the amount of the desired enzyme?
 Part I Life and Death of Microorganisms 187

2. A student wants to remove the introns from a segment of DNA 2. In a variation of the experiment in the previous question, the fol-
coding for protein X. Devise a strategy to do this. lowing materials were added to three separate cell-free systems,
generating the indicated results:
Critical Thinking +
1. The study of protein synthesis often uses a cell-free system where
cells are ground with an abrasive to release the cell contents and Materials Added Results
then filtered to remove the abrasive. These materials are added to
Radioactive amino acids Radioactive protein produced
the system, generating the indicated results:
Radioactive amino acids Radioactive protein produced
Materials Added Results and DNase (a DNA-digesting
Radioactive amino acids Radioactive protein produced enzyme)
Radioactive amino acids No radioactive protein produced Several hours after grinding:
and RNase (an RNA-digesting Radioactive amino acids No radioactive protein produced
enzyme) and DNase

What is the best interpretation of these observations? What is the best interpretation of these observations?
8 Bacterial Genetics
KKE EYYT TE ERRMMS S
Auxotroph A microorganism that
requires an organic growth factor.
Conjugation Mechanism of
Non-Homologous
Recombination DNA
recombination that does not require
extensive nucleotide sequence
horizontal gene transfer in which the
similarity in the stretches that
donor cell physically contacts the
recombine.
recipient cell.
Phenotype The observed
DNA-Mediated Transformation
characteristics of a cell.
Mechanism of horizontal gene
transfer in which the bacterial DNA Plasmid An extrachromosomal
is transferred as “naked” DNA. DNA molecule that replicates
independently of the chromosome.
Genotype The sequence of
nucleotides in an organism’s DNA. Prototroph A microorganism that
does not require any organic growth
Homologous Recombination
factors.
Process by which a cell replaces a
stretch of DNA with a segment that Transduction Mechanism of
has a similar nucleotide sequence. horizontal gene transfer in which
bacterial DNA is transferred inside a
Horizontal Gene Transfer
phage coat.
DNA transfer from one bacterium
to another by conjugation, DNA- Transposon Segment of DNA that
mediated transformation, or can move from one site to another in
transduction. a cell’s genome.

A lysed E. coli cell shows the length of its unwound circular chromosome. Mutation A change in the Wild Type Form of the cell or gene
nucleotide sequence of a cell’s as it typically occurs in nature.
DNA that is then passed on to
daughter cells.
A Glimpse of History
Barbara McClintock (1902–1992) was a remarkable scientist who made
several very important discoveries in genetics. She carried out her studies
before the age of large interdisciplinary research teams and sophisticated
techniques of molecular genetics. Her tools consisted of a clear mind and
a curiosity that could make sense of confusing observations. She worked
12-hour days, 6 days a week in a small laboratory at Cold Spring Harbor
infections has been penicillin-like antibiotics, such as methicillin.
on Long Island, New York. Today, however, this treatment is likely to fail. In 2004, well
McClintock’s experimental system consisted of kernels in ears of over 60% of the S. aureus strains from hospitalized patients were
corn. She noticed that the various kernel colors were not inherited in a resistant to methicillin. In the United States, an estimated 2.3 mil-
predictable manner. In fact, the colors seemed to come and go. Based on lion healthy people harbor methicillin-resistant S. aureus (MRSA)
extensive data, McClintock concluded that segments of DNA, now called as part of their microbiota. Unfortunately, healthcare-associated
transposons, were moving into and out of genes involved with kernel color. MRSA strains (HA-MRSA) are also resistant to a variety of other
This destroyed the function of the genes, thereby changing kernel color. antibiotics. These are commonly treated with vancomycin, often
At the time that McClintock published her results in 1950, considered the drug of last resort. The situation became even more
most scientists believed that chromosomal DNA was very stable and worrisome in 2002, when S. aureus isolated from foot ulcers on a
changed only through recombination. Consequently, geneticists were
diabetes patient in Detroit was resistant to vancomycin in addition
skeptical of her conclusions. It was not until the late 1970s that her
earlier ideas began to be accepted. By that time, transposons had been
to other antibiotics.
discovered in many organisms, including bacteria. Although transpo- How do multidrug-resistant strains arise? How are these
sons were first discovered in plants, once they were found in bacteria, resistance traits transferred to other bacteria? The answer to
the field moved ahead very quickly. In 1983, at age 81, McClintock these and many other questions important to human health
received a Nobel Prize for her discovery of transposons, popularly requires a basic understanding of bacterial genetics. This sub-
called “jumping genes.” ject encompasses the study of heredity: how genes function
(covered in chapter  7), change, and are transferred to other

S
taphylococcus aureus, the Gram-positive coccus com- organisms. With this knowledge, you will understand why anti-
monly called Staph, is a frequent cause of skin and wound biotics are no longer the miracle drugs they once were against
infections. Since the 1970s, the usual treatment for these infectious diseases.
188
 Part I Life and Death of Microorganisms 189

8.1 ■ Genetic Change in Bacteria genetic makeup. For this reason, it is not surprising that we know
more about the genetics of the model organism E. coli than any
Learning Outcomes other organism in the world. model organisms, p. 8
1. Distinguish between genotype and phenotype.
A change in an organism’s DNA alters the genotype, the
sequence of nucleotides in the DNA. In bacterial cells, such a
2. Distinguish between mutation and horizontal gene transfer.
change can have a significant impact because bacteria are hap-
loid, meaning they contain only a single set of genes. No “backup
In the ever-changing conditions that characterize most environ- copy” of a gene exists in a haploid organism. Because of this, a
ments, all organisms need to adapt in order to survive and change in genotype often alters the organism’s observable char-
multiply. If they fail, competing organisms more “fit” to thrive in acteristics, or phenotype. Note, however, that the phenotype
the new setting will soon predominate. This is the process of involves more than just the genetic makeup of an organism; it can
natural selection. Bacteria have two general means by which they also be influenced by environmental conditions. For example,
routinely adjust to new circumstances: regulating gene expression colonies of Serratia marcescens are red when incubated at 22°C
(discussed in chapter 7) and genetic change, the focus of this but white when incubated at 37°C. The phenotype, but not the
chapter. bacterial gene regulation, p. 179 genotype, has changed. However, if the genes responsible for
Bacteria are an excellent experimental system for genetic pigment production are removed, the organism’s phenotype as
studies. They grow rapidly in small volumes of simple inexpen- well as the genotype changes.
sive media, accumulating in very large numbers. This makes it Genetic change in bacteria occurs by two mechanisms—
easy to study rare events that give rise to strains differing in their mutation and horizontal gene transfer (figure  8.1). Mutation

Spontaneous
mutation in genome

Vertical gene transfer

(a) Mutation

Plasmid transfer

Vertical gene transfer

(b) Horizontal gene transfer

FIGURE 8.1 Mechanisms of Genetic Change in Bacteria (a) Mutation. (b) Horizontal transfer of plasmid-encoded genes; other DNA can be
transferred horizontally as well.
? Which of these mechanisms would have the most pronounced effect on an organism’s genotype?
190 Chapter 8 Bacterial Genetics

changes the existing nucleotide sequence of a cell’s DNA, which MicroAssessment 8.1
is then passed on to the progeny (daughter cells) through vertical
gene transfer. The modified organism and daughter cells are The properties of bacteria can change either through mutation or
horizontal gene transfer.
referred to as mutants. Horizontal gene transfer is the acquisition
of genes from another organism. Like mutations, the changes are 1. How is mutation different from horizontal gene transfer?
then passed on to the progeny. 2. Contrast genotype and phenotype.
3. Which has a longer-lived effect on a cell—a change in the
genotype or a change in the phenotype? +

MUTATION AS A MECHANISM
OF GENETIC CHANGE
Mutations can change an organism’s phenotype. For example, if a necessarily identical. This gives a population the chance to adapt
gene required for biosynthesis of the amino acid tryptophan is to changing environments. The environment does not cause the
deleted, then the organism can multiply only if tryptophan is sup- mutations but selects those cells that can grow under its condi-
plied in the growth medium. The same occurs if the gene is dis- tions. For example, an organism with a spontaneous mutation
rupted so that the protein it encodes no longer functions properly. to antimicrobial resistance, though rare, will become dominant in
A mutant that requires a growth factor is an auxotroph (auxo an environment where the medication is present. This happens
means “increase,” and troph means “nourishment”). This is in because the antimicrobial kills the sensitive cells, allowing the
contrast to a prototroph, which does not require growth factors resistant cells to grow without competition.
(proto means “earliest form of”). growth factor, p. 93 A single mutation is a rare event, so two mutations are even
Geneticists working with mutants compare them to wild more unlikely. Physicians take advantage of this to prevent patho-
type, the typical phenotype of strains isolated from nature. A gens from developing resistance to certain antimicrobial medi-
wild-type E. coli strain is a prototroph. By convention, a strain’s cations. In tuberculosis treatment, for example, two or more
characteristics are designated by three-letter abbreviations, with antimicrobial drugs are given simultaneously. Any mutant bacte-
the first letter capitalized. For example, a strain that cannot make rium resistant to one medication is probably still sensitive to the
tryptophan is designated Trp–. For simplicity, only required other. The chance of a single cell becoming resistant spontane-
growth factors are indicated. Likewise, only if a cell is resistant ously to both medications is the product of the mutation rates of
to an antibiotic is it indicated; streptomycin resistance is indi- the two genes (calculated by taking the sum of the exponents). For
cated as StrR. example, if the mutation rate to “antibiotic X” resistance is 10–6
per cell division and the mutation rate to “antibiotic Y” resistance
is 10–8, then the probability that both mutations will spontaneously
8.2 ■ Spontaneous Mutations happen within the same cell is 10–6 × 10–8, or 10–14.

Learning Outcomes Base Substitution

3. Describe three outcomes of base substitutions. Base substitution, the most common type of mutation, occurs
4. Describe the consequences of removing or adding nucleotides. during DNA synthesis when an incorrect nucleotide is incorpo-
5. Explain how transposons cause mutations. rated (figure 8.2). If only one base pair is changed, the mutation
is called a point mutation.
Spontaneous mutations are genetic changes that result from nor- Base substitution leads to three possible mutation outcomes:
mal cell processes. They occur randomly, and genes mutate spon- silent, missense, or nonsense (figure 8.3). A silent mutation has a
taneously at infrequent but characteristic rates. The mutation rate codon that still specifies the wild-type amino acid. A missense
is defined as the probability that a mutation will occur in a given mutation results when the altered codon specifies a different amino
gene per cell division. The mutation rate of different genes usually acid. The effect of this depends on the position and the nature of
varies between 10–4 and 10–12 per cell division. In other words, the the change. In many cases, cells with a missense mutation grow
chance that a gene will undergo a mutation when a cell replicates slowly because the encoded protein functions only partially. Such a
its DNA prior to cell division is between one in 10,000 (10–4) and mutation is termed leaky. A nonsense mutation occurs when the
one in a trillion (10–12). exponents, Appendix 1, p. A-1 altered codon is a stop codon, resulting in a shorter and often non-
Mutations are passed on to a cell’s progeny. On rare occa- functional protein. Any mutation that totally inactivates the gene is
sions, however, a mutation will then change back to its original, termed a null or knockout mutation. Note that geneticists sometimes
non-mutant state. This change is termed reversion and, like the use the term “silent mutation” to indicate a mutation that does not
original mutation, occurs spontaneously at low frequencies. alter the function of the protein. With this broad definition, any base
Because spontaneous mutations occur routinely, every large substitution that does not affect the phenotype would be a silent
population contains mutants, so the cells in a colony are not mutation. codon, p. 170 stop codon, p. 173
 Part I Life and Death of Microorganisms 191

Base substitutions are more common in aerobic than anaero-

5' 3'
bic environments. This is because reactive oxygen species (ROS) • • • • • • T G T • • • • • • DNA
such as superoxide and hydrogen peroxide are produced from O2. • • • • • • A C A • • • • • •
3' 5'
These chemicals can oxidize the nucleobase guanine, and DNA
polymerase often mispairs oxidized guanine with adenine rather
than cytosine. reactive oxygen species, p. 90 nucleobase, p. 32
T G T T G C T G G T G A Mutation
A C A A C G A C C A C T
Deletion or Addition of Nucleotides
Deletion or addition of nucleotides during DNA replication also
results in spontaneous mutations. The consequence of this depends
on how many nucleotides are involved. If three nucleotide pairs Transcribed codon
UG U UG C UG G UG A
are added (or deleted), this adds (or deletes) one codon. When the
gene is expressed, one additional (or fewer) amino acid will be in
the resulting protein. How serious the effect of this change is Cysteine Cysteine Tryptophan Stop codon Amino acid translated
depends on its location in the encoded protein.
Wild type Silent Missense Nonsense Outcome
Adding or subtracting one or two nucleotide pairs causes a mutation mutation mutation
frameshift mutation (figure 8.4). This changes the reading frame of
the corresponding mRNA molecule so that an entirely different set
FIGURE 8.3 Potential Outcomes of Base Substitutions
of codons is translated. Frequently, one of the resulting downstream
codons will be a stop codon. As a consequence, a frameshift muta- ? Which of these outcomes is most likely to result in a leaky
mutation?
tion likely results in a shortened non-functional protein—a knockout
mutation. reading frame, p. 171 downstream, p. 169

Wild type
5' 3' 5' 3'
A AG C G G T A C G T T A A A
T T C DNA
G C C A T G C A A T T T
3' 5' 3' 5'
DNA strand
separation
5' 3'
5' 3' CG G U A C GU U A A A Transcribed codons
DNA A AG
replication
T T C
3' 5' DNA replication, an incorrect Arginine Tyrosine Valine Lysine Amino acids translated
nucleotide is incorporated

5' 3' 5' 3'

A AG A GG
Mutant
T T C T T C Base substitution
3' 5' 3' 5' Base pair
addition
Wild type DNA strand
separation 5' 3'
C G G A T A C G T T A A A DNA
5' 3' G C C T A T G C A A T T T
DNA A GG 3' 5'
replication,
generating T T C
a mutation 3' 5' 5' 3'
DNA replication CG G A U A CG U U A A Transcribed codons
5' 3' 5' 3'
A G G A AG
T C C T T C Arginine Isoleucine Arginine STOP Amino acids translated
3' 5' 3' 5'
Mutant Wild type
FIGURE 8.4 Frameshift Mutation as a Result of Nucleotide
Addition The addition of a nucleotide pair (base pair) to the
FIGURE 8.2 Base Substitution A replication error results DNA results in a shift in the reading frame when the sequence is
in a mismatch between the two DNA strands. Subsequent DNA transcribed and translated. Deletion of a single nucleotide pair in the
replication using the altered strand as template results in a point DNA would have a similar effect.
mutation.
? This figure shows a single nucleotide addition. What would
? What enzyme incorporated the incorrect nucleotide? happen if three nucleotides were added?
192 Chapter 8 Bacterial Genetics

Transposition
Transposable
element
Gene X Gene X
disrupted

FIGURE 8.5 Transposition A transposable element has the ability to “jump” (transpose) from one piece of DNA into another.
? What effect does the transposon have on the function of gene X?

Transposons (Jumping Genes) MicroAssessment 8.2

Transposons, or jumping genes, are segments of DNA that Spontaneous mutations happen during normal cell processes and can
can move from one location to another in a cell’s genome, alter the properties of the cell. A leaky mutation results in a partially
a process called transposition (figure 8.5). The gene into functional protein; a knockout mutation results in a non-functional
which a transposon jumps is insertionally inactivated by the protein. Base substitutions that occur during DNA synthesis can lead
to silent, missense, and nonsense mutations. Removing or adding
event, meaning that the insertion destroys the function of the
nucleotides can cause a frameshift mutation. Transposons can “jump”
gene. Most transposons contain transcriptional terminators, so from one location to another.
the expression of downstream genes in the same operon will
4. Which is generally more severe—a missense mutation or a
stop as well. The structure and biology of transposons will be
nonsense mutation?
described later in this chapter. transcriptional terminators, p. 170
5. What is the likely consequence of a frameshift mutation?
operon, p. 179
The classic studies of transposition were carried out 6. Is it as effective to take two antibiotics sequentially as it is
to take them simultaneously, as long as the total length of time
by Dr. Barbara McClintock (see A Glimpse of History). She
that they are both taken is the same? Explain. +
observed color variation in corn kernels as a result of transpo-
sons moving into and out of genes that control pigment syn-
thesis (figure 8.6).
8.3 ■ Induced Mutations
Learning Outcomes
6. Describe the three general groups of chemical mutagens.
7. Explain why transposons induce mutations.
8. Explain how X rays and UV light damage DNA.

Induced mutations are genetic changes that occur due to an influ-

ence outside of a cell, such as exposure to a chemical or radiation.
An agent that induces the change is a mutagen (table  8.1).
Geneticists—who depend on mutants to study cellular processes—
often use mutagens to increase the mutation rate in bacteria, mak-
ing mutants easier to find.

Chemical Mutagens
Some chemical mutagens cause base substitutions, and others
cause frameshift mutations.

Chemicals That Modify Nucleobases

A number of different chemicals modify the nucleobases in DNA,
FIGURE 8.6 Transposition Detected by Changes in Seed changing their base-pairing properties. This increases the chance
Color Variegation in the color of corn kernels is caused by insertion that the incorrect nucleotide will be incorporated during DNA
of transposable elements into genes involved in pigment synthesis. replication. For example, nitrous acid (HNO2) converts cytosine to
 Part I Life and Death of Microorganisms 193

TABLE 8.1 Common Mutagens

Agent Action Result

Chemical Agent
Chemicals that modify nucleobases
Nitrous acid Converts cytosine to uracil Nucleotide substitution
Alkylating agents Adds alkyl groups (CH3 and others) to nucleobases Nucleotide substitution
Base analogs Used in place of normal nucleobases in DNA Nucleotide substitution
5-Bromouracil
Intercalating agents Inserts between base pairs Addition or subtraction of nucleotides
Ethidium bromide
Transposons Randomly insert into DNA Insertional inactivation
Radiation
Ultraviolet (UV) light Causes intrastrand thymine dimer to form Errors during repair process
X rays Cause single- and double-strand breaks in DNA Deletions

uracil, which base-pairs with adenine rather than guanine. It Base Analogs
removes amino groups from adenine and guanine as well. A group Base analogs structurally resemble nucleobases but have different
of chemicals called alkylating agents adds alkyl groups (short hydrogen-bonding properties. The analogs can be mistakenly used
chains of carbon atoms) onto nucleobases. An example is nitroso- in place of the nucleobases when nucleotides are made, and DNA
guanidine, which adds a methyl group to guanine, causing it to polymerase then incorporates these into DNA. When a DNA
base-pair with thymine (figure 8.7). strand has a base analog, the wrong nucleotide can be incorporated
when the complementary strand is synthesized. For example,
MicroByte
Many chemical mutagens are used in cancer therapy to kill rapidly dividing 5-bromouracil resembles thymine, but it often base-pairs with
cancer cells. Unfortunately, they also damage DNA in normal cells. cytosine; 2-amino purine resembles adenine but often pairs with C
instead of T (figure 8.8). DNA replication, p. 165

FIGURE 8.7 Mutagenic Effects of the Alkylating Agent

Hydrogen bonds formed Nitrosoguanidine (a) Nitrosoguanidine converts guanine bases
with complementary bases Added
CH3 in DNA to methylguanine, which can base-pair with thymine
alkyl group
(T) as well as cytosine (C). (b) The altered base-pairing property
O O of methylguanine can result in point mutations following DNA
N H N replication.
N N
H H H H ? What are the three possible outcomes of point mutations?

N N N N N N
Alkylating agent
H H
deoxyribose deoxyribose

Guanine Methylguanine
(pairs with C) (sometimes pairs with T)

(a)

Wild type Nitroso- Mutant

5' 3' guanidine 5' 3' DNA 5' 3' DNA 5' 3'
A G T treatment A G* T replication A G* T replication A A T
T C A T C A T T A T T A
3' 5' 3' 5' 3' 5' 3' 5'

Guanine (G) is converted Thymine (formerly paired

Methylguanine of template
to methylguanine (G*) with G*) now serves as
strand pairs with thymine (T)
template and pairs with
instead of cytosine (C)
adenine (A)

(b)
194 Chapter 8 Bacterial Genetics

Thymine Thymine dimer

Normal
Analog
nucleobase
Thymine Covalent
O O bonds
H3C 6 H Br 6 H
5 1N Replaced by 5 1N
4 2 4 2
3 3
N O N O
H H Ultraviolet
Thymine 5-bromouracil light
(pairs with A) (pairs with A or C) Sugar-phosphate
backbone
H H
N N
7 6 N FIGURE 8.9 Thymine Dimer Formation UV light causes covalent
N 5 1
7 6 N 8 bonds to form between adjacent thymine molecules on the same
5 4 2 H
8
1 Replaced by 9 3 strand of DNA, distorting the shape of the DNA.
4 2
N N N
9 3 H
N N H ? What effect does a thymine dimer have on DNA synthesis?
H
H 2-amino purine
Adenine (pairs with T or C) Ultraviolet Light
(pairs with T)
Irradiation of cells with ultraviolet light causes covalent bonds to
form between adjacent thymine molecules on a DNA strand, pro-
FIGURE 8.8 Two Base Analogs Used in Mutagenesis The ducing thymine dimers (figure 8.9). The dimer cannot fit properly
altered base-pairing properties of the analogs can lead to point
mutations by the same mechanism shown in figure 8.7.
into the double helix, distorting the DNA molecule. Replication
and transcription stall at the distortion, and as a result, the cells will
? What base-pair substitution would 5-bromouracil generate?
die if the damage is not repaired. How then can UV light be muta-
genic? UV mutagenizes cells indirectly. Its major mutagenic action
results from the cell’s attempting to repair the damage by SOS
Intercalating Agents repair, described in the next section.
Intercalating agents increase the frequency of frameshift muta-
tions. They do this because they are flat molecules that can inter- X Rays
calate (insert) between adjacent base pairs in a strand of DNA. X rays cause single- and double-strand breaks in DNA, and altera-
This pushes the nucleotides apart, producing enough space tions to the nucleobases. Double-strand breaks often result in dele-
between bases that errors are made during replication. If the inter- tions that are lethal to the cell.
calating agent inserts into the template strand, a base pair will be
added as the new strand is synthesized. If it intercalates into the
MicroAssessment 8.3
strand being synthesized, a base pair will be deleted. As in spon-
taneous frameshift mutants, adding or subtracting a nucleotide in Mutagens increase the frequency of mutations. Some chemical
mutagens cause base substitutions, but intercalating agents cause
DNA often results in a stop codon being generated prematurely
frameshift mutations. Mutations from UV light are due to SOS repair;
in the mRNA transcript, giving rise to a shortened protein. X rays cause breaks in DNA strands and alter nucleobases.
Ethidium bromide, a chemical commonly used to stain DNA
7. How does an intercalating agent cause mutations?
in the laboratory, is an intercalating agent. The manufacturer
now warns users that the chemical should be handled with great 8. What mutagen causes thymine dimers, and why does it kill cells?
care because it likely is a carcinogen (cancer-causing). Another 9. Do you think mutations caused by reactive oxygen species should
intercalating agent is chloroquine, which has been used for many be considered spontaneous or induced? Justify your answer. +
years to treat malaria.

8.4 ■ Repair of Damaged DNA

Transposition
Transposons can be introduced intentionally into a cell in order to Learning Outcomes
generate mutations. The transposon, which cannot replicate on its 9. Explain how errors in nucleotide incorporation can be repaired
own because it lacks an origin of replication, inserts into the cell’s and the role of methylation in the process.
genome. This generally inactivates the gene into which it inserts 10. Explain how modified nucleobases can be repaired.
(see figure 8.5). 11. Describe three mechanisms cells use to repair thymine dimers.

The amount of spontaneous and mutagen-induced damage to

Radiation DNA is enormous. This damage, if not repaired, can quickly lead
Two kinds of radiation are commonly used as mutagens: ultravio- to cell death and, in animals, cancer. In humans, two breast cancer
let (UV) light and X rays. wavelengths of radiation, p. 115 susceptibility genes code for enzymes that repair damaged DNA.
 Part I Life and Death of Microorganisms 195

Mutations in either gene result in a high probability (80%) of Proofreading by DNA Polymerase
breast cancer. DNA polymerases are complex enzymes that not only synthesize
Mutations are rare because alterations in DNA are generally DNA, but also verify the accuracy of their actions—a character-
repaired before they can be passed on to progeny. Over the many istic called proofreading. The enzymes can back up and excise
millions of years of evolution, cells have developed several dif- (remove) a nucleotide not correctly hydrogen bonded to the
ferent DNA repair mechanisms (table 8.2). Eukaryotic and pro- opposing nucleobase in the template strand. The DNA poly-
karyotic cells share many of the same general mechanisms. Note merase then incorporates the correct nucleotide. Although the
that cells cannot repair all types of mutations, such as insertional proofreading function of DNA polymerases is very efficient, it is
inactivation caused by transposition. not flawless.
MicroByte
Every 24 hours, the genome of every cell in the human body is
damaged more than 10,000 times. Mismatch Repair
Mismatch repair fixes errors missed by the proofreading of DNA
polymerase. A specific protein binds to the site of the mismatched
Repair of Errors in Nucleotide nucleobase, directing an enzyme to cut the sugar-phosphate
Incorporation backbone of the strand. Another enzyme then degrades a short
DNA polymerase sometimes incorporates the wrong nucleotide region of that DNA strand, thereby eliminating the misincorporated
as it replicates DNA. The resulting mispairing of nucleobases nucleotide. How does the cell know which strand to remove? This
results in a slight distortion in the DNA helix, which can be is an important question because if the enzyme cuts the template
recognized by enzymes within the cell that then repair the mis- strand and not the new one, then the misincorporated nucleotide
take. By quickly repairing the error before the DNA is repli- would remain. The key to the answer lies in methylation of the
cated, the cell prevents the mutation. Two mechanisms for this DNA nucleobases. Soon after a DNA strand is synthesized, an
are proofreading by DNA polymerase and mismatch repair. enzyme adds methyl groups to certain nucleobases. This takes
 DNA polymerase, p.166 time, however, so the new strand is still unmethylated immediately

TABLE 8.2 Repair of Damaged DNA

Type of Defect Repair Mechanism Biochemical Mechanism Result

Spontaneous Wrong nucleotide Proofreading by DNA Mispaired nucleotide is Potential mutation

incorporated during DNA polymerase removed by DNA polymerase. eliminated
replication
Mismatch repair A protein binds to the site Potential mutation
of mismatch and cuts the eliminated
unmethylated strand. A short
stretch of that strand is then
degraded and DNA polymerase
synthesizes a replacement.
Oxidized guanine in DNA Action of glycosylase Glycosylase removes the Potential mutation
oxidized guanine. A short eliminated
stretch of that strand is then
degraded, and DNA polymerase
synthesizes a replacement.
Mutagen-Induced
Chemical Wrong nucleotide Proofreading and Same as for spontaneous Potential mutation
incorporated during DNA mismatch repair mutations (see proofreading eliminated
replication and mismatch repair)
UV light Thymine dimer formation Photoreactivation Breaks the covalent bond Original DNA
(light repair) between the thymine molecule restored
molecules
Excision repair A short stretch of the strand Potential mutation
(dark repair) containing the thymine dimer eliminated
is removed; DNA polymerase
then synthesizes a replacement.
SOS repair A special DNA polymerase Cell survives but
synthesizes DNA even when the numerous mutations
template is damaged. produced
196 Chapter 8 Bacterial Genetics

1 Template strand 1 5' 3' DNA contains oxidized

C T A A G–O C T guanine (G–O) as a result
CH3 CH3 G A T T C G A
5' 3' The wrong nucleotide 3' 5' of oxidation damage.
C T A A G C T G A G is incorporated during
G A T T T G A C T C
3' 5' DNA synthesis.
2 5' 3' Glycosylase removes the
Newly synthesized strand C T A A C T
G A T T C G A oxidized nucleobase from the
3' 5' sugar-phosphate backbone.
2 CH3 CH3
5' 3' Near the site of the mismatched
C T A A G C T G A G base, an enzyme cuts the 3 Cut
G A T T T G A C T C sugar-phosphate backbone 5' 3' At the site of the missing
3' 5' of the unmethylated strand. C T A A C T nucleobase, an enzyme cuts the
G A T T C G A
Cut 3' 5' sugar-phosphate backbone.

3 CH3 CH3 5' 3'

4
5' 3' C T C T DNA polymerase degrades a
An enzyme degrades a
C T A A G C T G A G G A T T C G A short stretch of the strand.
G A C T C short stretch of the strand
that had the error. 3' 5'
3' 5'

5 5' 3'
4 CH3 CH3 The combined actions of DNA
C T A A G C T
5' 3' G A T T C G A polymerase and DNA ligase
DNA polymerase synthesizes
C T A A G C T G A G 3' 5' fill in and seal the gap.
G A T T C G A C T C a new stretch, incorporating
3' 5' the correct nucleotide.
FIGURE 8.11 Repair of Oxidized Guanine
? What would be the effect on cells if they did not have the
5 CH3 CH3 glycosylase enzyme?
5' 3' DNA ligase joins the 3' end
C T A A G C T G A G of the newly synthesized
G A T T C G A C T C segment to the original
3' 5' strand. MicroByte
DNA ligase
In human cells, defects in either mismatch repair or repair of
modified nucleobases increases the incidence of colon cancers.
FIGURE 8.10 Mismatch Repair
? What role does methylation play in mismatch repair? Repair of Thymine Dimers
Bacteria have several mechanisms to combat the DNA-damaging
effects of UV light, a component of sunlight. In one mechanism, an
enzyme uses the energy of visible light to break the covalent bonds
after it is synthesized. Therefore, the template strand is methyl- of the thymine dimer, restoring the DNA to its original state
ated, whereas the new strand is not, allowing the repair enzyme to (figure  8.12). Because light is required for this mechanism, it is
distinguish between the two (figure  8.10). After the nucleotides called photoreactivation, or light repair. The enzyme used is found
are removed from the new strand, the combined actions of DNA only in prokaryotes.
polymerase and DNA ligase then fill in that section and seal the Some bacteria have an enzyme that recognizes the major distor-
gap. DNA ligase, p. 167 tions in DNA that result from thymine dimer formation. In this pro-
cess, excision repair, or dark repair, the enzyme makes single-stranded
Repair of Modified cuts that flank both sides of the damaged region to remove the strand
Nucleobases in DNA (figure 8.12). The actions of DNA polymerase and DNA ligase then
fill in and seal the gap left by the removal of the segment.
Modified nucleobases such as oxidized guanine can result in base
substitutions if they are not repaired before the DNA is replicated.
An important mechanism for repairing this defect uses a glycosyl- SOS Repair
ase, an enzyme that removes the oxidized nucleobase from the SOS repair is a last-ditch repair mechanism that copes with
sugar-phosphate backbone (figure  8.11). Another enzyme then extensively damaged DNA. The enzymes that carry out this repair
recognizes that a nucleobase is missing and cuts the DNA at this are induced when DNA is so heavily damaged by UV light that
site. DNA polymerase degrades a short section of this strand to photoreactivation and excision repair may not be able to correct all
remove the damage. This same enzyme synthesizes another strand of the damage. DNA and RNA polymerases then stall at sites of
with the proper nucleotides, and DNA ligase seals the gap in the unrepaired damage, so the cells cannot replicate or transcribe their
single-stranded DNA. DNA. Without SOS repair, the cells would die.
 Part I Life and Death of Microorganisms 197

Damaged DNA activates expression of the several dozen 8.5 ■ Mutant Selection
genes that encode the SOS system. One component of this system
is a DNA polymerase that synthesizes DNA even in extensively Learning Outcomes
damaged regions. Unlike the standard DNA polymerases, how- 12. Compare and contrast direct and indirect selection.
ever, the SOS DNA polymerase has no proofreading ability.
13. Describe how direct selection is used to screen for possible
Errors are made as a result, a process called SOS mutagenesis. carcinogens.

MicroAssessment 8.4 Mutations are rare events, even when mutagens are used,
DNA polymerases have proofreading ability. Mismatch repair fixes errors which presents a challenge to a scientist who wants to isolate
missed by the proofreading mechanism; methylation distinguishes the a desired mutant. In a culture containing several billion cells,
template strand. Specific glycosylases can remove modified nucleobases. perhaps only one cell has the sought-after mutation. How can
Thymine dimers can be repaired through photoreactivation and excision
that rare cell be isolated and identified? Depending on the type
repair; severe damage can be overcome by the SOS repair system.
of mutant being sought, either direct or indirect selection can
10. Distinguish between photoreactivation and excision repair of
be used.
thymine dimers.
11. How does UV light cause mutations?
12. To maximize the number of mutations following UV Direct Selection
irradiation, should you incubate the irradiated cells in the Mutants that can grow under conditions in which the parent
light or in the dark, or does it make any difference? Explain
your answer. +
cells cannot are usually easy to isolate by direct selection. In
this method, cells are inoculated onto an agar medium that sup-
ports the growth of the mutant, but not the parent. For example,
Photoreactivation antibiotic-resistant mutants can be easily selected directly by
inoculating cells onto a medium containing the antibiotic. Only
Covalent bonds the resistant cells will form colonies (figure 8.13).
5' 3' Thymine dimer distorts the
G C G A TT G A C G
C G C T AA C T G C DNA molecule. Indirect Selection
3' 5'
Indirect selection is used to isolate an auxotroph from a proto-
trophic parent strain. This process is more difficult than direct
5' 3' An enzyme uses visible light to selection because no medium will allow the growth of the
G C G A T T G A C G break the covalent bond of the mutant and not the parent. For example, Trp– mutants can grow
C G C T A A C T G C thymine dimer, restoring the DNA
3' 5' to its original state.

Excision repair
Covalent bonds
5' 3' Thymine dimer distorts the
G C G A TT G A C G
C G C T AA C T G C DNA molecule.
3' 5' Streptomycin- Streptomycin-
resistant mutant sensitive cells
Streptomycin-
resistant mutant
Cut Cut
Streptomycin- (looks the same
A T T GA resistant mutant as other colonies)
G An enzyme removes the damaged
5' 3'
G C C G section by cutting the DNA
C G C T A A C T G C backbone on either side of the
3' 5' thymine dimer.

5' 3' The combined actions of DNA

G C G A T T G A C G polymerase and DNA ligase fill in
C G C T A A C T G C Medium containing Medium without
3' 5' and seal the gap. streptomycin streptomycin

FIGURE 8.12 Repair of Thymine Dimers In photoreactivation, FIGURE 8.13 Direct Selection of Mutants Only cells carrying
an enzyme uses the energy of light to break the covalent bonds. In a mutation that confers resistance to streptomycin can grow on the
excision repair, the section containing the dimer is replaced. selective medium used here.
? How is the mechanism of excision repair similar to that of ? Direct selection cannot be used to isolate auxotrophic mutants in
mismatch repair? a culture of prototrophic parent cells. Why not?
 198 Chapter 8 Bacterial Genetics

only on a complex medium such as nutrient agar because this is first spread onto a nutrient agar plate. Mutant and non-mutant
supplies the tryptophan they require, but Trp+ parent cells also cells will grow on this medium to form colonies, creating what is
grow on this same medium. To overcome this problem, a tech- referred to as the master plate. 1 The master plate is then pressed
nique called replica plating is used, sometimes preceded by onto sterile velvet, a fabric with tiny threads that stand on end
penicillin enrichment of mutants. complex medium, p. 94 nutrient like small bristles. The velvet picks up some cells of every colony.
agar, p. 94 2 Next, two agar plates—one nutrient agar and one glucose-salts
agar—are pressed in succession and in the same orientation onto
the same velvet. This transfers cells taken from the master plate to
Replica Plating both the nutrient agar and the glucose-salts agar, creating replica
Replica plating is a clever method for indirect selection of auxo- plates. A line on the plates is used to maintain a consistent orienta-
trophic mutants, devised by Joshua and Esther Lederberg in the tion. 3 The replica plates are then incubated, allowing cells to
early 1950s (figure 8.14). In this technique, the bacterial culture grow to form colonies; prototrophs grow on both types of media,

1 A plate of bacterial colonies is pressed onto the 2 Cells adhering to the velvet are transferred to the sterile media, resulting in exact replicas
surface of sterile velvet. of the original plate.

Master plate with bacterial colonies Sterile plate; nutrient agar Sterile plate; glucose-salts agar
(nutrient agar)

Pressed onto Pressed to velvet

sterile velvet
Sterile velvet

Colonies imprinted Plates incubated Position of

on velvet Auxotroph missing
auxotroph

Nutrient agar; Glucose-salts agar;

all colonies grow. auxotrophs do not grow.

3 Auxotrophic mutants form colonies on the nutrient agar but not on the glucose-salts agar.

FIGURE 8.14 Indirect Selection of Mutants by Replica Plating The procedure shown was used by the Lederbergs and continues to be
used today in many laboratories.
? Why go to the trouble of creating a master plate (why not simply plate the initial culture on both nutrient agar and glucose-salts agar)?
 Part I Life and Death of Microorganisms 199

but auxotrophs grow only on the nutrient agar; recall that glucose- penicillinase is then added to destroy the penicillin, and the
salts is a chemically defined medium with no added growth factors cells plated on nutrient agar to create the master plate used in
(see table 4.7). glucose-salts, p. 95 replica plating. penicillin, p. 463
The plates are exact replicas, so colonies on the master
plate that cannot grow on glucose-salts can be identified; these
are auxotrophs. The particular growth factor required can then Screening for Possible Carcinogens
be determined by adding nutrients individually to a glucose- A substantial proportion of cancers appear to be caused by
salts medium and determining which one promotes growth. chemicals that are carcinogens (meaning “cancer generating”),
and most carcinogens are mutagens. Thousands of chemicals—
including pesticides, herbicides, hair dyes, cosmetics, food addi-
Penicillin Enrichment of Mutants tives, and the by-products of manufacturing processes—are
Penicillin enrichment is sometimes used before replica plat- released into the environment, so how can they all be tested for
ing to increase the proportion of auxotrophs in a broth culture. carcinogenic activity? Testing in animals for tumor formation
This is helpful because even when mutagenic agents are used, takes 2 to 3 years and may cost $100,000 or more to test a single
the frequency of mutations in a particular gene is low, some- compound. To simplify the process, a number of rapid and rela-
times less than one in 100 million cells. By increasing the tively inexpensive tests have been developed to screen for pos-
proportion of mutant cells, it is easier to isolate them. sible carcinogens. All examine the mutagenic effect of the
Penicillin enrichment relies on the fact that penicillin kills chemical in a microbiological system. The Ames test, devised
only growing cells. The mutagenized cells are incubated by Bruce Ames and his colleagues in the 1960s, illustrates the
in glucose-salts broth containing penicillin. Prototrophs can concept of such tests. It relies on the fact that mutagens increase
multiply in this medium so most are killed, whereas the the frequency of spontaneous reversions.
non-multiplying auxotrophs survive (figure 8.15). The enzyme The Ames test uses direct selection to determine the effect of
a test chemical on the reversion rate of a histidine-requiring
auxotroph of Salmonella (figure  8.16). If the chemical is muta-
genic, the reversion rate will increase relative to that observed
when no chemical is added (the control). The test also gives some
Cells growing in idea about how hazardous the chemical may be by the number of
glucose-salts revertants that arise.
broth Many chemicals are not carcinogenic themselves but are con-
Penicillin kills actively verted to carcinogens by enzymatic reactions in animals. Bacterial
Auxotroph growing cells, so most cells may not produce these enzymes, so an extract of ground-up
prototrophs die;
Prototroph auxotrophs survive
rat liver—which has the enzymes to carry out these conversions—
Add penicillin
and incubate. because they cannot is also added to the suspected mutagen.
grow in glucose-salts Additional testing must be done on any mutagenic chemical to
broth.
determine if it is carcinogenic. Although data are not available on
the percentage of mutagens that are carcinogens, it is clear that the
Plate on Add penicillinase to Ames test is useful as a rapid screening test. So far, only compounds
nutrient agar. degrade penicillin. that increase the reversion rate have been shown to be carcinogenic.
Plate on nutrient agar.

MicroByte
In 2010, a government advisory panel concluded that environmental
pollutants are greatly underestimated as a cause of cancer.
Auxotroph
Prototroph
MicroAssessment 8.5
Mutants can be selected using either direct or indirect techniques.
Replica plating is used for indirect selection, sometimes preceded by
penicillin enrichment. The Ames test is used to screen chemicals to
Many colonies grow; most Few colonies grow; most determine which ones are possible carcinogens.
are prototrophs (wild type). are auxotrophic mutants.
13. Distinguish between the kinds of mutants that can be isolated by
direct and indirect selection.
FIGURE 8.15 Penicillin Enrichment of Mutants 14. How does penicillin enrichment make it easier to isolate mutants?
? Penicillin causes the growing cells to lyse, releasing all of their 15. How could you demonstrate by replica plating that the
contents into the medium. How does this complicate penicillin environment selects but does not mutate genes in bacteria? +
enrichment?
200 Chapter 8 Bacterial Genetics

Control plate Test plates

Bacteria requiring Bacteria requiring

histidine for histidine for
growth (His–) growth (His–)

Glucose-salts Glucose-salts
agar agar
(lacks histidine)

Liquid without
suspected Liquid containing
mutagen suspected mutagen

Incubation Incubation

Spontaneous His+ Many His+ revertant If chemical is If chemical is Most remain His–
revertants colonies a mutagen not a mutagen and do not grow

Few colonies form. Many colonies form. Few colonies form.

FIGURE 8.16 Ames Test to Screen for Mutagens A control plate determines the spontaneous rate of reversion to His+. If the chemical
tested is a mutagen, it will increase the rate of His+ reversion mutations.
? Why do some cells grow on the control plate in the absence of a mutagen?

HORIZONTAL GENE TRANSFER AS A MECHANISM

OF GENETIC CHANGE
Microorganisms commonly acquire genes from other cells, the mutants are useful. These differences make it possible to deter-
process of horizontal gene transfer. The movement of DNA from mine if the recipients have indeed acquired new characteristics.
one cell (the donor) to another (the recipient) is largely responsible The resulting cells, recombinants, have properties of each of the
for the rapid spread of antibiotic resistance, such as described for original strains.
Staphylococcus aureus earlier in this chapter. Figure  8.17 illustrates how horizontal gene transfer can be
Horizontal gene transfer can be studied only if donor and demonstrated. Two bacterial strains—neither of which can grow
recipient cells are genetically different, which is one reason why on a glucose-salts medium because of multiple growth factor
 Part I Life and Death of Microorganisms 201

TABLE 8.3 Mechanisms of DNA Transfer

Strain A: Strain B:
His– Leu– Sensitivity
Trp– Mixture Thr– Main Size of DNA to DNase
Mechanism Feature Transferred Addition*
Transformation Naked DNA About 20 Yes
transferred genes

Transduction DNA Small fraction No

enclosed in a of the
bacteriophage chromosome
coat

Conjugation
Plasmid Cell-to-cell Entire plasmid No
transfer contact
required

Chromosome Cell-to-cell Variable No

transfer contact fraction of
Glucose-salts agar Glucose-salts agar required; only chromosome
Hfr cells can
be donors

Control: No His+ Trp+ Control: No Leu+ Thr+ *DNase is an abbreviation of deoxyribonuclease, an enzyme that degrades
mutants present mutants present DNA.

Genes can be transferred from a donor to a recipient by three

different mechanisms (table 8.3):
Glucose-salts
agar
1. DNA-mediated transformation: “Naked” DNA is taken up
by a cell.
Only prototrophic
recombinants (His+, 2. Transduction: A virus injects bacterial DNA into a cell.
Trp+, Leu+, Thr+) grow
3. Conjugation: DNA is transferred during cell-to-cell contact.

FIGURE 8.17 Experimental Demonstration of Horizontal Following gene transfer, recipient cells must replicate the DNA to
Gene Transfer in Bacteria Recombinant colonies exhibit genetic pass it on to daughter cells. This can happen only if the DNA is
traits from both strains present in the mixture. Control plates a replicon, meaning it has an origin of replication. Plasmids and
demonstrate that these colonies are not a result of spontaneous chromosomes are replicons, but fragments of chromosomal DNA are
mutation.
not. If a chromosomal fragment is transferred, then it must become
? When demonstrating horizontal gene transfer, why is it important part of a replicon to be maintained in a population (figure  8.18).
to use strains that each require at least two different amino acids? This involves a process called homologous recombination, which
can happen only if the donor DNA is similar in nucleotide sequence
to a region in the recipient cell’s genome. In homologous recombi-
nation, the donor DNA becomes positioned next to the comple-
mentary region of the recipient cell’s DNA. The donor DNA
then replaces a homologous segment of recipient DNA, and the
requirements—are used. Strain A is His– (requires histidine) and DNA it replaced is degraded. The molecular mechanisms involved
Trp– (requires tryptophan). Strain B is Leu– and Thr–, meaning that in homologous recombination are still not understood, and differ-
it requires leucine and threonine. Neither population is likely to ent mechanisms likely operate in different situations. origin of

give rise to a spontaneous mutant that grows on the glucose-salts replication, p. 166

agar because two simultaneous mutations in the same cell would

MicroByte
be required. The strains are mixed and then spread on a glucose- Fungal genes have been found in an aphid’s DNA, an extreme
salts agar plate. If colonies form, it suggests that cells of one strain example of horizontal gene transfer.
acquired genes from cells of the other strain.
202 Chapter 8 Bacterial Genetics

Bacterial DNA fragment

chromosome (no origin of replication)

DNA molecules without an origin

of replication cannot replicate in a cell.

Only one daughter cell

will have a copy of the
DNA fragment.

(a) Non-integrated DNA fragment

DNA fragment
(no origin of replication)

Homologous recombination

A DNA fragment integrated into a

bacterial chromosome can be replicated
and passed on to daughter cells.

All daughter cells

will have a copy of the
DNA fragment.

(b) Integrated DNA fragment

FIGURE 8.18 DNA Must Be Part of a Replicon to Be Maintained in a Population (a) DNA without an origin of replication will not be
passed on to any new daughter cells during growth of the population. (b) If the DNA becomes integrated within a replicon of the cell, it will be
inherited by all daughter cells.
? If the DNA fragment encodes penicillin resistance, how could you experimentally distinguish between (a) and (b)?

8.6 ■ DNA-Mediated DNA-mediated transformation, commonly referred to as transfor-

Transformation mation, involves the uptake of “naked” DNA by recipient cells
(see Perspective 8.1). Naked DNA is simply free in the cells’ sur-
Learning Outcome roundings; it is not contained within a cell or a virus. The fact that
the DNA is naked can be demonstrated by adding DNase (an
14. Describe the process of DNA-mediated transformation, including
the role of competent cells. enzyme that degrades DNA) to the medium. This prevents trans-
formation, indicating that the process requires naked DNA.
 Part I Life and Death of Microorganisms 203

PERSPECTIVE 8.1
The Biological Function of DNA: A Discovery Ahead of Its Time
In the 1930s, it was well known that DNA was MacLeod, and Maclyn McCarty, purified the although many scientists believe that they
present in all cells, including bacteria. Its func- active compound and then wrote one of the deserved it. Their studies pointed out that
tion, however, was a mystery. Since it consists most important papers ever published in biol- DNA is a key molecule in the scheme of life
of only four repeating subunits, most scientists ogy. In it, they reported that the transforming and led to James Watson and Francis Crick’s
did not believe it could be a very important molecule was DNA. determination of its structure, which they pub-
molecule. Its critical biological role was dis- The significance of their discovery was lished in 1953. The understanding of the struc-
covered through a series of experiments con- not appreciated at the time. Perhaps the dis- ture and function of DNA revolutionized the
ducted during a 20-year period by scientists in covery was premature, and scientists were study of biology and ushered in the era of
England and the United States. slow to recognize its significance. None of the molecular biology. Microbial genetics serves
In the 1920s, Frederick Griffith, an three investigators received a Nobel Prize, as its foundation.
English bacteriologist, was studying pneumo-
cocci, the bacteria that cause pneumonia. It
Organisms injected Results
was known that pneumococci could cause this
disease only if they made a polysaccharide
capsule (figure 1). In trying to understand the
Mouse dies
role of this capsule in causing disease, Griffith
injected living cells that could not synthesize
a capsule and heat-killed encapsulated cells Living encapsulated cells
into separate mice. As expected, the mice did
not develop pneumonia. He also injected
some mice with a mixture of heat-killed No effect
encapsulated cells and live non-encapsulated
cells. Much to his surprise, these mice devel-
Living non-encapsulated cells
oped pneumonia and died. Griffith isolated
living encapsulated pneumococci from the
dead mice.
Two years after Griffith reported these
findings, another investigator, M. H. Dawson, No effect
lysed heat-killed encapsulated pneumococci
and passed the suspension of ruptured cells Heat-killed encapsulated cells
through a very fine filter, through which only
the cytoplasmic contents of the bacteria could
pass. When he mixed the filtrate (the material
passing through the filter) with living bacteria
unable to make a capsule, some bacteria were Mouse dies
able to make a capsule. Moreover, the progeny Heat-killed encapsulated cells
of these bacteria could also make a capsule. +
Something in the filtrate was “transforming”
the harmless unencapsulated bacteria into ones
that could make a capsule.
Living non-encapsulated cells
What was this transforming principle? In
1944, after years of painstaking chemical
analysis of lysates capable of transforming Living encapsulated
cells isolated
pneumococci, three investigators from the
Rockefeller Institute, Oswald T. Avery, Colin FIGURE 1 Griffith’s Demonstration of Genetic Transformation

Naked DNA originates from cells that have either burst or take up DNA. Most competent bacteria take up DNA regardless of
secreted some DNA. When cells burst, the long chromosomes that its source. Some species accept DNA only from closely related
were tightly jammed into the cells break up into hundreds of bacteria, however, recognizing it by characteristic nucleotide
pieces. Some bacterial species secrete small pieces of DNA, pre- sequences located throughout the genome.
sumably as a means of promoting transformation. In the several dozen species that can become competent in
nature, the process is tightly controlled. Some species are always
competent, whereas others become so only under specific condi-
Competence tions, such as when the population reaches a critical density or under
In order for transformation to occur, the recipient cell must be certain nutritional conditions. The fact that some bacterial species
competent—a specific physiological state that allows the cell to become competent only under precise environmental conditions
204 Chapter 8 Bacterial Genetics

highlights the remarkable ability of these seemingly simple cells to

1 Gene conferring StrS Recipient chromosome
sense their surroundings and adjust their behavior accordingly.
In the case of Bacillus subtilis, a two-component regulatory Gene
system recognizes when the supply of nitrogen or carbon becomes conferring StrR
scarce in the environment and activates a set of genes required for
competence. Competence also requires a high concentration of Double-stranded DNA binds to the surface of a competent cell.
bacteria, a role of quorum sensing. Presumably, this ensures that
DNA in the medium will contact the competent bacteria. Even
under optimal conditions, however, only a fraction of the popula- 2
tion ever becomes competent. This means that seemingly identical
cells in a population exposed to the same environment can differ
in their physiological properties. two-component regulatory system,
p. 178 quorum sensing, p. 177
Single strand enters the cell; the other strand is degraded.

The Process of Transformation

3
Double-stranded DNA molecules bind to specific receptors on the
surface of competent cells (figure 8.19 1 ). 2 Only one strand
enters the cell, however, because nucleases at the cell surface
degrade the other strand. 3 Once the donor DNA is inside the
The strand integrates into the recipient cell’s genome by homologous
recipient cell, it integrates into the genome by homologous recom- recombination. The strand it replaced will be degraded.
bination; the strand it replaced will be degraded. 4 Because only
a single strand integrates, when the chromosome is replicated and
the cell divides, only one daughter cell will inherit donor DNA. 4 Streptomycin-sensitive Streptomycin-resistant
In the laboratory, DNA transformation is most easily detected daughter cell daughter cell
if the transformed cells can multiply under selective conditions in
which the non-transformed cells cannot. 5 For example, if the
donor cells are StrR and the recipient cells are StrS, then cells trans-
formed to StrR will grow and form colonies on a medium that
contains streptomycin. Although many other donor genes besides After replicating the DNA, the cell divides.
StrR will be transferred and incorporated by the recipient strain,
these transformants will go undetected without a mechanism to
recognize them. 5

MicroAssessment 8.6
In DNA-mediated transformation, DNA is released from donor cells
and taken up by competent recipient cells. Competent cells bind DNA Non-transformed cells (StrS) die on streptomycin-containing medium,
whereas transformed cells (StrR) can multiply.
and take up a single strand; that strand then integrates into the genome
by homologous recombination.
16. How does DNAse prevent transformation? FIGURE 8.19 DNA-Mediated Transformation The donor
17. Describe two ways by which DNA is released from cells. DNA in this case contains a gene conferring resistance to streptomycin
(StrR). Genes for resistance and sensitivity to streptomycin may differ
18. How could DNA-mediated transformation be used to test by only a single nucleotide.
chemicals for their mutagenic activity? +
? The cell at the top of this figure is competent. What does this
mean?

8.7 ■ Transduction which transfers only a few specific genes, will be described in
chapter 13—after the infection cycle of bacteriophages able to
Learning Outcome carry out this process is discussed.
15. Describe generalized transduction. To understand generalized transduction, you need to know
something about phages and how they infect bacterial cells. This
Bacterial viruses, called bacteriophages, or simply phages, can subject is covered more fully in chapter 13, so we will cover only
transfer bacterial genes from a donor to a recipient by transduction. the bare essentials here. Phages consist of genetic material, either
There are two types of transduction, generalized and specialized. In DNA or RNA (never both), surrounded by a protein coat. A phage
this chapter, we will describe only generalized transduction, which infects a bacterium by attaching to the cell and then injecting its
transfers any genes of the donor cell. Specialized transduction, nucleic acid. Enzymes encoded by the phage genome then cut the
 Part I Life and Death of Microorganisms 205

bacterial DNA into small pieces. Next, the cell’s enzymes repli- and inject the DNA it contains. The transducing particle, however,
cate the phage nucleic acid and synthesize proteins that make up injects bacterial DNA. The bacterial DNA may then integrate into
the empty phage coat. The phage nucleic acid then enters the the host chromosome by homologous recombination.
phage coat, and the various components assemble to produce com-
plete phage particles. These are then released, usually as a result
of host cell lysis. The phage particles then attach to other bacterial MicroAssessment 8.7
cells and begin new cycles of infection. Transduction results from an error that occurs during the infection
Generalized transduction results from a rare error that cycle of bacteriophages, leading to the transfer of bacterial genes from
sometimes occurs during the construction of phage particles one bacterial cell to another.
(figure 8.20). A fragment of bacterial DNA—produced when the 19. What error leads to generalized transduction?
phage-encoded enzyme cuts the bacterial genome—mistakenly 20. What is a transducing particle?
enters the protein coat. The product is called a transducing particle; 21. Two bacterial genes are transduced simultaneously. What does this
it carries no phage DNA and therefore is not a phage. Like phage suggest about the location of the two genes relative to each other? +
particles, a transducing particle will attach to another bacterial cell

Phage Transducing particle

1 A bacteriophage attaches to a 1 A transducing particle

specific receptor on a host cell. attaches to a specific
receptor on a host cell.

Bacterial
2 The phage DNA enters the cell. DNA 2 The bacterial DNA is
The empty phage coat remains injected into a cell.
on the outside of the bacterium.

3 Enzymes encoded by the Replaced 3 The injected bacterial

phage genome cut the bacterial host DNA DNA integrates into the
DNA into small pieces. chromosome by
homologous recombination.

4 Phage nucleic acid is replicated

and coat proteins synthesized.

4 Bacteria multiply with new genetic material.

5 During construction of viral Replaced host DNA is degraded.
particles, bacterial DNA can
mistakenly enter a protein coat.
This creates a transducing (b) The process of transduction
particle that carries bacterial
Transducing DNA instead of phage DNA.
particle FIGURE 8.20 Generalized Transduction (a) An error
during construction of phage particles produces a transducing
particle, which contains bacterial DNA instead of phage DNA.
(b) The bacterial DNA carried by the transducing particle is injected
(a) Formation of a transducing particle into a new host, resulting in generalized transduction. Essentially any
bacterial gene can be transferred this way.

? After a phage injects its DNA into a bacterial cell, the cell begins making proteins that make up the phage coat. Why does the same thing not
happen when a generalized transducing particle injects the DNA it carries?
206 Chapter 8 Bacterial Genetics

8.8 ■ Conjugation 3 Transferring DNA. A single strand of the F plasmid enters

the F – cell, passing through the F pilus. Once inside the
Learning Outcome recipient cell, that strand serves as a template for synthe-
16. Compare and contrast conjugation involving an F+ donor, an Hfr sis of the complementary strand, generating an F plasmid.
strain, and an F' donor. Likewise, the strand that remains in the donor serves as a
template for DNA synthesis, regenerating the F plasmid.
Conjugation is a complex process that requires contact between The transfer takes only a few minutes.
donor and recipient cells. Gram-positive and Gram-negative bac- 4 Transfer complete. Both the donor and recipient cells are
teria can both transfer DNA this way, but the process is quite dif- now F+ so they can act as donors of the F plasmid.
ferent in the two groups. For simplicity, however, we will consider
conjugation only in the more intensely studied Gram-negative
bacteria. 1 Making contact
Chromosome F plasmid
Plasmid Transfer F pilus
Plasmids are most frequently transferred to other cells by conjuga-
tion. These DNA molecules are replicons, so they can be repli-
cated inside cells, independent of chromosomal replication. Origin of
Conjugative plasmids direct their own transfer from donor Donor cell F+ transfer Recipient cell F−
to recipient cells. The most thoroughly studied example is the
The F pilus contacts the recipient F− cell.
F plasmid (F stands for fertility) of E. coli. Although this plasmid
does not encode any notable characteristics except those required
for transfer, other conjugative plasmids encode resistance to cer-
tain antibiotics, which explains how such resistance can easily 2 Initiating transfer
spread among a population of cells. E. coli cells that harbor the
F plasmid are designated F+, whereas those that do not are F–. The
F plasmid encodes several proteins required for conjugation,
including the F pilus, also referred to as the sex pilus (figure 8.21).
sex pilus, p. 65 One strand is cut in
Plasmid transfer involves a series of steps (figure 8.22): the origin of transfer

1 Making contact. The F pilus of the donor cell binds to a The pilus retracts and pulls the donor and recipient cells together.
specific receptor on the cell wall of the recipient.
2 Initiating transfer. After contact, the F pilus retracts, pull-
ing the two cells together. Meanwhile, a plasmid-encoded 3 Transferring DNA
enzyme cuts one strand of the plasmid at a specific nucleo-
tide sequence, the origin of transfer.

F pilus
A single strand of the F plasmid is transferred to the recipient cell;
its complement is synthesized as it enters that cell. The strand
transferred by the donor is replaced, using the remaining strand
as a template for DNA synthesis.

4 Transfer complete

F+ cell
F+ cell
At the end of the transfer process, both the donor and recipient
2 μm cells are F+ and synthesize the F pilus.

FIGURE 8.21 F Pilus Joining a Donor and Recipient Cell FIGURE 8.22 Conjugation—F Plasmid Transfer
? What are the hairlike appendages on the cell on the left? ? How does the recipient cell change as a result of conjugation?
 Part I Life and Death of Microorganisms 207

Chromosome Transfer take approximately 100 minutes for this to occur, an unlikely
event because the connection between the two cells usually
Chromosomal DNA transfer is less common than plasmid
breaks sooner than that. Because the entire integrated F plasmid
transfer and involves Hfr cells (meaning high frequency of
is not transferred, the recipient remains F –. The transferred
recombination). These are strains in which the F plasmid has
chromosomal DNA is not a replicon, so it will be maintained
integrated into the chromosome by homologous recombination,
only if it integrates into the recipient’s chromosome through
which happens on rare occasions (figure 8.23). The transfer of
homologous recombination. Unincorporated DNA will be
chromosomal DNA involves the same general steps as transfer
degraded.
of the F plasmid. Like F+ cells, Hfr cells produce an F pilus, and
the F  plasmid DNA directs its transfer to the recipient cell.
Because the F plasmid DNA is integrated into the chromosome,
however, chromosomal DNA is also transferred, beginning with 1 Making contact Origin of
the genes on one side of the origin of transfer (figure 8.24). The transfer
Integrated
entire chromosome is generally not transferred because it would Chromosomal F plasmid Chromosome
genes
C a'
B
A b' c'
Formation of an Hfr cell

Chromosome F plasmid Donor cell Hfr Recipient cell F−

The F pilus contacts the recipient F− cell.

The F plasmid sometimes

F pilus integrates into the bacterial 2 Transferring DNA Origin of
F+ cell chromosome by homologous transfer
recombination, generating Origin of
an Hfr cell; the process transfer A B A a'
Integrated is reversible.
F plasmid C b' c'

A single strand of the donor chromosome begins to be

Hfr cell transferred, starting at the origin of transfer. Gene A, closest
to the origin, is transferred first. DNA synthesis creates
complementary strands in both cells.

Formation of an F' cell

3 Transfer ends
Integrated
F plasmid B
A C A a'
B
b' c'
Chromosome
Hfr cell An incorrect excision of the
integrated F plasmid brings
The donor and recipient cells separate, interrupting DNA transfer.
along a portion of the
Chromosomal F' plasmid chromosome, generating
DNA an F' cell.
4 Integration of transferred DNA
F pilus
B
F' cell A C A

B c'
FIGURE 8.23 Formation of Hfr and F’ Cells An Hfr cell is
created when the plasmid integrates into the chromosome as a result Hfr cell
of recombination between homologous regions on the F plasmid and F− cell
chromosome. Note that the process is reversible, as recombination The donor DNA is integrated into the recipient cell’s chromosome
between the same two regions will excise the F plasmid, changing by homologous recombination. Unincorporated DNA is degraded.
the Hfr cell back to an F+ cell. An F’ cell is created when certain The recipient cell is still F–.
recombination events result in an incorrect excision that removes a
piece of the chromosome along with the F plasmid. This process is
also reversible. FIGURE 8.24 Conjugation—Chromosomal DNA Transfer
? Why does the F plasmid integrate only at specific locations? ? Why is the entire donor chromosome seldom transferred?
208 Chapter 8 Bacterial Genetics

F´ Donors different strains, make up the mobile gene pool or mobilome. These
genes can move from one DNA molecule to another, carried on
Hfr strains can revert to F+ because the process of F plasmid inte-
mobile genetic elements including various plasmids, transposons,
gration is reversible (see figure 8.23). In some instances, however,
regions called genomic islands, and phage DNA (table  8.4).
an error occurs during excision, and a piece of the bacterial chro-
Surprisingly, when all the non-conserved genome components of all
mosome is removed along with the F plasmid DNA. This action
the various E. coli strains are considered as a group, these sequences
brings a chromosomal fragment into the F plasmid, producing a
vastly outnumber sequences in the core genome.
plasmid called F' (F prime). Like the F plasmid, F' is a replicon
that is rapidly and efficiently transferred to F – cells. In the case of
F', however, the chromosomal fragment is transferred as well. Plasmids
Plasmids are common in the microbial world and are found in
MicroAssessment 8.8 most members of the Bacteria and Archaea and in some Eucarya.
Conjugation requires contact between donor and recipient cells. A Like chromosomes, most plasmids are double-stranded DNA mol-
donor cell that synthesizes an F pilus transfers the DNA to one that ecules. They have an origin of replication and therefore can be
does not. Both plasmid and chromosomal DNA can be transferred. replicated by the cell before it divides, but they replicate indepen-
Following transfer, plasmids replicate, but chromosomal DNA must dently of the chromosome. Plasmids generally do not encode
be integrated into a replicon. information essential to the life of a cell, and therefore cells can
22. The F plasmid encodes which two functions essential for survive their loss (curing). They are important, however, because
conjugation? they provide cells with the ability to cope in a particular environ-
23. Describe the outcomes of the three types of matings ment (table 8.5). plasmid, p. 66
(F+ × F –, Hfr × F –, and F' × F –). Plasmids vary with respect to their properties. Some carry
24. Would you expect transfer of chromosomal DNA by conjugation only a few genes, others carry a thousand. Low-copy-number
to be more efficient if cells were plated together on solid plasmids occur in only one or a few copies per cell, whereas high-
medium (agar) or mixed together in a liquid in a shaking copy-number plasmids are present in many copies, perhaps 500.
flask? Explain. + Most plasmids have a narrow host range, meaning they can repli-
cate in only one species. Broad host range plasmids replicate in
many different species, sometimes including both Gram-negative
8.9 ■ The Mobile Gene Pool and Gram-positive bacteria. Some plasmids cannot be maintained
in the same cell. Because of this, scientists have arranged them
Learning Outcomes into different compatibility groups—members of the same com-
17. Describe how plasmids differ from bacterial chromosomes. patibility group cannot be maintained in the same cell.
18. Compare and contrast transposons and genomic islands. Many bacterial plasmids are readily transferred by conjugation.
Conjugative plasmids carry all of the genetic information needed
Advances in genomics have uncovered surprising variation in the for transfer, including an origin of transfer. In contrast, mobilizable
gene pool (the sum of all genes) of even a single species. For plasmids encode an origin of transfer but lack other genetic infor-
example, nucleotide sequence analysis of many E. coli strains indi- mation required for transfer. However, when a conjugative plasmid
cates that only about 75% of a strain’s genes are found in all strains is in the same cell as a mobilizable plasmid, both plasmids can be
of that species. These conserved genes make up the core genome of transferred. Some plasmids can transfer between unrelated species
the species. The remaining ones, which vary considerably among and even between Gram-positive and Gram-negative bacteria. The

TABLE 8.4 The Mobile Gene Pool

Composition Property

Transposons
Insertion sequences Transposase gene flanked by short repeat Move to different locations in DNA in same cell
(ISs) sequences
Composite transposons Recognizable gene flanked by insertion Same as insertion sequences, but encode additional
sequences information
Plasmids Circular double-stranded DNA replicon; smaller Generally code only for non-essential genetic information
than chromosomes
Genomic Islands Large fragment of DNA in a chromosome or Code for genes that allow cell to occupy specific
plasmid environmental locations
Phage DNA Phage genome May encode proteins important to bacteria
 Part I Life and Death of Microorganisms 209

R Plasmid
TABLE 8.5 Some Plasmid-Coded Traits
Carbenicillin-resistance
Organisms in Which gene Chloramphenicol-
Trait Trait Is Found resistance gene

Tetracycline-
Antibiotic resistance Many, including Escherichia coli,
resistance
Salmonella sp., Neisseria sp., gene
Staphylococcus sp., and
Shigella sp.
Pilus synthesis E. coli, Pseudomonas sp.
Tumor formation in plants Agrobacterium sp. Origin of Pilus-synthesis
(see Perspective 8.2) replication genes
Nitrogen fixation Rhizobium sp.
Oil degradation Pseudomonas sp. Origin of transfer

Gas vacuole production Halobacterium sp. FIGURE 8.25 Functional Regions of an R Plasmid Genes
Insect toxin synthesis Bacillus thuringiensis conferring resistance to antimicrobial compounds tend to be clustered
within a specific region (the top half of the illustrated plasmid), whereas
Plant hormone synthesis Pseudomonas sp.
genes involved in replication and conjugative transfer are located on a
Antibiotic synthesis Streptomyces sp. different region (the bottom half of the illustrated plasmid).
Increased virulence Yersinia sp. ? How is an R plasmid similar to an F plasmid?
Toxin production Bacillus anthracis
the plasmids can give rise to a wide range of organisms becoming
resistant to many different antimicrobials. Members of the nor-
genes of an unusual plasmid can even be transferred into plant cells mal microbiota can harbor R plasmids, and then transfer them to
by conjugation (see Perspective 8.2). pathogens. normal microbiota, p. 8

Resistance Plasmids Transposons

Resistance, or R, plasmids encode resistance to many different In addition to causing mutations, transposons provide a mechanism
antimicrobial medications and heavy metals, such as mercury and for transferring various genes. Transposons can move into other
arsenic, all of which are found in hospital environments. Many of replicons in the same cell without any specificity as to where they
these plasmids are conjugative, composed of two parts: the resis- insert.
tance genes (R genes), which encode the resistance traits, and a Several types of transposons exist, varying in their structural
resistance transfer factor (RTF), which encodes the properties complexity. The simplest, an insertion sequence (IS), encodes
required for conjugation (figure 8.25). only the enzyme responsible for transposition, called transposase
R plasmids can give simultaneous resistance to numerous anti- (figure 8.26). Flanking the gene are inverted repeats—sequences
microbials, and many have a broad host range. As a consequence, that are identical when read in the 5' to 3' direction.

FIGURE 8.26 Transposable Elements The borders

Insertion sequence of insertion sequences are defined by inverted repeats 15–20
nucleotides in length. The first six nucleotides are shown here
Mobile element in expanded view to demonstrate their inverted orientation.
Composite transposons consist of two IS elements and the DNA
between them, all of which move as a single unit.
Transposase gene ? Why are some transposons medically important?
Inverted repeat Inverted repeat

5' 3' 5' 3'

3
T C G A T G... ...C A T C G A
A G C T A C... ...G T A G C T
3' 5' 3' 5'

Composite transposon

Mobile element

Insertion sequence Antibiotic-resistance gene Insertion sequence

210 Chapter 8 Bacterial Genetics

PERSPECTIVE 8.2
Bacteria Can Conjugate with Plants: A Natural Case of Genetic Engineering
For more than 50 years, scientists have known it is not needed to maintain the changes to source of carbon and energy, whereas most
that DNA can be transferred between bacteria. the plant cells. other bacteria in the soil, as well as plants, can-
Thirty years ago, it was shown that a bacte- The explanation of how Agrobacterium not. In other words, A. tumefaciens alters the
rium can even transfer its genes into cells of tumefaciens causes crown gall tumors was metabolism of the plant to produce food that
plants, including tobacco, carrots, and cedar established in 1977 following a report that only Agrobacterium cells can use. Thus,
trees, through a process analogous to conjuga- all crown gall–causing strains contain a large Agrobacterium tumefaciens is a natural genetic
tion. What led to this discovery started about plasmid termed the Ti (tumor-inducing) engineer of plants.
100 years ago in the laboratory of a plant plasmid. A group of microbiologists then The Agrobacterium–crown gall system is
pathologist, Dr. Erwin Smith. He showed that showed that a specific piece of the Ti plasmid, important for several reasons. First, it shows
the agent that causes a common plant disease, called T-DNA (transferred DNA) moves from that DNA can be transferred from prokaryotes
crown gall, is a bacterium—Agrobacterium the bacterial cell to the plant cell, where it to eukaryotes. Many people believed that such
tumefaciens. becomes incorporated into the plant chromo- transfer would be impossible in nature and
Crown gall is characterized by large galls some (figure  1). Because no regions of the could occur only in the laboratory. Second, this
or tumors that occur on the plant at the site of plant DNA are similar to those of bacteria, system has spawned an industry of plant bio-
infection, usually near the soil line, the crown integration occurs through non-homologous technology dedicated to improving the quality
of the plant. When other investigators cultured recombination. Like conjugation between bac- of higher plants. With this technology, it is pos-
the diseased plant tissue, they found it had teria, a pilus is required for DNA transfer. sible to replace the genes of hormone and
properties that differed from normal plant tis- Once incorporated into the plant chromo- opine synthesis in the Ti plasmid with any
sue. Whereas normal tissue requires several some, the T-DNA provides the plant cell with other genes, which will then be transferred and
plant hormones for growth, crown gall tissue additional genetic information, thereby giving incorporated into the plant. Examples of bene-
grows in the absence of these hormones. In it new properties. The promoters in T-DNA ficial genes that have been transferred into a
addition, crown gall tissue synthesizes large resemble those of plants rather than those wide variety of different plants include those
amounts of a compound, an opine, that neither of bacteria, so the genetic information is conferring resistance to viral pathogens,
normal plant tissue nor A. tumefaciens synthe- expressed in plants but not in A. tumefaciens. insects, and different herbicides. Rice has
sizes. Scientists learned that T-DNA encodes been engineered to synthesize high levels of
What surprised scientists studying the enzymes for the synthesis of the plant hor- β-carotene, the precursor of vitamin A.
Agrobacterium–crown gall system was mones as well as for the opine. The expression Genetic engineering of plants became a
the observation that the plant cells main- of these genes supplies the plant cells with the reality once scientists learned how a common
tained their altered growth characteristics plant hormones—explaining why the plant soil bacterium caused a well-recognized and
and the ability to synthesize opine even cells can grow in the absence of added hor- serious plant disease. This system serves as an
after the bacteria were killed by penicillin. mones—and allows them to synthesize the excellent example of how solving a riddle in
Investigators concluded that the crown gall opine. promoter, p. 168 basic science can lead to major industrial
cells are permanently altered. Although A. Why does A. tumefaciens alter plant cells? applications.
tumefaciens is required to start the infection, This bacterium is able to use the opine as a

Figure 1 Agrobacterium tumefaciens Causes Crown Gall

by Transferring DNA to Plant Cells

Chromosome T-DNA Plant tumor tissue

T-DNA
Chloroplast
Plant DNA

Plant cell
nucleus
Ti plasmid
Agar
medium

Agrobacterium Agrobacterium cells Crown gall tumor Bacteria-free tumor Plant tumor cell
tumefaciens cell inoculated into stem develops at site tissue is hormone contains T-DNA
of plant of inoculation. independent and integrated into the
synthesizes an opine. plant chromosome.
 Part I Life and Death of Microorganisms 211

Composite transposons consist of one or more genes flanked with a strain of S. aureus susceptible to vancomycin, but resistant to
by ISs (figure 8.26). Like insertion sequences, they can move in many other antibiotics. After treatment with vancomycin, S. aureus
the same replicon or from one replicon to another in the cell. Their cells resistant to that drug were isolated from the patient. The only
movement is easily followed if they encode a recognizable gene obvious difference between the sensitive and resistance isolates was
product such as antibiotic resistance. They integrate into their new that the latter had a gene that encoded resistance to vancomycin,
location through non-homologous recombination, a process that inserted into a plasmid present in both strains. But where did the
does not require a similar nucleotide sequence in the region of resistance gene come from? Further analysis suggested that it was
recombination. The transposon simply inserts into a stretch of acquired from a vancomycin-resistant strain of Enterococcus
DNA; it does not replace the existing sequences. If a composite faecalis isolated from the same patient. The resistance gene in that
transposon inserts into a conjugative plasmid, it can then be trans- bacterium was identical to the one in the S. aureus isolate. It was
ferred to other cells. Any gene or group of genes can move to also part of a transposon integrated into a plasmid, but the plasmid
another site if flanked by ISs, but transposons that carry genes for was different from that in S. aureus. It appears that E. faecalis trans-
antibiotic resistance are particularly important medically. ferred its transposon-containing plasmid to the sensitive S. aureus
The introduction in this chapter presented a real-life example of by conjugation. This entering plasmid was apparently destroyed by
how a transposon allowed a strain of Staphylococcus aureus to enzymes in S. aureus, but before that happened, the transposon
become resistant to vancomycin. We now know how the resistant jumped to the plasmid already in the S. aureus cell. Considering the
strain likely arose (figure 8.27). The patient initially was infected ease with which DNA can move among cells in a bacterial popula-
tion, is it any wonder that antibiotic resistance is such a serious
problem in treating infectious diseases today?

Vancomycin- Plasmid
resistance gene Genomic Islands
(encoded on a
transposon Genomic islands are large DNA segments in a cell’s genome that
on a plasmid) originated in other species. This conclusion is based on the fact
that their nucleotide composition is quite different from the rest of
the cell’s genome. In general, each bacterial species has a charac-
Staphylococcus aureus teristic proportion of G-C base pairs, so a large segment of DNA
sensitive to vancomycin that has a very different G-C ratio suggests the segment originated
from a foreign source and was transferred to the cell through
Enterococcus faecalis horizontal gene transfer. GC content, p. 252
plasmid transferred
Enterococcus faecalis by conjugation
The characteristics encoded by genomic islands include utili-
resistant to vancomycin zation of specific energy sources, acid tolerance, development of
symbiosis, and ability to cause disease. Genomic islands that
Transposon encode the latter are called pathogenicity islands.
jumps from
one plasmid
to another. MicroByte
Almost 1,400 genes of the pathogen E. coli O157:H7 are not in the
laboratory strain E. coli K-12.
Plasmid from
Enterococcus
faecalis is MicroAssessment 8.9
destroyed.
Plasmids vary in size, copy number, host range, genetic composition,
compatibility to coexist with other plasmids, and their ability to
be transferred to other cells. One type of important plasmid is the
R plasmid, which codes for resistance to various antimicrobial
medications and heavy metals. Transposons can move from one
location to another in the same replicon or to other replicons.
Genomic islands are large DNA segments thought to have originated
in other species.
Vancomycin-resistant
Staphylococcus aureus
25. What functions must a plasmid encode to be self-transmissible?
26. What characteristic of a genomic island suggests that it
originated in another species?
FIGURE 8.27 Transfer of Vancomycin Resistance from
Enterococcus faecalis to Staphylococcus aureus Transfer of 27. What does the phrase “reservoir for R plasmids” mean
resistance involved both a plasmid and a transposon. when referring to plasmids carried by non-disease-causing
bacteria? +
? What role did the transposon play in the transfer of resistance?
212 Chapter 8 Bacterial Genetics

FUTURE CHALLENGES 8.1

Hunting for Magic Bullets
Because of the increasing resistance of micro- of the nucleotide sequence of DNA is called in the host are potential targets. It should be
organisms to current antimicrobial medica- genomics. The next step is to identify genes possible to design a protein that inhibits the
tions, the demand for new antimicrobials that necessary for the survival of the microorgan- virulence protein and thereby prevents disease.
will kill these resistant organisms is rapidly ism or required to cause disease. To gain some To develop an antimicrobial that inhibits
increasing. It is surprising and sobering to understanding of the function of any gene, an enzyme required for virulence requires a
realize that only two new classes of antimi- one must compare its DNA sequence with the great deal of information about the enzyme,
crobials have been introduced into clinics in sequence of all other genes that have been put how it folds, what its three-dimensional struc-
the past 40 years. The great challenge is to into a database, called GenBank. If the gene ture is, and whether or not it interacts with
develop new antimicrobial agents that strike at that has been sequenced is similar in nucleo- other proteins.
targets different from those attacked by current tide sequence to any other gene, then it is In the past, the search for antimicrobial
antimicrobials. assumed that the two genes have similar func- medications has relied on random screening.
With the revolution that has been occur- tions. Thus, if the function of a gene that has Scientists looked for growth inhibition of a
ring in biology over the past 15 years, the been sequenced in any organism is known, the pathogen by a large number of organisms
development of new antimicrobial agents may function of all genes with a similar sequence is isolated from soil samples collected from
become a reality. Promising new strategies are likely to be similar. This is the science of bio- around the world. Today, new technologies
based on knowing the genomic sequence of informatics, which involves the analysis of the based on microbial genomics should identify
microbial pathogens. Many of the microbial nucleotide sequence of DNA in order to under- new targets and provide a rational approach to
genomes which have now been sequenced stand what it codes for. Genes that are required developing new antimicrobials.
are human pathogens. The study and analysis for virulence in the pathogen but are not found

Summary
8.1 ■ Genetic Change in Bacteria place of the usual nucleobases, and they have different hydrogen-bonding
The genotype of bacteria can change either through mutations or properties (figure  8.8). Intercalating agents insert into the double helix
horizontal gene transfer (figure  8.1). Bacteria are haploid, so any and push nucleotides apart, resulting in frameshift mutations.
changes in DNA can easily alter the phenotype.
Transposition
MUTATION AS A MECHANISM Transposons can be introduced intentionally into a cell in order to
inactivate genes.
OF GENETIC CHANGE
8.2 ■ Spontaneous Mutations Radiation
Spontaneous mutations occur as a result of normal cell processes. Ultraviolet irradiation results in thymine dimer formation (figure  8.9).
They are stable but occasionally revert back to the non-mutant form. The repair mechanism can cause mutations. X rays cause single- and
The chance that two spontaneous mutations will occur within the same double-strand breaks.
cell is the product of the individual mutation rates.
Base Substitution 8.4 ■ Repair of Damaged DNA (table 8.2)

Base substitutions occur during DNA synthesis (figure 8.2). They result Repair of Errors in Nucleotide Incorporation
in silent, missense, and nonsense mutations (figure 8.3). DNA polymerases have a proofreading function. Mismatch repair
Deletion or Addition of Nucleotides removes a portion of the strand that has a misincorporated nucleotide.
Deleting or adding one or two nucleotides causes a frameshift mutation, A new DNA strand is then synthesized (figure 8.10).
changing the reading frame of the encoded protein (figure 8.4). This often Repair of Modified Nucleobases in DNA
results in a shortened non-functional protein.
Specific glycosylases remove oxidized guanine or other modified
Transposons (Jumping Genes) nucleobases in DNA (figure 8.11).
Transposons can move from one location to another in a cell’s genome.
Repair of Thymine Dimers
The gene into which the transposon jumps is insertionally inactivated
by the event (figure 8.5). In photoreactivation, an enzyme uses the energy of light to break the
bonds of the thymine dimer (figure  8.12). In excision repair, the dam-
8.3 ■ Induced Mutations aged single-stranded segment is removed and replaced.
Induced mutations are caused by mutagens (table 8.1).
SOS Repair
Chemical Mutagens SOS repair is a last-ditch repair mechanism that uses a new DNA
Some chemicals modify nucleobases, altering their hydrogen-bonding polymerase that has no proofreading ability but can bypass the damaged
properties (figure  8.7). Base analogs can be mistakenly incorporated in DNA. Consequently, the newly synthesized DNA has many mutations.
 Part I Life and Death of Microorganisms 213

8.5 ■ Mutant Selection 8.8 ■ Conjugation

Conjugation requires cell-to-cell contact.
Direct Selection
Direct selection is used to obtain mutants that grow under conditions
Plasmid Transfer
in which the parent cell cannot. These mutants are easy to isolate
(figure 8.13). F+ cells synthesize an F pilus, encoded on an F plasmid; the F plasmid
is transferred from an F + to an F – cell through the pilus (figure 8.22).
Indirect Selection
Indirect selection uses replica plating to isolate an auxotroph from a Chromosome Transfer
prototrophic parent strain (figure 8.14). Penicillin enrichment increases
Hfr strains have the F plasmid integrated into the chromosome
the proportion of auxotrophic mutants in a culture (figure 8.15).
(figure  8.23). When the F plasmid is transferred, chromosomal DNA
Screening for Possible Carcinogens moves into a recipient cell along with it (figure 8.24).
The Ames test is used to determine if a chemical is a mutagen and
therefore a possible carcinogen (figure 8.16). F´ Donors
An F' donor carries a modified F plasmid that contains a piece of chro-
HORIZONTAL GENE TRANSFER AS mosomal DNA.
A MECHANISM OF GENETIC CHANGE (table 8.3)

For newly acquired DNA to replicate in a cell, it must either be a rep- 8.9 ■ The Mobile Gene Pool (table 8.4)
licon or integrate into the cell’s genome through homologous recombi- The mobile gene pool includes plasmids, transposons, genomic islands,
nation (figure 8.18). as well as phage DNA.

8.6 ■ DNA-Mediated Transformation Plasmids (table 8.5)

DNA-mediated transformation transfers “naked” DNA.
Plasmids are replicons that code for non-essential information; many
Competence are readily transferred by conjugation. R plasmids code for antibiotic
A cell must be competent to take up DNA, and certain species become resistance (figure 8.25).
competent in nature.
Transposons
The Process of Transformation
Transposons provide a mechanism for transferring genes. An insertion
Short strands of double-stranded DNA bind to cells, but only one strand
sequence (IS) encodes only transposase (figure  8.26). A composite
enters (figure 8.19).
transposon has one or more genes flanked by insertion sequences.
8.7 ■ Transduction
Transduction is the transfer of bacterial DNA by bacteriophage. Genomic Islands
There are two types, generalized transduction and specialized trans- Genomic islands are large DNA segments in a cell’s genome that origi-
duction (figure 8.20). nated in other species.

Review Questions
Short Answer Multiple Choice
1. How is an auxotroph different from a prototroph? 1. A culture of E. coli is irradiated with ultraviolet (UV) light.
Answer questions 1 and 2 based on this statement. The UV light
2. Why is deleting one nucleotide generally more detrimental than
specifically
deleting three?
a) joins the two strands of DNA together by covalent bonds.
3. What type of mutation in an operon is most likely to affect the
b) joins the two strands of DNA together by hydrogen bonds.
synthesis of more than one protein?
c) forms covalent bonds between thymine molecules on the same
4. What is meant by “proofreading” with respect to DNA poly- strand of DNA.
merase? d) forms covalent bonds between guanine and cytosine.
5. Why would a cell use SOS repair, considering that it introduces e) deletes bases.
mutations? 2. The highest frequency of mutations would be obtained if, after
6. Why is replica plating used to isolate an auxotrophic mutant from irradiation, the cells were immediately
a prototrophic parent? a) placed in the dark.
7. What is transduction? b) exposed to visible light.
c) shaken vigorously.
8. How is an F+ strain different from an Hfr strain?
d) incubated at a temperature below their optimum for growth.
9. Name four mobile genetic elements. e) The frequency would be the same no matter what the
10. Why are R plasmids important? environmental conditions are after irradiation.
214 Chapter 8 Bacterial Genetics

3. Penicillin enrichment of mutants works on the principle that 4. generalized transduction.

a) only Gram-positive cells are killed. 5. cell movement.
b) cells are most sensitive to antimicrobial medications during the a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5
lag phase of growth. 9. A plasmid that can replicate in E. coli and Pseudomonas is most
c) most Gram-negative cells are resistant to penicillin. likely a/an
d) penicillin kills only growing cells. a) broad host range plasmid.
e) penicillin inhibits formation of the lipopolysaccharide layer. b) self-transmissible plasmid.
4. Repair mechanisms that occur during DNA synthesis are c) high-copy-number plasmid.
1. mismatch repair. d) essential plasmid.
2. proofreading by DNA polymerase. e) low-copy-number plasmid.
3. light repair. 10. The frequency of transfer of an F' molecule by conjugation is clos-
4. SOS repair. est to the frequency of transfer of
5. excision repair. a) chromosomal genes by conjugation.
a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5 b) an F plasmid by conjugation.
5. You are trying to isolate a mutant of wild-type E. coli that requires c) an F plasmid by transformation.
histidine for growth. This can best be done using d) an F plasmid by transduction.
1. direct selection. e) an R plasmid by DNA transformation.
2. replica plating.
3. penicillin enrichment. Applications
4. a procedure for isolating conditional mutants. 1. Some bacteria may have higher mutation rates than others follow-
5. reversion. ing exposure to UV light. Discuss a reason why this might be the
a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5 case. What experiments could you do to determine whether this is
6. The properties that all plasmids share are that they a likely possibility?
1. all carry genes for antimicrobial resistance. 2. A pharmaceutical researcher is disturbed to discover that the major
2. are self-transmissible to other bacteria. ingredient of a new drug formulation causes frameshift mutations
3. always occur in multiple copies in the cells. in bacteria. What other information would the researcher want
4. code for non-essential functions. before looking for a substitute chemical?
5. replicate in the cells in which they are found.
a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5
Critical Thinking +
7. The addition of DNase to a mixture of donor and recipient cells 1. You have the choice of different kinds of mutants for use in the
will prevent gene transfer via Ames test to determine the frequency of reversion by suspected
carcinogens. You can choose a deletion, a point mutation, or a
a) DNA transformation.
frameshift mutation. Would it make any difference which one you
b) chromosome transfer by conjugation.
chose? Explain.
c) plasmid transfer by conjugation.
2. You have isolated a strain of E. coli that is resistant to penicillin,
d) generalized transduction.
streptomycin, chloramphenicol, and tetracycline. You also observe
8. An F pilus is essential for that when you mix this strain with cells of E. coli that are sensitive
1. DNA-mediated transformation. to the four antibiotics, they become resistant to streptomycin, peni-
2. chromosome transfer by conjugation. cillin, and chloramphenicol but remain sensitive to tetracycline.
3. plasmid transfer by conjugation. Explain what is going on.
9
Biotechnology and
Recombinant DNA
KEY TERMS
Colony Blotting Technique used (or other macromolecules) according
to determine which colonies on an to their size.
agar plate contain a given nucleotide Genetic Engineering Deliberately
sequence. altering an organism’s genetic
DNA Cloning Procedure in which information using in vitro
a fragment of DNA is inserted into a techniques.
vector and then transferred to another Polymerase Chain Reaction
cell, where it then replicates. (PCR) In vitro technique used to
DNA Microarray A probe-based duplicate a specific region of a DNA
technique used to study gene molecule, increasing the number of
expression patterns. copies exponentially.
DNA Probe Single-stranded piece Recombinant DNA
of DNA, tagged with a detectable Molecule DNA molecule created
marker, that is used to detect a by joining DNA fragments from two
complementary sequence. different sources.
DNA Sequencing Process of Restriction Enzyme Type of
determining the nucleotide sequence enzyme that recognizes a specific
of a DNA molecule. nucleotide sequence and then cuts
the DNA within or near that site.
Fluorescence in situ
Hybridization (FISH) Technique Vector DNA molecule, often a
used to detect a given nucleotide plasmid, that functions as a carrier
sequence within intact cells on a of cloned DNA.
microscope slide.
Gel Electrophoresis A procedure
Technicians working at a bench in a DNA laboratory. used to separate DNA fragments

A Glimpse of History
In 1976, Argentinean newspapers reported a violent shootout between only surviving relatives were aunts and grandmothers, and it is difficult
soldiers and the occupants of a house in suburban Buenos Aires, leaving to use chromosomal DNA to show genetic relatedness between a child
the five extremists inside dead. Conspicuously absent from those reports and such relatives. Dr. King decided to investigate mitochondrial DNA
were the identities of the “extremists”—a young couple and their three (mtDNA). This organelle DNA, unlike chromosomal DNA, is inherited
children, ages 6 years, 5 years, and 6 months. Over the next 7 years, only from the mother. A child will have the same nucleotide sequence of
similar scenarios recurred as the military junta that ruled Argentina killed mtDNA as his or her siblings, the mother and her siblings, as well as the
thousands of citizens it perceived as threats. This “Dirty War,” as it maternal grandmother.
came to be known, finally ended in 1983 with the collapse of the mili- By comparing the nucleotide sequences of mtDNA in different
tary junta and the election of a democratic government. The new leaders individuals, Dr. King was able to locate key positions that varied exten-
opened previously sealed records that confirmed what many had already sively among unrelated people, but were similar in maternal relatives.
suspected—more than 200 children survived the bloodshed and had in Dr. King’s technique, developed out of a desire to help reunite families
fact been kidnapped and placed with families in favor with the junta. victimized by war, has now found many uses. Today her lab, now at
Dr. Mary-Claire King was at the University of California at Berkeley the University of Washington, still uses molecular biology techniques
when she was enlisted to help in the effort to return the children to the for humanitarian efforts, identifying the remains of victims of atrocities
surviving members of their biological families. Dr. King and others rec- around the world.
ognized that DNA technology could be used for this important humani-
tarian cause. By analyzing certain DNA sequences, blood and tissue

B
iotechnology uses microbiological and biochemical techniques
samples from one individual can be distinguished from those of another.
to solve practical problems and produce useful products. In
These same principles can also be used to show that a particular child is
the progeny of a given set of parents. Because a person has two copies the past, this often meant labor-intensive searches for natu-
of each chromosome—one inherited from each parent—half of a child’s rally occurring mutants that had desirable characteristics. Today,
DNA will represent maternal sequences and the other half will represent recombinant DNA techniques have made it possible to geneti-
paternal traits. The case of the Argentinean children was complicated, cally alter organisms to give them more useful traits. Researchers
however, because most of the parents were dead or missing. Often, the can isolate genes from one organism, manipulate the purified DNA

215
216 Chapter 9 Biotechnology and Recombinant DNA

in vitro, and then transfer the genes into another organism. In fact,
EcoRI recognition
biotechnology is now nearly synonymous with genetic engineering, sequence
the process of deliberately altering an organism’s genetic informa-
tion using in vitro techniques.
Chromosomal DNA is
Since the advent of recombinant DNA techniques, a virtual digested with EcoRI,
G G C T G A A T T C G C T T
C C G A C T T A A G C G A A
toolbox of DNA technologies has been developed. The infor- generating restriction
mation and innovations these have generated impact society in fragments.
numerous ways—from agricultural practices and medical diag- Cohesive (“sticky”) ends
noses to evidence used in courtrooms. Table 9.1 summarizes
the applications of the DNA-based technologies described in
G G C T G A A T T C G C T T
this chapter. C C G A C T T A A G C G A A

(a)
9.1 ■ Fundamental Tools
Used in Biotechnology N N N N G A A T T C N N N N
N N N N C T T A A + G N N N N
Learning Outcome
1. Describe the role of restriction enzymes and gel electrophoresis in Complementary cohesive ends
biotechnology. can anneal.

Before exploring the application of biotechnology, it is helpful to A A T T C N N N N

N N N N G
understand some of the basic components of a molecular biologist’s N N N N C T T A A G N N N N
“tool kit.” As we describe these, it is important to remember that
diagrams focus on only one or a few DNA molecules to illustrate
DNA ligase forms a covalent
what is happening at a molecular level. In reality, scientists are bond between adjacent
often working with millions of molecules. nucleotides.

N N N N G A A T T C N N N N
N N N N C T T A A G N N N N
Restriction Enzymes
Restriction enzymes allow scientists to easily cut DNA into frag-
ments in a predictable and controllable manner (figure  9.1a). (b)
Each enzyme recognizes a specific 4 to 6 base-pair nucleotide
sequence (table  9.2). The recognition sequences are typically FIGURE 9.1 Action of Restriction Enzymes (a) Digesting
DNA with a restriction enzyme generates restriction fragments.
palindromes, meaning they are the same on both strands when
(b) Fragments that have complementary cohesive ends can anneal,
read in the 5 to 3 direction. The enzyme cuts each strand of regardless of their original source (N = nucleotide, meaning that it
DNA within or near that sequence, digesting the DNA to generate could be any of the four nucleobases as long as base-pairing rules are
restriction fragments. restriction enzymes, p. 315 followed).
The name of a particular restriction enzyme represents the ? How do restriction enzymes make it easier for scientists to create
bacterium from which it was first isolated. The first letter is the recombinant DNA molecules?

TABLE 9.1 Applications of DNA-Based Biotechnologies

Technology Applications

Genetic engineering Genetically engineered microorganisms are used to produce medically and commercially valuable proteins, to
produce specific DNA sequences, and as a tool for studying gene function and regulation.
DNA sequencing Once the nucleotide sequence of even part of an organism’s DNA has been determined, the information can be
used to decipher the amino acid sequence of the encoded proteins. It can also be used to determine how similar the
nucleotide sequence is to DNA from other organisms, which can give insights into genetic relatedness.
Polymerase chain The presence of a specific segment of DNA can be detected, and the size determined, in only a matter of hours. This
reaction (PCR) can be used in diagnosis of an infectious disease if DNA specific to a pathogen can be amplified. It is also used to
“fingerprint” DNA for forensic evidence.
Probe technologies Colony blots are used to detect colonies that contain a specific DNA sequence; fluorescence in situ hybridization
(FISH) is used to identify cells directly in a specimen; DNA microarrays are used to study gene expression.
 Part I Life and Death of Microorganisms 217

Researchers use restriction enzymes not only to cut DNA, but

Examples of Common
TABLE 9.2 Restriction Enzymes also to create recombinant DNA molecules, which are molecules
made by joining DNA from two different sources. This is possible
Recognition because many restriction enzymes produce a staggered cut in the
Sequence recognition sequence, resulting in ends with short overhangs of usu-
(arrows indicate ally four nucleotides (figure 9.1b). The overhangs are called sticky
Enzyme Microbial Source cleavage sites) ends, or cohesive ends, because they will anneal (form base pairs)
AluI Arthrobacter luteus ↓ with one another. Any two complementary cohesive ends can
5 A G C T 3 anneal, even those from two different organisms. The enzyme DNA
3 T C G A 5 ligase is used to form a covalent bond between adjacent nucleotides,
↑
joining the two molecules. Thus, if restriction enzymes are viewed
BamHI Bacillus ↓ as scissors that cut DNA into fragments, then DNA ligase is the glue
amyloliquefaciens H 5 G G A T C C 3
that pastes the fragments together. DNA ligase, p. 167
3 C C T A G G 5
↑

EcoRI Escherichia coli RY13 ↓ Gel Electrophoresis

5 G A A T T C 3
3 C T T A A G 5 Gel electrophoresis separates DNA fragments by size (figure 9.2).
↑ The DNA samples are added to wells in a gelatin-like substance,
usually either agarose (a highly purified form of agar) or poly-
acrylamide. A size standard—a mixture of DNA fragments of
first letter of the genus name, and the next two are from the species known sizes—is routinely put into a well of the same gel. The size
name. In the past, these were italicized, but that rule has been standard serves as a basis for comparison, allowing the researcher
dropped. Other numbers or letters designate the strain and order of to determine the sizes of the various DNA fragments in the
discovery. For example, EcoRI is from E. coli strain RY13. samples. The gel is then placed in an electrophoresis buffer (an

Sample Sample Sample Size

1 2 3 standard
Power
source

23 kb*
9.4 kb
6.6 kb
4.4 kb
2.3 kb
2.0 kb
_

Negatively
charged
electrode * Fragment sizes in standard
kb = 1,000 base pairs
+

Positively
charged
electrode

a Samples are added to wells in the gel. As the DNA moves b A size standard serves as the c
through the gel, long fragments are slowed in the tangle basis for determining the size of
of the gel matrix, whereas short fragments move more the other fragments.
quickly. This separates fragments according to their size.

FIGURE 9.2 Gel Electrophoresis (a) Gel electrophoresis separates DNA fragments according to size. (b) A size standard serves as a basis for
determining the size of the other fragments. (c) DNA on the gel is visible when stained with ethidium bromide and viewed with UV light; each
fluorescent band represents millions of molecules of a specific-sized fragment.
? How does gel electrophoresis separate DNA fragments according to size?
218 Chapter 9 Biotechnology and Recombinant DNA

electrically conductive solution) and subjected to an electrical cur-

Some Applications of
rent. DNA is negatively charged, so the fragments move toward TABLE 9.3
Genetic Engineering
the positively charged electrode. As the DNA moves through the
gel, long fragments are slowed in the tangle of the gel matrix, Example Use
whereas short fragments move more quickly. PROTEIN PRODUCTION
The DNA is not visible in the gel unless it is stained. To do
Pharmaceutical Proteins
this, the gel is immersed in a solution containing ethidium bro-
mide. This dye binds DNA and fluoresces when viewed with UV Alpha interferon Treating cancer and viral
infections
light (figure  9.2c). Each fluorescent band represents millions of
molecules of a specific-sized fragment of DNA. Erythropoietin Treating some types of anemia
Gel electrophoresis can also be used to separate other macro- Beta interferon Treating multiple sclerosis
molecules, specifically RNA and proteins, according to their size. Deoxyribonuclease Treating cystic fibrosis
The basic principles are similar to that just described, but the gel
Factor VIII Treating hemophilia
compositions differ.
Gamma interferon Treating cancer

MicroAssessment 9.1 Glucocerebrosidase Treating Gaucher disease

Growth hormone Treating dwarfism
Restriction enzymes recognize specific nucleotide sequences, and then
cut the DNA, generating restriction fragments. Gel electrophoresis is Insulin Treating diabetes
used to separate DNA fragments according to their size. Platelet derived growth Treating foot ulcers in diabetics
1. What is the importance of cohesive ends in genetic engineering? factor
2. How does gel electrophoresis separate different-sized DNA Streptokinase Dissolving blood clots
fragments? Tissue plasminogen Dissolving blood clots
3. What should a restriction enzyme isolated from Staphylo- activator
coccus aureus strain 3A be called? +
Vaccines
Hepatitis B Preventing hepatitis
HPV Preventing cervical cancer
9.2 ■ Applications of Genetic
Foot-and-mouth disease Preventing foot-and-mouth
Engineering disease in animals

Other Proteins
Learning Outcome
2. Describe the applications of genetically engineered bacteria and Bovine somatotropin Increasing milk production
plants. in cows
Chymosin Cheese-making
Genetic engineering brought biotechnology into a new era by Restriction enzymes Cutting DNA into fragments
providing a powerful tool for manipulating microorganisms for
medical, industrial, and research uses (table 9.3). Plants and ani- DNA PRODUCTION
mals can now be genetically engineered as well. DNA for study Determining nucleotide
sequences; obtaining DNA
Genetically Engineered Bacteria probes

Genetically engineered bacteria have a variety of uses, including RESEARCHING GENE FUNCTION AND REGULATION
protein production, DNA production, and a tool for research. In
Creating gene fusions Studying the conditions that
fact, much of the information described in this textbook was affect gene activity
revealed through research using genetically engineered bacteria.
Genetic engineering relies on DNA cloning, a process that TRANSGENIC PLANTS
involves using a restriction enzyme to cut purified DNA of one
Pest-resistant plants Insect-resistant corn, cotton, and
organism and then transferring a fragment of that DNA into a dif- potatoes
ferent organism (figure 9.3). As part of the process, the transferred
Herbicide-resistant plants Engineered plants (soybean,
DNA must replicate in the recipient in order to be passed to prog- cotton, corn) are not killed by
eny, thereby generating a population of cells that harbors copies of biodegradable herbicides used
the DNA fragment (see figure 8.18). Most DNA fragments, how- to kill weeds.
ever, will not contain an origin of replication and therefore will not Plants with improved Rice that produces vitamin A
replicate independently in a cell. In addition, they generally have nutritional value and iron
no similarity to chromosomal sequences, so they will not integrate Plants that function as Enable researchers to study
into the chromosome by homologous recombination. To generate a edible vaccines foods as possible vehicles for
cloned nucleotide sequence that will be replicated in a cell, the DNA edible vaccines
fragment can be inserted into a plasmid or other independently
 Part I Life and Death of Microorganisms 219

High-copy-
number vector Gene
+

Gene inserted
into vector
Isolate DNA.

Use a restriction enzyme to

generate fragments of DNA.

Cut the vector with the

same enzyme used to
cut the genomic DNA.
Join vector and fragment
to create recombinant
Linear molecule.
Vector vector
Multiple copies of
the gene produced

FIGURE 9.4 Cloning into a High-Copy-Number Vector When

a gene is inserted into a high-copy-number vector, multiple copies of
Introduce recombinant
molecule into new host. that gene will be present in a single cell, resulting in the synthesis of
many molecules of the encoded protein.
? Why must the gene be inserted into a vector for it to be cloned?
New host
in treating diabetes, was one of the first important pharmaceutical
proteins to be produced through genetic engineering. The original
commercial product was extracted from pancreatic glands of cattle
and pigs and sometimes caused allergic reactions. Once the gene
for human insulin was cloned into bacteria, microbes became the
major source of insulin. The product is safer and more economical
than the version extracted from animal tissues.
Another medically important use of genetically engineered
Protein production DNA production Research microorganisms is vaccine production. Vaccines protect against
Pharmaceutical proteins DNA sequencing Studying gene function
Vaccines and regulation disease by harmlessly exposing a person’s immune system to killed
Proteins used in industry or weakened forms of the pathogen, or to parts of the pathogen.
Although vaccines are generally composed of whole microbes, only
FIGURE 9.3 Cloning DNA specific proteins are necessary to immunize (induce protection)
? Why would it be easiest to use the same restriction enzyme to cut against the disease. The genes coding for these proteins can be
both the vector and the insert DNA? cloned into yeast or bacteria, allowing large amounts of the pro-
tein to be produced and then purified. This type of vaccine is cur-
replicating DNA molecule to form a recombinant molecule. The rently used to prevent hepatitis B and cervical cancer in humans
DNA molecule used as a carrier of the cloned DNA is called a vec- and foot-and-mouth disease of domestic animals. vaccines, p. 421
tor and is commonly a plasmid that has been genetically modified. One of the most widely used proteins made by genetically
DNA that has been incorporated into the vector is called an insert. engineered organisms is chymosin (rennin), an enzyme used
origin of replication, p. 166 homologous recombination, p. 201 in cheese production. It causes milk to coagulate and produces
If a gene coding for a valuable protein is inserted into a high- desirable changes in the characteristics of cheeses as they ripen.
copy-number vector, a bacterium that harbors the recombinant Traditionally, chymosin was obtained from the stomachs of calves,
molecule can make large amounts of that protein. This is because but genetically engineered bacteria are now the main source. The
each gene copy can be transcribed and translated (figure  9.4). microbial product is less expensive and more reliably available.
 high-copy-number plasmid, p. 208 Other proteins produced by genetically engineered microbes include
various restriction enzymes and bovine somatotropin, a growth
Protein Production hormone used to increase milk production in dairy cows.
A number of different pharmaceutical proteins are now produced Some scientists hope to create microorganisms that can be
by genetically engineered microorganisms. In the past, these pro- readily customized and used as miniature factories. With that goal
teins were extracted from live animal or cadaver tissues, which in mind, researchers made a variation of the Mycoplasma mycoides
made them expensive and limited in supply. Human insulin, used chromosome “from scratch,” assembling it from machine-made
220 Chapter 9 Biotechnology and Recombinant DNA

DNA segments. In 2010, they successfully used it to replace the Green fluorescent
existing chromosome of a different Mycoplasma species. The pro- protein (GFP) gene
(reporter gene)
cedure may pave the way for creating synthetic bacteria that can
produce a wide range of commercially valuable substances. GFP

MicroByte 1 GFP inserted

The insulin yield from a 2,000-liter culture of E. coli cells with into gene A.
a cloned insulin gene is the same as 1,600 pounds of pancreatic
glands!
Gene A
2 Expression of gene A
DNA Production observed through GFP
In many cases, a researcher is interested in obtaining readily available expression.
supplies of certain DNA fragments. By cloning a segment of DNA
GFP
into a well-characterized bacterium such as E. coli, that sequence can
then be used for study and further manipulation.
Human genes are often cloned into bacteria to make them easier to 3 Cells fluoresce when
the GFP gene is expressed,
study. A human cell contains an estimated 25,000 genes, whereas indicating that the regulatory
E. coli contains only 4,500 genes; thus, a human gene cloned into the elements of gene A are
bacterium on a high-copy-number vector represents a much higher turned on.
percentage of the total DNA in the recipient cell than in the original cell.
This makes it easier to isolate the DNA as well as the gene product.
Random samples of DNA from any environment can be
cloned into E. coli and then the nucleotide sequence determined.
Gene On Gene Off
By doing this, the genomic characteristics of some of the 99% of
Green fluorescent No fluorescence
bacteria that have not been grown in culture can be studied. This protein produced
“shotgun cloning” is the first step in metagenomics, the study of
the total genomes in a sample. metagenomics, p. 184 FIGURE 9.5 The Function of a Reporter Gene The regulatory
elements are not shown here.
Research ? Why is the green fluorescent protein useful as a reporter gene?
The function and regulation of genes can be studied with experi-
ments that involve cloning. For example, gene regulation can be other non-engineered plants, in place of more persistent alterna-
studied by creating a gene fusion—joining the gene of interest and tives. The herbicide can be applied throughout the growing
a reporter gene (figure 9.5). The reporter gene encodes a product season, so the soil can be tilled less frequently, preventing erosion.
that is easy to see, such as green fluorescent protein (GFP). This Plants with improved nutritional value are also being devel-
makes it relatively simple to detect gene expression and, in turn, oped. Genes that code for the synthesis of β-carotene, a precursor
determine the conditions that affect gene activity. of vitamin A, have been introduced into rice. The rice was also
engineered to provide more dietary iron. The diet of much of the
world’s population is deficient in these essential nutrients, so
Genetically Engineered Eukaryotes advances such as these could profoundly effect global health.
Yeasts can be genetically engineered to perform many of the func- Plants that serve as edible vaccines are also being developed.
tions described for bacteria. They serve as an important model for Researchers have successfully genetically engineered potatoes
gene function and regulation in eukaryotic cells. and rice to produce certain proteins from pathogens and have
Multicellular organisms can also be genetically engineered. A shown that the immune system responds to these, raising hopes for
plant or animal that harbors a cloned gene is transgenic. Transgenic an edible vaccine. Whether the immune response is strong enough
plants are of particular interest to microbiologists because their to protect against disease is still unclear.
development started with basic research studying Agrobacterium
tumefaciens; the Ti plasmid of this bacterium has now been geneti- MicroAssessment 9.2
cally manipulated so it can be used as a vector to deliver desirable
Genetically engineered bacteria can be used to produce a variety of
genes to plant cells (see Perspective 8.2). products, including medically and commercially important proteins,
Corn, cotton, and potatoes have been engineered to produce a vaccines, and DNA for study. They are also used as research tools.
biological insecticide called Bt toxin, which is naturally produced by Genetic engineering can be used to develop pest-resistant plants,
the bacterium Bacillus thuringiensis as it forms endospores. Unlike herbicide-resistant plants, and plants that produce more nutrients.
many chemically synthesized toxins, Bt toxin is toxic only to insects, 4. Name a disease treated with a protein produced by genetically
including their larvae. endospore, p. 67 engineered microorganisms.
Soybeans, cotton, and corn have been engineered to resist the 5. Describe the characteristics of two different transgenic plants.
effects of the herbicide glyphosate (Roundup). This allows grow- 6. Why would calves’ stomachs have chymosin (rennin)? +
ers to apply this biodegradable herbicide, which kills weeds and
 Part I Life and Death of Microorganisms 221

An approach frequently used to clone a specific gene is to make a

9.3 ■ Techniques Used in DNA library, a collection of clones that together contain the
Genetic Engineering entire genome (figure 9.6). This is made by cutting the DNA of
the organism being studied with restriction enzymes and then
Learning Outcomes cloning the entire set of restriction fragments into a population of
3. Describe how a DNA library is made. E. coli cells. Although each cell in the resulting population con-
4. Explain how introns are removed from eukaryotic genes. tains only one fragment of the genome, the entire genome is rep-
5. Describe the characteristics of a typical vector. resented in the population as a whole. Once a DNA library has
6. Explain how to obtain cells that harbor recombinant molecules. been prepared, colony blots (which will be described later) can be
used to determine which cells contain the gene of interest.

FIGURE 9.6 A DNA Library Each cell

contains one fragment of a given genome.
Bacterium X
? What enzyme is used to join the genomic
fragments to vector molecules?

Isolate the total DNA

from the organism.

Use restriction enzymes

to generate genomic
fragments.

A B C D
E F
Cut the vector with the G
same enzyme used to H
cut the genomic DNA.

Vector
molecules Join genomic fragments
to vectors.

A B C D
Recombinant
molecules
E F G H

Introduce the recombinant

molecules into E. coli cells.

B
A population of E. coli cells
that, together, contain all
A of the fragments of the
genome of bacterium X.
C

E
H
G
222 Chapter 9 Biotechnology and Recombinant DNA

This section will describe methods used to clone DNA in bac- of DNA, or cDNA, encodes the same protein as the original
terial cells. For information regarding the engineering of eukary- DNA  but lacks introns (figure  9.7). introns, p.  176 reverse
otic organisms, see Perspective 8.2 and visit the text website transcriptase, p. 320
(www.mhhe.com/nester7).
Generating a Recombinant
Obtaining DNA DNA Molecule
The first step of a cloning experiment is to obtain the DNA that As described earlier, restriction enzymes and DNA ligase are used
will be cloned. This is done by adding a detergent to cells in a to create recombinant molecules. The vector, usually a modified
broth culture to lyse them. As the cells burst, the relatively fragile plasmid or bacteriophage, has an origin of replication and func-
DNA is inevitably sheared into many pieces of varying lengths. tions as a carrier of the cloned DNA (figure  9.8). Vectors must
When cloning eukaryotic genes into bacteria for protein pro- also have at least one restriction enzyme recognition site. This
duction, a copy of DNA that lacks introns must first be obtained. allows the circular vector to be cut, forming a linear molecule to
To do this, mature mRNA is isolated from the appropriate tissue; which the insert can be joined. Many vectors have been engi-
recall that introns have been removed from these molecules. Then, neered to contain a multiple-cloning site, a short sequence that has
a strand of DNA complementary to the mRNA is synthesized the recognition sequences of several different restriction enzymes.
in vitro using reverse transcriptase, an enzyme from retroviruses. The value of a multiple-cloning site is its versatility—a fragment
That strand of DNA is then used as a template for synthesis of its obtained by digesting with any of a number of different restriction
complement, creating double-stranded DNA. The resulting copy enzymes can be inserted into the site.

Intron Intron

Eukaryotic DNA
Eukaryotic
cell DNA Transcription of eukaryotic
DNA generates pre-mRNA.

mRNA

Pre-mRNA Splicing removes the

introns from the pre-mRNA,
generating mRNA.

mRNA

mRNA extracted
from cells

mRNA
Isolated (isolated) Reverse transcriptase
mRNA synthesizes DNA from mRNA.

cDNA
Single strand
of cDNA DNA polymerase completes
the cDNA, free of introns.

Double-stranded
cDNA

FIGURE 9.7 Making cDNA from Eukaryotic mRNA In order for eukaryotic genes to be expressed by a prokaryotic cell, a copy of DNA
without introns must be cloned. The cDNA encodes the same protein as the original DNA but lacks introns.
? What enzyme is used to make cDNA from pre-mRNA?
 Part I Life and Death of Microorganisms 223

Origin of to form a blue compound. Because the multiple-cloning site of

replication pUC18 is within the lacZ gene, creation of a vector-insert hybrid
results in a non-functional gene product. Thus, cells that harbor
intact vector have a functional lacZ gene and form blue colonies,
Selectable Marker whereas those that contain a recombinant molecule form white
A gene encoding resistance ones. insertional inactivation, p. 192
to an antibiotic such as ampicillin
Various vectors are available for cloning eukaryotic
DNA into bacterial cells, and the choice depends largely
on the purpose of the procedure. If the goal is to produce
the encoded protein, then a vector designed to optimize
transcription and translation of the insert DNA is used.
To create a DNA library of a human or other eukaryotic
genome, a vector that can carry a large insert is generally
used. For more information about these vectors, visit the
text website (www.mhhe.com/nester7).

Obtaining Cells That Harbor

Recombinant DNA Molecules
Second Genetic Marker
A gene such as lacZ ' that
Once recombinant plasmids are generated, they must be trans-
TAC GAATTC CCC GGATCC GTCGAC C
ATG CTTAAG GGG CCTAGG CAGCTG G encodes an observable ferred into suitable hosts where the molecules can replicate. For
EcoRI BamHI SalI phenotype routine cloning experiments, one of the many well-characterized
laboratory strains of E. coli is generally used. These strains are
Multiple-Cloning Site
Contains the recognition sequence
of several different restriction enzymes
Vector Disrupted
lacZ' gene
FIGURE 9.8 Typical Properties of an Ideal Vector Most lacZ' gene
vectors have an origin of replication, a selectable marker, and a
multiple-cloning site. A second genetic marker, used to differentiate
cells containing recombinant plasmids from those that contain an
intact vector, spans the multiple-cloning site.
Insert
? When a vector has been successfully joined with a vector, will
the selectable marker still be functional? Will the second genetic
marker still be functional? mRNA mRNA

Non-functional
Vectors typically have a selectable marker, a gene that Functional lacZ' protein
allows cells to grow in otherwise inhibitory or lethal conditions. gene product
This is important because even under ideal conditions, most cells
in a population do not take up DNA. The selectable marker allows
the researcher to eliminate those cells that have not taken up either
the recombinant molecule or the vector. A common selectable Blue colonies White colonies
marker is the gene that encodes resistance to the antibiotic ampicil-
lin; cells that have taken up a vector or recombinant molecule are
able to grow on a medium containing ampicillin. ampicillin, p. 464
Most vectors have a second genetic marker, in addition to the
selectable marker, used to distinguish cells that contain recombinant
plasmids from those containing an intact vector. This is important
because when the vector and insert DNA are both cut with the same
restriction enzyme and the fragments mixed, not all will form the
desired recombinant molecules. For example, the two ends of the
vector can anneal to each other, regenerating the circular vector, Bacterial cells plated on
medium containing X-gal
which can replicate when introduced into a cell. The second genetic
marker is situated so that it will be insertionally inactivated when an FIGURE 9.9 The Function of the lacZ Gene in a Vector The
insert is in the multiple-cloning site. A good illustration of the utility lacZ gene is used to differentiate cells that contain recombinant
of the second genetic marker is provided by a vector called pUC18 plasmid from those that contain vector alone.
(figure 9.9). The second genetic marker of pUC18 is a gene called ? What color colonies will cells that harbor a recombinant molecule
lacZ. The product of this gene cleaves a colorless chemical, X-gal, form?
224 Chapter 9 Biotechnology and Recombinant DNA

easy to grow, and they are well characterized with respect to their information be used to deny an individual certain rights and privi-
genetics, biochemistry, and antibiotic sensitivities. leges? It is important that ongoing discussions about these complex
A common method of introducing DNA into a bacterial host issues continue as the technologies advance.
is DNA-mediated transformation. E. coli cells are not naturally Genetically modified (GM) organisms hold many promises,
competent and must be specially treated to induce them to take up but the debate over their use has raised concerns—some logical
DNA. An alternative technique is to introduce the DNA by and others not. For example, some people have expressed fear over
electroporation, a procedure that subjects the cells to an electric the fact that GM foods “contain DNA.” Considering that DNA is
current. DNA-mediated transformation, p. 202 consumed routinely as we eat plants and animals, this is obviously
After the DNA is introduced into the new host, the trans- an irrational concern. Others worry that unanticipated allergens
formed bacteria are grown on a medium that both selects for cells could be introduced into food products, posing a health threat. To
containing vector sequences and differentiates those carrying address this issue, the FDA has implemented strict guidelines, such
recombinant plasmids (figure  9.9). If the vector’s selectable as requiring producers to show that GM products intended for
marker encodes ampicillin resistance, for example, then the trans- human consumption do not unduly elicit allergic reactions.
formed cells are grown on ampicillin-containing medium. If the Incidents such as the inadvertent use of GM corn not approved for
second genetic marker is the lacZ gene, then X-gal is added to the human consumption in tortilla chips, however, continue to fuel
medium as well. Cells that form white colonies (rather than blue) apprehension about the effectiveness of regulatory control.
should harbor a recombinant molecule; these are then further char- Another concern about GM products is their possible unin-
acterized to determine if they carry the gene of interest. tended effects on the environment. Some laboratory studies have
shown that pollen from plants genetically modified to produce
MicroAssessment 9.3 Bt toxin can inadvertently kill monarch butterflies; other studies,
Introns can be removed from eukaryotic DNA to create cDNA. however, have refuted the evidence. In addition, there are indica-
Vectors typically have an origin of replication, a selectable marker, tions that herbicide-resistance genes can be transferred to weeds,
a multiple-cloning site, and a second genetic marker. decreasing the usefulness of the herbicide. As with any new tech-
7. Explain the role of reverse transcriptase in cloning. nology, the impact of GM organisms will need to be carefully
8. Explain the role of ampicillin and the lacZ gene in cloning. scrutinized to avoid negative consequences.
9. What would happen if a cell took up a molecule of MicroByte
circular insert DNA only? + The Genetic Information Nondiscrimination Act (GINA) protects
Americans against discrimination in healthcare coverage and
employment based on their genome.

9.4 ■ Concerns Regarding Genetic MicroAssessment 9.4

Engineering and Other Concerns about genetic engineering and genomics are varied and
DNA Technologies include ethical issues. Potentially adverse impacts of genetically
modified organisms on human health and the environment are also a
Learning Outcome concern.
7. Describe some of the concerns regarding DNA technologies. 10. Describe two concerns regarding information that can be gained
by analyzing a person’s DNA.
Any new technology should be scrutinized to ensure it is safe and 11. Describe two concerns regarding the use of genetically modified
effective. When recombinant DNA technologies first made gene organisms.
cloning possible over three decades ago, controversies swirled
about their use and possible abuse. Even the scientists who devel-
oped the technologies were concerned about potential dangers. In 9.5 ■ DNA Sequencing
response, the National Institutes of Health (NIH) formed the
Recombinant DNA Advisory Committee (RAC) to develop guide- Learning Outcomes
lines for conducting research involving recombinant DNA tech- 8. Describe two applications of DNA sequencing.
niques and gene cloning. Today, we are enjoying the fruits of 9. Describe the automated dideoxy chain termination method of DNA
many of those technologies, as evidenced by the list of commer- sequencing.
cially available products in table 9.3. It should be noted, however,
that although the technologies can be used to make life-saving DNA sequencing is the process of determining the nucleotide
products, there is no guarantee they will not be used for malicious sequence of a DNA molecule. The Human Genome Project, the
purposes. Today, the idea that new infectious agents are being cre- completed undertaking to sequence the human genome, resulted in
ated for the purpose of bioterrorism is a disturbing possibility. highly automated and efficient techniques. These allowed scien-
Recent advances in genomics have generated new cause for tists to more readily determine the genomic sequence of other
concern, primarily involving ethical issues regarding the appropri- organisms, including both prokaryotes and eukaryotes, fueling the
ateness and confidentiality of information gained by analyzing a rapidly growing field of genomics. The resulting explosion of data
person’s DNA. For example, will it be in an individual’s best inter- spawned a new field to analyze the information—bioinformatics.
est to be told of a genetic life-terminating disease? Could such genomics, p. 184 bioinformatics, p. 184
 Part I Life and Death of Microorganisms 225

By determining the DNA sequence of a genome, the amino fast and efficient process. Several newer sequencing methods have
acid sequence of the encoded proteins can be established. This been developed, and these are speeding the process even more.
makes it possible to compare characteristics of various proteins in The highly automated rapid methods are high throughput, mean-
different organisms. Non-coding sequences and mobile genetic ing that large numbers of samples can be processed very quickly.
elements can be compared as well. DNA sequencing also makes it
possible to determine the evolutionary relatedness of organisms, a Dideoxy Chain Termination Method
topic discussed in chapter 10. mobile genetic elements, p. 208 The fundamental aspect of the dideoxy chain termination method
Sequencing technologies have become so efficient that a new is an in vitro DNA synthesis reaction. It requires:
project has been initiated—the Human Microbiome Project—
■ Template DNA: Single-stranded DNA from which comple-
which uses genomics to determine the biological diversity in the
mentary copies are synthesized. template, p. 163
normal microbiota of the human body. This project will use both
traditional approaches (sequencing the genome of individual micro- ■ DNA polymerase: The enzyme that synthesizes DNA.
 DNA polymerase, p. 166
bial strains) and metagenomics. Goals of the Human Microbiome
Project include comparing the composition of the microbiota in ■ Primer: Short DNA molecules that anneal to the complemen-
health and disease, and determining the extent of its variation in tary sequence in the single-stranded template molecules. The
different individuals. normal microbiota, p. 8  metagenomics, p. 184 primer allows the researcher to choose where synthesis starts
(figure 9.10). Recall that DNA polymerase can add nucleo-
MicroByte tides only to an existing fragment of DNA, thereby extending
Sequencing the first human genome took 13 years and $440,000,000; the length of fragment. primer, p. 166
a race is on to sequence 100 human genomes in 10 days for about
■ Deoxynucleotides: Each of the four deoxynucleotides used in
$10,000 each.
DNA synthesis—dATP, dGTP, dCTP, and dTTP.
■ Dideoxynucleotides: These are identical to their deoxynucle-
Techniques Used in DNA Sequencing otide counterparts except they lack the 3OH group.
The most widely used technique for sequencing DNA is the If only the first four ingredients were in the reaction, full-length
dideoxy chain termination method. The procedure is now typi- molecules complementary to the template DNA would always
cally done using automated sequencing instruments, making it a be synthesized. The dideoxynucleotides, however, function as
chain terminators. They lack the chemical group to which sub-
sequent nucleotides would be added. Therefore, when a dideoxy-
nucleotide is added to a growing strand of DNA, no additional
Primer
nucleotides can be incorporated. Elongation of that strand stops
5' 3' (figure 9.11).
T G G C C G

T T G C C G T A C C G G C C C T T A A A T T G A A T
3' 5'
Primer is added to single-stranded DNA. PO4 OH Incorporation of a
deoxynucleotide (dNTP)
5' 3'OH elongates the chain. A
subsequent nucleotide can
3' 5' be added to the 3'OH.
DNA polymerase Nucleotide

5' 3' G A C
T G G C C G T PO4 H
T T G C C G T A C C G G C C C T T A A A T T G A A T Incorporation of a
3' 5' 5' 3'OH dideoxynucleotide (ddNTP);
the ddNTP lacks a 3'OH.
Primer anneals to complementary sequence, serving as the nucleic acid 3' 5'
fragment to which DNA polymerase can add nucleotides.

PO4 Chain elongation is

5' 3' H OH terminated. No additional
T G G C C G G G A A T T T A A C T T A 5' nucleotides can be added
3'
T T G C C G T A C C G G C C C T T A A A T T G A A T
5' 3'
X 5' due to the lack of a 3'OH.
Final product

FIGURE 9.11 Chain Termination by a Dideoxynucleotide

FIGURE 9.10 Primers Through appropriate primer selection, a Once a dideoxynucleotide is incorporated into a growing strand,
researcher can choose the site where in vitro DNA synthesis will start. additional nucleotides cannot be added.
? Why are primers required in DNA sequencing reactions? ? How is a dideoxynucleotide different from a deoxynucleotide?
226 Chapter 9 Biotechnology and Recombinant DNA

FIGURE 9.12 Dideoxy Chain

Termination Method of 1 Key ingredients in the reaction:
DNA Sequencing This figure • Primer and template (shown annealed)
illustrates the principles of the
automated method, which uses ATA G CTA C CTA G
dideoxynucleotides that carry a
fluorescent label. • DNA polymerase
• Deoxynucleotides (dATP, dTTP, dGTP, dCTP)
? What would happen if the • Fluorescently labeled dideoxynucleotides (ddATP, ddTTP, ddGTP, ddCTP; a very small amount of each)
deoxynucleotides were left
out of the reaction?
2 Chain elongation is terminated randomly 3 The fragments are denatured, and the single
when DNA polymerase incorporates a strands then separated by gel electrophoresis.
dideoxynucleotide (indicated by a colored The color of the fluorescent marker indicates
box and an asterisk). the terminating dideoxynucleotide.
Products of the reaction: Results:

T A T C G A T G GA T C*
ATA G CTA C CTAG

T A T C G A T GG A T*
ATA G CTA C CTA G

T A T C G A T G G A*
ATA G CTA C CTA G

T A T C G A T G G*
ATA G CTA C CTA G

T A T C G A T G*
ATA G CTA C CTA G

Decreasing fragment size

T A T C G A T*
ATA G CTA C CTA G

T A T C G A*
ATA G CTA C CTA G

T A T C G*
ATA G CTA C CTA G

T A T C*
ATA G CTA C CTA G

T A T*
ATA G CTA C CTA G

T A*
ATA G CTA C CTA G

T*
ATA G CTA C CTA G
 Part I Life and Death of Microorganisms 227

In an automated dideoxy sequencing reaction, DNA poly- 9.6 ■ Polymerase Chain

merase, template DNA, primer, the four deoxynucleotides, and a
very small amount of the four dideoxynucleotides are mixed Reaction (PCR)
together (figure 9.12 1 ). The different dideoxynucleotides each
carry a distinct fluorescent marker. When the reaction is incubated Learning Outcomes
at an appropriate temperature, the primers anneal to template 10. Describe how PCR can be used to diagnose diseases.
molecules and DNA synthesis begins. 2 Each nucleotide chain is 11. Explain how PCR can be used to exponentially amplify a select
elongated until a chain terminator—a dideoxynucleotide—is region of DNA.
incorporated. Termination happens infrequently, however, because
of the small amount of ddNTPs relative to dNTPs. The significant The polymerase chain reaction (PCR) makes it possible to cre-
aspect is that the color of the fluorescent marker carried by the ate more than a billion copies of a given region of DNA—referred
chain terminator can be used to determine which nucleotide was to as target DNA—in a matter of hours. In fact, target DNA can
incorporated at the terminating position. be generated in sufficient concentration to be visible to the
After the sequencing reaction, the sample is heated, which unaided eye when fragments in the sample are separated by gel
denatures the DNA. 3 Electrophoresis is then used to separate electrophoresis, stained with ethidium bromide, and illuminated
the DNA fragments and determine their relative size. The condi- with UV light (figure 9.14).
tions (pH, temperature, and gel concentration) keep the DNA One important application of PCR is disease diagnosis. For
single-stranded and allow fragments that differ in length by only example, if a woman is suspected of having the sexually transmit-
one nucleotide to be separated. A laser is used to detect the colors ted disease gonorrhea, PCR can be used to detect the causative
of fluorescent bands as they run past, recording their intensity as agent (Neisseria gonorrhoeae) in a vaginal specimen. This is done
a peak. The order of the colored peaks reflects the nucleotide by treating the specimen to release the DNA in cells and then
sequence of the DNA (figure 9.13). amplify target DNA unique to Neisseria gonorrhoeae. If that
DNA is amplified, then the pathogen must be present. PCR can
also be used to detect HIV nucleotide sequences in a sample of
MicroAssessment 9.5 blood cells, thereby diagnosing HIV infection.
Efficient DNA sequencing methods have fueled the rapidly growing
field of genomics. The automated dideoxy chain termination method
is a widely used sequencing technique.
12. What is the Human Microbiome Project?
13. How does a ddNTP terminate DNA synthesis?
DNA from Patient A DNA from Patient B
14. What would happen in the sequencing reaction if the relative
concentration of a dideoxynucleotide were increased? +
PCR amplifies
specific sequence
of interest.

No DNA amplified

Gel electrophoresis of
PCR amplified samples
Patient A Patient B

Conclusion:
Patient A is positive (infected);
FIGURE 9.13 Results of Automated DNA Sequencing The Patient B is negative.
order of the colored peaks reflects the nucleotide sequence of
the DNA. FIGURE 9.14 PCR Amplifies Selected Sequences
? What allows the peaks to be different colors? ? How can PCR be used to diagnose an infectious disease?
228 Chapter 9 Biotechnology and Recombinant DNA

Techniques Used in PCR

5' 3'
The polymerase chain reaction starts with a double-stranded Region of DNA to be amplified
3' 5'
DNA molecule that serves as a template from which more than
a billion identical copies of the target DNA can be produced.
The process involves DNA synthesis reactions and the follow-
ing key ingredients: 1 Heating to 95°C denatures DNA.

■ Double-stranded DNA: This contains the target DNA and

serves as a template for DNA synthesis. 5' 3'
■ Taq polymerase: This heat-stable DNA polymerase is from Primer Primer
the thermophile Thermus aquaticus.
3' 5'
■ Primers: As with the DNA sequencing reaction, the primers
allow the researcher to choose where synthesis starts.
■ Deoxynucleotides: Each of the four deoxynucleotides used in 2 Cooling to 50°C allows the added
DNA synthesis—dATP, dGTP, dCTP, and dTTP. primers to anneal to the single-
stranded templates.

The Three-Step Amplification Cycle

5' 3'
PCR uses a repeating cycle consisting of three steps (figure 9.15).
3' 5'
1 In the first step, the sample is heated to near-boiling (about
5' 3'
95°C) in order to denature the DNA. 2 In the second step, the 3' 5'
temperature is lowered (to about 50°C); within seconds, the prim-
ers anneal to their complementary sequences on the denatured
target DNA. 3 In the third step, the temperature is raised to the
3 DNA synthesis occurs when the
optimal temperature of Taq DNA polymerase (about 70°C), allow- temperature is raised to 72°C.
ing DNA synthesis to occur. After one three-step cycle, the target
DNA is duplicated.
In subsequent amplification cycles, the original DNA strands
5' 3'
serve as templates again, and so do the newly synthesized strands.
3' 5'
Because the number of template molecules increases with each
5' 3'
cycle, PCR amplifies the target exponentially. After a single cycle 3' 5'
of the three-step reaction, there will be two double-stranded DNA
molecules for every original; after the next cycle, there will be
four; after the next there will be eight, and so on.
A critical factor in PCR is Taq polymerase, the heat-stable
DNA polymerase of Thermus aquaticus. This polymerase, unlike
5' 3'
the DNA polymerase of E. coli, is not destroyed at the high 3' 5'
temperature used to denature the DNA in the first step of each
amplification cycle. If a heat-stable polymerase were not used, 5' 3'
3' 5'
fresh polymerase would need to be added after that step in each
cycle of the reaction. The discovery and characterization of The products of one 3-step cycle of PCR
T. aquaticus through basic research was key to developing this
widely used and commercially valuable method. thermophile,
p. 90 FIGURE 9.15 Steps of a Single Cycle of PCR
? Considering that over a billion copies of target DNA can be made
using PCR in only a matter of hours, approximately how many
molecules of primers must be included in the reaction?
Generating the PCR Product
Although the preceding description explains how PCR ampli-
fies the target DNA exponentially, it does not clarify how a
fragment containing only the target DNA becomes the pre-
dominant product. This fragment is referred to as the PCR amplified target (PCR product) will appear as a single band on
product. Generating the PCR product is important, because it the ethidium bromide–treated gel.
allows the technician to use PCR coupled with gel electropho- To understand how the PCR product is generated, you must
resis to detect the target DNA in a sample. After PCR, the consider the exact sites to which the primers anneal, and visualize
 Part I Life and Death of Microorganisms 229

PERSPECTIVE 9.1
Science Takes the Witness Stand
After serving more than 10 years on a rape samples and is quite time-consuming. Most makes tandem repeats a useful genetic marker
charge, a wrongfully convicted young man forensics labs have now switched to using a for distinguishing individuals. With PCR,
was released from prison when a new DNA PCR-based method because results can be using primers that bind regions flanking the
typing technique exonerated him. The new obtained in less than 5 hours from a sample as repeating sequences, the number of repeats can
technique, based on using the polymerase small as a drop of blood the size of a pinhead. be determined. A fragment that contains 9
chain reaction (PCR) to amplify specific In fact, the FBI now catalogs PCR-based DNA repeats, for example, will be longer than one
sequences, showed that the semen sample profiles from unsolved crimes and convicted that contains only 7. The PCR-amplified frag-
taken from the rape victim did not contain the violent offenders, making it easier to track or ments can be quickly separated using a rapid
man’s DNA. Indeed, a database indicated a link the crimes of serial offenders. The national type of gel electrophoresis called capillary
DNA match with a man currently in prison for database is called CODIS (Combined DNA electrophoresis. A size standard is incorpo-
an unrelated rape charge. Index System). rated so that the sizes of the PCR-amplified
Stories abound about the growing power PCR-based DNA typing amplifies certain fragments can be determined.
of DNA evidence for obtaining convictions chromosomal regions that contain short tandem The FBI’s CODIS database catalogs the
and also clearing the wrongly accused, but how repeats (STRs). These consist of a core sequence amplification pattern of 13 different STR loci
is DNA used in forensics? It is not practical to of two to six base pairs that repeat a variable (chromosomal locations). Commercially avail-
compare the entire nucleotide sequence of two number of times in different people. On chro- able kits contain fluorescently labeled primers
people; instead, specific regions that vary sig- mosome 2, for example, in an intron within the that allow simultaneous amplification and
nificantly between individuals are analyzed. In thyroid peroxidase gene, the sequence AATG is subsequent recognition of each of the 13 loci.
the past, forensics labs have used a method that repeated sequentially between 5 to 14 times. In A laser detects the color of each amplified
employs the probe-based technique called one individual, there may be 9 of these STRs in fragment as it moves out of the capillary gel,
Southern blot hybridization to detect differ- one copy of that chromosome and 7 in the other, and computer analysis generates a pattern of
ences. This method provides valuable informa- whereas another individual may have 11 and 5 peaks that reflect the STR profile of the DNA
tion but cannot be used on small or degraded (figure  1). This variation, or polymorphism, sample.

Individual A
Chromosomal
copy 1 A ATG
(9 copies of T TAC
the STR)

Amplified regions
Tandem repeats
Sequences used as PCR primers
Chromosomal copy 2
(7 copies of the STR)

Individual B
Chromosomal
copy 1 A ATG
(11 copies of T TAC
the STR)

Chromosomal copy 2
(5 copies of the STR)

FIGURE 1 Using PCR to Type DNA PCR is used to amplify certain chromosomal regions containing short tandem repeats (STRs). The
number of copies of a given STR varies among people, resulting in corresponding differences in the length of the amplified fragments.
Typically, at least 13 different STR locations are analyzed.
230 Chapter 9 Biotechnology and Recombinant DNA

at least three cycles of PCR (figure 9.16). 1 In the first cycle, 5 and 3 ends of this strand are determined by the sites to which
two new strands are generated. Note, however, that their 5 ends the primers initially annealed.
are primer DNA; the strands are shorter than the original templates In the third round of replication, the full-length and the
but longer than the target DNA. These mid-length strands will be mid-length strands again will be used as templates, repeating
generated whenever the original full-length molecule is used as a the processes just described. 3 The short strands generated in
template, which is once per three-step cycle. the preceding round will also be used as templates, however,
In the next cycle, the full-length molecules will again be generating short double-stranded molecules—the PCR prod-
used as templates, repeating the process just described. 2 More uct. Continuing to follow the events in further rounds of repli-
importantly, the mid-length strands created during the first cycle cation will reveal that this fragment is exponentially amplified
will be used as templates for DNA synthesis. As before, the (figure 9.17).
primers will anneal to these strands and then nucleotides will be
added to the 3 end. Elongation, however, will stop at the 5 end
of the template molecule, because DNA synthesis requires a Selecting Primer Pairs
template. Recall that the 5 end of the template is primer DNA. The nucleotide sequences of the two primers are critical because
Thus, whenever a mid-length strand is used as a template, a short the primers dictate which portion of the DNA is amplified. Each
strand—exactly the length of the target DNA—is generated. The must be complementary to one end of the target DNA, so that

1 Target DNA
Mid-length strands are synthesized
Template from the full-length templates. The
5' sites to which the primers annealed
3'
determine the 5' ends of these strands.
3' 5'
Mid-length Primer
Outcome of cycle 1 fragment
(as well as subsequent cycles) Mid-length
Primer fragment
5' 3'
3' 5'
Template

2 Short fragment When the mid-length strands made

5' 3' in the reaction above are used as
3' 5' templates, short strands are
Cycle 1 mid-length generated. The 3' ends of the
fragment (template) fragments are complementary to the
Outcome of cycle 2 Cycle 1 mid-length 5' ends of the primers.
(as well as subsequent cycles) fragment (template)
5' 3'
3' 5'
Short fragment

3 Short fragment When short strands made in the

5' 3' reaction above are used as
3' 5' templates, like-sized strands are
Cycle 2 short generated. This is the double-
fragment (template) stranded target molecule that will be
Outcome of cycle 3 Cycle 2 short amplified exponentially.
(as well as subsequent cycles) fragment (template)
5' 3' Produced when full-length molecule is used as a template
3' 5' Produced when mid-length molecule is used as a template
Short fragment Produced when target fragment is used as a template

FIGURE 9.16 The PCR Product Is a Fragment of Discrete Size The positions to which the primers anneal to the template dictate the
size and sequence of the fragment amplified exponentially.
? If two mid-length strands are present at the end of cycle 1, how many will be present at the end of cycle 3?
 Part I Life and Death of Microorganisms 231

Time 0 End of cycle 1 End of cycle 2 End of cycle 3 End of cycle 4 End of cycle 5

(2)

(8)

(22)

FIGURE 9.17 Exponential Amplification of Target DNA During PCR, mid-length fragments are amplified linearly (arithmetically),
whereas the discrete-sized target DNA, referred to as PCR product, is amplified exponentially. After 30 cycles of PCR, more than a billion molecules
of PCR product will have been synthesized.
? How many PCR cycles are required to generate double-stranded target fragments?
232 Chapter 9 Biotechnology and Recombinant DNA

DNA synthesis will extend across that stretch of DNA. If a techni-

Probe
cian wants to amplify a DNA sequence that encodes a specific Label
protein, he or she must first determine the nucleotide sequences at 5' 3'
the ends of the gene. That information can be used to obtain the T G G C C G

appropriate pair of primers. T T G C C G T A C C G G C C C T T A A A T T G A A T

3' 5'

MicroAssessment 9.6 Probe is added to single-stranded DNA that has been attached to a
solid surface.
The polymerase chain reaction (PCR) is used to rapidly increase the
amount of a specific segment of DNA in a sample.
15. What is the role of Thermus aquaticus in PCR?
16. Explain why it is important to use a polymerase from a
thermophile in the PCR reaction.
5' 3'
17. Sequencing reactions can be done using PCR. In this case, T G G C C G
would two primers be necessary? Explain. + T T G C C G T A C C G G C C C T T A A A T T G A A T
3' 5'
Probe anneals to complementary sequence. Because of the label it
carries, its location can easily be determined.

9.7 ■ Probe Technologies FIGURE 9.18 DNA Probes These single-stranded pieces of
DNA tagged (labeled) with a detectable marker are used to detect
specific nucleotide sequences in DNA or RNA samples that have been
Learning Outcome
attached to a solid surface.
12. Compare and contrast the applications and techniques of colony
blotting, FISH, and DNA microarray technologies. ? Why is it important that the probe be labeled?

DNA probes are used to locate specific nucleotide sequences in

nucleic acid samples attached to a solid surface. The probe is a
single-stranded piece of DNA, complementary to the sequence of membrane and incubated under conditions that allow the probe
interest, that has been labeled with a detectable marker such as a to hybridize to complementary sequences on the filter. Any
radioactive isotope or a fluorescent dye. The probe will anneal to probe that has not bound is then washed off. 5 The appropriate
its complement, a process called hybridization. By hybridizing to method is then used to detect the label carried by the probe,
its complement, the probe “finds” the sequence of interest and thereby locating the position of the hybridized probe. The posi-
makes it detectable (figure 9.18). tions to which the probe hybridized indicate colonies that have
A variety of technologies use DNA probes to locate specific the DNA of interest.
nucleotide sequences. They include colony blotting, fluores-
cence in situ hybridization (FISH), and DNA microarrays. The
technique called Southern blotting also uses probes, but its appli- Fluorescence in situ
cations have been largely replaced by PCR. For a description of
Hybridization (FISH)
Southern blotting, visit the text website (www.mhhe.com/
nester7). Fluorescence in situ hybridization (FISH) uses a fluorescently
labeled probe to detect specific nucleotide sequences within intact
cells attached to a microscope slide. Cells containing the hybrid-
ized probe can then be observed using a fluorescence microscope.
Colony Blotting To study prokaryotes, a probe that hybridizes to sequences on
Colony blotting uses probes to detect specific DNA sequences ribosomal RNA (rRNA) is generally used. This is because multi-
in colonies grown on agar plates (figure 9.19 1 ). This method plying cells can have thousands of copies of rRNA, increasing the
is commonly used to determine which clones in a DNA library technique’s sensitivity. Other characteristics of rRNA that make it
or other collection contain a sequence being studied. 2 The useful for identifying prokaryotes are described in chapter 10.
term “blot” in the name reflects the fact that the colonies are  fluorescence microscope, p. 44
transferred in place (“blotted”) onto a nylon membrane, creating FISH is revolutionizing microbial ecology research and holds
a replica of the colonies on the original plate. The membrane great promise in clinical laboratories. It provides a means to rap-
serves as a durable, permanent support for the cells of the colo- idly identify microorganisms directly in a specimen, bypassing the
nies and their DNA. 3 After the transfer, the membrane is need to grow them in culture. FISH can be used to detect either a
soaked in an alkaline solution to simultaneously lyse the cells group of related organisms or a specific species, depending on the
and denature their DNA, generating single-stranded DNA mol- nucleotide sequence of the probe. For example, FISH can be used
ecules. 4 A solution containing the probe is then added to the to determine the relative proportion of two different groups of
 Part I Life and Death of Microorganisms 233

Nylon membrane
Probe bound to DNA

1 Colonies on an agar 2 Colonies are transferred 3 The membrane is 4 Probe is added that 5 By locating the
plate. in place (“blotted”) to a soaked in an alkaline binds to DNA of positions to which
nylon membrane. solution to lyse the cells interest. probe has bound,
and denature their colonies that have the
DNA. DNA of interest can be
located.

FIGURE 9.19 Colony Blotting This technique is used to determine which colonies on an agar plate contain a given DNA sequence.
? What is the purpose of soaking the membrane in an alkaline solution?

prokaryotes in the same specimen by using two separate probes— To analyze a sample using FISH, the sample must first be
one specific for each group—labeled with different colored fluo- treated with chemicals to preserve the shape of the cells, inactivate
rescent markers (figure 9.20). It can also be used to identify cells enzymes that might otherwise degrade the nucleic acid, and make
of the bacterium that causes tuberculosis in a sputum specimen. the cells more permeable so that the labeled probe molecules can
 microbial ecology, p. 719 tuberculosis, p. 502 easily enter. Once the specimen has been prepared, it is put on a
glass slide, bathed with a solution containing the labeled probe,
and incubated under conditions that allow hybridization to occur.
Unbound probe is then washed off. Finally, the specimen is
viewed using a fluorescence microscope.

DNA Microarrays
DNA microarrays are primarily used to study gene expres-
sion in organisms whose genomes have been sequenced.
They can also be used to detect a specific nucleotide
sequence in a DNA sample of interest.
A DNA microarray is a glass slide or other small solid
support carrying an arrangement of tens or hundreds of thou-
sands of short DNA fragments. Each DNA fragment functions
in a manner analogous to a probe, allowing a researcher to screen
a single sample for a vast range of different sequences
simultaneously. Unlike typical probes, however, the
arrays do not carry a detectable label. Instead, the label
is on the nucleic acid of interest.
To study gene expression, mRNA is isolated from
an organism and converted to fluorescently labeled,
single-stranded cDNA. Those molecules are then allowed
to hybridize to a microarray specially created to include sequences
FIGURE 9.20 Fluorescence in situ Hybridization (FISH) Two specific for each gene of a particular organism. The locations of
different probes have been used to stain the cells. The probe the labeled cDNA molecules can be detected using a computer-
that hybridizes to Ignicoccus rRNA fluoresces green; the one that ized scanner. The genes to which cDNAs hybridize are ones that
hybridizes to Nanoarchaeum rRNA fluoresces red. Size bar = 1 μm. the organism was expressing. By doing the experiment using cul-
? Why does FISH typically require a probe that hybridizes to rRNA? tures grown under different sets of conditions, and labeling their
234 Chapter 9 Biotechnology and Recombinant DNA

cDNAs with different fluorescent markers, variations in gene

expression can be revealed (figure 9.21). cDNA, p. 222
To use microarrays to detect specific nucleotide sequences in
an organism whose genome has not yet been sequenced, the DNA
is digested into small fragments, labeled with a fluorescent
marker, denatured, and then added to an appropriate microarray.
Because the sequence of each of the fragments that make up the
array is known, the location of the label can be used to determine
the presence of specific sequences in the DNA of interest.

MicroByte
Using DNA microarrays, researchers discovered that pathogens
express some genes only when in certain locations within the human
body.

MicroAssessment 9.7
Colony blotting uses probes to identify colonies that contain a
given sequence of DNA. Fluorescence in situ hybridization is FIGURE 9.21 A DNA Microarray This is an example of an
used to observe individual cells that contain a given sequence. array used to study gene expression. Two different cDNA samples
DNA microarrays allow researchers to study gene expression (one labeled with a red fluorescent marker and the other with
a green fluorescent marker) were simultaneously hybridized to
and to screen a sample for a vast range of different sequences
fragments in the microarray. Red dots indicate the positions to which
simultaneously.
one sample hybridized, and green dots indicate the positions to
18. What role does colony blotting play in cloning? which the other hybridized. Yellow dots indicate that both samples
19. Why is a probe that binds to rRNA used in FISH? hybridized.

20. In FISH, what would happen if unbound probe was not ? How can DNA microarrays be used to determine which genes of
washed off? + a bacterial pathogen are expressed both inside and outside of a
host cell?

Summary
9.1 ■ Fundamental Tools Used in Biotechnology Obtaining DNA

Restriction Enzymes (figure 9.1) To isolate DNA, cells are lysed by adding a detergent. To obtain
eukaryotic DNA without introns, reverse transcriptase is used to make
Restriction enzymes cut DNA into fragments. Cohesive ends will
a cDNA from an mRNA template (figure 9.7).
anneal to one another, making it possible to join DNA from two dif-
ferent organisms. Generating a Recombinant DNA Molecule

Gel Electrophoresis (figure 9.2) DNA ligase is used to join the vector and the insert (figures 9.8, 9.9).
Gel electrophoresis separates DNA fragments according to their size. Obtaining Cells That Harbor Recombinant DNA Molecules
The recombinant molecule is introduced into the new host, usually
9.2 ■ Applications of Genetic Engineering (table 9.3) E. coli, using transformation or electroporation. The transformed cells
Genetically Engineered Bacteria (figures 9.3, 9.4) are grown on medium that both selects for cells containing vector
sequences and differentiates those that carry recombinant molecules.
Bacteria can be engineered to produce pharmaceutical proteins, vac-
cines, and other proteins more efficiently. By cloning a segment of ■
9.4 Concerns Regarding Genetic Engineering
DNA into E. coli, an easy source of that sequence is available for study
and Other DNA Technologies
and further manipulation.
Advances in genomics raise ethical issues and concerns about confiden-
Genetically Engineered Eukaryotes tiality. Genetically modified organisms hold many promises, but con-
Transgenic plants have been engineered to resist pests and herbicides, cerns exist about the inadvertent introduction of allergens into a food
have improved nutritional value, and function as edible vaccines. product and adverse effects on the environment.

9.3 ■ Techniques Used in Genetic Engineering 9.5 ■ DNA Sequencing

A DNA library is a collection of clones that together contain the entire By sequencing DNA, the encoded information can be compared to that
genome of an organism (figure 9.6). of other organisms.
 Part I Life and Death of Microorganisms 235

Techniques Used in DNA Sequencing 9.7 ■ Probe Technologies

A key ingredient in the sequencing reaction is a dideoxynucleotide, a DNA probes are used to locate specific nucleotide sequences
nucleotide that lacks the 3OH and therefore functions as a chain ter- (figure 9.18).
minator (figure  9.11). The sizes of fragments in a sequencing reaction
indicate the positions of the terminating nucleotide (figures 9.12, 9.13). Colony Blotting
Colony blotting uses a probe to identify colonies that contain a given
sequence of DNA (figure 9.19).
9.6 ■ Polymerase Chain Reaction (PCR)
PCR is used to rapidly increase the amount of a specific DNA segment Fluorescence in situ Hybridization (FISH)
in a sample (figure 9.14). Fluorescence in situ hybridization (FISH) uses a fluorescently labeled
probe to detect specific nucleotide sequences within intact cells affixed
Techniques Used in PCR to a microscope slide (figure 9.20).
Double-stranded DNA is denatured, primers anneal to their comple-
mentary sequences, and then DNA is synthesized, amplifying the target DNA Microarrays
sequence (figure  9.15). The PCR product is amplified exponentially DNA microarrays contain tens or hundreds of thousands of
(figures 9.16, 9.17). The primers dictate which portion of the DNA is oligonucleotides that each function in a manner analogous to a probe
amplified. (figure 9.21).

Review Questions
Short Answer d) a multiple-cloning site.
1. Why are restriction enzymes useful in biotechnology? e) the lacZ gene.
2. Describe three general uses of genetically engineered bacteria. 5. Which of the following describes the function of the lacZ gene in
a cloning vector?
3. Describe the function of a reporter gene.
a) Means of selecting for cells that contain vector sequences
4. Describe three uses of genetically engineered plants.
b) Means of distinguishing cells that have taken up recombinant
5. What is a DNA library? molecules
6. What is cDNA? Why is it used when cloning eukaryotic genes? c) Site required for the vector to replicate
7. How many different temperatures are used in each cycle of the d) Mechanism by which cells take up the DNA
polymerase chain reaction? e) Gene for a critical nutrient required by transformed cells
8. Explain how PCR eventually generates a discrete-sized fragment 6. Which is used for cloning eukaryotic genes but not prokaryotic
from a much longer piece of DNA. genes?
9. Describe the function of a probe. a) Restriction enzymes
b) DNA ligase
10. How does a DNA microarray function as a set of probes?
c) Reverse transcriptase
d) Vector
Multiple Choice e) Selectable marker
7. Which of the following does a dideoxynucleotide lack?
1. What is the function of a vector?
a) 5PO4 b) 3OH c) 5OH
a) Destroys cells that do not contain cloned DNA
d) 3PO4 e) c and d
b) Allows cells to take up foreign DNA
8. In a sequencing reaction, the dATP was left out of the tube. What
c) Carries cloned DNA, allowing it to replicate in cells
would be the result of this error?
d) Encodes herbicide resistance
a) No synthesis would occur.
e) Encodes Bt toxin
b) Synthesis would never continue past the first A.
2. The Ti plasmid of Agrobacterium tumefaciens is used to geneti-
c) Synthesis would not stop until the end of the template.
cally engineer which of the following cell types?
d) Synthesis would terminate randomly, regardless of the
a) Animals b) Bacteria c) Plants
nucleotide incorporated.
d) Yeast e) All of these e) The error would have no effect.
3. Which of the following can be used to generate a DNA library? 9. The polymerase chain reaction uses Taq polymerase rather than a
a) PCR b) Sequencing c) Colony blotting DNA polymerase from E. coli, because Taq polymerase
d) Microarrays e) Cloning a) introduces fewer errors during DNA synthesis.
4. An ideal vector has all of the following except b) is heat-stable.
a) an origin of replication. c) can initiate DNA synthesis at a wider variety of sequences.
b) a gene encoding a restriction enzyme. d) can denature a double-stranded DNA template.
c) a gene encoding resistance to an antibiotic. e) is easier to obtain.
236 Chapter 9 Biotechnology and Recombinant DNA

10. The polymerase chain reaction generates a fragment of a distinct civilization. What was the rationale for the argument by the
size even when an intact chromosome is used as a template. What second student?
determines the boundaries of the amplified fragment? 2. A student wants to clone gene X. On both sides of the gene are
a) The concentration of one particular deoxynucleotide in the the recognition sequences for AluI and BamHI (look at table  9.2).
reaction Which enzyme would be easier to use for the cloning experiment
b) The duration of the elongation step in each cycle and why?
c) The position of a termination sequence, which causes the Taq
polymerase to fall off the template
d) The sites to which the primers anneal Critical Thinking +
e) The temperature of the elongation step in each cycle 1. Discuss some potential issues regarding gene therapy, the use of
genetic engineering to correct genetic defects.
Applications 2. An effective DNA probe can sometimes be developed by knowing
1. Two students in a microbiology class are arguing about the ori- the amino acid sequence of the protein encoded by the gene. A
gins of biotechnology. One student argued that biotechnology student argued that this is too time-consuming since the complete
started with the advent of genetic engineering. The other stu- amino acid sequence must be determined in order to create the
dent disagreed, saying that biotechnology was as old as ancient probe. Does the student have a valid argument? Why or why not?
10
Identification and Classification
of Prokaryotic Organisms
KEY TERMS
Classification The process of Nomenclature The system of
arranging organisms into similar or assigning names to organisms.
related groups (taxa), primarily to Phylogeny Evolutionary
provide easy identification and study. relatedness of organisms.
Dichotomous Key Flowchart of Signature Sequence
tests used to identify organisms. Characteristic sequences in the
Domain A collection of similar ribosomal RNA genes, or their
kingdoms; there are three domains— products, that can be used to classify
Bacteria, Archaea, and Eucarya. or identify certain organisms.
Genus A collection of related Species A group of closely related
species. isolates or strains; the basic unit of
taxonomy.
Horizontal Gene Transfer
Transfer of DNA from one organism Strain An isolate; subgroup within
to another through conjugation, a species.
DNA-mediated transformation, or Taxonomy The science that studies
transduction. organisms in order to arrange them
Identification The process of into groups (taxa); involves three
characterizing an isolate in order to interrelated areas—identification,
determine the group (taxon) to which classification, and nomenclature.
it belongs.

PulseNet tracks bacterial strains causing foodborne diseases.

In the late 1970s, Carl Woese and his colleagues at the University of
A Glimpse of History Illinois determined the nucleotide sequence of ribosomal RNA in a wide
In the early 1870s, the German botanist Ferdinand Cohn published several variety of organisms. Based on the data, they recognized that prokaryotes
papers on bacterial classification, grouping microorganisms according to could be divided into two major groups that differ from one another as
shape—spherical, short rods, elongated rods, and spirals. That scheme much as they do eukaryotic cells. This led to a revolutionary system of
was not adequate, however, because there were too many different kinds classification that separates prokaryotes into two domains—the Archaea
of bacteria that had similar shapes. and the Bacteria. Each of these is on the same level as the Eucarya, which
The second major attempt at bacterial classification was initiated by includes the animals, plants, and fungi (all eukaryotes).
Sigurd Orla-Jensen. His early training in Copenhagen was in chemical
engineering, but he soon became interested in microbiology. In 1908,

I 
he proposed that bacteria be classified according to their physiological nformation that is logically organized is easier to use. News-
properties rather than morphology. papers, for instance, do not scatter various subjects throughout
A quarter of a century later, two Dutch microbiologists, Albert Kluyver the paper; instead, the information is grouped by general topic
and C. B. van Niel, proposed classification systems based on evolutionary such as local news, sports, and entertainment. A large library
relationships. They recognized a very serious problem, however: There was would be extremely difficult to use if the locations of the many
no way to distinguish between “resemblance” and “relatedness.” The fact books were not organized by subject matter. Likewise, scientists
that two prokaryotes look alike does not mean they are genetically related. have sorted living organisms into different groups, the better to
In 1970, Roger Stanier, a microbiologist at the University of understand the relationships among the species.
California–Berkeley, pointed out that relationships could be determined
Take a moment and think about how you would group bac-
by comparing either physical traits, such as proteins and cell walls, or
teria if you were to create a classification system. Would you
nucleotide sequences. At that time, most microbiologists, including Stanier,
assumed that all prokaryotes are basically similar. Chemical analysis of group them according to shape? Or would it make more sense to
structures from a wide variety of prokaryotes, however, showed that many consider their motility? Perhaps you would group them accord-
prokaryotes were fundamentally different from Escherichia coli, consid- ing to their medical significance. But then, how would you
ered a “typical” bacterium. These “unusual” features included the chemical classify two apparently identical organisms that differ in their
nature of the cell wall, cytoplasmic membrane, and ribosomal RNA. pathogenicity?

237
238 Chapter 10 Identification and Classification of Prokaryotic Organisms

10.1 ■ Principles of Taxonomy such as plants and animals, the basic taxonomic unit, a species,
is generally considered to be a group of morphologically similar
Learning Outcome organisms capable of interbreeding to produce fertile offspring.
1. Describe how prokaryotes are identified, classified, and assigned
Obviously, it is not possible to apply these same criteria to pro-
names. karyotes, which makes classification problematic.
Historically, taxonomists have relied heavily on phenotypic
Taxonomy is the science that studies organisms in order to arrange characteristics to classify prokaryotes. The development and appli-
them into groups (taxa). Those organisms with similar properties cation of molecular techniques such as nucleotide sequencing,
are grouped together and separated from ones that are different. however, is finally making it possible to determine the phylogeny
Taxonomy can be viewed as three separate but interrelated areas: of microorganisms. phenotype, p. 189

■ Identification: The process of characterizing an isolate Taxonomic Hierarchies

to determine the group (taxon) to which it belongs. Taxonomic classification categories are arranged in a hierarchical
■ Classification: The process of arranging organisms into simi- order, with the species being the basic unit. The species designa-
lar or related groups, primarily to make it easier to identify tion gives a formal taxonomic status to a group of related isolates,
and study them. or strains, which, in turn, permits their identification. Without
■ Nomenclature: The system of assigning names to organisms. classification, scientists and others would not be able to commu-
nicate about organisms with any degree of accuracy. Taxonomic
categories include:
Strategies Used to Identify Prokaryotes
■ Species: A group of closely related isolates or strains. Note
In practical terms, identifying the genus and species of a prokary- that members of a species are not all identical; individual
ote may be more important than understanding its genetic rela- strains may vary. The difficulty for the taxonomist is to decide
tionship to other microbes. For example, a food manufacturer is how different two isolates must be in order to be classified as
most interested in detecting microbial contaminants that can spoil separate species rather than strains of the same species.
a food product. In a clinical laboratory, identifying a pathogen
■ Genus: A collection of similar species.
quickly is critical so that a patient can be treated appropriately.
To identify microorganisms, a wide assortment of procedures ■ Family: A collection of similar genera. In prokaryotic nomen-
may be used, including microscopic examination, culture charac- clature, the name of the family ends in the suffix -aceae.
teristics, biochemical tests, and nucleic acid analysis. In a clinical ■ Order: A collection of similar families. In prokaryotic
laboratory, the patient’s disease symptoms play an important role nomenclature, the name of the order ends in the suffix -ales.
as well. For example, pneumonia in an otherwise healthy adult is ■ Class: A collection of similar orders.
typically caused by Streptococcus pneumoniae, a bacterium easily
■ Phylum or Division: A collection of similar classes.
differentiated from others using a few specific tests. In contrast,
diagnosing the cause of a wound infection is often more difficult, ■ Kingdom: A collection of similar phyla or divisions.
because several different microorganisms could be involved. ■ Domain: A collection of similar kingdoms. The domain is a
Often, however, it is only necessary to rule out the presence relatively new taxonomic category that reflects the character-
of organisms known to cause a particular disease, rather than istics of the cells that make up the organism.
conclusively identify each and every one. For instance, a fecal
specimen from a patient complaining of diarrhea and fever would Note, however, that microbiologists often group prokaryotes
generally be tested only for the presence of organisms that cause into informal categories based on one or more distinctive char-
those symptoms. The methods used to identify prokaryotes will acteristics, rather than using the higher taxonomic ranks such as
be discussed later in the chapter. Streptococcus pneumoniae, p. 497

Strategies Used to Classify Prokaryotes Taxonomic Ranks of the

TABLE 10.1
Understanding the evolutionary relatedness, or phylogeny, of Bacterium Escherichia coli
prokaryotes is important in creating a classification scheme that Formal Rank Example
reflects their evolution and biology. Such a scheme is more useful
than one that simply groups organisms by arbitrary characteristics, Domain Bacteria
because it is less prone to the bias of human perceptions. It also Phylum Proteobacteria
makes it easier to classify newly recognized species and allows Class Gammaproteobacteria
scientists to make predictions, such as which genes are likely to
Order Enterobacteriales
be transferred between organisms.
Family Enterobacteriaceae
Unfortunately, determining evolutionary relatedness among
prokaryotes is more difficult than it is for plants and animals. Genus Escherichia
Not only do prokaryotes have few differences in size and shape, Species coli
they do not undergo sexual reproduction. In higher organisms
 Part II The Microbial World 239

Bacteria Archaea Eucarya

Animals
Filamentous anoxygenic Entamoebae Slime
phototrophic bacteria molds
Fungi
Gram- Methanosarcina
Spirochetes positive
Methano- Halobacteria Plants
bacteria bacterium
Proteobacteria Thermococcus Methano-
Thermoproteus coccus Thermoplasma
Cyanobacteria Pyrodictium
Trypanosoma
Flavobacteria

Trichomonads

Thermotoga

Aquifex Microsporidia
Diplomonads

FIGURE 10.1 The Three-Domain System of Classification This classification system separates prokaryotic organisms into two domains—
Bacteria and Archaea. The third domain, Eucarya, contains all organisms composed of eukaryotic cells.
? Information about which molecule was used to establish the three-domain system of classification?

order, class, and phylum. Examples of such informal groupings the three domains—Bacteria, Archaea, and Eucarya (figure 10.1).
include the lactic acid bacteria, the anoxygenic phototrophs, the The system is based on the work of Carl Woese and colleagues,
endospore-formers, and the sulfate reducers. Organisms within who compared the nucleotide sequences in ribosomal RNA from
these groupings share similar phenotypic and physiological char- a wide variety of organisms (see A Glimpse of History). The
acteristics, but may not be genetically related. lactic acid bacteria, ribosomal RNA data are consistent with other differences between
p. 258 anoxygenic phototrophs, p. 259 sulfate reducers, p. 258 the Archaea and Bacteria, including the chemical compositions
An example of how a particular bacterial species is classified of their cell walls and cytoplasmic membranes (table  10.2).
is shown in table  10.1. The table intentionally omits the taxo- Bacteria, p. 9 Archaea, p. 9 Eucarya, p. 10
nomic category of kingdom, because the use of kingdoms within Before the three-domain system was introduced, the most
the Bacteria and Archaea is still in a state of flux. widely accepted scheme was the five-kingdom system, proposed
by R. H. Whittaker in 1969. The five kingdoms in this system are
Classification Systems Plantae, Animalia, Fungi, Protista (mostly single-celled eukary-
Classification systems change over the years as new information otes), and Prokaryotae. Although the five-kingdom system rec-
is discovered. There is no “official” classification system, and, as ognizes the obvious morphological differences between plants
new ones are introduced, others become outdated. The classification and animals, it does not reflect the recent genetic insights of the
scheme currently favored by most microbiologists is the three- ribosomal RNA data, which indicates that plants and animals are
domain system. This designates all organisms as belonging to one of more closely related to each other than Archaea are to Bacteria.

TABLE 10.2 A Comparison of Typical Properties of the Three Domains—Archaea, Bacteria, and Eucarya
Cell Feature Archaea Bacteria Eucarya

Peptidoglycan cell wall No Yes No

Cytoplasmic membrane lipids Hydrocarbons (not fatty acids) Fatty acids linked to glycerol Fatty acids linked to glycerol by
linked to glycerol by ether linkage by ester linkage ester linkage
Ribosomes 70S 70S 80S
Presence of introns Sometimes No Yes
Membrane-bound nucleus No No Yes
240 Chapter 10 Identification and Classification of Prokaryotic Organisms

TABLE 10.3 Taxonomic Outline of Bergey’s Manual of Systematic Bacteriology, 2nd edition
Representative Genera

Volume 1: The Archaea and the Deeply Branching Phototrophic Bacteria

Domain Archaea
Phylum Crenarchaeota Pyrodictium, Ignicoccus
Phylum Euryarchaeota Halobacterium, Methanococcus, Natronococcus, Picrophilus
Domain Bacteria
Phylum Aquificae Aquifex
Phylum Thermotogae Thermotoga
Phylum Thermodesulfobacteria Thermodesulfobacterium
Phylum Deinococcus–Thermus Deinococcus, Thermus
Phylum Chrysiogenetes Chrysiogenes
Phylum Chloroflexi Chloroflexus
Phylum Thermomicrobia Thermomicrobium
Phylum Nitrospira Nitrospira
Phylum Deferribacteres Deferribacter
Phylum Cyanobacteria Anabaena, Spirulina, Synechococcus
Phylum Chlorobi Chlorobium, Pelodictyon
Volume 2: The Proteobacteria
Phylum Proteobacteria
Class Alphaproteobacteria Agrobacterium, Caulobacter, Ehrlichia, Nitrobacter, Rhodospirillum, Rickettsia,
Rhizobium
Class Betaproteobacteria Neisseria, Nitrosomonas, Thiobacillus
Class Gammaproteobacteria Azotobacter, Chromatium, Escherichia, Legionella, Nitrosococcus, Pseudomonas,
Vibrio
Class Deltaproteobacteria Bdellovibrio, Myxococcus
Class Epsilonproteobacteria Campylobacter, Helicobacter
Volume 3: The Firmicutes
Phylum Firmicutes
Class I. Bacilli Bacillus, Streptococcus, Listeria, Staphylococcus
Class II. Clostridia Clostridium, Heliobacterium
Class III. Erysipelotrichia Erysipelotrichia
Volume 4: The Bacteroides, Spirochaetes, Tenericutes, Acidobacteria, Fibrobacteres, Fusobacteria, Dictyoglomi, Gemmatimonadetes,
Lentisphaerae, Verrucomicrobia, Chlamydiae, and Planctomycetes
Phylum Bacteroidetes Bacteroides
Phylum Spirochaetes Borrelia, Treponema
Phylum Tenericutes Mycoplasma
Phylum Acidobacteria Acidobacterium
Phylum Fibrobacteres Fibrobacter
Phylum Fusobacteria Fusobacterium
Phylum Dictyoglyomi Dictyoglomus
Phylum Gemmatimonadetes Gemmatinonas
Phylum Lentisphaerae Lentisphaera
Phylum Verrucomicrobia Verrucomicrobium
Phylum Chlamydiae Chlamydia
Phylum Planctomycetes Planctomyces
Volume 5: The Actinobacteria
Phylum Actinobacteria Corynebacterium, Bifidobacterium, Micrococcus, Mycobacterium, Streptomyces
 Part II The Microbial World 241

Bergey’s Manual of Systematic Bacteriology Microbiologists MicroAssessment 10.1

generally rely on the classifications listed in the reference text
Bergey’s Manual of Systematic Bacteriology. All known species Taxonomy consists of three interrelated areas: identification,
classification, and nomenclature. In clinical laboratories, identifying
are described there, including those that have not yet been culti- the genus and species of an isolate is more important than
vated. If the properties of a newly isolated organism do not agree understanding its evolutionary relationship to other organisms.
with any description in Bergey’s Manual, then presumably a
1. Why might it be easier to determine the cause of pneumonia than
new organism has been isolated. The newest edition of this com- the cause of a wound infection?
prehensive manual is being published in five volumes and classi-
2. What is Bergey’s Manual?
fies prokaryotes according to the most recent information on their
3. Some biologists have been reluctant to accept the three-domain
phylogeny (table 10.3). In some cases, this classification differs
system. Why might this be the case? +
substantially from that of the previous edition, which grouped
organisms according to their phenotypic characteristics.
In addition to containing descriptions of organisms, all vol-
umes include information on the ecology, methods of enrichment, 10.2 ■ Using Phenotypic
culture, and isolation of the organisms as well as methods for their
maintenance and preservation. However, the heart of the work is
Characteristics to
a description of all characterized prokaryotes and their groupings. Identify Prokaryotes
Learning Outcome
Nomenclature 2. Describe how phenotypic characteristics—including microscopic
morphology, culture characteristics, metabolic capabilities,
Bacteria are given names according to a set of internationally serology, and fatty acid analysis—can be used to identify
recognized rules, the International Code of Nomenclature of prokaryotes.
Bacteria. The names may originate from any language, but must
include a Latin suffix. In some cases, the name reflects the organ- Phenotypic characteristics can be used to help identify micro-
ism’s habitat or other characteristic but often honors a researcher. organisms, without the need for sophisticated equipment. These
Just as classification is always in a state of flux, so is the and other identification methods are summarized in table 10.4.
assignment of names. Although revising names may increase
scientific accuracy, it often leads to confusion, particularly when
names of medically important bacteria are changed. To ease the Microscopic Morphology
transition after a name change, the former name may be included An important initial step in identifying a microorganism is to
in parentheses. For example, Lactococcus lactis, a bacterium that determine its size, shape, and staining characteristics. Microscopic
was once included in the genus Streptococcus, is sometimes indi- examination gives information very quickly and is sometimes
cated as Lactococcus (Streptococcus) lactis. enough to make a presumptive identification.

TABLE 10.4 Methods Used to Identify Prokaryotes

Method Comments

Phenotypic Characteristics Most of these methods do not require sophisticated equipment and can easily be done anywhere
in the world.
Microscopic morphology Size, shape, and staining characteristics such as Gram stain can give suggestive information as to the
identity of the organism. Further testing, however, is needed to confirm the identification.
Culture characteristics Colony morphology can give initial clues to the identity of an organism.
Metabolic capabilities A battery of biochemical tests can be used to identify a microorganism.
Serology Proteins and polysaccharides that make up a prokaryote are sometimes characteristic enough to be
considered identifying markers. These can be detected using specific antibodies.
Fatty acid analysis Cellular fatty acid composition can be used as an identifying marker and is analyzed by gas
chromatography.
Genotypic Characteristics These methods are increasingly being used to identify microorganisms. Even an organism that occurs in
very low numbers in a mixed culture can be identified.
Detecting specific nucleotide Nucleic acid probes and nucleic acid amplification tests can be used to identify prokaryotes grown in
sequences culture. In some cases, the method is sensitive enough to detect the organism directly in a specimen.
Sequencing rRNA genes This requires amplifying and then sequencing rRNA genes, but it can be used to identify organisms that
have not yet been grown in culture.
242 Chapter 10 Identification and Classification of Prokaryotic Organisms

Size and Shape Streptococcus Neisseria

pneumoniae gonorrhoeae
The size and shape of a microorganism can easily be determined
by microscopically examining a wet mount. Based only on the size
and shape, the organism can be identified as a prokaryote, fungus,
or protozoan. In a clinical lab, this can sometimes be enough to
diagnose certain eukaryotic infections. For example, a wet mount
of vaginal secretions is routinely used to diagnose yeast infections,
and one of stool is examined for the eggs of parasites when certain
roundworms are suspected (figure 10.2). wet mount, p. 43

Gram Stain
The Gram stain distinguishes between Gram-positive and Gram-
negative bacteria (see figure 3.14). This relatively rapid test nar- White blood cell
rows the list of possible identities of an organism, an essential step (a) (b)
in the identification process. Gram stain, p. 47
FIGURE 10.3 Gram Stains of Clinical Specimens (a) Sputum
In a clinical lab, the Gram stain of a specimen is generally not showing Gram-positive Streptococcus pneumoniae and (b) male ure-
sensitive or specific enough to diagnose the cause of most infections, thra secretions showing Gram-negative Neisseria gonorrhoeae inside
but it is still an extremely useful tool. The technician can see the Gram white blood cells.
reaction, the shape and arrangement of the bacterial cells, and whether ? What disease does the patient in (a) have? What disease does the
the organisms appear to be growing as a pure culture or with other patient in (b) have?
bacteria and/or cells of the host. However, most medically impor-
tant bacteria cannot be identified by Gram stain alone. For example,
Streptococcus pyogenes, the bacterium that causes strep throat, cannot infection caused by Neisseria gonorrhoeae (figure  10.3b). This
be distinguished microscopically from streptococci that are part of the diagnosis can be made because N. gonorrhoeae is the only Gram-
normal throat microbiota. A Gram stain of a stool specimen cannot negative diplococcus found within white blood cells in the male
distinguish Salmonella from E. coli. These organisms generally must urethra. pneumonia, p. 486 gonorrhea, p. 622
be isolated in pure culture and tested for their biochemical capabilities
for accurate identification. strep throat, p. 487 Special Stains
In certain cases, the Gram stain result gives enough informa- Certain microorganisms have unique characteristics that can be
tion to start appropriate antimicrobial therapy. For example, a detected with special staining procedures. As an example, members
Gram stain of sputum showing numerous white blood cells and of the genus Mycobacterium are some of the few acid-fast micro-
Gram-positive diplococci is highly suggestive of Streptococcus organisms (see figure 3.15). If a patient has symptoms of tuberculo-
pneumoniae, a bacterium that causes pneumonia (figure 10.3a). sis, then an acid-fast stain will be done on a sputum sample to help
In some other cases, the Gram stain result is enough for diagnosis. identify Mycobacterium tuberculosis. acid-fast stain, p. 48
For instance, the presence of Gram-negative diplococci clustered
in white blood cells in a sample of a urethral secretion from a man
is considered diagnostic for gonorrhea, the sexually transmitted Culture Characteristics
Colony morphology can give initial clues to the identity of the
organism. For example, colonies of streptococci are generally
fairly small relative to many other types of bacteria. Colonies of
Candida albicans Roundworm egg Serratia marcescens are often red when incubated at 22°C due
to the production of a pigment. Pseudomonas aeruginosa often
produces a soluble greenish pigment, which discolors the growth
medium (see figure 11.12). In addition, cultures of P. aeruginosa
have a distinct fruity odor. growth of colonies, p. 88
In a clinical lab, where rapid but accurate diagnosis is
essential, specimens are inoculated onto differential media as a
preliminary step in the identification process. A specimen taken
by swabbing the throat of a patient complaining of a sore throat
is inoculated onto blood agar. This makes it possible to detect the
characteristic β-hemolytic colonies of Streptococcus pyogenes,
the cause of strep throat (see figure  4.10). Urine collected from
(a) (b) a patient suspected of having a urinary tract infection (UTI) is
FIGURE 10.2 Wet Mounts of Clinical Specimens (a) Vaginal plated onto MacConkey agar. E. coli, the most common cause of
secretions containing yeast (Candida albicans, 410×); (b) roundworm UTIs, ferments lactose and therefore forms pink colonies on that
(Ascaris) eggs in a stool (400×). agar (see figure  4.11). differential media, p.  95 blood agar, p.  95
? Yeast and roundworms belong to which domain? MacConkey agar, p. 95
 Part II The Microbial World 243

(a) (b) (c)

FIGURE 10.4 Biochemical Tests (a) Catalase production (left) and a negative control (right). (b) Sugar fermentation (left), negative
control (center), and an uninoculated tube (right). (c) Urease production (left), negative control (center), and an uninoculated control (right).
? What specifically causes the color to change in the sugar fermentation and urease tests?

Metabolic Capabilities certain identification (table 10.5). One of the simplest tests is an

assay for the enzyme catalase (figure 10.4a). Most bacteria that
The metabolic capabilities such as the types of sugars fermented
grow in the presence of O2 are catalase-positive. Important excep-
or the end products made can be used to identify a microorganism.
tions are the lactic acid bacteria, which include members of the
genus Streptococcus. Thus, if a throat culture yields β-hemolytic
Biochemical Tests colonies but further testing reveals they are all catalase-positive,
Colony morphology can give clues as to the identity of an organ- then Streptococcus pyogenes has been ruled out. catalase, p.  90
ism, but biochemical tests are generally necessary for a more Streptococcus pyogenes, p. 487

TABLE 10.5 Characteristics of Some Important Biochemical Tests

Biochemical Test Principle of the Test Positive Reaction

Catalase Detects the activity of the catalase, an enzyme that breaks The reagent bubbles.
down hydrogen peroxide to form O2 and water.
Citrate Determines whether or not citrate can be used as a sole Growth, usually accompanied by the color change
carbon source. of a pH indicator.
Gelatinase Detects enzymatic breakdown of gelatin to polypeptides. The solid gelatin is converted to liquid.
Hydrogen sulfide Detects H2S released as sulfur-containing amino acids are A black precipitate forms due to the reaction of
production degraded. H2S with iron salts in the medium.
Indole Detects the enzymatic removal of the amino group from The product, indole, reacts with an added chemical
tryptophan. reagent, turning the reagent a deep red color.
Lysine decarboxylase Detects the enzymatic removal of the carboxyl group from The medium becomes more alkaline, causing a pH
lysine. indicator to change color.
Methyl red Detects mixed acids, the characteristic end products of a The medium becomes acidic (pH below 4.5); a
particular fermentation pathway. mixed acids, p. 148 red color develops upon the addition of a pH
indicator.
Oxidase Detects the activity of cytochrome c oxidase, a component A dark color develops after a specific reagent is
of the electron transport chain of specific organisms. added.
cytochrome c, p. 143

Phenylalanine Detects the enzymatic removal of the amino group from The product of the reaction, phenylpyruvic acid,
deaminase phenylalanine. reacts with ferric chloride to give the medium a
green color.
Sugar fermentation Detects the acidity resulting from fermentation of the sugar The medium becomes acidic, causing a pH
incorporated into the medium; also detects gas production. indicator to change color. An inverted tube traps
any gas that is made.
Urease Detects the enzymatic degradation of urea to carbon The medium becomes alkaline, causing a pH
dioxide and ammonia. indicator to change color.
Voges-Proskauer Detects acetoin, an intermediate of the fermentation A red color develops after chemicals that detect
pathway that leads to 2,3-butanediol production. acetoin are added.
2,3-butanediol, p. 148
244 Chapter 10 Identification and Classification of Prokaryotic Organisms

Most biochemical tests rely on a chemical indicator that

changes color when a compound is degraded. To test for
the ability of an organism to ferment a given sugar, a broth
medium containing that sugar and a pH indicator is used. (a)
Fermentation of the sugar results in acid production, which
lowers the pH, resulting in a color change; an inverted tube
traps any gas produced (figure 10.4b). A medium designed to
detect urease, an enzyme that degrades urea to produce car-
bon dioxide and ammonia, contains urea and a pH indicator
(figure 10.4c).
The basic strategy for identifying bacteria based on biochemi-
cal tests relies on a dichotomous key, a flowchart of tests that
give either a positive or negative result (figure  10.5). Because
each test often requires an incubation period, however, it would
be too time-consuming to proceed one step at a time. In addition,
relying on a single biochemical test at each step could lead to
misidentification. For example, if a strain that normally gives a
positive result for a certain test loses the ability to produce a key (b)
enzyme, it would instead have a negative result. Therefore, simul- FIGURE 10.6 Commercial Modifications of Traditional
taneously inoculating multiple tests identifies the organism faster Biochemical Tests (a) An API test strip. Each small cup contains
a dried medium similar in formulation to the traditional tests. A
and more conclusively.
liquid suspension of the isolated test bacterium is added to each
In certain cases, biochemical testing can be done with- compartment; after incubation, the results are read manually.
out culturing the organism. Helicobacter pylori, the cause of (b) An Enterotube II. Each compartment contains a different type of
most stomach ulcers, can be detected using the breath test, medium. One end of a metal rod that runs through the tube is used
which assays for the presence of urease. The patient drinks a to touch a bacterial colony. When the rod is withdrawn, it inoculates
each of the compartments. After incubation, the results are read
solution containing urea labeled with an isotope of carbon. If
manually.
H. pylori is present, its urease breaks down the urea, releasing
labeled CO2, which escapes through the airway. Several hours ? What advantage do these commercial methods have over
traditional biochemical tests?
after drinking the solution, the patient exhales into a balloon,
and that air is then tested for labeled CO2. This test is less
invasive and, consequently, much cheaper and faster than the
each compartment, inoculating as well as rehydrating the media.
stomach biopsy that would be needed to culture the organism.
The media are similar to those used in traditional tests, giving
Helicobacter pylori, p. 578 isotope, p. 18
rise to comparable color changes. After a 16-hour incubation of
the inoculated test strip, the results are determined by inspec-
Commercial Variations tion. The pattern of results is converted to a numerical score,
of Traditional Biochemical Tests which is then entered into a computer to identify the organism.
Several less labor-intensive commercial variations of traditional A similar system is the Enterotube II, a tube with small com-
biochemical tests are available (figure  10.6). The API system partments, each containing a different type of medium. One
uses a strip holding a series of tiny cups that contain dried end of a metal rod that runs through the tube is used to touch
media. A liquid suspension of the test bacterium is added to a bacterial colony. The other end is used to pull the inoculated

FIGURE 10.5 Dichotomous

Gram stain
Key This shows an example of
steps that can be used to identify
some of the common causes of Gram-positive coccus Gram-negative rod
urinary tract infections. Additional
tests need to be done to confirm
Catalase Oxidase test
the identity of the pathogen.
? When identifying organisms, Positive Negative Positive Negative
why are certain biochemical
tests usually initiated
simultaneously, rather than Coagulase Pseudomonas Lactose
Enterococcus sp. aeruginosa fermentation
waiting for one result before
beginning the next test? Positive Negative Positive Negative

Staphylococcus Staphylococcus E. coli or

Proteus sp.
aureus saprophyticus other coliform
 Part II The Microbial World 245

end through the tube, thereby inoculating each of the compart- MicroAssessment 10.2
ments. A system by Biolog uses a microtiter plate, a small tray
containing nearly one hundred wells, to assay simultaneously The size, shape, and staining characteristics of a microorganism give
important clues to its identity. Conclusive identification generally
an organism’s ability to use a wide variety of carbon sources. requires multiple biochemical tests. The proteins and polysaccharides
Modifications of these plates allow researchers to characterize that make up a bacterium are sometimes unique enough to be
the metabolic capabilities of microbial communities, such as considered identifying markers. Cellular fatty acid composition can be
those in soil, water, or wastewater. used as an identifying characteristic.
Highly automated systems also are available. The VITEK  2 4. How does MacConkey agar help identify the cause of a urinary
system uses a miniature card that contains multiple wells with tract infection?
different types of dried media. After a relatively short incu- 5. Describe two methods to test for the enzyme urease.
bation period, a computer then reads the growth pattern in 6. A sample must contain many microbial cells for any to be
the wells. seen by microscopic examination. Why? +

Serology 10.3 ■ Using Genotypic

The proteins and polysaccharides that make up a prokaryotic cell
are sometimes characteristic enough to be identifying markers.
Characteristics to
The most useful of these are the molecules that make up surface Identify Prokaryotes
structures including the cell wall, capsule, flagella, and pili. For
example, some species of Streptococcus contain a unique car- Learning Outcome
bohydrate as part of their cell wall, and this distinguishes them 3. Describe how nucleic acid probes, NAATs, and sequencing 16S
from other species. These carbohydrates, as well as other distinct rRNA genes can be used to identify prokaryotes.
surface molecules, can be detected using antibodies. Detection
methods that use antibodies are called serological tests and will Many of the technologies discussed in chapter 9 can be used to
be discussed in more detail in chapter 18. Some serological identify a microorganism based on it genotype. Some of these
tests, such as those used to confirm the identity of S. pyogenes, methods even make it possible to identify organisms that cannot
are quite simple and rapid. antibodies, p.  359 antigens, p.  359 yet be grown in culture.
serology, p. 426
Detecting Specific Nucleotide Sequences
Nucleic acid probes and nucleic acid amplification tests (NAATs)
Fatty Acid Analysis (FAME) can both detect nucleotide sequences unique to a given species or
Prokaryotic species differ in the type and relative amounts of related group. A significant limitation, however, is that each probe
different fatty acids in their membranes, so fatty acid composi- or amplification detects only a single possibility. If the organ-
tion can be used as an identifying marker. To do this, bacterial ism in question could be one of five different species or related
cells are grown under standardized conditions and then treated groups, then five distinguishable probes or amplifications would
with chemicals to release the fatty acids and convert them to be needed. DNA probe, p. 232
their more volatile methyl ester form (FAME stands for fatty
acid methyl ester). The resulting FAMEs can then be separated Nucleic Acid Probes
and analyzed using gas chromatography. The chromatogram A nucleic acid probe can locate a nucleotide sequence that charac-
(pattern of peaks) is then compared to those of known species. terizes a particular species or group (figure 10.7). The probe is a

Unknown organism
(double-stranded DNA)

If probe does not bind to DNA,

Denatured then unknown organism
Probe is added to denatured, is not organism X.
Organism X Probe single-stranded DNA of
DNA unknown organism.

Double-stranded DNA is labeled, Single-stranded DNA If probe binds to DNA,

DNA sequence then denatured then unknown organism
unique to to become is organism X.
organism X a probe.

FIGURE 10.7 Nucleic Acid Probes to Detect Specific DNA Sequences The probe, a single-stranded piece of nucleic acid labeled with a
detectable marker, is used to locate a unique nucleotide sequence that identifies a particular microbial species.
? What type of label does the probe used in fluorescence in situ hybridization (FISH) employ?
246 Chapter 10 Identification and Classification of Prokaryotic Organisms

single-stranded piece of nucleic acid, usually DNA, labeled with

a detectable tag such as a radioisotope or a fluorescent dye. It is
complementary to the sequence of interest. 70S
Most methods that use nucleic acid probes to detect DNA
sequences rely on a step that increases the amount of DNA in
the sample. This can be done by inoculating the specimen onto
an agar medium so that each microbial cell multiplies, forming a
colony. Alternatively, a preliminary in vitro DNA amplification
step can be done.
Fluorescence in situ hybridization (FISH) often uses probes 30S 50S
that bind 16S ribosomal RNA (rRNA). An amplification step is
not needed because numerous copies of rRNA are naturally pres-
ent in multiplying cells. Various different probes that bind rRNA
are available, each specific for a given signature sequence. A
signature sequence is a sequence in rRNA that characterizes either
a certain species or a group of related organisms (see figure 9.20).
Applications of FISH, as well as the techniques, were discussed in
chapter 9. fluorescence in situ hybridization (FISH), p. 232 16S rRNA 5S rRNA
+ +
21 polypeptide chains 23S rRNA
Nucleic Acid Amplification Tests (NAATs) +
Several methods, referred to as nucleic acid amplification 34 polypeptide chains
tests (NAATs), can be used to increase the number of copies of FIGURE 10.8 Ribosomal RNA The 70S ribosome of prokaryotes
specific DNA sequences. This allows researchers to detect spe- has three types of rRNA: 5S, 16S, and 23S.
cific sequences in samples such as body fluids, soil, food, and ? Which type of ribosomal RNA is most often used in taxonomy?
water. These methods can be used to detect organisms present
in extremely small numbers as well as those that cannot yet be
grown in culture. In most cases, the amplified fragment is visible
as a distinct band on an ethidium bromide–stained agarose gel Using rDNA to Identify
illuminated with UV light. Alternatively, a DNA probe can be Uncultivated Organisms
used to detect the amplified DNA. ethidium bromide, p. 218 gel The vast majority of microbes cannot yet be grown in culture.
electrophoresis, p. 217 However, the DNA from these organisms can be amplified,
One of the most common NAATs is the polymerase chain cloned, and sequenced, making it possible to detect and identify
reaction (PCR), described in chapter 9 (see figure  9.14). To use them. The bacterium that causes Whipple’s disease, a rare intes-
PCR to detect a microbe of interest, a sample is treated to release tinal illness, was identified this way. The organism was given
and denature the DNA. Specific primers and other ingredients are the name Tropheryma whipplei, and a specific probe was then
then added (see figure  9.15). After 30 cycles of PCR, the target developed to detect it in intestinal tissue, well before it could be
DNA will have been amplified approximately a billion-fold (see grown in culture.
figure 9.17). PCR, p. 227
MicroByte
A gram of fertile soil may contain over 4,000 species of prokaryotes,
Sequencing Ribosomal RNA Genes most of which have not been identified.
The nucleotide sequence of ribosomal RNA molecules (rRNAs),
or the DNA that encodes them (rDNAs), can be used to identify
prokaryotes (figure  10.8). rRNAs are useful in microbial clas- MicroAssessment 10.3
sification and identification because their sequences are relatively
A microorganism can be identified by using a probe or NAAT to
stable; the ribosome would not function with too many mutations. detect a nucleotide sequence unique to that organism. Ribosomal
Of the different rRNAs (5S, 16S, and 23S), the 16S molecule is RNA genes can be sequenced to identify organisms, including those
the most useful in taxonomy because of its moderate size (approxi- that cannot be grown in culture.
mately 1,500 nucleotides). 16S RNA and its eukaryotic counterpart, 7. When using a probe to detect an organism, why is it necessary to
18S RNA, are small subunit (SS, or SSU) rRNAs, meaning they are have some idea as to the organism’s identity?
part of the small subunit of the ribosomes (figure 10.8). Once the 8. Why is 16S RNA also referred to as SSU RNA?
nucleotide sequence of an unknown organism’s SSU RNA has been
9. Why are molecular methods especially useful when bacteria are
determined, it can be compared with sequences of known organisms difficult to grow? +
by searching extensive computerized databases.
 Part II The Microbial World 247

10.4 ■ Characterizing Strain

Differences
Flagella
(H antigen)
Learning Outcome
4. Describe five distinct methods to distinguish different strains.

In some situations, distinguishing different strains of a given spe-

cies is useful. In 2009, for example, 235 reported salmonellosis
cases across 14 states in the United States involved a specific strain
Capsule
of Salmonella enterica serotype Saintpaul. Most patients reported (K antigen)
consuming raw alfalfa sprouts prior to their illness, leading to a
recall of the seeds by the producer. Linking 235 salmonellosis
cases amid the thousands that occur nationwide each year would
be impossible without methods to distinguish different strains.
Salmonella, p. 592
Cell wall
Characterizing strain differences is not limited to investi- (O antigen of
gations of foodborne illness. It also plays an important role in outer membrane)
forensic investigations of bioterrorism and other biocrimes, and
in diagnosing certain diseases. The methods used to characterize FIGURE 10.9 Serotypes The cell structures used to distinguish dif-
different strains are summarized in table 10.6. ferent strains of members of the family Enterobacteriacea are shown.
? What structures are reflected in the “O157:H7” of E. coli O157:H7?
Biochemical Typing
Biochemical tests are used to identify various species of bacteria, O157:H7 refers to the antigenic type of its lipopolysaccharide
but they can also be used to distinguish strains. A group of strains (the O antigen) and flagella (the H antigen). A group of strains
that have a characteristic biochemical pattern is called a biovar, that have cell surface antigens different from other strains is
or a biotype. A biochemical variant of Vibrio cholerae called called a serovar, or a serotype. antigen, p. 359 lipopolysaccha-
El Tor caused a worldwide epidemic of cholera beginning in ride, p. 60 E. coli O157:H7, p. 590
1961. Because this biovar can be readily distinguished from other
strains, its spread can be traced. Vibrio cholerae, p. 586
Molecular Typing
Subtle differences in DNA sequences can be used to distinguish
Serological Typing among phenotypically identical strains, and these genomic varia-
Proteins and carbohydrates that vary among strains can be used tions are important for tracing epidemics of foodborne illness. One
as distinguishing markers. For example, different strains of method of doing this is to compare the patterns of fragment sizes
E. coli and related bacteria can be distinguished by the anti- produced when the same restriction enzyme is used to digest DNA
genic type of their flagella, capsules, and lipopolysaccharide from each isolate; the different patterns are called restriction
molecules (figure 10.9). The “O157:H7” designation of E. coli fragment length polymorphisms (RFLPs). Gel electrophoresis

TABLE 10.6 Summary of Methods Used to Characterize Different Strains

Method Comment

Biochemical typing Biochemical tests are most commonly used to identify various species of bacteria, but in some cases they can
be used to distinguish different strains. A group of strains that have a characteristic biochemical pattern is
called a biovar or a biotype.
Serological typing Proteins and carbohydrates that vary among strains can be used to differentiate strains. A group of strains
that have a characteristic serological type is called a serovar or a serotype.
Molecular typing Molecular methods such as pulsed-field gel electrophoresis can be used to detect restriction fragment length
polymorphisms (RFLPs). Multilocus sequencing typing compares certain nucleotide sequences.
Phage typing Strains of a given species sometimes differ in their susceptibility to various types of bacteriophage.
Antibiograms Antibiotic susceptibility patterns can be used to characterize strains.
248 Chapter 10 Identification and Classification of Prokaryotic Organisms

is used to separate the fragments so they can be observed; if the foodborne bacterial pathogens. Laboratories from around the
fragments are quite large, a special method called pulsed-field gel country can submit RFLP patterns to a computer database and
electrophoresis (PFGE) is used. Isolates of the same species that quickly receive information about other isolates showing the same
have different RFLPs are considered different strains, whereas pattern. Using this database, multistate foodborne disease out-
those have identical RFLPs may or may not be the same strain breaks can more readily be recognized and traced. This is how the
(figure 10.10). restriction enzymes, p. 315 salmonellosis cases that led to the alfalfa seeds recall were found
To make it easier to track foodborne disease outbreaks, the to be related (see Perspective 10.1). CDC, p. 446
CDC established PulseNet, which catalogs the RFLPs of certain A newer method for distinguishing different strains is
multilocus sequence typing (MLST). Automated sequencing
Restriction sites methods are used to determine the nucleotide sequences of por-
tions of seven or so common genes, and these sequences are then
Strain A Cut with restriction enzyme compared.
DNA

3 kb 6 kb 1 kb
Phage Typing
Strains of a given species may differ in their susceptibility to
bacteriophages. Bacteriophages, or phages, are viruses that infect
Strain B
DNA bacteria, often lysing them; they will be described in more detail
in chapter 13. The susceptibility of an organism to a particular
type of phage can be easily determined. First, a culture of the
5 kb 4 kb 1 kb
test organism is inoculated into melted, cooled nutrient agar
and poured onto the surface of an agar plate, creating a uniform
Strain C layer of cells. Drops of different types of bacteriophage are then
DNA
placed on the surface of the agar. During incubation, the bacteria
multiply, forming a visible haze of cells. If the bacterial strain is
3 kb 2 kb 4 kb 1 kb susceptible to a specific type of phage, a clear area will form at
(a)
the spot where bacteriophage was added. The patterns of clear-
ing indicate the susceptibility of the test organism to different
phages (figure 10.11). Bacteriophage typing has now largely been
replaced by molecular methods that detect genomic differences,
but it is still a useful tool for laboratories that lack equipment to
do molecular typing. bacteriophage, p. 304

Antibiograms
Antibiotic susceptibility patterns, or antibiograms, can distin-
guish different strains. As with phage typing, this method has
largely been replaced by molecular techniques. To determine the
antibiogram, a culture is uniformly inoculated onto the surface
of nutrient agar. Paper discs containing different antibiotics are
then placed on the agar. After incubation, clear areas will be vis-
ible around discs of antibiotics that inhibit or kill the organism
(figure 10.12). antibiotic, p. 458

MicroAssessment 10.4
(b)
Strains of a given species may differ in phenotypic characteristics
such as biochemical capabilities, protein and polysaccharide
FIGURE 10.10 Restriction Fragment Length
Polymorphisms (RFLPs) (a) Different strains of a species may have components, susceptibility to bacteriophages, and sensitivity to
subtle variations in nucleotide sequences that give rise to a slightly antimicrobial drugs. Molecular techniques can be used to detect
different assortment of restriction fragment sizes (1 kb = 1,000 base genomic differences between strains that are phenotypically
pairs). (b) In the method shown, genomic DNA is digested with a identical.
restriction enzyme that cuts infrequently; the resulting fragments are 10. Explain the difference between a biotype and a serotype.
separated by pulsed-field gel electrophoresis and then stained with
ethidium bromide.
11. Describe the significance of RFLPs in distinguishing between
strains.
? How are RFLPs used to track foodborne disease outbreaks?
 Part II The Microbial World 249

1 An inoculum of Staphylococcus aureus is spread over the surface

of agar medium.

Inoculum of
Staphylococcus
aureus strain
Agar medium to be typed

Petri dish
FIGURE 10.12 An Antibiogram In this example, 12 different
antimicrobial drugs incorporated in paper discs have been placed on
two plates containing different cultures of Staphylococcus aureus.
Clear areas represent zones of inhibited growth.
? How can you tell that these S. aureus isolates are different strains?

10.5 ■ Classifying Prokaryotes

2 Different bacteriophage suspensions are deposited in a fixed pattern. Learning Outcome
5. Describe how 16S rRNA sequences, DNA hybridization, and DNA
base ratios are used to classify prokaryotes.

The goal of phylogenetic classification is to group organisms

according to their evolutionary relatedness. Unfortunately, this is
difficult when trying to place the diverse types of prokaryotes in
their proper positions with respect to evolution. Fossilized stro-
matolites (coral-like mats of filamentous microorganisms) can
be studied, but these remains give few clues to help identify or
understand ancient microbes. Because evolutionary relatedness
can be difficult to determine, prokaryotic classification schemes
historically grouped organisms by their phenotypic traits such as
size and shape, staining characteristics, and metabolic capabilities.
3 After incubation, different patterns of lysis are seen with different Although this is easy, there are several significant drawbacks.
strains of S. aureus. For instance, phenotypic differences can be due to only a few
Dye marker gene products, and a single mutation resulting in a non-functional
enzyme can change an organism’s capabilities. In addition,
phenotypically similar organisms may be only distantly related;
conversely, those that appear dissimilar may be closely related.
Newer molecular techniques such as DNA sequencing avoid
some of the problems associated with phenotypic classification
while also giving greater insights into the evolutionary relatedness
of microorganisms. DNA sequences are viewed as evolutionary
chronometers, meaning they provide a relative measure of the
time elapsed since the organisms diverged from a common ances-
tor. This is because random mutations cause sequences to change
over time. The more time elapsed since two organisms diverged,
the greater the differences in the sequences of their DNA.
DNA sequencing makes it possible to more accurately
Lysis construct a phylogenetic tree. These trees are somewhat like a
family tree, tracing the evolutionary heritage of organisms. Each
FIGURE 10.11 Phage Typing branch represents the evolutionary distance between two species,
? Why would a lab do phage typing rather than molecular typing? and individual species are represented as nodes (figure  10.13).
250 Chapter 10 Identification and Classification of Prokaryotic Organisms

Aquifex pyrophilus Aquificae

Thermotoga maritima Thermotogae
Deinococcus radiodurans “Deinococcus-Thermus”
Thermus aquaticus
Chloroflexus aurantiacus Chloroflexi
Corynebacterium glutamicum
Mycobacterium tuberculosis
Micrococcus luteus Actinobacteria
(High G + C Gram-positives)
Streptomyces griseus
Frankia sp.
Fusobacterium ulcerans Fusobacteria
Staphylococcus aureus
Bacillus cereus
Enterococcus faecalis Firmicutes
Streptococcus pyogenes (Low G + C Gram-positives)
Mycoplasma pneumoniae
Clostridium perfringens
Anabaena “cylindrica”
Synechococcus lividus Cyanobacteria
Oscillatoria sp.
Chlamydia trachomatis Chlamydiae
Planctomyces maris Planctomycetes
Chlorobium limicola Chlorobi
Flexibacter litoralis
Cytophaga aurantiaca Bacteroidetes
Flavobacterium hydatis
Bacteroides fragilis
Fibrobacter succinogenes Fibrobacteres
Treponema pallidum Spirochaetes
Borrelia burgdorferi
Campylobacter jejuni Epsilonproteobacteria
Helicobacter pylori
Desulfovibrio desulfuricans
Bdellovibrio bacteriovorus Deltaproteobacteria
Myxococcus xanthus
Rickettsia rickettsii
Caulobacter crescentus Alphaproteobacteria
Rhodospirillum rubrum Proteobacteria
Vibrio cholerae
Escherichia coli Gammaproteobacteria
Pseudomonas aeruginosa
Neisseria gonorrhoeae
Alcaligenes denitrificans Betaproteobacteria
Nitrosococcus mobilis
FIGURE 10.13 Phylogenetic Tree of the Bacteria Each branch represents the evolutionary distance between two species.
? Which is more closely related—Deinococcus radiodurans and Thermus aquaticus or Bacillus cereus and Clostridium perfringens?

Ancient prokaryotes, which branch early in evolution, are called Bacteria Eucarya Archaea
deeply branching to reflect their position in the phylogenetic tree.
Although analysis of sequence data has solved some
Proteobacteria

Cyanobacteria

Animalia

Fungi

Plantae

Euryarchaeota

Crenarchaeota
difficulties in prokaryotic classification, it has also highlighted an
important obstacle. Prokaryotic cells transfer DNA to other spe-
cies, a process called horizontal gene transfer, complicating
Archezoa

insights provided by DNA sequence comparison. The bacterium

Thermotoga maritima, for example, appears to have acquired one- sts
ro pla
fourth of its genes from a hyperthermophilic archaeal species.
C hlo
Such observations have prompted some scientists to suggest that ia
n dr
the tree of life (see figure 10.1) should be depicted as a shrub with ho
to c
interwoven branches (figure 10.14). horizontal gene transfer, p. 200 Mi
Table 10.7 summarizes some of the methods used to classify
prokaryotes.

16S rDNA Sequence Analysis

Analyzing and comparing the nucleotide sequences of 16S FIGURE 10.14 “Shrub” of Life
ribosomal RNA (rRNA) and the genes that encode rRNA ? Why are chloroplasts and mitochondria depicted as arrows
(rDNA) have revolutionized the classification of organisms originating from the Bacteria?
 Part II The Microbial World 251

PERSPECTIVE 10.1
Tracing the Source of an Outbreak of Foodborne Disease
On September 8, 2006, Wisconsin state health intestine and can be found in almost every E. coli colonies are serotype O157; those that
authorities alerted the CDC about an E. coli sample of feces. How was this particular strain test positive are then generally tested to con-
O157:H7 outbreak that appeared to involve separated and distinguished from the hundreds firm they are serotype H7.
consumption of fresh spinach. Soon thereafter, of E. coli strains that do not cause diarrheal The next task is to determine whether or
Oregon officials reported a similar outbreak. disease? It is also true that sporadic cases of not two isolates of E. coli O157:H7 originated
On September 12, the CDC determined that E. coli O157:H7 infections occur regularly, from the same source. DNA is extracted and
the strains from Wisconsin matched those from originating from unrelated sources. How was purified from each isolate and is then digested
Oregon, as well as strains then reported from New it recognized that these cases were connected? with restriction enzymes. Pulsed-field gel elec-
Mexico. Within days, the CDC issued a press To identify E. coli O157:H7 in a stool trophoresis is generally used to compare the
release advising people not to eat bagged fresh specimen, the sample is inoculated onto resulting restriction fragment length polymor-
spinach, and a California company that produces a special agar medium designed to distin- phism (RFLP) patterns of the isolates (see
several brands of bagged spinach announced a guish it from strains that typically inhabit the figure  10.10). Those that have identical pat-
voluntary recall of all fresh spinach products. In large intestine. One such medium is sorbitol- terns are presumed to have originated from the
the end, the implicated strain was found to have MacConkey, a modified version of MacConkey same source. Patients from whom those isolates
caused 205 cases of illness in 26 states, resulting agar in which the lactose is replaced with the originated can then be questioned to deter-
in three deaths. E. coli O157:H7, p. 590 carbohydrate sorbitol. On this medium, most mine where they likely contacted the disease-
Although the sequence of events that led E. coli O157:H7 isolates are colorless because causing organism. Culture methods are then
to the recognition of the E. coli O157:H7 they do not ferment sorbitol. In contrast, com- used to try to isolate the organism from the
outbreak may sound quite simple, they are mon strains of E. coli ferment the carbohy- suspected food source. If that attempt is suc-
actually very complex. For one thing, most drate, giving rise to pink colonies. Serology cessful, the RFLP pattern of that isolate is then
strains of E. coli are normal inhabitants of the is then used to determine if the colorless compared with those of the related cases.

(see A Glimpse of History). This is because rRNA is pres- as soil and water, and the 16S rDNA then amplified, cloned, and
ent in all organisms and performs a critical and functionally sequenced. The sequences can then be compared to databases
constant task. The number of mutations that can happen in containing 16S rDNA sequences of known organisms. Using
certain regions without affecting the survival of an organism is these techniques, a variety of unique prokaryotes have been dis-
limited. Long after organisms have diverged, portions of their covered. In fact, even though most characterized archaeal genera
16S rDNA sequences are still similar. Changes in these highly are extremophiles, 16S rDNA sequences indicate that members of
conserved regions occur very slowly over time and therefore are this domain are common in non-extreme environments as well.
useful for determining distant relationships of diverse organisms. Clearly, the prokaryotic world is much more diverse than previ-
At the same time, certain regions are relatively variable. These ously recognized. using rDNA to identify uncultivated organisms, p. 246
sequences can be used to determine more recent divergence. In Although 16S rDNA sequence analysis has been very helpful
addition, horizontal gene transfer is unlikely to complicate the in determining the phylogeny of distantly related organisms, it is
analysis because rDNA transfer appears to be rare. often unreliable for distinguishing closely related species. This is
The phylogeny of the many prokaryotes not yet grown in because closely related but genetically distinct prokaryotes can
culture can be tentatively determined by 16S rDNA sequence have identical 16S rDNA sequences. In these cases, DNA hybrid-
analysis. DNA can be extracted from environmental samples such ization (discussed next) is a better tool to assess relatedness.

Methods Used to Determine the Relatedness of Different Prokaryotes

TABLE 10.7
for Purposes of Classification
Method Comment

Genotypic Characteristics Differences in DNA sequences can be used to determine the point in time at which two organisms diverged
from a common ancestor.
Comparing the sequences This technique has revolutionized classification. Certain regions of the 16S rDNA can be used to determine
of 16S rDNA distant relatedness of diverse organisms; other regions can be used to determine more recent divergence.
DNA hybridization The extent of nucleotide sequence similarity between two isolates can be determined by measuring how
completely single strands of their DNA hybridize to one another.
DNA base composition Determining the G + C content offers a crude comparison of genomes. Organisms with identical G + C
contents can be entirely unrelated, however.
Phenotypic Characteristics Traditionally, relatedness of different bacteria has been decided by comparing properties such as ability to
degrade lactose and the presence of flagella. These characteristics, however, do not necessarily reflect the
evolutionary relatedness of organisms.
252 Chapter 10 Identification and Classification of Prokaryotic Organisms

DNA Hybridization 1.5

Relative absorbance at 260 nm

The extent of nucleotide sequence similarity between two organ-
isms can be determined by measuring how completely single 1.4
strands of their DNA hybridize to one another. Just as the
complementary strands of DNA from one organism will anneal 1.3
(base-pair), so will homologous DNA of a different organism.
The extent of hybridization reflects the degree of sequence simi-
1.2
larity. Two strains that show at least 70% similarity are generally
considered to be members of the same species. Surprisingly,
DNA hybridization studies have shown that members of the gen- 1.1
era Shigella and Escherichia, which are quite different based on
biochemical tests, should actually be grouped in the same species. 1.0
Note that human and chimpanzee DNA have approximately 99% 70°C 80°C Tm 90°C 100°C
similarity by DNA hybridization studies. Therefore, by the criteria
used to classify prokaryotes, humans and chimpanzees would be FIGURE 10.15 A DNA Melting Curve The absorbance (relative
absorbance at 260 nm) rapidly increases as double-stranded DNA
members of the same species! DNA hybridization, p. 232
denatures (melts).
? What characteristic of a DNA sequence does the Tm reflect?
DNA Base Ratio (G + C Content)
One way to roughly compare the genomes of different bacteria is
Phenotypic Methods
to determine their DNA base ratio, which is the relative portion of
adenine (A), thymine (T), guanine (G), and cytosine (C). Because Classification schemes that group organisms by phenotype have
of base-pairing rules, the number of molecules of G in double- largely been replaced by methods that rely on 16S ribosomal
stranded DNA always equals the number of molecules of C. nucleic acid sequence data. Some taxonomists, however, believe
Likewise, the number of molecules of T equals that of A. The base classification should be based on more than just genotypic traits.
ratio of an organism is usually expressed as the percent of guanine Regardless, phenotypic methods are still important because they
plus cytosine, termed the G + C content, or more commonly, the provide a foundation for prokaryotic identification.
GC content. If the GC content of two organisms differs by more Numerical taxonomy uses a quantitative approach to phe-
than a small percentage, they cannot be closely related. A similar- notypic classification, comparing a battery of characteristics.
ity of base compositions, however, does not necessarily mean that Information about this method can be obtained by visiting the text
the organisms are related, because the nucleotide sequences and website (www.mhhe.com/nester7).
genome sizes could differ greatly.
The GC content is often measured by determining the tem- MicroAssessment 10.5
perature at which the double-stranded DNA denatures, or melts. Analyzing and comparing the sequences of ribosomal RNA genes has
DNA that has a high GC content melts at a higher temperature revolutionized the classification of organisms. Other characteristics
because G-C base pairs are held together by three hydrogen bonds, used to classify prokaryotes include DNA hybridization, GC ratio,
whereas A-T pairs are held together by only two hydrogen bonds. and phenotypic properties.
The temperature at which double-stranded DNA melts can read- 12. Explain why 16S rDNA is useful for determining phylogeny.
ily be determined by measuring the absorbance of UV light by a 13. Explain the role of DNA hybridization in classification.
solution of DNA as it is heated. The absorbance rapidly increases 14. Why would it be easier to sequence rDNA than rRNA? +
as the DNA denatures (figure 10.15).

FUTURE CHALLENGES 10.1

Tangled Branches in the Phylogenetic Tree
Although 16S rDNA sequencing has given data. This discrepancy is probably due to As the genetic relatedness among organisms
remarkable new insight into the evolution- horizontal gene transfer (HGT), which can becomes easier to determine, nomenclature
ary relatedness of organisms, total genome result in two distantly related microbes sharing is bound to change Already, some species
sequencing is revealing a tremendous amount some of the same nucleotide sequences. The formerly included in the genus Streptococcus
of additional information. In some cases, this outcome, however, is that the phylogenetic have been removed to create two relatively
helps clarify the evolutionary picture, provid- tree may be better illustrated as a shrub, with new genera, Enterococcus and Lactococcus.
ing solid evidence that two organisms are, HGT giving rise to intertwining branches (see Meanwhile, other microbial species will be
in fact, closely related. However, genomic figure 10.14). moved from one existing genus to another.
sequencing sometimes gives information that New insights with respect to microbial Any change creates a potential source of con-
conflicts with conclusions based on rDNA phylogeny also affect bacterial nomenclature. fusion for the scientist and layperson alike.
 Part II The Microbial World 253

Summary
10.1 ■ Principles of Taxonomy DNA that encodes rRNA. Organisms that cannot yet be cultivated
Taxonomy consists of three interrelated areas: identification, can be identified by amplifying, cloning, and then sequencing specific
classification, and nomenclature. regions of rDNA.
Strategies Used to Identify Prokaryotes 10.4 ■ Characterizing Strain Differences (table 10.6)
To characterize and identify microorganisms, a wide assortment of
technologies is used, including microscopic examination, cultural char- Biochemical Typing
acteristics, biochemical tests, and nucleic acid analysis. A group of strains that has a characteristic biochemical variation is
called a biovar, or a biotype.
Strategies Used to Classify Prokaryotes
Taxonomic classification categories are arranged in a hierarchical order, Serological Typing
with the species being the basic unit. Taxonomic categories include A group of strains that differs serologically from other strains is called
species, genus, order, class, phylum (or division), kingdom, and domain. a serovar, or a serotype (figure 10.9).
Individual strains within a species vary in minor properties (table 10.1).
Molecular Typing
Nomenclature Two isolates that have different restriction fragment length poly-
Prokaryotes are assigned names governed by official rules. morphisms (RFLPs) are considered different strains (figure  10.10).
Multilocus sequence typing determines the nucleotide sequence of
10.2 ■ Using Phenotypic Characteristics representative segments.
to Identify Prokaryotes (table 10.4)
Phage Typing
Microscopic Morphology The susceptibility to various types of bacteriophages can be used to
The size, shape, and staining characteristics of a microorganism yield demonstrate strain differences (figure 10.11).
important clues as to its identity (figures 10.2, 10.3).
Antibiograms
Culture Characteristics Antibiotic susceptibility patterns can be used to distinguish strains
Selective and differential media used in the isolation process can pro- (figure 10.12).
vide information that helps identify an organism.
10.5 ■ Classifying Prokaryotes (table 10.7)
Metabolic Capabilities DNA sequences can be used to construct a phylogenetic tree
Most biochemical tests rely on a pH indicator or chemical reaction (figure  10.13). Horizontal gene transfer can complicate insights
that shows a color change when a compound is degraded. The basic provided by some types of DNA sequence comparison (figure 10.14).
strategy for identification using biochemical tests relies on the use of a
dichotomous key (figures 10.4, 10.5; table 10.5). 16S rDNA Sequence Analysis
Analyzing and comparing the sequences of rRNA and more recently,
Serology rDNA, has revolutionized the classification of organisms.
Proteins and polysaccharides that make up a prokaryote’s surface are
sometimes characteristic enough to be identifying markers. DNA Hybridization
The extent of nucleotide similarity between two organisms can be deter-
Fatty Acid Analysis (FAME) mined by measuring how completely single strands of their DNA will
Cellular fatty acid composition can be used as an identifying marker. anneal to one another.

10.3 ■ Using Genotypic Characteristics DNA Base Ratio (G + C Content)

to Identify Prokaryotes The G + C content can be measured by determining the temperature at
which double-stranded DNA melts (figure 10.15).
Detecting Specific Nucleotide Sequences
A probe complementary to a sequence unique to a given microbe is Phenotypic Methods
used to detect that organism (figure  10.7). Nucleic acid amplification Classification schemes that group organisms by phenotype have largely
tests (NAATs) such as PCR increase the number of copies of a specific been replaced by methods that rely on 16S ribosomal nucleic acid
nucleotide sequence, thereby determining if a given organism is present. sequence data.
Sequencing Ribosomal RNA Genes
The nucleotide sequence of ribosomal RNA (rRNA) can be used to
identify prokaryotes. Newer techniques simply sequence rDNA, the

Review Questions
Short Answer 3. Describe how a dichotomous key is used when identifying
1. Name and describe each of the areas of taxonomy. bacteria.
2. Compare and contrast the five-kingdom and three-domain systems 4. Describe the difference between using a probe and using PCR to
of classification. detect a specific sequence.
254 Chapter 10 Identification and Classification of Prokaryotic Organisms

5. Explain how signature sequences are used in bacterial 8. Which of the molecular methods of assessing similarity gives the
identification. crudest approximation of relatedness?
6. Describe the function of PulseNet. a) DNA hybridization b) PCR
7. Describe how the GC content of DNA can be measured. c) 16S rDNA sequencing d) DNA base composition
8. Explain why DNA sequences are evolutionary chronometers. 9. The sequence of which ribosomal genes are most commonly used
9. What is a phylogenetic tree? for establishing phylogenetic relatedness?
10. Why should a classification scheme reflect the phylogeny of a) 5S b) 16S
organisms? c) 23S d) All of these are commonly used.
10. All of the following statements are correct except
Multiple Choice a) Tropheryma whipplei could be identified before it had been
1. Which of the following is the newest taxonomic unit? grown in culture.
a) Strain b) Family c) Order b) the GC content of DNA can be measured by determining the
d) Species e) Domain temperature at which double-stranded DNA melts.
2. An acid-fast stain can be used to detect which of the following c) sequence differences between organisms can be used to assess
organisms? their relatedness.
a) Cryptococcus neoformans b) Mycobacterium tuberculosis d) based on DNA homology studies, members of the genus
Shigella should be in the same species as Escherichia coli.
c) Neisseria gonorrhoeae d) Streptococcus pneumoniae
e) gel electrophoresis is used to determine the serotype of an
e) Streptococcus pyogenes
organism.
3. The “breath test” for Helicobacter pylori infection determines the
presence of which of the following? Applications
a) Antigens b) Catalase c) Hemolysis 1. Microbiologists debate the use of biochemical similarities and
d) Lactose fermentation e) Urease cell features as a way of determining the taxonomic relationships
4. The “O157:H7” of E. coli O157:H7 refers to the among prokaryotes. Explain why some microbiologists believe
a) biotype. b) serotype. c) phage type. these similarities and differences are a powerful taxonomic indica-
tor, whereas others think they are not very useful for that purpose.
d) ribotype. e) antibiogram.
2. A researcher interested in investigating the genetic relationship of
5. PulseNet catalogs which of the following?
mitochondria to bacteria must decide on the best method to study
a) Biotype b) Serotype c) Phage type this. What advice would you give the researcher?
d) RFLP e) Antibiogram
6. Which of the following is an example of an evolutionary Critical Thinking +
chronometer? 1. In figure 10.15, how would the curve appear if the GC content of
a) Ability to form endospores the DNA sample were increased? How would the curve appear if
b) 16S ribosomal RNA sequence the AT content were increased?
c) Sugar degradation 2. When DNA probes are used to detect specific sequence similarities
d) Motility in bacterial DNA, the probe is heated and the two strands of DNA
7. If the GC content of two organisms is 70%, which of the following are separated. Why must the probe DNA be heated?
is true?
a) The organisms are definitely related.
b) The organisms are definitely not related.
c) The AT content is 30%.
d) The organisms likely have extensive DNA homology.
e) The organisms likely have many characteristics in common.
11
The Diversity of
Prokaryotic Organisms
KEY TERMS
Anoxygenic Phototrophs complex structures called fruiting
Photosynthetic organisms that do not bodies.
produce O2. Nitrifiers Gram-negative bacteria
Chemolithotrophs Organisms that obtain energy by oxidizing
that harvest energy by oxidizing inorganic nitrogen compounds such
inorganic chemicals. as ammonia or nitrate.
Chemoorganotrophs Organisms Oxygenic Phototrophs
that harvest energy by oxidizing Photosynthetic organisms that
organic chemicals. produce O2.
Cyanobacteria Gram-negative Prosthecate Bacteria
oxygenic phototrophs; genetically Gram-negative bacteria that have
related to chloroplasts. extensions projecting from the
cells, thereby increasing their
Lactic Acid Bacteria Gram-
surface area.
positive bacteria that generate lactic
acid as a major end product of their Spirochetes Long helical bacteria
fermentative metabolism. that have flexible cell walls and
endoflagella.
Methanogens Archaea that obtain
energy by oxidizing hydrogen gas, Sulfur-Oxidizing Bacteria
using CO2 as a terminal electron Gram-negative bacteria that obtain
acceptor, thereby generating energy by oxidizing elemental sulfur
methane. and reduced sulfur compounds,
generating sulfuric acid.
Myxobacteria Gram-negative
Helicobacter pylori (color-enhanced transmission electron micrograph).
bacteria that congregate to form

A Glimpse of History
Cornelis B. van Niel (1897–1985) earned his Ph.D. from the Technological
University in Delft, Holland, the home of an approach to microbiology
now commonly referred to as “the Delft School.” The outstanding program bacteriology course, inspiring many microbiologists with his enthusiasm
there was chaired in succession by two well-known microbiologists— for the diversity of microorganisms and their importance in nature. His
Martinus Beijerinck and Albert Kluyver. sharp memory and knowledge of the literature, along with his apprecia-
As Kluyver’s student, van Niel was influenced by his mentor’s tion for the remarkable abilities of microbes, allowed him to convey the
belief that biochemical processes were fundamentally the same in all wonders of the microbial world to his students.
cells and that microorganisms could be important research tools, serv-
ing as a model to study biochemical process. Thirty years later, Kluyver

S
and van Niel presented lectures that would be published in the book cientists are only beginning to understand the vast diversity
The Microbe’s Contribution to Biology. of microbial life. Although a million species of prokaryotes
Shortly after completing his dissertation in 1928, van Niel accepted a are thought to exist, only approximately 6,000 of these,
position at the Hopkins Marine Station in California. There, he continued grouped into over 950 genera, have been described and classified.
work he started under Kluyver’s direction on the photosynthetic activi- Traditional culture and isolation techniques have not supported
ties of the vividly colored purple bacteria. He demonstrated that these the growth and subsequent study of the vast majority. This situ-
microbes require light for growth, yet, unlike plants and algae, do not ation is changing as new molecular techniques make it easier to
evolve O2. He also showed that to fix CO2, the purple bacteria oxidize discover and characterize previously unrecognized species. The
hydrogen sulfide. Furthermore, van Niel noted that the photosynthetic
sheer volume of information uncovered by modern technologies,
reactions in all photosynthetic organisms are remarkably similar, except
the purple bacteria use hydrogen sulfide in place of water and produce
however, can be daunting for scientists and students alike.
oxidized sulfur compounds instead of O2. This finding raised the pos- This chapter covers a variety of prokaryotes, focusing primar-
sibility that O2 generated by plants and algae did not come from carbon ily on their extraordinary diversity rather than the phylogenetic
dioxide, as was believed at the time, but rather from water. relationships discussed in chapter 10. Note, however, that no
In addition to his scientific contributions, van Niel was an outstand- single chapter could describe all known prokaryotes and, conse-
ing teacher. During the summers at Hopkins Marine Station, he taught a quently, only a relatively small selection is presented.

255
256 Chapter 11 The Diversity of Prokaryotic Organisms

METABOLIC DIVERSITY
As a group, prokaryotes use an impressive array of mechanisms This process generates methane (CH4), a colorless, odorless, flam-
to harvest energy in order to produce ATP. This section will high- mable gas:
light this metabolic diversity by describing select prokaryotes.
4 H2 + CO2 → CH4 + 2 H2O
Table 11.1 summarizes characteristics of the archaea and bacteria
(energy source) (terminal electron
covered in this section.
acceptor)
Many methanogens can also use alternative energy sources
11.1 ■ Anaerobic Chemotrophs such as formate; some can use methanol or acetate as well.
Representative genera of methanogens include Methanospirillum
Learning Outcome and Methanosarcina (figure 11.1).
1. Compare and contrast the characteristics and habitats of Methanogens are found in anaerobic environments where H2 and
methanogens, sulfur- and sulfate-reducing bacteria, Clostridium CO2 are both available. Because these gases are generated by bacteria
species, lactic acid bacteria, and Propionibacterium species. that ferment organic material, methanogens often grow in association
with these microbes. Methanogens, however, are generally not found
For approximately the first 1.5 billion years that prokaryotes in environments containing high levels of sulfate, nitrate, or other
inhabited earth, the atmosphere was anoxic, or devoid of O2. In inorganic electron acceptors. This is because microorganisms that
that anaerobic environment, some early chemotrophs (organ- use these electron acceptors to oxidize H2 have a competitive advan-
isms that harvest energy by oxidizing chemicals) probably used tage; the use of CO2 as an electron acceptor releases comparatively
pathways of anaerobic respiration, employing terminal electron little energy (see figure  6.7). Environments where methanogens are
acceptors such as carbon dioxide or elemental sulfur, which were commonly found include sewage, swamps, marine sediments, rice
plentiful in the environment. Others may have used fermentation, paddies, and the digestive tracts of humans and other animals. The
passing the electrons to an organic molecule such as pyruvate. methane produced can be seen as bubbles rising in swamp waters or
chemotrophs, p.  93 anaerobic respiration, pp.  134, 144 terminal the 10 cubic feet of gas discharged from a cow’s digestive system each
electron acceptor, p. 130 fermentation, pp. 134, 147 day. As a by-product of sewage treatment plants, methane gas can be
Today, anaerobic habitats are still common. Mud and tightly collected and used for heating and generating electricity.
packed soil limit the diffusion of gases, and any O2 that penetrates is Studying methanogens is challenging because they are very
rapidly consumed by aerobically respiring organisms. This creates sensitive to O2. Special techniques, including anaerobe chambers,
anaerobic conditions just below the surface. Aquatic environments are used to culture them. culturing anaerobes, p. 96
may also become anaerobic if they contain nutrients that foster the
rapid growth of O2-consuming microbes. This is evident in polluted
lakes, where fish may die because of a lack of dissolved O2. The
human body also provides many anaerobic environments, such as
in the intestinal tract. Even the skin and the oral cavity, which are
routinely exposed to O2, have anaerobic microenvironments. These
are created via the localized depletion of O2 by aerobes.

MicroByte
Approximately 99% of the prokaryotes that inhabit the intestinal
tract are obligate anaerobes.

Anaerobic Chemolithotrophs
Chemolithotrophs oxidize reduced inorganic chemicals such
as hydrogen gas (H2) to obtain energy. Those growing anaerobi-
cally obviously cannot use O2 as a terminal electron acceptor
and instead must employ an alternative such as carbon dioxide
or sulfur. Relatively few anaerobic chemolithotrophs have been
discovered, and most are members of the domain Archaea. Some
bacterial examples that inhabit aquatic environments will be dis- (a) 5 μm (b) 5 μm
cussed later. chemolithotrophs, p. 93
FIGURE 11.1 Methanogens (a) Phase-contrast micrograph of
a Methanospirillum species. (b) Scanning electron micrograph of a
The Methanogens Methanosarcina species.
Methanogens are a group of archaea that generate ATP by oxi- ? What roles do hydrogen gas and carbon dioxide play in the
dizing hydrogen gas, using CO2 as a terminal electron acceptor. metabolism of methanogens?
 Part II The Microbial World 257

TABLE 11.1 Metabolic Diversity of Prokaryotes

Group/Genera Characteristics Phylum

Anaerobic Chemolithotrophs
Methanogens—Methanospirillum, Members of the Archaea that oxidize hydrogen gas, using CO2 as a terminal Euryarchaeota
Methanosarcina electron acceptor to generate methane.
Anaerobic Chemoorganotrophs—Anaerobic Respiration
Sulfur- and sulfate-reducing Use sulfate as a terminal electron acceptor, generating hydrogen sulfide. Proteobacteria
bacteria—Desulfovibrio Found in anaerobic muds rich in organic material. Gram-negative.
Anaerobic Chemoorganotrophs—Fermentation
Clostridium Endospore-forming obligate anaerobes. Inhabitants of soil. Gram-positive. Firmicutes
Lactic acid bacteria—Streptococcus, Produce lactic acid as the major end product of their fermentative Firmicutes
Enterococcus, Lactococcus, metabolism. Aerotolerant anaerobes. Several genera are used by the food
Lactobacillus, Leuconostoc industry. Gram-positive.
Propionibacterium Obligate anaerobes that produce propionic acid as their primary Actinobacteria
fermentation end product. Used in the production of Swiss cheese.
Gram-positive.
Anoxygenic Phototrophs
Purple sulfur bacteria—Chromatium, Grow in colored masses in sulfur-rich aquatic habitats, using sulfur Proteobacteria
Thiospirillum, Thiodictyon compounds as a source of electrons when making reducing power.
Gram-negative.
Purple non-sulfur bacteria— Grow in aquatic habitats, preferentially using organic compounds as a Proteobacteria
Rhodobacter, Rhodopseudomonas source of electrons for reducing power. Many are metabolically versatile.
Gram-negative.
Green sulfur bacteria—Chlorobium, Found in habitats similar to those preferred by the purple sulfur bacteria. Chlorobi
Pelodictyon Gram-negative.
Filamentous anoxygenic Characterized by their filamentous growth. Gram-negative. Chloroflexi
phototrophic bacteria—Chloroflexus
Others—Heliobacterium Have not been studied extensively. Firmicutes
Oxygenic Phototrophs—Cyanobacteria
Anabaena, Synechococcus Important primary producers. Some fix N2. Gram-negative. Cyanobacteria

Aerobic Chemolithotrophs
Filamentous sulfur oxidizers— Oxidize sulfur compounds as energy sources. Found in sulfur springs and Proteobacteria
Beggiatoa, Thiothrix sewage-polluted waters. Gram-negative.
Unicellular sulfur oxidizers— Oxidize sulfur compounds as energy sources. Some species produce enough Proteobacteria
Thiobacillus, Acidithiobacillus acid to lower the pH to 1.0. Gram-negative.
Nitrifiers—Nitrosomonas, Oxidize ammonia or nitrite as energy sources. This converts certain Proteobacteria
Nitrosococcus, Nitrobacter, fertilizers to a form easily leached from soils, and depletes O2 in waters
Nitrococcus polluted with ammonia-containing wastes. Genera that oxidize nitrite
prevent the toxic buildup of nitrite. Gram-negative.
Hydrogen-oxidizing bacteria— Thermophilic bacteria that oxidize hydrogen gas as an energy source. One Aquifacae
Aquifex, Hydrogenobacter of the earliest bacterial forms to exist on earth.
Aerobic Chemoorganotrophs—Obligate Aerobes
Micrococcus Widely distributed; common laboratory contaminants. Gram-positive. Actinobacteria
Mycobacterium Waxy cell wall resists staining; acid-fast. Actinobacteria
Pseudomonas Common environmental bacteria that, as a group, can degrade a wide Proteobacteria
variety of compounds. Gram-negative.
Thermus Thermus aquaticus is the source of Taq polymerase (used in PCR). Stains Deinococcus-
Gram-negative. Thermus
Deinococcus Resistant to the damaging effects of gamma radiation. Stains Gram-positive. Deinococcus-
Thermus
Aerobic Chemoorganotrophs—Facultative Anaerobes
Corynebacterium Widespread in nature. Gram-positive. Actinobacteria
The Enterobacteriaceae—Escherichia, Most reside in the intestinal tract. Those that ferment lactose are coliforms; Proteobacteria
Enterobacter, Klebsiella, Proteus, their presence in water serves as an indicator of fecal pollution. Gram-
Salmonella, Shigella, Yersinia negative.
258 Chapter 11 The Diversity of Prokaryotic Organisms

Anaerobic Chemoorganotrophs— bacteria multiply. Vegetative cells that develop from endospores
are responsible for a variety of diseases, including tetanus (caused
Anaerobic Respiration
by C. tetani), gas gangrene (caused by C. perfringens), and botu-
Chemoorganotrophs oxidize organic compounds such as glucose lism (caused by C. botulinum). Some species of Clostridium are
to obtain energy. Those that grow anaerobically often use sulfur normal inhabitants of the intestinal tract of humans and other
or sulfate as a terminal electron acceptor. chemoorganotrophs, p. 93 animals. endospores, p.  67 tetanus, p.  555 gas gangrene, p.  558
botulism, p. 652
Sulfur- and Sulfate-Reducing Bacteria As a group, Clostridium species ferment a wide variety of
When sulfur compounds are used as terminal electron acceptors, compounds, including sugars and cellulose. Some of the end prod-
they become reduced to form hydrogen sulfide, the compound ucts are commercially valuable—for example, C. acetobutylicum
responsible for the rotten-egg smell of many anaerobic environ- produces acetone and butanol. Some species can ferment amino
ments. An example of this reaction is: acids by an unusual process that oxidizes one amino acid, using
another as a terminal electron acceptor. This generates a variety of
organic compounds + S → CO2 + H2S
foul-smelling end products associated with rotting flesh.
(energy source) (terminal electron
acceptor)
The Lactic Acid Bacteria
In addition to its unpleasant odor, H2S is a problem to industry Gram-positive bacteria that produce lactic acid as a major end
because it reacts with metals, corroding pipes and other structures. product of their fermentative metabolism make up a group called
Ecologically, however, prokaryotes that reduce sulfur compounds the lactic acid bacteria. This includes members of the genera
are an essential component of the sulfur cycle. sulfur cycle, p. 728 Streptococcus, Enterococcus, Lactococcus, Lactobacillus, and
Sulfate- and sulfur-reducing bacteria generally live in Leuconostoc. Most can grow in aerobic environments, but they
mud that has organic material and oxidized sulfur compounds. only ferment. They can be easily distinguished from other bacteria
The H2S they produce causes mud and water to turn black that grow in the presence of O2 because they lack the enzyme
when it reacts with iron molecules. At least a dozen genera catalase (see figure 10.4a). obligate fermenters, p. 90 catalase, p. 90
are recognized in this group, the most extensively studied of Streptococcus species are cocci that typically grow in chains
which are species of Desulfovibrio. These are Gram-negative of varying lengths (figure 11.2). They inhabit the oral cavity, gen-
curved rods. erally as part of the normal microbiota. Some, however, are patho-
Some representatives of the Archaea also use sulfur com- gens. One of the most important is S. pyogenes (group A strep),
pounds as terminal electron acceptors, but the characterized which causes pharyngitis (strep throat) and other diseases. Unlike
examples generally do not inhabit the same environments as the streptococci that typically inhabit the throat, S. pyogenes is
their bacterial counterparts. Although most of the sulfur-reducing β-hemolytic, an important characteristic used to distinguish it
bacteria are either mesophiles or thermophiles, the known sulfur- from most members of the normal microbiota (see figure  4.10).
reducing archaea are hyperthermophiles, inhabiting extreme envi-  Streptococcus pyogenes, p. 487 hemolysis, p. 96
ronments such as hydrothermal vents. They will be discussed later Species of Lactococcus and Enterococcus were at one time
in the chapter. thermophiles, p. 90 hyperthermophiles, p. 90 included in the genus Streptococcus. The genus Lactococcus now
includes species used by the dairy industry to produce fermented
Anaerobic Chemoorganotrophs— milk products such as cheese and yogurt. Enterococcus species
typically inhabit the intestinal tract of humans and other animals.
Fermentation fermentation of dairy products, p. 751
Numerous types of anaerobic bacteria obtain energy by ferment- Members of the genus Lactobacillus are rod-shaped bacteria
ing, producing ATP only by substrate-level phosphorylation. that grow as single cells or loosely associated chains. They are
There are many variations of fermentation, using different organic common members of the microbiota in the mouth and the healthy
energy sources and producing characteristic end products, but one vagina during childbearing years. In the vagina, they break down
example is: glycogen that has been deposited in the vaginal lining in response
glucose → pyruvate → lactic acid to estrogen (the female sex hormone). The resulting low pH helps
(energy source) (terminal electron prevent vaginal infections. Lactobacilli are also often present in
acceptor) decomposing plant material, milk, and other dairy products. Like
the lactococci, they are important in the production of fermented
The Genus Clostridium foods (figure 11.3).
Members of the genus Clostridium are Gram-positive rods that
can form endospores (see figure  3.45). They are common soil The Genus Propionibacterium
inhabitants, and the vegetative cells live in the anaerobic micro- Propionibacterium species are Gram-positive pleomorphic (irregular-
environments created when aerobic organisms consume available shaped) rods that produce propionic acid as their primary fer-
O2. Their endospores can tolerate O2 and survive for long periods mentation end product. They can also ferment lactic acid, thereby
by withstanding levels of heat, drying, chemicals, and irradia- extracting energy from a waste product of other bacteria.
tion that would kill vegetative bacteria. When conditions become Propionibacterium species are valuable to the dairy industry
favorable, these endospores germinate, and the resulting vegetative because their fermentation end products are important in Swiss
 Part II The Microbial World 259

FIGURE 11.2
Streptococcus Species
(a) Gram stain. (b) Scanning
electron micrograph.
? What is the major
metabolic end product of
Streptococcus species?

(a) 10 μm (b) 1 μm

cheese production. The propionic acid gives the typical nutty 11.2 ■ Anoxygenic Phototrophs
flavor of the cheese, and CO2, also a product of the fermentation,
creates the signature holes. Propionibacterium species are also Learning Outcome
found in the intestinal tract and in anaerobic microenvironments 2. Compare and contrast the characteristics of the purple bacteria
on the skin. Swiss cheese production, p. 751 and the green bacteria.

MicroAssessment 11.1 The earliest photosynthesizing organisms were likely anoxygenic

Methanogens are archaea that oxidize hydrogen gas, using CO2 as
phototrophs. Rather than using water as a source of electrons
a terminal electron acceptor, to generate methane. The sulfur- and when making reducing power for biosynthesis, these organisms
sulfate-reducing bacteria oxidize organic compounds, with sulfur use hydrogen sulfide or organic compounds, and therefore do not
or sulfate serving as a terminal electron acceptor, to generate generate O2. For example:
hydrogen sulfide. Clostridium species, the lactic acid bacteria, and
Propionibacterium species oxidize organic compounds, with an 6 CO2 + 12 H2S → C6H12O6 + 12 S + 6 H2O
organic compound serving as a terminal electron acceptor. (carbon (electron
1. What metabolic process creates the rotten-egg smell source) source)
characteristic of many anaerobic environments? Modern-day anoxygenic phototrophs are a phylogenetically diverse
2. Describe two beneficial contributions of the lactic acid bacteria. group of bacteria that live in environments that have little or no O2
3. Relatively little is known about many obligate anaerobes. Why yet light penetrates. Typical habitats include bogs, lakes, and the
would this be so? + upper layer of muds. reducing power, p. 131
As discussed in chapter 6, the photosystems of the anoxy-
genic phototrophs are different from those of plants, algae, and
cyanobacteria. They have a unique type of chlorophyll called bac-
teriochlorophyll. This and their other light-harvesting pigments
absorb wavelengths that penetrate deeper than those absorbed by
chlorophyll a. photosystems, p. 151 chlorophyll a, p. 152

The Purple Bacteria

The purple bacteria are Gram-negative organisms that appear
red, orange, or purple due to their light-harvesting pigments.
Unlike other anoxygenic phototrophs, the components of their
photosynthetic apparatus are all contained within the cytoplasmic
membrane. Folds in this membrane increase the surface area avail-
able for the photosynthetic processes.

Purple Sulfur Bacteria

Purple sulfur bacteria can sometimes be seen growing as col-
5 μm
ored masses in sulfur-rich aquatic habitats such as sulfur springs
FIGURE 11.3 Lactobacillus Species from Yogurt (figure  11.4a). The cells are relatively large, sometimes larger
? Lactobacillus species are common members of the normal than 5 μm in diameter, and some are motile by flagella. They may
microbiota of which human body sites? also have gas vesicles, allowing them to move up or down to their
260 Chapter 11 The Diversity of Prokaryotic Organisms

Sulfur granules

(a) (b) 10 μm

FIGURE 11.4 Purple Sulfur Bacteria (a) Photograph of bacteria growing in a bog. (b) Photomicrograph showing intracellular sulfur granules.
? What is the role of the sulfur granules in the bacterial cells?

preferred level in the water column. Most store sulfur in intracel- of the cell (figure 11.5). The accessory pigments of the green sulfur
lular granules (figure 11.4b). gas vesicles, p. 67 bacteria are located in structures called chlorosomes. The bacteria
The purple sulfur bacteria preferentially use hydrogen sulfide lack flagella, but many have gas vesicles. All are strict anaerobes,
to generate reducing power, although some can use other inorganic and none can use a chemotrophic metabolism. Representative gen-
molecules (such as H2) or organic compounds (such as pyruvate). era include Chlorobium and Pelodictyon. accessory pigments, p. 152
Many are strict anaerobes and phototrophs, but some can grow in
the absence of light aerobically, oxidizing reduced inorganic or Filamentous Anoxygenic Phototrophic Bacteria
organic compounds as a source of energy. Representative genera Filamentous anoxygenic phototrophic bacteria form multicellular
include Chromatium, Thiospirillum, and Thiodictyon. arrangements and exhibit gliding motility. The most thoroughly
studied of this group are members of the genus Chloroflexus, par-
Purple Non-Sulfur Bacteria ticularly the thermophilic strains that grow in hot springs. Many of
The purple non-sulfur bacteria are found in a wide variety of the filamentous anoxygenic phototrophs have chlorosomes, which
aquatic habitats, including moist soils, bogs, and paddy fields. initially led scientists to believe they were related to the green
One important characteristic that distinguishes them from the sulfur bacteria. Their 16S rDNA sequences indicate otherwise. As
purple sulfur bacteria is that they preferentially use a variety of a group, filamentous anoxygenic phototrophs are diverse meta-
organic molecules rather than hydrogen sulfide as a source of bolically. Some preferentially use organic compounds to generate
electrons for reducing power. In addition, they lack gas vesicles. reducing power and can also grow in the dark aerobically using
If they do store sulfur, the granules form outside the cell. chemotrophic metabolism.
Purple non-sulfur bacteria are remarkably versatile metaboli-
cally. Not only do they grow as phototrophs using organic mol-
ecules as a source of electrons, but many can use a metabolism
similar to the purple sulfur bacteria, employing hydrogen gas or
hydrogen sulfide as an electron source. In addition, most can grow
aerobically in the absence of light using chemotrophic metabo-
lism. Representative genera of purple non-sulfur bacteria include
Rhodobacter and Rhodopseudomonas.

The Green Bacteria Sulfur granules

The green bacteria are Gram-negative organisms that are typi-
cally green or brownish in color.

Green Sulfur Bacteria

Green sulfur bacteria are found in habitats similar to those preferred 5 μm

by the purple sulfur bacteria. Like the purple sulfur bacteria, they FIGURE 11.5 Green Sulfur Bacteria Note that the sulfur
use hydrogen sulfide as a source of electrons for reducing power granules are extracellular.
and they form sulfur granules. The granules, however, form outside ? What role does sulfur play in the metabolism of green sulfur bacteria?
 Part II The Microbial World 261

Other Anoxygenic Phototrophs

Although the green and purple bacteria have been studied most exten-
sively, other types of anoxygenic phototrophs exist. Among these are
members of the genus Heliobacterium, Gram-positive endospore-
forming rods related to members of the genus Clostridium.

MicroAssessment 11.2
Anoxygenic phototrophs harvest the energy of sunlight, but do not
generate O2. The purple sulfur bacteria and green sulfur bacteria
use hydrogen sulfide as a source of electrons to generate reducing (a) 15 μm
power; the purple non-sulfur bacteria and many of the filamentous
anoxygenic phototrophs preferentially use organic compounds.
4. Describe a structural characteristic that distinguishes the purple
sulfur bacteria from the green sulfur bacteria.
5. What is the function of gas vesicles?
6. How do anoxygenic phototrophs benefit by having light-
harvesting pigments that absorb wavelengths that penetrate
deeper than those absorbed by chlorophyll a? +

11.3 ■ Oxygenic Phototrophs (b) 100 μm

FIGURE 11.6 Cyanobacteria (a) The spiral trichome of Spirulina
Learning Outcome species. (b) Differential interference contrast photomicrograph of a
3. Describe the characteristics of cyanobacteria, including how species of Oscillatoria. Note the arrangement of the individual cells in
nitrogen-fixing species protect their nitrogenase enzyme from O2. the trichome.
? What is a trichome?
Nearly 3 billion years ago, the earth’s atmosphere began changing
as O2 was gradually introduced to the previously anoxic environ-
ment. This was probably due to the evolution of the cyanobacteria,
thought to be the earliest oxygenic phototrophs. These photosyn- that holds and surrounds a chain of cells) (figure  11.6). Motile
thetic organisms use water as a source of electrons for reducing trichomes glide as a unit. Cyanobacteria that inhabit aquatic
power, generating O2: environments often have gas vesicles, allowing them to move
vertically within the water column. When large numbers of cyano-
6 CO2 + 6 H2O → C6H12O6 + 6 O2 bacteria accumulate in stagnant lakes or other freshwater habitats,
(carbon source) (electron source) it is called a bloom (figure 11.7). In the bright, hot conditions of
Cyanobacteria still play an essential role in the biosphere. As primary
producers, they harvest the energy of sunlight, using it to convert
CO2 into organic compounds. They were initially thought to be
algae and were called blue-green algae until electron microscopy
revealed their prokaryotic structure. primary producers, p. 719

The Cyanobacteria
Cyanobacteria are a diverse group of more than 60 genera of
Gram-negative bacteria. They inhabit a wide range of environments,
including freshwater and marine habitats, soils, and the surfaces of
rocks. In addition to being photosynthetic, many are able to convert
nitrogen gas (N2) to ammonia, which can then be incorporated into
cell material. This process, called nitrogen fixation, is an exclusive
ability of prokaryotes. nitrogen fixation, p. 726

General Characteristics of Cyanobacteria

Cyanobacteria are morphologically diverse. Some are unicellular, FIGURE 11.7 Cyanobacterial Bloom Excessive growth of cyano-
with typical prokaryotic shapes such as cocci, rods, and spirals. bacteria is evidenced by buoyant masses of cells that have risen to the
Others form filamentous multicellular associations called tri- surface.
chomes that may or may not be enclosed within a sheath (a tube ? What allows cyanobacteria cells to rise to the surface?
262 Chapter 11 The Diversity of Prokaryotic Organisms

summer, the buoyant cells lyse and decay, creating a foul-smelling nitrogen. Synechococcus species fix nitrogen only in the dark.
scum. The ecological effects of these blooms on aquatic habitats Consequently, nitrogen fixation and photosynthesis are tempo-
are discussed in chapter 29. aquatic habitats, p. 722 rally separated. photosystem II, p. 153
The photosystems of the cyanobacteria are like those con-
tained within the chloroplasts of algae and plants. This is not Other Notable Characteristics
surprising in light of the genetic evidence indicating chloroplasts of Cyanobacteria
evolved from a species of cyanobacteria that once resided as an Cyanobacteria have various other notable characteristics—some
endosymbiont within eukaryotic cells (see Perspective 3.1). In beneficial, others damaging. Filamentous cyanobacteria are
addition to light-harvesting chlorophyll pigments, cyanobacte- responsible for maintaining the structure and productivity of soils
ria have phycobiliproteins. These pigments absorb energy from in cold desert areas such as the Colorado Plateau. Their sheaths
wavelengths of light not well absorbed by chlorophyll. They persist in soil, creating a sticky fibrous network that prevents
contribute to the blue-green, or sometimes reddish, color of the erosion. In addition, these bacteria provide an important source
cyanobacteria. photosystem, p. 151 of nitrogen and organic carbon in otherwise nutrient-poor soils.
On the negative side, some cyanobacteria produce geosmin, a
Nitrogen-Fixing Cyanobacteria chemical that has a distinctive “earthy” odor, which makes drink-
Nitrogen-fixing cyanobacteria are critical ecologically. They can ing water taste odd. Some aquatic species such as Microcystis
incorporate both N2 and CO2 into organic material, so they gener- aeruginosa produce toxins that can be deadly to an animal when
ate a form of these nutrients that can then be used by other organ- consumed.
isms. Thus, their activities can ultimately support the growth of
a wide range of organisms in environments that would otherwise MicroAssessment 11.3
lack usable nitrogen and carbon. As an example, nitrogen-fixing
The photosystems of cyanobacteria generate O2 and are similar to
cyanobacteria in the oceans are essential primary producers that those of algae and plants. Many cyanobacteria can fix nitrogen.
support other sea life. Also, like all cyanobacteria, they help limit
7. What is the function of a heterocyst?
atmospheric CO2 buildup by using the gas as a carbon source.
Nitrogenase, the enzyme complex that catalyzes nitrogen 8. How do cyanobacteria prevent erosion in cold desert regions?
fixation, is destroyed by O2; therefore, nitrogen-fixing cyano- 9. How could heavily fertilized lawns foster the development of
bacteria must protect the enzyme from the O2 they generate. cyanobacterial blooms? +
Species of Anabaena, which are filamentous, isolate nitroge-
nase by confining the process of nitrogen fixation to specialized
thick-walled cells called heterocysts (figure  11.8). Heterocysts
lack photosystem II and consequently do not generate O2. The
11.4 ■ Aerobic Chemolithotrophs
heterocysts of some species form at very regular intervals within Learning Outcome
the filament, reflecting the ability of cells within a trichome to
4. Compare and contrast the characteristics of sulfur-oxidizing
communicate. One species of Anabaena, A. azollae, forms an
bacteria, nitrifiers, and hydrogen-oxidizing bacteria.
intimate relationship with the water fern Azolla. The bacterium
grows and fixes nitrogen within the protected environment of a Aerobic chemolithotrophs obtain energy by oxidizing reduced
special sac in the fern, providing Azolla with a source of available inorganic chemicals, using O2 as a terminal electron acceptor.

The Sulfur-Oxidizing Bacteria

The sulfur-oxidizing bacteria are Gram-negative rods or spirals,
which sometimes grow in filaments. They obtain energy by oxi-
dizing elemental sulfur and reduced sulfur compounds, including
hydrogen sulfide and thiosulfate. O2 serves as a terminal electron
acceptor, generating sulfuric acid. An example of this reaction is:
Heterocyst S + 1* O2 + H2O → H2SO4
(energy source) (terminal electron
acceptor)
These bacteria are important in the sulfur cycle. sulfur cycle, p. 728

10 μm Filamentous Sulfur Oxidizers

FIGURE 11.8 Heterocyst of an Anabaena Species Nitrogen Beggiatoa and Thiothrix species are filamentous sulfur oxidiz-
fixation occurs within these specialized cells. ers that live in sulfur springs, in sewage-polluted waters, and
? Why is it important for heterocysts to not have a functional on the surface of marine and freshwater sediments. They store
photosystem II? sulfur, depositing it as intracellular granules, but differ in the
 Part II The Microbial World 263

Perspective  6.1). The bacteria oxidize insoluble metal sulfides

such as gold sulfide, producing sulfuric acid. This lowers the pH,
which converts the metal to a soluble form. Acidithiobacillus spe-
Multicellular cies can also be used to prevent acid rain, a problem that occurs
filament
when sulfur-containing coals and oils are burned. To remove the
sulfur from the fuels, the bacteria are allowed to oxidize it to sul-
fate, a form that can then be extracted.
Sulfur
Acidithiobacillus species can also cause severe environmental
granules problems. For example, the strip mining of coal exposes metal
sulfides, which the bacteria can then oxidize to produce sulfuric
acid; some species produce enough acid to lower the pH to 1.0.
Cellular The resulting runoff can acidify nearby streams, killing trees, fish,
septa
and other wildlife (figure  11.10). The runoff may also contain
toxic metals made soluble by the bacteria.
(a) 10 μm

The Nitrifiers
Nitrifiers are a diverse group of Gram-negative bacteria that
obtain energy by oxidizing inorganic nitrogen compounds such as
ammonia or nitrite. These bacteria are a concern to farmers who
fertilize their crops with ammonium nitrogen, a form of nitrogen
retained by soils because its positive charge causes it to adhere
to negatively charged soil particles. The potency and longevity
of the fertilizer are affected by nitrifying bacteria converting the
ammonia to nitrate. Although plants use the nitrate more easily, it
is rapidly leached from soils.
Nitrifying bacteria are also an important consideration when
disposing of wastes that have a high ammonia concentration. As
nitrifying bacteria oxidize nitrogen compounds, they consume O2,
(b) 10 μm so waters polluted with nitrogen-containing wastes can quickly
FIGURE 11.9 Filamentous Sulfur-Oxidizing Bacteria Phase- become hypoxic (low in dissolved O2).
contrast photomicrographs. (a) Multicellular filament of a Beggiatoa The nitrifiers include two metabolically distinct groups that
species; the septa separate the cells. (b) Multicellular filaments of a
typically grow in close association. Together, they can oxidize
Thiothrix species, forming a rosette arrangement.
? What is the role of sulfur in the metabolism of sulfur-oxidizing
bacteria?

FIGURE 11.10 Acid Drainage from a Mine Sulfur-oxidizing

nature of their filamentous growth (figure  11.9). The filaments bacteria oxidize exposed metal sulfides, generating sulfuric acid. The
yellow-red color is due to insoluble iron oxides.
of Beggiatoa species move by gliding motility, a mechanism that
does not require flagella. The filaments may flex or twist to form ? How can the metabolic activities that result in this acid drainage
a tuft. In contrast, the filaments of Thiothrix species are immobile; be used in a commercially valuable manner?
they fasten at one end to rocks or other solid surfaces. Often they
attach to other cells, forming characteristic rosette arrangements
of filaments. Progeny cells detach from the ends of these fila-
ments and use gliding motility to move to new locations, where
they form additional filaments. Overgrowth of these filamentous
organisms in wastewater at treatment facilities causes a problem
called bulking. Because the masses of filamentous organisms do
not settle easily, bulking interferes with the separation of the solid
sludge from the liquid effluent. wastewater treatment, p. 736

Unicellular Sulfur Oxidizers

Acidithiobacillus species are found in both terrestrial and aquatic
habitats, where their ability to oxidize metal sulfides can be
used for bioleaching, a process used to recover metals (see
264 Chapter 11 The Diversity of Prokaryotic Organisms

ammonia to form nitrate. The ammonia oxidizers, which include Aerobic chemoorganotrophs oxidize organic compounds to obtain
the genera Nitrosomonas and Nitrosococcus, convert ammonia to energy, using O2 as a terminal electron acceptor:
nitrite in the following reaction:
organic compounds + O2 → CO2 + H2O
NH4+ + 1* O2 → NO2– + H2O + 2 H+ (energy source) (terminal electron
(energy source) (terminal electron acceptor)
acceptor) They include a wide assortment of bacteria, ranging from some
that inhabit very specific environments to others that are ubiqui-
The nitrite oxidizers, which include the genera Nitrobacter and
tous. This section will profile only representative genera found in
Nitrococcus, then convert nitrite to nitrate as follows:
a variety of different environments. Later sections will describe
NO2– + * O2 → NO3– examples that thrive in specific habitats.
(energy source) (terminal electron
acceptor) Obligate Aerobes
The latter group is particularly important in preventing the buildup Obligate aerobes obtain energy using respiration exclusively;
of nitrite in soils, which is toxic and can leach into groundwater. none can ferment.
The oxidation of ammonia to nitrate (nitrification) is an important The Genus Micrococcus
part of the nitrogen cycle. nitrogen cycle, p. 726
Members of the genus Micrococcus are Gram-positive cocci found
in soil and on dust particles, inanimate objects, and skin. Because
they are often airborne, they can easily contaminate bacteriological
The Hydrogen-Oxidizing Bacteria media. There, they typically form pigmented colonies, a character-
Members of the Gram-negative genera Aquifex and istic that helps identify them. The colonies of M. luteus, for exam-
Hydrogenobacter are among the few hydrogen-oxidizing bacteria ple, are generally yellow (figure 11.11). Like members of the genus
that are obligate chemolithotrophs. An example of the reaction in Staphylococcus, which will be discussed later, they tolerate dry
their metabolism is: conditions and can grow in salty environments such as 7.5% NaCl.

H2 + * O2 → H2O The Genus Mycobacterium

(energy source) (terminal electron Mycobacterium species are widespread in nature and include harm-
acceptor) less saprophytes, which live on dead and decaying matter, as well
as pathogens. They stain poorly because of a waxy lipid (mycolic
These related organisms are thermophilic and typically inhabit hot
acid) in their unusual cell wall, but special procedures can be used
springs. Some Aquifex species have a maximum growth tempera-
to increase the penetration of certain dyes. Once stained, the cells
ture of 95°C, the highest of any bacteria. The hydrogen-oxidizing
resist destaining, even with acidic decolorizing solutions. Because
bacteria are deeply branching in the phylogenetic tree, meaning
of this, Mycobacterium species are called acid-fast, and the acid-
that according to 16S rRNA studies, they were one of the earli-
fast staining procedure is an important step in identifying them
est bacterial forms to exist on earth. The fact that they require O2
(see figure  3.15). Nocardia species, a related group of bacteria
seems contradictory to their evolutionary position, but in fact,
common in soil, are also acid-fast. acid-fast, p. 48
the low amount they require might have been available early on
in certain niches due to photochemical processes that split water.

MicroAssessment 11.4
Sulfur oxidizers use sulfur compounds as energy sources, generating
sulfuric acid. The nitrifiers oxidize nitrogen compounds such as
ammonium or nitrite. Hydrogen-oxidizing bacteria oxidize H2.
10. What is the role of sulfur oxidizers in bioleaching?
11. Why would farmers be concerned about nitrifying bacteria?
12. Why would sulfur-oxidizing bacteria store sulfur? +

11.5 ■ Aerobic Chemoorganotrophs

Learning Outcomes
5. Describe the representative obligate aerobes and facultative
anaerobes.
FIGURE 11.11 Micrococcus Iuteus Colonies
6. Describe the family Enterobacteriaceae, and explain what
distinguishes coliforms from other members of this family. ? Why is Micrococcus luteus a common contaminate on
bacteriological media?
 Part II The Microbial World 265

Mycobacterium species are generally pleomorphic rods; they species are thermophilic, as their name implies, and this trait is
often occur in chains that sometimes branch, or bunch together to valuable because of their heat-stable enzymes. The bacteria have
form cordlike groups. Several species are notable for their effect an unusual cell wall, and stain Gram-negative. PCR, p. 227
on human health, including M. tuberculosis, which causes tuber- Deinococcus species’ unusual cell wall has multiple layers,
culosis, and M. leprae, which causes Hansen’s disease (leprosy). and they stain Gram-positive. They are unique in their extraor-
Mycobacterium species are more resistant to disinfectants than dinary resistance to the damaging effects of gamma radiation.
most other vegetative bacteria. In addition, they are resistant to D.  radiodurans can survive a radiation dose several thousand
many of the most common antimicrobial medications. times that lethal to a human being. The dose literally shatters the
organism’s genome into many fragments, yet enzymes in the cells
The Genus Pseudomonas can repair the extensive damage. Scientists anticipate that through
Pseudomonas species are Gram-negative rods that have polar flagella genetic engineering, Deinococcus species may eventually help
and often produce pigments (figure 11.12). Although most are strict clean up the soil and water contaminated by the radioactive wastes
aerobes, some can grow anaerobically if nitrate is available as a termi- that have accumulated in the United States.
nal electron acceptor. They do not ferment and are oxidase-positive,
MicroByte
characteristics that help distinguish them from members of the family
The DNA polymerase of T. aquaticus (Taq polymerase) is a
Enterobacteriaceae, including E. coli. oxidase test, p. 243 fundamental part of the polymerase chain reaction (PCR).
As a group, Pseudomonas species have extremely diverse bio-
chemical capabilities. Some can metabolize more than 80 different
substrates, including unusual sugars, amino acids, and compounds Facultative Anaerobes
containing aromatic rings. Because of this, Pseudomonas species play Facultative anaerobes preferentially use aerobic respiration if O2
an important role in the degradation of many synthetic and natural is available. As an alternative, however, they can ferment.
compounds that resist breakdown by most other microorganisms. The
ability to carry out some of these degradations is encoded by plasmids. The Genus Corynebacterium
Pseudomonas species are widespread, typically inhabiting Members of the genus Corynebacterium are widespread in nature.
soil and water. Although most are harmless, some cause disease They are Gram-positive pleomorphic rods, often club-shaped and
in plants and animals. Medically, the most significant species is arranged to form V shapes or palisades (koryne is Greek for “club”)
P.  aeruginosa. It is a common opportunistic pathogen, meaning (figure 11.13). Bacteria that exhibit this characteristic morphology
that it primarily infects people who have underlying medical are referred to as coryneforms or diphtheroids. Corynebacterium
conditions. Unfortunately, it can grow in nutrient-poor environ- species are generally facultative anaerobes, but some are strict
ments, such as water used in respirators, and is resistant to many aerobes. Many Corynebacterium species reside harmlessly in the
disinfectants and antimicrobial medications. Because of this, hos- throat, but toxin-producing strains of C. diphtheriae can cause the
pitals must be very careful to prevent it from infecting patients. disease diphtheria. diphtheria, p. 490
 Pseudomonas aeruginosa, p. 554 opportunistic pathogen, p. 382
The Family Enterobacteriaceae
The Genera Thermus and Deinococcus Members of the family Enterobacteriaceae, often referred to
Thermus and Deinococcus are related genera that have scien- as enterics or enterobacteria, are Gram-negative rods. Their
tifically and commercially important characteristics. Thermus name reflects the fact that most reside in the intestinal tract of

FIGURE 11.12 Pigments of Pseudomonas Species Cultures of

different strains of Pseudomonas aeruginosa. Note the different col- 10 μm
ors of the water-soluble pigments. FIGURE 11.13 Corynebacterium The Gram-positive pleomorphic
rods are often arranged to form V shapes or palisades.
? Why is the fact that Pseudomonas species can grow in nutrient-
poor environments medically important? ? How does the name of this genus reflect the shape of the bacteria?
266 Chapter 11 The Diversity of Prokaryotic Organisms

humans and other animals (in Greek enteron means “intestine”), of certain common intestinal inhabitants such as E. coli that are
although some thrive in rich soil. Enterics that are part of the nor- easy to detect in food and water; for years regulatory agencies have
mal intestinal microbiota include Enterobacter, Klebsiella, and considered them to be an indicator of fecal pollution. Their pres-
Proteus species as well as most strains of E. coli. Those that cause ence indicates a possible health risk because fecal-borne pathogens
diarrheal disease include Shigella species, Salmonella enterica, might also be present. coliform, p. 742
and some strains of E. coli. Life-threatening systemic diseases
include typhoid fever, caused by Salmonella enterica serotype
MicroAssessment 11.5
Typhi, and both the bubonic and pneumonic forms of plague,
caused by Yersinia pestis. diarrheal disease, p. 585 typhoid fever, Micrococcus, Mycobacterium, Pseudomonas, Thermus, and
p. 592 plague, p. 678
Deinococcus species are obligate aerobes that harvest energy by
degrading organic compounds, using O2 as a terminal electron
Members of the Enterobacteriaceae are facultative anaerobes acceptor. Most Corynebacterium species and all members of the
that ferment glucose and, if motile, generally have peritrichous family Enterobacteriaceae are facultative anaerobes.
flagella. The family includes over 40 recognized genera that can
13. What unique characteristic makes members of the genus
be distinguished using biochemical tests. Within a given species, Deinococcus noteworthy?
many different strains have been described. These are often distin-
14. What is the significance of finding coliforms in drinking water?
guished using serological tests that detect differences in cell walls,
15. Why would it be an advantage for a Pseudomonas species to
flagella, and capsules (see figure 10.9). peritrichous flagella, p. 64
encode enzymes for degrading certain compounds on a plasmid
Enteric bacteria that characteristically ferment lactose are rather than the chromosome? +
included in a group called coliforms. This is an informal grouping

ECOPHYSIOLOGICAL DIVERSITY
As a group, prokaryotes show remarkable diversity in their physi- Endospore-Formers
ological adaptations to a wide range of habitats. From the hydro- Bacillus and Clostridium species are the most common Gram-
thermal vents of deep oceans to the frozen expanses of Antarctica, positive rod-shaped bacteria that form endospores; the position
prokaryotes have evolved to thrive in virtually all environments, of the spore in the cell can help in identification (figure 11.14).
including many that plants and animals would find inhospitable.
This section will highlight the physiological mechanisms
prokaryotes use to thrive in terrestrial and aquatic environments;
the study of these adaptations is called ecophysiology. The sec-
tion will also describe some examples of bacteria that use animals
as habitats. Table 11.2 summarizes characteristics of the bacteria
covered in this section.

11.6 ■ Thriving in Terrestrial

Environments
Learning Outcomes
7. Describe the bacterial groups that form resting stages.
8. Compare and contrast Agrobacterium species and rhizobia.

Microorganisms that live in soil must endure a variety of condi-

tions. Daily and seasonally, soil can routinely alternate between
wet and dry as well as warm and cold. Nutrient availability can
also cycle from abundant to sparse. To thrive in this ever-changing
environment, microbes have evolved mechanisms to cope with
adverse conditions and to use plants as sources of nutrients.
(a) 5 μm (b) 5 μm
Bacteria That Form a Resting Stage
FIGURE 11.14 Endospore-Formers (a) Endospores forming in the
Several genera that live in soil can form a resting stage that allows mid-portion of the cells of Bacillus anthracis. (b) Endospores forming at
them to survive the dry periods typical in many soils. Of the vari- the ends of the cells in Clostridium tetani. Both of these species can cause
ous types of dormant cells, endospores are by far the most resis- fatal disease, but many other species of endospore-formers are harmless.
tant to environmental extremes. ? Members of which endospore-forming genus are obligate anaerobes?
 Part II The Microbial World 267

TABLE 11.2 Ecophysiological Diversity

Group/Genera Characteristics Phylum

Thriving in Terrestrial Environments

Endospore-formers—Bacillus, Bacillus species include both obligate aerobes and facultative anaerobes; Firmicutes
Clostridium Clostridium species are obligate anaerobes. Gram-positive.
Azotobacter Form cysts. Notable for their ability to fix nitrogen in aerobic conditions. Proteobacteria
Gram-negative.
Myxobacteria—Chondromyces, Congregate to make fruiting bodies; cells within these differentiate to form Proteobacteria
Myxococcus, Stigmatella dormant microcysts. Gram-negative.
Streptomyces Resemble fungi in their pattern of growth; produce antibiotics. Gram- Actinobacteria
positive.
Agrobacterium Cause plant tumors. Scientists use their Ti plasmid to move genes into plant Proteobacteria
cells. Gram-negative.
Rhizobia—Rhizobium, Fix nitrogen; form a symbiotic relationship with legumes. Gram-negative. Proteobacteria
Sinorhizobium, Bradyrhizobium,
Mesorhizobium, Azorhizobium
Thriving in Aquatic Environments
Sheathed bacteria—Sphaerotilus, Form chains of cells enclosed within a protective sheath. Swarmer cells move Proteobacteria
Leptothrix to new locations. Gram-negative.
Prosthecate bacteria—Caulobacter, Appendages increase their surface area. Gram-negative. Proteobacteria
Hyphomicrobium
Bdellovibrio Predator of other bacteria. Gram-negative. Proteobacteria
Bioluminescent bacteria— Some form symbiotic relationships with specific types of squid and fish. Gram- Proteobacteria
Photobacterium, Vibrio fischeri negative.
Legionella Often reside within protozoa. Gram-negative. Proteobacteria
Epulopiscium Very large cigar-shaped bacteria that multiply by releasing several daughter Firmicutes
cells; each cell has thousands of copies of the genome. Gram-positive.
Free-living spirochetes— Long spiral-shaped bacteria that move by means of endoflagella. Spirochaetes
Spirochaeta, Leptospira Gram-negative.
(some species)
Magnetospirillum Magnetic crystals allow them to move in water and sediments. Proteobacteria
Gram-negative.
Spirillum Spiral-shaped, microaerophilic bacteria. Gram-negative. Proteobacteria
Sulfur-oxidizing, nitrate-reducing Use novel mechanisms to compensate for the fact that their energy source Proteobacteria
marine bacteria—Thioploca, (reduced sulfur compounds) and terminal electron acceptor (nitrate) do not
Thiomargarita coexist.

Animals as Habitats—See table 11.3

Clostridium species, which are obligate anaerobes, were discussed

earlier. Bacillus species include both obligate aerobes and faculta-
tive anaerobes, and some are medically important. B. anthracis
causes the disease anthrax, which can be acquired from contacting
its endospores in soil or in animal hides or wool. Unfortunately,
the spores have also been used as an agent of domestic bioterror-
ism. endospores, p. 67 anthrax, p. 453

The Genus Azotobacter

Azotobacter species are Gram-negative pleomorphic, rod-shaped
(a) 1 μm (b) 1 μm
bacteria that live in soil. They can form a type of resting cell called
a cyst (figure 11.15). These have negligible metabolic activity and FIGURE 11.15 Azotobacter (a) Vegetative cells. (b) Cyst.
can withstand drying and ultraviolet radiation but are not highly ? What types of adverse environmental conditions can Azotobacter
resistant to heat. cysts withstand?
268 Chapter 11 The Diversity of Prokaryotic Organisms

Azotobacter species are also notable for their ability to fix

nitrogen in aerobic conditions; recall that the enzyme nitrogenase
is inactivated by O2. Apparently, the exceedingly high respiratory
rate of Azotobacter species consumes O2 so rapidly that a low O2
environment is maintained inside the cell. In addition, a protein in
the cell binds nitrogenase, thereby protecting it from O2 damage.

Myxobacteria
The myxobacteria are a group of aerobic Gram-negative rods that
have a unique developmental cycle as well as a resting stage. When
conditions are favorable, cells secrete a slime layer that other cells
then follow, creating a swarm of cells. But then, when nutrients are
exhausted, the behavior of the group changes. The cells begin to con-
gregate, and then pile up to form a complex structure called a fruiting
body, which is often brightly colored (figure  11.16). In some spe-
cies, the fruiting body is quite elaborate, consisting of a mass of cells
elevated and supported by a stalk made of a hardened slime. The cells
within the fruiting body differentiate to become spherical, dormant 5 μm
forms called microcysts. These are considerably more resistant to heat, FIGURE 11.17 Streptomyces A photomicrograph showing the
drying, and radiation than are the vegetative cells of myxobacteria but spherical conidia at the ends of the filamentous hyphae.
are much less resistant than bacterial endospores. ? What is a mass of hyphae called?
Myxobacteria are important in nature as degraders of complex
organic substances; they can digest bacteria and certain algae and
fungi. Scientifically, these bacteria serve as an important model Streptomyces species produce a variety of extracellular
for studying developmental biology. Included in the myxobacteria enzymes that allow them to degrade various organic compounds.
are the genera Chondromyces, Myxococcus, and Stigmatella. They are also responsible for the characteristic “earthy” odor of
soil; like the cyanobacteria, they produce geosmin. One species of
The Genus Streptomyces Streptomyces, S. somaliensis, can cause an infection of subcutane-
The genus Streptomyces encompasses more than 500 species of ous tissue called an actinomycetoma.
aerobic Gram-positive bacteria that resemble fungi in their pattern Streptomyces species naturally produce a wide array of medi-
of growth. Like the fungi, they form a mycelium (a visible mass of cally useful antibiotics, including streptomycin, tetracycline, and
branching filaments). The filaments are called hyphae. Chains of erythromycin. The role these compounds play in the life cycle of
characteristic spores called conidia develop at the tips of hyphae Streptomyces is not entirely understood, but at the low levels pro-
(figure 11.17). These dormant spores are resistant to drying and duced in soils, they appear to be involved in cell signaling.
are easily spread in air currents. Note that even though this pattern
of growth resembles fungi, which are eukaryotes, Streptomyces
species are much smaller and are prokaryotes. Bacteria That Associate with Plants
Members of two related genera use very different means to obtain
nutrients from plants. Agrobacterium species are plant pathogens
that cause tumorlike growths, whereas Rhizobium species form a
mutually beneficial relationship with certain types of plants.

The Genus Agrobacterium

Agrobacterium species are Gram-negative rod-shaped
bacteria that have an unusual mechanism of gaining
a competitive advantage in soil. They cause plant
tumors, the outcome of their ability to genetically
alter plants for their own benefit (figure  11.18).
They do this by attaching to wounded plant tis-
sue, and then transferring a portion of a plasmid
(a) (b) to a plant cell; in A. tumefaciens the plasmid is
called the Ti plasmid (for “tumor-inducing”).
FIGURE 11.16 Fruiting Bodies of
Myxobacteria These are the elaborate fruiting The transferred DNA encodes the ability to syn-
bodies of a species of Chondromyces: (a) photograph; thesize plant growth hormones, causing uncon-
(b) scanning electron micrograph. trolled growth of the plant tissue and resulting in
? How do fruiting bodies help myxobacteria survive a tumor. The transferred DNA also encodes enzymes
adverse conditions? that direct the synthesis of an opine, an unusual amino
 Part II The Microbial World 269

(a)

FIGURE 11.18 Plant Tumor Caused by Agrobacterium

tumefaciens
? What is the role of the Ti plasmid in plant tumor formation?

acid derivative; Agrobacterium can then use this compound as a

nutrient source (see Perspective 8.2).

MicroByte
Scientists have modified the Ti plasmid, turning it into a commercially
valuable tool used to genetically engineer plant cells.
(b) 5 μm

Rhizobia
FIGURE 11.19 Symbiotic Relationship Between Rhizobia
Rhizobia are a group of Gram-negative rod-shaped bacteria that and Certain Plants (a) Root nodules. (b) Scanning electron micro-
often fix nitrogen and form intimate relationships with legumes graph of bacterial cells within a nodule.
(plants that bear seeds in pods). This group of bacteria includes ? How do rhizobia benefit plants?
members of the genera Rhizobium, Sinorhizobium, Bradyrhizobium,
Mesorhizobium, and Azorhizobium. The bacteria live within cells in
nodules formed on the roots of the plants (figure 11.19). The plants
synthesize the protein leghemoglobin, which binds and controls
the levels of O2 (see figure 29.13). Within the resulting microaero-
bic environment of nodules, the bacteria are able to fix nitrogen.
11.7 ■ Thriving in Aquatic
Rhizobia residing within plant cells are examples of endosymbi- Environments
onts, organisms that provide a benefit to the cells in which they
reside. symbiotic nitrogen fixers, p. 730 Learning Outcome
9. Describe the examples of the four mechanisms aquatic bacteria use
MicroAssessment 11.6 to maximize nutrient acquisition and retention.

Bacillus and Clostridium species make endospores, the most resistant

Most aquatic environments lack a steady supply of nutrients. To
type of dormant cell known. Azotobacter species, myxobacteria,
and Streptomyces species all produce dormant cells that tolerate thrive in these habitats, bacteria have evolved various mechanisms
some adverse conditions but are less resistant than endospores. to maximize nutrient acquisition and retention.
Agrobacterium species and rhizobia obtain nutrients from plants, but
the former are plant pathogens and the latter benefit the plant.
16. Why are myxobacteria important in nature?
Sheathed Bacteria
17. How does Agrobacterium benefit from inducing a plant tumor? Sheathed bacteria form chains of cells encased within a tube, or
18. If you wanted to determine the number of endospores in a sample
sheath (figure  11.20). This plays a protective role, helping the
of soil, what could you do before plating it? + bacteria attach to solid objects located in favorable habitats while
sheltering them from attack by predators. Masses of filamentous
270 Chapter 11 The Diversity of Prokaryotic Organisms

Prosthecate Bacteria
The prosthecate bacteria are a diverse group of Gram-negative
bacteria that have projections called prosthecae, which are exten-
sions of the cytoplasm and cell wall. These extensions provide
increased surface area to facilitate absorption of nutrients. Some
prosthecae allow the organisms to attach to solid surfaces.

The Genus Caulobacter

Because of their remarkable life cycle, Caulobacter species serve
as a model for research on cellular differentiation. Entirely differ-
ent events occur in an orderly fashion at opposite ends of the cell.
Caulobacter cells have a single polar prostheca, commonly
called a stalk (figure  11.21). At the tip of the stalk is an adhe-
sive holdfast, which provides a mechanism for attachment. To
multiply, the cell elongates and divides by binary fission, pro-
Sheath Bacterial cells 10 μm ducing a motile swarmer cell at the end opposite the stalk. This
FIGURE 11.20 Sheathed Bacteria Phase-contrast photomicro- swarmer cell has a flagellum, located at the pole opposite the site
graph of a Sphaerotilus species.
of division. The swarmer cell detaches and moves to a new loca-
? What is the role of the sheath? tion, where it adheres via a holdfast near the base of its flagellum.
It then loses its flagellum, replacing it with a stalk. Only then
can the daughter cell replicate its DNA and repeat the process. In
sheaths can often be seen streaming from rocks in flowing water favorable conditions, a single cell divides and produces daughter
polluted by nutrient-rich wastes. They often interfere with sew- cells many times. With each division, a ring remains at the site of
age treatment and other industrial processes by clogging pipes. division, allowing a researcher to count the number of progeny.
Sheathed bacteria include species of Sphaerotilus and Leptothrix,
which are Gram-negative rods. The Genus Hyphomicrobium
Sheathed bacteria spread by forming motile cells called Hyphomicrobium species are in many ways similar to
swarmer cells that exit through the unattached end of the sheath. Caulobacter species, except they have a distinct method of
These then move to a new solid surface, where they attach. If reproduction. The single polar prostheca of the parent cell
enough nutrients are present, they can multiply and form a new enlarges at the tip to form a bud (figure 11.22). This continues
sheath, which gets longer as the chain of cells grows. enlarging and develops a flagellum, eventually giving rise to

Flagellum Swarmer cell

Flagellum
shedding

Separating Holdfast
daughter cell

Stalked cell

Stalk
Stalk formation

Dividing
Holdfast bacterium

(a) 1 μm (b)

FIGURE 11.21 Caulobacter (a) Photomicrograph. (b) Life cycle.

? What characteristic of Caulobacter species makes them important research models?
 Part II The Microbial World 271

Swarmer cell
Young with a flagellum
bud
New nucleoid
moving into
hypha
Hypha
forming

Nucleoid

(a) 1 μm

Hypha lengthens more and produces another bud.

(b)
FIGURE 11.22 Hyphomicrobium (a) Photomicrograph. Note the bud forming at the tip of the polar prostheca. (b) Life cycle.
? What will happen to the swarmer cell?

a motile daughter cell. The daughter cell (swarmer cell) then The Genus Bdellovibrio
detaches and moves to a new location, eventually losing its
Bdellovibrio species (bdello, from the Greek word for “leech”) are
flagellum and forming a polar prostheca at the opposite end to
highly motile Gram-negative curved rods that prey on E. coli and
repeat the cycle. As with Caulobacter species, a single cell can
other Gram-negative bacteria (figure 11.23). When a Bdellovibrio
repeatedly produce daughter cells.
cell attacks, it strikes its prey with such force that it propels the
prey a short distance. The parasite then attaches to its host and
Bacteria That Derive Nutrients rotates with a spinning motion. At the same time, it makes diges-
from Other Organisms tive enzymes that break down lipids and peptidoglycan, eventu-
Some bacteria obtain nutrients directly from other organisms. ally forming a hole in the cell wall of the prey. This allows the
Examples include Bdellovibrio species, bioluminescent bacteria, parasitic bacterium to penetrate the peptidoglycan, lodging in the
Epulopiscium species, and Legionella species. periplasm. There, over a period of several hours, Bdellovibrio

Bacterial prey
Bdellovibrios

Bacterial prey

10 min

Cell wall
20 min Cytoplasmic
membrane

10 sec

Bdellovibrio
150–210 min

20 min
(a) 1 μm (b)

FIGURE 11.23 Bdellovibrio (a) Color-enhanced transmission micrograph of a Bdellovibrio cell attacking its prey. (b) Life cycle of
Bdellovibrio. Note that the diagram exaggerates the size of the space in which Bdellovibrio multiplies.
? How does a Bdellovibrio cell penetrate the prey?
272 Chapter 11 The Diversity of Prokaryotic Organisms

degrades and utilizes the prey’s cellular contents. It derives energy cell has thousands of copies of the genome dispersed throughout
by aerobically oxidizing amino acids and acetate. The parasite the cell. The multiple genome copies might help the organism
increases in length, ultimately dividing to form several motile overcome the problem of ensuring that necessary proteins are
daughter cells. When the host cell lyses, the Bdellovibrio progeny synthesized even in the far reaches of the large cell.
are released to find new hosts, repeating the cycle. periplasm, p. 61 Epulopiscium species have an unusual life cycle. Rather than
undergoing typical binary fission, they enlarge considerably,
Bioluminescent Bacteria finally lysing to release up to seven daughter cells. They have not
Some species of Photobacterium and Vibrio can emit light yet been grown in culture.
(figure  11.24). This phenomenon, bioluminescence, plays an
important role in the symbiotic relationship between some of these The Genus Legionella
bacteria and specific types of fish and squid. For example, cer- Legionella species are commonly found in aquatic environments,
tain types of squid have a specialized organ colonized by Vibrio where they often reside within protozoa. They have even been
fischeri within their ink sac. The light produced in the organ is isolated from water in air conditioners and produce misters. They
thought to serve as a type of camouflage, obscuring the squid’s are Gram-negative obligate aerobes that use amino acids, but
contrast against the light from above and any shadow it might not carbohydrates, as a source of carbon and energy. Legionella
otherwise cast. The squid provides nutrients to the symbiotic bac- pneumophila can cause respiratory disease when inhaled in aero-
teria, facilitating their growth. Another example is the flashlight solized droplets. Legionnaires’ disease, p. 506
fish, which has a light organ harboring bioluminescent bacteria
in a specialized pouch below its eye. By opening and closing a
lid that covers the pouch, the fish can control the amount of light Bacteria That Move
released, a tactic believed to confuse predators and prey. by Unusual Mechanisms
Luminescence is catalyzed by the enzyme luciferase. Studies Some bacteria have unique mechanisms of motility that allow
revealed that the genes encoding it are expressed only when the them to easily move to desirable locations. These organisms
density of the bacterial population reaches a critical point. This include the spirochetes and the magnetotactic bacteria.
phenomenon of quorum sensing is now recognized as an impor-
tant mechanism by which a variety of different bacteria regulate Spirochetes
the expression of certain genes. quorum sensing, p. 177 The spirochetes (Greek spira for “coil” and chaete for “hair”) are a
Photobacterium and Vibrio species are Gram-negative rods group of Gram-negative bacteria with a unique motility mechanism
(Vibrio species are curved rods) with polar flagella. They are that allows them to move through thick, viscous environments such as
facultative anaerobes and typically inhabit aqueous environments; mud. Distinguishing characteristics include their spiral shape, flexible
species that require sodium are found in marine environments. cell wall, and motility by means of endoflagella (also called an axial
Not all are luminescent, and some Vibrio species cause human filament). Unlike typical flagella, endoflagella are contained within
disease. Pathogens include V. cholerae, which causes cholera, and the periplasm. Either a single flagellum or a tuft originates at each
V. parahaemolyticus, which also causes diarrheal disease; neither end of the cell, and the flagella extend toward each other, overlapping
of these is bioluminescent. Vibrio cholerae, p. 586 in the mid-region of the cell. Rotation of the endoflagella within the
confines of the periplasm causes the cell to move like a corkscrew,
The Genus Epulopiscium sometimes deviating into flexing motions. Many spirochetes are very
Epulopiscium species are Gram-positive cigar-shaped bacteria slender and can be seen only by using special methods such as dark-
that reside in the intestinal tract of surgeon fish. They are consid- field microscopy (figure 11.25). Many are also difficult or impossible
erably larger than most prokaryotes (600 μm × 80 μm), and each to grow in culture. periplasm, p. 61
Spirochetes include free-living species that inhabit aquatic
environments, as well as ones that reside on or in animals.
Spirochaeta species are anaerobes or facultative anaerobes that
thrive in muds and anaerobic waters. Leptospira species are aero-
bic; some are free-living in aquatic environments, whereas others
grow within animals. L. interrogans causes the disease leptospi-
rosis, which can be transmitted in the urine of infected animals.
Spirochetes adapted to reside in body fluids of humans and other
animals will be discussed later. leptospirosis, p. 615

Magnetotactic Bacteria
(a) (b)
Magnetotactic bacteria such as Magnetospirillum (Aquaspirillum)
FIGURE 11.24 Luminescent Bacteria (a) Plate culture of biolumi- magnetotacticum contain a string of magnetic crystals that align
nescent bacteria. (b) Photograph of a flashlight fish; under the eye is a cells with the earth’s magnetism (see figure  3.39). This allows
light organ colonized with bioluminescent bacteria. them to move up or down in the water or sediments. It is thought
? What role does quorum sensing play in bioluminescence? that this unique type of movement allows them to locate the
 Part II The Microbial World 273

Spirochetes due to globules of sulfur in their cytoplasm (see Perspective 1.1).

Each cell contains a large nitrate storage vacuole that takes up about
98% of the cell volume. These organisms, which can reach a diam-
eter of 0.75 mm, are not motile and instead rely on storms or other
disturbances to bring them into contact with nitrate-rich waters.

MicroByte
Thiomargarita namibiensis can store a 3-month supply of both its
energy source (sulfur) and its terminal electron acceptor (nitrate).

MicroAssessment 11.7
Sheathed bacteria attach to solid objects in favorable locations.
Prosthecate bacteria produce extensions that maximize the absorptive
surface area. Bdellovibrio species, bioluminescent bacteria, and
Legionella species use nutrients from other organisms. Spirochetes
5 μm and magnetotactic bacteria move by unusual mechanisms. Some
FIGURE 11.25 Spirochetes Dark-field photomicrograph of organisms form storage granules.
spirochetes. 19. How do squid benefit from having a light organ colonized by
? Why are members of this genus difficult to see with bright-field luminescent bacteria?
microscopy? 20. What is the habitat of Legionella species?
21. The genomes of free-living spirochetes are larger than those
microaerophilic habitats they require. Magnetospirillum species of ones that live within an animal host. Why would this be so? +
are Gram-negative, spiral-shaped organisms. magnetotaxis, p. 64

Bacteria That Form Storage Granules 11.8 ■ Animals as Habitats

A number of aquatic bacteria form granules that store nutrients. Learning Outcome
Recall that anoxygenic phototrophs often store sulfur granules,
10. Compare and contrast the examples of bacteria that use animals
which can later be used as a source of electrons for reducing as habitats.
power. Some bacteria store phosphate, and others store com-
pounds that can be used to generate ATP. The bodies of animals, including humans, provide a wide variety
of ecological habitats for microbes—from dry, O2-rich surfaces to
The Genus Spirillum moist, anaerobic recesses. Table  11.3 lists the medically impor-
Spirillum species are Gram-negative spiral-shaped, microaero- tant bacteria covered in this and other sections of the chapter.
philic bacteria. Spirillum volutans forms volutin granules, which
are storage forms of phosphate. These are sometimes called meta-
chromatic granules to reflect their characteristic staining with the Bacteria That Inhabit the Skin
dye methylene blue. The cells of S. volutans are typically large, The skin is typically dry and salty, providing an environment
over 20 μm in length. In wet mounts, Spirillum species may be inhospitable to many microorganisms. Members of the genus
seen moving to a narrow zone near the edge of the coverslip, Staphylococcus, however, thrive under these conditions. The
where O2 is available in the optimum amount. volutin, p. 67 propionic acid bacteria, which were discussed earlier, inhabit
anaerobic microenvironments of the skin. anatomy, physiology, and
Sulfur-Oxidizing, Nitrate-Reducing ecology of the skin, p. 522
Marine Bacteria
Some marine bacteria store both sulfur (their energy source) and The Genus Staphylococcus
nitrate (their terminal electron acceptor). This provides an advan- Staphylococcus species are Gram-positive cocci that are faculta-
tage to the bacteria, because reduced sulfur compounds are often tive anaerobes. Most, such as S. epidermidis, reside harmlessly as
abundant in anaerobic marine sediments, but these environments part of the normal microbiota of the skin. Like other bacteria that
lack suitable terminal electron acceptors. In contrast, waters above aerobically respire, Staphylococcus species are catalase-positive.
those sediments lack reduced sulfur compounds but provide a This distinguishes them from Streptococcus, Enterococcus, and
source of nitrate. In other words, the energy source and terminal Lactococcus species, which are also Gram-positive cocci but
electron acceptor do not coexist. To cope with this, Thioploca lack the enzyme catalase. Several species of Staphylococcus are
species form long sheaths within which cells shuttle between the notable for their medical significance. Staphylococcus aureus
sulfur-rich sediments and nitrate-rich waters, storing reserves of causes a variety of diseases, including skin and wound infections,
sulfur and nitrate. The cells of the huge bacterium Thiomargarita as well as food poisoning. Staphylococcus saprophyticus causes
namibiensis (“sulfur pearl of Namibia”) are a pearly white color urinary tract infections.
274 Chapter 11 The Diversity of Prokaryotic Organisms

TABLE 11.3 Medically Important Bacteria

Organism Medical Significance Phylum

Gram-Negative Rods
Bacteroides species Obligate anaerobes that commonly inhabit the mouth, intestinal tract, and Bacteroidetes
genital tract. Cause abscesses and bloodstream infections.
Enterobacteriaceae Proteobacteria
Enterobacter species Normal microbiota of the intestinal tract.
Escherichia coli Normal microbiota of the intestinal tract. Some strains cause urinary tract
infections; some strains cause specific types of intestinal disease; some cause
meningitis in newborns.
Klebsiella pneumoniae Normal microbiota of the intestinal tract. Causes pneumonia.
Proteus species Normal microbiota of the intestinal tract. Cause urinary tract infections.
Salmonella enterica serotype Causes gastroenteritis. Grows in the intestinal tract of infected animals;
Enteritidis acquired by consuming contaminated food.
Salmonella enterica serotype Causes typhoid fever. Grows in the intestinal tract of infected humans;
Typhi transmitted in feces.
Shigella species Cause dysentery. Grow in the intestinal tract of infected humans; transmitted
in feces.
Yersinia pestis Causes bubonic plague, which is transmitted by fleas, and pneumonic
plague, which is transmitted in respiratory droplets of infected individuals.
Haemophilus influenzae Causes ear infections, respiratory infections, and meningitis in children. Proteobacteria
Haemophilus ducreyi Causes chancroid, a sexually transmitted disease. Proteobacteria
Legionella pneumophila Causes Legionnaires’ disease, a lung infection. Grows within protozoa; Proteobacteria
acquired by inhaling contaminated water droplets.
Pseudomonas aeruginosa Causes burn, urinary tract, and bloodstream infections. Common in the Proteobacteria
environment. Grows in nutrient-poor aqueous solutions. Resistant to many
disinfectants and antimicrobial medications.
Gram-Negative Rods—Obligate Intracellular Parasites
Chlamydophila (Chlamydia) Causes atypical pneumonia, or “walking pneumonia.” Acquired from an Chlamydiae
pneumoniae infected person.
Chlamydophila (Chlamydia) Causes psittacosis, a form of pneumonia. Transmitted by birds. Chlamydiae
psittaci
Chlamydia trachomatis Causes a sexually transmitted disease that mimics the symptoms of Chlamydiae
gonorrhea. Also causes trachoma, a serious eye infection, and conjunctivitis
in newborns.
Coxiella burnetii Causes Q fever. Acquired by inhaling organisms shed by infected animals. Proteobacteria
Ehrlichia chaffeensis Causes human ehrlichiosis. Transmitted by ticks. Proteobacteria
Orientia tsutsugamushi Causes scrub typhus. Transmitted by mites. Proteobacteria
Rickettsia prowazekii Causes epidemic typhus. Transmitted by lice. Proteobacteria
Rickettsia rickettsii Causes Rocky Mountain spotted fever. Transmitted by ticks. Proteobacteria
Wolbachia pipientis Resides within the filarial worms that cause river blindness and elephantiasis. Proteobacteria
Gram-Negative Curved Rods
Campylobacter jejuni Causes gastroenteritis. Grows in the intestinal tract of infected animals; Proteobacteria
acquired by consuming contaminated food.
Helicobacter pylori Causes stomach and duodenal ulcers. Neutralizes stomach acid by producing Proteobacteria
urease.
Vibrio cholerae Causes cholera, a severe diarrheal disease. Grows in the intestinal tract of Proteobacteria
infected humans; acquired by drinking contaminated water.
Vibrio parahaemolyticus Causes gastroenteritis. Acquired by consuming contaminated seafood. Proteobacteria
Gram-Negative Cocci
Neisseria meningitidis Causes meningitis. Proteobacteria
Neisseria gonorrhoeae Causes gonorrhea, a sexually transmitted infection. Proteobacteria

(continued)
 Part II The Microbial World 275

TABLE 11.3 Medically Important Bacteria (Continued)

Organism Medical Significance Phylum

Gram-Positive Rods
Bacillus anthracis Causes anthrax. Acquired by inhaling endospores in soil, animal hides, and Firmicutes
wool. Bioterrorism agent.
Bifidobacterium species Predominant member of the intestinal tract in breast-fed infants. Thought to Actinobacteria
play a protective role in the intestinal tract by excluding pathogens.
Clostridium botulinum Causes botulism. Disease results from ingesting toxin-contaminated foods, Firmicutes
typically canned foods that have been improperly processed.
Clostridium perfringens Causes gas gangrene. Acquired when soil-borne endospores contaminate a Firmicutes
wound.
Clostridium tetani Causes tetanus. Acquired when soil-borne endospores are inoculated into Firmicutes
deep tissue.
Corynebacterium diphtheriae Toxin-producing strains cause diphtheria, a frequently fatal throat infection. Actinobacteria
Gram-Positive Cocci
Enterococcus species Normal microbiota of the intestinal tract. Cause urinary tract infections. Firmicutes
Micrococcus species Found on skin as well as in a variety of other environments; often Actinobacteria
contaminate bacteriological media.
Staphylococcus aureus Leading cause of wound infections. Causes boils, carbuncles, food poisoning, Firmicutes
and toxic shock syndrome.
Staphylococcus epidermidis Normal microbiota of the skin. Firmicutes
Staphylococcus saprophyticus Causes urinary tract infections. Firmicutes
Streptococcus pneumoniae Causes pneumonia and meningitis. Firmicutes
Streptococcus pyogenes Causes pharyngitis (strep throat), rheumatic fever, wound infections, Firmicutes
glomerulonephritis, and streptococcal toxic shock.
Acid-Fast Rods
Mycobacterium tuberculosis Causes tuberculosis. Actinobacteria
Mycobacterium leprae Causes Hansen’s disease (leprosy); peripheral nerve invasion is characteristic. Actinobacteria
Spirochetes
Treponema pallidum Causes syphilis, a sexually transmitted disease. The organism has never been Spirochaetes
grown in culture.
Borrelia burgdorferi Causes Lyme disease, a tick-borne disease. Spirochaetes
Borrelia recurrentis and B. hermsii Causes relapsing fever. Transmitted by arthropods. Spirochaetes
Leptospira interrogans Causes leptospirosis, a waterborne disease. Excreted in urine of infected Spirochaetes
animals.
Cell Wall-less
Mycoplasma pneumoniae Causes atypical pneumonia (“walking pneumonia”). Not susceptible to Tenericutes
penicillin because it lacks a cell wall.

Bacteria That Inhabit genital tract of humans and other animals. Bacteroides fragilis
Mucous Membranes and related species make up about a third of the bacteria in human
feces and are often responsible for abscesses and bloodstream
Mucous membranes of the respiratory, genitourinary, and intesti- infections that follow appendicitis and abdominal surgery. Many
nal tracts provide a habitat for numerous kinds of bacteria, many are killed by brief exposure to O2, so they are difficult to study.
of which have already been discussed. For example, Streptococcus
and Corynebacterium species reside in the respiratory tract, The Genus Bifidobacterium
Lactobacillus species inhabit the vagina, and Clostridium species Bifidobacterium species are Gram-positive, irregular, rod-shaped
and members of the family Enterobacteriaceae thrive in the intes- anaerobes that reside primarily in the intestinal tract of humans
tinal tract. Some of the other genera are discussed next. and other animals. They are the predominant members of the
intestinal microbiota of breast-fed infants and are thought to pro-
The Genus Bacteroides vide a protective function by excluding disease-causing bacteria.
Bacteroides species are small, strictly anaerobic, Gram-negative Formula-fed infants are also colonized with members of this
rods and coccobacilli. They inhabit the mouth, intestinal tract, and genus, but generally the concentrations are lower.
276 Chapter 11 The Diversity of Prokaryotic Organisms

The Genera Campylobacter

and Helicobacter
Members of the genera Campylobacter and Helicobacter
are curved Gram-negative rods. As microaerophiles, they
require specific atmospheric conditions to grow in culture.
Campylobacter jejuni causes diarrheal disease in humans.
It typically lives in the intestinal tract of domestic animals,
particularly poultry. Helicobacter pylori inhabits the stomach,
where it can cause stomach and duodenal ulcers; it has also
been linked to stomach cancer. An important factor in its abil-
ity to survive in the stomach is its production of the enzyme
urease. This breaks down urea to produce ammonia, which
neutralizes the acid in the cell’s immediate surroundings.
stomach ulcers, p. 580

The Genus Haemophilus

Haemophilus species are Gram-negative coccobacilli that, as
their name reflects, are “blood loving.” They require hematin
and/or NAD, which are found in blood. Many species are com- FIGURE 11.26 Mycoplasma pneumoniae Colonies Note the
mon microbiota of the respiratory tract. H. influenzae causes dense central portion of the colony, giving it the typical “fried egg”
ear infections, respiratory infections, and meningitis, primar- appearance.
ily in children. Haemophilus ducreyi causes the sexually ? Why are members of this genus not affected by penicillin?
transmitted disease chancroid. ear infections, p. 492 meningitis,
p. 643 chancroid, p. 629

The Genera Treponema and Borrelia

The Genus Neisseria
Members of the genera Treponema and Borrelia are spirochetes
Neisseria species are Gram-negative bacteria, typically kidney- that typically inhabit body fluids and mucous membranes of
bean-shaped cocci in pairs. They are common microbiota of humans and other animals. Recall that spirochetes are character-
animals including humans, growing on mucous membranes. ized by their corkscrew shape and endoflagella. Although they
Neisseria species are typically aerobes, but some can grow anaer- have a Gram-negative cell wall, they are often too thin to be
obically if a suitable terminal electron acceptor such as nitrite viewed by conventional microscopy.
is present. Those noted for their medical significance include Treponema species are obligate anaerobes or microaero-
N. gonorrhoeae, which causes the sexually transmitted disease philes that often inhabit the mouth and genital tract. Study
gonorrhea, and N. meningitidis, which causes meningitis; both of the species that causes syphilis, T. pallidum, is difficult
are  nutritionally fastidious. fastidious, p.  93 gonorrhea, p.  622
because it has never been grown in culture. Its genome has
meningitis, p. 643
been sequenced, however, providing evidence that it is a
microaerophile with a metabolism highly dependent on its
The Genus Mycoplasma host. It lacks critical enzymes of the TCA cycle and a variety
Members of the genus Mycoplasma lack a cell wall, making them of other metabolic pathways.
flexible and able to pass through the pores of filters that retain Three Borrelia species are pathogens, transmitted by
other bacteria. Most have sterols in their membrane, providing arthropods such as ticks and lice. B. recurrentis and B. hermsii
added strength and rigidity, thereby protecting the cells from both cause relapsing fever; B. burgdorferi causes Lyme dis-
osmotic lysis. They are among the smallest forms of life, and their ease. A striking feature of Borrelia species is their genome—
genomes are thought to be the minimum size for encoding the a linear chromosome and many linear and circular plasmids.
essential functions for a free-living organism.
Medically, the most significant member of this group is
M. pneumoniae, which, as its name implies, causes a form of pneu- Obligate Intracellular Parasites
monia. This type of pneumonia, often called “walking pneumonia,” Obligate intracellular parasites cannot reproduce outside
cannot be treated with penicillin or other antibiotics that interfere a host cell. By living within host cells, the parasites are sup-
with peptidoglycan synthesis, because these organisms lack a cell plied with a readily available source of compounds they would
wall. Colonies of Mycoplasma species growing on solid media otherwise need to synthesize for themselves. As a result,
produce a characteristic “fried egg” appearance (figure 11.26). most intracellular parasites have lost the ability to make sub-
stances needed for extracellular growth. Bacterial examples
MicroByte
include members of the genera Rickettsia, Orientia, Ehrlichia,
The genome of Mycoplasma genitalium is only 5.8 ×105 base pairs,
approximately one-eighth the size of the E. coli genome. Coxiella, Chlamydia, and Wolbachia, which are all tiny Gram-
negative rods or coccobacilli.
 Part II The Microbial World 277

The Genera Rickettsia, Orientia, and Ehrlichia

Species of Rickettsia, Orientia, and Ehrlichia are responsible for
several serious human diseases spread by blood-sucking arthro-
pods such as ticks and lice. Rickettsia rickettsii causes Rocky
Mountain spotted fever, R. prowazekii causes epidemic typhus,
O. tsutsugamushi causes scrub typhus, and E. chaffeensis causes
human ehrlichiosis.

The Genus Coxiella

The only characterized species of Coxiella, C. burnetii, is an
obligate intracellular bacterium that survives well outside the
host cell. During its intracellular growth, C. burnetii forms spore-
like structures called small-cell variants (SCVs) that later allow
it to survive in the environment. The structures, however, lack
the extreme resistance to heat and disinfectants characteristic of
endospores (figure  11.27). Coxiella burnetii causes Q fever of 0.2 μm
humans, a disease most often acquired by inhaling bacteria shed FIGURE 11.28 Chlamydia Growing in Tissue Cell Culture The
from infected animals. This is particularly a problem with preg- numbers indicate the development from the dividing reticulate body
nant animals because high numbers of the bacteria can be found to an infectious elementary body.
in the placenta of infected animals. ? How is the cell wall of Chlamydia highly unusual?

The Genera Chlamydia and Chlamydophila

Chlamydia and Chlamydophila species are quite different from
The Genus Wolbachia
the other obligate intracellular parasites. They are transmitted
directly from person to person, and have a unique growth cycle The only known species of Wolbachia, W. pipientis, infects arthro-
(figure  11.28). Inside the host cell, they initially exist as non- pods (including insects, spiders, and mites) and parasitic worms.
infectious reticulate bodies, which reproduce by binary fission. It is primarily transmitted maternally, via the eggs of infected
Later in the infection, the bacteria differentiate into smaller, females to their offspring. In arthropods, the bacterium uses
dense-appearing infectious elementary bodies, which are released unique strategies to increase the overall population of infected
when the host cell ruptures. The cell wall of Chlamydia and females, including killing male embryos, allowing infected
Chlamydophila species lacks peptidoglycan, but it has the other females to reproduce asexually, and causing infected males to gain
features of a Gram-negative type of cell wall. Chlamydia trachomatis female traits. In addition, the parasite destroys embryos resulting
causes eye infections and a sexually transmitted infection that mimics from the mating of an infected male with either an uninfected
gonorrhea; Chlamydophila pneumoniae causes atypical pneumonia; female or a female infected with a different strain. Wolbachia does
and Chlamydophila psittaci causes psittacosis, a form of pneumonia. not infect mammals, but its medical importance was recognized
Chlamydia trachomatis, p. 625
with the discovery that it resides within filarial worms that cause
the diseases river blindness and elephantiasis. The chronic and
debilitating inflammation associated with these diseases appears
to result from the immune response directed against the bacterial
cells in the invading worms. This discovery paves the way for new
treatments, because eliminating the bacteria not only lessens the
symptoms but also kills the filarial worms.

MicroAssessment 11.8
Staphylococcus species thrive in the dry, salty conditions of the skin.
Bacteroides and Bifidobacterium species reside in the gastrointestinal
tract; Campylobacter and Helicobacter species can cause disease
when they reside there. Neisseria species, mycoplasmas, and
spirochetes inhabit other mucous membranes. Obligate intracellular
parasites—including Rickettsia, Orientia, Ehrlichia, Coxiella,
Chlamydia, Chlamydophila, and Wolbachia species—are unable to
reproduce outside of a host cell.
22. How does Helicobacter pylori withstand stomach acidity?
0.1 μm
23. What characteristic of Mycoplasma species separates them
FIGURE 11.27 Coxiella Color-enhanced transmission electro- from other bacteria?
micrograph of C. burnetii. The oval copper-colored object is
the sporelike structure. 24. Why would breast feeding affect the composition of a
baby’s intestinal microbiota? +
? What disease does C. burnetii cause?
278 Chapter 11 The Diversity of Prokaryotic Organisms

11.9 ■ Archaea That Thrive

in Extreme Conditions
Learning Outcome
11. Compare and contrast the characteristics and habitats of the
extreme halophiles and extreme thermophiles.

Characterized members of the Archaea typically thrive in extreme

environments (table 11.4). These include conditions of high heat,
acidity, alkalinity, and salinity. An exception is the methanogens,
discussed earlier in the chapter; they inhabit anaerobic niches
shared with members of the Bacteria. In addition to the character-
ized archaea, many others have been detected in a variety of non-
extreme environments using molecular techniques.
FIGURE 11.29 Solar Evaporation Pond
? What causes the pink color in the pond?
Extreme Halophiles
The extreme halophiles are found in high numbers in salty envi-
ronments such as salt lakes, soda lakes, and brines used for curing Methane-Generating Hyperthermophiles
fish. Most grow well in saturated salt solutions (32% NaCl), and In contrast to the mesophilic methanogens discussed earlier,
they require a minimum of about 9% NaCl. Because they produce some methanogens are extreme thermophiles. For example,
pigments, their growth can be seen as red patches on salted fish Methanothermus species, which can grow in temperatures as high
and pink blooms in concentrated salt water ponds (figure 11.29). as 97°C, grow optimally at approximately 84°C. Like all methano-
Extreme halophiles are aerobic or facultatively anaerobic gens, they oxidize H2, using CO2 as a terminal electron acceptor
chemoheterotrophs, but some also can obtain additional energy to yield methane.
from light. These organisms have the light-sensitive pigment
bacteriorhodopsin, which absorbs energy from sunlight and uses it Sulfur-Reducing Hyperthermophiles
to expel protons from the cell. This creates a proton gradient that The sulfur-reducing hyperthermophiles are obligate anaerobes
can be used to drive flagella or synthesize ATP. that use sulfur as a terminal electron acceptor, generating H2S.
Extreme halophiles come in a variety of shapes, includ- They harvest energy by oxidizing organic compounds and/or H2.
ing rods, cocci, discs, and triangles. They include genera These archaea can be isolated from hot sulfur-containing environ-
such as Halobacterium, Halorubrum, Natronobacterium, and ments such as sulfur hot springs and hydrothermal vents. They
Natronococcus; members of these latter two genera are extremely include some of the most thermophilic organisms known, a few
alkaliphilic as well as halophilic. even growing above 100°C. One example, Pyrolobus fumarii,
was isolated from a “black smoker” 3,650 m (about 12,000 feet)
deep in the Atlantic Ocean. It grows between 90°C and 113°C.
Extreme Thermophiles Another hydrothermal vent isolate, Pyrodictium occultum, has
The extreme thermophiles (hyperthermophiles) are found near an optimum temperature of about 105°C, and cannot grow below
volcanic vents and fissures that release sulfurous gases and other 82°C. Its disc-shaped cells are connected by hollow tubes, form-
hot vapors. Because these regions are thought to closely mimic ing a weblike network (figure 11.30). The record-holder for the
early earth’s environment, scientists are interested in studying the highest maximum growth temperature, dubbed “strain 121” to
prokaryotes that thrive there. Others are found in hydrothermal reflect that it grows at 121°C, appears to be most closely related
vents in the deep sea and hot springs. hyperthermophiles, p. 90 to Pyrodictium species, but it uses iron as an electron acceptor.

TABLE 11.4 Archaea

Group/Genera Characteristics Phylum

Methanogens—Methanospirillum, Generate methane when they oxidize hydrogen gas as an energy source, Euryarchaeota
Methanosarcina using CO2 as a terminal electron acceptor.
Extreme halophiles—Halobacterium, Found in salt lakes, soda lakes, and brines. Most grow well in saturated salt Euryarchaeota
Halorubrum, Natronobacterium, solutions.
Natronococcus
Extreme thermophiles— Found near hydrothermal vents and in hot springs; some grow at Crenarchaeota,
Methanothermus, Pyrodictium, temperatures above 100°C. Includes examples of methane-generating, Euryarchaeota, and
Pyrolobus, Sulfolobus, Thermophilus, sulfur-reducing, and sulfur-oxidizing archaea, as well as extreme Nanoarchaeota
Picrophilus, Nanoarchaeum acidophiles.
 Part II The Microbial World 279

1 μm
FIGURE 11.31 Typical Habitat of Sulfolobus Sulfur hot spring in
FIGURE 11.30 Pyrodictium The disc-shaped cells are connected
Yellowstone National Park.
by hollow tubes. Scanning electron micrograph.
? How would members of this genus be grouped with respect to
? Why must members of this genus be able to tolerate acidic
conditions?
their temperature preference?
Thermophilic Extreme Acidophiles
Members of two genera, Thermoplasma and Picrophilus, are notable
Nanoarchaea for growing in extremely acidic, hot environments. Thermoplasma
The discovery of an archaeum so unique that it represents an species grow optimally at pH 2; in fact, T. acidophilum lyses at neu-
entirely new phylum, Nanoarcheota (“dwarf archaea”), was made tral pH. It was originally isolated from a coal waste pile. Picrophilus
possible by the earlier discovery of a new genus of sulfur-reducing species tolerate conditions even more acidic, growing optimally at
hyperthermophiles, Ignicoccus (“the fire sphere”). Nanoarchaeum a pH below 1. Two species isolated in Japan inhabited acidic areas
equitans (“rides the fire sphere”) grows as 400-nm spheres in regions that spew sulfurous gases.
attached to the surface of—presumably parasitizing—Ignicoccus
MicroAssessment 11.9
species (see figure 9.20).
Many characterized Archaea inhabit extreme environments. These
Sulfur Oxidizers include conditions of high salinity, heat, acidity, and alkalinity.
Sulfolobus species are found at the surface of acidic sulfur- 25. What is the habitat of Nanoarchaeum equitans?
containing hot springs such as many of those found in Yellowstone 26. At which relative depth in a sulfur hot spring would a sulfur
National Park (figure 11.31). They are obligate aerobes that oxi- oxidizer likely be found?
dize sulfur compounds, using O2 as a terminal electron acceptor 27. What characteristic of the methanogens makes it logical to
to generate sulfuric acid. In addition, they are thermoacidophilic, discuss them with the Bacteria rather than with the Archaea
only growing above 50°C and at a pH between 1 and 6. described in this section? +

FUTURE CHALLENGES 11.1

Astrobiology: The Search for Life on Other Planets
If life as we know it exists on other planets, that brings together scientists from a wide a liquid ocean. Mars is the planet closest to
it will likely be microbial. The task, then, is range of disciplines, including microbiology, earth, and it has the most similar environment.
to figure out how to find and detect such extra- geology, astronomy, biology, and chemistry. Images and data from the recent Mars missions
terrestrial microorganisms. The goal is to determine the origin, evolution, indicate that flowing water once existed there.
Considering that we still know relatively distribution, and destiny of life in the universe. To prepare for researching life on other
little about the microbial life on our own Astrobiologists are also developing light- planets, microbiologists have turned to some
planet, coupled with the extreme difficulty of weight, dependable, and meaningful testing of the most extreme environments here on
obtaining or testing extraterrestrial samples, devices to be used in future space missions. earth. These include glaciers and ice shelves,
this is a daunting challenge with many as yet Astrobiologists believe that within our hot springs, deserts, volcanoes, deep ocean
unanswered questions. What is the most likely solar system, life would most likely be found hydrothermal vents, and subterranean features
source of life on other planets? What is the either on Europa (a moon of Jupiter) or on such as caves. Because select microorganisms
best way to preserve specimens for study on Mars. This is because Europa and Mars appear can survive in these environs, which are analo-
earth? What will be the culture requirements to have, or have had, water—crucial for all gous to conditions expected on other planets,
to grow such organisms? Astrobiology, the known forms of life. Europa has an icy crust, they are good testing grounds for the technol-
study of life in the universe, is a new field beneath which may be liquid water or even ogy to be used on future missions.
280 Chapter 11 The Diversity of Prokaryotic Organisms

Summary
METABOLIC DIVERSITY (table 11.1) PCR. Deinococcus radiodurans can survive radiation exposure several
thousand times that lethal to a human being.
11.1 ■ Anaerobic Chemotrophs
Facultative Anaerobes
Anaerobic Chemolithotrophs
Corynebacterium species are widespread in nature (figure 11.13). Members
The methanogens are a group of archaea that harvest energy by of the family Enterobacteriaceae typically inhabit the intestinal tract of
oxidizing H2, using CO2 as a terminal electron acceptor (figure 11.1). animals, although some reside in rich soil. Coliforms are used as indica-
Anaerobic Chemoorganotrophs—Anaerobic Respiration tors of fecal pollution.
Desulfovibrio species reduce sulfur compounds to form hydrogen sulfide.
ECOPHYSIOLOGICAL DIVERSITY (table 11.2)
Anaerobic Chemoorganotrophs—Fermentation
11.6 ■ Thriving in Terrestrial Environments
Clostridium species form endospores. The lactic acid bacteria produce
lactic acid as their primary fermentation end product (figures 11.2, 11.3). Bacteria That Form a Resting Stage
Propionibacterium species produce propionic acid as their primary Bacillus and Clostridium species form endospores, the most resistant
fermentation end product. dormant form known (figure  11.14). Azotobacter species form cysts.
(figure 11.15). Myxobacteria aggregate to form fruiting bodies; within a
11.2 ■ Anoxygenic Phototrophs
fruiting body, cells form dormant microcysts (figure 11.16). Streptomyces
The Purple Bacteria species form chains of conidia at the end of hyphae. Many species natu-
The purple bacteria appear red, orange, or purple; the components of rally produce antibiotics (figure 11.17).
their photosynthetic apparatus are all within the cytoplasmic membrane. Bacteria That Associate with Plants
The Green Bacteria Agrobacterium species cause plant tumors. They transfer a portion of the
The green bacteria are typically green or brownish. Their accessory Ti plasmid to plant cells, genetically engineering the plant cells to pro-
pigments are often located in chlorosomes. duce opines and plant growth hormones. Rhizobia reside as endosym-
bionts in nodules on the roots of legumes, fixing nitrogen (figure 11.18).
Other Anoxygenic Phototrophs
Other anoxygenic phototrophs have been discovered, including some 11.7 ■ Thriving in Aquatic Environments
that form endospores.
Sheathed Bacteria
11.3 ■ Oxygenic Phototrophs Sheathed bacteria attach to solid objects in favorable habitats; the sheath
shelters them from attack by predators (figure 11.20).
The Cyanobacteria (figures 11.6, 11.7)
Genetic evidence indicates that chloroplasts evolved from a species of Prosthecate Bacteria
cyanobacteria. Nitrogen-fixing cyanobacteria are critically important Caulobacter species have a single polar prostheca called a stalk; at
ecologically, because they provide an available source of both carbon the tip of the stalk is a holdfast. The cells divide by binary fission
and nitrogen. Filamentous species maintain the structure and productiv- (figure 11.21). Hyphomicrobium species divide by forming a bud at the
ity of some soils. Some species of cyanobacteria produce toxins that can tip of their single polar prostheca (figure 11.22).
be deadly to animals that ingest contaminated water.
Bacteria That Derive Nutrients from Other Organisms
11.4 ■ Aerobic Chemolithotrophs Bdellovibrio species prey on other bacteria (figure 11.23). Certain species
of bioluminescent bacteria form symbiotic relationships with specific
The Sulfur-Oxidizing Bacteria
types of squid and fish (figure 11.24). Epulopiscium species reside within
The filamentous sulfur oxidizers Beggiatoa and Thiothrix live in sulfur the intestinal tract of surgeonfish. Legionella species often reside within
springs, in sewage-polluted waters, and on the surface of marine and protozoa; they can cause respiratory disease when inhaled.
freshwater sediments (figure 11.9). Acidithiobacillus species are found in
both terrestrial and aquatic habitats (figure 11.10). Bacteria That Move by Unusual Mechanisms
Spirochetes move by means of endoflagella (figure  11.25). Magnetotactic
The Nitrifiers
bacteria contain a string of magnetic crystals that allow them to move up
Ammonia oxidizers convert ammonia to nitrite; they include or down in water or sediments to the microaerophilic niches they require.
Nitrosomonas and Nitrosococcus. Nitrite oxidizers convert nitrite to
nitrate; they include Nitrobacter and Nitrococcus. Bacteria That Form Storage Granules
Spirillum volutans forms polyphosphate granules. Thioploca species
The Hydrogen-Oxidizing Bacteria
“commute” to nitrate-rich waters. Thiomargarita namibiensis, the larg-
Aquifex and Hydrogenobacter species are thought to be among the est bacterium known, stores sulfur and has a nitrate-containing vacuole.
earliest bacterial forms to exist on earth.
11.8 ■ Animals as Habitats (table 11.3)
11.5 ■ Aerobic Chemoorganotrophs
Bacteria That Inhabit the Skin
Obligate Aerobes
Staphylococcus species are facultative anaerobes.
Micrococcus species are found in soil and on dust particles, inanimate
objects, and skin (figure 11.11). Mycobacterium species are widespread Bacteria That Inhabit Mucous Membranes
in nature. They are acid-fast. Pseudomonas species are widespread in Bacteroides species inhabit the mouth, intestinal tract, and genital tract
nature and have extremely diverse metabolic capabilities (figure 11.12). of humans and other animals. Bifidobacterium species reside in the
Thermus aquaticus is the source of Taq polymerase, a component of intestinal tract of animals, including humans, particularly breast-fed
 Part II The Microbial World 281

infants. Campylobacter and Helicobacter species are microaerophilic. mammals, it resides within the filarial worms that cause river blind-
Haemophilus species require compounds found in blood. Neisseria ness and elephantiasis.
species are nutritionally fastidious aerobes that grow in the oral cavity
11.9 ■ Archaea That Thrive in Extreme Conditions
and genital tract. Mycoplasma species lack a cell wall; they often have
sterols in their membrane that provide strength and rigidity (figure 11.26). Extreme Halophiles
Treponema and Borrelia species are spirochetes that typically inhabit Extreme halophiles are found in salt lakes, soda lakes, and brines used
mucous membranes and body fluids of humans and other animals. for curing fish (figure 11.29).
Obligate Intracellular Parasites Extreme Thermophiles
Species of Rickettsia, Orientia, and Ehrlichia are spread when a flea Methanothermus species are hyperthermophiles that make methane.
or tick transfers bacteria during a blood meal. Coxiella burnetii sur- Sulfur-reducing hyperthermophiles are obligate anaerobes that use
vives well outside the host due to sporelike structures (figure  11.27). sulfur as a terminal electron acceptor. Nanoarchaea grow as spheres
Chlamydia and Chlamydophila species are transmitted directly from attached to Ignicoccus species. Sulfur-oxidizing hyperthermophiles
person to person (figure  11.28). Wolbachia pipientis alters the repro- use O2 as a terminal electron acceptor, generating sulfuric acid.
ductive biology of infected arthropods; although it does not infect Thermophilic extreme acidophiles have an optimum pH of 2 or below.

Review Questions
Short Answer 6. Which of the following genera preys on other bacteria?
1. What kind of bacteria might compose the subsurface scum of pol- a) Bdellovibrio b) Caulobacter c) Hyphomicrobium
luted ponds? d) Photobacterium e) Sphaerotilus
2. What kind of bacterium might be responsible for plugging the 7. All of the following genera are obligate intracellular parasites
pipes in a sewage treatment facility? except
3. Give three examples of energy sources used by chemolithotrophs. a) Chlamydia. b) Coxiella. c) Ehrlichia.
4. Name two genera of endospore-forming bacteria. How do they d) Mycoplasma. e) Rickettsia.
differ? 8. Which of the following genera are known to fix nitrogen?
5. How is the life cycle of Epulopiscium species unusual? 1. Anabaena 2. Azotobacter 3. Deinococcus
6. What unique motility structure characterizes the spirochetes? 4. Mycoplasma 5. Rhizobium
7. In what way does the metabolism of Streptococcus species differ a) 1, 3, 4 b) 1, 2, 5 c) 2, 3, 5
from that of Staphylococcus species?
d) 2, 4, 5 e) 3, 4, 5
8. How have species of Streptomyces contributed to the treatment of
9. Which of the following archaea would most likely be found coex-
infectious diseases?
isting with bacteria?
9. What characteristics of Azotobacter species protect their nitroge-
a) Nanoarchaeum b) Halobacterium c) Methanococcus
nase enzyme from inactivation by O2?
d) Picrophilus e) Sulfolobus
10. Compare and contrast the relationships of Agrobacterium and
Rhizobium species with plants. 10. Thermoplasma and Picrophilus grow best in which of the follow-
ing extreme conditions?
Multiple Choice a) Low pH b) High salt c) High temperature
1. A catalase-negative colony growing on a plate that was incubated d) a and c e) b and c
aerobically could be which of these genera?
a) Bacillus b) Escherichia c) Micrococcus
Applications
d) Staphylococcus e) Streptococcus 1. A student argues that it makes no sense to be concerned about coli-
forms in drinking water because they are harmless members of our
2. All of the following genera are spirochetes except
normal microbiota. Explain why regulatory agencies are concerned
a) Borrelia. b) Caulobacter. c) Leptospira. about coliforms.
d) Spirochaeta. e) Treponema. 2. A friend who has lakefront property and cherishes her lush green lawn
3. Which of the following genera would you most likely find growing complains of the green foul-smelling scum on the lake each summer.
in acidic runoff from a coal mine? Explain how her lawn might be contributing to the problem.
a) Clostridium b) Escherichia c) Lactic acid bacteria
d) Thermus e) Acidithiobacillus Critical Thinking +
4. The dormant forms of which of the following genera are the most 1. Soil often goes through periods of extreme dryness and extreme
resistant to environmental extremes? wetness. What characteristics of Clostridium species make them
well suited for these conditions?
1. Azotobacter 2. Bacillus 3. Clostridium
2. Some organisms use sulfur as an electron donor (a source of energy),
4. Myxobacteria 5. Streptomyces
whereas others use sulfur as an electron acceptor. How can this be if
a) 1, 2 b) 2, 3 c) 3, 4 there must be a difference between the electron affinity of electron
d) 4, 5 e) 1, 5 donors and acceptors for an organism to obtain energy?
5. Members of which of the following genera are coliforms?
a) Bacteroides b) Bifidobacterium c) Clostridium
d) Escherichia e) Streptococcus
12
The Eukaryotic Members
of the Microbial World
KEY TERMS
Algae Photosynthetic protists. an immature form of a parasite
occurs.
Arthropod Type of animal such as
an insect or arachnid. Mycosis Disease caused by fungal
infection.
Definitive Host Organism in
which sexual reproduction or the Phytoplankton Microscopic free-
adult form of a parasite occurs. floating photosynthetic organisms.
Fungus Non-photosynthetic Protists Eukaryotes that are not
eukaryotic organism with a chitinous fungi, plants, or animals.
cell wall. Protozoa Heterotrophic protists.
Helminth A parasitic worm. Yeasts Unicellular fungi that are
Intermediate Host Organism not chytrids.
in which asexual reproduction or

chemicals and resistance genes. The battle continues as late blight results
in global losses of $10 billion per year, causing devastation in developing
countries where many still rely on potatoes as their main source of nutrition.

A
Diatoms, a type of algae, have complex cell walls.
ll eukaryotic organisms are in the domain Eucarya, a diverse
group ranging from microscopic members to plants and
animals. Microscopic eukaryotes include fungi, algae, pro-
A Glimpse of History tozoa, slime molds, and water molds. In this chapter, we also include
Sometimes, a single event can change the course of history. A water multicellular worms and certain insects. Although usually visible to
mold, Phytophthora infestans, which causes a disease called late blight the naked eye, many are implicated in human disease. Moreover, they
in potatoes, triggered just such an event—the Irish Potato Famine. From are often carried or transmitted in forms that are indeed microscopic.
1845 to 1847, the potato crop in Ireland was decimated by the disease, and Eucarya, p. 10 domains, p. 238
the entire population of that nation faced starvation. Estimates vary, but it Eukaryotic organisms have traditionally been classified on
is likely that 1.5 million people died during the famine, while more than the basis of gross anatomical characteristics. Such classification
1 million emigrated to other countries—primarily the United States and has always been problematic, however, because it does not neces-
Canada. The population of Ireland fell by about 25% during this period.
sarily reflect evolutionary relationships. Modern DNA sequencing
Two hundred years earlier, the Spaniards brought potatoes to Europe
from South America. The potato became the main source of nutrition for
makes it possible to examine these organisms at the molecular
many Irish people because it was easy to grow and provided a convenient level, but there are still many problems in developing an accu-
and nearly complete food source that could be stored for months. Irish rate classification scheme. Because of this, some microscopic
dependence on this single food source left them open to a major disaster eukaryotes are still discussed in informal groups that do not reflect
when disease destroyed that food source. their evolutionary relatedness. For example, the term algae is
Phytophthora infestans affects every part of the potato plant includ- used to collectively refer to simple autotrophic (photosynthetic)
ing the leaves, stems, and tubers (potatoes). The potato turns to a black, eukaryotes, fungi are heterotrophic organisms that have chitin in
mushy mess. Once a field is contaminated, it is very difficult to rid it their cell wall, and protozoa are microscopic heterotrophic organ-
of the infectious agent. Because the tuber is underground, an infection isms that are not fungi. Protists are eukaryotes that are not fungi,
is often not noticed until it has destroyed the entire plant. The infection plants, or animals. photosynthesis, p. 151
is transmitted by spores, which can persist for years in the soil and are
Eukaryotes are characterized by a membrane-bound nucleus
spread by wind as well as by those who handle the potatoes.
Plant breeders and scientists still battle potato blight. In 2009, the
(eukaryote means “true nucleus”). This organelle contains their genetic
potato genome was sequenced, and scientists hope to locate resistance information, packaged in structures called chromosomes. Because
genes that can be promoted in crop potatoes. The same year the genome their DNA is located in the nucleus and the ribosomes are in the
of Phytophthora infestans was also sequenced. Long expanses of repeti- cytoplasm, the processes of transcription and translation cannot occur
tive genes coding for enzymes that attack the plants were located in the simultaneously, as it does in prokaryotic cells. Eukaryotes also have
pathogen, providing ready sources of variation that can evade the action of lysosomes, Golgi, and other membrane-bound organelles that allow
282
 Part II The Microbial World 283

They reproduce by binary fission: The chromosome is replicated,

and then the cell elongates and divides, with each daughter cell
receiving a copy of the chromosome (see figure 4.1). Eukaryotes
Haploid may be haploid or diploid and often progress through a complex
cells (n) Mitosis Mitosis life cycle involving both forms. They may reproduce asexually by
a type of nuclear division called mitosis, in which each daughter
Haploid organism (n) Sex cells (n) cell receives exactly the same chromosome complement as the
parent cell, whether the parent cell is haploid or diploid. They
may also reproduce using meiosis, in which diploid cells produce
n
haploid cells (often called spores) that can develop into haploid
Meiosis Meiosis Fusion of organisms or directly into haploid sex cells called gametes that
2n sex cells can be used in sexual reproduction (figure 12.1). Gametes are
often flagellated and highly mobile. Fusion of two gametes forms
Diploid a diploid cell, allowing recombination of genetic material. This
organism (2n) sexual process increases genetic variation that can be used as the
raw material for natural selection, the basis of evolution.
Mitosis
MicroByte
Eukaryotic pathogens can be difficult to target with medication
because their cell components are often the same as those of humans.
Diploid cell (2n)

FIGURE 12.1 Cell Division Meiosis gives rise to haploid cells that 12.1 ■ Fungi
can grow into a haploid organism by mitosis or can be used as sex
cells (gametes) in sexual reproduction. Fusion of gametes forms a
Learning Outcomes
diploid cell that can grow into a diploid organism by mitosis or can
undergo meiosis to form haploid cells. 1. Describe the structure, habitat, and reproductive strategies of a
yeast and a mold.
? Can a gamete be diploid?
2. Explain the important symbiotic relationships of fungi.
3. Name and describe the roles of economically and medically
compartmentalization of cell processes. Energy transformation in important fungi.
eukaryotes primarily occurs in membrane-rich mitochondria and
chloroplasts where electron transport chains operate; in prokaryotes, The study of fungi is known as mycology. Fungi include molds,
these are associated with the plasma membrane. Many eukaryotes single-celled yeasts, and the familiar mushroom. These terms have
do not have a cell wall, and when they do, it lacks peptidoglycan, the nothing to do with the classification of fungi, but instead refer to
molecule that characterizes the prokaryotic cell wall. Finally, most their morphological forms (figure 12.2).
eukaryotic cells have a well-developed cytoskeleton that is important
■ Yeasts are single-celled fungi.
in movement and maintenance of structure (see figure 3.49) organ-
elles, p. 68 transcription, p. 168 ■ Molds are filamentous fungi.
The mode of reproduction in eukaryotes is fundamentally dif- ■ Mushrooms are simply the reproductive structures of certain
ferent from that of prokaryotes. Recall that prokaryotes are typi- fungi, similar to a peach on a peach tree. Like peaches, some
cally haploid, meaning they carry only one copy of the genome. large mushrooms are edible.

Mycelium
FIGURE 12.2
Morphology of Fungi
(a) Microscopic Candida
albicans; the circles are
asexual reproductive spores.
(b) The cottony white mass
inside the potato is a mold.
(c) Amanita muscaria, a
highly poisonous mushroom.
? What chemical
characterizes the cell
wall in each of these
fungal forms?

(a) (b) (c)

284 Chapter 12 The Eukaryotic Members of the Microbial World

Fungi are characterized by a cell wall that contains chitin— nature. We will describe only four major fungal groups: chytrids
the same molecule found in the exoskeleton of insects. It is some- (Chytridiomycota), zygomycetes (Zygomycota), ascomycetes
what stronger than the cellulose-based cell wall of plants, and (Ascomycota), and basidiomycetes (Basidiomycota) (table 12.1).
chemically distinct from the peptidoglycan cell wall of bacteria. The real diversity within fungi is much more complex.
Fungal membranes typically have ergosterol, distinguishing them The chytrids usually live in water, but some live in the guts
from animal cell membranes, which have cholesterol. Ergosterol of mammalian herbivores where they help with the digestion of
is the target for many antifungal medications. plant material. Some are parasitic. For example, Batrachochytrium
To obtain nutrients, fungi secrete enzymes into the environ- dendrobatidis can infect the skin of frogs and is believed to be
ment to break large molecules into smaller ones that they can responsible for the catastrophic decline in frog populations over
absorb. Along with bacteria, fungi are the principal decomposers the past decade. Chytrids are the only type of fungus with motile
of organic compounds because they can degrade both cellulose forms; the reproductive cells have flagella.
and lignin, the main components of wood. This decomposition The zygomycetes include the common black bread mold,
releases carbon dioxide into the atmosphere and nitrogen com- Rhizopus, and other organisms that often spoil fruits and vegeta-
pounds into the soil, which are then taken up by plants and again bles. The black growth that you see on bread is actually reproduc-
converted into organic compounds. Without this recycling of tive structures called sporangia that support and house hundreds of
organic material, the earth would quickly be overrun with organic asexual spores called sporangiospores (figure 12.3).
waste. biogeochemical cycles, p. 725 The ascomycetes (sac fungi) are a diverse group with about
In their key role as decomposers, fungi are saprophytic, 75% of all known fungi. They include Penicillium, the source of
absorbing nutrients from dead or decaying organic matter. Some the earliest recognized antibiotic—penicillin. Other ascomycetes
fungi, however, can absorb nutrients from living tissue, acting as are pathogens such as those that cause Dutch elm disease. Some
parasites. Relatively few fungi infect humans—although athlete’s ascomycetes are highly prized for the flavor of their reproductive
foot and vaginal yeast infections are not uncommon—but many structures (morels and truffles). Many ascomycetes form lichens
plant species suffer devastating fungal infections. More often, in association with a photosynthetic partner.
fungi partner with other organisms in mutually beneficial relation- The basidiomycetes (club fungi) are commonly recognized
ships. For example, fungi and algae form lichens that can grow by their reproductive structures—mushrooms—that are often col-
on surfaces where neither can survive alone. parasitism, p.  381 lected or grown for food. The group also includes plant parasites
mutualism, p. 381 like rusts and smuts, which earn their common names because of
their appearance on the infected plant. Significant losses in wheat,
Types of Fungi rye, and corn crops have been caused by these fungi.
Classification of fungi is in a state of flux and will continue
to be so until the genomes of many more species have been Structure of Fungi
sequenced. About 80,000 fungal species are recognized, but gene Most fungi are multicellular, composed of threadlike filaments
studies indicate that there are likely over 1 million species in called hyphae (figure 12.4). A visible mass of hyphae is a

TABLE 12.1 Characteristics of Major Groups of Fungi

Group and Usual Some Distinguishing Asexual Sexual
Representative Member(s) Appearance Habitat Characteristics Reproduction Reproduction
Chytridiomycetes Aquatic, Unicellular and Motile zoospores Flagellated gametes
Batrachochytrium dendrobatidis guts of multicellular. Rhizoids from a sporangium in male and female
herbivores, with no true mycelium
parasitic

Zygomycetes Terrestrial Multicellular, mycelia of Asexual spores Sexual spores known

Rhizopus stolonifer continuous hyphae with develop in sporangia as zygospores can
(black bread mold) many haploid nuclei on the tips of aerial remain dormant
hyphae in adverse
environment

Ascomycetes Terrestrial, Unicellular and Is common Involves the

Neurospora, Saccharomyces on fruit multicellular with by budding; formation of an
cerevisiae (baker’s yeast) and other septated mycelia conidiospores ascus (sac) on
Penicillium, Aspergillus organic specialized hyphae
materials
Basidiomycetes Terrestrial Multicellular, uninucleated Commonly absent Produce
Agaricus campestris mycelia; group includes basidiospores that are
(meadow mushroom) mushrooms, smuts, rusts borne on club-shaped
Cryptococcus neoformans that affect the food structures at the tips
supply of the hyphae
 Part II The Microbial World 285

Exposure to such chronic levels of mold or spores can lead to

allergies or other health problems.
Parasitic fungi have specialized hyphae called haustoria that
protrude into host cells to gain nutrients. Haustoria do not pen-
etrate the host cell’s plasma membrane, but are surrounded by it,
Hyphae much like a hand in a mitten. Saprophytic fungi sometimes have
specialized hyphae called rhizoids, which anchor them to the sub-
strate. Fungal rhizoids do not function in exchange of materials.
Dimorphic fungi can grow as single yeast cells or multicellular
mycelia, depending on the environmental conditions. Some of these
fungi cause systemic disease in humans. For example, Histoplasma
capsulatum grows in the soil as a mold, and its reproductive spores
easily become airborne. When they are inhaled into the warm, moist
environment of the lungs, they develop into the yeast form, which
Sporangium can cause disease. histoplasmosis, p. 515
FIGURE 12.3 Black Bread Mold Rhizopus stolonifer, showing
MicroByte
threadlike reproductive hyphae (singular: hypha) and black sporangia
housing sporangiospores.
The largest organism on earth, called the humongous fungus, has a
mycelium that spans over 2,200 acres in Oregon.
? What is a spore?

mycelium (plural: mycelia). Fungi are generally not motile, so Fungal Habitats
they cannot move toward a food source. Instead, the tips of the Fungi are found in nearly every habitat on earth, including the
hyphae respond to a nutrient source by growing very quickly in thermal pools at Yellowstone National Park, volcanic craters, and
that direction. Cells in the hyphae undergo mitosis, sometimes lakes with very high salt content, such as the Great Salt Lake and
without accompanying cell division, resulting in a structure with the Dead Sea. They are mainly terrestrial organisms. Some fungal
many nuclei. Even when the cytoplasm divides after mitosis, species occur only on a particular strain of one genus of plants.
openings remain between cells that allow cytoplasm and organ- Others are widespread because they are extremely versatile in
elles to move along the length of the hypha. what they can degrade and use as a source of carbon and energy.
The high surface-to-volume ratio of hyphae makes them well As a group, fungi can degrade leather, cork, hair, wax, ink, jet
adapted to absorb nutrients. The enzymes that they release not fuel, carpet, drywall, and even some synthetic plastics.
only break down nutrient molecules; they also repel the growth of Some fungal species can grow in concentrations of salts, sug-
other hyphae. As a result, hyphae branch as they undergo mitosis ars, or acids strong enough to kill most bacteria. Because of this,
and spread throughout the food source, ensuring that each hypha fungi are often responsible for spoiling pickles, fruit preserves,
will have maximum access to nutrients. When you see mold and other foods. Different fungi can grow from pH as low as 2.2 to
growing on a food, you typically see only the hyphae involved in as high as 9.6, and they usually grow well at a pH of 5.0 or lower.
reproduction, often bearing reproductive spores. Within the food They can grow better than bacteria on acidic fruits and vegetables.
are the bulk of the hyphae that act in nutrient absorption. food spoilage, p. 756
Fungi are most successful in moist environments. Mildew Most fungi prefer an environment from 20°C to 35°C, but
and mold become problems, for example, in a damp basement. they can easily survive at refrigerator temperatures and below.

Mycelium

Spore Hypha

FIGURE 12.4 Formation of Hyphae and Mycelium The white mass of hyphae form a tangled mass called the
mycelium. The mycelium forms when spores of fungi germinate and then elongate to form filaments called hyphae
that intertwine.
? Where do fungi digest their food?
286 Chapter 12 The Eukaryotic Members of the Microbial World

FIGURE 12.5 Lichens (a) A lichen consists of cells

of an autotroph, either an alga or a cyanobacterium, Alga
entwined within the hyphae of the fungal partner.
(b) Lichens on a fallen tree.
? What does the fungus contribute
to the lichen?

Fungus

Fungal
hyphae Cortex

Algal
layer

Cortex

(a) (b)

Some are resistant to pasteurization, and others can grow at tem- Some fungi grow in a mutually beneficial association with
peratures below the freezing point of water. pasteurization, p. 112 plant roots, forming mycorrhizas (figure 12.6). The high surface
Most fungi are aerobic, but some of the yeasts are facultative area of fungal hyphae increases the plant’s ability to absorb water
anaerobes, producing ethanol by fermentation. Some obligate and minerals. The fungus also supplies the plant with nitrogen and
anaerobes live in the rumen of cows, where they are important in phosphorus from the breakdown of organic material in the soil.
the digestion of the plant material. obligate anaerobe, p. 90 The plant, in return, supplies the fungus with organic compounds.
It is estimated that 80% of vascular plants have some type of
mycorrhizal association. Plants with mycorrhizas grow better than
Symbiotic Relationships of Fungi those without. Orchids cannot grow without mycorrhizas that help
Lichens result from the association of a fungus with a photosynthetic provide nutrients to the young plant. mycorrhizas, p. 730
organism such as an alga or a cyanobacterium (figure 12.5). The Certain insects also depend on symbiotic relationships
fungus provides protection and absorbs water and minerals for the with fungi. For example, leaf-cutting ants farm their own fun-
pair. The photosynthetic member supplies the fungus with organic gal gardens (figure 12.7). The ants cannot eat tropical vegeta-
nutrients. Lichens can grow in extreme ecosystems where neither tion because the leaves are often poisonous. Instead, the ants
partner could survive alone. For example, they grow in sub-Arctic chop the plant leaves into bits and add a mycelium. The fungi
tundra where they are the primary diet of reindeer. Lichens are grow, secreting enzymes that digest the plant material, and
often the first organisms to grow on bare rock where they can eventually produce reproductive structures that the ant then
begin the process of soil formation. In spite of their hardiness, uses as its food source.
lichens are often a good indicator of air qual-
ity. Polluted air is lethal to lichens because they
absorb, but are unable to excrete, its common Reproduction in Fungi
components, including toxic metals, sulfur The reproductive structures of fungi are very important in identi-
dioxide, and ozone. You will fication. From a practical standpoint, the most important of these
not find many lichens are the forms that develop asexually, because they can be seen in
in industrial cities.

FIGURE 12.6 Mycorrhizas Fungi form intimate relationships with FIGURE 12.7 Leaf-Cutter Ants These ants carry food for fungi,
the roots of most green plants. They supply the plant with nitrogen whose reproductive structures then serve as a food source for the ants.
and phosphorus and increase the plant’s ability to absorb water.
? What type of fungal reproduction occurs when the ants place a
? Are mycorrhizas considered parasites? piece of mycelium on the bits of leaves?
 Part II The Microbial World 287

Gills FIGURE 12.8 Mushrooms (a) The extensive under-

ground mycelium gives rise to fruiting bodies called
mushrooms, composed of dense hyphae protecting the
spores produced under the gills. (b) Lepiota rachodes,
showing the fruiting bodies and gills.
? What group of fungi produces mushrooms?

6 cm
Gills
Hypha

Mycelium Mycelium
(a) (b)

pure cultures grown in the laboratory. However, the sexual forms

play an important role in fungal classification. The sexual forms A
of some fungi have never been seen, which made it difficult to
classify these organisms before DNA sequencing methods were 1
developed. Further complicating matters, many fungi have been
inadvertently “discovered” twice—once based on the sexual Nucleus
reproductive forms and once based on the asexual forms—so they
are known by two different names!
The reproductive cells of a fungus come in a variety of sizes Cell wall softens at point A.
and shapes. Some mycologists use the term spore to refer to
reproductive cells that are formed either sexually or asexually. Bud
Asexual spores, however, are more typically called conidia, or in
zygomycetes, sporangiospores. Reproductive cells may be housed 2
in special structures called sporangia in zygomycetes or asci Microtubules
(singular: ascus) in ascomycetes. They may simply be exposed on
the tips of reproductive hyphae. Even when exposed, they may be
somewhat protected in basidiomycetes beneath the dense hyphae
of a mushroom or puffball (figure 12.8). Only chytrids produce Nucleus divides by mitosis.
motile gametes.
Since most fungal reproductive cells are not motile, sexual
reproduction results from fusion of hyphae from two different
mating types growing toward one another. Often after this fusion, 3

the cells of the fungus will house both haploid nuclei, forming a
distinct fungal structure called a dikaryon (meaning two nuclei).
Eventually, the nuclei fuse and then undergo meiosis, forming
haploid spores. Because the two mating types have no obvious
differences, mycologists refer to them as “+” and “–”. One nucleus migrates into the bud.
Some asexual reproduction does not involve spores. Yeast
cells, for example, may simply reproduce by mitosis. Yeasts may
also reproduce by budding, in which daughter cells pinch off from
4
a larger parent cell, often leaving a scar on the larger cell, which
eventually dies after producing a number of buds (figure 12.9).
Molds may reproduce asexually by fragmentation. In this type New cell
of reproduction, a portion of the parent organism breaks off and
grows into a new organism, and the parent organism survives.
Hyphae may grow from a piece of a mycelium placed on a suitable The bud breaks off.
nutrient source, as in the farming technique of leaf-cutter ants.
Fungal spores and conidia are small and numerous and easily FIGURE 12.9 Budding in Yeast The nucleus divides by
carried by wind or water. They provide some protection against mitosis and one nucleus moves into the bud. When the cell wall
drying and lack of nutrients and can persist for years until condi- grows together the bud breaks off, forming a new cell.
tions improve. When a fungal spore germinates, it begins to grow ? How is budding different from mitosis?
288 Chapter 12 The Eukaryotic Members of the Microbial World

hyphae in the direction of a food source. When the food source Some Medically Important
diminishes, spore formation typically increases. Note that fungal TABLE 12.2 Fungal Diseases
spores are quite different from bacterial endospores, which are
much more resistant to environmental conditions and are not a
means of reproduction.  endospore, p. 67 Page for More
Disease Causative Agent Information

Economic Importance of Fungi Candidial Candida albicans p. 545

skin infection
Many fungi are important commercially. They synthesize important
Coccidioidomycosis Coccidioides immitis p. 514
antimicrobial medicines such as penicillin and griseofulvin. Fungi
Cryptococcal Cryptococcus p. 660
are useful tools for studying complex eukaryotic events—such as
meningoencephalitis neoformans
cancer and aging—within a simple cell. The first eukaryotic genome
Histoplasmosis Histoplasma p. 515
sequenced was that of the yeast Saccharomyces cerevisiae, a model
capsulatum
organism used in a variety of genetic and biochemical studies. Yeasts
Pneumocystis Pneumocystis p. 708
have been genetically engineered to produce numerous important
pneumonia jiroveci
molecules such as human insulin and a vaccine against hepatitis B.
Sporotrichosis Sporothrix schenckii p. 565
 penicillin, p. 463  genetic engineering, p. 216

Saccharomyces cerevisiae, also known as brewer’s yeast or Vulvovaginal Candida albicans p. 618
candidiasis
baker’s yeast, has long been used in the production of wine, beer,
and bread. Other fungal species are useful in making the large Tinea versicolor Malassezia furfur p. 544
variety of cheeses produced throughout the world. Ironically,
fungi are also among the greatest spoilers of food products; tons of
food are discarded each year because they have been made ined- may also refer to the part of the body affected. Cutaneous mycoses
ible by fungal infections. Saccharomyces, p. 753 affect the hair, skin, or nails (see figure 22.26). They are caused
Fungi cause a number of crop diseases that cost billions of dol- by skin-invading molds called dermatophytes. Systemic mycoses
lars from expenditures on preventative measures or losses due to crop affect tissues deep within the body and are usually caused by inha-
damage. Grain crops, a large portion of our food supply, are particu- lation of spores. dermatophytes, p. 543
larly vulnerable to fungal infection. Dutch elm disease, caused by the Some fungi produce toxins. Aflatoxins, produced by
fungus Ophiostoma ulmi (Ulmus is a genus of elm), dramatically Aspergillus species, are the most studied of these toxins and are
changed the landscape of many U.S. cities when all of the elm trees considered carcinogenic. The U.S. Food and Drug Administration
lining streets and surrounding public buildings were killed. The cost (FDA) monitors levels of aflatoxins in foods such as grains and
of removing these diseased and dying trees was significant. peanuts. The rye mold Claviceps purpurea, also known as ergot,
produces a toxin with hallucinogenic properties. Some believe that
some young women in Salem, Massachusetts, in the late 1600s ate
Medical Importance of Fungi contaminated rye, and their resultant strange behavior led to accu-
Because relatively few fungal species infect humans, and many sations of witchcraft. The active chemical has been purified from
fungi produce important antimicrobial medications, their impact C. purpurea to yield the drug ergotamine, which decreases blood
on human health is probably a net positive. Still, some fungal flow. It is used to control uterine bleeding and relieve migraine
diseases are relatively common, such as athlete’s foot and jock headaches. Some fungi, such as Amanita species, produce power-
itch. Life-threatening fungal infections such as cryptococcal men- ful toxins that can cause fatal liver damage (see figure 12.2c).
ingitis are rare. An exception is in immunocompromised patients,
where fungal diseases are much more common and devastating. MicroAssessment 12.1
Cryptococcus, p. 660
Fungi have chitin in their cell walls and absorb nutrients after
Fungi cause human illnesses in three general ways: (1) a per- secreting digestive enzymes onto a food source. As saprophytes,
son develops an allergic or asthmatic reaction to the fungus or its they are important recyclers of carbon and other elements. Fungi
spores; (2) the fungus grows on or in the human body, causing a can also form important relationships with other organisms, such as
fungal disease, or mycosis, and (3) the fungus produces toxins that seen in lichens or mycorrhizas. Fungi may be single-celled yeasts
a person ingests. Table 12.2 lists some fungal diseases and serves or multicellular molds that grow by extending tubelike hyphae
into a food source. They may reproduce sexually by fusion of
as a directory to where they are discussed in upcoming chapters.
hyphae of different mating types, or asexually by spores, budding,
Fungal spores or conidia are found everywhere on earth up or fragmentation. Fungi cause significant plant diseases and food
to altitudes of more than 7 miles. The air we breathe may contain spoilage but also are a food source and are used in food production.
more than 10,000 of these cells per cubic meter. People with aller- Relatively few fungi cause human disease.
gies often monitor published pollen and mold counts and avoid 1. How can fungi cause disease in humans?
unnecessary outdoor activity when levels are high. Fungal spores
2. Describe fungal symbiotic relationships with algae, green plants, and
can also trigger asthma attacks. asthma, p. 404 insects, and explain how the relationships benefit each partner.
The names of mycoses often reflect the causative agent. For
3. Why would spore formation increase when food supplies are
example, diseases caused by the yeast Candida albicans are called diminishing? +
candidiasis and are among the most common mycoses. Infections
 Part II The Microbial World 289

12.2 ■ Algae Apicomplexans

Dinoflagellates
Learning Outcomes
4. Explain how various types of algae differ from one another and
from other members of the eukaryotic microbial world.
5. Describe how algae can affect human health. Brown algae

The term algae refers to simple photosynthetic eukaryotes. Chromalveolates Diatoms

Algae differ from other eukaryotic photosynthetic organisms
such as land plants because they lack an organized vascular Oomycetes (water molds)
system and they have relatively simple reproductive structures.
Most algae are aquatic; they may be microscopic or macro-
scopic. Algae do not directly infect humans, but some produce
toxins that may be concentrated in other animals that humans
use for food. Plantae
Red algae
Algae contain pigments that absorb radiant energy. That
energy is used in the process of photosynthesis, converting
CO2 and H2O to organic material and oxygen. Algae are Green algae (chlorophytes)
essential for maintaining life in aquatic environments because
other organisms depend upon the organic molecules for food. Land plants
photosynthesis, p. 151
Green algae (charophytes)

Types of Algae
Diplomonads
As noted earlier, algae are not a strict classification. They are a
diverse group of protists that share some fundamental charac- Parabasilids
teristics, but are not necessarily related. Figure 12.10 shows a Excavates
simple taxonomic representation of the protists in this chapter.
The various types of algae are often characterized by the
pigments they contain. All algae contain chlorophyll a, a pig-
ment also found in green plants and cyanobacteria. Red algae Heteroloboseans
and brown algae also contain other pigments that absorb dif-
Euglenids
ferent wavelengths of light and give them their characteristic
appearance. These pigments extend the range of light waves
that can be used for photosynthesis. Some of the general char- Kinetoplastids
acteristics of algal groups are summarized in table 12.3.

Structure of Algae Rhizaria Foraminiferans

Algae can be either unicellular or multicellular. All algae, how-
ever, contain chloroplasts with photosynthetic pigments and
have rigid cell walls mostly composed of cellulose. Some multi-
cellular species contain large amounts of other compounds in
their cell walls. Cell walls of red algae contain agar used by Fungi
scientists to solidify growth media; cell walls of brown algae
Animals
contain alginic acid used to provide the consistency in products
like ice cream and cosmetics. chloroplasts, p. 76 agar, p. 86

Microscopic Algae Unikonts

Microscopic algae can be single-celled organisms floating free

or propelled by flagella, or they can grow in long chains or
Loboseans

Plasmodial slime molds

FIGURE 12.10 A Phylogeny of Eukaryotes Based on Ribosomal RNA Sequence
Comparisons Algae are highlighted in green. Non-protists are shown in red. Note that not
all of the groups shown are at the same taxonomic level. Cellular slime molds

? Are red algae more closely related to green algae or to brown algae?
290 Chapter 12 The Eukaryotic Members of the Microbial World

TABLE 12.3 Characteristics of Major Groups of Algae

Principal
Pigments
(in addition to Storage Mode of
Group Usual Habitat chlorophyll a) Product Cell Wall Reproduction

Green algae Fresh water; salt Chlorophyll Starch Cellulose and Asexual by multiple
water; soil; tree b; carotenes; pectin fission; spores or sexual
bark; lichens xanthophylls
Brown algae Salt water Xanthophylls, Starchlike Cellulose and Asexual, motile
especially carbohydrates; pectin; alginic acid zoospores; sexual,
fucoxanthin mannitol; fats motile gametes
Red algae, Mostly salt water, Phycobilins; Starchlike Cellulose and Asexual spores; sexual
corallines several genera in carotenes; carbohydrates pectin; agar; gametes
fresh water xanthophylls carrageenan
Diatoms, golden Fresh water; Carotenes Starchlike Pectin, often Asexual or sexual
brown algae salt water; soil; carbohydrates impregnated with
higher plants silica or calcium
Dinoflagellates Mostly salt water Carotenes; Starch; oils Cellulose and Asexual; rarely sexual
but common in xanthophylls pectin
fresh water
Euglenids Fresh water Chlorophyll Fats; starchlike Lacking, but elastic Asexual only by binary
b; carotenes; carbohydrates pellicle present fission
xanthophylls

filaments. The unicellular algae—including diatoms, as well as to obtain water and nutrients for the organism. It functions simply to
some green algae, dinoflagellates and euglenids, and a few red anchor the organism to a firm substrate. The stalk of a multicellular
algae—are well adapted to an aquatic environment. As single alga, known as the stipe, usually has leaflike structures or blades
cells, they have relatively large, absorptive surfaces, thus effec- attached to it. The blades are the main site of photosynthesis, although
tively using the dilute nutrients available. Some microscopic algae some also bear reproductive structures. Many large algae have gas-
such as Volvox form colonies of 500 to 60,000 biflagellated cells, containing bladders that keep the blades floating on the surface of the
which can be visible to the naked eye (figure 12.11). water to maximize exposure to sunlight.
Diatoms are abundant in aquatic environments and are dis-
tinguished by silicon dioxide incorporated into their cell walls.
When these organisms die, they sink to the bottom of the ocean, Algal Habitats
and the cell wall does not readily decompose (see chapter-opening Algae are found in both fresh and salt water, as well as in moist
photo). Deposits of diatoms are mined for a substance known as soil. Since the oceans cover more than 70% of the earth’s surface,
diatomaceous earth, used for filtering systems, abrasives in polishes, aquatic algae are major producers of molecu-
and many other purposes. Diatoms store a type of oil as an energy lar oxygen as well as important users of
reserve that helps them float at a desired depth. Chemical and physical carbon dioxide. Algae must live at
changes to diatoms that sank to the ocean floor millions of years water levels that allow penetra-
ago eventually converted them to a major source of crude oil and tion of light. Algae with dif-
natural gas. ferent pigments can occupy
different levels and thus
Macroscopic Algae avoid competition.
Macroscopic algae include Small organisms that
multicellular brown algae, float or drift near the surface
green algae, and red algae of aquatic environments are
(figure 12.12). Some of these called plankton. Unicellular
possess a structure called a algae make up a significant
holdfast, which looks like a part of the phytoplankton
root system but it is not used (phyto means “plant” and plank-
ton means “drifting”), the free-floating,
photosynthetic organisms found in plankton.
FIGURE 12.11 Volvox A colony of cells forms a Phytoplankton forms the base of aquatic food chains with
hollow sphere. The yellow-green circles are reproduc- all other organisms dependent upon them. For example,
tive cells that will eventually become new colonies. microscopic heterotrophs in the zooplankton (zoo means
? What pigment gives the green color to Volvox? “animal”) graze on phytoplankton, and then both become
 Part II The Microbial World 291

FIGURE 12.12 Types of Algae (a) This young brown alga, Nereocystis luetkeana
(bladder kelp), has a large bladder filled with gas. The blades are the most active sites for
photosynthesis. The holdfast anchors the kelp to rocks or other surfaces. In a single season,
kelp can grow to lengths of 5 to 15 m. (b) Sea lettuce, a green alga. (c) Corallina gracilis,
Blade red coral algae.
? What is the function of the gas bladder in kelp?

Bladder

Stipe

Holdfast

(a) 0.3 m (b) (c)

food for other organisms including benthic whales, some of the world’s poisoning, including numbness, dizziness, muscle weakness, and
largest mammals. food chain, p. 720 impaired respiration. Death can result from respiratory failure.
Cooking the shellfish does not destroy these toxins. Gonyaulax spe-
MicroByte cies are found in both the North Atlantic and the North Pacific and
More O2 is produced by algae in the oceans than by all forests on have seriously affected the shellfish industry on both coasts.
earth combined.

MicroAssessment 12.2

Algal Reproduction Algae are a diverse group of aquatic eukaryotic organisms that contain
chlorophyll a and carry out photosynthesis. Algae form the base of
Some algae, especially multicellular filamentous species, repro- aquatic food chains and produce much of our atmospheric oxygen.
duce asexually by fragmentation. Many algae alternate between a Algae do not cause disease directly, but they can produce toxins that
haploid generation and a diploid generation. Sometimes, as is the are harmful when ingested by humans.
case with Ulva (sea lettuce), the generations look physically simi- 4. What primary characteristics distinguish algae from other
lar and can be distinguished only by microscopic examination. In organisms?
other cases, the two forms look quite different. 5. What harmful effects can algae have on humans?
6. Why do algae have a greater variety of photosynthetic pigments
than land plants? +
Medical Importance of Algae
Although algae do not directly cause disease in humans, some
do so indirectly. Instances of disease most often occur dur-
ing algal blooms, during which the numbers of phytoplankton 12.3 ■ Protozoa
increase dramatically due to changes in water conditions. An
upwelling of water due to warmer temperatures often brings Learning Outcomes
more nutrients from the ocean bottom to the surface. When 6. Describe methods of reproduction in protozoa.
organisms encounter warmer waters and additional nutrients, 7. Describe the major diseases of humans that are caused by protozoa
they multiply rapidly. The runoff of fertilizers along waterways and name the causative agents.
and coastlines, or pollution from untreated sewage, may also
cause algae to proliferate. Algal blooms of dinoflagellates are The term protozoa has little meaning today as we learn more
commonly known as red tides. about the evolutionary diversity of the eukaryotes through genetic
Dinoflagellates of Gonyaulax species produce neurotoxins such sequencing studies. Organisms classically considered protozoa
as saxitoxin and gonyautoxin, some of the most potent non-protein (“animal-like” unicellular organisms) form a diverse group
poisons known. Shellfish such as clams, mussels, scallops, and in which most members bear little relationship to the others.
oysters feed on these dinoflagellates without apparent harm Protozoa are most easily defined by what they are not. In this
and, in the process, accumulate the neurotoxin in their tissues. When chapter, protozoa are unicellular heterotrophic organisms that are
humans eat the shellfish, they suffer symptoms of paralytic shellfish not fungi, slime molds, or water molds.
292 Chapter 12 The Eukaryotic Members of the Microbial World
Plasmodium
Toxoplasma
Types of Protozoa
Cryptosporidium
Protozoa are protists, along with the algae, slime Apicomplexans
Cyclospora
molds, and water molds. As with algae, protozoa
Dinoflagellates
are not a unified group but appear along an evolu-
tionary continuum (figure 12.13). Protozoa have
traditionally been classified primarily based on their
mode of locomotion. Examination of ultrastructure
Brown algae
and DNA sequences coding for rRNA, however,
show that organisms may not be closely related just Chromalveolates Diatoms
because they all use flagella, cilia, or pseudopodia
for locomotion. Protozoans are extremely diverse, Oomycetes (water molds)
but we will concentrate on members of this group
that cause human disease.
Apicomplexans are parasites with a structure
called an apical complex at one end that helps them
to penetrate the cell membrane of host cells. Many Plantae
Red algae
have complex life cycles that alternate between
sexual and asexual forms. This group includes
Plasmodium species that cause malaria, making them Green algae (chlorophytes)
one of the most significant causes of infectious dis-
ease in the world. It also includes the human patho- Land plants
gens Toxoplasma gondii, Cryptosporidium parvum,
and Cyclospora cayetanensis. malaria, p. 686 Green algae
Diplomonads and parabasalids are groups of (charophytes)
flagellated protists that lack mitochondria. Diplo-
monads typically have two nuclei and reside within Diplomonads Giardia
hosts where conditions are anaerobic or in stagnant
Parabasilids Trichomonas
water where O2 levels are low. The group includes Excavates
Giardia lamblia, a common cause of diarrhea in
campers who drink water directly from streams or
lakes (see figure 24.22). Parabasalids live within a
host organism. Some, for example, live within ter- Heteroloboseans Naegleria
mites and digest cellulose for their hosts. Others,
such as Trichomonas vaginalis, may cause disease. Euglenids
Parabasalids have a unique structure called a hydro- Trypanosoma
genosome, which in the absence of true mito- Kinetoplastids
Leishmania
chondria, produces some ATP while generating
hydrogen. Both groups reproduce asexually; a few
parabasalids also reproduce sexually. giardiasis,
p. 602 trichomoniasis, p. 635 Rhizaria Foraminiferans
Kinetoplastids have at least one flagellum and
are characterized by a distinctive complex mass
of DNA in their large single mitochondrion. This
mitochondrial DNA can produce variations in RNA
that may account for the rapid changes in cell sur- Fungi
face molecules that allow some of these organisms
to evade the immune system. This group includes Animals
Leishmania major, the cause of leishmaniasis, a
disease transmitted by sand flies. It also includes
Trypanosoma cruzi, the cause of Chagas’ disease, Unikonts
which kills up to 50,000 people worldwide each
year. The most significant species of the group is
Loboseans Entamoeba

FIGURE 12.13 Classification of Eukaryotes Highlighting the Plasmodial slime molds

Protozoans Note that not all groups are at the same level of classification.
? What do the diplomonads and the parabasalids have in common? Cellular slime molds
 Part II The Microbial World 293

Trypanosoma brucei, which causes African sleeping sickness (see marine environments, protozoa make up part of the zooplankton.
figure 26.19). sleeping sickness, p. 662 On land, protozoa are abundant in soil as well as in or on plants
Loboseans and heteroloboseans have an ameboid (flexible) and animals. Specialized protozoan habitats include the guts of
body form, but they are only distantly related to one another and to termites, roaches, and ruminants such as cattle.
the slime molds discussed later in this chapter. Loboseans change Protozoa are an important part of the food chain. They eat
shape as they move by extending and retracting pseudopodia and bacteria and algae and, in turn, serve as food for larger species.
engulfing food particles by phagocytosis. Entamoeba histolytica The protozoa help maintain ecological balance by devouring vast
infects humans, causing diarrhea ranging from mild asymptomatic numbers of bacteria and algae. food chain, p. 720
disease to severe dysentery. Slime molds behave similarly to an
ameba, but they aggregate to form much larger complex structures MicroByte
during part of their life cycle. Heteroloboseans exist in an ameboid A single paramecium (a protozoan) can ingest as many as 5 million
form during part of their life cycle, but also form flagellated cells. bacteria in one day.
Naegleria fowleri can swim through the water in its flagellated
form but assumes its ameboid form upon entering the human
body, where it literally eats the brain of its host (figure 12.14). Protozoan Reproduction
Entamoeba histolytica, p. 605 Naegleria fowleri, p. 663 Some protozoa have complex life cycles involving more than
one habitat or host. A single protozoan species that can exist
as a trophozoite (vegetative or feeding form) or cyst (rest-
Structure of Protozoa ing form) is polymorphic (figure 12.14). Environmental
conditions—such as the lack of nutrients, moisture, O2, low
By definition, protozoa are not photosynthetic and thus lack
temperature, or the presence of toxic chemicals—may trigger
chloroplasts. Protozoa also lack the rigid cellulose cell wall
the development of a protective cyst. Some protozoans, such
found in algae or the chitinous cell wall found in fungi.
as Cryptosporidium and Entamoeba, develop a protective cell
Foraminifera, however, secrete a hard shell of calcium carbon-
wall during the cyst stage of their life cycle. This protects
ate. Over millions of years these shells have formed signifi-
the organism as it moves from host to host and also helps it
cant limestone deposits, such as the White Cliffs of Dover on
withstand stomach acid as it enters a new host. Stomach acid
the coast of England. Even though protozoa typically lack a
may assist in removal of the cell wall so that the trophozoite
cell wall, they often have a specific shape determined by the
can emerge in time to infect the host’s intestines. Many para-
material beneath the plasma membrane.
sitic protozoans are transmitted to new hosts during their cyst
Traditionally, protozoa have been grouped by their mode
stage. cryptosporidiosis, p. 603 amebiasis, p. 605
of locomotion. Many have specialized structures for movement
Both asexual and sexual reproduction are common in pro-
such as cilia, flagella, or pseudopodia. As described in chapter 3,
tozoa and may alternate during the life cycle of some organ-
eukaryotic flagella and cilia are distinctly different from prokary-
isms. Binary fission takes place in many groups of protozoa.
otic flagella (see figures 3.37 and 3.50). flagella, p. 74
Some protozoa divide by multiple fissions, or schizogony, in
which the nucleus divides a number of times and then the cell
produces many single-celled organisms. Multiple fission of
Protozoan Habitats the asexual forms of Plasmodium in humans results in large
The majority of protozoa are free-living aquatic organisms. They numbers of parasites released into the host’s circulation at
are essential as decomposers in many ecosystems. Some species, regular intervals, producing the cyclic symptoms of malaria.
however, are parasitic, living on or in other host organisms. In malaria, p. 686

(a) (b) (c)

FIGURE 12.14 Polymorphism in a Protozoan Species of Naegleria may infect humans. (a) In human tissues, the organism exists in the form
of an ameba (10–11 μm at its widest diameter). (b) After a few minutes in water, the flagellate form appears. (c) Under adverse conditions, a cyst
is formed.
? What is the advantage of flagella in water?
294 Chapter 12 The Eukaryotic Members of the Microbial World

The slime molds and water molds are protists that were
TABLE 12.4 Protozoa of Medical Importance
once considered types of fungi. On the surface they may
Genus of look like fungi and they may act like fungi, but at a cel-
Disease- Disease Page for lular and molecular level, they are completely unrelated
rRNA Causing Caused by Additional to them (see figure 12.13). Fungi and water molds, in
Classification Protozoan Protozoan Information particular, are good examples of convergent evolution,
Apicomplexan Plasmodium Malaria p. 686 which occurs when two organisms develop similar char-
Toxoplasma Toxoplasmosis p. 709 acteristics independently because of adaptations to similar
environments.
Cryptosporidium Cryptosporidiosis p. 603
Cyclospora Cyclosporiasis p. 604
Diplomonad Giardia Giardiasis p. 602
Slime Molds
Parabasalian Trichomonas Trichomoniasis p. 635
Slime molds are terrestrial organisms composed of ameboid
Kinetoplastid Trypanosoma African sleeping p. 662
sickness
cells that live on soil, leaf litter, or the surfaces of decaying
vegetation, where they ingest organic matter by phagocytosis.
Heterolobosean Naegleria Primary amebic p. 663
They are important links in the terrestrial food chain because
meningoencephalitis
they ingest microorganisms and, in turn, serve as food for
Lobosean Entamoeba Amebiasis pp. 605,
larger predators.
(diarrhea) 663
There are two types of slime molds—cellular and
plasmodial:
■ Cellular slime molds have a vegetative form composed of
single, ameba-like cells. When they run out of food, the single
Medical Importance of Protozoa
cells aggregate into a mass of cells called a slug. Some single
The majority of protozoa do not cause disease, but those that cells then form a fruiting body, while others differentiate
do have significant impact on world health. Malaria has been into spores. These look very much like fungal fruiting bodies
one of the greatest killers of humans through the ages. Up to and spores (figure 12.15a). The model eukaryotic organism,
300 million people in the world still contract malaria each Dictyostelium discoideum is a cellular slime mold important
year, and 1 million die of it, most of them in Africa. Amebiasis for the study of aggregation and multicellular development.
(amebic dysentery) affects nearly 50 million people worldwide
■ Plasmodial slime molds are large multinucleated “super-
each year, claiming up to 100,000 victims. Cryptosporidium
amebas” that may easily reach 0.5 m in diameter. They
and Giardia are also among the leading causes of diarrhea
are widespread and readily visible in their natural environ-
worldwide. Trypanosomes that cause sleeping sickness have
ment due to their large size and often their bright color
made some regions of Africa uninhabitable for centuries.
(figure 12.15b). Following germination of haploid spores,
Table 12.4 lists some disease-causing protozoans and directs
the cells fuse to form a diploid cell in which the nucleus
you to where some of the most significant are discussed in
divides repeatedly, forming a multinucleated stage called a
later chapters.
plasmodium. The plasmodium oozes over the surface of
decaying wood and leaves, ingesting organic debris and
MicroAssessment 12.3 microorganisms. When food or water is in short supply, the
Protozoa are microscopic, single-celled, non-photosynthetic, motile plasmodium is stimulated to form spore-bearing fruiting bod-
organisms. They occupy a variety of habitats and are a very important ies, and the process begins again.
part of food chains.
7. What are some important diseases caused by protozoa? MicroByte
8. What is the role of a cyst in protozoan reproduction? Japanese researchers have shown that a slime mold takes the shortest
path between two nutrients.
9. Why might a parasitic protozoan lack mitochondria? +

Water Molds
12.4 ■ Slime Molds The water molds, or oomycetes, were once considered fungi.
and Water Molds They form masses of white threads on decaying material
(figure 12.16). Like fungi, they secrete digestive enzymes onto a
Learning Outcomes substrate and absorb small molecules for nutrients. The cytoplasm
8. Compare and contrast the two types of slime molds with one
in their filaments is continuous with many nuclei. However, water
another and with water molds. molds have cellulose in their cell walls rather than chitin. They
9. Explain how fungi and water molds provide an example of
lack chloroplasts and have flagellated reproductive cells.
convergent evolution. Oomycetes cause some serious diseases of food crops such
as late blight of potato and downy mildew of grapes. The late
 Part II The Microbial World 295

Spores FIGURE 12.15 Slime Molds (a) Fruiting body

of a cellular slime mold. (b) Life cycle of a plasmo-
dial slime mold.
Fruiting bodies
release spores
Germination ? What are the activities of a plasmodial slime
mold when it is in the plasmodial form?

Diploid cell

(a)

Fruiting body

(b) Plasmodium

blight of potato was the cause of the potato famine in Ireland in 12.5 ■ Multicellular Parasites:
the 1840s that sent waves of immigrants to the United States (see
A Glimpse of History). Helminths
Learning Outcomes
MicroAssessment 12.4
10. Explain how disease-causing helminths can be transmitted to
Cellular slime molds may exist as single cells or may form an humans.
aggregation called a slug for reproduction. Plasmodial slime molds
11. Describe a disease caused by a roundworm, a tapeworm, and a
exist as a multinucleated “super-ameba.” Both engulf food by
fluke.
phagocytosis and form fruiting bodies similar to those of fungi. Water
molds secrete enzymes and absorb organic nutrients, and they form
masses of white threads on organic matter. The helminths, which include nematodes (roundworms), cestodes
10. How are slime molds and water molds similar to, and different (tapeworms), and trematodes (flukes), cause disease by invading
from, fungi? the host’s tissues or robbing it of nutrients. These multicellular
11. What environmental conditions led to the convergent evolution parasites have been largely controlled in industrialized nations, but
of fungi and water molds? + they still cause suffering and death of many millions each year in
developing parts of the world.
Helminths enter the body in a number of ways. Hookworm lar-
vae, for example, live in the soil and can burrow through human skin.
They multiply in the human digestive tract and are eliminated with
feces. When sanitation is poor and people are bare-footed, the parasite
is easily transmitted. Hookworms can be found in about 740 million
individuals, mostly in tropical and subtropical regions.
Some helminths are eaten with food, as when cysts of the
nematode Trichinella spiralis are ingested in the flesh of animals.
Eating undercooked pork is the most common cause of trichinel-
losis. More often, helminth eggs are ingested on the surface of
foods. Children with pinworms (Enterobius vermicularis), for
example, may pick up eggs by touching their anus and transmit
them to a surface that can be touched by another person who then
handles food without proper handwashing.
Some helminths are transmitted through insect bites. One
example is Wuchereria bancrofti, a type of nematode that causes
FIGURE 12.16 Water Mold The white "threads" secrete digestive elephantiasis and is transmitted by mosquitoes. These worms
enzymes used to break down the organic compounds in this insect. lodge in the lymphatic vessels where they block lymphatic drain-
? How is a water mold like a fungus; how is it different? age. The buildup of fluid can cause massive swelling in various
296 Chapter 12 The Eukaryotic Members of the Microbial World

PERSPECTIVE 12.1
What Causes River Blindness?
Female black flies, sometimes called gnats to a human host, leading to river blindness, in the host that can damage sensitive tissues
or buffalo gnats, swarm around their hosts also called onchocerciasis. Once in a human and result in vision impairment or blindness.
and bite repeatedly to obtain blood needed host, the larvae reside in nodules and mature Ultimately, it is the bacteria that cause river
for development of their eggs. They require to adulthood in about a year, at which time blindness.
rapidly flowing water for development of their adult females produce millions of microfi- Efforts to control river blindness have
larvae and so are most often found near rivers. lariae. These can migrate through the skin focused on eliminating the black fly vector and
These flies are associated with a disease called where they can be picked up by another bite providing a medication called ivermectin that
river blindness that affects at least 18 million of a black fly to continue the life cycle. When targets the worm. Ivermectin reduces the num-
individuals, 99% of whom live in Africa. Of infestations are heavy, the microfilariae can ber of microfilariae for a few months, but does
those, the World Health Organization esti- also be found in the blood and in the eye. So not destroy the adults, so repeated medication
mates that about 270,000 are blind as a result are the Onchocerca larvae the real cause of is needed. A newer more effective approach is
of their infection and another 500,000 have the disease? to target the Wolbachia bacteria with common
impaired vision. River blindness is the second As microfilariae move throughout the antimicrobial medications such as doxycycline
leading cause of infectious blindness. (The bodies of their hosts, they carry with them a or tetracycline. This not only reduces effects of
leading cause is trachoma caused by the bac- bacterial population of Wolbachia pipientis the disease, but it sterilizes the worms so that
terium Chlamydia trachomatis.) But are the that is necessary for fertility and viability of they can no longer produce microfilariae, thus
black flies the cause of this disease? the worms. When the microfilariae reach the disrupting the life cycle.
When biting, the flies may transmit larvae eye and then die, they release the Wolbachia.
of a filarial nematode, Onchocerca volvulus, The bacteria cause an inflammatory response

parts of the body (figure 12.17). Another example is Onchocerca in humans, which are its definitive host. Humans may become an
volvulus—a nematode that is spread by flies and can cause river accidental or dead-end host if infected by a parasite that normally
blindness (see Perspective 12.1). completes its life cycle in another host. For example, swimmers
Some helminths have complex life cycles involving one or are sometimes infected with the larvae of flukes that typically
more intermediate hosts that house a sexually immature stage of complete their life cycles in fish or water birds. These flukes can-
the parasite and are necessary for its development. Snails serve as not complete their life cycle in humans, but when they burrow
an intermediate host for the fluke Schistosoma mansoni, the cause under the skin, they cause local inflammation called “swimmer’s
of schistosomiasis. Sexual reproduction of the parasite takes place itch.” Table 12.5 lists the major diseases caused by helminths.

MicroByte
Over 1 billion people worldwide are believed to carry the roundworm
Ascaris lumbricoides.

Roundworms
A roundworm, or nematode, has a cylindrical, tapered body with a
digestive tract that extends from the mouth to the anus. The nema-
tode Caenorhabditis elegans is a model eukaryotic organism that
has been the subject of numerous studies in genetics and develop-
ment because it matures quickly, its genome has been sequenced,
and all 959 body cells can be identified. Many nematodes are free-
living in soil and water. Others are parasites and produce serious
disease. Almost 30,000 nematode species have been identified,
but some estimate that there may be over 1 million species.
Ascariasis, caused by Ascaris lumbricoides, is the most com-
mon human disease caused by roundworms. Females may reach
45 cm long and produce more than 200,000 eggs per day that
are eliminated in the feces. Ingested eggs hatch in the digestive
tract, releasing immature worms that burrow into the bloodstream
(figure 12.18). When they reach the lungs, they can be coughed up
FIGURE 12.17 Elephantiasis Buildup of lymphatic fluid has and swallowed. When the immature worms again reach the intes-
occurred because worms block the vessels leading from the limbs. tine, they grow and begin to produce eggs that can again be released
? What type of worm causes this condition? with the feces. Although they do not feed on human tissue, they do
 Part II The Microbial World 297

TABLE 12.5 Nematodes, Cestodes, and Trematodes

Infectious Agents Disease Disease Characteristics

Nematodes (roundworms)
Pinworms (Enterobius vermicularis) Enterobiasis Anal itching, restlessness, irritability, nervousness, poor sleep
Whipworm (Trichuris trichiura) Trichuriasis Abdominal pain, bloody stools, weight loss
Hookworm (Necator americanus and Hookworm Anemia, weakness, fatigue, physical and intellectual disability in
Ancylostoma duodenale) disease children
Threadworm (Strongyloides stercoralis) Strongyloidiasis Skin rash at site of penetration, cough, abdominal pains, weight loss
Ascaria (Ascaris lumbricoides) Ascariasis Abdominal pain, live worms vomited or passed in stools
Trichinella (Trichinella spiralis) Trichinellosis Fever, swelling of upper eyelids, muscle soreness
Filaria (Wuchereria bancrofti and Brugia malayi) Filariasis Fever, swelling of lymph glands, genitals, and extremities
Cestodes (tapeworms)
Fish tapeworm (Diphyllobothrium latum) Tapeworm Few or no symptoms, sometimes anemia
disease
Beef tapeworm (Taenia saginata) Tapeworm Few or no symptoms, sometimes anemia
disease
Pork tapeworm (Taenia solium) Cysticercosis Variable symptoms depending on location and number of eggs that
form larval cysts (cysticerci) in the body
Trematodes (flukes)
Cercaria (Schistosoma mansoni) Schistosomiasis Liver damage, malnutrition, weakness, and accumulation of fluid in
the abdominal cavity
Cercaria of birds and other animals Swimmer’s itch Inflammation of the skin, itching

1 Worm eggs from contaminated

soil are ingested.

3 Larvae enter the lungs

from capillaries and can
then be coughed up
and swallowed.

Bronchiole
100 µm Alveolus

2 Ingested eggs hatch;

larvae penetrate
intestinal capillaries
and are carried to lungs.

4 In the intestine, mature larvae

develop into adult worms.

5 Eggs released from adult worms

are passed in the feces.

FIGURE 12.18 Ascariasis Life cycle of Ascaris lumbricoides, the largest roundworm infesting the human intestine, reaching 30–40 cm.
Larvae hatching in the intestine migrate through the lungs and back to the intestine before maturing to adulthood. The thick-walled
ova are nearly spherical.
? How does Ascaris get from the lungs to the intestines?
298 Chapter 12 The Eukaryotic Members of the Microbial World

FIGURE 12.19 Scolex attached

Pork Tapeworm The to intestinal wall
scolex holds the tape-
Scolex Hooklets
worm to the intestinal
surface. Proglottids
contain reproductive
structures. They are Sucker
shed in the feces as the Repeated
proglottid
tapeworm elongates by
sections
adding new segments.
? Why doesn’t a
tapeworm need a 200 µm
digestive system?

rob the body of nutrients by feeding on material that passes MicroAssessment 12.5
through the digestive tract. They may cause choking and pulmo-
nary symptoms when they enter the respiratory tract. Helminths, including the roundworms, tapeworms, and flukes, cause
serious diseases in humans. They may enter a human host through
ingestion with food or water, by an insect bite, or by piercing the skin.
Tapeworms Many helminths have a complex life cycle with more than one host.
12. What are the major differences among nematodes, cestodes, and
Tapeworms, or cestodes, have flat, ribbon-shaped bodies and some
trematodes?
types can reach over a meter in length. They have no digestive
13. Differentiate between a definitive host and an intermediate host.
system, and do not feed directly on the tissues of their host. Rather,
adult tapeworms attach within the intestines of the definitive host 14. Why do so many helminth diseases occur in the tropics? +
and absorb predigested nutrients through their body. The head end
(scolex) often has suckers and hooks for attachment (figure 12.19).
Behind the scolex are a number of segments called proglottids
that contain both male and female reproductive structures. The 12.6 ■ Arthropods
segments farthest from the scolex contain fertilized eggs. As
the worm grows, these segments break off and are eliminated in Learning Outcomes
the feces along with tapeworm eggs. When a suitable intermediate 12. Explain how arthropods are related to disease in humans.
host ingests the eggs, the eggs hatch, releasing a larval form that 13. Give an example of a disease related to flies, mosquitoes,
penetrates the intestinal wall and migrates into tissues. These larval fleas, lice, ticks, and mites.
forms are infectious when consumed by a definitive host.
The most common tapeworms of humans have intermediate Arthropods include the insects (such as flies, mosquitoes, lice, and
hosts of cattle, pigs, and fish. Humans (the definitive host) become fleas) and the arachnids (such as ticks and mites). Their main role
infected when they eat raw or undercooked meat containing the in disease is to serve as vectors that can transmit microorganisms
larval forms. Unfortunately, humans can also serve as the acci- and viruses to humans. An arthropod may act as a mechanical vec-
dental intermediate host of the pork tapeworm (Taenia solium). If tor that simply transfers a pathogen from one surface to another, or
someone inadvertently ingests eggs of this tapeworm, the eggs can it may be an essential part of the life cycle of the pathogen, acting
hatch, releasing larvae. These may migrate to the brain resulting in as a biological vector. For example, species of Plasmodium that
serious neurological symptoms. cause malaria multiply within an Anopheles mosquito during their
life cycle, and species of trypanosomes that cause African sleeping
sickness multiply within the tsetse fly (Glossina sp.). vector, p. 442
Flukes malaria, p. 686 African sleeping sickness, p. 662
Flukes, or trematodes, are flat leaf-shaped organisms with Animals such as mammals or birds may be hosts in the life
suckers that hold them in place while sucking fluids from the host. cycle of a pathogen and act as reservoirs for it. When an arthropod
Schistosoma mansoni, a blood fluke, is the most common cause vector feeds on a reservoir host, it may pick up pathogens and then
of schistosomiasis worldwide, resulting in about 20,000 deaths per transfer them to humans in a later bite. Some arthropods bite only
year. Female flukes are held by the larger male and lay eggs con- one type of host. Certain mosquitoes that carry Plasmodium typi-
tinually in blood vessels near the intestine. Inflammatory reactions cally bite only humans. However, fleas that carry Yersinia pestis,
cause the blood vessels to rupture into the intestine, releasing eggs the bacterium responsible for plague, bite both humans and small
that are carried from the body in feces. Larvae develop in fresh mammals such as rats. reservoir, p. 438
water and are taken up by a certain species of snail. After asexual The incidence of vector-borne diseases can be decreased by
reproduction, tail-bearing larvae are released from the snail and controlling the vector or a reservoir host. The risk of mosquito-
can penetrate the skin of a human host wading in the water. The borne encephalitis, for example, can be minimized by eliminating
larvae enter blood vessels, reproduce sexually in the liver, and standing water and using insect repellant. These do not act on the
begin the cycle again. Schistosomiasis infections do not occur in virus that causes encephalitis, but they do reduce the incidence
the United States as there is no appropriate snail intermediate host. of vector transmission. Plague in the United States has been
 Part II The Microbial World 299

TABLE 12.6 Some Arthropods That Transmit Infectious Agents

Infectious Page for More
Arthropod Agent Disease and Characteristic Features Information

Insects
Tsetse fly (Glossina species) Trypanosomes African sleeping sickness—sleepiness, headache, coma p. 662
Sand fly Leishmania Leishmaniasis—ulcers, nosebleeds, diarrhea, fever, cough p. 521
(Phlebotomus species)
Black fly (Simuliidae species) Onchocercus Onchocerciasis—rash, itching, visual impairment p. 296
Mosquito (Anopheles species) Plasmodium species Malaria—chills, bouts of recurring fever p. 686
Mosquito (Culex species) Togavirus Equine encephalitis—fever, nausea, convulsions, coma p. 655
Mosquito (Aedes aegypti) Flavivirus Yellow fever—fever, vomiting, jaundice, bleeding p. 682
Mosquito (Aedes aegypti) Flavivirus Dengue fever—high fever; headache; joint, muscle, and p. 683
bone pain
Flea (Xenopsylla cheopis) Yersinia pestis Plague—fever, headache, confusion, enlarged lymph nodes, p. 678
skin hemorrhage
Louse (Pediculus humanus) Rickettsia Typhus—fever, hemorrhage, rash, confusion p. 521
prowazekii
Arachnids
Tick (Dermacentor species) Rickettsia rickettsii Rocky Mountain spotted fever—fever, hemorrhagic rash, p. 529
confusion
Tick (Ixodes species) Borrelia burgdorferi Lyme disease—fever, rash, joint pain, nervous system impairment p. 531

controlled mostly by eliminating rat populations that may infect hollow feeding tube through the host’s skin to the subcutaneous
their fleas with the bacterium Yersinia pestis. Cases of plague capillaries (figure 12.20). She pumps saliva through the tube,
today occur from occasional transmission by fleas on wild rodents which increases blood flow and prevents clotting of the victim’s
such as rock squirrels or prairie dogs. Examples of some important blood. She can take in as much as twice her body weight in blood,
arthropods, the agents they transmit, and the resulting diseases are also picking up infectious agents circulating within the host’s
shown in table 12.6. plague, p. 678 capillaries. These agents multiply within the mosquito’s body and
can later be transferred to a new host in a subsequent bite. yellow
Mosquitoes fever, p. 682 dengue fever, p. 683

Mosquitoes are insects known to transmit diseases such as

malaria, yellow fever, dengue fever, and West Nile encephalitis. MicroByte
The itch of a mosquito bite is caused by an allergic reaction to the
A female mosquito needs a blood meal for the proper develop- mosquito’s saliva.
ment of her eggs. During a bite, the mosquito forces a sharp,

Storage area Midgut Cysts of

malaria parasite
Antenna

Stomach

Mouthparts Salivary Storage Body

glands area cavity

FIGURE 12.20 Internal Anatomy of a Mosquito Storage areas allow ingestion of large amounts of blood, and salivary glands discharge
pathogens into the host. The Anopheles mosquito adopts a curious head-stand posture when feeding.
? Is the mosquito an intermediate host or the definitive host for the organism that causes malaria?
300 Chapter 12 The Eukaryotic Members of the Microbial World

The pubic louse, Phthirus pubis, is commonly transmitted

during sexual intercourse. It is not a vector of infectious disease,
but it can cause an unpleasant itch associated with “crabs.”

Ticks
Ticks are arachnids. Arachnids can be distinguished from insects
because they lack wings and antennae, their thorax and abdomen
are fused, and adults have four pairs of legs rather than three.
Ticks generally live in low vegetation where they may contact a
suitable host passing by. Once in contact with a host, the tick bur-
rows into skin with its mouthparts (see figure 22.9). When a tick
attaches in the scalp or is very small, it may go unnoticed for days,
FIGURE 12.21 Pediculus humanus A body louse, which is the during which time it continually feeds from its host.
vector for several human diseases. Dermacentor andersoni, the wood tick, is the vector for
? Can body lice act as mechanical vectors?
Rocky Mountain spotted fever caused by the bacterium Rickettsia
rickettsii. Another tick, Ixodes scapularis, transmits with its saliva
Borrelia burgdorferi, the spirochete that causes Lyme disease. A
neurotoxin in the saliva of some ticks can cause muscle weakness,
a condition called tick paralysis. This occurs especially in chil-
Fleas dren but recovery is rapid following removal of the tick. Rocky
Mountain spotted fever, p. 529 Lyme disease, p. 531
Fleas are wingless insects, but their appendages are adapted for
easily jumping up to 30 cm at a time. They are generally more of
a nuisance than a health hazard, but they can transmit a number of Mites
pathogens, including the bacterium Yersinia pestis, which causes Mites, like ticks, are arachnids. Microscopic mites of the genus
plague. Fleas pick up the pathogen when biting an infected host, Demodex typically live in the hair follicles or oil-producing
and the bacteria multiply within the digestive tract of the flea, glands without being noticed. Large numbers of dust mites often
causing a blockage. The starving flea bites repeatedly, each time live indoors and feed on organic material such as shed skin cells.
passing bacteria from its blocked digestive tract to a host. Fleas Although they do not transmit infectious disease, inhalation of the
can live in vacant buildings in a dormant stage for many months. mites and their waste products can sometimes trigger asthma. The
When the building becomes inhabited, the fleas quickly mature larvae of some mites are called “chiggers” and may cause intense
and hungrily jump to greet the new hosts. itching where they attach and feed on fluids within skin cells.
Scabies, a disease caused by a mite, Sarcoptes scabiei,
Lice is characterized by an itchy rash most prominent between the
fingers, under the breasts, and in the genital area. Scabies
Like fleas, lice (singular: louse) are small, wingless insects that
prey on mammals and birds by piercing their skin and suck-
ing blood. The appendages of lice, however, are adapted for
attachment rather than jumping. Pediculus humanus, the most
notorious of the lice, is 1 to 4 mm long, with a membrane-like
lip housing tiny teeth that anchor it firmly to the skin of the host
(figure 12.21). Lice have a piercing apparatus similar to that of
fleas and mosquitoes used for obtaining a blood meal. Pediculus Skin surface
humanus has only one host—humans—but easily spreads from
one person to another by direct contact or by contact with personal
items, especially in areas of crowding and poor sanitation. An
infestation of lice is called pediculosis.
The two subspecies of Pediculus humanus are popularly
Epidermis
termed body lice and head lice. Body lice can transmit trench
fever, caused by the bacterium Bartonella quintana; epidemic
typhus, caused by the bacterium Rickettsia prowazekii; and relaps-
ing fever caused by the bacterium Borrelia recurrentis. Trench
fever occurs sporadically among severe alcoholics and the home-
less of large American and European cities. Head lice do not trans- FIGURE 12.22 Sarcoptes scabiei (Scabies Mite) The female
mit disease, but they are easily transmitted from human to human. burrows into outer skin layers to lay her eggs, causing an intensely
They are most often discovered when eggs resembling dandruff, itchy rash.
or nits, are found clinging to the scalp or hair. ? Is a mite an insect or an arachnid?
 Part II The Microbial World 301

is easily transmitted by personal contact, and the disease is MicroAssessment 12.6

commonly acquired during sexual intercourse. Female mites
burrow into the outer layers of epidermis (figure 12.22), feed- Arthropods such as flies, mosquitoes, fleas, lice, ticks, and mites act
as vectors for the spread of disease. Some participate in the life cycle
ing and laying eggs over a lifetime of about 1 month. Allergic of an infectious agent and transmit disease through saliva when biting
reactions are largely responsible for the itchy rash of scabies. or burrowing. Others infest the skin and cause itching.
Diagnosis requires locating the mites, since scabies mimics
15. How can arthropods spread disease in humans?
other skin diseases. Sarcoptes scabiei is not known to transmit
16. Name two diseases transmitted by ticks.
infectious agents. Mites of domestic animals and birds can
also cause an itchy rash in humans. 17. Why are diseases transmitted by insect vectors more common
in the summer than in the winter? +

FUTURE CHALLENGES 12.1

The Continued Fight to Eradicate Malaria
Malaria has affected humans for millennia. Gates Foundation, among others, has pledged foundations, and governments worldwide. As
The symptoms were described in ancient over a billion dollars to aid in the effort. A the battle continues, mosquitoes will become
Chinese writings more than 4,500 years ago. multifaceted approach includes distribution of resistant to pesticides and Plasmodium will
Each year about a million people still die insecticide-treated bed nets, spraying interior become increasingly resistant to antimalarial
from malaria; most are in Africa and most are buildings with insecticide, eliminating breed- drugs. Researchers will have to work hard to
children under the age of 5 years. With drug- ing grounds for mosquitoes, and providing stay ahead of both. To truly eradicate malaria,
resistant strains spreading rapidly, more peo- effective and affordable drugs. These efforts a vaccine is needed. Clinical trials are under-
ple die from malaria today than 30 years ago. have met with some success. For example, the way for a vaccine that could reduce malaria
The economic costs to the people living in incidence of malaria decreased by 50% in 26 deaths by 50%, but it is not expected to be
malaria-infested parts of the world are huge. countries between 2000 and 2006. generally available until 2015. Newer vac-
Drug companies and foundations are continu- In 2010, 39 countries were working to cines will attempt to disrupt the life cycle of
ing their effort to develop new vaccines and eradicate malaria. This will require billions the parasite by preventing reproduction in the
drugs to treat malaria. The Bill and Melinda of dollars along with support of corporations, mosquito vector.

Summary
THE EUKARYOTIC MEMBERS a wide range of temperatures, and at pH from 2.2 to 9.6. Fungi can
OF THE MICROBIAL WORLD degrade most organic materials.

Algae, fungi, and protozoa are not accurate classification terms when Symbiotic Relationships of Fungi
the rRNA sequences of these organisms are considered. Cell structure in Lichens result from an association of a fungus with a photosynthetic
eukaryotic organisms is different from that seen in prokaryotes. Eukaryotes organism such as an alga or a cyanobacterium (figure 12.5). Mycorrhizas
have a membrane-bound nucleus. Reproduction may be asexual using are the result of an intimate association of a fungus and the roots of
mitosis or sexual using meiosis, which forms gametes (figure 12.1). a plant (figure  12.6). Leaf-cutter ants grow gardens of fungus for food
(figure 12.7).
12.1 ■ Fungi Reproduction in Fungi
Yeast, mold, and mushroom are common terms that indicate morpho-
Asexual reproduction may occur by a variety of spore-producing
logical forms of fungi (figure 12.2). Fungi have chitinous cell walls and
structures, by budding, or by fragmentation (figures  12.8, 12.9). Sexual
are often saprophytes, secreting enzymes onto a surface and absorbing
reproduction may involve fusion of hyphae from different mating types.
nutrients.
Economic Importance of Fungi
Types of Fungi (table 12.1)
Classification of fungi is in flux. Zygomycetes (figure 12.3), ascomycetes, The yeast Saccharomyces is used in the production of beer, wine, and
basidiomycetes, and chytridiomycetes are distinctive types of fungi. bread. Penicillium and other fungi synthesize antibiotics. Fungi spoil
many food products and cause diseases of plants such as Dutch elm
Structure of Fungi disease and wheat rust. Fungi have been useful tools in genetic and
Fungal filaments are called hyphae, and a group of hyphae is called a biochemical studies.
mycelium (figure 12.4). Dimorphic fungi can grow either as a single cell
Medical Importance of Fungi (table 12.2)
(yeast) or as mycelia.
Relatively few fungi cause human disease, but they cause devastat-
Fungal Habitats ing diseases in plants. Fungi may produce an allergic reaction. Fungi
Fungi inhabit just about every ecological habitat and can spoil a large cause mycoses such as candidiasis. Fungi can produce toxins that make
variety of food materials because they can grow in high concentrations humans ill. These include ergot, those in poisonous mushrooms, and
of sugar, salt, and acid. Fungi can be found in moist environments, at aflatoxin.
302 Chapter 12 The Eukaryotic Members of the Microbial World

12.2 ■ Algae 12.4 ■ Slime Molds and Water Molds

Algae are a diverse group of photosynthetic organisms that contain Slime molds and water molds were once considered types of fungi.
chlorophyll a.
Slime Molds (figure 12.15)
Types of Algae (table 12.3) Cellular slime molds exist as ameba-like single cells, but when
Types of algae differ in their major photosynthetic pigments. Organisms food supplies run low, they aggregate into a slug in which some
are placed on the phylogenetic tree according to rRNA sequences cells differentiate into spores. Plasmodial slime molds form a
(figure 12.10). multinucleated plasmodium that oozes over a surface, ingesting
organic material. When food runs short, they form fruiting bodies
Structure of Algae that bear spores.
Algae may be microscopic (figure  12.11) or macroscopic (figure  12.12).
Water Molds
Their cell walls are made of cellulose and other commercially important
materials such as agar and alginic acid. Oomycetes, also known as water molds, cause some serious diseases
of plants (figure  12.16). Water molds and fungi are an example of
Algal Habitats convergent evolution.
Algae are found in fresh and salt water as well as in soil. Unicellular
algae make up a significant part of the phytoplankton. 12.5 ■ Multicellular Parasites: Helminths (table 12.5)
Helminths can be transmitted by burrowing through the skin, being
Algal Reproduction ingested, or being transmitted through insect bites (figure 12.17). Some
Algae reproduce asexually as well as sexually. helminths have complex life cycles with asexual stages occurring
in intermediate hosts and the sexual or adult stage occurring in the
Medical Importance of Algae
definitive host. Humans may be a dead-end host in which the organ-
Algae do not directly cause disease, but produce toxins during algal ism cannot complete its life cycle.
blooms that are ingested by fish and shellfish. When these fish and
shellfish are eaten by humans, they may develop paralytic shellfish Roundworms
poisoning with dizziness, muscle weakness, and even death may result; Most nematodes or roundworms are free-living, but they may cause
cooking does not destroy the toxins. serious disease such as pinworm disease, hookworm disease, and asca-
riasis (figure 12.18).
12.3 ■ Protozoa Tapeworms
Protozoa are a diverse group of microscopic, unicellular organisms that Cestodes are tapeworms with segmented bodies and hooks to attach to
lack chlorophyll. the wall of the intestine (figure 12.19). Most tapeworm infections occur
in persons who eat uncooked or undercooked meats.
Types of Protozoa (figure 12.13)
In classification schemes based on rRNA, protozoa are not a sin- Flukes
gle group of organisms. Apicomplexans (Plasmodium, Toxoplasma, Trematodes, or flukes, often have complicated life cycles that neces-
Cryptosporidium, Cyclospora) are parasites with an apical complex sarily involve more than one host. Schistosoma mansoni larvae can
that helps them to penetrate host cells. Diplomonads (Giardia) and penetrate the skin of persons wading in infected waters and cause seri-
parabasalids (Trichomonas) have no mitochondria. Kinetoplastids ous disease.
(Trypanosoma, Leishmania) have distinct mitochondrial DNA.
Loboseans (Entamoeba) and heteroloboseans (Naegleria) move by 12.6 ■ Arthropods
pseudopodia at some stage in their lives (figure 12.14). Arthropods act as vectors for disease (table 12.6).
Structure of Protozoa Mosquitoes
Protozoa lack a cell wall, but most maintain a definite shape using the Mosquitoes spread disease by picking up disease-causing organisms
material lying just beneath the plasma membrane. when the mosquito bites and later injecting these organisms into subse-
quent animals that it bites (figure 12.20).
Protozoan Habitats
Fleas
Most protozoa are free-living and are found in marine and fresh water
as well as terrestrial environments. They are important decomposers in Fleas transmit disease such as plague.
many ecosystems and are a key part of the food chain. Lice
Protozoan Reproduction Lice can transmit trench fever, epidemic typhus, and relapsing fever
(figure 12.21).
Life cycles are often complex and include more than one habitat or host.
Some are polymorphic with a vegetative trophozoite form and a rest- Ticks
ing cyst form. Reproduction is often by binary fission; some reproduce Ticks are implicated in Rocky Mountain spotted fever and Lyme
by multiple fissions or schizogony. disease.
Medical Importance of Protozoa (table 12.4) Mites
Protozoa cause diseases such as malaria, African sleeping sickness, Mites cause scabies, and dust mites are responsible for allergies and
toxoplasmosis, and amebic dysentery. asthma (figure 12.22).
 Part II The Microbial World 303

Review Questions
Short Answer 7. Body lice
1. What are the major differences between a prokaryotic cell and a a) can act as a vector to transmit disease.
eukaryotic cell? b) are not infectious.
2. What are the differences among a yeast, a mold, and a mushroom? c) have eight legs and sucking mouthparts.
3. How do mycorrhizas improve the growth of a green plant? d) are more closely related to ticks than they are to mosquitoes.
4. In what ways are fungi economically important? 8. All algae have
5. What is a mycosis? Give an example. a) chlorophyll a.
6. What characteristics do all algae have in common? b) cell walls that contain agar.
7. Compare and contrast the organisms that cause malaria and c) holdfasts.
African sleeping sickness and their transmission. d) red tides.
8. Name a disease for which humans are an intermediate host and 9. Which of the following statements regarding protists is false?
another for which humans are a definitive host. Give an example a) They include both autotrophic and heterotrophic organisms.
of a disease in which humans are a dead-end host. b) They include both microscopic and macroscopic organisms.
9. Describe the life cycle of Schistosoma mansoni. c) They often act as vectors in disease transmission.
10. Explain how a fly might act as a mechanical vector for one disease d) They include algae and protozoa.
and a biological vector for another. 10. Which of the following statements regarding tapeworms is false?
a) They absorb nutrients from the host through their body wall.
Multiple Choice
b) They complete their life cycles in a single host.
1. Members of this group have chitinous cell walls. c) They can form cysts in the tissue of their host.
a) Algae b) Protozoa c) Fungi d) They cannot be transmitted from human to human.
d) Helminths e) Arthropods
2. Members of this group are photosynthetic. Applications
a) Algae b) Protozoa c) Fungi 1. A molecular biologist working for a government-run fishery
d) Helminths e) Arthropods in Vietnam is interested in controlling Pfisteria in fish farms.
3. This group helps produce many of the foods that we eat. Pfisteria produces toxins that stun the fish and then causes the skin
to slough off, allowing the dinoflagellates to dine on the tissues of
a) Algae b) Protozoa c) Fungi
the fish. He needs to develop a treatment that kills Pfisteria without
d) Helminths e) Arthropods harming the fish or the beneficial green algae that serve as food
4. Protozoa reproduce asexually by for the young fish. What strategy should the biologist consider for
a) schizogony. developing a selective treatment?
b) fragmentation. 2. Paper recycling companies refuse to collect paper products that are
c) meiosis. contaminated with food or have been sitting wet for a day. A col-
d) polymorphism. lege sorority member who is running a recycling program on campus
5. Which of the following is mismatched? wishes to know the reason for this. What reason did the chemist who
works for the recycling company probably give her for this policy?
a) Plasmodium—malaria
b) Trypanosomes—dysentery
Critical Thinking +
c) Dinoflagellates—paralytic shellfish poisoning
1. If you discover a new type of nucleated cell in a lake near your
d) Nematode—trichinellosis
home, how would you determine whether the cell is from a fungus,
6. Which of the following is mismatched? an alga, a protozoan, or a water mold?
a) Trematode—fluke 2. Fungi are known for growing and reproducing in a wide range of
b) Tick—arachnid environmental extremes in temperature, pH, and osmotic pres-
c) Baker’s yeast—algae sure. What does this tolerance for extremes indicate about fungal
d) Apicomplexan—protozoa enzymes?
13 Viruses, Viroids, and Prions
KEY TERMS
Bacteriophage A virus that infects
bacteria; often shortened to phage.
Latent Infection Viral infection
in which the viral genome is present
Plaque Assay Method used to
measure the number of viral particles
present in a sample.
Prion An infectious agent that
but not active, so viral particles are causes a neurodegenerative disease;
not being produced. consists of protein similar in amino
acid sequence to a normal protein in
Lysogen A bacterium that carries
the body.
phage DNA (the prophage)
integrated into its genome. Productive Infection Viral
infection in which more viral
Lysogenic Conversion A change
particles are produced.
in the properties of a bacterium
conferred by a prophage. Viroid An infectious agent of plants
that consists only of RNA.
Lytic Infection Viral infection
of a host cell with a subsequent Virion A complete virus in its inert
production of more virus particles non-replicating form; also referred to
and lysis of the cell. as a viral particle.

replication, or motility. When a viral genome finds its way into

SV40 (simian virus 40), a virus of monkeys.
a host cell, however, it can hijack that cell’s replication machin-
ery, inducing the cell to produce more virus particles. In essence,
viruses straddle the definition of life. Outside of a cell they are
inert, but inside a cell they direct activities that have a profound
A Glimpse of History effect on the cell. Most scientists agree, however, that viruses do
During the late nineteenth century, many bacteria, fungi, and protozoa not fit the definition of life. So although they are infectious agents,
were identified as infectious agents. Most of these organisms could be they are not organisms.
easily seen with the aid of a microscope and grown in the laboratory. Viruses can be broadly grouped into two general types
In the 1890s, however, D. M. Iwanowsky and Martinus Beijerinck based on the category of cells they infect. Some infect pro-
found that mosaic disease—a disease of tobacco plants—was caused karyotic cells, and others infect eukaryotic cells. Although
by an unusual agent. The agent was too small to be seen with the light both groups are viruses, those that infect bacteria are also
microscope, passed through filters that retained most known bacteria, referred to as bacteriophages, or simply phages (phage means
and could be grown only in media that contained living cells. Beijerinck
“to eat”).
called the agent a filterable virus. About 10 years later, F. W. Twort in
England and F. d’Herelle in France discovered a “filterable virus” that
The fact that viruses are obligate intracellular parasites makes
destroys bacteria. Virus means “poison,” a term once applied to all infec- them very difficult to study. Unlike most bacteria and eukaryotic
tious agents. With time, the adjective filterable was dropped and only the cells, which can be grown in pure culture, viruses require live
word virus was retained. organisms as hosts. To add to this difficulty of studying viruses,
Viruses have many features more characteristic of complex chemi- they are too small to be seen with a light microscope and can be
cals than cells. For example, tobacco mosaic virus (TMV) can be pre- visualized only with an electron microscope.
cipitated from a suspension with ethyl alcohol and still remain infective. The first section of this chapter describes the general
A similar treatment destroys the infectivity of bacteria. Further, in 1935, characteristics of viruses. The primary focus of the remaining
Wendell Stanley of the University of California, Berkeley, crystallized sections is on bacteriophages and animal viruses. The latter are
TMV. Its physical and chemical properties obviously differed from those obviously important because of the diseases they cause, but the
of cells, which cannot be crystallized. Surprisingly, the crystallized TMV
reasons for learning about bacteriophages may not be so appar-
could still cause disease.
ent. We study them because they are easy to cultivate in the
laboratory, and they serve as an important model to understand

I
n the simplest of terms, viruses can be viewed as genetic the molecular biology and relationships of animal viruses with
information—either DNA or RNA—contained within a protec- their hosts. What you learn about them will help you understand
tive coat. They are inert particles, incapable of metabolism, similar relationships between the medically important viruses

304
 Part II The Microbial World 305

and the cells they infect. Bacteriophages are also important 13.1 ■ General Characteristics
because they serve as a vehicle for horizontal gene transfer in
bacteria, which was described in chapter 8. In addition, they of Viruses
are important because they kill bacteria, thereby limiting bac-
terial populations in nature. This is significant ecologically, Learning Outcomes
but also has medical applications. In 2006, the Food and Drug 1. Describe the general features of viral architecture.
Administration approved a preparation of bacteriophages that 2. Describe how viruses are classified and named.
infect the food-borne pathogen Listeria monocytogenes for
use as an antimicrobial agent on ready-to-eat meat and poultry Most viruses are notable for their small size (figure 13.1). They
products. L. monocytogenes multiplies at refrigeration tempera- are approximately 100- to 1,000-fold smaller than the cells they
tures and causes a potentially lethal disease, which is why it is a infect. The smallest viruses are about 10 nm in diameter, and
concern on products that are eaten without cooking. The use of contain very little nucleic acid, perhaps as few as 10 genes. The
bacteriophages as an alternative to antibiotics in treating bacte- largest known viruses are about 800 nm, the size of the smallest
rial infections has also been explored. horizontal gene transfer, bacterial cells. Indeed, one very large virus is so big it was first
p. 200 Listeria monocytogenes, p. 648 identified as a bacterium (see Perspective 13.1).

Tobacco mosaic virus

(250 nm)
Adenovirus
(90 nm)
Hepadnavirus
(42 nm)
Poliovirus
(30 nm)
T4 bacteriophage
(225 nm)

Mimivirus
(800 nm)

Human red blood cell

(10,000 nm diameter) E. coli
(3,000 × 1,000 nm)

FIGURE 13.1 Relative Sizes of Viruses

? Why are viruses so much smaller than the cells they infect?
306 Chapter 13 Viruses, Viroids, and Prions

PERSPECTIVE 13.1
Microbe Mimicker
Like bacteria, viruses vary tremen- the new virus Sputnik and called it
dously in size and complexity. This a virophage, and the infected virus
is becoming increasingly apparent a mamavirus (figure 1). Sputnik
as viruses from a variety of animal can replicate in the ameba only
and bacterial hosts are studied in when mimivirus has infected the
greater detail. Perhaps the most same cell. When this happens, fewer
unusual virus recently characterized mimivirus particles are produced
came from Acanthamoeba, a free- and they display unusual morphol-
living ameba that may cause certain ogies. Thus, Sputnik behaves as a
forms of pneumonia. The virus was true parasite, the first of its kind
first thought to be a Gram-positive but probably not the last. Sputnik
bacterium because of its staining 100 nm
appears to contain genes from
characteristics and hairy appear- other viruses, raising an intriguing
FIGURE 1 Mamavirus (Large Mimivirus) with Sputnik
ance, with many fibrils sticking out question—is horizontal gene trans-
Virophages
from its capsid (see figure 13.1). fer between viruses a possibility?
Its name, mimivirus, reflects this horizontal gene transfer, p. 200
original misconception (mimicked a microbe). repair, translation, and polysaccharide biosyn- The mimivirus and virophages are unlikely
Mimivirus is the largest DNA virus ever thesis. However, like all viruses, the mimi to be the last unusual viruses discovered. Many
characterized—the diameter of its capsid is virion does not undergo cell division nor does it potential host organisms have not yet been
approximately 800 nm. This is twice the size contain ribosomes. This virus has been placed in cultured, and thus many likely virus families
of a small bacterium such as Mycoplasma. a taxonomic group of other large DNA viruses, remain undiscovered. Their discovery and char-
Its genome size is enormous for a virus—1.2 members of which infect distinctly different acterization will not only expand our knowledge
million base pairs, which is large enough to organisms, including vertebrates and algae. of viruses but also will challenge our definition
encode almost 1,000 proteins. These genes Recently, a mimivirus-infected ameba was of what is living and non-living—and perhaps
encode proteins never before seen in viruses and shown to be infected with an additional virus, provide an answer to the question, “Where did
include enzymes of nucleic acid synthesis, DNA a parasite of mimivirus. Its discoverers named viruses originate?”

Viral Architecture
Capsomere
At a minimum, a virion (viral particle) consists of nucleic acid subunits
surrounded by a protein coat (figure 13.2). The protein coat,
Nucleic acid
called a capsid, protects the nucleic acid from enzymes and toxic
chemicals in the environment. It also carries any enzymes required Nucleocapsid
Capsid (entire
by the virus for infection of host cells. The capsid together with protein coat)
the nucleic acid it encloses is called the nucleocapsid. Given the
fact that viral genomes are relatively small in size, and therefore Spikes
can encode only a limited number of different proteins, it is not
surprising that capsids are simple in chemical structure. A cap- (a) Naked virus
sid is composed of identical protein subunits, called capsomers,
arranged in a precise manner to form the capsid. Spikes
Some viruses have an envelope, a lipid bilayer outside of the
capsid (figure 13.2b). How these enveloped viruses obtain the
envelope from the host cell will be described later. Sandwiched Matrix protein
between the nucleocapsid and envelope is the matrix protein,
which is unique to enveloped viruses. Viruses that do not have an Nucleic acid
envelope are called naked viruses. Nearly all phages are naked. Nucleocapsid Capsid (entire
In general, enveloped viruses are more susceptible to disinfectants protein coat)
because these chemicals damage the envelope, making the viruses
non-infectious. Envelope
Viruses contain only a single type of nucleic acid—either
RNA or DNA—but never both. This provides a useful method
(b) Enveloped virus
for classifying viruses, which are frequently referred to as either
RNA or DNA viruses. The genome may be linear or circular, FIGURE 13.2 Two Different Types of Viral Architecture
either double-stranded or single-stranded. The type of genome ? What components must all viruses have?
 Part II The Microbial World 307

has important implications with respect to the virus’s replication surface is actually 20 flat triangles arranged in a manner some-
strategy, and will be discussed in more detail later. what similar to a soccer ball. This arrangement is an efficient
Viruses have specific protein components that allow the design for any container that uses identical subunits and requires
virion to attach (adsorb) to specific receptor sites on host cells. the least energy to assemble. Helical viruses appear cylindrical
Phages, for example, have tail fibers that attach to host cells, and when viewed with the electron microscope. Their capsomers are
many animal viruses have protein structures called spikes that arranged in a helix, somewhat similar to a spiral staircase. Some
stick out from either the lipid bilayer of enveloped viruses or the helical viruses are short and rigid, whereas others are long and
capsid of naked viruses (see figures 13.1 and 13.2). filamentous. Complex viruses have more intricate structures.
A virus generally is one of three different shapes—icosahedral, Phages are the most common examples of this, many having an
helical, or complex (figure 13.3). Icosahedral viruses appear icosahedral nucleocapsid, referred to as the head, with a long heli-
spherical when viewed with the electron microscope, but their cal protein component, the tail.

FIGURE 13.3 Common

Icosahedral Shapes of Viruses
? What determines the shape
of the virus?

Protein coat
(capsid)
Nucleic acid

Adenovirus
75 nm
(a)

Helical

Nucleic acid

Protein coat
(capsid)

Tobacco mosaic virus (TMV) 100 nm

(b)

Complex

Protein coat
(capsid)
Nucleocapsid
Head with
nucleic acid
(DNA)
Tail

Base plate
Tail spike

Tail fibers T4 Bacteriophage 100 nm

(c)
308 Chapter 13 Viruses, Viroids, and Prions

Viral Taxonomy the key features, classification, and nomenclature of recognized

viruses. The ICVT 2009 report describes over 6,000 viruses
Although viruses are not living organisms, they are biological belonging to 2,288 species, 348 genera, 87 families, and 6 orders.
entities that are classified to provide easy identification and study Key characteristics used in the current classification scheme are
(table 13.1). The International Committee on Viral Taxonomy the genome structure (type of nucleic acid and strandedness) and
(ICVT) keeps an online database and publishes a report describing the hosts they infect (bacteria, archaea, animals, plants). A variety

TABLE 13.1 Classification of Some Human Viruses

Drawing of Virion Representative Member
Nucleic Acid Outer Covering Family (not to scale) (and disease caused)
DNA Viruses
Double- Naked Papillomaviridae Human papillomaviruses (some
stranded types cause warts; others cause
DNA cancers)

Polyomaviridae Merkel cell polyomavirus

(Merkel skin cancer)

Adenoviridae Human adenoviruses

(respiratory infections)

Enveloped Herpesviridae Herpes zoster virus

(chickenpox); herpes
simplex viruses
(cold sores, genital herpes)

Poxviridae Smallpox virus (smallpox)

Single- Naked Parvoviridae Human parvovirus B19

stranded (fifth disease)
DNA

RNA Viruses
Double- Naked Reoviridae Human rotaviruses
stranded (diarrheal disease)

Single- Naked Picornaviridae Polioviruses (poliomyelitis);

stranded rhinovirus (colds);
(plus strand) hepatitis A virus (hepatitis A)

Caliciviridae Norovirus (gastroenteritis)

Enveloped Togaviridae Rubella virus (rubella);

Chikungunya virus
(Chikungunya fever)

(continued)
 Part II The Microbial World 309

TABLE 13.1 Classification of Some Human Viruses (Continued)

Drawing of Virion Representative Member
Nucleic Acid Outer Covering Family (not to scale) (and disease caused)
RNA Viruses, cont.
Enveloped Flaviviridae Yellow fever virus (yellow
fever); dengue virus (dengue
hemorrhagic fever); hepatitis C
virus (hepatitis C)

Coronaviridae Coronavirus (SARS)

Single- Enveloped Rhabdoviridae Rabies virus (rabies)

stranded
(minus strand)

Filoviridae Ebola virus (hemorrhagic fever)

Paramyxoviridae Mumps virus (mumps); measles

virus (measles)

Orthomyxoviridae Influenza virus (influenza)

Bunyaviridae Hantavirus (hantavirus

pulmonary syndrome)

Arenaviridae Lassa virus (lassa fever)

Reverse Transcribing Viruses

DNA Enveloped Hepadnaviridae Hepatitis B virus (hepatitis B)

RNA Enveloped Retroviridae Human immunodeficiency virus

(AIDS)
310 Chapter 13 Viruses, Viroids, and Prions

TABLE 13.2 Grouping of Human Viruses Based on Route of Transmission

Virus Group Mechanism of Transmission Common Viruses Transmitted

Enteric Fecal-oral route Enteroviruses (polio, coxsackie B); noroviruses; rotaviruses (diarrhea)
Respiratory Respiratory or salivary route Influenza; measles; rhinoviruses (colds)
Zoonotic Vector (such as arthropods) Sandfly fever; dengue; West Nile encephalitis
Animal to human directly Rabies; cowpox
Sexually transmitted Sexual contact Herpes simplex virus type 2 (genital herpes); HIV

of other characteristics including viral shape and disease symp- MicroAssessment 13.1
toms are also considered.
Our discussion of classification will focus on the viruses Viruses consist of nucleic acid surrounded by a protein coat and
replicate only inside living cells. Viruses are typically icosahedral,
that infect animals. The names of virus families are derived helical, or complex. Most phages are naked, whereas animal viruses
from a variety of sources, but they all end in the suffix -viridae are either naked or enveloped. Viruses are classified based primarily
and are italicized. Their names follow no consistent pattern. In on the characteristics of their genome, such as type of nucleic
some cases, the name indicates the appearance of the viruses in acid and strandedness. Viruses are often grouped by their route of
the family—for example, Coronaviridae coming from corona, transmission.
which means “crown.” In other cases, the virus family is named 1. How are enveloped viruses different from naked viruses?
for the geographic area from which a member was first isolated. 2. List three ways in which viruses can be transmitted from one
Bunyaviridae is derived from Bunyamwera, a locality in Uganda, organism to another.
Africa. Each family contains numerous genera whose names 3. Most enteric viruses are naked. Why would this be so? +
end in -virus, making it a single word—for example, Enterovirus.
The species name is often the name of the disease the virus
causes—for example, poliovirus causes poliomyelitis. In contrast
to bacterial nomenclature, in which an organism is referred to by 13.2 ■ Bacteriophages
its genus and species name, viruses are commonly referred to only
by their species name. Formal scientific names are italicized, mak- Learning Outcomes
ing them easy to recognize. 3. Compare and contrast lytic, temperate, and filamentous phage
Although viruses have formal names, virologists most often infections.
refer to them by informal names that are not capitalized. In addi- 4. Describe two consequences of lysogeny.
tion, informal terms are often used to refer to groups of animal
viruses that are not taxonomically related but share critical char- An enormous variety of different phages exist, each with a char-
acteristics such as the primary route of transmission (table 13.2). acteristic shape, size, genome structure, and replication strategy.
Viruses transmitted via the fecal-oral route, for instance, are Although each is interesting, three general types stand out in their
referred to as enteric viruses, and viruses transmitted through the different relationships with their hosts (figure 13.4).
respiratory route are called respiratory viruses. Zoonotic viruses
cause zoonoses, which are diseases transmitted from an animal
to a human. One group of viruses, the arboviruses (meaning
Lytic Phage Infections:
arthropod borne), is so named because they are spread by arthro- T4 Phage as a Model
pods such as mosquitoes, ticks, and sandflies. The arthropods By definition, lytic or virulent phages exit the host at the end of
are biological vectors; when an infected arthropod takes a blood the infection cycle by lysing the cell. These viral infections result
meal from an animal, it transmits the virus. In many cases, these in the formation of new virus particles and are called productive
viruses can infect widely different species. The same arthropod infections. An intensively studied lytic phage is T4, a double-
might bite birds, reptiles, and mammals and transfer viruses stranded DNA phage. Its infection cycle illustrates how a phage takes
among those different groups. Arboviruses cause important over a cell, directing that cell’s activities solely to the synthesis of
diseases such as West Nile encephalitis, La Crosse encephalitis, new phage particles. The infection cycle, which is similar to that of
yellow fever, and dengue fever. biological vector, p. 442 West other lytic phages, can be viewed as a five-step process (figure 13.5).
Nile encephalitis, p. 655 La Crosse encephalitis, p. 655 yellow fever,
p. 682 dengue fever, p. 683 Attachment
In liquid, phage particles collide with their host cells by chance.
MicroByte Upon contact, a phage particle attaches by means of a protein on
An estimated 1031 bacteriophages exist on earth, the most numerous
its tail to a receptor on the cell surface or to an appendage such as
of all biological entities.
a pilus. In the case of T4, the receptor is on the bacterial cell wall.
 Part II The Microbial World 311
Focus Figure
1 Attachment
Virion

Infection
Phage attaches to
specific receptors on
the E. coli cell wall.
Host cell

Genetic alteration
Disease of host cell of host cell

2 Genome Entry
try
ry
y
Productive Infection Latent State
More virus produced Virus nucleic acid The tail contracts and
integrates into host phage DNA is injected
genome or replicates into the bacterial cell,
as a plasmid. leaving the phage
Release of virions— Release of virions— coat outside.
host cells lyse. host cells do not lyse.

Host cell multiplies— Host cell multiplies 3 Synthesis

Host cell dies. continuous release but phenotype often Phage genome is trans-
of virions. changed because cribed and phage proteins
of viral genes. synthesized. Phage DNA
is replicated, other virion
components are made,
FIGURE 13.4 Major Types of Relationships Between and host DNA is degraded.
Viruses and the Cells They Infect
? Which type of interaction is least harmful to the host?

4 Assembly

Phage components are

The receptors used by phages normally perform important func- assembled into mature
tions for the cell—the phages merely exploit the molecules for virions.
their own use. Cells that lack the receptor used by a given phage
are resistant to infection by that specific phage. DNA
Empty
inside
head
Genome Entry head
+
Following attachment, a bacteriophage injects its genome into the
+ +
cell. T4 does this by degrading a small portion of the bacterial cell
wall, using an enzyme located in the tip of its tail. This enzyme,
T4 lysozyme, differs from the lysozyme found in tears and eggs,
but it serves the same function—to degrade peptidoglycan. The
tail contracts so that the phage particle appears to “squat” on the 5 Release
surface of the cell. This action injects the phage DNA through the
host’s cell wall and membrane, and into the interior of the cell. The bacterial cell lyses
and many new infectious
The capsid remains on the outside of the cell. The separation of virions are released.
the nucleic acid from its protein coat prior to replication is a fea-
ture of all viruses. lysozyme, p. 62

MicroByte
The nucleic acid is so tightly packed inside the capsid that the
internal pressure is 10 times higher than that in a champagne bottle! FIGURE 13.5 Steps in the Replication of the Lytic Phage
T4 in E. coli
Synthesis of Phage Proteins and Genome ? If one phage particle infects one bacterial cell, how many particles
Within minutes of entry into a host cell, some T4 genes are are inside the cell 5 minutes after infection? Explain.
transcribed and translated. Thus, the cell begins synthesizing T4
proteins using host ribosomes. Not all phage-encoded proteins include (1) a nuclease that degrades the host cell’s DNA, and
are synthesized simultaneously, however. Rather, they are made (2) proteins that modify a subunit of the host cell’s RNA poly-
in a sequential manner during the course of infection. The first merase so that it no longer recognizes bacterial promoters. As a
viral proteins produced are referred to as early proteins, and are result, soon after infection, no host genes are expressed. In this
important for the initial steps of phage multiplication. These way, the phage takes over the metabolism of the bacterial cell for
312 Chapter 13 Viruses, Viroids, and Prions

its own purpose, namely the synthesis of more phage particles. Release
Bacterial enzymes already present in the cell, however, continue Late in infection, the phage-encoded enzyme lysozyme is pro-
to function. These are important to bacteriophage replication duced. This enzyme digests the host cell wall from within, caus-
because they allow biosynthesis and energy harvest to continue. ing the cell to lyse, thereby releasing phage. In the case of the T4
Towards the end of the infection cycle, the structural proteins phage, the burst size—the number of phage particles released—is
that make up the phage—including those that make up the capsid about 200. These phage particles then infect any susceptible
and tail—are synthesized. These are referred to as late proteins. cells in the environment, and the process of phage replication is
promoters, p. 168 repeated. The entire process from entry of the phage nucleic acid
to the exit of the phage takes about 30 minutes.
Assembly (Maturation)
Once multiple copies of the phage genome and the various struc-
tural components of the phage are produced, they assemble to Temperate Phage Infections:
produce new phage particles. This is a complex, multistep process. Lambda Phage as a Model
Once the phage head is formed, it is packed with DNA; the tail Temperate phages have the option of either directing a lytic
is then attached, followed by the addition of the tail spikes. Some infection (productive infection) or incorporating their DNA into
of these components self-assemble, meaning that the proteins join the host cell genome (figure 13.6). The latter situation is called a
together spontaneously to form a specific structure. In other steps, lysogenic infection, and the infected cell is a lysogen.
certain phage proteins serve as scaffolds on which various protein When a bacterial culture is infected with a temperate phage, some
components associate. The scaffolds themselves do not become a of the phage will enter the lytic cycle, whereas others will lysogenize
part of the final structure, much as scaffolding required to build a their host. Which cycle occurs is largely random, but the metabolic
house does not become part of the structure. state of the host cell has an influence. For example, if a bacterial cell
is growing slowly because of nutrient deprivation, then a lysogenic
infection is more likely to occur.
The most thoroughly studied temperate phage is lambda (λ).
Phage λ attaches This phage has a linear chromosome, but the two ends have com-
to bacterium. plementary single-stranded overhangs that join together inside
of the host cell to form a circular molecule. That molecule
can either direct a lytic infection or integrate into the
E. coli chromosome; lysogeny occurs when the
Injected linear phage molecule integrates. The integration process
DNA circularizes Prophage is integrated into uses a phage-encoded enzyme called an
and enters lytic or the bacterial chromosome.
lysogenic cycle. Integrated DNA integrase that inserts the phage DNA
into the host cell chromosome at
To lysogenic infection
specific sites, a process called site-
specific recombination. The inte-
Cell division.
grated phage DNA, a prophage,
replicates along with the host
To lytic chromosome prior to cell divi-
infection
sion. Although the prophage can
remain integrated indefinitely, it
To lytic infection
can also be excised from the host
chromosome by a phage-encoded
enzyme. When this happens a lytic
Replication of Excision of
phage DNA. infection begins.
phage DNA and
synthesis of phage- Whether the prophage persists or the
encoded proteins. lytic cycle begins depends on a complex series
of events involving phage-encoded regulatory pro-
teins, the study of which contributed to our current under-
standing of bacterial gene regulation. One of the proteins, a
repressor, prevents expression of the gene required for excision,
and is therefore essential for maintaining the lysogenic state.
Cells lyse, bacterial gene regulation, p. 179 repressor, p. 180
releasing new phage.

FIGURE 13.6 Steps in the Replication of the Temperate Phage λ in E. coli

? When is it advantageous for a temperate phage to excise its DNA from the host chromosome?
 Part II The Microbial World 313

Consequences of Lysogeny
Although a lysogen is morphologically identical to an unin-
fected cell, lysogeny has consequences. These include immunity
Filamentous F pilus to superinfection (infection by the same phage) and lysogenic
phage Phage attaches to the
Phage F pilus of a bacterial conversion.
DNA cell and injects its Lysogens are protected against infection by the same
single-stranded DNA.
phage because the repressor that maintains the prophage in
the integrated state will also bind to the operator on incoming
phage DNA. The operator is a regulatory region that controls
the expression of the genes that direct a lytic infection. Thus,
if another λ phage injects its DNA into a λ lysogen, that DNA
Phage DNA replicates; will not be expressed. This phenomenon is called immunity
phage capsomeres are to superinfection. Note that this type of immunity is different
synthesized and from that of the human immune system, which is discussed in
embedded in the
host cell membrane. chapters 14 and 15. operator, p. 180
Lysogenic conversion is a change in the phenotype of a
lysogen as a consequence of the specific prophage it car-
ries. For example, only strains of Corynebacterium
diphtheriae that are lysogenic for a certain phage
Outside (β phage) synthesize the toxin that causes diphthe-
environment ria. Similarly, lysogenic strains of Streptococcus
Phage
pyogenes and Clostridium botulinum produce
Carrier cell Carrier cell
DNA toxins responsible for scarlet fever and botulism,
respectively. If a toxin is encoded exclusively
Phage nucleic acid gains its capsid Capsomeres
as it extrudes through the membrane.
by phage genes, then only bacterial strains that
The bacteria do not lyse. carry the prophage will synthesize the toxin.
Some examples of lysogenic conversion are given
FIGURE 13.7 Replication of the Filamentous Phage
in table 13.3. diphtheria, p. 490
? Why might infected cells grow more slowly than their
uninfected counterparts?
Filamentous Phages:
M13 Phage as a Model
Filamentous phages are single-stranded DNA phages that look
Under ordinary growth conditions, the phage DNA is excised like long fibers. They cause productive infections, but the process
from the chromosome only about once in 10,000 divisions of the does not kill the host cells. Infected cells, however, grow more
lysogen. If, however, a lysogenic culture is treated with a DNA- slowly than their uninfected counterparts.
damaging agent such as ultraviolet light, the SOS repair system M13 is a filamentous phage that initiates infection by attach-
comes into play. This system activates a protease that destroys the ing to a protein on the F pilus of E. coli (figure 13.7). The single-
repressor protein responsible for maintaining the integration of stranded DNA phage genome then enters the cytoplasm of the
the prophage. As a consequence, the prophage is excised from the bacterial cell, where a host cell DNA polymerase synthesizes the
chromosome—allowing the phage to enter the lytic cycle—and a complementary strand. This double-stranded DNA is referred to
productive infection results. This process, phage induction, lets the as the replicative form (RF). One strand of the RF is then used as a
phage escape from a damaged host, much like rats fleeing a sink- template to make mRNA as well as multiple copies of the phage’s
ing ship. ultraviolet light, p. 115 SOS repair, p. 196 protease, p. 150 single-stranded genome (figure 13.8).

TABLE 13.3 Some Properties Encoded by Prophage (Lysogenic Conversion)

Microorganism Medical Importance Property Encoded by Phage

Corynebacterium diphtheriae Causes diphtheria Diphtheria toxin

Clostridium botulinum Causes botulism Botulinum toxin
Escherichia coli O157:H7 Causes hemolytic uremic syndrome Shiga toxin
Streptococcus pyogenes Causes scarlet fever Streptococcal pyrogenic exotoxins (SPEs)
Salmonella enterica Causes food poisoning Modification of lipopolysaccharide of outer membrane
Vibrio cholerae Causes cholera Cholera toxin
314 Chapter 13 Viruses, Viroids, and Prions

13.3 ■ The Roles of

ssDNA (+) strand Bacteriophages
in Horizontal
Host enzyme synthesizes
complementary strand. Gene Transfer
Learning Outcome
dsDNA (+
–) strand (RF)
5. Compare and contrast generalized and specialized
transduction.
Replication
Phages play important roles in horizontal gene transfer. As briefly
discussed in chapter 8, phages can transfer DNA from one bacte-
(+) strand DNA (–) strand DNA rial cell (the donor) to another (the recipient) in the process called
functions as transcribed transduction. There are two types of transduction: generalized and
phage genome into mRNA
specialized. horizontal gene transfer, p. 200

mRNA translated
into phage Generalized Transduction
coat protein
Generalized transduction results from a packaging error dur-
ing phage assembly. Some phages degrade the bacterial chro-
mosome into many fragments during lytic infection. Any of
Virion these short DNA fragments can be mistakenly packaged into
the phage head during assembly (see figure 8.20). Phage heads
that contain only bacterial genes in place of phage genes can-
FIGURE 13.8 Macromolecule Synthesis in Filamentous not direct a phage replication cycle. Because of this, they are
Phage Replication called generalized transducing particles rather than phage
? Which host enzyme synthesizes the complement to the ssDNA to particles. generalized transduction, p. 205
make the RF? Following its release from the phage-infected host, the
generalized transducing particle binds to another bacterial cell
M13 particles are assembled as the phage DNA is extruded and injects its DNA. That DNA may then integrate into the
from the cell. In this process, phage coat protein molecules are recipient cell by homologous recombination, replacing DNA
inserted into the host cell’s cytoplasmic membrane. At the same of the host cell. Any gene of the donor cell can be transferred
time, other phage-encoded proteins form pores that span the cyto- this way, which is why this mechanism is called generalized
plasmic and outer membrane. Then, as phage DNA is secreted transduction. homologous recombination, p. 201

through the pores, the coat protein molecules coat the single-
stranded DNA to form the nucleocapsids.
M13 phage is useful in certain recombinant DNA procedures Specialized Transduction
because its replication cycle produces both single- and double- Specialized transduction results from an excision mistake made
stranded DNA. If a researcher needs a preparation of a specific by a temperate phage during its transition from a lysogenic to
sequence of single-stranded DNA without its complement, the a lytic cycle. Following induction, the phage DNA is usually
DNA of interest can be cloned into an RF molecule of M13. When excised precisely from the bacterial chromosome. On rare occa-
the recombinant DNA is transformed into an E. coli cell, it will sions, however, a short piece of bacterial DNA flanking the phage
direct a productive phage infection, generating mature phage par- DNA is taken, and a piece of phage DNA remains in the bacterial
ticles containing only single-stranded recombinant DNA. chromosome (figure 13.9).
The excised DNA—containing both bacterial and phage
MicroAssessment 13.2 genes—replicates and then becomes incorporated into phage
heads during assembly. These phage particles do not carry the
Lytic phages lyse their host cells, whereas temperate phages either
lyse their host or integrate their DNA into the host cell’s genome. entire set of phage genes, so they are defective. The defective
Integrated viral DNA often codes for gene products that confer new phage particles are then released as the host cells lyse. When
properties on the host. Filamentous single-stranded DNA phages are a defective phage injects its DNA into another bacterial cell,
extruded from the host cell without killing the cell. both phage and bacterial DNA enter. The bacterial genes may
4. Describe two ways that phages can replicate without killing then integrate into the recipient’s genome via homologous
their host. recombination. In contrast to generalized transduction, only
5. Give an example of a virulent phage and of a temperate phage. bacterial genes adjacent to the integrated phage DNA can be
6. Why is lysogenic conversion medically important? + transferred, which is why the process is called specialized
transduction.
 Part II The Microbial World 315

MicroAssessment 13.3
1
Transduction is a process by which bacterial DNA is transferred from
one bacterium to another by a phage. Generalized transduction results
A temperate phage
from a DNA packaging error, whereas specialized transduction
injects its DNA into results from an error in excision of a prophage.
a bacterial host. 7. What is meant by a defective phage?
8. Which is more likely to be a specialized transducing phage—
a lytic or temperate phage?
9. Most temperate phages integrate into the host chromosome,
whereas some replicate as plasmids. Which kind of relationship
2 would you think would be more likely to maintain the phage in
The phage DNA integrates the host cell? Why? +
into the host cell DNA
to become a prophage.

13.4 ■ Bacterial Defenses

When the prophage is excised
Against Phages
3
from the bacterial chromosome,
a mistake is made; some bacterial Learning Outcome
DNA flanking the phage DNA is 6. Describe three mechanisms that confer bacterial resistance to
taken and a piece of the phage
phage infection.
DNA is left behind.

Bacteria have developed several defense mechanisms to resist

invasion by viruses. Without these, bacteria would not be able
4 to survive the ongoing war against phages. Defense mechanisms
Replication and assembly include altering receptor sites, restriction modification systems,
produces defective phage and the more recently discovered CRISPR system.
particles that carry certain
bacterial DNA in place of
some phage DNA. Preventing Phage Attachment
Recall that the first step in phage infection is attachment to specific
receptors on the cell surface. If a bacterium alters or covers a given
receptor, that cell becomes resistant to any phage that requires the
receptor. In some cases, the alteration has other benefits to the cell
5 as well. For example, Staphylococcus aureus produces a protein
The DNA of the defective phage (called protein A) that covers phage receptors on the cell surface.
is injected into the new host but That same protein also allows S. aureus to protect itself against cer-
cannot cause a productive
infection.
tain human host defenses. Surface polymers bacteria produce, such
as some used to form biofilms, may also cover phage receptors.

Restriction-Modification Systems
Restriction-modification systems protect bacteria from phage
6 The bacterial DNA integrates infection by quickly degrading incoming foreign DNA. They
into the host genome via
homologous recombination; do this through the combined action of two types of enzymes—
it can now be replicated along restriction enzymes and modification enzymes. Different bacteria
with the rest of the host DNA. have different versions of these enzymes, so there are hundreds
of varieties, each recognizing different sequences in DNA.
Integrated bacterial DNA The restriction enzyme recognizes specific short nucleotide
from original bacterial cell sequences and then cuts the DNA molecule at these sequences
(see figure 9.1). The modification enzyme protects the host cell
FIGURE 13.9 Specialized Transduction DNA from the action of the restriction enzyme. It does this by
? What mistake in the temperate phage replication cycle leads to adding methyl groups to the nucleobases recognized by the re-
specialized transduction? striction enzyme. Methylated sequences are invisible to the
restriction enzyme. Thus, a restriction enzyme will degrade
incoming phage DNA but not the host DNA. Occasionally, the
modification enzyme will methylate the incoming phage DNA
316 Chapter 13 Viruses, Viroids, and Prions

before the restriction enzyme has acted. When this happens, the Restriction enzyme Restriction enzyme does not
phage DNA will not be degraded, allowing it to replicate and lyse degrades unmethylated DNA degrade methylated DNA
the host cell. However, the DNA of the phage progeny will likely
be degraded by a restriction-modification system when it enters a Phage DNA Phage DNA
not methylated methylated
different bacterial strain (figure 13.10). restriction enzyme, p. 216 after entry
Restriction enzymes not only explain why some bacteria can
degrade foreign DNA, but they also played an important role in
the biotechnology revolution. The enzymes gave scientists a tool to
remove genes from one DNA molecule, and join them to another,
the basis for recombinant DNA technology. biotechnology, p. 215

CRISPR System Phage DNA degraded, Phage replication

no phage replication
The CRISPR system was more recently discovered, and details
about how it works are still being investigated. Bacterial cells
that survive some phage infections retain small segments of
phage DNA, incorporating them into the bacterial genome. The
segments of phage DNA—called spacer DNA—are inserted into
a chromosomal region called CRISPR, named for the character-
istic clusters of regularly interspersed short palindromic repeats
(figure 13.11). The spacer DNA provides a historical record of No phage

FIGURE 13.10 Restriction-Modification System

1 Infected bacterium
? How is the DNA modified by this system?

integration
Phage
of spacer
Cas protein Spacer

Phage DNA
Phage DNA
is cut into cas genes CRISPR array
fragments.

2 Surviving bacterium

Transcription

CRISPR RNA

RNA processing

3 Immunity

crRNAs

Phage
Cas-crRNA
complex

Invading viral
DNA is inactivated
by Cas-crRNA Cas-crRNA
complex. complex

FIGURE 13.11 CRISPR Defense System Against Phage Infection

? How does the CRISPR system target nucleic acid of an invading phage for destruction?
 Part II The Microbial World 317

past phage infections, allowing the bacterial cell as well as its

progeny to recognize and then block subsequent infections by
the same types of phages. How this works is not yet clear, but
the CRISPR system may function by a type of RNA interference.
The CRISPR array, including the spacer sequences, is transcribed
and then cut into small RNAs called crRNAs. These bind to pro- Bacteriophage
tein complexes called Cas (CRISPR-associated sequences). When plaques in lawn
the spacer RNA in a Cas base-pairs with nucleic acid of an invad- of bacterial cells
ing phage, that nucleic acid is targeted for destruction. In addition
to defending against phage, the CRISPR system can recognize
and destroy other entering foreign DNA that a bacterial strain has
encountered in the past. RNA interference, p. 182

MicroAssessment 13.4
Bacteria use several mechanisms to defend against viral infections. FIGURE 13.12 Phage Plaques
These include masking or altering phage receptor sites, and ? Why are plaques limited in size?
restriction-modification and CRISPR systems.
10. How do modification enzymes protect host cell DNA from
restriction enzymes?
MicroAssessment 13.5
11. How does a bacterial cell acquire a historical record of phage
infections? Bacteriophages require bacterial host cells. Plaque assays are used to
quantitate phage particles.
12. Which mechanisms that defend against phage infection would
also limit conjugation and DNA-mediated transformation? + 13. Explain what it is meant by plaque-forming units?
14. Is it important to have fewer phages than bacterial host cells
when doing a quantitative plaque assay? Explain. +

13.5 ■ Methods Used to Study

Bacteriophages 13.6 ■ Animal Virus Replication
Learning Outcome
Learning Outcomes
7. Describe how a plaque assay is used to detect and estimate the
8. Describe the steps of a generalized infection cycle of animal
numbers of phage particles.
viruses.
9. Compare and contrast the replication strategies of DNA viruses,
Viruses can multiply only inside living actively metabolizing
RNA viruses, and reverse-transcribing viruses.
cells, so studying viruses in the laboratory requires cultivating
appropriate host cells. It is considerably easier to grow bacterial
cells than animal cells, which is one reason why the study of bac- Understanding the infection cycle of animal viruses is particularly
teriophages advanced much more rapidly than investigations on important from a medical standpoint because virus replication
animal viruses. often depends on virally encoded enzymes, which offer potential
Plaque assays are routinely used to quantitate phage particles targets of antiviral drugs. By interfering with the activities of the
in samples such as sewage, seawater, and soil. In this assay, a enzymes required for viral replication, antiviral medications can
double layer of agar is used; the top layer, called soft agar, is inoc- slow the progression of a viral infection, often giving host defense
ulated with both a bacterial host and the phage-containing speci- systems enough time to clear the virus before symptoms appear.
men. It is then poured over the surface of an agar-filled Petri dish. As we discuss the infection cycles of animal viruses, it is
The bacteria present in the soft agar multiply rapidly, producing helpful to recall those described for bacteriophages, because they
a turbid layer or dense lawn of bacterial growth. Meanwhile, any have features in common. A generalized infection cycle of animal
phages in the specimen adsorb to susceptible bacteria and lyse viruses can be viewed as a five-step process.
them, releasing progeny phage that diffuse and infect neighbor-
ing bacteria. Plaques, circular zones of clearing, form in the lawn
due to cell lysis caused by the phage (figure 13.12). Each plaque Attachment
represents a plaque-forming unit (PFU) initiated by a single phage The process of attachment (adsorption) is basically the same in
particle infecting a cell. all virus-cell interactions. Animal virus surfaces are studded with
The plaque assay is done using different dilutions of the attachment proteins or spikes (see figure 13.2). The receptors to
phage suspension to ensure that the number of plaques on one which these proteins bind are usually glycoproteins located on the
plate can be accurately counted. The number is used to determine host cell plasma membrane, and often more than one receptor is
the titer, the concentration of infectious phage particles in the required for effective attachment. HIV, for example, must bind to
original phage suspension. two key molecules on the cell surface before it can enter the cell.
318 Chapter 13 Viruses, Viroids, and Prions

As with the receptors used by phages, the normal function of these the entire virion is taken into the cell. This differs from most
molecules is completely unrelated to their role in virus attachment. phages, where only nucleic acid enters and the capsid stays out-
glycoproteins, p. 29 HIV replication cycle, p. 700 side the bacterium.
Because a virion must bind to specific receptors, a particular Enveloped viruses enter the host cell by one of two mecha-
virus may be able to infect only a single or a limited number of nisms: fusion with the host membrane or endocytosis (figure 13.13).
cell types, and most viruses can infect only a single species. This In the case of fusion, the lipid envelope of the virion fuses with
accounts for the resistance that some animals have to certain dis- the plasma membrane of the host after the virion attaches to the
eases. For example, dogs do not contract measles from humans, host cell receptor, much as drops of oil in an aqueous medium can
nor do humans contract distemper from cats. Some viruses, such fuse together. As a result of fusion, the nucleocapsid is released
as the rabies virus, however, can infect unrelated animals such as directly into the cytoplasm. Viruses that enter by endocytosis
horses and humans with serious consequences in both. measles, exploit the process of receptor-mediated endocytosis, a normal
p. 537 rabies, p. 658 mechanism by which cells bring certain extracellular material
into the cell (see figure 3.48). The viral particles bind to
MicroByte the receptors that normally trigger and facilitate the process,
Tens to hundreds of thousands of receptors are present on each host causing the cell to take them up. As an example, the inter-
cell, allowing ample opportunity for a virion to encounter a receptor
action of the HIV envelope glycoproteins with host recep-
on a cell.
tors triggers entry of HIV into the host cell by endocytosis.
After a virion is taken into the cell, the viral envelope fuses with
the membrane of the endosome, releasing the nucleocapsid into
Penetration and Uncoating the cytoplasm. receptor-mediated endocytosis, p. 72
The mechanism an animal virus uses to enter its host cell depends Naked viruses, which have no lipid envelope, cannot fuse to
in part on whether the virion is enveloped or naked. In all cases, host membranes to enter cells. Therefore, the virions enter only

3 Nucleocapsid released
2 Membrane fusion into cytoplasm
1 Adsorption
Envelope of virion fuses Viral envelope remains part 4 Uncoating
Spikes of virion attach to with plasma membrane. of plasma membrane. Nucleic acid separates
specific host cell receptors.
from capsid.
Protein
spikes Fusion of virion and
host cell membrane
Envelope
Receptors
Nucleocapsid
Capsid
Host cell
plasma membrane Nucleic acid

(a) Entry by membrane fusion

2 Endocytosis
Plasma membrane surrounds
the virion, forming an endo-
cytic vesicle. 3 Release from vesicle
Envelope of virion fuses with
1 Adsorption 4 Uncoating
the endosomal membrane.
Attachment to receptors Nucleic acid separates
triggers endocytosis. from capsid.

(b) Entry by endocytosis

FIGURE 13.13 Entry of Enveloped Animal Viruses into Host Cells (a) Entry following membrane fusion. (b) Entry by endocytosis.
? Could phages enter bacteria by the process of membrane fusion? Why or why not.
 Part II The Microbial World 319

via endocytosis. Once in the endosome, the virus damages the

DNA viruses
vesicle membrane, resulting in the nucleocapsid being released
into the cytoplasm.
Following penetration, the virion is transferred to its site ss (–) ss (+)
of replication. Most DNA viruses multiply in the nucleus. They DNA DNA
enter the nucleus through nuclear pores using viral proteins that ds (+
–)
have nuclear localization signals. In all viruses, the nucleic acid DNA
ds (+
–) DNA ds (+
–) DNA
separates from its protein coat prior to the start of replication, the
process of uncoating. The protective coats are disassembled by
virus-specific mechanisms to release the viral genome and any ss (+) RNA ss (+) RNA ss (+) RNA
(mRNA) (mRNA) (mRNA)
accompanying proteins. This may occur simultaneously with
entry of the virion into the cell, or after the final intracellular des-
tination for viral replication has been reached. protein protein protein

(a)
Synthesis of Viral Proteins RNA viruses
and Replication of the Genome
ss (+)
As in the replication of phage, production of viral particles in RNA ss (–) RNA
an infected cell requires two distinct but interrelated events: (mRNA)
(1) expression of viral genes to produce structural and catalytic
proteins, such as capsid proteins and any enzymes required for
protein
replication; and (2) synthesis of multiple copies of the viral
genome. The fact that the relatively small viral genome is able
to direct production of more viral particles given the constraints ss (–) ss (+) RNA
posed by eukaryotic cells’ strict control of DNA replication and RNA (mRNA)
gene expression is a remarkable feat, and an area of intensive
research. protein
The replication strategy of viruses can be divided into three
general categories: those used by (1) DNA viruses, (2) RNA
viruses, and (3) reverse transcribing viruses (figure 13.14). As ds (+
–) ss (+) RNA
RNA (mRNA)
you will see in the following discussion, the type of genome has
a significant influence on the viral replication strategy, a pri-
protein
mary reason why virus classification takes genome structure into
account (see table 13.1).
(b)

Replication of DNA Viruses Reverse transcribing viruses

DNA viruses usually replicate in the nucleus of the host cell and ss (–) DNA ds (+
–) DNA
use the host cell machinery for DNA synthesis as well as gene
expression. These viruses often encode their own DNA poly- ss (+)
merase, however, thereby freeing them from dependency on cer- RNA
tain host cell constraints, such as replicating only when the host is (mRNA)
actively synthesizing its chromosome in preparation for division.
Poxviruses are an exception to the rule that DNA viruses replicate protein
in the nucleus. These large viruses replicate in the cytoplasm, and
encode all of the enzymes and factors necessary for DNA and (c)
RNA synthesis. FIGURE 13.14 Viral Replication Strategies Viral genomes are
Replication of double-stranded DNA viruses is fairly simple shown in black hexagons. The steps indicated by black arrows use
because they follow the central dogma of molecular biology (see host enzymes, while those indicated by red arrows use virally encoded
figure 7.1). The viral DNA can be replicated and expressed using enzymes.
the general mechanisms discussed in chapter 7. central dogma of ? Why do RNA viruses need to encode an enzyme in order to
molecular biology, p. 162 replicate their genome, while DNA viruses do not?
Replication of single-stranded DNA viruses is quite simi-
lar to that of their double-stranded counterparts, except that
the complement to the single-stranded molecule must first be (+) strand. Therefore, if the genome of a single-stranded DNA
synthesized. To understand this, recall that during DNA syn- virus is the (+) strand, the only way to produce identical copies
thesis, a (+) strand is used as a template to make the (–) strand, is to first use the genome as a template to make a (–) strand.
and, likewise, the (–) strand serves as a template to make the Then that (–) strand can serve as a template to make multiple
320 Chapter 13 Viruses, Viroids, and Prions

copies of the (+) strand. Although this might seem complex, of a segmented virus infect the same cell, reassortment can occur,
the host cell machinery can readily synthesize complementary meaning the viral particles that exit the cell can have various
DNA from a single-stranded template, making the replication combinations of segments from the initial infecting strains. For
of single-stranded DNA viruses relatively straightforward. example, if a virus has seven segments, numbered one through
seven, then the progeny must also have segments numbered one
Replication of RNA Viruses through seven, but the segments can originate from either of the
The vast majority of RNA viruses are single-stranded, and they two parent strains. Influenza A virus is an example of a segmented
replicate in the cytoplasm. Their replication always requires a virus that undergoes reassortment, resulting in a phenomenon
virally encoded RNA polymerase, often called a replicase. The called antigenic shift. Antigenic shift and drift will be discussed
need for a virally encoded enzyme becomes obvious when you in detail in chapter 21. antigenic shift, p. 511

recall that the RNA polymerase used in transcription requires

DNA as a template to make a complementary copy of RNA; it Replication of Reverse-Transcribing Viruses
cannot synthesize RNA from an RNA template. RNA viruses, Reverse-transcribing viruses encode the enzyme reverse
however, must direct synthesis of a complementary copy of RNA transcriptase, which synthesizes DNA from an RNA template.
from an RNA template. In other words, the virally encoded repli- This runs counter to the central dogma of molecular biology
case must be an RNA-dependent RNA polymerase. in that an RNA molecule is used as a template to make DNA.
Replicases lack proofreading ability, and therefore make central dogma of molecular biology, p. 162
more mistakes than DNA polymerases. Because of the relatively Retroviruses, which include the human immunodeficiency
high number of mutations generated, some RNA viruses can adapt virus (HIV), have a (+) strand RNA genome, and carry reverse
more quickly to selective pressures than their DNA counterparts. transcriptase within the virion. After entering the host cell, the
Influenza viruses, for example, exhibit a type of antigenic varia- reverse transcriptase uses the RNA genome as a template to make
tion called antigenic drift. This occurs as mutations accumulate one strand of DNA. The complement to that strand is then syn-
in genes encoding key viral surface proteins that are recognized thesized to make double-stranded DNA, which integrates into the
by the immune system. Because of such changes, a person whose host cell chromosome. Once integrated, the genome can direct a
immune response protected him or her against influenza virus one productive infection, or remain in a latent state. This latent state
year may not be protected against the variant that circulates the is similar to what occurs with phage λ. Once the DNA copy is
next year. antigenic drift p. 510 made, however, it cannot be eliminated from the cell. replication
Some single-stranded RNA viruses are (+) strand, meaning of HIV, p. 699
their genome functions as mRNA. Once introduced into a cell, Hepatitis B virus (HBV) has a complicated infection cycle
the genome is translated to produce a replicase. This enzyme that involves a temporary RNA copy of its double-stranded DNA
then makes multiple copies of the complement, a (–) strand, from genome. When the HBV genome enters a cell, it is transcribed
which more copies of the (+) strand can be made. Each (+) strand by the cell’s machinery to produce RNA. That molecule codes
RNA molecule can be translated to make more viral proteins. for viral proteins, and also serves as a temporary genome copy
In addition, the (+) strand molecules can be packaged to make called pregenomic RNA (pgRNA). During the assembly process,
nucleocapsids during the assembly process. the pgRNA along with reverse transcriptase is packaged into the
A more complicated situation arises for (–) single-stranded maturing viral particles. There, the reverse transcriptase functions
RNA viruses. In this case, the RNA genome is the complement of to make DNA using the RNA as a template. DNA synthesis stops
mRNA and cannot be translated. As a result, replicase cannot be once the virion buds from the membrane, resulting in a DNA
made immediately. To solve this problem, the virus carries repli- molecule that is not fully double-stranded. replication of hepatitis
case within the viral particle so that when the genome enters the B virus, p. 600
cell, the accompanying replicase can immediately produce (+)
strand RNA using the (–) strand RNA genome as a template. The
(+) strand RNA is then translated to produce more replicase. Assembly
Some of the replicase molecules are packaged into the viral Viral assembly involves the same general principles described for
particles during the assembly process for use in the next infec- phages. The protein capsid must be formed, and then the genome
tion cycle. and any necessary enzymes packaged within it. In animal viruses,
Double-stranded RNA viruses, which are relatively uncom- this assembly process takes place in the nucleus or in the cyto-
mon, must also carry their own replicase because the host cell plasm, depending on the virus. Many viruses assemble near the
translation machinery is unable to translate double-stranded RNA. plasma membrane close to their site of release. Golgi apparatus
The replicase immediately uses the (–) strand of the double- p. 77 microtubules, p. 73
stranded molecule as a template to make (+) strand RNA. This
molecule is then translated to make more replicase, and the infec-
tion cycle can continue. Release
Some RNA viruses have segmented genomes, with different Just as the entry mechanism into a host cell depends on
segments containing different genes. All of the segments must be whether the virion is enveloped or naked, so does its release.
replicated for new virions to be produced. If two different strains Most enveloped viruses are released by budding, a process
 Part II The Microbial World 321

3 Nucleocapsid extrudes from the host cell,

becoming coated with matrix proteins
2 Viral matrix protein and envelope with protein spikes.
coats inside of
1 Viral proteins that will plasma membrane. 4 New virus is
become envelope spikes released.
insert into host plasma
membrane. Enveloped
virus
Viral proteins

Host plasma Matrix

membrane protein
Intact host
Nucleic acid membrane
Capsid

(a)

FIGURE 13.15 Mechanisms for Releasing Enveloped

Virions (a) Process of budding. (b) Electron micrograph of virions
budding from the surface of a human cell.
? What component of the virion is gained in the process of
budding?

process called apoptosis, or programmed cell death, prior to the

release of the virus particles. The immune response of the animal,
which is directed toward eliminating the virus, can also lead to the
same process. The virions released from the dead cells may then
invade any healthy cells in the vicinity. apoptosis, p. 350
To be maintained in nature, infectious virions must leave one
animal host and be transmitted to another. Viral particles may be
shed in feces, urine, genital secretions, blood, or mucus and saliva
released from the respiratory tract during coughing or sneezing.
(b)
From there, the virus enters the next host to begin another round
of infection.
whereby the virus acquires its envelope (figure 13.15). Before
MicroAssessment 13.6
budding occurs, virally encoded protein spikes insert into
specific regions of the host cell’s membrane. Matrix protein Animal viruses and bacteriophages share similar features in their
accumulates on the inside surface of those same regions. infection cycle. Because many animal viruses are enveloped and
phages are not, differences exist in their entry and exit. The genomes
Assembled nucleocapsids are then extruded from the cell of animal viruses are diverse, influencing viral replication strategies.
at these regions, becoming covered with a layer of matrix
15. Why are virally encoded enzymes medically important?
protein and lipid envelope in the process. Not all enveloped
viruses have plasma membrane–derived envelopes, however. 16. How do enveloped viruses exit a cell?
Some obtain their envelope from the membrane of an organ- 17. Why can viruses not replicate independently of living cells? +
elle such as the Golgi apparatus or the rough endoplasmic
reticulum. They do this by budding into the organelle. From
there, they are transported in vesicles to the outside of the cell. 13.7 ■ Categories of Animal
Budding may not destroy the cell because the membranes can Virus Infections
be repaired after the viral particles exit. rough endoplasmic
reticulum, p. 76
Learning Outcome
Naked viruses are released when the host cell dies. How this
10. Compare and contrast acute infections and the two types of
occurs, however, is sometimes quite different from how bacte- persistent infections caused by animal viruses.
riophages kill their hosts. Many viruses trigger a normal cellular
322 Chapter 13 Viruses, Viroids, and Prions

The relationship between viruses and

Acute infection (influenza)
their animal hosts can be divided into

infectious virions
State of Virus
two major categories of infections:

symptoms and
Appearance of
Infectious virions Virus disappears
acute and persistent (figure 13.16). Influenza Disease after disease ends.
Acute infections are characterized
by the sudden onset of symptoms of
a relatively short duration. In contrast,
persistent infections can continue for Time (days)
years, or even the life of the host with
or without symptoms. Although these (a)
can be compared to certain bacterio- Chronic infection (hepatitis B)
phage infections, the situation is con-

infectious virions
State of Virus

symptoms and
Appearance of
siderably more complicated because After initial infection with or
of the host defenses, which will be Hepatitis B Release of virus without disease symptoms,
discussed in later chapters. For now, infectious virus is released
from host with no symptoms.
simply realize that the immune system
is constantly working to detect and
destroy virus-infected cells. Viruses, Days Time Years
meanwhile, have mechanisms that
(b)
evade some of these defenses.
Although we often categorize Latent infection (cold sores)
viral infections as either acute or per-
infectious virions

State of Virus
symptoms and
Appearance of

sistent, they do not always fall neatly Cold Cold After initial infection, virus
into a single category. When a person sores Virus sores is maintained in neurons in
activation non-infectious state. Virus
is first infected with HIV, for instance, activated to produce new
the virus replicates to high levels, Non-infectious disease symptoms.
causing acute symptoms including
fever, fatigue, swollen lymph nodes, Days Time Years
and headache. The immune system (c)
soon eliminates most virions, and FIGURE 13.16 Types of Infection by Animal Viruses (a) Acute infection. (b) Persistent
the symptoms subside. However, the infection—chronic. (c) Persistent infection—latent.
DNA copy of the viral genome inte- ? Which type of infection is most likely to lead to immunity?
grates into the host cell chromosome,
resulting in a persistent infection with subsequent symptoms asso- an actual infection, however, with different cells experiencing dif-
ciated with acquired immunodeficiency syndrome (AIDS) devel- ferent types of infection.
oping after many years. Thus, HIV infection has features of acute Chronic infections are characterized by the continuous pro-
and persistent infections. duction of low levels of viral particles. In some cases, this is similar
to what occurs in filamentous bacteriophage infections with each
infected cell surviving and slowly releasing viral particles. In other
Acute Infections cases, the infected cell lyses, but only a small proportion of cells
On a cellular level, acute infections can be compared to productive is infected at any given time, resulting in a low number of viral
lytic infections by bacteriophages, resulting in a burst of virions particles being continuously released—for example, hepatitis B
being released from infected host cells. A critical difference, virus infection. From a practical standpoint, the important aspect
however, is that while the virus-infected cells often die, the host of either type of persistent infection is the continuous production of
may survive. This is because the immune system of the animal infectious viral particles, often in the absence of disease symptoms.
host may gradually eliminate the virus over a period of days to Consequently, a person can transmit the virus to others even in the
months. Examples of diseases due to acute viral infections include absence of symptoms. For example, some people infected with
influenza, mumps, and poliomyelitis. Symptoms of the diseases hepatitis B virus develop a chronic infection. They become carriers
result from localized or widespread tissue damage following cell of the virus, able to pass it to other people through blood and body
death as well as damage caused by the immune response itself. fluids. filamentous phage infection, p. 313 hepatitis B virus, p. 599
influenza, p. 508 mumps, p. 583 polio, p. 656 Latent infections are analogous to lysogeny by bacterio-
phages. The viral genome remains silent within a host cell, yet can
reactivate to direct a productive infection. Some viruses do not inte-
Persistent Infections grate into the host cell chromosome; rather, they replicate indepen-
Persistent infections remain for years, or even the life of the dently of the host genome, much like a plasmid. The silent viral
host—sometimes without any symptoms. In laboratory stud- genome is called a provirus. The fact that a provirus cannot be
ies, two general types of persistent infections can be identified: eliminated from the body means that the disease can recur even after
chronic and latent (table 13.4). These categories may overlap in an extended period without symptoms. This explains why cold
 Part II The Microbial World 323

TABLE 13.4 Examples of Persistent Infections

Virus Type of
Infection Cells Involved Disease

Hepatitis B virus Chronic Hepatocytes (liver cells) Hepatitis, cirrhosis, hepatocellular carcinoma
Hepatitis C virus Chronic Hepatocytes (liver cells) Hepatitis, cirrhosis, hepatocellular carcinoma
Herpes simplex virus type 1 Latent Neurons of sensory ganglia Primary oral herpes and recurrent herpes
simplex (cold sores)
Herpes simplex virus type 2 Latent Neurons of sensory ganglia Genital herpes and recurrent genital herpes
Varicella zoster (Herpesviridae family) Latent Satellite cells of sensory ganglia Chickenpox and herpes zoster (shingles)
Cytomegalovirus Latent Salivary glands, kidney CMV pneumonia, eye infections,
(CMV; Herpesviridae family) epithelium, leukocytes mononucleosis, congenital CMV infection
Epstein-Barr virus Latent B cells, which are involved Burkitt’s lymphoma
in antibody production
Human immunodeficiency virus (HIV) Chronic Activated helper T cells, macrophages AIDS
Latent Memory helper T cells

sores, which are caused by herpes simplex type 1 virus (HSV-1), MicroAssessment 13.7
can recur. HSV-1 causes an acute infection in mucosal epithelial
cells, leading to the typical symptoms of cold sores. From there, the Many viruses cause acute infections in which viruses multiply and
spread rapidly in the host. Some viruses cause persistent infections;
virus can spread to sensory nerve cells where it remains latent. these long-term infections can be chronic or latent.
Later, the latent virus can reactivate to cause another episode of cold
18. Distinguish between acute and persistent viral infections at the
sores (figure 13.17). What reactivates the virus is not clear, but
cellular level.
certain physiological or immunological changes in the host are
19. How are latent viral infections different from chronic ones?
often involved. herpes simplex type 1 virus, p. 582
20. Could the same type of virus cause both an acute and a
persistent infection? Explain. +
Latent Cranial nerve Pons
viral DNA

13.8 ■ Viruses and Human Tumors

Virus moves up
cranial nerve. Learning Outcome
11. Describe the roles of proto-oncogenes and tumor suppressor
genes in controlling cell growth, and how some viruses can
The initial infection in children causes
cold sores and sometimes sore throat. circumvent this control.
The virus then moves along a sensory Brain
cranial nerve to the cell body near the stem A tumor is an abnormal growth of tissue resulting from a malfunc-
brain, where it becomes latent.
tion in the normally highly regulated process of cell growth. Some
tumors are benign, meaning they do not metastasize (spread) or
(a) Virus becomes latent after initial infection invade nearby normal tissue. Others are cancerous or malignant,
meaning they have the potential to metastasize.
Control of cell growth involves genes that stimulate cell
Activation of
virus in neuron growth, called proto-oncogenes, and ones that inhibit cell growth,
termed tumor suppressor genes. These genes work together to
regulate growth and cell division. Mutations that result in inap-
Virions move down propriate timing or level of expression of proto-oncogenes or
cranial nerve. repression of tumor suppressor genes are the major cause of
abnormal and/or uncontrolled growth. A single change in the
DNA sequence of these regulatory genes is probably not enough
The latent virus is activated, moves
back along the sensory nerve to the
to cause a tumor; rather, multiple changes at different sites are
face and causes cold sores again. required. The changes can result from different factors, including
viruses. Some viruses, for example, carry genes called oncogenes,
(b) Activation of latent virus which are very similar in DNA sequence to proto-oncogenes.
Consequently, their entry into cells can interfere with the cell’s
FIGURE 13.17 Infection Cycle of Herpes Simplex Virus, HSV-1 own control mechanisms, leading to tumor formation. However,
? What term is used to describe a virus in its latent state? the majority of tumors are not caused by viruses but by mutations
324 Chapter 13 Viruses, Viroids, and Prions

TABLE 13.5 Viruses Associated with Cancers in Humans

Virus Type of Nucleic Acid Kind of Tumor

Human papillomaviruses (HPVs) DNA Different kinds of tumors, caused by different HPV types
Hepatitis B DNA Hepatocellular carcinoma
Epstein-Barr DNA Burkitt’s lymphoma; nasopharyngeal carcinoma; B-cell lymphoma
Hepatitis C RNA Hepatocellular carcinoma
Human herpesvirus type 8 DNA Kaposi’s sarcoma
HTLV-1 RNA (retrovirus) Adult T-cell leukemia (rare)

in host genes that regulate cell growth. Most virus-induced tumors MicroByte
are caused by certain DNA viruses (table 13.5). In 2010, a government advisory panel concluded that environmental
Fifteen human papillomaviruses (HPVs) are associated with pollutants are grossly underestimated as a cause of cancer.
the development of cancers. Although the mechanism by which
the viruses cause cancer is not entirely clear, several HPV-
MicroAssessment 13.8
encoded proteins appear to interfere with the function of an impor-
tant tumor suppressor gene product. The FDA approved a vaccine The majority of tumors are not caused by viruses but by mutations in
that contains two types of HPV that cause approximately 70% of certain host genes that regulate cell growth. The most common viral
cause of tumors are the DNA tumor viruses.
cervical cancers, along with two types of HPV that cause genital
warts. Other viruses implicated in cancer include human herpesvi- 21. Name three viruses that cause tumors in humans.
rus type 8 (HHV-8), which can cause Kaposi’s sarcoma; Epstein- 22. Explain why a vaccine can prevent cervical cancer.
Barr virus (EBV), which can cause Burkitt’s lymphoma; hepatitis 23. Why would it be advantageous to a virus to interfere with
B and C viruses, which can cause hepatocellular carcinoma the function of proto-oncogenes or tumor suppressor genes? +
(liver cancer); and T-cell lymphotrophic virus type 1 (HTLV-1),
which can cause adult T-cell leukemia (table 13.5). In all of these
cases, only a small percentage of infected individuals develop the 13.9 ■ Cultivating and Quantitating
cancer, indicating that other important factors must be involved. Animal Viruses
Epstein-Barr virus, p. 680 Kaposi’s sarcoma, p. 707 hepatitis B virus,
p. 599 papillomaviruses, p. 308 Learning Outcomes
The various effects that animal viruses can have on their host 12. Describe how animal cells are cultivated in the laboratory.
cells are shown in figure 13.18. 13. Describe the methods used to quantitate animal viruses.

FIGURE 13.18 Various Effects of Animal Viruses on the To study viruses, they must be grown in the laboratory in
Cells They Infect an appropriate host. This is much more difficult to do with
? What relationship is the most destructive to the infected cell? animal viruses than it is with phages.

Viral DNA
integrates into Viral DNA Virus replicates
host DNA. forms plasmid. in host.
Viral DNA
as a plasmid

or or or

Tumor Latent infection Tumor Latent infection Productive infection Productive infection
Normal cells are Viral DNA replicates Normal cells are Viral DNA replicates New virions released New virions released
transformed into as part of the transformed into as a plasmid without when host cell lyses. by budding.
tumor cells. chromosome without tumor cells. harming the host.
harming the host.
 Part II The Microbial World 325
Tissue

1 Cut tissue into small pieces

and incubate with a protease
(trypsin) to separate cells.
Single cells

2 Place cells into flask

with growth medium.

Monolayer
3 Allow cells to settle on bottom
of flask and grow into a single
layer (a monolayer).

100 μm
Stained cells
FIGURE 13.19 Preparation of Primary Cell Culture in monolayer

? Why must primary cell cultures be restarted every so often?

Cultivating Animal Viruses Effects of Viral Replication on Cell Cultures

Initially, the only way to study animal viruses was to inoculate Many viruses can be detected by their effect on cells in cell cul-
live animals with a suspension of virus particles. Later, embryo- tures. A virus propagated in tissue culture often causes distinct
nated (fertilized) chicken eggs were used to grow viruses, which morphological alterations in infected cells, called a cytopathic
made research much easier. Although improved techniques have effect (figure 13.20). The host cells, for example, may change
largely replaced these methods, they are still used to cultivate cer-
tain viruses. For example, influenza viruses are grown in embryo-
nated chicken eggs to manufacture vaccines against influenza.
vaccine, p. 421 influenza virus, p. 508
Today, cell culture or tissue culture is commonly used to
cultivate most animal viruses. Animal cells—grown in a liquid
medium contained in special screw-capped flasks—are used as
host cells for the virus culture. Much like bacteria, animal cells
bathed in the proper nutrients can divide repeatedly. Animal cells
grow much more slowly than most bacteria, however, dividing
no faster than once every 24 hours (E. coli can divide every 20
minutes). Animal cell types that make up solid tissues typically
grow as a monolayer, a single sheet of cells adhering to the bottom
of the flask. Cells from blood grow as single cells in suspension.
Although cell culture is generally easier than growing viruses in (a) 0.5 μm
living animals, not all viruses can be grown in this way.

Obtaining and Maintaining Cell Cultures

One way to obtain animal cells for culture is to remove tissue from an
animal, and then process it to get individual cells. These cells can then
be grown in a Petri dish with a liquid nutrient medium. Cells acquired
in this way form primary cultures (figure 13.19). One problem with Dead cells
this approach is that normal cells can divide only a limited number of
times, even when diluted into a fresh medium, and so new primary
cultures must regularly be made. To avoid this problem, tumor cells
are often used in cell culture. These multiply indefinitely in vitro,
resulting in what is called an established cell line.
(b) 0.5 μm
MicroByte
The commonly used HeLa cell line was derived from cervical cancer FIGURE 13.20 Cytopathic Effects of Virus Infection on
cells taken from a patient named Henrietta Lacks, who eventually Tissue Culture (a) Fetal tonsil diploid fibroblasts growing as a
died from the cancer. monolayer, uninfected. (b) Same cells infected with herpes simplex
virus.
326 Chapter 13 Viruses, Viroids, and Prions

shape, detach from the surface, or lyse. Infected cells may fuse
into a giant multinuclear cell (syncytium), a mechanism of viral
spread. Several viruses, such as HIV and measles, cause this cyto-
pathic effect.
Certain viruses cause an infected cell to form a distinct region
called an inclusion body, the site of viral replication. The posi-
tion of an inclusion body in a cell depends on the type of virus.
Because cytopathic effects indicate that a particular cell is infected Red blood Virions Hemagglutination
cells
and they are often characteristic for a particular virus, they are
(a)
useful to scientists studying and identifying viruses. Controls

Dilution
Quantitating Animal Viruses

1/1024
1/256
1/128

1/512

Neg.
Pos.
1/32

1/64
1/16
1/2

1/4

1/8
Patient Titer
One of the most precise methods for determining the concen-
tration of animal viruses in a sample is the plaque assay. This A 256
is similar in principle to the method described for quantitating B 32
bacteriophages, but in the case of animal viruses, a monolayer of
C 512
tissue culture cells is the host. Again, clear zones surrounded by
uninfected cells are counted to determine the viral titer. D 8
If a sample contains a high enough concentration of viruses E 32
to be seen with an electron microscope, direct counts can be
used to determine the number of viral particles in a suspension F 128
(figure 13.21). G 64
A viral titer can be estimated using a quantal assay. In this
H >2
method, several dilutions of the virus preparation are administered
to a number of animals, cells, or chick embryos, depending on the Hemagglutination No
host specificity of the virus. The titer of the virus, or the endpoint, (b) hemagglutination
is the dilution at which 50% of the inoculated hosts are infected
FIGURE 13.22 Hemagglutination (a) Diagram showing virions
or killed. This can be reported as either the ID50 (infective dose), combining with red blood cells, resulting in hemagglutination.
or the LD50, (lethal dose). (b) Assay of viral titer. Red blood cells normally form a pellet when they
Certain viruses cause red blood cells to agglutinate (clump). sink to the bottom of the wells, whereas agglutinated ones do not.
This phenomenon, hemagglutination, occurs when individual ? Which patient has the highest titer of antibodies?
viral particles attach to surface molecules of multiple red blood
cells simultaneously, connecting the cells to form an aggregate
(figure 13.22). Hemagglutination is visible only at high concen- agglutination is the titer of the virus. One group of animal viruses
trations of viruses, and can be used to determine only the relative that can agglutinate red blood cells is the orthomyxoviruses, of
concentration of viral particles. Hemagglutination is measured which the influenza virus is a member.
by mixing serial dilutions of the viral suspension with a standard
amount of red blood cells. The highest dilution showing maximum MicroAssessment 13.9
Various hosts are required to grow different viruses. These include
whole animals, embryonated chicken eggs, and cell cultures. Viruses
that lyse their host cells can be assayed by counting plaques. Other
methods include direct counts, quantal assays, and hemagglutination.
24. In tissue culture, why is it advantageous to use tumor cells
Empty capsid
rather than normal cells?
25. Discuss two methods used to quantitate animal viruses.
26. How might syncytia formation of infected cells benefit
viruses? +

13.10 ■ Plant Viruses

Learning Outcomes
100 nm 14. Compare and contrast the mechanisms by which plant and animal
viruses enter host cells.
FIGURE 13.21 Electron Micrograph of Calicivirus Note that
empty capsids are readily distinguishable. 15. Discuss the ways that plant viruses can be transmitted to their
hosts.
? Is a virion with an empty capsid infectious?
 Part II The Microbial World 327

Viral diseases of plants are economically important, particularly when

they occur in crop plants such as corn, wheat, rice, soybeans and sugar
beets. Over half of the crop yields can be lost during a serious virus
infection. Viral diseases are especially common among perennial
plants (those that live for many seasons), such as tulips and potatoes,
and those propagated vegetatively (not by seeds), such as potatoes.
Viral infection of plants can be recognized through vari-
ous outward signs, including yellowing of foliage with irregular
lines appearing on the leaves and fruits (figure 13.23). Individual
cells or specialized organs of the plant may die, and tumors may
appear. Usually, infected plants become stunted in their growth,
although in a few cases growth is stimulated, leading to deformed
structures. Plants generally do not recover from viral infections,
(a) because unlike animals, plants are not capable of developing spe-
cific immunity to rid themselves of invading viruses. In severely
infected plants, virions may accumulate in enormous quantities.
For example, as much as 10% of the dry weight of a tobacco
mosaic virus–infected plant may consist of virus. The virus, which
causes a serious disease of tobacco, retains its infectivity for at
least 50 years, which explains why it is usually difficult to eradi-
cate the virions from a contaminated area. Other plant viruses are
also extraordinarily stable in the environment.
In a few instances, plants have been purposely maintained
in a virus-infected state. The most well-known example involves
tulips, in which a virus transmitted through the bulbs can cause
a desirable color variegation of the flowers (figure 13.24). The
infecting virus was transmitted through bulbs for a long time
before the cause of the variegation was even suspected.
In contrast to phages and animal viruses, when plant viruses
infect a cell they do not attach to specific receptors. Instead, they
(b)
enter through wound sites in the cell wall, which is otherwise very
tough and rigid. Infection in the plant can then spread from cell to
cell through openings (the plasmodesmata) that interconnect cells.
Plant viruses can be transmitted through soil contaminated
by prior growth of infected plants, and by growers themselves.

(c)

FIGURE 13.23 Signs of Viral Diseases of Plants (a) A healthy

wheat leaf can be seen in the center. The yellowed leaves on either
side are infected with wheat mosaic virus. (b) Typical ring lesions
on a tobacco leaf resulting from infection by tobacco mosaic virus.
(c) Stunted growth (right) in a wheat plant caused by wheat mosaic FIGURE 13.24 Tulips with Symptoms Resulting from Virus
virus. Infection
? How do most plant viruses enter plants? ? What other symptoms can plant viruses cause?
328 Chapter 13 Viruses, Viroids, and Prions

A small percentage of the known plant viruses are transmitted

through contaminated seeds, tubers, or pollen. Virus infections
can also spread through grafting of healthy plant tissue onto
diseased plants. Tobacco mosaic virus is transmitted to healthy
seedlings on the hands of workers who have been in contact with
the virus from infected plants. The most important transmitters of
plant viruses are probably insects; thus, insect control is a critical
tool for preventing the spread of plant viruses.

MicroByte
Tobacco in cigarettes, chewing tobacco, and cigars may carry the
tobacco mosaic virus. Smokers should wash their hands thoroughly
before handling plants.

MicroAssessment 13.10
Plant viruses cause many plant diseases and are of major economic
importance. They invade through wound sites in the cell wall and are
spread through soil, insects, and growers.
27. How does a plant virus penetrate the tough outer coat of
the plant cell?
28. How are plant viruses transmitted?
29. Why is it especially important for plant viruses to be
stable outside the plant? + FIGURE 13.25 Electron Micrographs of Viroids
? Will heating affect the structure of a viroid?

13.11 ■ Other Infectious Agents: Prions

Viroids and Prions Prions are composed solely of protein, which is reflected in
the name (derived from proteinaceous infectious agent). These
Learning Outcomes agents have been linked to a number of slow, always fatal, human
16. Describe the chemical structure of viroids and prions. diseases including Creutzfeldt-Jakob disease and kuru, as well
17. Compare hosts of viroids and prions. as to animal diseases such as scrapie (sheep and goats), bovine
18. Describe the process by which prions accumulate in tissues. spongiform encephalopathy (cattle), and chronic wasting disease
(deer and elk) (table 13.6). In all these diseases, prion proteins
Although viruses are composed of only nucleic acid surrounded
by a protective protein coat, other infectious agents are even sim-
pler in structure. These are the viroids and prions. TABLE 13.6 Prion Diseases
Disease Host
Viroids Scrapie Sheep and goats
Viroids consist solely of a single-stranded RNA molecule that var- Bovine spongiform Cattle
ies in size from 246 to 375 nucleotides and forms a closed ring encephalopathy
(figure 13.25). This is about one-tenth the size of the smallest infec- Chronic wasting disease Deer and elk
tious viral RNA genome known. A great deal of hydrogen bonding Transmissible mink Ranched mink
exists between complementary bases in the viroid RNA so that the encephalopathy
single-stranded molecule appears to be a double-stranded structure. Exotic ungulate Antelope in South Africa
All viroids that have been identified infect only plants, where encephalopathy
they cause serious diseases including potato spindle tuber, chry- Feline spongiform Cats
santhemum stunt, citrus exocortis, cucumber pale fruit, hopstunt, encephalopathy
and cadang-cadang. Like plant viruses, they enter plants through Kuru Humans (caused by cannibalism)
wound sites rather than binding to specific receptors. A great deal Variant Creutzfeldt- Humans (caused by consumption
is known about the structure of viroid RNA, but many questions Jakob disease of prion-contaminated beef)
remain. How do viroids replicate? How do they cause disease? Creutzfeldt-Jakob disease Humans (inherited)
How did they originate? Do they have counterparts in animals, or
Gerstmann–Straussler Humans (inherited)
are they restricted to plants? The answers to these questions will Scheinker syndrome
provide insights into new and fascinating features of this unusual
Fatal familial insomnia Humans (inherited)
member of the microbial world.
 Part II The Microbial World 329

accumulate in neural tissue. For unknown reasons, neurons die Prions are unusually resistant to heat and chemical treatments that
and brain function deteriorates as the tissues develop characteris- are commonly used to inactivate infectious agents.
tic holes (figure 13.26). The characteristic spongelike appearance Prions violate the central dogma of replication that requires
of the brain tissues gave rise to the general term transmissible nucleic acid act as a template for replication of macromolecules.
spongiform encephalopathies, which refers to all prion diseases. Prions are not made up of nucleic acid, making their accumulation
prions, p. 664 inside cells difficult to explain. It is hypothesized that PrPSC inter-
Considering that prions lack any nucleic acid, how they acts directly with PrPC and converts its folding properties from
accumulate in tissue and cause disease has long been an intrigu- PrPC to PrPSC (figure 13.27). This conversion may require another
ing question. An answer began to emerge when it was discovered unknown factor. As a result, cells that continue to produce PrPC
that uninfected animals synthesize a protein in neurons, especially accumulate PrPSC. How prions cause neurons to die is not known.
in the brain, that has an identical or nearly identical amino acid In most cases, the disease is transmitted only to members
sequence to infectious prions. Prion proteins from different ani- of the same species. However, the barrier to prion transmission
mals are similar but not identical. A key difference between the between species also depends on the strain of prion. It is now
normal cellular prion protein and the infectious form is the shape clear that the prion that caused mad cow disease in England
of the protein, which influences its stability. The normal cellular killed more than 170 people by causing a disease very similar
form, referred to or PrPC (for prion protein, cellular) is readily
destroyed by host cell proteases, as a normal turnover process,
with older molecules being destroyed as new ones are synthe- PrPSC PrPC 1 Both normal
sized. In contrast, infectious prion proteins, referred to as PrPSC (PrPC) and
(for prion protein, scrapie) are less susceptible to degradation abnormal
(PrPSC) proteins
by proteases and become insoluble, leading to aggregation. are present.
Neuron

Brain tissue

2 PrPSC interacts
with PrPC.

3 PrPC is
converted
into PrPSC.

(a)

Spongiform Brain tissue

lesions 4 Conversion
continues
and PrPSC
accumulates.

(b)
FIGURE 13.26 Appearance of a Brain with Spongiform
Encephalopathy (a) Normal brain section. (b) Brain section of
patient with spongiform encephalopathy. FIGURE 13.27 Proposed Mechanism by Which Prions
Propagate
? From these photos, why has this disease been given the name it
has? ? Why would PrPSC accumulate when PrPC does not?
330 Chapter 13 Viruses, Viroids, and Prions

to the Creutzfeldt-Jakob disease. Presumably these people con- decomposing carcasses. The stability of prions makes them diffi-
sumed tissue of infected animals. Thus far, no human deaths cult, if not impossible, to eliminate from the environment.
have been attributed to eating sheep infected with the scrapie Additional information about the role of prions in human disease
agent or deer and elk infected with the prion causing chronic is covered in chapter 26.
wasting disease (CWD).
A good example of how a prion disease spreads in the United MicroAssessment 13.11
States is CWD. Deer at a research facility in Colorado were the Two infectious agents that are structurally simpler than viruses are
first animals to show signs and symptoms of CWD in 1967. viroids and prions. Viroids contain only single-stranded RNA and no
Within 40 years, the disease spread from Colorado to wild deer protein; prions contain protein and no nucleic acid.
populations through their natural migration and transport of cap- 30. Distinguish between a viroid and a prion in terms of structure
tive farmed animals located in more than a dozen states and parts and hosts.
of Canada. The disease is spread through direct animal-to-animal 31. Discuss how prion proteins accumulate in nervous tissue.
(nose-to-nose) contact and as a result of indirect exposure to
32. Must all prion diseases result from eating infected
prions in the environment. Feed, water sources and soil may be food? Explain. +
contaminated with prions shed in saliva, urine and feces, or

FUTURE CHALLENGES 13.1

Gene Therapy: Turning a Corner?
Gene therapy is the treatment of hereditary inside cells in the body to allow synthesis of Has gene therapy achieved any success in
disorders by introducing new genetic informa- the gene products for an extended period of humans? The answer is yes, with encouraging
tion into cells of the body to correct the defect. time. This requires that the immune defenses results. Gene therapy was used to overcome a
The goal is to achieve persistent expression of of the body be circumvented. The genes must fatal inherited disease caused by a deficiency
the introduced normal DNA so as to modify the also be transferred to enough cells in the of an enzyme of purine metabolism that results
defective phenotype without unintended harm- proper tissues, and the genes expressed in in major defects in the body’s immune sys-
ful consequences. Gene therapy was launched large enough amounts, to change the pheno- tem. Ten young children were treated with a
in 1990, and numerous trials treating different type. Not all viruses can transfer genes to non- retrovirus containing the normal gene. All ten
conditions are now underway. However, no dividing cells, so the properties of the target patients are alive after up to 10 years and most
treatments have gained final approval for use tissues must be known. If cells are dividing, are leading normal lives without any adverse
in the United States. It has been a rocky road it may be necessary to use a retrovirus whose effects from the treatment. Gene therapy has
with technical challenges and the death of a DNA becomes part of the host’s genome. If also been used on three individuals suffering
volunteer in one of the trials. this is done, however, the potential exists for from retinal degeneration, which results in
Scientists have focused on viruses as the the DNA to integrate into a vital gene or into total blindness. An adenovirus containing the
most effective way to introduce desired genes genes concerned with the control of growth. normal gene was injected into the retina of
into cells. This is because viral genomes are Thus, another major consideration is how to the eye. The vision of each patient improved
small and readily manipulated, so foreign prevent any unintended consequences result- slightly without any serious adverse effects.
genes can be easily cloned into them. In addi- ing from gene therapy. A brain disorder that usually kills boys before
tion, the attachment proteins of the viral coat Three groups of viruses have been studied they become teenagers has also been treated
can be genetically altered, thereby allowing most extensively as potential vectors. These successfully. The disease results from a defec-
the virion to attach to different tissues. A vari- are the double-stranded DNA adenoviruses, the tive gene involved in maintaining the myelin
ety of well-studied animal viruses are known, single-stranded DNA adeno-associated viruses, sheath around nerves. A normal gene was
some of which integrate their DNA into the and the retroviruses. In all cases, most of the incorporated into blood cells of two 7-year-old
host’s genome, ensuring their maintenance in viral genome has been deleted and replaced boys suffering from the condition. Some of
the cells. with the genes intended to cure the inherited the cells made the functional protein and also
For all its promises, gene therapy has a disease. Such deletions eliminate the pathoge- apparently migrated to the boys’ brains. After
number of problems that must be solved. For nicity of the virus and also reduce its ability to 2 years, the progressive damage has stopped.
one thing, the new genetic information must provoke an immune response that would other-
not only be delivered, but also maintained wise eliminate it.

Summary
13.1 ■ General Characteristics of Viruses Viral Architecture
Most viruses are approximately 100- to 1,000-fold smaller than the cells At a minimum, a virion (viral particle) consists of nucleic acid sur-
they infect (figure 13.1). rounded by a capsid. Capsids are composed of capsomers. Some
 Part II The Microbial World 331

viruses have an envelope surrounding the nucleocapsid; other viruses region of DNA called CRISPR (figure 13.11). That region is transcribed,
are naked (figure  13.2). They contain either RNA or DNA, but never and the spacer segments join with Cas proteins, functioning as a type of
both. The shape of a virus is generally icosahedral, helical, or complex RNA interference to target the phage DNA for destruction.
(figure 13.3).
13.5 ■ Methods Used to Study Bacteriophages
Viral Taxonomy Plaque assays are used to quantitate the phages particles in samples
Viruses are classified primarily on the basis of their genome structure (figure  13.12). Each plaque represents a single phage particle infecting
and hosts they infect (table  13.1). The names of virus families end in a cell.
-viridae. Viruses are also given informal names and are sometimes
grouped on their routes of transmission (table 13.2). 13.6 ■ Animal Virus Replication
The generalized infection cycle of animal viruses can be viewed as a
13.2 ■ Bacteriophages five-step process.

Lytic Phage Infections: T4 Phage as a Model Attachment

Lytic or virulent phages exit the host at the end of the cycle by lysing Attachment proteins or spikes on the virus particle attach to specific
the host, resulting in a productive infection. The infection proceeds receptors on the cell surface.
through five steps: attachment, genome entry, synthesis of phage pro- Penetration and Uncoating
teins and genome, assembly (maturation) and release (figure 13.5).
In the case of animal viruses, the entire virion enters the cell. Enveloped
Temperate Phage Infections: Lambda Phage as a Model viruses either fuse with the host membrane or are taken in by receptor-
Temperate phages have the option of either directing a productive mediated endocytosis (figure  13.13). Naked virions enter by receptor-
infection or initiating a lysogenic infection (figure  13.6); the infected mediated endocytosis. Uncoating releases the nucleic acid from the
cell is a lysogen. A repressor maintains the prophage in an integrated protein coat.
state, but the prophage can be excised to initiate a lytic infection. Synthesis of Viral Proteins and Replication
Consequences of Lysogeny of the Genome (figure 13.14)
Lysogens are immune to superinfection. Lysogenic conversion occurs DNA viruses generally replicate in the nucleus and use the host cell
if a prophage carries genes that change the phenotype of the host cell machinery for DNA synthesis as well as gene expression, although they
(table 13.3). often encode their own DNA polymerase. Replication of ssDNA viruses
is similar to that of double-stranded DNA viruses, but the complemen-
Filamentous Phages: M13 Phage as a Model tary strand must be synthesized first. RNA viruses usually replicate in
Filamentous phages cause productive infections, but the viral particles the cytoplasm. Replication requires a virally encoded replicase to syn-
are continually extruded from the host in the assembly process and the thesize complementary RNA strand. This enzyme lacks proofreading
cells are not killed (figure 13.7). ability and makes more mistakes in replication than DNA polymerase.
Reverse-transcribing viruses encode reverse transcriptase, which
13.3 ■ The Roles of Bacteriophages synthesizes DNA from an RNA template. As with replicases, these
in Horizontal Gene Transfer enzymes are error-prone.
Generalized Transduction Assembly
Generalized transduction results from a phage packaging error during Capsids are formed, and then the genome and any necessary proteins
assembly. Generalized transducing particles can transfer any gene of a are packaged within it. The process may take place in the cytoplasm,
donor cell. nucleus, or in a variety of organelles.
Specialized Transduction Release
Specialized transduction results from an excision mistake made by a Enveloped virions most often exit by budding (figure 13.15). Naked
temperate phage during its transition from a lysogenic to a lytic cycle virions are released when the host cell dies.
(figure 13.9). Only genes located near the site at which the temperate
phage integrates are transduced. 13.7 ■ Categories of Animal Virus Infections (figure 13.16)
Acute infections are characterized by sudden onset of symptoms of
13.4 ■ Bacterial Defenses Against Phages relatively short duration. Persistent infections can continue for the life
of the host, with or without symptoms.
Preventing Phage Attachment
If a bacterium alters or covers a given receptor, that cell becomes resis- Acute Infections
tant to any phage that requires the receptor. On a cellular level, acute infections can be compared to productive lytic
infections by bacteriophages, but even though the cells often die, the
Restriction-Modification Systems host may survive because of the immune response.
Restriction-modification systems protect bacteria from phage infec-
tion by quickly degrading foreign DNA (figure  13.10). Restriction Persistent Infections
enzymes recognize and cut specific DNA sequences; modifi- Chronic infections are characterized by the continuous production of
cation enzymes protect the hosts’ DNA from the action of the restriction low levels of viral particles; latent infections are analogous to lysogeny
enzyme by adding methyl groups to certain nucleobases. in bacteriophages.

CRISPR System 13.8 ■ Viruses and Human Tumors

Bacterial cells that survive some phage infections appear to retain small Some viruses carry oncogenes that interfere with the ability of the cell
segments of phage DNA, termed spacer, and incorporate them into a to control growth. Most virus-induced tumors are caused by certain
332 Chapter 13 Viruses, Viroids, and Prions

DNA viruses (table 13.5). A vaccine against human papillomaviruses 13.10 ■ Plant Viruses
(HPVs) prevents many cervical cancers. Viral infection of plants can be recognized by outward signs such as
yellowing foliage, stunted growth, and tumor formation; plants usually
13.9 ■ Cultivating and Quantitating Animal Viruses do not recover from the infection (figure 13.23). Virions do not bind to
Cultivating Animal Viruses receptor sites on plant cells but enter through wound sites. Insects are
probably the most important transmitter of the viruses.
Cell culture or tissue culture is commonly used to cultivate most
viruses (figure 13.19). This can be done using primary cultures or estab- ■
13.11 Other Infectious Agents: Viroids and Prions
lished cell lines. Many viruses can be detected by their effect on cells
in culture, called a cytopathic effect (figure 13.20). Certain viruses cause Viroids
an infected cell to form an inclusion body. Viroids are plant pathogens that consist of small circular, single-
Quantitating Animal Viruses stranded RNA molecules (figure 13.25).
The plaque assay is one of the most precise methods for determining the Prions
concentration of animal viruses in a sample. In some cases, virions can Prions are composed solely of protein, and cause a number of
be counted with an electron microscope (figure 13.21). Quantal assays transmissible spongiform encephalopathies (table 13.6). Prions accu-
estimate the titer by determining the ID50 or LD50. The concentration of mulate by converting PrPC to PrPSC, proteins that are less susceptible to
viruses able to cause hemagglutination can be measured by determin- proteases and form aggregates (figure 13.27).
ing the highest dilution that clumps red blood cells (figure 13.22).

Review Questions
Short Answer 3. Classification of viruses is based on all of the following except
1. Why are naked viruses generally more resistant to disinfectants a) type of nucleic acid.
than are enveloped viruses? b) shape of virus.
2. How is the replication cycle of lambda phage different from that c) size of virus.
of T4? d) host infected.
3. What is lysogenic conversion? e) strandedness of nucleic acid.
4. How is specialized transduction different from generalized 4. Temperate phages can do all of the following except
transduction? a) lyse their host cells.
5. How does the CRISPR system protect bacteria from phage b) change properties of their hosts.
infection? c) integrate their DNA into the host DNA.
6. Why must (–) strand but not (+) strand RNA viruses bring their d) bud from their host cells.
own replicase into a cell? e) become prophages.
7. Why are RNA viruses and retroviruses more error-prone in their 5. All phages must have the ability to
replication than DNA viruses? 1. have their nucleic acid enter the host cell.
8. What is the role of a prophage in persistent infections? 2. kill the host cell.
9. How do oncogenes differ from proto-oncogenes? 3. multiply in the absence of living bacteria.
10. Describe how prions propagate. 4. lyse the host cell.
5. have their nucleic acid replicate in the host cell.
Multiple Choice
a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5
1. Capsids are composed of 6. Filamentous phages
a) DNA. b) RNA. c) protein. a) infect animal and bacterial cells.
d) lipids. e) polysaccharides. b) cause their host cells to grow more quickly.
2. The tail fibers on phages are associated with c) are extruded from the host cell.
a) attachment. d) undergo assembly in the cytoplasm.
b) penetration. e) degrade the host cells’ DNA.
c) transcription of phage DNA.
d) assembly of virus.
e) lysis of host.
 Part II The Microbial World 333

7. Influenza vaccines must be changed yearly because the amino acid Applications
sequence of the viral proteins change gradually over time. Based 1. A public health physician isolated large numbers of phages from
on this information, which is the most logical conclusion? The rivers used as a source of drinking water in western Africa. The
influenza virus physician is very concerned about humans becoming ill from
a) is enveloped. drinking this water, although she knows that phages specifically
b) is naked. attack bacteria. Why is she concerned?
c) has a DNA genome. 2. Researchers debate the evolutionary value to the virus of its abil-
c) has an RNA genome. ity to cause disease. Many argue that viruses accidentally cause
e) causes a persistent infection. disease and only in animals that are not the natural host. They state
8. Acute infections of animals that this strategy may eventually prove fatal to the virus’s future
in that host. It is reasoned that the animals will eventually develop
1. are a result of productive infection.
immune mechanisms to combat the virus and prevent its spread.
2. generally lead to long-lasting immunity. Another group of researchers supports the view that disease is a
3. result from integration of viral nucleic acid into the host. way to enhance the survival of the virus. What position would you
4. are usually followed by chronic infections. take, and what arguments would you give to support your view?
5. often lead to tumor formation.
a) 1, 2 b) 2, 3 c) 3, 4 d) 4, 5 e) 1, 5 Critical Thinking +
9. Quantitating viral titers of both phage and animal viruses fre- 1. A filter capable of preventing bacteria from passing is placed at
quently involves the bottom of a U tube to separate the two sides. Streptomycin-
a) plaque formation. resistant cells of a bacterial strain are placed on one side of the
b) quantal assays. filter and streptomycin-sensitive cells are placed on the other side.
After incubation, the side of the tube that originally contained only
c) hemagglutination.
streptomycin-sensitive cells now contains some streptomycin-
d) determining the ID50.
resistant cells. Give three possible reasons for this observation.
e) counting of virions by microscopy. What further experiments would you do to determine the correct
10. Prions explanation?
a) contain only nucleic acid without a protein coat. 2. Why is it virtually impossible to eradicate (eliminate) a disease
b) replicate like HIV. caused by a zoonotic virus?
c) integrate their nucleic acid into the host genome.
d) cause diseases of humans.
e) cause diseases of plants.
14 The Innate Immune Response
KEY TERMS
Apoptosis Programmed death
of “self” cells that does not cause
inflammation.
scavenging debris and producing pro-
inflammatory cytokines.
Membrane Attack Complex
(MAC) Complement system
Complement System Series of
proteins in blood and tissue fluids components that assemble to form
that can be activated to help destroy pores in membranes of invading cells.
and remove invading microbes. Neutrophil Major type of
Cytokines Proteins that function as phagocytic cell in blood; neutrophils
chemical messengers, allowing cells quickly move to infected tissues to
to communicate. destroy invading microbes.
Inflammatory Response Opsonization Coating of an object
Coordinated innate response with with molecules for which phagocytes
the purpose of containing a site of have receptors, making it easier for
damage, localizing the response, phagocytosis to occur.
eliminating the invader and Pattern Recognition Receptors
restoring tissue function. The signs (PRRs) Proteins on or in cells that
of inflammation are swelling, heat, recognize specific compounds
redness, and pain in the infected area. unique to microbes or tissue damage,
Innate Immunity Host defenses allowing the cells to sense the
involving anatomical barriers, sensor presence of invading microbes or
systems that recognize patterns damage.
associated with microbes or tissue Phagocyte Cell type that
Human blood cells (color-enhanced scanning electron micrograph). damage, phagocytic cells, and the specializes in engulfing and digesting
inflammatory response. microbes and cell debris.
Macrophage Type of phagocytic Phagocytosis The process by
cell that resides in tissues and which a phagocyte engulfs an
A Glimpse of History has multiple roles, including invader.
As soon as microorganisms were shown to cause disease, scientists
worked to explain how the body defends itself against their invasion.
Elie Metchnikoff, a Russian-born scientist, hypothesized that specialized
cells in the body destroy invading organisms. His ideas arose while he that phagocytic cells were primarily responsible for destroying disease-
was studying the larval form of starfish. As he looked at the larvae under causing organisms. He spent the rest of his life studying this process and
the microscope, he could see ameba-like cells within the bodies. He other biological phenomena. Metchnikoff was awarded a Nobel Prize in
described his observations: 1908 for his studies of immunity.
.  .  . I was observing the activity of the motile cells of a

F 
transparent larva, when a new thought suddenly dawned on me. rom a microorganism’s standpoint, the tissues and fluids of
It occurred to me that similar cells must function to protect the the human body are much like a culture flask filled with a
organism against harmful intruders. . . . I thought that if my warm nutrient-rich solution. Considering this, it may be sur-
guess was correct a splinter introduced into the larva of a star- prising that the interior of the body—including blood, muscles,
fish should soon be surrounded by motile cells much as can be
bones, and organs—is generally sterile. If this were not the case,
observed in a man with a splinter in his finger. No sooner said
microbes would simply degrade our tissues, just as they readily
than done. In the small garden of our home .  .  . I took several
rose thorns that I immediately introduced under the skin of some break down the bodies of dead animals.
beautiful starfish larvae which were as transparent as water. Very How do body tissues remain sterile in this world full of microbes?
nervous, I did not sleep during the night, as I was waiting for the Like other multicellular organisms, humans have several mechanisms
results of my experiment. The next morning, very early, I found of defense. First, we are covered with skin and mucous membranes
with joy that it had been successful. that prevent most microbes from entering the tissues. If microbes
get past those barriers, sensor systems in the tissues detect them and
Metchnikoff reasoned that certain cells in animals are able to ingest
and destroy foreign material. He called these cells phagocytes (“cells that then direct and assist other defenses that destroy the invaders. Innate
eat”) and proposed they were primarily responsible for the body’s ability immunity is the routine protection provided by these mechanisms.
to destroy invading microbes. He then studied the process by watching Innate immunity had been called a non-specific defense,
phagocytes ingest and destroy invading yeast cells in transparent water but recent discoveries show that most of its components detect
fleas. In 1884, Metchnikoff published a paper supporting his belief molecules associated with invading microbes or tissue damage,
334
 Part III Microorganisms and Humans 335
Focus Figure

First-line defenses Skin and mucous membranes

Prevent microbial entry

Microbial invasion

Sensor systems Pattern recognition receptors Pattern recognition receptors Complement system
Detect microbial (surfaces, endosomes, and (cytoplasm of many cell types) (blood and tissue fluids)
invasion phagosomes of sentinel cells)

Innate effector actions Inflammatory Inflammatory Interferon Inflammatory response

Destroy invader response response response Opsonization
Membrane attack complexes

FIGURE 14.1 Overview of the Innate Defenses ? What is the role of the sensor systems in innate immunity?

a feature called pattern recognition. The molecules recognized First-line defenses are the barriers that separate and shield the
include parts of bacterial cell walls and other compounds unique interior of the body from the surrounding environment; they are
to microbes, as well as substances associated with damaged host the initial obstacles microbes must overcome to invade the tissues.
cells. bacterial cell walls, p. 58 The anatomical barriers, which include the skin and mucous mem-
In addition to the innate response, vertebrates have evolved branes, not only provide physical separation but are often bathed
a more specialized defense system, providing protection called in secretions that have antimicrobial properties.
adaptive immunity. This develops throughout life and substan- Sensor systems within the body recognize when the first-line bar-
tially increases the host’s ability to defend itself. Each time the riers have been breached and then relay that information to other com-
body is exposed to a microbe, or certain other types of foreign ponents of the host defenses. Certain cells serve as sentinels (lookouts
material, the adaptive defense system first “learns” and then or guards). They have pattern recognition receptors (PRRs) on their
“remembers” the most effective response to that specific material; surface and within their endosomes or phagosomes. These receptors
it then reacts accordingly if the material is encountered again. The recognize groups of compounds unique to microbes, allowing the sen-
substance that causes an immune response is called an antigen. tinel cells to detect invaders. When invasion is detected, the cells send
An important action of the adaptive immune response is the chemical signals to alert other components of the host defenses, trig-
production of Y-shaped proteins called antibodies. These bind gering a protective response (an effector action). Many cell types have
specifically to antigens, thereby targeting them for destruction or a different set of PRRs in their cytoplasm, allowing them to recognize
removal by other host defenses. The adaptive immune response when they have been invaded by a microbe. As with the sentinel cells,
can also destroy the body’s own cells—referred to as host cells or they respond by sending out a call for help. Another type of sensor
“self ” cells—if they harbor a virus or other invader. is a series of proteins always present in blood and tissue fluids; these
To simplify the description of the immune system, it is help- proteins are collectively called the complement system because they
ful to consider it as a series of individual parts. This chapter will can “complement” (act in combination with) the adaptive immune
focus almost exclusively on innate immunity. Remember, how- defenses. The complement system becomes activated in response to
ever, that although the various parts are discussed separately, their certain stimuli, setting off a chain of events that results in removal and
actions are connected and coordinated. In fact, as you will see in destruction of invading microbes. endosome, p. 72 phagosome, p. 72
chapter 15, certain components of the innate defenses educate the When invading microorganisms or tissue damage is detected,
adaptive defenses, helping them recognize that a particular antigen an inflammatory response occurs, involving many components
represents a microbial invader. of the innate defenses. During this response, cells that line local
blood vessels undergo changes that allow complement system
components and other proteins to leak out into tissues. Phagocytes
14.1 ■ Overview of the (cells that specialize in engulfing and digesting microbes and cell
Innate Defenses debris) also leave the bloodstream and accumulate in the tissues.
Some types of phagocytes play a dual role, destroying invaders
Learning Outcome while communicating with cells of the adaptive immune system,
1. Outline the essential components of the innate defenses. enlisting their far more powerful effects. Figure 14.1 provides a
general overview of innate immunity.
336 Chapter 14 The Innate Immune Response

MicroAssessment 14.1 Physical Barriers

First-line defenses are the initial barriers that microbes must pass to All exposed surfaces of the body are lined with epithelial cells
invade tissues. Sensor systems within the body recognize invading (figure 14.3). These cells are tightly packed together and rest on a
microbes. Inflammation is a coordinated response to invasion or thin layer of fibrous material, the basement membrane.
tissue damage that recruits phagocytes and other protective
mechanisms to an area.
Skin
1. List two sensor systems of the innate defenses. The skin, an obvious visible barrier, is the most difficult for
2. Describe the dual roles played by some types of phagocytes. microbes to penetrate. It is composed of two main layers—the
3. What molecules might pattern recognition receptors dermis and the epidermis (see figure 22.1). The dermis contains
recognize? + tightly woven fibrous connective tissue, making it extremely
tough and durable (the dermis of cattle is used to make leather).
The epidermis is composed of many layers of epithelial cells
14.2 ■ First-Line Defenses that become progressively flattened toward the exterior. The
outermost sheets are made up of dead cells filled with a water-
Learning Outcome
repelling protein called keratin, resulting in the skin being a dry
2. Describe the first-line defenses, including the physical barriers,
antimicrobial substances, and normal microbiota.

The body’s borders serve as the first line of defense against

invading microbes (figure  14.2). Some of these borders are
thought of as being “inside” the body, but they directly con-
tact the external environment. For example, the digestive tract,
which begins at the mouth and ends at the anus, is simply a hol-
low tube that runs through the body, allowing intestinal cells to
absorb nutrients from food that passes through (see figure 24.1); Nucleus
the respiratory tract is a cavity that allows O2 and CO2 to be
exchanged (see figure 21.1).
In this section, we will describe the general physical and
Basement
chemical aspects of the anatomical barriers, as well as the protec- membrane
tive contributions of the normal microbiota. These are described in
Connective
more detail in the chapters dealing with each body system. tissue

Mouth
Eye
Stratified epithelium
Respiratory tract • Skin (the outer cell layers are embedded with keratin)
• Lining of the mouth, vagina, urethra, and anus

Cilia
Digestive tract
Mucus-
producing cell
Columnar cell
Urogenital tract

Skin
Anus Skin
Columnar epithelium
Mucous • Passages of respiratory system
membranes • Various tubes of the reproductive systems
FIGURE 14.2 The Body’s Borders These borders separate the
interior of the body from the surrounding environment; they are the
initial obstacles microorganisms must overcome to invade tissues. FIGURE 14.3 Epithelial Barriers Cells of these barriers are tightly
The skin is shown in tan, and mucous membranes in pink. packed together and rest on a layer of thin fibrous material, the
basement membrane.
? Why are the contents of the digestive tract considered to be
outside the body? ? What is the purpose of the cilia on the respiratory epithelium?
 Part III Microorganisms and Humans 337

environment. The cells continually slough off, taking with them Antimicrobial factors
any microbes that might be adhering.   anatomy, physiology, and in saliva (lysozyme,
peroxidase, lactoferrin)
ecology of the skin, p. 522

Lysozyme in tears
Mucous Membranes and other secretions
and in phagocytes
Mucous membranes line the digestive tract, respiratory tract, Normal microbiota
and genitourinary tract. They are constantly bathed with mucus Mucus, cilia
Removal of inhaled
or other secretions that help wash microbes from the surface. particles
Most mucous membranes have mechanisms that move microbes
toward areas where they can be eliminated. For example,
Physical barrier
peristalsis—the contractions of the intestinal tract—propels of skin, salty
food and liquid and also helps remove microbes. The respira- residue, fatty acids,
tory tract is lined with ciliated cells; the hairlike cilia con- normal microbiota
stantly beat in an upward motion, moving materials away from
the lungs to the throat where they can then be swallowed. This Acid in stomach
(low pH)
movement out of the respiratory tract is referred to as the
mucociliary escalator. cilia, p. 74
Rapid pH change Normal
from stomach to microbiota
upper intestine
Antimicrobial Substances
Skin and mucous membranes are protected by a variety of
Flushing of
antimicrobial substances that inhibit or kill microorganisms urinary tract
pH and normal
(figure  14.4). For example, the salty residue that accumulates microbiota of vagina
on skin as perspiration evaporates inhibits all but salt-tolerant
microbes. FIGURE 14.4 Antimicrobial Substances and the Normal
Lysozyme, the enzyme that degrades peptidoglycan, is Microbiota These play important roles in protecting the body’s
in tears, saliva, and mucus. It is also found within the body, borders.
in phagocytic cells, blood, and the fluid that bathes tissues. ? How is lysozyme antibacterial?
lysozyme, p. 62
Peroxidase enzymes, which break down hydrogen perox-
ide to produce reactive oxygen species, are in saliva and milk;
they are also found within body tissues and inside phagocytes. prevents pathogens from adhering to host cells by covering binding
Bacteria that produce the enzyme catalase can break down sites that might otherwise be used for attachment. The population
hydrogen peroxide, potentially destroying it before peroxidase also consumes available nutrients that could otherwise support the
reacts with it. Catalase-negative organisms are more readily growth of less desirable organisms.
killed by peroxidase. reactive oxygen species, p. 90 catalase, p. 90 Some members of the normal microbiota produce com-
Lactoferrin is an iron-binding protein in saliva, mucus, pounds toxic to other bacteria. In the hair follicles of the
and milk; it is also found in some types of phagocytes. A simi- skin, for instance, Propionibacterium species degrade lipids,
lar compound, transferrin, is in blood and tissue fluids. Iron releasing fatty acids that inhibit the growth of many patho-
is one of the major elements, so withholding it prevents micro- gens. In the gastrointestinal tract, some strains of E. coli syn-
bial growth. Some microorganisms can capture iron from the thesize colicins, a group of proteins toxic to certain bacteria.
host, however, thwarting this defense. major elements, p. 92 Lactobacillus species growing in the vagina produce lactic
Defensins are short antimicrobial peptides produced by neu- acid as a fermentation end product, resulting in an acidic pH
trophils and epithelial cells. They insert into bacterial membranes, that inhibits the growth of some pathogens.
forming pores that damage cells. Disruption of the normal microbiota, which occurs when
antibiotics are used, can predispose a person to various infections.
Examples include antibiotic-associated diarrhea and pseudomem-
Normal Microbiota (Flora) branous colitis, caused by the growth of toxin-producing strains of
The normal microbiota (flora) is the population of micro- Clostridium difficile in the intestine, and vulvovaginitis, caused by
organisms that routinely grow on the body surfaces of healthy excessive growth of Candida albicans in the vagina. Clostridium
humans (see figure  16.1). Although these organisms are not difficile–associated disease, p. 594 vulvovaginal candidiasis, p. 618
technically part of the immune system, they provide consider- The normal microbiota is also essential to the development
able protection. of the immune system. As certain microbes are encountered, the
One protective effect of the normal microbiota is the competi- system learns to distinguish harmless ones from pathogens. Other
tive exclusion of pathogens. For example, the normal microbiota effects of the population will be discussed in chapter 16.
338 Chapter 14 The Innate Immune Response

MicroAssessment 14.2 The cells of the immune system can move from one part of
the body to another, traveling through the body’s circulatory
Physical barriers that prevent microbes from entering the body systems like vehicles on an extensive interstate highway sys-
include skin and mucous membranes. Antimicrobial substances,
including lysozyme, peroxidase enzymes, lactoferrin, and defensins,
tem. They are always found in normal blood, but their numbers
are on body surfaces. The normal microbiota excludes pathogens and usually increase during infections, recruited from reserves of
promotes immune system development. immature cells in the bone marrow. Some of the cell types are
4. How does the mucociliary escalator protect against infection? found primarily in the blood, but others leave the blood circula-
tory system and take up residence in various tissues. Certain cell
5. What is the role of lactoferrin?
types play dual functions, having crucial roles in both innate and
6. How would damage to the ciliated cells of the respiratory tract
adaptive immunity.
predispose a person to infection? +
The formation and development of blood cells is called
hematopoiesis (Greek for “blood” and “to make”). All blood
14.3 ■ The Cells of the cells, including those important in the body’s defenses, originate
Immune System from the same cell type, the hematopoietic stem cell, found
in the bone marrow (figure  14.5). As with other types of stem
Learning Outcome cells, hematopoietic cells are capable of long-term self-renewal,
meaning they can divide repeatedly. Hematopoietic stem cells are
3. Describe the characteristics and roles of granulocytes,
mononuclear phagocytes, dendritic cells, and lymphocytes. induced to develop into the various types of blood cells by a group
of proteins called colony-stimulating factors (CSFs).

FIGURE 14.5 Blood Cells and Their Derivatives All of Hematopoietic stem cell
these descend from hematopoietic stem cells found in the bone (in bone marrow)
marrow. Multiple steps occur between the hematopoietic stem cell
and the final cells produced.
? How are the roles of neutrophils
and macrophages similar? Self-
renewal

Common Common
myeloid progenitor lymphoid progenitor

Erythroblast Megakaryoblast Putative mast Myeloblast Monoblast Lymphoblasts

cell precursor

Megakaryocyte

Eosinophil Basophil Neutrophil Monocyte Natural T cell B cell

killer (NK) cell
Red blood cell Platelets Granulocytes Lymphocytes
(erythrocyte) (thrombocytes)

Mast cell Macrophage Dendritic cell

White blood cells (leukocytes)

 Part III Microorganisms and Humans 339

The general categories of blood cells and their derivatives Neutrophils efficiently engulf and destroy bacteria and
include red blood cells, platelets, and white blood cells. Red blood other material. Their granules, which stain poorly, contain
cells, or erythrocytes, carry O2 in the blood. Platelets, which are many enzymes and antimicrobial substances that help destroy
actually fragments arising from large cells called megakaryocytes, the engulfed materials. Neutrophils are the most numerous and
are important for blood clotting. White blood cells, or leukocytes, important granulocytes of the innate responses. They are also
are important in all host defenses. Leukocytes can be divided into called polymorphonuclear neutrophilic leukocytes, polys, or
four broad groups—granulocytes, mononuclear phagocytes, den- PMNs, names that reflect the appearance of multiple lobes of
dritic cells, and lymphocytes (table 14.1). their single nucleus. They normally account for over half of the
circulating white blood cells, and their numbers increase during
most bacterial infections. Few neutrophils are generally found in
Granulocytes tissues, except during inflammation. Because of their importance
Granulocytes contain cytoplasmic granules filled with biologi- in innate immunity, they will be described in more detail later in
cally active chemicals. There are three types of granulocytes— the chapter.  specialized attributes of neutrophils, p. 348
neutrophils, basophils, and eosinophils; their names reflect the Basophils are involved in allergic reactions and inflam-
staining properties of their granules. mation. Their granules, which stain dark purplish-blue with

TABLE 14.1 Leukocytes

Cell Type (% of blood leukocytes) Location in Body Function

Granulocytes
Neutrophils (polymorphonuclear Most common type of Phagocytize and digest engulfed
neutrophilic leukocytes or PMNs, circulating leukocyte; few materials
often called polys; 55–65%) in tissues except during
inflammation
Eosinophils (2–4%) Few in tissues except in certain Participate in inflammatory
types of inflammation and reaction and immunity to some
allergies parasites

Basophils (0–1%), mast cells Basophils in circulation; mast Release histamine and other
cells present in most tissues inflammation-inducing chemicals
from the granules

Mononuclear Phagocytes
Monocytes (3–8%) In circulation; they Phagocytize and digest engulfed
differentiate into either materials
macrophages or dendritic cells
when they migrate into tissue

Macrophages Present in virtually all tissues; Phagocytize and digest engulfed

known by various names based materials
on the tissue in which they are
found

Dendritic cells Initially in tissues, but they Collect antigen from the tissues
migrate to secondary lymphoid and then bring it to lymphocytes
organs (e.g., lymph nodes, that gather in the secondary
spleen, appendix, tonsils) lymphoid organs

Lymphocytes
Several types (25–35%) In lymphoid organs (e.g., Participate in adaptive immune
lymph nodes, spleen, thymus, responses
appendix, tonsils); also in
circulation
340 Chapter 14 The Innate Immune Response

the basic dye methylene blue, contain histamine and other

chemicals that increase capillary permeability during inflam- Microglial cells
mation. Mast cells are similar in appearance and function to in the brain
basophils but are found in tissues rather than blood. They do
not come from the same precursor cells as basophils. Mast
cells are important in the inflammatory response and are
Resident and
responsible for many allergic reactions. recirculating
Eosinophils are thought to be primarily important in rid- Alveolar macrophages
ding the body of parasitic worms. They are involved in aller- macrophages in the lymph
in the lungs nodes
gic reactions, causing some of the symptoms associated with
allergies, but reducing others. The granules of eosinophils, Macrophages
and blood
which stain red with the acidic dye eosin, contain antimicro- Kupffer cells in monocytes
bial substances and also histaminase, an enzyme that breaks the liver in the spleen
down histamine.
Mesangial
phagocytes
in the kidneys
Mononuclear Phagocytes
Mononuclear phagocytes make up the mononuclear phago-
cyte system (MPS) (figure  14.6). This grouping includes
monocytes—which circulate in the blood—and the cell types
that develop from them as they leave the bloodstream and
migrate into tissues.
Macrophages are a differentiated form of monocytes, Precursors in
Peritoneal macrophages
meaning they have gained specialized properties. They are in the abdominal cavity bone marrow
an important sentinel cell, present in nearly all tissues, and
particularly abundant in the liver, spleen, lymph nodes, lungs,
and the peritoneal (abdominal) cavity. When residing in tis-
sues, they are given different names based on their location
(figure  14.6). Macrophages will be discussed in more detail
later in the chapter.  specialized attributes of macrophages, p. 348
Dendritic cells also develop from monocytes. Their function
goes beyond engulfment and destruction of an invader, so they
will be considered separately.

Dendritic Cells
Dendritic cells are sentinel cells that function as “scouts.”
They engulf material in the tissues and then bring it to the cells FIGURE 14.6 Mononuclear Phagocyte System Cells in this
of the adaptive immune system for “inspection.” Most den- system sometimes have special names depending on their location—
dritic cells develop from monocytes, but some descend from for example, Kupffer cells (in the liver) and alveolar macrophages
other cell types. Details regarding the interactions of dendritic (in the lung).
cells with the cells of the adaptive immune response will be ? Macrophages descend from which type of blood cell?
discussed in chapter 15.

Lymphocytes MicroAssessment 14.3

Lymphocytes are responsible for adaptive immunity, the focus Granulocytes include neutrophils, basophils, and eosinophils.
of chapter 15. In contrast to the generic pattern recognition that Mononuclear phagocytes include monocytes and macrophages.
characterizes the innate defenses, cells of the two major groups of Dendritic cells function as scouts for the adaptive immune system.
lymphocytes, B cells and T cells, are remarkably specific in their Lymphocytes are responsible for adaptive immunity.
recognition of antigen. These cell types generally reside in lymph 7. Which type of granulocyte is the most abundant?
nodes and other lymphatic tissues. Natural killer (NK) cells are 8. How are macrophages related to monocytes?
another type of lymphocyte; unlike B cells and T cells, however,
9. Why can bone marrow transplants be used to replace defective
they lack specificity in their mechanisms of antigen recognition. lymphocytes? +
lymphatic tissues, p. 357
 Part III Microorganisms and Humans 341

14.4 ■ Cell Communication act together or in sequence, in a complex fashion. The source and
effects of some cytokines are listed in table 14.2.
Learning Outcome Chemokines are cytokines important in chemotaxis of
4. Describe the characteristics and roles of surface receptors,
immune cells. Certain types of cells have receptors for chemo-
cytokines, and adhesion molecules in innate immunity. kines, allowing the cells to sense the location where they are
needed, such as an area of inflammation.
In order for immune system cells to mount a coordinated response Colony-stimulating factors (CSFs) are important in the
to invasion, they must communicate with each other. They do this multiplication and differentiation of leukocytes (see figure 14.5).
through surface receptors, cytokines, and adhesion molecules. When more leukocytes are needed during an immune response,
a variety of different colony-stimulating factors direct immature
cells into the appropriate maturation pathways.
Surface Receptors Interferons (IFNs) are important in the control of viral infec-
Surface receptors can be viewed as the “eyes” and “ears” of a tions. In addition to being antiviral, IFN-gamma helps regulate
cell. They are proteins that generally span the plasma membrane, the function of cells involved in the inflammatory response and
connecting the outside of the cell with the inside, allowing the adjusts certain actions of adaptive immunity. The role of interfer-
inner workings of the cell to sense and respond to external ons during viral infections will be described later in the chapter.
signals. Each receptor is specific with respect to the compound Interleukins (ILs) are produced by leukocytes and have
or compounds it will bind; a molecule that can bind to a given diverse, often overlapping, functions. As a group, they are impor-
receptor is called a ligand for that receptor. When a ligand tant in both innate and adaptive immunity.
binds to its surface receptor, the internal portion of the receptor Tumor necrosis factor (TNF) was discovered because of
becomes modified in some manner. This change then triggers its role in killing tumor cells, a characteristic reflected by the
some type of response by the cell, such as chemotaxis. Cells name, but it has multiple roles. It helps initiate the inflammatory
can alter the types and numbers of surface molecules they make, response and triggers the process of “cell suicide,” a programmed
allowing them to respond to signals relevant to their immediate cell death called apoptosis.  apoptosis, p. 350
situation. chemotaxis, p. 64 Groups of cytokines often act together to generate a response.
For example, certain cytokines referred to as pro-inflammatory
cytokines (TNF, IL-1, IL-6, and others) contribute to inflamma-
Cytokines tion. Others are involved in promoting antibody responses (IL-4
Cytokines can be viewed as the “voices” of a cell. A cytokine and others). A different group stimulates certain types of T cells
produced by one cell diffuses to another and binds to the appro- (IL-2, IFN-gamma, and others).
priate cytokine receptor of that cell. Binding of a cytokine to its
MicroByte
receptor induces a change in the cell such as growth, differentia-
HIV takes advantage of two chemokine receptors, CCR5 and
tion, movement, or cell death. Cytokines act at extremely low con- CXCR4, using them as attachment sites for infection.
centrations, having local, regional, or systemic effects. They often

TABLE 14.2 Some Important Cytokines

Cytokine Source Effect

Chemokines Various cells Chemotaxis

Colony-Stimulating Factors (CSFs) Various cells Stimulate growth and differentiation of different kinds of leukocytes
Interferons
Interferon alpha Leukocytes Antiviral
Interferon beta Fibroblasts Antiviral
Interferon gamma T cells, NK cells Macrophage activation; promotes certain adaptive immune
responses
Interleukins (ILs)
IL-1 Macrophages, epithelial cells T-cell activation; macrophage activation; induces fever
IL-2 T cells T-cell proliferation
IL-4 T cells, mast cells Promotes antibody responses
IL-6 T lymphocytes, macrophages T- and B-cell growth; inflammatory response; fever
Tumor Necrosis Factor (TNF) Macrophages, T cells, Promotes inflammation; cytotoxic for some tumor cells; regulates
NK cells certain immune functions
342 Chapter 14 The Innate Immune Response

Adhesion Molecules (figure  14.7). Others are in phagosomal or endosomal mem-

branes, facing the lumen of the organelle. These let macrophages
Adhesion molecules on the surface of cells allow those cells to
and dendritic cells characterize the material they ingested.
“grab” other cells. For example, when phagocytic cells in the
endosome, p. 72 phagosome, p. 72 lumen, p. 69
blood are needed in tissues, the endothelial cells that line the blood
When a TLR detects a compound, a signal is transmitted
vessels synthesize adhesion molecules that bind to passing phago-
to the cell’s nucleus, causing certain genes to be expressed.
cytic cells. This slows the rapidly moving phagocytes, allowing
The actual response is dictated by the cell type and the array
them to then leave the bloodstream. Other types of adhesion
of TLRs triggered, so it can be tailored to the situation and
molecules are used by cells to form connections so that one cell
category of pathogen. For example, when macrophages “see”
can deliver cytokines or other molecules directly to another cell.
bacterial products, they begin producing pro-inflammatory
cytokines, leading to an inflammatory response. If a TLR
MicroAssessment 14.4 detects viral nucleic acid, the cell synthesizes products that
Surface receptors allow a cell to detect molecules present outside of promote an antiviral response.
that cell. Cytokines provide cells with a mechanism of communication.
Adhesion molecules allow one cell to adhere to another.
10. What is a ligand? NOD-Like Receptors (NLRs)
11. What is the function of a colony-stimulating factor? NOD-like receptors (NLRs) are cytoplasmic proteins that detect
12. How could colony-stimulating factors be used as a therapy? + bacterial components, allowing the cell to recognize when its own
borders have been breached; some also detect signs of cell damage
(figure 14.8). At least 23 NLRs have been described, but details
14.5 ■ Pattern Recognition about their roles are still being uncovered.
When NLRs detect PAMPs or DAMPs, they unleash a series
Receptors (PRRs) of events that lead to outcomes that protect the host, sometimes at
the expense of the cell itself. In macrophages, some NRLs can join
Learning Outcome with other proteins in the cytoplasm to form an inflammasome.
5. Describe the role of TLRs, NLRs, RLRs, and the interferon
response in the host defenses.

Pattern recognition receptors (PRRs) allow the body’s cells Detects

Phagosome Detects lipopolysaccharide Detects
to “see” signs of microbial invasion. They detect generic or endosome peptidoglycan (LPS) flagellin
microbe-associated patterns, called pathogen-associated
molecular patterns (PAMPs). These substances are common
on all microbes, not just pathogens, so they are sometimes Outside
referred to as microbe-associated molecular patterns of cell
(MAMPs). You’ll recognize many of the PAMPs
from earlier reading in the textbook—various cell
wall components (lipopolysaccharide, peptidogly-
can, lipoteichoic acid, and lipoproteins), flagellin
subunits, and RNA  molecules that characterize
TLRs in cytoplasmic membrane
viruses. Some PRRs recognize danger-associated
Cytoplasm
molecular patterns (DAMPs), molecules that indicate
TLRs in phagosomal or endosomal membrane
host cell damage. Although PRRs were discovered relatively
recently, and much is still being learned about them, they have
stimulated tremendous interest in innate immunity.

Toll-Like Receptors (TLRs)

Toll-like receptors (TLRs) are anchored in membranes of senti-
nel cells such as macrophages, dendritic cells, and cells that line
Lumen of
sterile body sites. A number of different TLRs have been described endosome
(at least 10 in humans), and each recognizes a distinct compound Detects Detects Detects
or group of compounds associated with microbes. TLRs are dsRNA bacterial DNA ssRNA
related to Toll receptors, first identified in Drosophila species
FIGURE 14.7 Toll-Like Receptors (TLRs) These pattern
(fruit flies). Their name reflects the reaction of a researcher to the recognition receptors are anchored in membranes of sentinel cells,
sight of a fruit-fly with a mutant form of the gene—she exclaimed allowing these cells to “see” microbial compounds that originated
“toll!”— a German word meaning “awesome.” outside of the cell. Not all of the compounds recognized by TLRs are
Some TLRs are anchored in plasma membranes, allow- shown in this figure.
ing cells to “see” PAMPs in the extracellular environment ? Why would cells that line the blood vessels have TLRs?
 Part III Microorganisms and Humans 343

NLR - Detects flagellin RLR - Detects dsRNA This complex then activates a potent pro-inflammatory cytokine,
thereby initiating an inflammatory response. Mutations in the
genes encoding NLRs seem to be a predisposing factor in certain
inflammatory diseases such as Crohn’s disease (an inflammatory
bowel disease). Scientists hope that by studying NLRs, they will
better understand a wide range of inflammatory disorders.

RIG-Like Receptors (RLRs)

RIG-like receptors (RLRs) are cytoplasmic proteins that
detect viral RNA (figure 14.8). As with pattern recognition
by NLRs, this provides a mechanism for an infected cell to
detect the invader. RLRs can distinguish viral RNA from
NLR - Detects normal cellular RNA because at least two characteristics
peptidoglycan RLR - Detects differ. First, viral  RNA often has three phosphates at the 5'
uncapped ssRNA
NLR - Detects compounds end; recall that cellular RNA is processed after transcription,
that indicate cell damage and one of those steps (capping) modifies the 5' end, hiding
the phosphates. Second, viral RNA is often double-stranded.
FIGURE 14.8 NOD-Like Receptors (NLRs) and RIG-Like
Receptors (RLRs) These pattern recognition receptors are found RNA viruses other than retroviruses routinely generate long
within cells and detect microbial components that indicate cell dsRNA as a result of viral replication. Even DNA viruses often
invasion. Not all NLRs and RLRs are shown. generate long dsRNA because both strands are sometimes
? How do NLRs and RLRs differ from TLRs with respect to the used as templates for transcription, leading to production of
source of the microbial compounds detected? complementary RNA molecules. Cellular RNA is typically not
double-stranded because only one
DNA strand in a gene is used as
Virus ssRNA dsRNA a template for mRNA synthesis.
capping, p. 176

The Interferon Response

Long dsRNA When a cell’s RLRs detect viral
activates genes RNA, the cell responds by syn-
for IFN synthesis.
thesizing and secreting interferons
IFN
(figure  14.9). These proteins then
IFN
attach to specific receptors on both
IFN
the infected cell and neighboring
Cell 1: Cell is infected by a Cell 1: Productive cells, causing the cells to express
IFN diffuses to Virus infects
virus; viral replication infection; cell is destroyed what can be viewed as inactive
neighboring neighboring
produces long dsRNA, as a result of infection.
which induces the cells. cells. “suicide enzymes” (protein kinase
synthesis and secretion of R, RNase L, and others). For conve-
interferon (IFN). nience, we will refer to these collec-
tively as inactive antiviral proteins
(iAVPs). The activated forms of the
IFN
antiviral proteins (AVPs) degrade
mRNA and stop protein synthesis,
IFN activates
leading to apoptosis (a programmed
iAVP genes for iAVP cell death). A key feature of this
synthesis. iAVP AVP

Induction of
iAVP programmed
cell death
(apoptosis)

Cell 2: Interferon induces synthesis of a Cell 2: Infection of cell (detected by the

group of inactive antiviral proteins (iAVP). presence of long dsRNA) results in activation FIGURE 14.9 Antiviral
These have no effect on the cell unless they of the antiviral proteins, leading to apoptosis. Effects of Interferon
are activated. Although the cell dies, the virus does not
have the opportunity to replicate, thus ? Why would it be beneficial to a
preventing viral spread. host for a virally infected cell to
undergo apoptosis?
344 Chapter 14 The Innate Immune Response

response is that the iAVPs are activated by dsRNA. Thus, when 14.6 ■ The Complement System
cells bind interferon, only the infected ones are sacrificed. Their
uninfected counterparts remain functional but are prepared to Learning Outcome
undergo apoptosis should they become infected.  apoptosis, p. 350 6. Describe the three pathways that lead to complement system
activation and the three outcomes of activation.
MicroAssessment 14.5
The complement system is a series of proteins that circulate in
Cells use TLRs to detect microbial compounds that originated outside
of the cell. NLRs and RLRs detect microbial compounds within an the blood and the fluid that bathes the tissues (figure 14.10). Their
infected cell. Viral RNA triggers an interferon response. name is derived from the observation that they “complement” the
function of antibodies. Each of the major complement system pro-
13. Why would macrophages have TLRs in their endosomal
membranes? teins has been given a number along with the letter C (for comple-
ment). The nine major proteins, C1 through C9, were numbered in
14. If a cell produces antiviral proteins, what happens to that cell
when those proteins encounter long dsRNA? the order of their discovery and not the order in which they react.
When a complement protein is split into two fragments, those
15. Why would the discovery of TLRs alter the view that
innate immunity is non-specific? + fragments are distinguished by adding a lowercase letter to each
name. For example, C3 is split into C3a and C3b.

Alternative pathway Lectin pathway Classical pathway

Triggered by Triggered by Triggered by

C3b binding to microbial invaders Mannose-binding lectin (MBL) binding to Antibodies binding to microbial invaders
microbial invaders
Antibody
C3b MBL

Formation of C3 convertase

C3 Splits C3 Opsonization
C3b binds to microbial cells,
Inflammatory response functioning as an opsonin.
C3a and C5a induce changes that
contribute to local vascular permeability
and attract phagocytes. C3a C3b
C3b

C5 Combines with C3
convertase to form an
enzyme that splits C5

C5a C5b

Lysis of foreign cells C5b

C5b combines with complement C6
FIGURE 14.10 Complement proteins C6, C7, C8, and C9 C7 C9
System The three pathways of complement to form membrane attack C9 C9 C9
system activation converge, leading to the same complexes that insert C8
three outcomes (the inflammatory response, lysis into cell membranes.
of foreign cells, opsonization). Not all of the
steps in these pathways are shown.
? How can C3b be both a product of
complement activation and an activator of
the complement system?
 Part III Microorganisms and Humans 345

Complement proteins routinely circulate in an inactive form, cells or other foreign particles. This has two effects: (1) con-
but in response to certain signals indicating the presence of micro- tinued complement activation via the alternative pathway, and
bial invaders, a cascade of reactions occurs. This results in the (2) opsonization. Material that has been opsonized (meaning
rapid activation of the system. The activated forms of the comple- “prepared for eating”) is easier for phagocytes to bind to and
ment proteins have specialized functions that cooperate with other engulf because phagocytes have receptors that attach specifically
host defenses to quickly remove and destroy the invader. to the opsonins (in this case, C3b).

MicroByte
Complement System Activation Think of opsonization as coating a microbe with one layer of Velcro,
with phagocytes having the opposing layer on their surface.
The complement system can be activated by three different
pathways that converge when a complex called C3 convertase is
formed (figure 14.10). C3 convertase then splits C3, leading to
additional steps of the activation cascade. Inflammatory Response
The complement component C5a is a potent chemoattractant,
Alternative Pathway drawing phagocytes to the area where the complement system has
The name of the alternative pathway may seem to imply that been activated. In addition, C3a and C5a induce changes in the
the pathway is “second choice,” but it actually reflects the fact endothelial cells that line the blood vessels, contributing to the
that the pathway was not discovered first. It is quickly and easily vascular permeability associated with inflammation. They also
triggered, providing vital early warning that an invader is present. cause mast cells to release various pro-inflammatory cytokines.
The alternative pathway is triggered when C3b binds to for-
eign cell surfaces. The binding of C3b allows other complement
proteins to then attach, eventually forming the C3 convertase. Lysis of Foreign Cells
What might seem confusing is the fact that C3b is a product of Complexes of complement system proteins (C5b, C6, C7,
complement activation, yet it also triggers the alternative path- C8, and multiple C9 molecules) spontaneously assemble in
way. How can it be both a product and a trigger? This can occur cell membranes, forming doughnut-shaped structures called
because C3 is somewhat unstable, and spontaneously splits to membrane attack complexes (MACs) (figure  14.11). This
C3a and C3b at a low rate even when the complement system has creates pores in the membrane, disrupting the integrity of the
not been activated. The C3a and C3b formed this way are rapidly cell. MACs have little effect on Gram-positive bacteria because
inactivated by regulatory proteins, but some C3b is always present the peptidoglycan layer of these cells prevents the complement
to trigger the alternative pathway when needed. system components from reaching their cytoplasmic membranes.
The outer membranes of Gram-negative bacteria, however, make
Lectin Pathway them susceptible.
Activation of the complement system via the lectin pathway
involves pattern recognition molecules called mannose-binding
lectins (MBLs). These bind to certain arrangements of multiple
mannose molecules that characterize microbial cells. Once an
MBL attaches to a surface, it can interact with other complement
system components to form a C3 convertase. mannose, p. 31

Classical Pathway
Complement system activation by the classical pathway requires
antibodies, a component of adaptive immunity. When antibodies
bind to an antigen (forming an antigen-antibody complex, also
called an immune complex), they interact with the same comple-
ment system component involved with the lectin pathway to form
a C3 convertase.

Effector Functions
of the Complement System
Activation of the complement system eventually leads to three
major protective outcomes: opsonization, an inflammatory
response, and lysis of foreign cells (figure 14.10).

Opsonization FIGURE 14.11 Membrane Attack Complexes (MACs) in a Cell

The C3b concentration increases substantially when the comple- Membrane Each dark dot in this electron micrograph is a MAC.
ment system is activated, and these molecules bind to bacterial ? How do MACs cause cells to lyse?
346 Chapter 14 The Innate Immune Response

a Host cell surface

C3b is quickly inactivated when it attaches to the surface.

Complement Other C3b attaches to host cell

regulatory complement surface; complement regulatory
protein proteins proteins inactivate it.
C3b
C3 convertase not formed;
complement system not
activated.
Host cell surface

b Microbial surface
C3b remains active when it attaches to the surface.

Complement Other C3b attaches to bacterial surface. Other complement proteins

regulatory complement attach to C3b bound to the
protein proteins surface, forming a C3
convertase.
Complement
system
C3b activated.

Microbial cell surface

FIGURE 14.12 Complement Regulatory Proteins Inactivate C3b (a) Molecules in host cell membranes bind regulatory proteins that
quickly inactivate C3b. (b) Most microbial cell surfaces do not bind the regulatory proteins, so they trigger the alternative pathway of
complement activation.
? Some pathogens attract complement regulatory proteins to their surfaces. How would this help the pathogens avoid destruction?

Regulation of the Complement System 14.7 ■ Phagocytosis

A number of different control mechanisms prevent host cells from
activating the complement system and also protect them from Learning Outcomes
the effector functions of the complement proteins. For example, 7. Outline the steps of phagocytosis.
molecules in host cell membranes bind regulatory proteins that 8. Compare and contrast the roles of macrophages and neutrophils.
quickly inactivate C3b. This prevents host cell surfaces from
triggering the alternative pathway of complement regulation Phagocytes routinely engulf and digest material, including invad-
(figure 14.12). It also prevents host cells from being opsonized. ing organisms. In routine situations, such as when microbes enter
Most microbial cell surfaces do not bind the regulatory proteins, through a minor skin wound, resident macrophages in the tissues
but as we will discuss in chapter 16, some pathogens have mecha- destroy the relatively few invaders that enter. If the microbes are
nisms to hijack the host’s protective mechanisms. not rapidly cleared, macrophages produce cytokines to recruit
additional phagocytes—particularly neutrophils—for extra help.
MicroAssessment 14.6
The complement system can be activated by three different pathways The Process of Phagocytosis
that each lead to opsonization, an inflammatory response, and lysis of
Phagocytosis involves a series of steps (figure 14.13). These are
foreign cells.
particularly important medically, because most pathogens have
16. What role does C3b play in opsonization and complement system evolved the ability to evade one or more of them, a topic explored
activation?
in chapter 16.
17. The body’s own cells do not trigger the alternative pathway of
complement pathway activation. How come?
Chemotaxis 1
18. From a pathogen’s standpoint, why would it be beneficial to bind
regulatory proteins that inactivate C3b? + Phagocytic cells are recruited to the site of infection or tissue
damage by chemicals that act as chemoattractants. These include
 Part III Microorganisms and Humans 347

1 Chemotaxis C5a

Microbes

C3b
Phagocyte
Lysosomes
2 Recognition
and attachment
6 Exocytosis
Pseudopod Phagosome
C3b
Phagolysosome
C3b receptors
on phagocyte

Digestive
enzymes

5 Destruction
4 Phagosome maturation and and digestion
3 Engulfment phagolysosome formation

FIGURE 14.13 Phagocytosis This diagram shows a microbe that has been opsonized by the complement protein C3b; certain classes of
antibodies can also function as opsonins.
? What would happen if a bacterium could prevent the phagosome from fusing with lysosomes?

products of microorganisms, phospholipids released by injured host various endosomes, allowing it to gain properties that character-
cells, chemokines, and the complement system component C5a. ize those endosomes. For example, the pH becomes progressively
more acidic. Finally, the phagosome fuses with enzyme-filled
Recognition and Attachment 2 lysosomes, forming a phagolysosome. lysosome, p. 77

Phagocytic cells use various receptors to bind invading microbes The phagosome maturation stages are highly regulated, and
either directly or indirectly. For example, direct binding occurs appear to depend on the type of material ingested. For instance, a
when a phagocyte’s receptors bind mannose, a sugar found on phagosome that contains host cell material has a different fate than
some bacteria and yeasts. Indirect binding happens when a particle one that carries microbial components.
has first been opsonized. Opsonins are secreted proteins that tag
particles for phagocytosis and include the complement component Destruction and Digestion 5
C3b and certain classes of antibody molecules. Phagocytes have A number of factors within the phagolysosome work together
specific receptors for opsonins, making it easier for the cells to to destroy an engulfed invader. O2 consumption increases
attach to and subsequently engulf the material. dramatically—a phenomenon called respiratory burst—allowing
an enzyme to produce reactive oxygen species (ROS), which
Engulfment 3 are toxic. Another enzyme makes nitric oxide, which reacts with
Once the phagocyte has attached to a particle, it sends out pseu- ROS to produce additional toxic compounds. Special pumps move
dopods that surround and engulf the material. This action brings protons into the phagolysosome, lowering the pH. The various
the material into the cell, enclosed in a phagosome. If a phagocyte enzymes contributed by the lysosomes degrade peptidoglycan and
encounters something too large to engulf, it releases its toxic con- other components. Defensins damage membranes of the invader,
tents as a means of destroying it. pseudopod, p. 72 phagosome, p. 72 and lactoferrin ties up iron. reactive oxygen species, p. 90

Phagosome Maturation Exocytosis 6

and Phagolysosome Formation 4 Following digestion, the vesicle fuses with the plasma membrane,
Initially, a phagosome has no antimicrobial capabilities, but it expelling the remains. In the case of macrophages, some of the
matures to develop these. As part of this process, it fuses with ingested material is also put on the cell’s surface as a way of
348 Chapter 14 The Innate Immune Response

displaying bits of invaders to certain cells of the adaptive immune NET ensnare microbes, allowing the granule contents (enzymes
system. Details of this process will be discussed in chapter 15. and peptides) that accumulate within the NET to destroy them.
exocytosis, p. 72
MicroByte
For every neutrophil in the circulatory system, about 100 more are
waiting in the bone marrow, ready to be mobilized when needed.
Specialized Attributes of Macrophages
Macrophages can be viewed as the scavengers and sentries—
routinely phagocytizing dead cells and debris, but ready to destroy MicroAssessment 14.7
invaders and call in reinforcements when needed. They are always The process of phagocytosis includes chemotaxis, recognition and
present in tissues, where they either slowly wander or remain attachment, engulfment, phagosome maturation and phagolysosome
stationary. These phagocytic cells play an essential role in every formation, destruction and digestion, and exocytosis. Macrophages
major tissue in the body. are long-lived and always present in tissues; they can be activated to
Macrophages live for weeks to months, and maintain their enhance their killing power. Neutrophils are highly active, short-lived
phagocytic cells that must be recruited to the site of damage.
killing power by continually regenerating their lysosomes. As
macrophages die, circulating monocytes—which can differentiate 19. How does a phagolysosome differ from a phagosome?
into macrophages—leave the blood and migrate to the tissues to 20. Tuberculosis is characterized by granulomas called tubercles.
replace them. Monocyte migration increases in response to inva- What is a granuloma?
sion and tissue damage. 21. What could a microorganism do to avoid engulfment? +
Macrophages have several important characteristics that
allow them to carry out their diverse tasks. Various TLRs on their
surfaces and in phagosomes help them sense microbial invaders. 14.8 ■ The Inflammatory
When these receptors are triggered, the macrophage responds by
producing cytokines that alert and stimulate various other cells Response
of the immune system. Macrophages can increase their otherwise
Learning Outcomes
limited killing power to become activated macrophages. One
mechanism for this requires the assistance of certain T cells, 9. Describe the inflammatory process, focusing on the factors that
initiate the response and the outcomes of inflammation.
an example of the cooperation between the innate and adaptive
defenses. The compounds produced by activated macrophages 10. Compare and contrast apoptosis and pyroptosis.
also damage tissues when released, so it would be potentially
harmful for macrophages to routinely be in an activated state. When tissues have been damaged, such as when an object pen-
Details of the activation process, including the roles of T cells, etrates the skin or microbes are introduced, inflammation occurs.
will be discussed in chapter 15.  macrophage activation, p. 373 The purpose of this is to contain a site of damage, localize the
If activated macrophages fail to destroy microbes, the phago- response, eliminate the invader, and restore tissue function.
cytes can fuse together to form giant cells. Macrophages, giant Everyone has experienced the signs of inflammation; in fact, the
cells, and T cells form concentrated groups called granulomas Roman physician Celsus described the four cardinal signs in the
that wall off and retain organisms or other material that cannot be first century a.d.—swelling, redness, heat, and pain. A fifth sign,
destroyed; again, this is an example of the cooperation between loss of function, is sometimes present.
defense systems. Granulomas, which are part of the disease
process in tuberculosis and several other illnesses, prevent the Factors That Trigger
microbes from escaping to infect other cells (see figure  21.20).
Unfortunately, they also harm the host because they interfere with
an Inflammatory Response
normal tissue function.  tuberculosis, p. 502 A number of factors trigger an inflammatory response, but they
often involve the pattern recognition receptors described in sec-
tion 14.5—particularly TLRs and NLRs. Recall that these recep-
Specialized Attributes of Neutrophils tors detect pathogen-associated molecular patterns (PAMPs)
and damage-associated molecular patterns (DAMPs). The trig-
Neutrophils can be viewed as the rapid response team—quick to
gers of inflammation cause host cells to release inflammatory
move into an area of trouble and ready to eliminate the invaders.
mediators, a collective term for various pro-inflammatory
They play a critical role during the early stages of inflammation,
cytokines and chemicals such as histamine and bradykinin.
being the first cell type recruited to the site of damage from the
Inducers of the inflammatory process include:
bloodstream. They have more killing power than macrophages.
The cost for their effectiveness, however, is a relatively short ■ Microbes. When TLRs on sentinel cells such as macrophages
life span of only 1 to 2 days in the tissues; once they have used detect PAMPs, the cells produce inflammatory mediators.
their granules, they die. Fortunately, many more neutrophils are One of these, tumor necrosis factor (TNF), induces the liver
in reserve. to synthesize acute-phase proteins, a group of proteins that
Neutrophils not only kill microbes through phagocytosis, they facilitate phagocytosis and complement activation. When
also release the contents of their granules along with DNA to form NLRs within cells detect PAMPs, additional inflamma-
neutrophil extracellular traps (NETs). The DNA strands in the tory mediators are released. Meanwhile, microbial surfaces
 Part III Microorganisms and Humans 349

trigger complement activation, also leading to an inflamma- vessels, and the migration of leukocytes out of the bloodstream
tory response. and into the tissues (figure 14.14).
■ Tissue damage. The sensors of DAMPs are not well under- The diameter of local blood vessels increases during inflam-
stood, but seem to involve NLRs. As with detection of mation due to the action of inflammatory mediators. This results
PAMPs, these cause cells to release inflammatory mediators. in greater blood flow to the area, causing the heat and redness
If blood vessels are injured, two enzymatic cascades are acti- associated with inflammation. It also slows the blood flow in
vated. One is the coagulation cascade, which results in blood the capillaries. Because of the dilation, normally tight junctions
clotting, and the other produces several molecules such as between endothelial cells are disrupted, allowing more fluid
bradykinin, which increases blood vessel permeability. to leak from the vessels and into the tissue. This fluid contains
various substances such as transferrin, complement system pro-
teins, and antibodies that help counteract invading microbes.
The Inflammatory Process The increase of fluids in the tissues causes the swelling and pain
The inflammatory process involves a cascade of events that result associated with inflammation. The direct effects of chemicals on
in dilation of small blood vessels, leakage of fluids from those sensory nerve endings also cause pain (see Perspective 14.1).

Pro-
inflammatory
chemicals
Bacteria

Inflammatory Phagocytic
mediators are cells destroy Neutrophil
Resident released in and remove
macrophage response to invaders.
Blood vessel microbial Neutrophil Recruited
components macrophage
and tissue
damage.

Monocyte

Normal blood flow in the tissues Neutrophils are the first phagocytes As the infection is brought under
as the injury occurs. recruited to the site. control, macrophages ingest dead
cells and debris.
(a)

Tighter Diapedesis
Loose adherence; adherence
cells tumble to a halt.

Inflammatory mediators cause small blood vessels to dilate. The phagocytic cells tumble to
a halt and then squeeze between the endothelial cells and enter the tissues.
(b)

FIGURE 14.14 The Inflammatory Response (a) The process of inflammation. (b) Phagocytes leave the blood vessels and move to the
site of infection.
? Which type of phagocyte is the first to be recruited to a site of inflammation?
350 Chapter 14 The Innate Immune Response

PERSPECTIVE 14.1
For Schistosoma, the Inflammatory Response Delivers
The parasitic flatworms that cause schisto- with the aid of digestive enzymes; schisto- a manner similar to what is experienced as a
somiasis do not shy from the immune response somes are rare among pathogens because they sliver in the skin works its way to the surface.
when it comes to reproducing; instead they can penetrate intact skin. The larvae then enter Only half of the ova are expelled, however,
use it to move their ova to an environment the bloodstream where the parasite can live for and the remaining ones are often swept by the
where they might hatch. Adult females of over a quarter of a century. bloodstream to the liver. The inflammatory pro-
Schistosoma species, which live in the blood- Schistosoma species have separate sexes cess and granuloma formation there gradually
stream of infected hosts, lay their ova in veins and, remarkably, the male and female worms destroy liver cells, replacing the cells with scar
near the intestine or bladder. They then rely locate one another in the bloodstream. The tissue. Malfunction of the liver results in mal-
on the inflammatory response there to expel male’s body has a deep longitudinal groove nutrition and a buildup of pressure in intestinal
the ova, completing one portion of a complex in which he clasps his female partner to live and esophageal veins. Fluid accumulates in the
life cycle. The ova released in feces or urine in copulatory embrace (schistosoma means abdominal cavity and hemorrhage occurs if the
can hatch—liberating a larval stage called “split-body,” referring to the long slit). The esophageal veins rupture.
a miracidium—if untreated sewage reaches adult worms effectively mask themselves from Despite their complex life cycle, Schistosoma
water. The miracidium then infects a specific the immune system by adsorbing various species are highly successful parasites. Not
freshwater snail host, where it multiplies asex- blood proteins; this provides them with a only are they adept at avoiding certain immune
ually. The infected snail then releases large primitive stealth “cloaking device.” responses that would otherwise lead to their
numbers of another larval form, cercariae, The worms migrate to the veins of either destruction, they have learned to exploit the
which swim about in search of a human host. the intestine or bladder (depending on the schis- inflammatory response for their own spread.
The parasite is acquired when a person tosome species) to lay hundreds of ova per day. Over 200 million people worldwide are
wades or swims in contaminated water. The cer- The innate immune system responds vigorously infected with these parasites, resulting in over
cariae penetrate the skin by burrowing through to the highly antigenic eggs, ejecting them in 500,000 deaths each year.

Some of the pro-inflammatory cytokines cause endothelial giant cells accumulate, and granulomas form.   giant cells, p.  373
cells in the local area to produce adhesion molecules that loosely  granulomas, p. 373
“grab” phagocytes. The phagocytes normally flow rapidly through
the vessels but slowly tumble to a halt as the adhesion molecules
attach. The phagocytic cells themselves begin producing a dif- Damaging Effects of Inflammation
ferent type of adhesion molecule that strengthens the attachment. The inflammatory process can be compared to a sprinkler system
Then, in response to various chemoattractants, the phagocytes that prevents fire from spreading in a building. Although the
leave the blood vessels and move into the area. They do this by process usually limits damage and restores function, the response
squeezing between the cells of the dilated vessel, a process called itself can cause significant harm. One undesirable consequence is
diapedesis. Neutrophils are the first to arrive at the site of infec- that some enzymes and toxic products contained within phago-
tion, and they actively phagocytize foreign material. Monocytes cytic cells are inevitably released, damaging tissues.
(which mature into macrophages at the site of infection) and If inflammation is limited, such as in a response to a cut
lymphocytes arrive later. Clotting factors in the fluid that leaks finger, the damage caused by the process is normally minimal.
into the tissues initiate clotting reactions in the surrounding area, If the process occurs in a delicate system, however, such as the
walling off the site of infection. This helps prevent bleeding and membranes that surround the brain and spinal cord, the conse-
stops spread of invading microbes. As the inflammatory process quences can be much more severe, even life-threatening. As you
continues, large quantities of dead neutrophils accumulate. Those learn more about infectious diseases, you will notice that many of
dead cells, along with tissue debris, make up pus. A large amount the most severe effects of infection result from the inflammatory
of pus constitutes an abscess (see figure 22.2). abscess, p. 550 response.
The extent of inflammation varies, depending on the nature of
the injury, but the response is localized, begins immediately upon
injury, and increases rapidly. A short-term inflammatory response
Cell Death and the
is called acute inflammation and is marked by a prevalence of Inflammatory Process
neutrophils. As the infection is brought under control, resolution In addition to traumatic cell death that results from tissue dam-
of inflammation begins. Neutrophils stop entering the area and age, host cells can self-destruct. This capability allows the host to
macrophages clean up the damage by ingesting dead cells and eliminate any cells no longer needed, and it serves as a mechanism
debris. As the area heals, new capillaries grow, destroyed tissues for sacrificing “self” cells that might otherwise spread an infec-
are replaced, and scar tissue forms. tion. One type of programmed cell death avoids an inflammatory
If acute inflammation cannot limit the infection, chronic response, whereas another type promotes one.
inflammation occurs. This is a long-term inflammatory process Apoptosis (apo means “off”; ptosis means “falling”) is a
that can last for years. In chronic inflammation, macrophages and programmed cell death that does not trigger an inflammatory
 Part III Microorganisms and Humans 351

response. During apoptosis, the dying cells undergo certain regulating center “sets” the body’s thermostat at a higher level. An
changes. For example, the shape of the cell changes, enzymes cut oral temperature above 37.8°C is regarded as fever.
the DNA, and portions of the cell bud off, effectively shrinking the A higher temperature setting occurs as a result of certain
cell. Some changes appear to signal macrophages that the remains pro-inflammatory cytokines released by macrophages when they
of the cell are to be engulfed without the events associated with detect microbial products. The cytokines are carried in the blood-
inflammation. stream to the brain, where they act as messages that microbes
If the pattern recognition receptors in a macrophage’s cyto- have invaded the body. These cytokines and other fever-inducing
plasm are triggered, that cell might initiate pyroptosis ( pyro substances are pyrogens. Fever-inducing cytokines are called
means “fire”). This programmed self-destruction triggers an endogenous pyrogens, indicating the body makes them, whereas
inflammatory response, recruiting various components of the microbial products are called exogenous pyrogens, indicating they
immune system to the region. are introduced from external sources. The temperature-regulating
center responds to pyrogens by raising body temperature.
MicroAssessment 14.8 The adverse effect of fever on pathogens partly involves ideal
The inflammatory response is initiated when microbes invade or
growth temperature. Bacteria that grow best at 37°C are less likely
tissues are damaged. The outcome is dilation of small blood vessels, to cause disease in people with fever because bacterial growth
leakage of fluids from those vessels, and migration of leukocytes out rates often decline sharply above their optimum growth tempera-
of the bloodstream and into the tissue. Inflammation helps contain an ture. A slower growth rate allows more time for other defenses to
infection, but the response itself can be damaging. Apoptosis destroys destroy invaders. temperature requirements, p. 89
“self” cells without initiating inflammation; pyroptosis triggers an A moderate rise in temperature increases the rate of enzy-
inflammatory response. matic reactions. Fever has been shown to enhance the inflamma-
22. Describe two general events that can initiate inflammation. tory response, phagocytic killing by leukocytes, multiplication of
23. Describe two changes in cells undergoing apoptosis. lymphocytes, release of substances that attract neutrophils, and
24. Infection of the fallopian tubes can lead to infertility. Why would production of interferons and antibodies. Release of leukocytes
this be so? + into the blood from the bone marrow also increases.

MicroAssessment 14.9
Fever results when macrophages release pro-inflammatory cytokines;
14.9 ■ Fever this occurs when the TLRs on the macrophages are engaged by
microbial products.
Learning Outcome
25. What are endogenous and exogenous pyrogens?
11. Describe the induction and outcomes of fever.
26. How does fever inhibit the growth of pathogens?
Fever is an important host defense mechanism, and a strong indica- 27. Syphilis was once treated by infecting the patient with the
parasite that causes malaria, a disease characterized by
tion of infectious disease, especially of bacterial origin. The tem-
repeated bouts of fever, shaking, and chills. Why would this
perature within the human body is normally kept around 37°C by a treatment cure syphilis? +
temperature-regulation center in the brain. During an infection, the

Summary
14.1 ■ Overview of the Innate Defenses Normal Microbiota (Flora)
First-line defenses prevent entry, sensor systems detect invasion, and Members of the normal microbiota competitively exclude pathogens
effector mechanisms destroy and remove the invader (figures 14.1). and stimulate the host defenses.
Cells have pattern recognition receptors (PRRs) that recognize
invaders. The complement system is activated in response to certain
14.3 ■ The Cells of the Immune System (figure 14.5, table 14.1)
invaders. The inflammatory response involves many components of
innate immunity, including phagocytes. Granulocytes
Granulocytes include neutrophils, basophils, and eosinophils.
14.2 ■ First-Line Defenses (figure 14.2)
Mononuclear Phagocytes
Physical Barriers
Monocytes circulate in blood; macrophages are in tissues (figure 14.6).
The skin is composed of two main layers—the dermis and the epider-
mis. Mucous membranes are constantly bathed with mucus and other Dendritic Cells
secretions that help wash microbes from the surfaces (figure 14.3). Dendritic cells develop from monocytes; some have other origins.
Antimicrobial Substances Lymphocytes
Lysozyme, peroxidase enzymes, lactoferrin, and defensins inhibit or Lymphocytes, which include B cells, T cells, and natural killer (NK)
kill microorganisms (figure 14.4). cells, are involved in adaptive immunity.
352 Chapter 14 The Innate Immune Response

14.4 ■ Cell Communication Specialized Attributes of Macrophages

Surface Receptors
Macrophages are always present in tissues to some extent but can
call in reinforcements when needed. A macrophage can become
Surface receptors bind ligands, allowing the cell to detect substances.
an activated macrophage. Macrophages, giant cells, and T cells
Cytokines (table 14.2) form granulomas that wall off and retain material that cannot be
Cytokines include chemokines, colony-stimulating factors (CSFs), destroyed.
interferons (IFNs), interleukins (ILs), and tumor necrosis factor (TNF). Specialized Attributes of Neutrophils
Adhesion Molecules Neutrophils are the first cell type recruited from the bloodstream to the
Adhesion molecules allow cells to adhere to other cells. site of damage.

14.5 ■ Pattern Recognition Receptors (PRRs) 14.8 ■ The Inflammatory Response

Swelling, redness, heat, and pain are the signs of inflammation, the
Toll-Like Receptors (TLRs) body’s attempt to contain a site of damage, localize the response, elimi-
TLRs allow sentinel cells to detect extracellular molecules that signify nate the invader, and restore tissue function.
the presence of microbes (figure 14.7).
Factors That Trigger an Inflammatory Response
NOD-Like Receptors (NLRs) Inflammation is initiated when microbes are detected by TLRs, NLRs,
NLRs allow cells to detect internal invaders or cell damage (figure 14.8). or the complement system, or when tissue damage occurs.
RIG-Like Receptors (RLRs) The Inflammatory Process
RLRs allow cells to detect viral nucleic acid. Virally infected cells The inflammatory process results in dilation of small blood vessels,
respond by making interferons, causing nearby cells to prepare to leakage of fluids from those vessels, and movement of leukocytes from
undergo apoptosis if they become infected with a virus (figure 14.9). the bloodstream into the tissues (figure  14.14). Acute inflammation is
marked by the prevalence of neutrophils; chronic inflammation is
14.6 ■ The Complement System characterized by macrophage and giant cell accumulation, and granu-
loma formation.
Complement System Activation
The complement system detects microbial cells and antibodies bound to Damaging Effects of Inflammation
antigens, and is activated in response (figure 14.10). The inflammatory process can be damaging to the host, and in some
cases this is life-threatening.
Effector Functions of the Complement System
The major protective outcomes of complement system activation Cell Death and the Inflammatory Process
include opsonization, an inflammatory response, and lysis of foreign Apoptosis is a mechanism of eliminating “self” cells without trigger-
cells (figure 14.11). ing an inflammatory response; pyroptosis triggers an inflammatory
response.
Regulation of the Complement System
Complement regulatory proteins prevent host cell surfaces from activat- 14.9 ■ Fever
ing the complement system via the alternative pathway (figure 14.12). Fever results when macrophages release certain pro-inflammatory cyto-
kines. It inhibits the growth of many pathogens and increases the rate
14.7 ■ Phagocytosis
of various body defenses.
The Process of Phagocytosis (figure 14.13)
The steps of phagocytosis include chemotaxis, recognition and attach-
ment, engulfment, phagosome maturation and phagolysosome forma-
tion, destruction and digestion, and exocytosis.

Review Questions
Short Answer Multiple Choice
1. Describe how the skin protects against infection. 1. Lysozyme does which of the following?
2. What factors in saliva aid in protection against microbes? a) Disrupts cell membranes
3. Why is iron metabolism important in body defenses? b) Hydrolyzes peptidoglycan
c) Waterproofs skin
4. Name two categories of cytokines and give their effects.
d) Propels gastrointestinal contents
5. What is the function of a TLR?
e) Propels the cilia of the respiratory tract
6. Contrast the pathways of complement activation. 2. The hematopoietic stem cells in the bone marrow can become
7. How do complement proteins cause foreign cell lysis? which of the following cell types?
8. How do phagocytes enter tissues during an inflammatory 1. Red blood cell 2. T cell 3. B cell
response? 4. Monocyte 5. Macrophage
9. How is acute inflammation different from chronic inflammation? a) 2, 3 b) 2, 4 c) 2, 3, 4, 5
10. Describe the function of apoptosis. d) 1, 4, 5 e) 1, 2, 3, 4, 5
 Part III Microorganisms and Humans 353

3. All of the following refer to the same type of cell except 10. Which of the following statements about inflammation is false?
a) macrophage. b) neutrophil. a) Vasodilation results in leakage of blood components.
c) poly. d) PMN. b) The process can damage host tissue.
4. TLRs are triggered by all of the following compounds except c) Neutrophils are the first to migrate to a site of inflammation.
a) peptidoglycan. b) glycolysis enzymes. d) Apoptosis induces inflammation.
c) lipopolysaccharide. d) flagellin. e) The signs of inflammation are redness, swelling, heat, and pain.
e) certain nucleotide sequences.
Applications
5. The direct/immediate action of interferon on a cell is to
1. Paraplegic patients often have recurrent urinary tract infections.
a) interfere with the replication of the virus.
Why would the condition keep coming back in spite of repeated
b) prevent the virus from entering the cell. treatment?
c) stimulate synthesis of inactive “suicide enzymes.”
2. A cattle farmer sees a sore on the leg of one of his cows. The
d) stimulate the immune response. farmer feels the sore and notices that the area just around the
e) stop the cell from dividing. sore is warm to the touch. A veterinarian examines the wound
6. A pathogen that can avoid the complement component C3b would and explains that the warmth may be due to inflammation. The
directly protect itself from farmer wants an explanation of the difference between the local-
a) opsonization. b) triggering inflammation. ized warmth and fever. What would be the vet’s explanation to the
c) lysis. d) inducing interferon. e) antibodies. farmer?
7. Which of the following statements about phagocytosis is false?
Critical Thinking +
a) Phagocytes move toward an area of infection by chemotaxis.
1. A student argues that phagocytosis is a wasteful process because
b) Digestion of invaders occurs within a phagolysosome.
after engulfed organisms are digested and destroyed, the remain-
c) Phagocytes have receptors that recognize C3b bound to
ing material is excreted from the cell (see figure 14.13). A more
bacteria.
efficient process would be to release the digested material inside
d) Phagocytes have receptors that recognize antibodies bound to the cell. This way, the material and enzymes could be reused by the
bacteria. cell. Does the student have a valid argument? Why or why not?
e) Macrophages die after phagocytizing bacteria, but neutrophils
2. According to figure 14.9, any cell infected by viruses may die due
regenerate their lysosomes and survive.
to the action of interferons. This strategy, however, seems counter-
8. All of the following cell types are found in a granuloma except productive. The same result would occur without interferon—any
a) neutrophils. b) macrophages. cell infected by a virus might die directly from the virus. Is there
c) giant cells. d) T cells. any apparent benefit from the interferon action?
9. All of the following trigger an inflammatory response except
a) engagement of TLRs.
b) complement system activation.
c) interferon induction of antiviral protein synthesis.
d) tissue damage.
15 The Adaptive Immune Response
KEY TERMS
Adaptive Immunity Protection
provided by immune responses that
mature throughout life; involves
B cells and T cells.
macrophages, and assist other parts
of the adaptive immune response.
Humoral Immunity Immunity
involving B cells and an antibody
Antibody Y-shaped protein that response.
binds antigen. Lymphocytes A group of white
Antigen Molecule that reacts blood cells (leukocytes) that have
specifically with either an antibody antigen-specific receptors on their
or an antigen receptor on a surface; includes B cells and T cells.
lymphocyte. Major Histocompatibility
Antigen-Presenting Cells (APCs) Complex (MHC) Molecules Cell
Cells such as B cells, macrophages, surface molecules that present
and dendritic cells that can present antigen to T cells; includes MHC
exogenous antigens to naive or class I molecules and MHC class II
memory T cells, activating them. molecules.
B Cell Type of lymphocyte Memory Lymphocytes Long-
programmed to make antibodies. lived descendants of activated
lymphocytes that can quickly
Cell-Mediated Immunity (CMI)
respond when a specific antigen is
Immunity involving a T-cell
encountered again.
response.
Plasma Cell Effector form of a
Clonal Selection Process in which
B cell; it functions as an antibody-
a lymphocyte’s antigen receptor
Blood clot, with erythrocytes, fibrin filaments, and a lymphocyte (color- secreting factory.
enhanced scanning electron micrograph).
binds to an antigen, allowing the
lymphocyte to proliferate. TC Cell Effector form of a cytotoxic
T cell; it induces apoptosis in
Cytotoxic T Cell Type of
infected or cancerous “self” cells.
lymphocyte programmed to destroy
A Glimpse of History infected or cancerous “self” cells. TH Cell Effector form of a helper
T cell; it activates B cells and
Dendritic Cell Cell type
Near the end of the nineteenth century, diphtheria was a terrifying macrophages, and releases cytokines
responsible for activating naive
disease that killed many infants and small children. The first symp- T cells.
that stimulate other parts of the
tom was a sore throat, often followed by the development of a gray immune system.
Helper T Cell Type of lymphocyte
membrane that could come loose and block the airway. Frederick
programmed to activate B cells and
Loeffler, working in Robert Koch’s laboratory in Berlin, found club-
shaped bacteria growing in the throats of people with the disease but
not elsewhere in their bodies. He hypothesized that the organisms
were making a poison that spread through the bloodstream. In Paris,
at the Pasteur Institute, Emile Roux and Alexandre Yersin followed
up by growing the bacteria and extracting the poison, or toxin, from disease. The substances with antitoxin properties were then given the
culture fluids. When the toxin was injected into guinea pigs, it gener- name antibodies, and materials that induced antibody production were
ally killed them. Alexandre Yersin, p. 670 called antigens.
Back in Berlin, Emil von Behring injected diphtheria toxin into Emil von Behring received the first Nobel Prize in Medicine in 1901
guinea pigs that had recovered from lab-induced diphtheria. These for his work on antibody therapy. It took many more decades of inves-
animals did not become ill, suggesting that something in their blood tigation to reveal the biochemical nature of antibodies. In 1972, Rodney
protected them; von Behring called it antitoxin. To test this idea, he Porter and Gerald Edelman were awarded a Nobel Prize for their part in
mixed toxin with serum (the liquid portion of blood) from a guinea pig determining the structure of antibodies.
that previously had diphtheria, and injected it into one that had not had
the disease. The animal remained well. In further experiments, he cured

I
animals with diphtheria by giving them antitoxin. n contrast to the innate immune response, which is always
The effectiveness of antitoxin was put to the test in late 1891, when ready to respond to patterns that signify invasion or dam-
a diphtheria epidemic broke out in Berlin. On Christmas night, antitoxin age, the adaptive immune response matures throughout life,
was first given to an infected child, who then recovered from the dreaded developing from the immune system arsenal the most effective

354
 Part III Microorganisms and Humans 355

means to eliminate a specific invader when it is encountered. 15.1 ■ Strategy of the Adaptive
The protection provided by the response is called adaptive
immunity. On first exposure to a given microbe, adaptive Immune Response
immunity takes a week or more to build. During this delay, the
host depends on innate immunity for protection, which may not Learning Outcome
be sufficient to prevent disease. In some cases, a person may 1. Compare and contrast the general aspects of humoral immunity
not survive long enough for the adaptive immune response to and cell-mediated immunity.
reach an effective level.
An important characteristic of adaptive immunity is This first response to a particular antigen is called the primary
memory, a stronger response to re-exposure. Individuals who response. As a result of this initial encounter, the adaptive
survive diseases such as measles, mumps, or diphtheria gener- immune system “remembers” the mechanism that proved effec-
ally never develop the same disease again. Vaccination now tive against that specific antigen. As a result, when the same anti-
prevents these diseases by exposing a person’s immune system gen is encountered later in life, there is a stronger antigen-specific
to harmless forms of the pathogen or its products. It is true immune response called the secondary response. antigen, p. 335
that diseases such as influenza can be contracted repeatedly, The adaptive immune response uses two basic strategies for
but that is generally due to the pathogen’s ability to continu- countering foreign material. One response, humoral immunity,
ally evade the host defenses, a topic discussed in chapter 16. works to eliminate extracellular antigens—for example, bacteria,
vaccination, p. 421
toxins, or viruses in the bloodstream or in tissue fluids. The other,
The adaptive immune response has molecular specificity. A called cell-mediated immunity (CMI), or cellular immunity,
response that protects an individual from developing measles does deals with antigens residing within a host cell, such as a virus
not prevent a different disease—for example, chickenpox. infecting a cell. Humoral and cell-mediated immunity are both
Another important aspect of adaptive immunity is tolerance, powerful and, if misdirected, can damage the body’s own tis-
the ability to ignore any given molecule. Most significantly, the sues. Because of this, the adaptive immune response is tightly
immune system can distinguish normal host cells from invading regulated; before a lymphocyte can unleash its power, it generally
microbes, an ability referred to as self versus non-self recognition. requires confirmation from a different cell type that the antigen
A more accurate description might be “healthy self” versus “dan- does indeed represent a threat.
gerous,” the latter including invading microbes as well as cancer-
ous or other “corrupt” cells. Regardless, the ability to develop Overview of Humoral Immunity
self-tolerance is critical because without it the immune system B lymphocytes, or B cells, are responsible for humoral
would routinely turn against the body’s own cells, attacking them immunity (figure 15.1). Their name reflects the fact that they
just as it does invading microbes. Self-tolerance is not a fail-safe develop in an organ in birds called the bursa. In humans, how-
system, however, which is why autoimmune diseases can occur. ever, B cells develop in the bone marrow.
 autoimmune disease, p. 412 In response to extracellular antigens, B cells may be triggered
The adaptive immune system is extraordinarily complex, to proliferate and then differentiate into plasma cells, which func-
involving an intricate network of cells, cytokines, and other tion as factories that produce Y-shaped proteins called antibodies.
compounds. The need for such complexity is logical given These molecules bind to antigens, providing protection by mech-
the task of the system, but it makes the system difficult to anisms described shortly. A high degree of specificity is involved
explain and comprehend! Although it might seem that any in the binding, so many different antibody molecules are needed
description should start at the beginning and then continue in to bind to the wide array of antigens encountered throughout life.
a linear progression, that approach can be bewildering. As an Some of the B cells form memory B cells, long-lived cells that
analogy, imagine explaining how to build a car to someone respond more quickly if the antigen is encountered again.
who has never even seen an automobile, much less driven Antibody molecules have two functional regions—the two
one; the person would have no idea why certain parts were identical arms and the stem of the molecule. The arms bind spe-
being put together. With that in mind, we have chosen to start cific antigen molecules; the amino acid sequence of the end of the
the chapter with a general overview of the adaptive immune arms varies among antibodies, providing the basis for their speci-
response, focusing on the essential elements of the process ficity. The stem functions as a “red flag,” tagging antigen bound
and how the response eliminates invading microbes. Armed by antibodies and enlisting other immune system components to
with this foundation, it should then be easier for you to fol- eliminate the molecule.
low the more in-depth descriptions of the various parts and Antibodies protect the host by both direct and indirect mech-
how they fit together. At the end of the chapter, we will focus anisms. The direct effect is due to their ability to bind antigens.
on the development of the system, concentrating on how the Simply by binding, they can coat an antigen, thereby preventing it
lymphocytes—the primary participants in the adaptive immune from attaching to a host cell. For example, an antibody-coated viral
responses—gain the specificity required to respond to an particle cannot attach to its receptor, and therefore cannot enter the
incredibly diverse and ever-changing assortment of microbes. cell. The indirect protective effect is due to the “red flag” region
lymphocytes, p. 340 tagging the antigen for elimination. attachment of viruses, p. 317
 Innate immunity
Dendritic cell

Activates T cells that

bind antigens
Activation representing “danger”

Naive B cell Naive helper T cell Naive cytotoxic T cell

Proliferation
and differentiation
Plasma cells
TH cells TC cells

Produce Deliver Deliver “death

antibodies cytokines packages”

Effector action
and consequence

Macrophage that Macrophage with Infected Infected “self”

Antibodies Antibodies bind has engulfed increased killing power “self” cell cell undergoes
antigen invaders apoptosis
Adaptive immunity Adaptive immunity
(humoral) (cell-mediated)

FIGURE 15.1 Overview of Humoral and Cell-mediated Immunity Humoral immunity protects against antigens in blood and tissue fluid
(extracellular antigens); cell-mediated immunity protects against antigens within host cells (intracellular antigens). In this diagram, solid arrows represent
the path of a cell or molecule; dashed arrows represent a cell’s interactions and effector functions; antigen receptors and memory cells are not shown.

? How do antibodies protect against infection?

How does a naive B cell (one that has never “seen” antigen T cells and helper T cells. Both of these have multiple copies of a
before) know when to respond? The B-cell surface has numer- surface molecule called a T-cell receptor (TCR), which is func-
ous copies of a B-cell receptor (BCR), a membrane-bound tionally analogous to a BCR; it allows the cell to bind a specific
version of the specific antibody the B cell is programmed to antigen (figure 15.2). Unlike a BCR, however, a TCR does not
make (figure 15.2). If a naive B cell encounters an antigen recognize free antigen. Instead, the antigen must be presented by
that its BCR binds, the cell is triggered to multiply. Some one of the body’s own cells.
of the resulting clones (cell copies) eventually differentiate, A third T-cell subset, regulatory T cells (formerly T suppres-
becoming plasma cells that secrete huge numbers of antibody sor cells), has recently been described and is currently the focus of
molecules. Before the naive B cell can multiply, however, it a great deal of research. Regulatory T cells are similar to the other
generally needs to be activated by another type of lympho- T cells in that they have a TCR, but their role is entirely differ-
cyte—a TH cell. The TH cell must also recognize the antigen, ent. Instead of fostering a response, they help prevent the immune
confirming that it is indeed dangerous. system from mounting a response against “self” molecules; fail-
ure to do this results in autoimmune diseases—a topic discussed
in chapter 17. The description of T cells in this chapter focuses
Overview of Cell-Mediated Immunity exclusively on the cytotoxic and helper subsets.
Cell-mediated immunity involves T lymphocytes, or T cells (see Like the B cells, naive helper T cells and naive cytotoxic
figure 15.1). Their name reflects the fact that they mature in the T cells that bind antigen must be activated before they can mul-
thymus. Two subsets of T cells help eliminate antigen—cytotoxic tiply. Again, this process helps confirm that the antigen signi-
 Part III Microorganisms and Humans 357

fies danger. Dendritic cells are responsible for T-cell activation;

B cell T cell
recall that these cells are a part of innate immunity. Once acti-
vated, a T cell proliferates and then differentiates to form effector
T cells, armed to perform distinct protective roles. For simplicity
and clarity, we will refer to effector helper T cells as TH cells, and
effector cytotoxic cells as TC cells. Like B cells, both types of
Antigen-binding
Plasma Antigen-binding site T cells can form memory cells that react quickly if the same anti-
membrane site gen is encountered later. dendritic cell, p. 340
CD marker
TC cells respond to intracellular antigens (antigens within a
host cell). When they find such an antigen, they induce the “self”
Antigen
cell that harbors it to undergo apoptosis. Virally infected cells
Antigen
provide a good example of the effectiveness of this strategy. By
B-cell T-cell receptor
receptor (BCR) (TCR)
inducing these cells to undergo apoptosis, the immune system
destroys cells that would otherwise produce more viral particles.
Sacrificing the cells also releases unassembled viral components,
provoking a humoral response as well. apoptosis, p. 350
FIGURE 15.2 Antigen Receptors on Lymphocytes B cells and TH cells help orchestrate the various responses of humoral and
T cells have surface receptors that allow them to recognize specific
cell-mediated immunity. They activate B cells and macrophages,
antigens. The CD marker identifies the type of T cell.
and produce cytokines that direct and support T cells.
? How is a B-cell receptor similar to an antibody?
MicroAssessment 15.1
B cells are programmed to eliminate extracellular antigens. Cytotoxic
T cells are programmed to destroy infected “self” cells as a means of
eliminating intracellular antigens. Helper T cells are programmed to
orchestrate the adaptive immune response.
Tonsil 1. How is the strategy of the cell-mediated response different from
that of the humoral response?
2. Why is it important that B cells and T cells be activated before
they can mount a response against antigen?
3. How would you expect a TC cell to respond if it encountered
Thymus
a TH cell that was infected with a virus? +
SALT
(skin-associated
lymphoid tissue)

Spleen
15.2 ■ Anatomy of the
MALT (mucosa-
Lymphatic System
associated
lymphoid tissue), Learning Outcome
including 2. Compare and contrast the roles of lymphatic vessels, the various
Peyer’s patches
Bone secondary lymphoid organs, and primary lymphoid organs.
marrow
The lymphatic system is a collection of tissues and organs that
bring the population of B cells and T cells into contact with anti-
gens (figure 15.3). This is important because each lymphocyte
recognizes only one or a few different antigens. In order
Lymph node for the body to mount an effective response, the appro-
priate lymphocyte must encounter the given antigen.
Follicles tissues, p. 69 organs, p. 69
(where naive
B cells gather)
Paracortex Artery
Lymphatic (where naive Vein
vessels T cells gather)

Efferent (outgoing)
lymphatic vessel
Afferent
(incoming) FIGURE 15.3 Anatomy of the Lymphatic
lymphatic System Lymph flows through a system of lymphatic
vessels
vessels, passing through lymph nodes and lymphoid
tissues.

? What is the function of the lymph nodes?

358 Chapter 15 The Adaptive Immune Response

Lymphatic Vessels Some secondary lymphoid organs are less organized in

structure than the lymph nodes and spleen, but their purpose
Flow within the lymphatic system occurs via the lymphatic
is the same—to capture antigens and bring them into contact
vessels, or lymphatics. These vessels carry lymph, a fluid that
with lymphocytes. Among the most important are the Peyer’s
forms as a result of the body’s circulatory system (see figure 27.1).
patches, tissues in the intestinal walls that inspect samples
As blood travels from the heart and lungs through the capil-
of intestinal contents. Specialized intestinal epithelial cells
laries in the tissues, much of the fluid portion filters out, sup-
called M cells transfer material from the intestinal lumen
plying tissues with the O2 and nutrients carried by the blood
to the Peyer’s patches (figure  15.5). Dendritic cells in and
(figure  15.4). Most of that fluid then reenters the capillaries as
near the Peyer’s patches can also reach through the epithelial
they return to the heart and lungs, but some enters the lymphatics
layer and grab material in the intestine to present it to lym-
instead. The lymph—which also contains white blood cells and
phocytes. Peyer’s patches are part of a network of lymphoid
antigens that have entered the tissues—travels via the lymphatics
tissues called mucosa-associated lymphoid tissue (MALT).
to the lymph nodes, where proteins, cells, and other materials are
These play a critical role in mucosal immunity, the immune
removed. The lymph then empties back into the blood circulatory
response that prevents microbes from invading the body via
system at a large vein behind the left collarbone. Note that the
the mucous membranes. Lymphoid tissues under the skin are
inflammatory response causes more fluid to enter the tissues at
called skin-associated lymphoid tissue (SALT). lumen p. 69
the site of inflammation; this causes a corresponding increase in
the antigen-containing fluids that enter lymphatic vessels.
Primary Lymphoid Organs
Secondary Lymphoid Organs Primary lymphoid organs include the bone marrow and thymus.
The bone marrow is where hematopoietic stem cells reside; recall
Secondary lymphoid organs are the sites where lymphocytes
that these give rise to all blood cells, including lymphocytes (see
gather to contact the various antigens. Examples include the
lymph nodes, spleen, tonsils, adenoids, and appendix (see figure
15.3). They are situated at strategic positions in the body so that
immune responses can be initiated almost anywhere. For instance, Antigens
lymph nodes capture materials from the lymphatics, and the
spleen collects materials from blood.
Lumen of Absorptive
The secondary lymphoid organs are like busy, highly orga- epithelial cell
intestine M cell
nized lymphoid coffee shops where many cellular meetings take
place. The anatomy of these organs provides a structured center
for various cells of the immune system to interact and transfer
cytokines. No other places in the body do this, so these organs are
the only sites where adaptive immune responses can be initiated.

Dendritic
Lymphatic
cell
vessel
Excess tissue fluid
becomes lymph Macrophage

Tissue
fluid

Peyer’s
patch
Blood
flow Area where
Area where B cells gather
T cells gather
Capillary
Lymphatic vessels
that drain to local
Filtration lymph nodes
Absorption
Venule Arteriole
FIGURE 15.5 Peyer’s Patch M cells transfer samples of intestinal
FIGURE 15.4 Formation of Lymph contents to lymphocytes that reside in Peyer’s patches.

? What material is typically contained in lymph? ? What is mucosal immunity?

 Part III Microorganisms and Humans 359

figure  14.5). B cells and T cells all originate in the bone marrow
but only B cells mature there; immature T cells migrate to the thy-
mus and mature there. Once mature, the lymphocytes gather in the
secondary lymphoid organs just described, waiting to encounter Antibodies
antigen. hematopoietic stem cells, p. 338

MicroAssessment 15.2
Epitopes
Lymphatic vessels carry fluid collected from tissues to the lymph (antigenic
nodes. Secondary lymphoid organs are where lymphocytes gather to determinants)
encounter antigens. Hematopoietic stem cells destined to become
B cells and T cells mature in the primary lymphoid organs.
4. How is lymph formed?
5. What are Peyer’s patches? Bacterial cell
6. How would mucosal immunity prevent diarrheal disease? +

15.3 ■ The Nature of Antigens Epitopes

(antigenic
determinants)
Learning Outcome
3. Define the terms antigen, immunogen, T-dependent antigen,
T-independent antigen, antigenic, and epitope.
FIGURE 15.6 Antibodies Binding to Epitopes

The term antigen was initially coined in reference to compounds ? Explain why a pathogen would never have only a single epitope.
that induce antibody production; it is derived from the descrip-
tive expression antibody generator. Today, the term is used
more broadly to describe any molecule that reacts specifically
with an antibody, a B-cell receptor, or a T-cell receptor; it does known as epitopes, or antigenic determinants (figure 15.6). Some
not necessarily imply that the molecule can induce an immune epitopes are stretches of 10 or so amino acids, whereas others are
response. When referring specifically to an antigen that elicits an three-dimensional shapes such as a region that sticks out in a mol-
immune response in a given situation, the more restrictive term ecule. A bacterial cell usually has a diverse assortment of macro-
immunogen may be used. The distinction between the terms anti- molecules on its surface, each with a number of distinct epitopes,
gen and immunogen helps clarify discussions in which a normal so the entire cell has a multitude of different epitopes.
protein from host A elicits an immune response when transplanted
into host B; the protein is an antigen because it can react with an
antibody or lymphocyte, but it is an immunogen only for host B, MicroAssessment 15.3
not for host A.
The adaptive immune response is directed against epitopes on
Antigens include an enormous variety of materials, antigens.
from invading microbes and their products to plant pollens,
7. How is an epitope different from an antigen?
but they fall into two general categories. Most antigens are
T-dependent antigens, meaning that the responding B cell 8. Would a denatured antigen be expected to have the same
epitopes as its native (undenatured) counterpart? +
requires assistance from a TH cell in order to become acti-
vated. T-dependent antigens characteristically have a protein
component. In contrast, T-independent antigens can activate
B cells without TH cell help. They include lipopolysaccharide
(LPS) and molecules with repeating subunits, such as some 15.4 ■ The Nature of Antibodies
carbohydrates.
Various antigens differ in their effectiveness in stimulating Learning Outcomes
an immune response. Proteins generally induce a strong response, 4. Diagram an antibody, labeling the various functional regions.
whereas lipids often do not. The terms antigenic and immuno- 5. Describe six protective outcomes of antibody-antigen binding.
genic are used interchangeably to describe the ability of an antigen 6. Compare and contrast the five classes of immunoglobulins.
to elicit an immune response. Small molecules are usually not
antigenic. The structure and characteristics of antibody molecules are criti-
Although antigens are generally large molecules, the adap- cal for their functional properties. Recognizing these features will
tive immune system recognizes discrete regions of the molecule help you understand their essential roles in the host defenses.
360 Chapter 15 The Adaptive Immune Response
Antigen-
binding site

Variable
region
Fab region
Light
chain

Fc region Constant
region
Heavy
chain

(a) (b) (c)

FIGURE 15.7 Structure of an Antibody Molecule (a) Simplified Y-shaped structure; the arms of the Y make up the Fab regions, and the
stem is the Fc region. (b) The molecule is made up of two identical heavy chains and two identical light chains. Disulfide bonds link the chains.
(c) The amino acid sequence of the variable region accounts for the antigen-binding specificity; the amino acid sequence of the constant region
determines the class of the molecule.

? How does the function of the constant region differ from that of the variable region?

Structure and Properties region allows other components of the immune system to recog-
of Antibodies nize the otherwise diverse antibody molecules.
There are five general types of constant regions, and these cor-
Antibodies, also called immunoglobulins, are Y-shaped proteins that respond to the major classes of immunoglobulin (Ig) molecules—
have two general parts—the arms and the stem (figure  15.7a). The IgM, IgG, IgA, IgD, and IgE. Each class has distinct functions
two identical arms, called the Fab regions, bind antigen. The stem is and properties that will be described later, after we consider some
the Fc region. These names were assigned following early studies that general properties of antibodies.
showed that enzymatic digestion of antibodies yielded two types of
fragments—fragments that were antigen-binding (Fab) and fragments
that could be crystallized (Fc). Protective Outcomes
All antibodies have the same basic Y-shaped structure, called
an antibody monomer. It consists of two copies of a high-molecular-
of Antibody-Antigen Binding
weight polypeptide chain, called the heavy chain, and two copies of The protective outcomes of antibody-antigen binding (figure 15.8)
a lower-molecular-weight polypeptide chain, called the light chain depend partly on the antibody class, and include:
(figure 15.7b). The amino acids in the chains fold into characteristic ■ Neutralization. Toxins and viruses must bind specific mol-
domains, referred to as immunoglobulin domains; the light chains ecules on a cell surface before they can damage that cell. A
have two domains each, and most heavy chains have four. Each toxin or virus coated with antibodies cannot attach to cells
light chain is linked to a heavy chain by a disulfide bond. The fork and is said to be neutralized.
of the Y is a flexible stretch called the hinge region, and one or
■ Opsonization. Phagocytic cells have receptors for the Fc
more disulfide bonds link the two heavy chains there. domain, p. 28
disulfide bond, p. 27
region of IgG molecules, making it easier for the phagocyte
When the amino acid sequences of antibody molecules that to engulf antibody-coated antigens. Recall from chapter 14
bind to different epitopes are compared, there is tremendous varia- that the complement protein C3b opsonizes antigens; IgG
tion in the portion referred to as the variable region. The other molecules have a similar effect. opsonization by C3b, p. 345
portion is known as the constant region. ■ Complement system activation. Antigen-antibody com-
plexes (commonly called immune complexes) can trigger
Variable Region the classical pathway of complement system activation.
The variable region is the portion at the ends of the Fab regions; When multiple molecules of certain antibody classes are
it accounts for the antigen-binding specificity (figure  15.7c). Part bound to a cell surface, a specific complement system
of this region is the antigen-binding site, the portion that attaches protein attaches to their Fc regions, initiating the cascade.
to a specific epitope. The fit needs to be very precise, because the Recall that activation of the complement system results in
interaction depends on numerous non-covalent bonds to keep the production of the opsonin C3b, initiation of an inflammatory
molecules together. Nevertheless, the antigen-antibody interaction response, and formation of membrane attack complexes.
is reversible, and the molecules can separate, leaving both antigen complement system, p. 344

and antibody unchanged. ■ Immobilization and prevention of adherence. Binding of

antibodies to flagella interferes with a microbe’s ability to
Constant Region move; binding to pili prevents it from attaching to surfaces.
The constant region includes the entire Fc region, as well as part These capabilities are often necessary for a pathogen to infect a
of the two Fab regions (figure 15.7c). The consistent nature of this host, so antibodies that bind to flagella or pili prevent infection.
 Part III Microorganisms and Humans 361

Opsonization

Bacterium

Neutralization Complement System Activation

Phagocyte
Virus Complement
system protein
Toxin Bacterium

Opsonization by C3b
Inflammatory response
Lysis of foreign cells

Antibody-Dependent Cellular Immobilization and Prevention

Cytotoxicity (ADCC) Natural of Adherence
killer cell

Cross-Linking Bacterium
Infected Bacterium
“self” cell Kills cell
Flagellum

FIGURE 15.8 Protective Outcomes of Antibody-Antigen Binding

? Which pathway of complement activation is depicted in this figure?

■ Cross-linking. The two arms of an antibody can bind sepa- IgM is a pentamer. Its large size prevents it from crossing
rate but identical antigen molecules, linking them. The overall from the bloodstream into tissues, so its primary role is to control
effect is that large antigen-antibody complexes form, creating bloodstream infections. The five monomeric subunits give IgM
big “mouthfuls” of antigens for phagocytic cells to engulf. a total of 10 antigen-binding sites, so it cross-links antigens very
■ Antibody-dependent cellular cytotoxicity (ADCC). When effectively. It is the most efficient class in triggering the classical
multiple IgG molecules bind to a virally infected cell or a pathway of complement system activation.
tumor cell, that cell becomes a target for destruction by natural A fetus is normally sterile until the birth membrane is rup-
killer (NK) cells. The NK cell attaches to the Fc regions of IgG tured, so IgM generally begins being made about the time of birth.
and once attached, kills the target cell by delivering compounds However, a fetus infected in utero can make IgM antibodies.
directly to it. natural killer cells, pp. 340, 374
IgG
IgG accounts for about 80% to 85% of the total serum immu-
noglobulin (serum is the liquid portion of blood). It circulates in
Immunoglobulin Classes
the blood but exits the vessels to enter the tissues as well.
All five major classes of immunoglobulins have the same basic IgG provides the longest-term protection of any antibody
monomeric structure, but each class has a different constant class; its half-life is 21 days, meaning that a given number of IgG
portion of the heavy chain. Some of the immunoglobulins form molecules will be reduced by about 50% after 21 days. In addi-
multimers of the basic monomeric structure. Characteristics of the tion, IgG is generally the first and most abundant circulating class
various immunoglobulin classes are summarized in table 15.1. produced during the secondary response. The basis for this will be
discussed later in the chapter. IgG provides protection by neutral-
IgM ization, opsonization, complement activation, immobilization and
IgM accounts for 5% to 13% of the circulating antibodies and prevention of adherence, cross-linking, and ADCC.
is the first class produced during the primary response to an An important characteristic of IgG is that it is transported
antigen. It is the principal class produced in response to some across the placenta into the fetus’s bloodstream, so it protects the
T-independent antigens. developing fetus against infections. Women who are not already
362 Chapter 15 The Adaptive Immune Response

TABLE 15.1 Characteristics of the Main Classes of Antibodies

Class and Percent of
Molecular Total Serum
Weight Immunoglobulin
(daltons) Structure (half-life in serum) Properties and Functions

IgM 970,000 5–13% (10 days) First antibody class produced during the primary response. Principle class
produced in response to T-independent antigens. Provides direct protection
by neutralizing viruses and toxins, immobilizing motile organisms,
preventing microbes from adhering to cell surfaces, and cross-linking
antigens. Binding of IgM to antigen leads to activation of the complement
system (classical pathway).

IgG 146,000 80–85% (21 days) Most abundant class in the blood and tissue fluids. Provides longest-term
protection because of its long half-life. Transported across the placenta,
providing protection to a developing fetus; long half-life extends the
protection through the first several months after birth. Provides direct
protection by neutralizing viruses and toxins, immobilizing motile
organisms, preventing microbes from adhering to cell surfaces, and cross-
linking antigens. Binding of IgG to antigen facilitates phagocytosis, leads
to activation of the complement system (classical pathway), and allows
antibody-dependent cellular cytotoxicity.
IgA monomer 10–13% (6 days) Most abundant class produced, but the majority of it is secreted into
160,000; mucus, tears, and saliva, providing mucosal immunity. Also found in
secretory IgA breast milk, protecting the intestinal tract of breast-fed infants. Protects
390,000 mucous membranes by neutralizing viruses and toxins, immobilizing motile
organisms, and preventing attachment of microbes to cell surfaces.

IgD 184,000 <1% (3 days) Involved in the development and maturation of the antibody response. Its
functions in blood are not well understood.

IgE 188,000 <0.01% (2 days) Binds via the Fc region to mast cells and basophils. This bound IgE allows
those cells to detect parasites and other antigens and respond by releasing
their granule contents. Involved in many allergic reactions.

immune to a given pathogen lack IgG against that microbe, so

they are warned to take extra precautions during pregnancy to 100
avoid pathogens that can infect and damage a fetus. For exam-
ple, pregnant women are advised not to eat raw meat or become
Percent of normal average

first-time cat owners to avoid a primary infection by Toxoplasma

gondii, a parasite found in steak tartare and other raw meat as well
adult level of IgG

as the feces of infected cats. Toxoplasma gondii, p. 709

Maternal IgG not only protects the developing fetus, but also
Total IgG
the newborn. The maternal antibodies present at birth gradually
degrade over a period of about 6 months, but during this time the
infant begins producing protective antibodies (figure 15.9).
IgG is also in colostrum, the first breast milk produced after
giving birth. The newborn’s intestinal tract absorbs this antibody. Infant IgG Maternal IgG

IgA 0
The monomeric form of IgA accounts for about 10% to 13% of 4 6 8 2 4 6 8
Before birth Birth Infant age
antibodies in the serum. Most IgA, however, is the secreted form,
a dimer called secretory IgA (sIgA). In fact, IgA is the most Months
abundant immunoglobulin class produced. The secreted form is
important in mucosal immunity and is found on the mucous mem- FIGURE 15.9 Immunoglobulin G Levels in the Fetus and
branes that line the gastrointestinal, genitourinary, and respiratory Infant As the fetus develops, maternal IgG is transported across the
tracts. It is also in secretions such as saliva, tears, and breast milk. placenta to provide protection. After birth, the infant begins produc-
ing antibodies.
Secretory IgA in breast milk protects breast-fed infants against
intestinal pathogens. mucosal immunity, p. 358 ? Why does the level of maternal antibodies decrease over time?
 Part III Microorganisms and Humans 363

Protection by secretory IgA is primarily due to the direct antigen, and only those cells begin multiplying. This generates a
effect of its binding. These include neutralizing toxins and viruses population of clones—copies of the specific B cells capable of
and interfering with the attachment of microbes to host cells. making the appropriate antibodies.
IgA is produced by the plasma cells that reside in the mucosa- Clonal selection is a critical theme in the adaptive immune
associated lymphoid tissues (MALT). Recall that plasma cells response, pertaining to both B cells and T cells. As lymphocytes
are the antibody-secreting form of B cells. As IgA is transported mature in the primary lymphoid organs, a population of cells able
across the mucosa (mucous membranes), a polypeptide called the to recognize a functionally limitless variety of antigens is gener-
secretory component is added. This attaches the antibody to the ated; each individual cell, however, recognizes and responds to
layer of mucus that coats the mucosal surface and protects it from only one epitope. Thus, if a person’s immune system can make
destruction by enzymes there. MALT, p. 358 antibodies to billions of different epitopes, that person must have
billions of different B cells, each interacting with a single epitope.
IgD The process of generating the diversity in antigen recognition is
IgD accounts for less than 1% of all serum immunoglobulins. It random and does not require previous exposure to an epitope; the
is involved with the development and maturation of the antibody mechanisms will be described later.
response, but its functions in blood have not been clearly defined. Each lymphocyte residing in the secondary lymphoid organs
is waiting for the “antigen of its dreams”—an antigen that has an
IgE epitope to which that particular lymphocyte is programmed to
IgE is barely detectable in blood, because most is tightly bound respond. When an antigen enters a lymphoid organ, only those
via the Fc region to basophils and mast cells, rather than being rare lymphocytes that recognize it can respond; the specificity
free in the circulation. The bound IgE molecules allow these of the antigen receptor they carry on their surface (B-cell recep-
cells to detect and respond to antigens. For example, when anti- tor or T-cell receptor) determines this recognition. Lymphocytes
gen binds to two adjacent IgE molecules carried by a mast cell, that do not recognize the antigen remain inactive. Recall that in
the cell releases histamine and other inflammatory mediators. most cases, lymphocytes require additional signals—confirmation
IgE-mediated responses seem to be important in eliminating from another cell type—in order to multiply. This helps prevent
parasites, particularly helminths. inflammatory mediators, p.  348
the immune system from mounting a response against “self” mol-
helminths, p. 295
ecules by mistake.
Unfortunately for allergy sufferers, basophils and mast cells Some progeny of the lymphocytes that encountered their
also release their chemicals when IgE binds to normally harmless “dream antigen” leave the secondary lymphoid organs and
materials such as foods, dusts, and pollens, leading to immedi- migrate to the tissues where they continue responding for as long
ate reactions such as coughing, sneezing, and tissue swelling. In as the antigen is present. Without continuous stimulation by anti-
some cases these allergic, or hypersensitivity, reactions can be gen, these cells will undergo apoptosis. apoptosis, p. 350
life-threatening. hypersensitivity reactions, p. 401 The activities of individual lymphocytes change as they
encounter antigen. As a means of clarifying discussions of lym-
phocyte characteristics, descriptive terms are sometimes used:
MicroAssessment 15.4
Antibody-antigen binding results in neutralization, immobilization, ■ Immature lymphocytes. These have not fully developed
and prevention of adherence, cross-linking, opsonization, complement their antigen-specific receptors.
activation, and antibody-dependent cytotoxicity. Immunoglobulin ■ Naive lymphocytes. These have antigen receptors, but
classes include IgM, IgG, IgA, IgD, and IgE. have not yet encountered the antigen to which they are pro-
9. Why is IgM particularly effective at cross-linking antigens? grammed to respond.
10. Which maternal antibody classes protect a breast-fed newborn? ■ Activated lymphocytes. These are able to proliferate; they
11. In opsonization with IgG, why is it important that phagocytes have bound antigen via their antigen receptor and have
recognize the antibodies only after they bind antigen? + received the required accessory signals from another cell,
confirming that the antigen merits a response.
■ Effector lymphocytes. These are descendants of activated
15.5 ■ Clonal Selection and lymphocytes, armed with the ability to produce specific cyto-
Expansion of Lymphocytes kines or other protective substances. Plasma cells are effector
B cells, TC cells are effector cytotoxic T cells, and TH cells are
Learning Outcome effector helper T cells.
7. Outline the process of clonal selection and expansion. ■ Memory lymphocytes are long-lived descendants of activated
lymphocytes; they can quickly become activated when an anti-
Early on, immunologists recognized that the immune system is gen is encountered again. Memory lymphocytes are respon-
capable of making a seemingly infinite array of antibody speci- sible for the speed and effectiveness of the secondary response.
ficities. The clonal selection theory describes how this occurs
MicroByte
(figure 15.10). Each B cell is programmed to make only a single The body is thought to have about 1 billion B cells, and only one or
specificity of antibody. When antigen is introduced, only the B a few will recognize a given epitope.
cells capable of making the appropriate antibody can bind to the
364 Chapter 15 The Adaptive Immune Response

FIGURE 15.10 Clonal Selection and Expansion During

the Antibody Response
? What is meant by clonal selection?

Development
Hematopoietic stem cell
Immature B cells: As
these develop, a
functionally limitless
assortment of B-cell
receptors is randomly Antigen X
generated.

Naive B cells: Each

cell is programmed to
recognize a specific
epitope on an antigen;
B-cell receptors guide B cell W B cell X B cell Y B cell Z
that recognition. recognizing antigen X

Activation Selected B cell receives confirmation from a specific

TH cell that a response is warranted (not shown
Activated B cells: here; process is illustrated in figure 15.11)
These cells can
proliferate because
their B-cell receptors
are bound to antigen X
and the cells have
received required
signals from TH cells.

Proliferation and
differentiation

Plasma cells
(effector B cells):
These descendants of
activated B cells
secrete large quantities
of antibody molecules
that bind to antigen X.

Memory B cells:
These long-lived
descendants of
activated B cells
recognize antigen X
when it is encountered
again.

Effector action
Antibodies:
These neutralize the
invader and tag it for
destruction.
 Part III Microorganisms and Humans 365

MicroAssessment 15.5 B-Cell Activation

In response to an antigen, only those lymphocytes that recognize Naive B cells gather in the secondary lymphoid organs to encoun-
the antigen proliferate. Depending on their developmental stage, ter antigens. When a B cell’s antigen receptor (B-cell receptor)
lymphocytes may be referred to as immature, naive, activated, binds to a T-dependent antigen, the B cell takes the antigen in by
effector, or memory cells. endocytosis, enclosing it within an endosome. There, the antigen
12. Describe the clonal selection theory. is degraded into peptide fragments that are then delivered to pro-
13. How does a naive lymphocyte differ from an activated one? tein structures called MHC class II molecules. These move to
14. If the heavy chain of an antibody is approximately 450 amino the B-cell surface, where they “present” pieces of the antigen for
acids long, how much DNA would be required to encode 10 9 inspection by TH cells—a process called antigen presentation
separate heavy chain genes? + (figure 15.11). endocytosis, p. 72
TH cells, which also gather in the secondary lymphoid organs,
scan the naive B cells there to determine if any have encountered
15.6 ■ B Lymphocytes and an antigen they recognize. If a TH cell’s antigen receptor (T-cell
receptor) binds one of the peptide fragments being presented by a
the Antibody Response B cell, then that T cell activates the B cell. It does this by deliver-
ing cytokines to the B cell, initiating the process of clonal expan-
Learning Outcomes
sion of that particular B cell.
8. Describe the role of TH cells in B-cell activation.
If no TH cells recognize the peptides presented by a B cell,
9. Compare and contrast the primary and the secondary responses. that B cell may become anergic (unresponsive to future exposure
10. Compare and contrast the response to T-dependent antigens and to the antigen). This results in tolerance to that antigen, a mechan-
T-independent antigens. ism the adaptive immune system uses to avoid responses against
“self” and other harmless antigens. tolerance, p. 355
Recall that most antigens are T-dependent antigens, mean-
ing the B cells that recognize them require help from TH cells;
the response to these antigens will be the primary focus of this
section. The B-cell response to T-independent antigens will be
Characteristics of the
covered at the end of this section. Later in the chapter, we will Primary Response
explain how naive helper T cells become activated to develop their In the first (primary) exposure to an antigen, it takes about 10
effector functions. to 14 days for a substantial amount of antibodies to accumulate

1 B-cell receptor 2 3

Antigen Endosome

B-cell receptor binds B cell internalizes antigen. B cell degrades antigen into
to antigen. peptide fragments.

5a T-cell receptor

4
Antigen MHC class II
fragment molecule TH cell recognizes
antigen fragment FIGURE 15.11 B-Cell
Microbial and activates B cell. Activation The B cell
antigen processes the antigen and
presented. Cytokine delivery presents it to TH cells. If a TH
cell recognizes the antigen,
Harmless 5b it activates the B cell, allow-
antigen ing it to undergo clonal
presented. No TH cell recognizes expansion.
Peptide fragments are antigen fragment;
presented on MHC class II B cell becomes
molecules.
? What happens if no
anergic. TH cells recognize the
antigen presented by
the B cell?
366 Chapter 15 The Adaptive Immune Response

antigen is present. The result is a slow but steady increase in the

titer (concentration) of antibody molecules.
Primary Secondary
response response Over time, the proliferating B cells undergo changes that
Concentration of antibody

improve the immune response. These include:

IgG ■ Affinity maturation. This is a form of natural selection
among proliferating B cells (figure  15.14). As activated
B cells multiply, spontaneous mutations commonly occur
in certain regions of the antibody genes. Some of these
result in slight changes in the antigen-binding site of the
IgG antibody (and therefore the B-cell receptor). B cells that
IgM bind antigen for the longest duration are most likely to
Days Months Days Months proliferate.
Ag Ag ■ Class switching. All B cells are initially programmed to
Time after antigen (Ag) injection differentiate into plasma cells that secrete IgM. Cytokines
produced by TH cells, however, induce some activated
FIGURE 15.12 The Primary and Secondary Responses to B cells to switch that genetic program, causing them to
Antigen The first exposure to antigen elicits relatively low amounts differentiate into plasma cells that secrete other antibody
of first IgM, followed by IgG in the blood. The second exposure,
which characterizes the memory of the adaptive immune system,
classes. B cells in the lymph nodes most commonly switch
elicits rapid production of relatively large quantities of IgG. to IgG production (figure  15.15). B cells in the mucosa-
associated lymphoid tissues generally switch to IgA produc-
? How would this graph be different if it illustrated antibody levels
tion, providing mucosal immunity.
on a mucosal surface?
After class switching, some B cells become memory
B cells. These persist in the body for years and are present in
(figure  15.12). During this delay, the person might experi- numbers sufficient to give a prompt secondary response if the
ence signs and symptoms of an infection, which could be life- same antigen is encountered again later.
threatening. The immune system, however, is actively responding. The antibody response begins to wane as the accumulating
Naive B cells that bind the antigen present the peptide fragments antibodies clear the antigen. Progressively fewer molecules of
to TH cells. Once activated, those B cells multiply, generating a antigen remain to stimulate the lymphocytes, and, as a result,
population of cells that recognize the antigen. As some of the the activated lymphocytes undergo apoptosis. Memory B cells,
activated B cells continue dividing, others differentiate to form however, are long-lived even in the absence of antigen.
antibody-secreting plasma cells (figure 15.13).
MicroByte
Each plasma cell generally undergoes apoptosis after several A plasma cell secretes thousands of identical antibody molecules
days, but activated B cells continue multiplying and differentiat- per second.
ing, generating increasing numbers of plasma cells as long as

Nucleus Rough endoplasmic reticulum

(a) 10 µm (b) 10 µm (c) 10 µm

FIGURE 15.13 Lymphocytes and Plasma Cells (a) Light micrograph of a T lymphocyte. The morphology is the same as that of a B lymphocyte.
(b) Scanning electron micrograph of a B lymphocyte. (c) Plasma cell, an effector B cell, which secretes antibody molecules. Note the extensive rough endo-
plasmic reticulum, the site of protein synthesis.

? Would a single plasma cell produce antibody molecules of the same specificity or different specificities? Explain your answer.
 Part III Microorganisms and Humans 367

Characteristics of
++
the Secondary Response
The secondary response is significantly faster and more effective
than the primary response. In fact, repeat invaders are generally
eliminated before they cause noticeable harm. This is why a per-
+ ++ son who has recovered from a particular disease generally has
long-lasting immunity to that disease. Vaccination takes advan-
tage of this naturally occurring phenomenon. vaccination, p. 421
Memory B cells are responsible for the efficiency of the sec-
ondary response. For one thing, there are more cells—memory
+++ ++ B cells as well as memory helper T cells—that can respond to a
specific antigen. In addition, the memory B cells are able to scav-
enge even low concentrations of antigen because their receptors
have been fine-tuned through affinity maturation. Likewise, the
antibodies coded for by these cells bind antigen more effectively.
+++ +++ When memory B cells become activated, some quickly dif-
ferentiate to form plasma cells, resulting in the rapid production
of antibodies. Because of class switching, these antibodies are
often IgG or IgA. Other activated cells begin proliferating, once
again undergoing affinity maturation to generate even more effec-
tive antibodies. Subsequent exposures to antigen lead to an even
+++ +++ +++ +++ stronger response.

The Response to
FIGURE 15.14 Affinity Maturation B cells that bind antigen for
the longest duration are the most likely to proliferate. The plus signs T-Independent Antigens
indicate the relative quality of binding of the antibody to the anti- T-independent antigens can activate B cells without the aid of TH
gen; those in green indicate the most “fit” to continue proliferating.
cells. Relatively few antigens are T-independent, but they can be
? What accounts for the change in a B cell’s ability to bind antigen? very important medically.

FIGURE 15.15 Class Switching Naive B cells are pro-

grammed to produce IgM antibodies. Activated B cells
undergo class switching. Class switching does not alter the
variable region and therefore has no effect on the antibody Pr
P ro
og
o g
Programmed for
specificity. The plasma cells descended from B cells in lymph Ig
gM e
IgM expression
nodes most commonly produce IgG after class switching.

? How would this illustration change if it showed

N los
No lloss
ss off DNA Lo
Loss of DNA
class switching by B cells residing in Peyer’s patches?
Variable region IgM IgD IgG IgE IgA

Segment of DN
NA
NA
DNA
B-cell DNA delete
ed
deleted

Pr
P rogrammme for
m
Programmed Pr
P rogra
am
amm for
Programmed
Ig
gM expr
IgM re
es
e
expression Ig
gG ex
IgG xppr
expression

e rregion
Variable egion IgM Ig
g
gD
IgD IgG IgE IgA Variable region IIgG IgE IgA

B-cell DNA B-cell DNA

Plasma cells that develop from this B cell will produce IgM because that Plasma cells that develop from this B cell will produce IgG because
constant region gene segment is first in line. the DNA loss now puts that constant region gene segment first in line.
368 Chapter 15 The Adaptive Immune Response

Polysaccharide
Polys
P l saaccha
ar antigen MicroAssessment 15.6
with
w multiple
multip
m ple repeating
e rep
pea
subunits
sub
buunits In most cases, B cells that bind antigen require accessory signals from
TH cells to become activated. Activated B cells proliferate, ultimately
B-cell becoming either plasma cells that secrete antibody molecules or
receptorss long-lived memory cells. Affinity maturation and class switching
occur in the primary response; these allow a swift and more effective
secondary response. T-independent antigens can stimulate an antibody
response by activating B cells without the aid of TH cells.
15. Describe the significance of class switching.
16. How does the ability to bind antigen increase as B cells multiply?
17. Why should B cells residing in the mucosa-associated lymphoid
tissues produce IgA? +

15.7 ■ T Lymphocytes: Antigen

FIGURE 15.16 T-Independent Antigens Antigens such as some
polysaccharides have multiple repeating epitopes. Because of the Recognition and Response
arrangement of epitopes, clusters of B-cell receptors bind to the anti-
gen simultaneously, leading to B-cell activation without the involve- Learning Outcomes
ment of TH cells.
11. Describe the importance of T-cell receptors and CD markers.
? Why is the response to T-independent antigens important medically? 12. Describe the role of dendritic cells in T-cell activation.
13. Compare and contrast TH and TC cells with respect to antigen
Polysaccharides and other molecules that have numerous identi- recognition and the response to antigen.
cal evenly spaced epitopes are one type of T-independent antigen.
Because of the arrangement of epitopes, clusters of B-cell receptors The role of T cells is very different from that of B cells (see figure
bind the antigen simultaneously, leading to activation of that B cell 15.1). For one thing, T cells never produce antibodies. Instead,
without the involvement of helper T cells (figure  15.16). These effector T cells directly interact with other cells—target cells—to
T-independent antigens are particularly significant because they cause distinct changes in those cells (table 15.2).
are not very immunogenic in young children. This is why children
less than 2 years of age are more susceptible to pathogens such as
Streptococcus pneumoniae and Haemophilus influenzae, which General Characteristics of T Cells
cloak themselves in polysaccharide capsules. Antibodies that bind Like B cells, T cells have multiple copies of a receptor on their
the capsules would be protective if the child’s immune system could surface that recognizes a specific epitope. The T-cell receptor
make them. Vaccines made from purified capsules are available, (TCR) has two polypeptide chains (a set of either alpha and beta
but, likewise, they do not elicit an immune response in young chil- or gamma and delta), each with a variable and constant region.
dren. Fortunately, newer vaccines designed to induce a T-dependent From a structural standpoint, the T-cell receptor can be compared
response have been developed. They will be discussed later, when the to one “arm” of the B-cell receptor (figure 15.17).
role of TH cells in the antibody response is described in more detail. Unlike the B-cell receptor, the T-cell receptor does not
Lipopolysaccharide (LPS), a component of the outer mem- interact with free antigen. Instead, the antigen must be “pre-
brane of Gram-negative bacteria, is another type of T-independent sented” by another host cell, as described in the section on B-cell
antigen. The constant presence of antibodies against LPS is activation (see figure 15.11). The host cell does this by partly
thought to provide an early defense against Gram-negative bacteria degrading (processing) the antigen and then displaying (present-
that breach the body’s barriers. ing) individual peptides of the antigen’s proteins. The peptides

TABLE 15.2 Characteristics of T Cells

Source of
Antigen
T Cell Type/ Antigen Potential Effector Recognized by
CD Marker Recognition Effector Form Target Cells Function Effector Cell

Cytotoxic T cell/CD8 Peptides presented TC cell All nucleated cells Induces target Endogenous
on MHC class I cell to undergo (produced within
molecules apoptosis the target cell)
Helper T cell/CD4 Peptides presented TH cell B cells, Activates target Exogenous
on MHC class II macrophages cell (produced outside
molecules of the target cell)
 Part III Microorganisms and Humans 369

Antigen: Alpha Beta Peptide-binding groove Peptide-binding groove

MHC-binding site chain chain

Variable
region

Constant
region

(a) MHC Class I Molecule (b) MHC Class II Molecule

(a) (b) FIGURE 15.18 MHC Molecules (a) MHC class I molecule; cyto-
plasmic proteins (endogenous antigens) are presented in the groove
FIGURE 15.17 Structure of a T-Cell Receptor (a) Simplified of these molecules. (b) MHC class II molecule; proteins taken in by
structure, showing the antigen:MHC-binding site. (b) The molecule is the cell (exogenous antigens) are presented in the groove of these
made of two different chains, each with a constant region and a vari- molecules.
able region, linked by a disulfide bond.
? Which cell type recognizes peptides presented in MHC class I
? A T-cell receptor is similar to which part of a B-cell receptor? molecules? Which cell type recognizes peptides presented in MHC
class II molecules?
from the antigen are cradled in the groove of proteins—major
histocompatibility complex (MHC) molecules—on the surface
of the presenting cell.
Two types of MHC molecules present antigen—MHC class
I and MHC class II (figure  15.18). Both are shaped somewhat
like an elongated bun and hold the peptide lengthwise, like a
MHC class I
bun holds a hot dog. When a T cell recognizes an antigen, it is CD8 T-cell receptor
molecule
actually recognizing both the peptide and MHC molecule simulta-
neously. In other words, the T-cell receptor recognizes the “whole
sandwich”—the peptide:MHC complex.
MHC class I molecules present endogenous antigens Endogenous
antigen
(antigens made within the cell). MHC class II molecules present
exogenous antigens (antigens taken up by a cell). All nucleated TC cells recognize All nucleated cells present endogenous
cells produce MHC class I molecules, but only specialized cell antigens presented on antigens on MHC class I molecules.
types (dendritic cells, B cells, and macrophages)—collectively MHC class I molecules.
referred to as antigen-presenting cells (APCs)—make MHC (a)
class II molecules (see Perspective 15.1).
Recall from the introductory section that two functionally CD4 T-cell receptor
MHC class II
distinct T-cell populations are involved in eliminating antigen— molecule
cytotoxic T cells and helper T cells. These differ in their roles, Exogenous
and also how they recognize antigen: Cytotoxic T cells recognize antigen
antigen presented on MHC class I molecules, whereas helper
T cells recognize antigen presented on MHC class II molecules.
Because of these recognition characteristics, effector cytotoxic TH cells recognize B cells and macrophages present
T (TC) cells respond to endogenous antigens, whereas effector antigens presented on exogenous antigens on MHC class II
MHC class II molecules. molecules.
helper T (TH) cells respond to exogenous antigens (figure 15.19).
Cytotoxic and helper T cells are identical microscopically,
so scientists distinguish them based on the presence of surface (b)

proteins called cluster of differentiation (CD) markers. Most FIGURE 15.19 Antigen Recognition by Effector T Cells
cytotoxic T cells have the CD8 marker and are frequently referred (a) Cytotoxic T (Tc ) cell. (b) Helper T (TH) cell.
to as CD8 T cells; most helper T cells carry the CD4 marker and ? What is the fate of a cell that presents antigen recognized by a TC
are often called CD4 T cells. Note that CD4 is also a receptor for cell? What is the fate of a cell that presents antigen recognized by
HIV, which explains why the virus infects helper T cells. a TH cell?
370 Chapter 15 The Adaptive Immune Response

PERSPECTIVE 15.1
What Flavors Are Your Major Histocompatibility Complex Molecules?
The major histocompatibility molecules were inherited one set of the three genes from your overwhelm the defenses. The ability to bind
discovered over half a century ago, long before mother and one set from your father; both sets different peptides is not enough, however,
their critical role in adaptive immunity was are expressed. Putting this all together, and because, ideally, a given peptide should be
recognized. During World War II, bombing assuming that you inherited two completely presented in several slightly different orien-
raids caused serious burns in many people, different sets of alleles from your parents, tations so that distinct aspects of the three-
stimulating research into skin transplants to your cells express six different varieties of dimensional structures can be inspected by
replace burned tissue. That research quickly MHC class I molecules—two of the over T-cell receptors. No single variety of MHC
expanded to include transplants of a variety 890 known HLA-A possibilities, two of the molecule can accomplish all of these aims,
of other tissues and organs. Unfortunately, the over 1,400 HLA-B possibilities, and two of which explains the need for their diversity.
transplanted material was generally rejected the over 620 HLA-C possibilities. As you The variety of MHC molecules that a
by the recipient’s immune system because can imagine, the likelihood that anyone you person has on his or her cells affects that indi-
certain molecules on the donor cells dif- encounter in a day will have those same MHC vidual’s adaptive response to certain antigens.
fered from those on the recipient’s cells; the class I molecules is extremely unlikely, unless This is not surprising because MHC molecules
donor tissue was perceived as an “invader” you have an identical twin. differ in the array of peptides they can bind
by the recipient’s immune system. To over- Why is there so much diversity in MHC and manner in which those peptides are held
come this problem, researchers tried to more molecules? The answer lies in the com- in the molecule. Thus, they affect what the
closely match donor and recipient tissues, plex demands of antigen presentation. MHC T cells actually “see.” In fact, the severity of
relying on newly developed tissue-typing class I molecules bind peptides that are only certain diseases has been shown to correlate
tests. The typing tests looked for leukocyte 8 to 10 amino acids in length; MHC class II with the MHC type of the infected indi-
surface molecules called human leukocyte molecules bind peptides that are only 13 to 25 vidual. For example, rheumatic fever, which
antigens (HLAs), which serve as markers for amino acids in length. Somehow, within that can occur as a consequence of Streptococcus
tissue compatibility. Later, researchers deter- constraint, the MHC molecules must bind as pyogenes infection, develops more frequently
mined that HLAs were encoded by a cluster many different peptides as possible in order in individuals with certain MHC types. The
of genes, now called the major histocompat- to ensure that a representative selection from most serious manifestations of schistosomiasis
ibility complex. Unfortunately, the terminol- the proteins within a cell can be presented to have also been shown to correlate with certain
ogy can be confusing because the molecules T cells. The ability to bind a wide variety of MHC types. Epidemics of life-threatening
that transplant biologists refer to as HLAs are peptides is particularly important considering diseases such as plague and smallpox have
called MHC molecules by immunologists. how readily microbes evolve in response to dramatically altered the relative proportion
It is highly unlikely that two random indi- selective pressure. For example, if a single of MHC types in certain populations, killing
viduals will have identical MHC molecules. alteration in a viral protein prevented all MHC those whose MHC types ineffectively present
This is because the genes encoding them are molecules from presenting peptides from that peptides from the causative agent. rheumatic
polygenic, meaning they are encoded by more protein, then a virus with that mutation could fever, p. 489 schistosomiasis, p. 350
than one locus (position on the chromosome)
and each locus is highly polymorphic (multiple
forms). As an analogy, if MHC molecules
FIGURE 1 MHC Set of MHC genes inherited from your mother:
were candy, each cell would be covered with Polymorphisms
pieces of chocolate, taffy, and lollipop. The The order of the HLA-B HLA-C HLA-A
type of chocolate could be dark, white, milk, MHC class I genes
or a number of various flavors; the taffy could on the chromosome
be peppermint, raspberry, or cinnamon, and so is B, C, and A. One of at least 1,400 One of at least 620 One of at least 890
different alleles different alleles different alleles
on. As you might imagine, the number of dif-
ferent combinations are immense.
There are three loci of MHC class I genes, Set of MHC genes inherited from your father:
designated HLA-A, HLA-B, and HLA-C
(figure  1). There are at least 890 alleles HLA-B HLA-C HLA-A
(forms) for HLA-A, 1,400 for HLA-B, and
620 for HLA-C. In addition, the loci are all One of at least 1,400 One of at least 620 One of at least 890
co-dominantly expressed. In other words, you different alleles different alleles different alleles

Many cells involved with the immune response have various

subsets (for example, TH cells include three subsets—TH1, TH2, Activation of T Cells
and TH17). For simplicity, however, we will focus on the general Dendritic cells, the scouts of innate immunity, play a crucial
characteristics of the cell types, rather than the subsets. role in T-cell activation (figure  15.20). Immature dendritic
cells reside in peripheral tissues, particularly under the skin and
MicroByte
mucosa, gathering various materials from those areas. The cells
Of the approximately 1010 T cells in the body, only a few will
recognize a given epitope the first time it is encountered. use both phagocytosis and pinocytosis to take up particulate and
soluble material that could contain foreign proteins. Dendritic
 Part III Microorganisms and Humans 371

Dendritic cells in the tissue collect

particulate and soluble antigen and then
travel to the secondary lymphoid tissues.

Lymphoid organ

MHC class I molecule

Dendritic cells presenting Dendritic cells presenting “self”
Co-stimulatory molecule microbial peptides produce peptides or other harmless
co-stimulatory molecules. material do not produce
MHC class II molecule co-stimulatory molecules.

T-cell receptor T-cell receptor T-cell receptor T-cell receptor

CD4 CD8 CD4 CD8

Naive T cells that recognize antigen presented Naive T cells that recognize antigen presented
by dendritic cells expressing co-stimulatory molecules by dendritic cells not expressing co-stimulatory
can become activated. molecules become anergic.

Activated T cells proliferate and differentiate. Anergic T cells cannot respond and eventually undergo apoptosis.

FIGURE 15.20 T-Cell Activation

? What causes a dendritic cell to produce co-stimulatory molecules?

cells located just below the mucosal barriers are even able to send undergo apoptosis. Recall that inducing anergy is one mechanism
tentacle-like extensions between the epithelial cells of the barri- the adaptive immune response uses to eliminate lymphocytes that
ers. Using this action, the dendritic cells gather material from the recognize “self” proteins, a critical aspect of self-tolerance.
respiratory tract and the lumen of the intestine. Dendritic cells are able to “cross present” antigens, meaning they
Dendritic cells have TLRs (toll-like receptors) and other can present peptides on both types of MHC molecules—class I
receptors that allow them to recognize pathogens. If pathogens are and class II—regardless of the origin. This allows them to pres-
detected, the dendritic cell takes up even more material, and then ent antigen to, and therefore activate, cytotoxic T cells as well as
travels to the secondary lymphoid organs where it will encounter helper T cells.
naive T cells. Dendritic cells near the end of their life span do this Once a T cell is activated, it undergoes clonal selection and
as well. TLRs, p. 342 expansion as described previously, eventually forming effector
En route to the secondary lymphoid organs, the dendritic cells and memory cells. These can leave the secondary lymphoid
cells mature into a form that presents antigen to naive T cells. organs and circulate in the bloodstream. They can also enter tis-
Dendritic cells that detected pathogens produce surface proteins sues, particularly at sites of infection.
called co-stimulatory molecules. These molecules function as Macrophages and B cells also present antigens they have
“emergency lights” that interact with the T cell, communicating gathered on MHC class II molecules and can produce co-
that the material being presented indicates “danger.” Naive T cells stimulatory molecules. Based on this information, it seems they
that recognize antigen presented by dendritic cells displaying co- should also be able to activate naive T cells, but studies sug-
stimulatory molecules can become activated. In contrast, naive T gest they do not contact naive T cells in vivo. They do encounter
cells that recognize antigen presented by a dendritic cell not dis- memory T cells, however, so they probably activate these cells
playing co-stimulatory molecules become anergic and eventually during the secondary response.
372 Chapter 15 The Adaptive Immune Response

Effector Functions of TC (CD8) Cells lysis minimizes the number of intracellular microbes that might
spill into the surrounding area and infect other cells. Most microbes
TC cells induce apoptosis in infected “self” cells. They also
remain in cell remnants until they are ingested by macrophages. The
destroy cancerous “self” cells.
TC cell survives and can go on to kill other targets.
How do TC cells distinguish infected or cancerous cells from
In addition to inducing apoptosis in the target cell, the TC cell
their normal counterparts? The answer lies in the significance of
will also produce various cytokines that strengthen the “security sys-
antigen presentation on MHC class I molecules (figure 15.21). All
tem.” One cytokine increases antigen processing and presentation in
nucleated cells routinely degrade a portion of the proteins they pro-
nearby cells, making it easier for TC cells to find other infected cells.
duce (endogenous proteins). They then load the resulting peptides
Another cytokine activates local macrophages whose TLRs have
into the groove of MHC class I molecules and deliver them to the
been triggered. Note that a more efficient mechanism of macrophage
surface for inspection by circulating TC cells (see figure 15.19a). If a
activation involves TH cells and will be discussed shortly.
“self” cell is producing only normal proteins, then the peptides being
presented should not be recognized by any circulating TC cells. If the
“self” cell harbors a replicating virus or microorganism, however, Effector Functions of TH (CD4) Cells
then some of the peptides presented on MHC class I molecules will
be recognized by circulating TC cells. This makes the presenting cell TH cells orchestrate the immune response. They activate B cells
a target for the lethal effector functions of TC cells. The same thing and macrophages and direct the activities of B cells, macrophages,
can occur if the “self” cell is producing abnormal proteins that char- and T cells.
acterize cancerous cells. TH cells recognize antigen presented on MHC class II mol-
When a TC cell encounters a cell presenting a peptide it rec- ecules. Recall that these are found only on antigen-presenting cells
ognizes, it establishes intimate contact with that cell. The TC cell (APCs) such as B cells and macrophages, which gather, process,
then releases several pre-formed cytotoxins (molecules lethal to and present exogenous antigens (see figure  15.19b). When a
cells) directly to the target cell. The cytotoxins include perfo- TH cell recognizes a peptide presented by a B cell or macrophage,
rin, a molecule that forms pores in cell membranes, and several it delivers cytokines that activate the cell. Various cytokines are
proteases. Evidence indicates that at the concentrations released also released, depending on the subset of TH cell.
in vivo, perforin simply allows the proteases to enter the target cell.
Once inside, the proteases cause reactions that induce the target The Role of TH Cells in B-Cell Activation
cell to undergo apoptosis. In addition, a specific molecule on the B-cell activation was described earlier (see figure  15.11). This
TC cell can engage a “death receptor” on the target cell, also initiating section reviews that process, but includes more details specific to
apoptosis. Killing the target cell by inducing apoptosis rather than the role of TH cells.

Normal CD8 T-cell receptor

cytoplasmic MHC class I
proteins molecule

All nucleated cells present peptides from TC cells do not recognize peptides
cytoplasmic proteins on MHC class I molecules. presented by healthy “self” cell.

(a)

FIGURE 15.21 Functions

of TC Cells (a) TC cells ignore healthy Virus
“self” cells. (b) TC cells induce apopto- Viral Cytokines
sis in virally infected “self” cells. proteins

? Could a TC cell induce apoptosis in

a “self” cell that lost the ability to
produce MHC class I molecules?
Targeted delivery
of a “death package”

Virally infected “self” cells TC cell recognizes viral peptides Target cell undergoes apoptosis.
present viral peptides on presented by an infected “self” cell
MHC class I molecules. and initiates apoptosis in that target.
It also releases cytokines that alert
neighboring cells.

(b)
 Part III Microorganisms and Humans 373

When a naive B cell binds antigen via its B-cell receptor, normally elicit antibody production. In the body, penicillin can react
the cell takes the antigen in by endocytosis. Proteins within the with proteins, forming a penicillin-protein conjugate. This func-
endosome are then degraded to produce short peptides that can tions in a manner analogous to the H. influenzae conjugate vaccine,
be loaded into the groove of MHC class II molecules. If a TH cell resulting in antibodies that bind penicillin. If IgE antibodies are
encounters a B cell presenting a peptide it recognizes, it delivers formed, their binding to penicillin can result in allergic reactions,
cytokines to that cell. These activate the B cell, allowing it to pro- ruling out the further use of the antimicrobial medication in these
liferate and undergo class switching. The cytokines also drive the individuals. allergy, p. 401 penicillin, pp. 62, 463
formation of memory B cells.
A TH cell does not need to recognize the same epitope as the The Role of TH Cells in Macrophage Activation
B cell it activates. This is because the B cell presents numerous As discussed in chapter 14, macrophages routinely engulf and
peptides from the antigen, and the T-cell receptor could bind any degrade invading microbes, clearing most organisms even before
of those. In fact, a responding B cell probably recognized an epi- an adaptive response is mounted. If this alone is not sufficient to
tope on a pathogen’s surface, whereas a TH cell could very well control the invader, macrophages can be activated by TH cells,
recognize a peptide from within the pathogen. allowing the phagocytes to produce more potent destructive
Understanding the role of antigen processing and presentation mechanisms. macrophages, pp. 340, 348
led to a vaccine that now prevents infection by what was once The steps of macrophage activation are very similar to those
the most common cause of meningitis in children—Haemophilus described for B-cell activation. When macrophages engulf mate-
influenzae. Recall that young children are particularly suscepti- rial, they enclose it within a membrane-bound phagosome
ble to meningitis caused by this organism because it produces a (figure  15.22). The proteins within the phagosome are then
polysaccharide capsule, an example of a T-independent antigen to degraded, and the resulting peptides presented on MHC class II
which this age group responds poorly. A polysaccharide antigen molecules. If a TH cell recognizes one of the peptides, it delivers
can be converted to a T-dependent antigen by covalently attaching cytokines that activate the macrophage. phagosome, p. 72
it to a large protein molecule, making a conjugate vaccine. The When a macrophage is activated, it gets larger, the plasma
polysaccharide component of the vaccine binds to a B-cell recep- membrane becomes ruffled and irregular, and the cell increases its
tor and the entire molecule is taken in. The protein component metabolism so that the lysosomes—which contain antimicrobial
will then be processed and presented to TH cells. Although the B substances—increase in number. The activated macrophage also
cell recognizes the polysaccharide component of the vaccine, the begins producing nitric oxide, a potent antimicrobial chemical,
T cells recognize peptides from the protein component. This rec- along with various compounds that can be released to destroy
ognition leads to B-cell activation and subsequent production of extracellular microorganisms.
antibodies that bind the capsule. Conjugate vaccines against other If the response is still not sufficient to control the infection,
pathogens have also been developed. conjugate vaccines, p. 424 activated macrophages fuse together, forming giant cells. These,
Antigen processing and presentation also explains how along with other macrophages and T cells, can form granulomas
some people develop allergies to penicillin. This medication is a that wall off the offending agent, preventing infectious microbes
hapten, a molecule that binds a B-cell receptor yet does not from escaping to infect other cells. Activated macrophages are an
important aspect of the immune response against diseases such
FIGURE 15.22 The Role as tuberculosis that are caused by organisms capable of surviving
of TH Cells in Macrophage within macrophages. giant cell, p. 348 granuloma, p. 348
Activation
? How does macrophage Subsets of Dendritic Cells and T Cells
activation help prevent
disease? Various subsets of dendritic cells (DC1 and DC2) and effector
helper T cells (TH1, TH2, TH17) steer the immune system toward

1 2 3 CD4
Cytokine delivery T-cell receptor

Secretion
of cytokines

Macrophage Macrophage degrades Peptide fragments TH cell recognizes a presented

engulfs materials. proteins in phagosome are presented on MHC peptide and responds by activating
into peptide fragments. class II molecules. the macrophage. It also releases
cytokines that stimulate TC cells.
374 Chapter 15 The Adaptive Immune Response

an appropriate response. The roles of these subsets are still being their surface; recall that Fc receptors bind the “red flag” portion
clarified, but the different types of dendritic cells produce spe- of antibody molecules. The NK cell attaches to the antibodies and
cific cytokines that cause activated helper T cells to differentiate then delivers perforin- and protease-containing granules directly
into specialized effector subsets. For example, TH1 cells activate to the cell, initiating apoptosis. ADCC, p. 361
macrophages, thereby promoting a response against intracellu- NK cells also recognize and destroy stressed host cells that do
lar pathogens; TH2 cells direct a response against multicellular not have MHC class I molecules on their surface (figure 15.23).
pathogens by recruiting eosinophils and basophils; and TH17 This is important because some viruses have evolved mechanisms
cells recruit neutrophils, thereby directing a response against to dodge the action of cytotoxic T cells by interfering with the
extracellular pathogens. The outcome of some conditions, such as process of antigen presentation; cells infected with such a virus
Hansen’s disease (leprosy), appears to correlate with the type of will essentially be bare of MHC class I molecules and thus can-
helper T-cell response. not be a target of cytotoxic T cells. The NK cells recognize the
lack of MHC class I molecules on those cells, along with certain
MicroAssessment 15.7 molecules that indicate the cells are under stress, and induce the
Dendritic cells expressing co-stimulatory molecules activate T cells
infected cells to undergo apoptosis.
that recognize the presented antigen. TC (CD8) cells recognize antigen Recent evidence indicates that NK cells are more than killing
presented on MHC class I molecules; they induce apoptosis in target machines. For example, they produce cytokines that help regulate
cells and produce cytokines that increase the level of surveillance. TH and direct certain immune responses. Unfortunately, studying
(CD4) cells recognize antigen presented on MHC class II molecules these actions is difficult because there are different subsets of NK
(found on B cells and macrophages); they activate the target cells and cells, and the activities of the various subsets are influenced by
secrete various cytokines that orchestrate the immune response. cues in their local environment.
18. Name three types of antigen-presenting cells.
19. If an effector CD8 cell recognizes antigen presented on an MHC MicroAssessment 15.8
class I molecule, how should it respond?
Natural killer (NK) cells can kill antibody-bound cells by antibody-
20. Why would a person who has AIDS be more susceptible to the dependent cellular cytotoxicity (ADCC). NK cells also kill cells not
bacterium that causes tuberculosis? + bearing MHC class I molecules on their surface.
21. What mechanism do NK cells use to kill “self” cells?
15.8 ■ Natural Killer (NK) Cells 22. Why would a “self” cell not display MHC class I molecules?
23. Why might a virus encode its own version of an MCH class I
Learning Outcome molecule? +
14. Describe two distinct protective roles of NK cells.
15.9 ■ Lymphocyte Development
Natural killer (NK) cells descend from lymphoid progenitor cells,
but lack the antigen-specific receptors that characterize B cells Learning Outcomes
and T cells. Their activities, however, assist the adaptive immune 15. Describe the roles of gene rearrangement, imprecise joining,
responses. lymphoid progenitor cells, p. 338 and combinatorial associations in the generation of diversity.
NK cells induce apoptosis in antibody-bound “self” cells. 16. Describe positive and negative selection of lymphocytes.
This process, antibody-dependent cellular cytotoxicity (ADCC)
allows them to destroy host cells that have viral or other foreign As descendants of hematopoietic stem cells develop into
proteins inserted into their membrane (see figure 15.8). NK cells B cells and T cells, they acquire their ability to recognize distinct
can do this because they have Fc receptors for IgG molecules on epitopes. B cells undergo the developmental stages in the bone

1 MHC class I 2 3 Targeted delivery 4

Virus molecule Viral genome of a “death package”

Natural
killer cell

Virus infects a cell. Virus prevents its host cell Natural killer (NK) cell Infected cell
from displaying MHC class I initiates apoptosis in the undergoes apoptosis.
molecules. Without these, stressed “self” cell that
the cell cannot be a target lacks MHC class I molecules.
of TC cells.

FIGURE 15.23 Natural Killer (NK) Cells Destroy Stressed “Self” Cells That Lack MHC Class I Molecules
? Why would a virus that can interfere with a host cell’s production of MHC class I molecules be at an advantage?
 Part III Microorganisms and Humans 375

marrow; T cells go through the maturation processes described in Generation of Diversity

this section in the thymus. hematopoietic stem cells, p. 338
The events involved in the adaptive immune response, from The mechanisms lymphocytes use to produce a seemingly limit-
the maturation of lymphocytes to the development of their effector less assortment of antibodies and antigen-specific receptors were
functions, are summarized in figure 15.24. first discovered in studies using B cells. Because the processes in

FIGURE 15.24 Summary of the Adaptive Immune Response

? How would the adaptive immune response be affected if Primary lymphoid
memory cells could not be produced? organs

Immature T cells Immature B cells

Peripheral tissues (thymus) (bone marrow)

Dendritic cells that have gathered Secondary lymphoid

antigen in the periphery present it to organs
naive T cells; co-stimulatory molecules
are expressed if the material collected
represents “danger.”

Naive cytotoxic Naive helper Naive B cells

Dendritic cell T cells (CD8) T cells (CD4)
TH cells activate
(gathers antigen
Activation, B cells that present
for presentation Activation,
proliferation, specific antigen
to naive T cells) proliferation,
differentiation to
form effector cells differentiation to
and memory cells form effector cells
and memory cells

TC cells
TH cells

Virus

Memory
TC cells induce apoptosis helper
in infected “self” cells; also T cells
produce cytokines that
alert neighboring cells. Memory
Infected “self” cell cytotoxic Memory
(harbors antigen T cells B cells
within the cell)

TH cells activate macrophages that Plasma cells

present antigen via MHC class II secrete antibodies.
molecules; also produce cytokines that
determine other responses.
Antibodies
(tag extracellular
antigen for removal)

Macrophage (engulfs and destroys Activated

d macrophage
h (engulfs
( and Extracellular
invaders; limited killing powers) destroys invaders; enhanced killing powers) antigen
376 Chapter 15 The Adaptive Immune Response

B cells are very similar to those for T cells, we will use B cells as a mixing and matching different shirts, pants, and shoes, the
general model to describe the generation of diversity with respect traveler can create a wide variety of unique outfits from a lim-
to antigen recognition. ited number of components. Likewise, a B cell expresses three
Each B cell responds to only one epitope, yet the population gene segments, one each from DNA regions called V (variable),
of B cells within the body appears able respond to more than 100 D (diversity), and J (joining), to form an ensemble that encodes
million different epitopes. Based on this number and the informa- a nearly unique variable region of the heavy chain of an antibody
tion presented in chapter 7, it might seem logical to assume that (figure  15.25). Recall that a derivative of this molecule also
the human genome has over 100 million different antibody genes, serves as the B-cell receptor.
each encoding specificity for a single epitope. This is impossible, A human hematopoietic stem cell has about 40 different
however, because the human genome has only 3 billion nucleo- V segments, 25 different D segments, and 6 different J segments
tides and contains about 25,000 genes. in the DNA that encodes the variable region of the heavy chain.
The question of how such tremendous diversity in antibod- As a B cell develops, however, two large regions of DNA are per-
ies could be generated puzzled immunologists until Dr. Susumu manently removed, thereby joining discrete V, D, and J regions.
Tonegawa solved the mystery. For this work, he was awarded a The joined segments encode the heavy chain of the antibody that
Nobel Prize in 1987. the mature B cell is programmed to make. Thus, one B cell could
express the combination V3, D1, and J2 to produce its heavy
Gene Rearrangement chain, whereas another B cell might use V19, D25, and J6; each
A primary mechanism for generating a wide variety of different combination would result in a unique antibody specificity.
antibodies using a limited-size region of DNA employs a Just as DNA segments rearrange in the heavy chain genes
strategy similar to that of a savvy and well-dressed during B-cell development, specific segments move in the light
traveler living out of a small suitcase. By chain genes as well.

Encodes the variable

Encoded by:
region of a heavy chain Encodes the constant
region of a heavy chain
V1 V2 V3 V4 V5 V6 V40 D1 D2 D25 J1 J2 J3 J6
DNA of hematopoietic stem cell
About 40 variable About 25 Six joining region Constant
region gene segments diversity region gene segments region genes
gene segments

b V1 V2 V3 V4 V5 V6 V40 D1 D2 D25 J1 J2 J3 J6
DNA of maturing B cell X;
some DNA is deleted

Segment of DNA Segment of DNA

deleted deleted

Encodes variable region Loss of DNA

of antibody X

V1 V2 V3 D1 J2 J3 J4 J6
DNA of naive B cell X

Transcription and splicing

V3 D1 J2
mRNA of naive B cell X

FIGURE 15.25 Antibody Diversity (a) The variable region of the heavy chain of an antibody molecule is encoded by one each of three
gene segments: V (variable), D (diversity), and J (joining) gene segments. (b) The regions expressed result from loss of DNA as the hematopoietic
stem cell differentiates to become a naive B cell. This diagram shows only the gene segments for the heavy chain and is not drawn to scale.

? Which three aspects of this process in developing B cells contribute to antibody diversity?
 Part III Microorganisms and Humans 377

Imprecise Joining Positive and Negative Selection

As the various segments are joined during gene rearrangement, of Self-Reactive T Cells
nucleotides are often deleted or added between the sections. This
The fate of developing T cells rests on two phases of trials—
imprecise joining changes the reading frame of the encoded pro-
positive and negative selection. Positive selection permits only
tein so that two B cells that have the same V, D, and J segments
those T cells that recognize MHC to develop further. Recall that
for their heavy chain could potentially give rise to antibodies with
the T-cell receptor, unlike the B-cell receptor, recognizes a pep-
very different specificities. Likewise, the segments of the light
tide: MHC complex. The T-cell receptor, therefore, must show
chain often join imprecisely.
at least some recognition of an MHC molecule regardless of the
peptide it is carrying. T cells that show insufficient recognition
Combinatorial Associations
fail positive selection and, as a consequence, are eliminated. Each
Combinatorial association refers to the specific groupings of light T cell that passes positive selection is also subjected to negative
chains and heavy chains that make up the antibody molecule. selection. T cells that recognize “self” peptides presented on MHC
Both types of chains independently acquire diversity through molecules are eliminated. Positive and negative selection proc-
gene rearrangement and imprecise joining. Additional diversity esses are so strict that over 95% of developing T cells undergo
is then introduced when these two molecules join, because the apoptosis in the thymus.
combination of the two chains creates the antigen-binding site (see
figure 15.7b, c).
MicroAssessment 15.9
Mechanisms lymphocytes use to generate diversity of antigen
Negative Selection specificity include rearrangement of gene segments, imprecise joining
of Self-Reactive B Cells of those segments, and combinatorial associations of heavy chains
and light chains. Negative selection eliminates B cells and T cells that
Once a B cell has developed its antigen receptor (B-cell receptor), recognize normal “self” molecules. Positive selection permits only
it passes through rigorous checkpoints in the bone marrow. One of those T cells that recognize the MHC molecules to develop further.
the most important is negative selection, which eliminates any 24. What three gene segments encode the variable region of the
B cell that binds “self.” Most developing B cells fail negative heavy chain of an antibody molecule?
selection and, as a consequence, are induced to undergo apoptosis. 25. Why is negative selection important?
If these cells are not eliminated, then the immune system may 26. How is imprecise joining similar to a frameshift mutation? +
attack “self” substances by mistake.

Summary
15.1 ■ Strategy of the Adaptive Immune Response Primary Lymphoid Organs
As a result of the primary response to an antigen, the secondary Primary lymphoid organs are the sites where B cells and T cells
response is more effective. Humoral immunity works to eliminate mature.
extracellular antigens; cell-mediated immunity (CMI) deals with anti-
gens residing within a host cell. 15.3 ■ The Nature of Antigens
Antigens are molecules that react specifically with an antibody or
Overview of Humoral Immunity lymphocyte; immunogen refers specifically to an antigen that elicits an
Humoral immunity involves B cells (figure 15.1); in response to extra- immune response. The immune response is directed to epitopes (anti-
cellular antigens, B cells proliferate and then differentiate into plasma genic determinants) on the antigen (figure 15.6).
cells that function as antibody-producing factories. Memory B cells are
also formed. 15.4 ■ The Nature of Antibodies

Overview of Cell-Mediated Immunity Structure and Properties of Antibodies (figure 15.7)

Cell-mediated immunity involves T cells; in response to intracellular anti- Antibodies have a Y shape with an antigen-binding site at the end of each
gens, cytotoxic T cells proliferate and then differentiate into TC cells that arm. The tail of the Y is the Fc region. The antibody monomer is com-
induce apoptosis in “self” cells harboring the intruder. Memory cytotoxic posed of two identical heavy chains and two identical light chains. The
T cells are also formed. Helper T cells proliferate and then differentiate to variable region contains the antigen-binding site; the constant region
form TH cells that help orchestrate the various responses of humoral and encompasses the entire Fc region as well as part of the Fab regions.
cell-mediated immunity. Memory helper T cells are also formed.
Protective Outcomes of Antibody-Antigen Binding 
(figure 15.8)
15.2 ■ Anatomy of the Lymphatic System (figure 15.3)
Antibody-antigen binding results in neutralization, opsonization, com-
Lymphatic Vessels plement activation, immobilization and prevention of adherence, cross-
Lymph, which contains antigens that have entered tissues, flows in the linking, and antibody-dependent cellular cytotoxicity (ADCC).
lymphatic vessels to the lymph nodes (figure 15.4).
Immunoglobulin Classes (table 15.1)
Secondary Lymphoid Organs The five major antibody classes—IgM, IgG, IgA, IgD, and IgE—each
Secondary lymphoid organs are the sites at which lymphocytes gather have distinct functions.
to contact antigens.
378 Chapter 15 The Adaptive Immune Response

15.5 ■ Clonal Selection and Expansion of Lymphocytes that detect molecules associated with danger produce co-stimulatory
When an antigen enters a secondary lymphoid organ, only the lym- molecules and are able to activate both subsets of T cells.
phocytes that specifically recognize that antigen will respond; the
Effector Functions of TC (CD8) Cells
antigen receptor they carry on their surface governs this recognition
TC cells induce apoptosis in cells that present peptides they recognize
(figure 15.10). Lymphocytes may be immature, naive, activated, effector,
on MHC class I molecules; they also produce cytokines that allow
or memory cells.
neighboring cells to become more vigilant against intracellular invad-
ers (figure 15.21). All nucleated cells present peptides from endogenous
15.6 ■ B Lymphocytes and the Antibody Response proteins in the groove of MHC class I molecules.
Most antigens are T-dependent antigens, meaning the B cells that
recognize them require help from TH cells. Effector Functions of TH (CD4) Cells (figures 15.11, 15.22)
TH cells activate cells that present peptides they recognize on MHC
B-Cell Activation class II; various cytokines are released, depending on subset of the
B cells present peptides from T-dependent antigens to TH cells for responding TH cell. Macrophages and B cells present peptides from
inspection. If a TH cell recognizes a peptide, it delivers cytokines to the exogenous proteins in the groove of MHC class II molecules.
B cell, initiating the process of clonal expansion, which ultimately gives
rise to plasma cells that produce antibodies (figure 15.11). Subsets of Dendritic Cells and T Cells
Subsets of dendritic cells and TH cells direct the immune system to an
Characteristics of the Primary Response appropriate response.
In the primary response, the expanding B-cell population undergoes
affinity maturation. Under the direction of TH cells, class switching 15.8 ■ Natural Killer (NK) Cells
and memory cell formation also occurs (figures 15.14, 15.15). NK cells mediate antibody-dependent cellular cytotoxicity (ADCC).
Characteristics of the Secondary Response NK cells also induce apoptosis in host cells that are not bearing MHC
Memory cells are responsible for the swift and effective secondary class I molecules on their surface (figure 15.23).
response, eliminating invaders before they cause noticeable harm
(figure 15.12). 15.9 ■ Lymphocyte Development
The Response to T-Independent Antigens Generation of Diversity
T-independent antigens include polysaccharides that have multiple Mechanisms used to generate the diversity of antigen specificity in
identical evenly spaced epitopes, and LPS (figure 15.16). lymphocytes include rearrangement of gene segments, imprecise join-
ing of those segments, and combinatorial associations of heavy and light
15.7 ■ T Lymphocytes: Antigen Recognition chains (figure 15.25).
and Response
Negative Selection of Self-Reactive B Cells
General Characteristics of T Cells (table 15.2, figure 15.19) Negative selection occurs as B cells develop in the bone marrow; cells
Cytotoxic T cells (CD8) recognize antigen presented on major his- to which material binds to their B-cell receptor are induced to undergo
tocompatibility complex (MHC) class I molecules. Helper T cells apoptosis.
(CD4) recognize antigen presented on major histocompatibility com- Positive and Negative Selection of Self-Reactive T Cells
plex (MHC) class II molecules. Positive selection permits only those T cells that show moderate rec-
Activation of T Cells (figure 15.20) ognition of the MHC molecules to develop further. Negative selection
Dendritic cells sample material in tissues and then travel to secondary also occurs.
lymphoid organs to present antigens to naive T cells. The dendritic cells

Review Questions
Short Answer Multiple Choice
1. What is a secondary lymphoid organ? 1. The variable regions of antibodies are located in the
2. Diagram an IgG molecule and label (a) the Fc region and (b) the 1. Fc region. 2. Fab region. 3. light chain.
areas that combine with antigen. 4. heavy chain. 5. light chain and heavy chain.
3. What are the protective outcomes of antibodies binding to antigen? a) 1, 3 b) 1, 5 c) 2, 3 d) 2, 4 e) 2, 5
4. Which antibody class is the first produced during the primary 2. Which of the following statements about antibodies is false?
response? a) If you removed the Fc portion, antibodies would no longer be
5. Which antibody class neutralizes viruses in the intestinal tract? capable of opsonization.
6. Describe clonal selection and expansion in the immune response. b) If you removed the Fc portion, antibodies would no longer be
capable of activating the complement system.
7. How do T-independent antigens differ from T-dependent antigens?
c) If you removed the Fab portion, an antibody would no longer
8. What are antigen-presenting cells (APCs)? be capable of cross-linking antigen.
9. Describe the role of dendritic cells in T-cell activation. d) If IgG were a pentamer, it would bind antigens more
10. How does the role of natural killer cells differ from cytotoxic efficiently.
T cells? e) If IgE had longer half-life, it would protect newborn infants.
 Part III Microorganisms and Humans 379

3. Which class of antibody can cross the placenta? Applications

a) IgA b) IgD c) IgE d) IgG e) IgM 1. Many dairy operations keep cow’s milk for sale and use formula
4. A person who has been vaccinated against a disease should have pri- and feed to raise any calves. One farmer noticed that calves raised
marily which of these types of serum antibodies against that agent on the formula and feed needed to be treated for diarrhea more fre-
2 years later? quently than calves left with their mothers to nurse. He had some
a) IgA b) IgD c) IgE d) IgG e) IgM tests run on the diets and discovered no differences in the calories
5. Which of the following statements about B cells/antibody produc- or nutritional content. The farmer called a veterinarian and asked
tion is false? him to explain the observations. What was the vet’s response?
a) B cells of a given specificity initially have the potential to make 2. What kinds of diseases would be expected to occur as a result of
more than one class of antibody. lack of T or B lymphocytes?
b) In response to antigen, all B cells located close to the antigen
begin dividing. Critical Thinking +
c) Each B cell is programmed to make a single specificity of 1. The development of primary and secondary immune responses
antibody. to an antigen differ significantly. The primary response may take
d) The B-cell receptor allows B cells to detect antigen. a week or more to develop fully and establish memory. The sec-
e) The cell type that makes and secretes antibody is called a ondary response is rapid and relies on the activation of clones of
plasma cell. memory cells. Would it not be better if clones of reactive cells
were maintained regardless of prior exposure? In this way, the
6. Which term describes the loss of specific heavy chain genes?
body could always respond rapidly to any antigen exposure. Would
a) Affinity maturation there be any disadvantages to this approach? Why?
b) Apoptosis 2. Early investigators proposed two hypotheses to explain the speci-
c) Clonal selection ficity of antibodies. The clonal selection hypothesis states that each
d) Class switching lymphocyte can produce only one specificity of antibody. When
7. Which of the following specifically refers to an effector lympho- an antigen binds to that B-cell receptor, the lymphocyte is selected
cyte? to give rise to a clone of plasma cells producing the antibody. The
a) B cell b) Cytotoxic T cell template hypothesis states that any antigen can interact with any
c) Helper T cell d) Plasma cell lymphocyte and act as a template, causing newly forming antibod-
ies to be specific for that antigen. In one experiment to test these
8. Which markers are found on all nucleated cells?
hypotheses, an animal was immunized with two different antigens.
a) MHC class I molecules After several days, lymphocytes were removed from the animal
b) MHC class II molecules and individual cells placed in separate small containers. Then, the
c) CD4 original two antigens were placed in the containers with each cell.
d) CD8 What result would support the clonal selection hypothesis? The
9. Which of the following are examples of an antigen-presenting cell template hypothesis?
(APC)?
1. Macrophage 2. Neutrophil 3. B cell
4. T cell 5. Plasma cell
a) 1, 2 b) 1, 3 c) 2, 4 d) 3, 5 e) 1, 2, 3
10. What is the appropriate response when antigen is presented on
MHC class II molecules?
a) An effector CD8 cell should kill the presenting cell.
b) An effector CD4 cell should kill the presenting cell.
c) An effector CD8 cell should activate the presenting cell.
d) An effector CD4 cell should activate the presenting cell.
16 Host-Microbe Interactions
KEY TERMS
Acute Infection An infection
characterized by symptoms that
develop fairly quickly and last a
relatively short time.
Infectious Disease An infection
that results in noticeable impairment
of body function.
Latent Infection Infection in
Chronic Infection An infection which the infectious agent is present
that generally develops slowly and but not causing symptoms.
lasts for months or years.
Normal Microbiota The
Colonization Establishment and population of microorganisms
growth of a microorganism on a routinely found growing on the body
surface. surfaces of healthy individuals.
Endotoxin The lipopolysaccharide Opportunistic Pathogen
(LPS) component of the outer A microbe that causes disease
membrane of Gram-negative only when introduced into an
bacteria; lipid A is responsible for unusual location or into an
the toxic properties of LPS. immunocompromised host.
Exotoxin A toxic protein produced Primary Pathogen A microbe
by a microorganism; often simply able to cause disease in an otherwise
referred to as a toxin. healthy individual.
Immunocompromised A host Virulence Factors Traits of a
with a weakness or defect in the microbe that promote pathogenicity.
innate or adaptive defenses.
Infection Colonization by a
Salmonella enterica serotype Typhimurium invading cultured human cells (color- pathogen on or within the body.
enhanced scanning electron micrograph).

E 
very day we contact an enormous number and variety of
A Glimpse of History microorganisms. Some enter our respiratory system as we
The ancients thought epidemics and diseases were divine punishment of breathe; other are ingested with each bite of food or sip of
the people for their sins. By the time of Moses, however, the Egyptians drink; and still more adhere to our skin whenever we touch an
and Hebrews had come to believe that leprosy could be transmitted by object or surface. It is important to recognize, however, that the
contact with lepers. In Europe, around 430 b.c. Thucydides had con-
vast majority of these microbes generate no ill effects whatsoever.
cluded that some plagues were contagious. By the Middle Ages, many
Some may colonize the body surfaces, taking up residence with
accepted this, and fled cities to escape the diseases. Fracastorius, in 1546,
first proposed that communicable diseases were caused by living agents the variety of other harmless microbes that live there; others are
passed from one person or animal to another. He had no way to test this sloughed off with dead epithelial cells. Most of those swallowed
idea, however. are either killed in the stomach or eliminated in feces.
With Leeuwenhoek’s discovery of microorganisms in the late sev- Relatively few microbes cause noticeable damage to the human
enteenth century, people began to suspect that microorganisms might body, such as invading tissues or producing toxic substances. Those
cause disease, but the techniques of the times could not prove this. It that can are called pathogens. They have distinct characteristics
was not until 1876 that Robert Koch offered convincing proof of what that allow them to avoid at least some of the body’s defenses.
is now known as the germ theory of disease. He showed that Bacillus Research into how these microbes evade our innate and adaptive
anthracis causes anthrax, an often fatal disease of humans, sheep, and defenses is unraveling an impressive array of ploys. The knowledge
other animals. With his microscope, he observed B. anthracis cells in the
we are gaining in areas such as genomics and immunology have
blood and spleen of dead sheep. He then inoculated mice with the infected
given new insights into the pathogenic strategies, fueling hope that
sheep blood, and recovered B. anthracis from the blood of those mice. In
addition, he grew the bacteria in pure culture and showed that they caused therapies targeted to specific pathogens can be developed.
anthrax when injected into healthy mice. From these experiments and This chapter will explore some of the ways in which microbes
later work with Mycobacterium tuberculosis, Koch formalized a group colonize the human host, living either as members of the normal
of criteria for establishing the cause of an infectious disease, known as microbiota or causing disease. It will also describe how pathogens
Koch’s postulates. evade or overcome the immune responses and damage the host.

380
 Part III Microorganisms and Humans 381

MICROBES, HEALTH, AND DISEASE

Many people think of microorganisms as “germs” that should rou- 16.2 ■ The Normal Microbiota
tinely be killed or avoided. Most microbes are harmless, however,
and many are beneficial. The organisms that routinely reside on Learning Outcomes
the body’s surfaces are the normal microbiota, or normal flora.
2. Describe three protective roles of the normal microbiota.
This relationship is a delicate balancing act, though, because
3. Describe how the composition of the normal microbiota can
some members of the normal microbiota, as well as microbes that
change over time.
make incidental contact with humans, can cause disease if the
opportunity arises. Weaknesses or defects in the innate or adaptive
The normal microbiota is the population of microorganisms
defenses can leave people vulnerable to invasion; these individu-
routinely found growing on the body of healthy individuals
als are said to be immunocompromised. Factors that can lead to
(figure  16.1). Microbes that typically inhabit body sites for
an individual becoming immunocompromised include malnutri-
extended periods are resident microbiota, whereas temporary
tion, cancer, AIDS or other diseases, surgery, wounds, genetic
occupants are transient microbiota. Considering how important
defects, alcohol or drug abuse, and immunosuppressive therapy
this population is to human health, relatively little is known about
that accompanies procedures such as organ transplants.
its members. With that in mind, the Human Microbiome Project
is aimed at studying this diverse population. Human Microbiome
16.1 ■ The Anatomical Barriers Project, p. 225
as Ecosystems
MicroByte
Learning Outcome There are more bacteria in just one person’s mouth than there are
people in the world!
1. Compare and contrast mutualism, commensalism, and parasitism.

The skin and mucous membranes are barriers against invading

microorganisms, but they also host a complex ecosystem—an
interacting biological community. The intimate relationships
between the microorganisms and the human body are an example
of symbiosis, meaning “living together.”
Microorganisms can have a variety of symbiotic relationships
with each other and with the human host. These relationships may
take on different characteristics depending on the closeness of the Nose
Staphylococcus Mouth
association and the relative advantages to each partner. Symbiotic Corynebacterium Streptococcus
associations can be one of several forms, and these may change, Fusobacterium
Throat Actinomyces
depending on the state of the host and the traits of the microbes: Streptococcus Leptotrichia
Moraxella Veillonella
■ Mutualism is an association in which both partners benefit. Corynebacterium
In the large intestine, for example, some bacteria synthesize Haemophilus
Skin
Neisseria
vitamin K and certain B vitamins. These nutrients are then Staphylococcus
Mycoplasma
available for the host to absorb. The bacteria residing in the Propionibacterium
intestine benefit as well, supplied with warmth and a variety Large intestine
of different energy sources. Bacteroides
Escherichia
■ Commensalism is an association in which one partner ben- Proteus
efits but the other remains unharmed. Many microbes living Klebsiella
Lactobacillus
on the skin are neither harmful nor helpful to the human host, Streptococcus
but they obtain food and other necessities from the host. Candida
Clostridium
■ Parasitism is an association in which one organism, the parasite, Pseudomonas
benefits at the expense of the other. All pathogens are parasites, Enterococcus
but medical microbiologists often reserve the word parasite for Urethra Vagina
eukaryotic pathogens such as protozoa and helminths. Streptococcus Lactobacillus
Mycobacterium
MicroAssessment 16.1 Escherichia
Bacteroides
Depending on the relative benefit to each partner such as a human
host and a microbe, the relationship can be described as mutualism, FIGURE 16.1 Normal Microbiota This shows only some of
commensalism, or parasitism. the common genera; many others may also be present.

1. How is mutualism different from commensalism? ? What can happen if members of the normal microbiota in the
intestinal tract are killed or their growth suppressed?
382 Chapter 16 Host-Microbe Interactions

The Protective Role of the Normal of the Firmicutes, a phylum that includes Clostridium and Bacillus
Microbiota species, whereas thin individuals typically have more members of
the Bacteroidetes, a phylum that includes Bacteroides species. As
One of the most significant contributions of the normal microbiota obese people lost weight, their intestinal microbiota changed to
to health is protection against pathogens. As discussed in chapter resemble that of typically lean people. A variety of studies to track
14, the normal microbiota excludes pathogens by (1) covering changes in the normal microbiota in both health and disease are
binding sites that might otherwise be used for attachment, (2) con- now underway as part of the Human Microbiome Project.
suming available nutrients, and (3) producing compounds toxic to
other bacteria. When members of the normal microbiota are killed MicroAssessment 16.2
or their growth suppressed, as can happen during antibiotic treat-
ment, pathogens may colonize and cause disease. For instance, The normal microbiota provides protection against potentially
harmful organisms and stimulates the immune system.
certain antibiotics inhibit the Lactobacillus species that normally
predominate in the vagina of mature females. These bacteria 2. What factor favors abundant growth of Clostridium difficile in
normally suppress the growth of the yeast Candida albicans, and the intestine?
without their protective action, the yeast cells can multiply to high 3. Why would the immune response to members of the normal
numbers, resulting in vulvovaginal candidiasis. Oral antibiotics can microbiota cross-react with pathogens? +
also inhibit members of the normal intestinal microbiota, allowing
the overgrowth of toxin-producing strains of Clostridium difficile
that cause antibiotic-associated diarrhea and colitis. vulvovaginal 16.3 ■ Principles of Infectious
candidiasis, p. 618 Clostridium difficile–associated disease, p. 594 Disease
Another critical role of the normal microbiota is to stimulate the
adaptive immune system. The importance of this can be shown in Learning Outcomes
mice reared in a microbe-free environment. These “germ-free” ani- 4. Define the terms primary pathogen, opportunist, and virulence.
mals have greatly underdeveloped mucosa-associated lymphoid tissue 5. Describe the characteristics of infectious diseases, including the course
(MALT). In addition, antibodies produced against members of the of disease, duration of symptoms,and distribution of the pathogen.
normal microbiota bind to pathogen surfaces as well. MALT, p. 358
The normal microbiota appears to play an important role in The term colonization refers to a microbe establishing itself and
the development of oral tolerance by the immune system. In a multiplying on a body surface. If the microbe has a parasitic rela-
complex series of events, our defenses learn to lessen the immune tionship with the host, then the term infection can be used. That
response to the many microbes that routinely inhabit the gut, as is, a member of the normal microbiota is said to have colonized
well as foods that pass through. Recent studies into the actions the host, but a pathogen is described as having either colonized or
of regulatory T cells indicate that early and consistent exposure infected the host. Infection does not always lead to illness. It can
to certain microbes in the gut stimulates these T cells, thereby be subclinical, meaning that symptoms either do not appear or are
preventing the immune system from overreacting to harmless mild enough to go unnoticed.
microbes and substances. This idea is the basis of the hygiene An infection that results in disease (a noticeable impairment
hypothesis, which proposes that insufficient exposure to microbes of body function) is called an infectious disease. Diseases are
can lead to allergies. It is a fine balance, however, because contact characterized by symptoms and signs; symptoms are the subjec-
with certain pathogens can be deadly. tolerance, p. 355 tive effects of the disease experienced by the patient, such as pain
and nausea, whereas signs are the objective evidence, such as
The Dynamic Nature of the Normal rash, pus formation, and swelling.
Microbiota Effects of one disease may leave a person predisposed to
developing another. For example, a respiratory illness that dam-
A healthy human fetus is sterile until the protective membrane that ages the mucociliary escalator makes a person more likely to
surrounds it ruptures just before birth. During the passage through develop pneumonia. The initial infection is a primary infection;
the birth canal, the baby is exposed to a variety of microbes that an additional infection that occurs as a result of the primary infec-
take up residence on its skin and in the digestive tract. Various tion is a secondary infection. mucociliary escalator, p. 337
microorganisms in food, on other humans, and in the environment
soon also become established as residents on the newborn.
The composition of the normal microbiota is dynamic. At
Pathogenicity
any one time, the makeup of this complex ecosystem represents a A primary pathogen, or more simply, a pathogen, is a microbe
balance of many forces that can alter the microbial population’s or virus that causes disease in otherwise healthy individuals.
quantity and composition. Changes occur in response to physi- Diseases such as plague, malaria, measles, influenza, diphtheria,
ological variations within the host (such as hormonal changes), tetanus, and tuberculosis are caused by primary pathogens.
and as a direct result of the activities of the human host (such as An opportunistic pathogen, or opportunist, causes disease
consuming food). An intriguing example of the dynamic nature of only when the body’s innate or adaptive defenses are compro-
the microbiota was the discovery that the intestinal microbiota of mised, or when introduced into an unusual location. Opportunists
obese and lean people differs. Obese people have more members can be members of the normal microbiota or they can be
 Part III Microorganisms and Humans 383

common in the environment. For instance, Pseudomonas species vague symptoms such as malaise and headache. After the illness
are environmental bacteria that routinely come into contact with subsides, there is a period of convalescence, the stage of recu-
healthy individuals without harmful effect, yet they can cause peration and recovery from the disease. Even though there is no
fatal infections in individuals who have the genetic disease cystic indication of infection during the incubation and convalescent
fibrosis and also in burn patients (see figure 23.6). Ironically, as periods, many infectious agents can still be spread during these
our healthcare systems improve, extending the life span of patients stages. Some individuals, called carriers, harbor an infectious
through surgery and immunosuppressive drugs, diseases caused agent for months or years and continue to spread the pathogen,
by opportunists are becoming more common. Also, many organ- even though they show no signs or symptoms of the disease.
isms not previously known to cause disease have now been shown The impact of carriers on spread of disease will be discussed in
to do so in severely immunocompromised patients. chapter 19. carriers, p. 439
The term virulence refers to the degree of pathogenicity of Following recovery from infection, or after immunization, the
an organism. An organism described as highly virulent is more host normally has accumulated protective antibodies and memory
likely to cause disease, particularly severe disease, than might oth- lymphocytes that prevent reinfection with the same microbe. In
erwise be expected. Streptococcus pyogenes causes strep throat, most cases, the host is no longer susceptible to infection with that
for example, but certain strains are particularly virulent, causing particular infectious agent.
diseases such as necrotizing fasciitis (“flesh-eating disease”).
Virulence factors are the traits of a microorganism that specifi- Duration of Symptoms
cally allow it to cause disease. The genes encoding these traits can Infections and the associated diseases are often described accord-
sometimes be transferred horizontally. necrotizing fasciitis, p.  553 ing to the timing and duration of the symptoms (figure 16.2):
horizontal gene transfer, p. 200
■ Acute infections are characterized by symptoms that develop
quickly but last only a short time; an example is strep throat.
Characteristics of Infectious Disease
■ Chronic infections develop slowly and last for months or
Infectious diseases that spread from one host to another are years; an example is tuberculosis.
called communicable, or contagious, diseases. Some contagious
■ Latent infections are never completely eliminated; the
diseases, such as colds and measles, are easily transmitted. The
microbe continues to exist in host tissues, often within host
ease with which a contagious disease spreads partly reflects the
cells, without causing any symptoms. If there is a decrease in
infectious dose—the number of microbes necessary to establish
immunity, the latent infection may become reactivated and
an infection. For example, the intestinal disease shigellosis is quite
symptomatic. Note that the symptomatic phase of the disease
contagious in humans because only 10 to 100 cells of a Shigella
may be either acute or chronic. For example, the infection
species need be ingested to establish an infection; in contrast,
caused by the varicella-zoster virus results in the characteris-
salmonellosis does not spread as readily because as many as 106
tic symptoms of chickenpox, an acute illness. The illness is
cells of Salmonella enterica serotype Enteritidis must be ingested
stopped by an effective immune response, leaving the host
to cause illness. The difference in these infectious doses reflects,
immune to reinfection. The virus, however, is not completely
in part, the pathogen’s ability to survive the acidic conditions
eliminated. It takes refuge in sensory nerves, held in check by
encountered as the cells pass through the stomach. Generally, the
the immune system. Later in life, infectious viral particles
infectious dose is expressed as the ID50, an experimentally derived
may be produced again, causing the skin disease shingles
figure  that indicates the number of microbial cells administered
(herpes zoster). In tuberculosis, the mycobacteria are often
that resulted in disease in 50% of the test population. Shigella,
p. 588 Salmonella enterica serotype Enteritidis, p. 592

Course of Infectious Disease Incubation period Illness Convalescence

The course of an infectious disease includes several
stages (figure  16.2). The time between introduction of Acute. Illness is short term because the pathogen is eliminated by the host
defenses; person is usually immune to reinfection.
a microbe to a susceptible host and the onset of illness
is the incubation period. This varies considerably,
from only a few days for the common cold, to several Incubation period Illness (long lasting)
weeks for hepatitis A, to many months for rabies, and
Chronic. Illness persists over a long time period.
even years for Hansen’s disease (leprosy). The length
of the incubation period depends on a variety of factors,
including the growth rate of the pathogen, the host’s Incubation period Illness Convalescence Latency Recurrence
condition, and the number of infectious cells or virions
Latent. Illness may recur if immunity weakens.
encountered. Hansen’s disease, p. 650
A phase of illness follows the incubation period. FIGURE 16.2 The Course of Infectious Diseases Infections can be acute,
During this period, a person will experience the signs chronic, or latent.
and symptoms of the disease. In some cases, onset of ? Some diseases include a prodromal phase. Where would this phase fit in the
illness is heralded by a prodromal phase—the early, figure?
384 Chapter 16 Host-Microbe Interactions

initially confined within a small area by host defenses, caus-

ing no symptoms; much later, the bacteria may begin multi-
plying again, resulting in a chronic illness. Other diseases in
which the causative agent becomes latent include cold sores
and genital herpes. chickenpox, p. 534 tuberculosis, p. 502

Distribution of the Pathogen

Infections are often described according to the distribution of
the causative agent in the body. In a localized infection, the 1 The microorganism must be present in every case of the disease, but
microbe is limited to a small area; an example is a boil caused not in healthy hosts.
by Staphylococcus aureus. In a systemic infection, the infectious
agent is disseminated (spread) throughout the body; an example
is measles.
The suffix -emia means “in the blood.” Thus, bacteremia indi-
cates that bacteria are circulating in the bloodstream. Note that this
term does not necessarily imply a disease state. A person can become
transiently bacteremic after vigorous tooth brushing. Toxemia
indicates that toxins are circulating in the bloodstream. The organism 2 The microorganism must be grown in pure culture from diseased hosts.
that causes tetanus, for instance, produces a localized infection yet its
toxins circulate in the bloodstream. The term viremia indicates that
viral particles are circulating in the bloodstream.

MicroAssessment 16.3
A primary pathogen can cause disease in an otherwise healthy
individual; an opportunist causes disease in an immunocompromised
host. The course of infectious disease includes an incubation period,
illness, and a period of convalescence. Infections can be acute or
chronic, latent, localized, or systemic.
4. Why are diseases caused by opportunists becoming more 3 The same disease must be produced when a pure culture of the
frequent? microorganism is introduced into susceptible hosts.

5. Give an example of a microbe that causes a latent infection.

6. What factors might contribute to a long incubation period? +

16.4 ■ Establishing the Cause

of Infectious Disease
Learning Outcome 4 The same microorganism must be recovered from the experimentally
infected hosts.
6. List Koch’s postulates, and compare them to the Molecular Koch’s
postulates.
FIGURE 16.3 Koch’s Postulates These criteria provide a founda-
tion for establishing that a given microbe causes a specific disease.
Criteria are needed to guide scientists as they try to determine the
cause of infectious diseases. They can also be helpful when study- ? Why cannot Koch’s postulates be used to show that Treponema
pallidum causes syphilis?
ing the disease process.

Koch’s Postulates 3 The same disease must be produced when a pure culture of
Koch’s postulates—the criteria that Robert Koch used to estab- the organism is introduced into susceptible hosts.
lish that Bacillus anthracis causes anthrax (see A Glimpse of 4 The organism must be recovered from the experimentally
History)—provide a foundation for establishing that a given infected hosts.
microbe causes a specific infectious disease (figure 16.3):
When Koch studied anthrax, he grew B. anthracis from all cases
1 The microorganism must be present in every case of the disease. examined; he introduced pure cultures of the organisms into
2 The organism must be grown in pure culture from diseased hosts. healthy susceptible mice, again causing the disease anthrax.
 Part III Microorganisms and Humans 385

Finally, he recovered the organism from the experimentally study of pathogens such as E. coli and Streptococcus pyogenes,
infected mice. which can cause several different diseases depending on the viru-
It is important to note that there are many situations in which lence factors of a given strain. Molecular Koch’s postulates are
Koch’s postulates cannot be carried out. For example, the second as follows:
postulate cannot be fulfilled for organisms that cannot be grown
1. The virulence factor gene or its product should be found in
in laboratory medium, such as Treponema pallidum (causes
pathogenic strains of the organism.
syphilis). In other cases, the third postulate does not always hold
true. There are many examples, including cholera and polio, in 2. Mutating the virulence gene to disrupt its function should
which some infected people do not have symptoms of disease. In reduce the virulence of the pathogen.
addition, some diseases are polymicrobial, meaning that multiple 3. Reversion of the mutated virulence gene or replacement with
species act together to cause the illness; an example is chronic a wild-type version should restore virulence to the strain.
periodontal disease. Also, suitable experimental animal hosts
As with the traditional Koch’s postulates, it is not always pos-
are not available for some diseases and it would not be ethical
sible to apply all of these criteria, but they provide an approach to
to test the postulates on humans because of safety concerns.
studying how infectious agents cause disease.
Nevertheless, despite the limitations of the postulates, they have
provided scientists with a logical framework for determining the
causes of infectious diseases. syphilis, p.  626 cholera, p.  586 MicroAssessment 16.4
polio, p. 656 periodontal disease, p. 577
Koch’s postulates can be used to establish that a given microbe causes
a specific infectious disease. Molecular Koch’s postulates are used to
identify the virulence factors responsible for disease.
Molecular Koch’s Postulates
7. How were Koch’s postulates used to prove the cause of anthrax?
Molecular Koch’s postulates are similar in principle to Koch’s
8. Why can Koch’s postulates not be used to identify the causes of
postulates, but they rely on molecular techniques to study a diseases due to polymicrobial infections? +
microbe’s virulence factors. They are particularly relevant in the

MECHANISMS OF PATHOGENESIS
From a microbe’s perspective, the interior of the human body ■ Invasion of host tissues. The microbe penetrates the first-line
is a rich source of nutrients guarded by the innate and adap- defenses and then multiplies within the tissues. Organisms
tive defenses. The ability to get past these defenses and cause that do this generally have mechanisms to avoid destruction
damage is what distinguishes pathogens from other microbes. by macrophages; some also have mechanisms to avoid anti-
Understanding how they do this helps illustrate why only certain bodies. There are numerous examples of bacteria that invade,
microbes can cause disease in a healthy host. Pathogenic mecha- including Mycobacterium tuberculosis (causes tuberculosis),
nisms generally follow one of several patterns: Yersinia pestis (causes plague) and Salmonella enterica
(most strains cause gastroenteritis and one causes typhoid
■ Production of toxins that are then ingested. The microbe
fever). Mycobacterium tuberculosis, p. 503 Yersinia pestis, p. 678
does not grow on or in the host, so this is not an infection
but rather a foodborne intoxication, a form of food poison- ■ Invasion of host tissues, followed by toxin production.
ing. The only virulence determinant is toxin production. These microbes are similar to those in the previous cat-
Relatively few bacteria cause foodborne intoxication; these egory, but in addition to invading, they also make toxins.
include Clostridium botulinum (causes botulism), and toxin- Examples include Shigella dysenteriae (causes diarrhea) and
producing strains of Staphylococcus aureus (cause staphy- Clostridium tetani (causes tetanus). Clostridium tetani, p. 555

lococcal food poisoning). botulism, p.  652 Staphylococcus A successful pathogen needs only to overcome the host
aureus foodborne intoxication, p. 757 defenses long enough to multiply and then exit the host. In fact,
■ Colonization of mucous membranes of the host, followed a pathogen that completely overwhelms the host defenses is actu-
by toxin production. The microbe adheres to a mucous ally at a disadvantage because it will likely kill the host. If the
membrane such as the lining of the intestinal or upper respira- host dies, the pathogen loses an exclusive source of nutrients and
tory tracts and multiplies to high numbers. There, it produces perhaps the opportunity to be transmitted.
a toxin that interferes with cell function. Examples of bacteria Pathogens and their hosts generally evolve over time to a
that do this include Vibrio cholerae (causes cholera), E. coli state of balanced pathogenicity. The pathogen becomes less
O157:H7 (causes bloody diarrhea), and Corynebacterium virulent while the host becomes less susceptible. This was demon-
diphtheriae (causes diphtheria). cholera, p.  586 E. coli strated when the myxoma virus was intentionally introduced into
O157:H7 diarrhea, p. 590 diphtheria, p. 490 Australia in the early 1950s to kill the rapidly increasing rabbit
386 Chapter 16 Host-Microbe Interactions

population. As expected, the rabbit popula-

tion dropped dramatically after the virus was
introduced. Eventually, however, the numbers
of rabbits again began rising. Viruses isolated Pili with
from these rabbits were shown to be less viru- adhesins Bacterial cell
lent than the original strain, and the rabbits were
more resistant to the original virus strain.
The next sections will describe how pathogens
adhere to and colonize host tissue, avoid innate defenses,
avoid adaptive defenses, and cause the damage associated
with disease. We will focus on mechanisms of bacterial pathogene-
sis because these are by far the most thoroughly characterized; later
in the chapter, we will discuss pathogenesis of viruses and eukary-
otic organisms. As we describe various virulence factors, recognize
that their roles are not mutually exclusive—a single structure can Receptor
serve more than one purpose. Also note that one microbe can have
more than one virulence factor, and various strains of the same spe-
cies can have different virulence factors.

Host cell
16.5 ■ Establishing Infection
FIGURE 16.4 Pili Attachment to Host Cell An adhesin at the tip
Learning Outcomes of a pilus attaches to a molecule on the host cell surface.
7. Describe the requirements for adherence and colonization.
? What would happen if a pathogen lost the ability to produce
8. Explain the role of type III secretion systems in infection. adhesins?

To cause disease, most pathogens must first adhere to a body sur-

face and then multiply. In some cases, they deliver molecules to Strains that cause urinary tract infections have pili that attach to
epithelial cells, causing changes in those cells. the bladder, and strains that cause watery diarrhea have pili that
adhere to cells of the small intestine.

Adherence
The first-line defenses are very effective in sweeping microbes
Colonization
away, so pathogens must adhere to host cells to initiate infection. A microorganism must multiply in order to colonize the host. In
Microbes that attach to cells, however, do not necessarily cause many cases, pathogens grow in biofilms. biofilms, p. 84
disease. For example, members of the normal microbiota often To colonize a mucosal surface, the pathogen must deal with
adhere to epithelial cells with no ill effect whatsoever. Other fac- the host’s defenses that protect those surfaces. Recall, for exam-
tors such as toxin production or invasion generally must come into ple, that the body uses lactoferrin and transferrin to bind iron,
play before disease results. first-line defenses, p. 336 thereby limiting the growth of microbes. Some pathogens respond
Bacteria use adhesins to attach to host cells (figure  16.4). by producing their own iron-binding molecules, siderophores;
These are often located at the tips of pili (pili used for attachment others can use the iron bound to the host proteins. lactoferrin and
are often called fimbriae; see figure 3.40). Adhesins can also be a transferrin, p. 337
component of other surface structures such as capsules or various Secretory IgA also protects mucosal surfaces. Pathogens,
cell wall proteins (see figure 3.35). pili, p. 65 capsule, p. 62 however, have evolved mechanisms to avoid those antibodies.
The molecule to which an adhesin attaches is called the Mechanisms include rapid turnover of pili (to shed any bound
receptor. Note that receptors have distinct roles for the host cells; antibody), antigenic variation, and IgA proteases (enzymes that
the microbes merely exploit the molecules for their own use. cleave IgA antibodies). IgA, p. 362 antigenic variation, p. 178
For example, the normal role of the receptor used by Neisseria If the body site has normal microbiota, the new arrival must
gonorrhoeae is to help protect host cells from damage by the compete for space and nutrients. It must also tolerate any toxic
complement system. Receptors are typically glycoproteins or gly- products such as fatty acids produced by the competitors.
colipids, and the adhesin binds to the sugar portion. complement
system, p. 344 glycoprotein, p. 29
Adhesin-receptor binding is highly specific, dictating the
Delivering Effector Proteins
type of cells to which the bacterium can attach. For instance, the to Host Cells
adhesin of common E. coli strains allows them to adhere to cells Some Gram-negative pathogens deliver proteins directly into host
that line the large intestine, where the strains multiply as part of cells using secretion systems. For example, a type III secretion
the normal microbiota. Pathogenic E. coli strains have additional system, or injectisome, is a syringelike structure that injects pro-
adhesins, broadening the range of tissues to which they can attach. teins into eukaryotic cells (figure  16.5). The injected proteins,
 Part III Microorganisms and Humans 387

Effector
Penetrating the Skin
Bacterial Skin is the most difficult anatomical barrier for microbes to
cytoplasm penetrate. Bacterial pathogens that invade via this route rely on
skin-damaging injury. Staphylococcus aureus enters tissues via a
cut or other wound. Yersinia pestis is injected by infected fleas.
plague, p. 678

Bacterial
periplasm Penetrating Mucous Membranes
Mucous membranes are the entry points for most pathogens,
but the invasive processes are complex and difficult to study. It
appears, however, that there are at least two mechanisms used for
invasion: directed uptake by cells and exploiting antigen-sampling
processes.

Host cell Directed Uptake by Cells

Some pathogens induce non-phagocytic cells to engulf them.
The pathogen first attaches to a cell, then triggers the process
of endocytosis. endocytosis, p. 72
FIGURE 16.5 Type III Secretion Systems Gram-negative bacte- Gram-negative bacteria often inject effector proteins
ria use type III secretion systems to deliver certain molecules directly
to host cells, inducing changes in those cells. Peptidoglycan is not that induce engulfment by host cells. Salmonella species,
shown in this figure. for example, use a type III secretion system to deliver spe-
cific proteins to intestinal epithelial cells. These cause actin
? What bacterial structure do type III secretion systems resemble?
molecules in the host cell cytoplasm to rearrange, resulting
in characteristic membrane ruffling on the cell’s surface
(figure 16.6). The ruffles enclose the bacterial cells, bringing
referred to as effector proteins, induce changes such as altering them into the intestinal cell.
the cell’s cytoskeleton structure. Some effector proteins direct the
host cell to engulf the bacterial cell, a process discussed in the next
section. Several types of secretion systems have been discovered,
and some can inject molecules other than proteins. secretion, p. 56
cytoskeleton, p. 73 Ruffle M-cell surface

MicroAssessment 16.5
Pathogens use adhesins, often on pili, to bind to a body surface. To
colonize a surface, the pathogen must often compete with the normal
microbiota, prevent binding of secretory IgA, and obtain iron. Some
bacteria deliver effector proteins to epithelial cells, inducing a specific
change in those cells.
9. What are siderophores?
10. What is a type III secretion system?
11. Why is it a good strategy for a microbe to adhere to a receptor
that plays a critical function for a host cell? +

16.6 ■ Invasion—Breaching
the Anatomical Barriers
Learning Outcome
9. Describe the mechanisms pathogens use to penetrate the skin and
mucous membranes.
Bacterial cell 10 μm

Some bacterial pathogens cause disease while remaining on the FIGURE 16.6 Ruffling Salmonella enterica serotype Typhimurium
mucosal surfaces, but many others penetrate the anatomical bar- inducing ruffles on an M cell (a specialized epithelial cell), leading to
riers. By crossing the epithelial barrier, invading microbes can uptake of the bacterial cells.
multiply in the nutrient-rich tissues without competition. ? How do Salmonella cells induce ruffling?
388 Chapter 16 Host-Microbe Interactions

Exploiting Antigen-Sampling Processes MicroAssessment 16.6

Recall that mucosa-associated lymphoid tissue (MALT) samples Skin is the most difficult barrier for microbes to penetrate. Some
material from the mucosal surface. Some pathogens use this pro- pathogens induce mucosal epithelial cells to engulf the bacterial cells.
cess to cross the membranes. MALT, p. 358 Some take advantage of antigen-sampling processes.
Several pathogens use M cells to cross the intestinal barrier. 12. How do Shigella species enter intestinal epithelial cells?
Recall that M cells transport material from the lumen of the intes-
13. Why does Mycobacterium tuberculosis direct macrophages to
tine to the Peyer’s patches (see figure 15.5). Most microbes deliv- engulf them?
ered this way are destroyed by the macrophages in the Peyer’s
14. Why would it be difficult to study invasion of mucous
patches, but pathogens have mechanisms to avoid this fate. When membranes? +
Shigella cells are transferred to the macrophages, for instance, the
bacteria survive, and eventually induce the phagocyte to undergo
apoptosis (figure  16.7). The freed bacterial cells then bind to
the base of the mucosal epithelial cells and cause these non-
16.7 ■ Avoiding the
phagocytic cells to engulf them, using a mechanism similar to that Host Defenses
of Salmonella. M cell, p. 358 Peyer’s patches, p. 358
Some pathogens invade by means of alveolar macrophages, Learning Outcome
which engulf material that enters the lungs. Mycobacterium 10. Describe mechanisms that bacteria use to avoid complement
tuberculosis produces surface proteins that direct their uptake by system proteins, antibodies, and destruction by phagocytes.
macrophages. Although this might seem to be a disadvantage to
the bacteria, it actually allows them to avoid a process that could Inside the body, invading microorganisms soon encounter the
otherwise lead to macrophage activation. Mycobacterium cells innate and adaptive immune defenses. Pathogens as a group have
survive within macrophages that have not been activated. evolved a variety of mechanisms to avoid the otherwise lethal
effects of these defenses.
3 Within an epithelial cell, Shigella cells cause
the host actin to polymerize. This propels Hiding Within a Host Cell
the bacterial cell, sometimes with enough
force to push it into the next cell. Some pathogens enter host cells, where they hide from comple-
ment proteins, phagocytes, and antibodies. Once a Shigella cell
Lumen of the intestine Mucous is within an intestinal epithelial cell, it directs its own transfer to
membrane adjacent cells (figure 16.7). It does this by causing the host cell
Shigella actin to polymerize at one end of the bacterial cell. This forms an
“actin tail” that propels the bacterium within the cell. The force of
M cell the propulsion is so great that the bacterial cells are often driven
into neighboring cells. Listeria monocytogenes (causes meningitis)
does the same thing (figure 16.8). actin, p. 73 listeriosis, p. 648

Tissue 2 Shigella cells attach to the

base of the epithelial cells
Macrophages and induce these cells to
engulf them.

1 Macrophages in the Peyer’s patches engulf material that

passes through M cells. Shigella cells survive and
replicate, causing the phagocytes to undergo apoptosis.

FIGURE 16.7 Antigen-Sampling Processes Provide a

Mechanism for Invasion Shigella species use M cells to move across FIGURE 16.8 Actin Tail of Intracellular Listeria
the epithelial barrier. Once the bacterial cells are on the other side, monocytogenes Rapid polymerization of host cell actin (green) at
macrophages ingest them, but the bacteria are able to escape and one end of the bacterial cell (orange) propels the bacterium within
then infect other cells. the cell.
? What is the normal function of M cells? ? How does an actin tail benefit a bacterial cell?
 Part III Microorganisms and Humans 389

Avoiding Killing by Complement 1 Preventing Encounters with Phagocytes

System Proteins Some pathogens prevent phagocytosis by avoiding macrophages
As described in chapter 14, activation of the complement system and neutrophils altogether. The mechanisms include:
leads to three primary outcomes—lysis of foreign cells by mem- ■ C5a peptidase. This enzyme degrades the complement system
brane attack complexes (MACs), opsonization, and inflammation component C5a, a chemoattractant that recruits phagocytic
(see figure 14.10). Because the latter two outcomes are associated cells. Streptococcus pyogenes (causes strep throat) makes C5a
with phagocytosis, mechanisms that bacteria use to avoid them peptidase. C5a, p. 344 Streptococcus pyogenes, p. 487
will be discussed in the next section. Here, we will focus on how
■ Membrane-damaging toxins. These kill phagocytes and
bacteria avoid the lethal effects of MACs. Recall that Gram-
other cells, often by forming pores in their membranes.
negative bacteria are susceptible to MACs because the outer mem-
S. pyogenes makes a membrane-damaging toxin called strep-
brane serves as a target; MACs have little effect on Gram-positive
tolysin O. membrane-damaging toxins, p. 392
bacteria. complement system, p. 344
Bacteria that use mechanisms to avoid killing by the comple-
ment proteins are said to be serum resistant; strains of Neisseria 2 Avoiding Recognition and Attachment
gonorrhoeae that cause disseminated gonococcal infection are an
Some pathogens avoid being recognized by phagocytes. Recall
example. These strains hijack the mechanism that host cells use to
that phagocytes recognize and attach to foreign material more effi-
prevent their own surfaces from activating the complement system
ciently if opsonins such as C3b or antibodies coat it. Mechanisms
(see figure 14.12). By binding to the host’s complement regula-
that bacteria use to avoid opsonization include: opsonins, p. 347
tory proteins, they avoid complement activation by the alterna-
tive pathway, thereby postponing MAC formation. disseminated ■ Capsules. These have long been recognized for their abil-
gonococcal infection, p. 622 ity to prevent phagocytosis. Scientists now know that some
capsules do this by interfering with opsonization. In some
cases, they bind the host’s complement regulatory proteins
Avoiding Destruction by Phagocytes that inactivate C3b—a mechanism identical to that described
Phagocytosis involves multiple steps—including chemotaxis, earlier for serum-resistant bacteria. Rapid inactivation of
recognition and attachment, engulfment, and fusion of the phago- C3b prevents the molecule from being an effective opsonin,
some with lysosomes—that lead to the destruction of invading and it also avoids activation of the complement system by
microbes (see figure  14.10). Pathogens have evolved several the alternative pathway. Streptococcus pneumoniae (causes
mechanisms to avoid these destructive effects (figure 16.9). pneumonia) produces a capsule that does this.

FIGURE 16.9 Avoiding Destruction by Phagocytes

1 Prevent encounters
with phagocytes ? Mycobacterium tuberculosis appears to recruit phagocytes to the site of
C5a infection. Based on this information, which of the three methods shown
• C5a peptidase
• Cytolytic toxins here does the bacterium likely to use to avoid being destroyed?

Microbes

2 Avoid recognition
and attachment
• Capsules C3b Phagocyte
• M protein
• Fc receptors Lysosomes

Pseudopod Phagosome
C3b
Phagolysosome
C3b receptors
on phagocyte

Digestive
enzymes
3 Survive within phagocytes
• Escape from the phagosome
• Prevent phagosome-
lysosome fusion
• Survive within the phagosome
390 Chapter 16 Host-Microbe Interactions

■ M protein. This component of the cell wall of Streptococcus been ingested by a macrophage, it delays fusion of the phago-
pyogenes functions in a manner similar to that described some with the lysosome, allowing additional time for the microbe
for capsules. It binds a complement regulatory protein that to equip itself for growth within the phagolysosome.
inactivates C3b, thereby preventing it from being an effective
opsonin and avoiding the alternative pathway of complement Avoiding Antibodies
system activation. Pathogens that survive the innate defenses soon encounter an addi-
■ Fc receptors. These proteins bind the Fc region of antibod- tional obstacle, the adaptive defenses. For bacteria, the most impor-
ies, interfering with their function as opsonins (figure 16.10). tant of these are antibodies. Mechanisms for avoiding them include:
Recall that antibodies have two parts—the Fab region, which ■ IgA protease. This enzyme cleaves IgA, the class of antibody
binds specifically to antigens, and the Fc region, which func- found in mucus and other secretions. Neisseria gonorrhoeae
tions as a “red flag” (see figure 15.7). Bacterial cells that have and a variety of other pathogens produce IgA protease. This
Fc receptors coat themselves with antibody molecules with enzyme may also have other roles.
the Fab region projecting outward. The phagocytic cell has no
■ Antigenic variation. Some pathogens routinely alter the struc-
mechanism for recognizing the Fab regions, so this masks the
ture of their surface antigens. This allows them to stay ahead of
bacterial cell from phagocytes. Staphylococcus aureus and
antibody production by altering the very molecules antibodies
Streptococcus pyogenes make Fc receptors (protein A and
would otherwise recognize. Neisseria gonorrhoeae is able to
protein G, respectively). Fc and Fab regions, p. 360
vary the antigenic structure of its pili; antibodies produced by
3 Surviving Within Phagocytes the infected host in response to one variation of the pili cannot
bind effectively to another. antigenic variation, p. 178
Some bacteria make no attempt to avoid engulfment by phago-
cytes, instead using it as an opportunity. It allows them to hide ■ Mimicking host molecules. Pathogens sometimes cover them-
from antibodies, control some aspects of the immune response, selves with molecules similar to those normally found in the
and be transported to other locations in the body. Mechanisms host. This molecular mimicry takes advantage of the fact that
used to survive within phagocytes include: the immune system typically does not mount an attack against
“self” molecules. Certain strains of Streptococcus pyogenes have
■ Escape from the phagosome. Some pathogens escape from a capsule composed of hyaluronic acid, a polysaccharide found
the phagosome before it fuses with lysosomes. The bacte- in human tissues.
ria then multiply within the cytoplasm of the phagocyte,
protected from other host defenses. Listeria monocytogenes MicroAssessment 16.7
produces a molecule that forms pores in the phagosomal Serum-resistant bacteria avoid the killing effects of complement
membrane, allowing the bacterial cells to escape. Shigella system proteins. Mechanisms bacteria use to avoid destruction by
species lyse the phagosome before it fuses with lysosomes. phagocytes include preventing encounters with phagocytes, avoiding
■ Preventing phagosome-lysosome fusion. Bacteria that prevent recognition and attachment, and surviving within the phagocyte.
Mechanisms for avoiding antibodies include IgA protease, antigenic
phagosome-lysosome fusion avoid the otherwise inevitable expo-
variation, and mimicking host molecules.
sure to the destructive components of lysosomes. Salmonella spe-
cies can sense they have been ingested by a macrophage, and then 15. Describe how Fc receptors prevent phagocytosis.
respond by producing a protein that blocks the fusion process. 16. Describe three mechanisms pathogens may use to survive within
phagocytic cells.
■ Surviving within the phagolysosome. Relatively few microbes
17. Encapsulated organisms can be phagocytized once antibodies
can survive the destructive environment within the phagolyso- against the capsule have been produced. Why would this
some. Coxiella burnetii (causes Q fever), however, is able to be so? +
withstand the conditions. It appears that once the organism has

Bacterium Fc receptor on
bacterium (binds
Fab region of the antibody the Fc region of
(binds to antigen) an antibody)

Fc region of the antibody

(phagocytes recognize
Antibody and bind this region as
an initial step in phagocytosis)
(a) (b)
FIGURE 16.10 Fc Receptors Foil Opsonization by Antibodies (a) The normal orientation of antibody molecules on the surface of a bacte-
rium; note that the Fc region of the antibody projects from the bacterial cell, making it available for a phagocyte to recognize and bind. (b) The
effect of Fc receptors on a bacterial cell’s surface; the receptors bind the Fc portion of antibodies, regardless of their specificity.
? Would antibodies that the Fc receptors bind be specific for the bacterium that makes the receptors? Why or why not?
 Part III Microorganisms and Humans 391

16.8 ■ Damage to the Host Exotoxins

A number of Gram-positive and Gram-negative pathogens pro-
Learning Outcomes duce exotoxins—proteins that have very specific damaging
11. Describe the difference between exotoxins and endotoxins. effects (table 16.1). Exotoxins are often a major cause of damage
12. Compare and contrast neurotoxins, enterotoxins, and cytotoxins, to an infected host.
giving two examples of each. Exotoxins are either secreted by the bacterium or leak into
13. Explain how inflammation and antibodies can cause damage. the surrounding fluid following lysis of the bacterial cell. In most
cases, the pathogen must colonize a body surface or tissue to
Damage due to infection can be the result of direct effects of the produce enough toxin to cause damage. With foodborne intoxica-
pathogen, such as toxins produced, or indirect effects, such as the tion, however, the bacterial cells multiply in a food product where
immune response. In many cases, the damage helps the organism they produce toxin that is then consumed. In the case of botulism,
exit the host, allowing it to spread to others. For example, Vibrio ingestion of even tiny amounts of botulinum toxin is sufficient to
cholerae (causes cholera) induces watery diarrhea—up to 20 liters cause paralysis. Like most other exotoxins, botulinum toxin can
of microbe-containing fluid in one day! In areas of the world be destroyed by heating. botulism, p. 652
without adequate sewage treatment, this can lead to contaminated Exotoxins can act locally, or they may be carried in the
water supplies and widespread outbreaks. Bordetella pertussis bloodstream throughout the body, causing systemic effects.
(causes whooping cough) triggers severe bursts of coughing, pro- Corynebacterium diphtheriae (causes diphtheria) grows and releases
pelling the respiratory pathogens into the air. its exotoxin in the throat. There, the toxin destroys local cells, leading

TABLE 16.1 Exotoxins Produced by Various Primary Pathogens

Name of Disease; Characteristics Page
Example Name of Toxin of the Disease Mechanism Reference

A-B TOXINS—Composed of two subunits, A and B. The A subunit is the toxic, or active, part; the B subunit binds to the target cell.
Neurotoxins
Clostridium Botulism; botulinum toxin Flaccid paralysis Blocks transmission of nerve signals to p. 652
botulinum the muscles by preventing the release of
acetylcholine.
Clostridium tetani Tetanus; tetanospasmin Spastic paralysis Blocks the action of inhibitory neurons by p. 555
preventing the release of neurotransmitters.

Enterotoxins
Enterotoxigenic Traveler’s diarrhea; heat- Severe watery diarrhea Modifies a regulatory protein in intestinal p. 590
E. coli labile enterotoxin (cholera- cells, causing those cells to continuously
like toxin) secrete electrolytes and water.
Vibrio cholerae Cholera; cholera toxin Severe watery diarrhea Modifies a regulatory protein in intestinal p. 586
cells, causing those cells to continuously
secrete electrolytes and water.

Cytotoxins
Bacillus anthracis Anthrax; edema factor, Inhaled form—septic shock; Edema factor modifies a regulatory protein p. 497
lethal factor cutaneous form—skin in cells, causing accumulation of fluids.
lesions Lethal factor inactivates proteins involved in
cell signaling functions.
Bordetella pertussis Pertussis (whooping Sudden bouts of violent Modifies a regulatory protein in respiratory p. 501
cough); pertussis toxin coughing cells, causing accumulation of respiratory
secretions and mucus. Other factors also
contribute to the symptoms.
Corynebacterium Diphtheria; diphtheria Pseudomembrane in the Inhibits protein synthesis by inactivating an p. 490
diphtheriae toxin throat; heart, nervous elongation factor of eukaryotic cells. Kills
system, kidney damage local cells (in the throat) and is carried in the
bloodstream to various organs.
E. coli O157:H7 Bloody diarrhea, hemolytic Diarrhea that may be Inactivates the 60S subunit of eukaryotic p. 590
uremic syndrome; shiga bloody; kidney damage ribosomes, halting protein synthesis.
toxin
Shigella Dysentery, hemolytic Diarrhea that contains Inactivates the 60S subunit of eukaryotic p. 588
dysenteriae uremic syndrome; shiga blood, pus, and mucus; ribosomes, halting protein synthesis.
toxin kidney damage

(continued)
392 Chapter 16 Host-Microbe Interactions

TABLE 16.1 Exotoxins Produced by Various Primary Pathogens (Continued)

Name of Disease; Characteristics Page
Toxins Name of Toxin of the Disease Mechanism Reference
MEMBRANE-DAMAGING TOXINS (cytotoxins)—Disrupt plasma membranes.
Clostridium Gas gangrene; α-toxin Extensive tissue damage Removes the polar head group on the p. 558
perfringens phospholipids in the membrane, destroying
membrane integrity.
Staphylococcus Wound and other Accumulation of pus Inserts into membranes, forming pores that p. 551
aureus infections; leukocidin allow fluids to enter the cells.
Streptococcus Pharyngitis and other Accumulation of pus Inserts into membranes, forming pores that p. 487
pyogenes infections; streptolysin O allow fluids to enter the cells.
SUPERANTIGENS—Override the specificity of the T-cell response.
Staphylococcus Foodborne intoxication; Nausea and vomiting Not well understood with respect to how p. 757
aureus (certain staphylococcal the ingested toxins lead to the characteristic
strains) enterotoxins symptoms of foodborne intoxication.
Staphylococcus Staphylococcal toxic shock; Fever, vomiting, diarrhea, Systemic toxic effects due to the resulting p. 619
aureus (certain toxic shock syndrome toxin muscle aches, rash, low massive release of cytokines.
strains) (TSST) blood pressure
Streptococcus Streptococcal toxic shock; Fever, vomiting, diarrhea, Systemic toxic effects due to the resulting p. 553
pyogenes (certain streptococcal pyrogenic muscle aches, rash, low massive release of cytokines.
strains) exotoxins (SPE) blood pressure
OTHER TOXIC PROTEINS
Staphylococcus Scalded-skin syndrome; Separation of the outer Thought to break ester bonds that hold the p. 527
aureus exfoliatin layer of skin layers of skin together.
Various organisms Various diseases; proteases, Tissue damage Degrades proteins, lipids, and other
lipases, and other hydrolases compounds that make up tissues.

to the accumulation of dead host cells, pus, and blood. This forms a MicroByte
membrane that sometimes dislodges and blocks the airway. The toxin Botox is a dilute suspension of botulinum toxin (see Perspective 31.1).
can be absorbed and carried to the heart, nervous system, and other
organs, causing additional damage. diphtheria, p. 490 A-B Toxins
Because exotoxins are proteins, the immune system can gener- A-B toxins consist of two parts: the A subunit is the toxic (active)
ally produce neutralizing antibodies (see figure 15.8). Unfortunately, portion and the B subunit binds to a specific surface molecule on cells
many exotoxins are so powerful that fatal damage occurs before an (figure 16.11). In other words, the A subunit, usually an enzyme, is
adequate immune response is mounted. This is why vaccination is responsible for the effects of the toxin on a cell, whereas the B subunit
so important. It prevents otherwise common and often fatal dis- dictates the type of cell to which the toxin is delivered.
eases such as tetanus and diphtheria (see table 18.1). The vaccines The structure of A-B toxins offers novel approaches for the
against tetanus and diphtheria are toxoids, which are inactivated development of vaccines and therapies. For example, a fusion pro-
toxins. A vaccine against botulinum toxin is also available, but it is tein that contains diphtheria toxin is used to treat a type of cancer
not part of routine vaccination because the risks of developing the called cutaneous T-cell lymphoma. By fusing the toxin to a cyto-
disease are extremely low if sensible food preparation procedures kine that binds T cells, the toxin is delivered to those cells, includ-
are followed. If a person develops symptoms of a toxin-mediated ing cancerous ones. In some countries, the B subunit of cholera
disease, he or she can be treated with antitoxin, a suspension of toxin is used as an orally administered vaccine against cholera.
neutralizing antibodies. vaccines, p. 421 antitoxin, p. 420 Antibodies that bind the B subunit prevent cholera toxin from
Many exotoxins can be grouped into functional categories attaching to intestinal cells, thus protecting the vaccine recipient.
according to the tissues they affect (table  16.1). Neurotoxins Researchers are now experimenting with joining medically useful
damage the nervous system, causing symptoms such as paralysis. compounds to B subunits, allowing medications to be delivered
Enterotoxins cause symptoms associated with intestinal disturbance, specifically to the cell type targeted by the B subunit.
such as diarrhea and vomiting. Cytotoxins damage a variety of dif-
ferent cell types, either by interfering with essential cellular mecha- Membrane-Damaging Toxins
nisms or by lysing cells. Some exotoxins do not fall into any of these Membrane-damaging toxins are cytotoxins that disrupt plasma
groups, instead causing symptoms associated with excessive stimula- membranes, causing the cell to lyse. Many lyse red blood cells,
tion of the immune response. Most exotoxins fall into three general causing hemolysis that can be observed when the organisms are
categories that reflect their structure and general mechanism of grown on blood agar; because of this, many of these toxins are
action: A-B toxins, membrane-damaging toxins, and superantigens. referred to as hemolysins. hemolysis, p. 96 blood agar, p. 95
Active subunit Part III Microorganisms and Humans 393
A
Binding subunit B Some membrane-damaging toxins insert themselves into
membranes, forming pores that allow fluids to enter the cell. One
pore-forming toxin is streptolysin O, the compound responsible
for the characteristic β-hemolysis of Streptococcus pyogenes
Binding site grown anaerobically on blood agar (see figure 4.10). Recall that
streptolysin O also helps S. pyogenes avoid phagocytosis.
Phospholipases hydrolyze phospholipids in the plasma
membrane. The α-toxin of Clostridium perfringens (causes
1 B subunit binds to a
specific molecule 2 Toxin is taken up gas gangrene) is an example. phospholipid, p. 35 gas gangrene,
on the host cell. by endocytosis. p. 558

Superantigens
3 Toxin subunits separate
Superantigens override the specificity of the helper T-cell
allowing the A subunit response, causing toxic effects due to the massive release of
to enter the cytoplasm. cytokines by TH cells (effector helper T cells). Superantigens
include toxic shock syndrome toxin (TSST) as well as several other
toxins produced by Staphylococcus aureus and Streptococcus
FIGURE 16.11 The Action of A-B Toxins The A subunit pyogenes. helper T cells, p. 360
is toxic and the B subunit binds to specific receptors on cells. After Superantigens short-circuit the normal specificity of antigen
being take up by endocytosis, the A and B subunits separate in the
recognition by a helper T cell. They do this by binding simultane-
acidic endosome. The A subunit then enters the cell’s cytoplasm and
exerts its toxic effect. ously to the outer portion of the major histocompatibility (MHC)
class II molecule on antigen-presenting cells and the T-cell receptor
? Would an antibody response against the B subunit protect against (figure  16.12). The T-cell machinery interprets the binding to
the effects of the toxin?
mean that the T-cell receptor recognized the antigen presented
on the MHC molecule, when it prob-
Antigen-presenting cell Antigen-presenting cell ably did not. Whereas an antigen usually
stimulates about one in 10,000 helper T
cells, superantigens stimulate as many
as one in five. This results in a massive
release of cytokines, leading to fever,
nausea, vomiting, and diarrhea. Shock
may occur, with organ failure, circula-
tory collapse, and even death. In addi-
MHC class II tion, the immune response is suppressed
molecule
because many T cells undergo apoptosis
following the stimulation. Superantigens
are also suspected of contributing to
Peptide Peptide not autoimmune diseases. By overriding the
recognized by recognized by
T-cell receptor T-cell receptor
normal control mechanisms of adaptive
immunity, superantigens may promote
proliferation of T cells that respond
Superantigen to healthy “self.” MHC class II molecules,
T-cell
receptor p. 369 apoptosis, p. 350
The exotoxins produced by
Staphylococcus aureus strains that cause
foodborne intoxication are superantigens.
Helper T cell Helper T cell Although they cause nausea and vomiting
and are therefore referred to as enterotoxins,
their structure and action are very different
from the enterotoxins of Vibrio cholerae
a Helper T cell that recognizes peptide is b Helper T cell that does not recognize and E. coli strains. The mechanism by which
activated; it proliferates and releases peptide is activated because of superantigen;
cytokines. it proliferates and releases cytokines. they induce vomiting is poorly understood.
Unlike most other exotoxins, the entero-
toxins produced by Staphylococcus aureus
FIGURE 16.12 Superantigens (a) Normal antigen presentation. (b) Superantigens are heat-stable. Even thorough cooking of
short-circuit the normal specificity of the helper T-cell response by binding simultaneously to the foods contaminated with these toxins will
outer portion of the MHC class II molecule and the T-cell receptor. not prevent illness. Staphylococcus aureus
? What specifically causes the toxic effects of superantigens? food poisoning, p. 757
394 Chapter 16 Host-Microbe Interactions

Other Toxic Proteins activation of the coagulation cascade. The overwhelming systemic
Various proteins that are not A-B toxins, superantigens, or response causes a dramatic drop in blood pressure, disseminated
membrane-damaging toxins can have damaging effects. An impor- intravascular coagulation (DIC), and fever (see figure 27.3). This
tant example is the toxin produced by strains of Staphylococcus array of symptoms associated with systemic bacterial infection is
aureus that cause scalded skin syndrome. This toxin, exfoliatin, called septic shock; when it is caused by endotoxin, it may also be
destroys material that binds together the layers of skin, causing the called endotoxic shock. DIC, p. 674 septic shock, p. 674

outer layer to separate (see figure 22.4). The bacteria might be grow- Lipid A is embedded in the Gram-negative outer mem-
ing in a small, localized lesion but the toxin spreads systemically. brane and does not cause a response unless it is released. This
Various hydrolytic enzymes including proteases, lipases, and occurs primarily when a bacterium lyses, which can happen as
collagenases break down tissue components. Along with destroy- a result of phagocytosis, activation of the complement system
ing tissues, some of these enzymes help the bacteria spread. (due to membrane attack complexes), and treatment with cer-
tain types of antibiotics. Once released from a cell, the LPS
molecules can activate the innate and adaptive defenses by a
Endotoxin and Other Bacterial variety of mechanisms. Monocytes, macrophages, and other
Cell Wall Components cells have toll-like receptors (TLRs) that detect liberated LPS,
The host defenses are primed to respond to various bacterial cell inducing the cells to produce pro-inflammatory cytokines.
wall components, including lipopolysaccharide and peptidogly- LPS also functions as a T-independent antigen; at high con-
can. A strong and widespread immune response to these com- centrations it activates a variety of different B cells, regard-
pounds, however, can have toxic effects. less of the specificity of their B-cell receptor. TLRs, p.  342
T-independent antigens, p. 367
Endotoxin Endotoxin is heat-stable; it is not destroyed by autoclav-
Endotoxin is lipopolysaccharide (LPS), the molecule that makes ing. Consequently, solutions intended for intravenous (IV)
up the outer layer of the outer membrane of the Gram-negative administration must not only be sterile, but free of endotoxin as
cell wall (see figure  3.33). The name is somewhat unfortunate, well. Disastrous results including death have resulted from IV
because it implies that endotoxin is “inside the cell,” and, con- fluids contaminated with endotoxin. To verify that fluids are not
versely, that exotoxins are “outside the cell.” This is misleading, contaminated, a very sensitive test known as the Limulus amoe-
because endotoxin is an integral part of the outer membrane, bocyte lysate (LAL) assay is done. This uses proteins extracted
whereas exotoxins are proteins that may or may not be secreted from blood of the horseshoe crab (Limulus polyphemus) that
by the bacterial cell. Unlike most exotoxins, endotoxin cannot be form a gel-like clot when exposed to endotoxin; as little as 10 to
converted to an effective toxoid for immunization. Table  16.2 20 picograms (1 picogram = 10–12 grams) of endotoxin per mil-
summarizes some of the other differences between exotoxins and liliter can be detected using the LAL. Horseshoe crabs are one
endotoxin. lipopolysaccharide, p. 60 of this planet’s more unique and ancient life-forms. The critical
Recall from chapter 3 that the lipopolysaccharide mol- role they play in this test has led to an increased awareness of the
ecule contains lipid A. This component triggers an inflammatory importance of their habitat. A non-lethal system of capture, blood
response and is responsible for the adverse effects of LPS. When sampling, and release has been developed.
lipid A is present in a localized region, the response helps clear
an infection. It is a different situation entirely, however, when the Other Bacterial Cell Wall Components
infection is systemic, as in a bloodstream infection. Imagine a state Peptidoglycan and other bacterial cell wall components can cause
in which inflammation occurs throughout the body—extensive symptoms similar to those that characterize the response to endo-
leakage of fluids from permeable blood vessels and widespread toxin. The systemic response leads to septic shock.

TABLE 16.2 Comparison of Exotoxins and Endotoxin

Property Exotoxins Endotoxin

Bacterial source Gram-positive and Gram-negative species Gram-negative species only

Location in the bacterium Synthesized in the cytoplasm; may or may Component of the outer membrane of the Gram-negative cell wall
not be secreted
Chemical nature Protein Lipopolysaccharide (the lipid A component)
Ability to form a toxoid Generally No
Heat stability Generally inactivated by heat Heat-stable
Mechanism A distinct toxic mechanism for each Innate immune response; a systemic response leads to fever, a dramatic
drop in blood pressure, and disseminated intravascular coagulation
Toxicity Generally very potent; some are among Small amounts in a localized area lead to an appropriate immune
the most potent toxins known. response that helps clear an infection, but systemic distribution can
be deadly
 Part III Microorganisms and Humans 395

Damaging Effects of 16.9 ■ Mechanisms of Viral

the Immune Response Pathogenesis
Although the immune response eliminates invading microbes, it
can inadvertently damage host tissues as well. The reactions to Learning Outcomes
endotoxin and other cell wall components are examples of damag- 14. Describe how viruses bind to host cells, and how they spread to
ing effects of the immune response, but are typically considered a other cells.
toxic effect of the bacterium because the reactions can be imme- 15. Describe how viruses avoid interferon, regulate apoptosis, and
diate and overwhelming. The damaging responses discussed next avoid antibodies.
become apparent more slowly.
To infect a host, a virus must enter an appropriate cell, use the
Damage Associated with Inflammation cell’s machinery for replication, keep the host from recognizing
The inflammatory response itself can destroy tissue because and destroying the infected cell, and then move to new cells or
phagocytic cells recruited to the area release some of the enzymes hosts. In carrying out these functions, many viruses damage host
and toxic products they contain. The life-threatening aspects of cells and induce inflammatory responses, causing disease.
bacterial meningitis, for example, are due to the inflammatory
response itself. Complications of certain sexually transmitted
Binding to Host Cells and Invasion
infections are also due to the damage associated with inflam-
mation. For example, if Neisseria gonorrhoeae or Chlamydia As discussed in chapter 13, viruses attach to target cells via spe-
trachomatis infections involve the fallopian tubes, the inflammatory cific receptors. Only cells that bear the receptor can be infected, so
response can lead to scarring that obstructs the tubes, either prevent- this influences the host range and tissue specificity of a particular
ing fertilization or predisposing a woman to an ectopic pregnancy virus. For example, HIV’s receptor is CD4, a molecule found on
(meaning the fertilized egg implants outside of the uterus). helper T cells and macrophages. Picornaviruses and reoviruses
bind to receptors on M cells in the Peyer’s patches of the intesti-
Damage Associated with Adaptive Immunity nal tract. Once attached, viruses then enter the cells using either
The adaptive immune response can also lead to damaging effects. receptor-mediated endocytosis or fusion with the plasma mem-
Mechanisms include: brane (see figure 13.13). CD4, p. 369
Virions released from a cell can either infect neighboring
■ Immune complexes. When antibodies bind to antigens, the cells or disseminate via the bloodstream or lymphatic system
complexes can settle in the kidneys and joints, where they to other tissues. Polioviruses, for example, initially infect
activate the complement system, causing destructive inflam- cells in the throat and intestinal tract. Upon release from these
mation. An example is acute glomerulonephritis, a compli- cells, the virus enters the bloodstream and may then spread to
cation that can follow skin and throat infections caused by infect motor nerve cells of the brain and spinal cord, causing
Streptococcus pyogenes; the immune complexes that form paralysis. polio, p. 656
trigger a response that damages kidney structures called
glomeruli. acute glomerulonephritis, p. 490

■ Cross-reactive antibodies. Certain antibodies produced in

Avoiding Immune Responses
response to an infection bind to the body’s own tissues, pro- Viruses must avoid the host defenses that detect and eliminate
moting an autoimmune response. Acute rheumatic fever—a invaders. These include interferon, apoptosis, and antibodies.
complication that can follow strep throat—is thought to be interferon, p. 341 apoptosis, p. 350
due to antibodies against S. pyogenes binding to normal tissue
proteins. This occurs most frequently in people with certain Avoiding the Antiviral Effects of Interferons
MHC types (see Perspective 15.1). acute rheumatic fever, p. 489 Early in viral infection, interferons play an important role in lim-
iting viral spread (see figure  14.9). They cause cells to produce
enzymes that, when activated, prevent viral replication. To avoid
MicroAssessment 16.8
this, some viruses encode proteins that shut down expression of
Damage can be due to exotoxins, including A-B toxins, superantigens, host genes. Others avoid interferon’s effects by interfering with
membrane-damaging toxins, and other toxic proteins. Endotoxin and activation of the enzymes.
other cell wall components in the bloodstream can result in septic
shock. The inflammatory response can cause tissue damage that leads Regulating Host Cell Death
to scarring. Immune complexes can trigger damaging inflammation
in the kidneys and joints; cross-reactive antibodies can lead to an Many viruses regulate host cell death. Some kill the host cell
autoimmune response. once it has produced virions, and others prevent the host cell from
18. What is an A-B toxin? dying prematurely.
Recall that the interferon response induces apoptosis in virally
19. How is an enterotoxin different from endotoxin?
infected cells. Some viruses prevent this from happening, thereby
20. Home-canned foods should be boiled before consumption to
giving the virus more time to replicate. They do this by controlling
prevent botulism. Considering that this treatment does not
destroy endospores, why would it prevent the disease? + a protein called p53 that regulates apoptosis in cells. When this
protein is inhibited, tumors sometimes develop. Papillomaviruses,
396 Chapter 16 Host-Microbe Interactions

a group of viruses that cause various types of warts, interfere with fever, a severe illness caused by dengue viruses, is associated with
the normal function of p53. papillomaviruses, p. 308 antibody-dependent enhancement of infection. dengue hemorrhagic
To avoid being detected by the cellular immune response, fever, p. 683
many types of viruses interfere with antigen presentation by MHC
class I molecules. Herpesviruses, for example, block the move- MicroAssessment 16.9
ment of the molecules to the surface of the infected cell, so that
Proteins on the surface of virus particles function as adherence
TC cells (effector cytotoxic T cells) cannot inspect the proteins
factors. Viruses, as a group, have several mechanisms for avoiding
being made. The immune system, however, is prepared for such the effects of interferon, regulating host cell death, and avoiding
a strategy. Natural killer (NK) cells recognize stressed cells that antibodies.
lack MHC class I molecules and destroy them (see figure 15.23). 21. From a virus’s perspective, why would it be beneficial to prevent
Perhaps not surprisingly, some viruses have methods to trick NK apoptosis?
cells. Cytomegalovirus (CMV), which causes disease in immuno- 22. How do cytomegaloviruses avoid the cellular immune response?
compromised people, encodes production of “fake” MHC class I
23. Why would various unrelated viral infections often include a
molecules. These decoy versions are displayed on the surface of similar set of symptoms (fever, headache, fatigue, and runny
the host cell, tricking the immune system into believing that all is nose)? +
well in the cell (figure 16.13). antigen presentation by MHC class I,
p. 369 natural killer cells, p. 374 cytomegaloviruses, p. 711

Avoiding Antibodies
16.10 ■ Mechanisms of Eukaryotic
Antibodies generally control the spread of viruses by neutralizing Pathogenesis
extracellular virions (see figure 15.8). To avoid antibodies, some
Learning Outcomes
viruses move directly from one cell to its immediate neighbors.
Other viruses remain intracellular by forcing cellular neighbors 16. Describe the mechanisms of pathogenesis of dermatophytes,
Candida albicans, and the dimorphic fungi.
to fuse, forming a syncytium. HIV induces syncytia formation.
syncytium, p. 326
17. Compare and contrast the mechanisms of pathogenesis of
protozoa and helminths.
The surface antigens of some viruses change rapidly, outpac-
ing the body’s capacity to produce effective neutralizing antibodies.
Pathogenesis of eukaryotic cells, including fungi and protozoa,
This rapid evolution occurs because the replicases of RNA viruses
involves the same basic scheme as that of bacterial pathogens—
and the reverse transcriptase of HIV have no proofreading abil-
colonization, evasion of host defenses, and damage to the host.
ity, so mutations occur fairly often. Thus, as the viruses replicate,
The mechanisms, however, are generally not as well understood.
they give rise to pools of genetically altered virions. Eventually,
an immune response against essential proteins that cannot tolerate
extensive mutations may eventually clear the viruses. This is not Fungi
the case for HIV, however. An HIV-infected person produces many Most fungi, such as yeasts and molds, are saprophytes, meaning
antibodies against HIV, but none are effective in clearing the virus. they acquire nutrients from dead and dying material; those that
replicase, p. 320 reverse transcriptase, p. 320 can cause disease are generally opportunists, although notable
Some viruses actually take advantage of antibodies, using them exceptions exist.
to help enter macrophages. Opsonized viral particles are engulfed Members of a group of fungi referred to as dermatophytes
by phagocytes, which they then infect. Dengue hemorrhagic cause superficial infections of hair, skin, and nails, but do not

1 Virus 2 Fake MHC 3 4

Peptide from Tc cell
normal protein class I molecule

Viral
MHC class I genome
molecule

NK
cell

Virus infects cell. Viral genome directs the cell Because of the fake MHC class I Infected cell survives and
to make fake MHC class I molecules, neither Tc cells nor carries the viral genome.
molecules that cannot NK cells can recognize that the
present peptides from cell is infected.
cytoplasmic proteins.

FIGURE 16.13 The Role of MHC Class I Decoys in Viral Pathogenesis

? What is the role of MHC class I molecules in adaptive immunity?
 Part III Microorganisms and Humans 397

invade deeper tissues. These fungi have keratinase enzymes that by this species. Giardia lamblia uses a disc that functions as a suc-
break down the keratin in superficial tissues, allowing the fungi tion cup to attach to the intestinal surface; it also has an adhesin
to use this protein as a source of nutrition. Dermatophytes cause associated with the disc that helps the initial attachment.
diseases such as ringworm and athlete’s foot. Protozoa and helminths use a variety of mechanisms to avoid
Fungi in the normal microbiota, especially the yeast Candida antibodies. Some hide within cells, thus avoiding exposure to
albicans, can cause disease in immunocompromised hosts. Factors antibodies as well as certain other defenses. Malarial parasites,
that can lead to excessive growth of C. albicans include AIDS, for example, produce enzymes that allow them to penetrate red
uncontrolled diabetes, severe burns, and inhibition of normal blood cells. This cell type does not present antigen to TC cells,
microbiota due to hormonal influences or antibiotic treatment. so the parasite escapes antibodies as well as the cell-mediated
C. albicans generally infects the mucous membranes, causing immune defenses. Leishmania species survive and multiply within
thrush (an infection of the throat and mouth) or vulvovaginitis. macrophages when phagocytized. Parasites such as the African
The most serious fungal infections are caused by dimor- trypanosomes, the cause of sleeping sickness, prevent antibodies
phic fungi. These occur as molds in the environment, and from recognizing them by routinely varying their surface antigens
when the small airborne conidia are inhaled, they lodge deep through antigenic variation. Schistosoma species coat themselves
within the lungs. There, they develop into other forms, usually with host proteins, thereby disguising themselves. Some parasitic
yeasts. The immune system generally controls the infection, so worms appear to suppress immune responses in general.
most cases are asymptomatic. The situation is entirely differ- The extent and type of damage caused by parasites varies
ent if the person is immunocompromised, however, because tremendously. In some cases, the parasites compete for nutrients
the infections can become life-threatening. dimorphic fungi, in the intestinal tract, contributing to malnutrition of the host.
p. 285 conidia, p. 287 Helminths may accumulate in high enough numbers or grow long
Some fungi produce toxins, collectively referred to as myco- enough to block the intestines or other organs. Some parasites
toxins. Aspergillus flavus, for example, a fungus that grows on produce enzymes that digest host tissue, causing direct damage.
certain grains and nuts, produces aflatoxin. Ingestion of this toxin In other cases, damage is due to the immune response; examples
can damage the liver and increases the risk of liver cancer. Fungal include the high fevers that characterize malaria, and the granulo-
spores and other products can cause hypersensitivities in some matous response to Schistosoma eggs.
people. hypersensitivities, p. 402 
MicroAssessment 16.10
Protozoa and Helminths Pathogenic mechanisms of fungi, protozoa, and helminths are not as
well understood as those of bacteria; however, they involve the same
Most protozoa and helminths either live within the intestinal tract basic scheme—colonization, evasion of host defenses, and damage to
or enter the body via the bite of an arthropod. Schistosoma species, the host.
however, can enter the skin directly (see Perspective 14.1). 24. What is the importance of keratinase?
Like bacteria and viruses, eukaryotic parasites attach to host 25. How do the African trypanosomes avoid the effects of
cells via specific receptors. Plasmodium vivax, one of the two antibodies?
most common causes of malaria, attaches to the Duffy blood 26. Why would relatively few people of West African ancestry
group antigen on red blood cells. Most people of West African have the Duffy blood group antigen? +
ancestry lack this antigen and are therefore resistant to infection

FUTURE CHALLENGES 16.1

The Potential of Probiotics
Considering the important protective roles the Marketing claims touting probiotics can shorten the duration of certain diarrheal ill-
normal microbiota play in human health, it is abound, but there are still many unanswered nesses. In contrast, studies that used a mixture
not surprising that administering live benefi- questions. For example, will a microbe that has containing L. bulgaricus and L. acidophilus
cial microbes, referred to as probiotics, has beneficial effects in vitro be stable in a food generally showed no helpful effect. Because of
been suggested as therapy for diarrheal and or supplement so that an adequate dose can be this, products that list ingredients such as “live
other diseases. In vitro studies certainly sup- consumed? And even if it is stable in the pack- active culture” or “lactic acid bacteria” do not
port their use, indicating that some strains of age, will it survive passage through the stomach necessarily contain a beneficial strain.
bacteria interfere with the growth or toxin pro- so that viable cells enter the intestinal tract? If it Complicating matters is that probiotics
duction of certain pathogens. Animal studies does access the intestines, can it colonize there, are considered “dietary supplements,” so there
using probiotics have also shown promising or will it simply pass through as part of feces? is little regulatory control over health claims.
results. For example, chicks fed a mixture of Beneficial effects of probiotics observed Early studies such as those using L. rhamnosus
Lactobacillus species that normally inhabit in humans so far are species and strain spe- GG certainly highlight their potential, but larger
poultry intestines were less likely to be subse- cific. One species may be protective, whereas and well-controlled scientific studies of probiot-
quently colonized by the pathogen Salmonella another closely related organism is not. For ics are needed to provide more data as to their
enterica. But which probiotics will be success- example, several small studies indicate that effectiveness in curing and preventing various
ful in treating humans? consumption of Lactobacillus rhamnosus GG medical conditions.
398 Chapter 16 Host-Microbe Interactions

Summary
MICROBES, HEALTH, AND DISEASE Delivering Effector Proteins to Host Cells
Type III secretion systems of Gram-negative bacteria allow them to
16.1 ■ The Anatomical Barriers as Ecosystems
deliver proteins directly to host cells (figure 16.5).
In mutualism, both partners benefit, in commensalism, one partner
benefits while the other is unaffected, and in parasitism, the parasite ■
16.6 Invasion—Breaching the Anatomical Barriers
benefits at the expense of the host.
Penetrating the Skin
16.2 ■ The Normal Microbiota (figure 16.1) Bacteria take advantage of trauma that destroys the integrity of the skin
or rely on arthropods to inject them.
The Protective Role of the Normal Microbiota
The normal microbiota excludes pathogens by covering binding sites Penetrating Mucous Membranes
that might otherwise be used for attachment, consuming available nutri- Some pathogens induce mucosal epithelial cells to engulf bacterial
ents, and producing compounds toxic to other bacteria. The normal flora cells; others exploit antigen-sampling processes (figures 16.6, 16.7).
primes the adaptive immune system.
16.7 ■ Avoiding the Host Defenses
The Dynamic Nature of the Normal Microbiota
The composition of the normal microbiota continually changes in Hiding Within a Host Cell
response to host factors including hormonal changes, and type and Some bacteria can evade the innate defenses, as well as some aspects of
quantity of food consumed. the adaptive defenses, by remaining inside host cells.
Avoiding Killing by Complement System Proteins
16.3 ■ Principles of Infectious Disease
Some serum-resistant bacteria postpone the formation of membrane
Infectious diseases have characteristic signs and symptoms. A
attack complexes by interfering with activation of the complement sys-
secondary infection can occur as the result of a primary infection.
tem via the alternative pathway.
Pathogenicity
Avoiding Destruction by Phagocytes (figure 16.9)
A primary pathogen causes disease in otherwise healthy individuals;
Mechanisms to prevent encounters with phagocytes include C5a pepti-
an opportunistic pathogen causes disease only when the body’s innate
dase and membrane-damaging toxins. Mechanisms to avoid recognition
or adaptive defenses are compromised. Virulence refers to the degree
and attachment by phagocytes include capsules, M protein, and Fc
of pathogenicity of an organism.
receptors (figure  16.10). Mechanisms to survive within the phagocyte
Characteristics of Infectious Disease include escape from the phagosome, preventing phagosome-lysosome
Communicable or contagious diseases spread from one host to fusion, and surviving within the phagolysosome.
another; ease of spread partly reflects the infectious dose. Stages
Avoiding Antibodies
of infectious disease include the incubation period, illness, and
convalescence; during the illness a person experiences signs and symp- Mechanisms to avoid antibodies include IgA protease, antigenic
toms of the disease (figure 16.2). Infections can be described as acute, variation, and mimicking “self.”
chronic, or latent, depending on the timing and duration of symptoms.
16.8 ■ Damage to the Host
Infections can be localized or systemic.
Exotoxins (table 16.1)
16.4 ■ Establishing the Cause of Infectious Disease Exotoxins are proteins that have very specific damaging effects; they
Koch’s Postulates may act locally or cause dramatic systemic effects. Many can be grouped
into categories such as neurotoxins, enterotoxins, or cytotoxins. The
Koch’s postulates are used to establish the cause of infectious disease
toxic activity of A-B toxins is mediated by the A subunit; binding to
(figure 16.3).
specific cells is mediated by the B subunit (figure  16.11). Membrane-
Molecular Koch’s Postulates damaging toxins disrupt cell membranes either by forming pores or
Molecular Koch’s postulates are used to identify virulence factors that by removing the polar head group on phospholipids in the membrane.
contribute to disease. Superantigens override the specificity of the T-cell response, causing
systemic effects due to the massive release of cytokines (figure 16.12).
MECHANISMS OF PATHOGENESIS Endotoxin and Other Bacterial Cell Wall Components

■
The symptoms associated with endotoxin are due to a vigorous host
16.5 Establishing Infection
response. Lipid A of lipopolysaccharide is responsible for its toxic
Adherence properties. Peptidoglycan and certain other components induce various
Bacteria use adhesins to bind to host cells (figure 16.4). cells to produce pro-inflammatory cytokines.

Colonization Damaging Effects of the Immune Response

Rapid turnover of pili, antigenic variation, and IgA proteases allow The release of enzymes and toxic products from phagocytic cells can
bacteria to avoid the effects of secretory IgA. Siderophores enable damage tissues. Immune complexes can cause kidney and joint dam-
microbes to scavenge iron. age; cross-reactive antibodies can result in an autoimmune response.
 Part III Microorganisms and Humans 399

16.9 ■ Mechanisms of Viral Pathogenesis 16.10 ■ Mechanisms of Eukaryotic Pathogenesis

Binding to Host Cells and Invasion Fungi
Viruses attach to specific receptors on the target cell. Saprophytes are generally opportunists; dermatophytes cause superfi-
cial infections of skin, hair, and nails. The most serious fungal infec-
Avoiding Immune Responses
tions are caused by dimorphic fungi.
Some viruses can avoid the effects of interferon; some can regu-
late apoptosis of the host cell (figure  16.13). To subvert the role of Protozoa and Helminths
antibodies, some viruses transfer directly from cell to cell; the surface Eukaryotic parasites attach to host cells via specific receptors. They
antigens of some viruses change quickly, outpacing the production of use a variety of mechanisms to avoid antibodies; the extent and type of
antibodies. damage they cause varies tremendously.

Review Questions
Short Answer 4. All of the following are known mechanisms of avoiding the effects
1. Describe three types of symbiotic relationships. of antibodies except
2. Describe two situations that can lead to changes in the composition a) antigenic variation.
of the normal microbiota. b) mimicking “self.”
3. How are acute, chronic, and latent infections different? c) synthesis of an Fc receptor.
4. Why are Koch’s postulates not sufficient to establish the cause of d) synthesis of IgG protease.
all infectious diseases? e) remaining intracellular.
5. Describe the four general mechanisms by which microorganisms 5. Which of the following statements about diphtheria toxin is false? It
cause disease. a) is an example of an endotoxin.
6. Describe two mechanisms that bacteria use to invade via mucous b) is produced by a species of Corynebacterium.
membranes. c) inhibits protein synthesis.
7. Explain how a capsule can allow an organism to be serum resistant d) can cause local damage to the throat.
and avoid phagocytosis. e) can cause systemic damage (that is, to organs such as the heart).
8. Give an example of a neurotoxin, an enterotoxin, and a cytotoxin. 6. Which of the following statements about botulism is true?
9. Describe two mechanisms a virus might use to prevent the induc- a) It is caused by Bacillus botulinum, an obligate aerobe.
tion of apoptosis in an infected cell. b) The toxin is heat-resistant, withstanding temperatures of 100°C.
10. How do Schistosoma species avoid antibodies? c) The organism that causes botulism can cause disease without
avoiding the immune response.
Multiple Choice d) Vaccinations are routinely given to prevent botulism.
1. Opportunistic pathogens are least likely to affect which of the fol- e) Symptoms of botulism include uncontrolled contraction of
lowing groups? muscles.
a) AIDS patients 7. Superantigens
b) Cancer patients a) are exceptionally large antigen molecules.
c) College students b) cause a very large antibody response.
d) Drug addicts c) elicit a response from a large number of T cells.
e) Transplant recipients d) attach non-specifically to B-cell receptors.
2. Capsules and M protein are thought to interfere with which of the e) assist in a protective immune response.
following? 8. Which of the following statements about endotoxin is true? It
a) Opsonization by complement proteins a) is an example of an A-B toxin.
b) Opsonization by antibodies b) is a component Gram-positive bacteria.
c) Recognition by T cells c) can be converted to a toxoid.
d) Recognition by B cells d) is heat-stable.
e) Phagosome-lysosome fusion e) causes T cells to release cytokines.
3. The C5a peptidase enzyme of Streptococcus pyogenes breaks 9. The tissue damage caused by Neisseria gonorrhoeae is primarily
down C5a, resulting in due to
a) lysis of the Streptococcus cells. a) cross-reactive antibodies.
b) lack of opsonization of Streptococcus cells. b) exotoxins.
c) killing of phagocytes. c) hydrolytic enzymes.
d) decreased accumulation of phagocytes. d) the inflammatory response.
e) inhibition of membrane attack complexes. e) all of the these.
400 Chapter 16 Host-Microbe Interactions

10. Which of the following statements about viruses is false? They 2. A microbiologist put forth a grant proposal to study the molecules
may bacteria use to communicate. Her principal rationale was that the
a) colonize the skin. damaging effects of many pathogenic microorganisms could be
b) enter host cells by endocytosis. prevented by inactivating the molecules these bacteria use to com-
c) enter host cells by fusion of the viral envelope with the cell municate. Is this a reasonable proposal? Why or why not?
membrane.
Critical Thinking +
d) induce apoptosis in infected host cells.
e) suppress expression of MHC class I molecules on host cells. 1. A student argued that no distinction should be made between
commensalism and parasitism. Even in commensalism, the micro-
Applications organisms are gaining some benefit (such as nutrients) from the
host, and this represents a loss to the host. In this sense, the host is
1. A group of smokers suffering from Staphylococcus aureus infec-
being damaged. Does the student have a valid argument? Why or
tions are suing the cigarette companies. They claim that the disease
why not?
was aggravated by smoking. The group is citing studies indicat-
ing that phagocytes are inhibited in their action by compounds in 2. A microbiologist argued that there is no such thing as “normal”
cigarette smoke. A statement prepared by their lawyers states that microbiota in the human body, since the population is dynamic
the S. aureus would not have caused such a severe disease if the and is constantly changing, depending on diet and external envi-
phagocytes were functioning properly. During the proceedings, a ronment. What would be an argument against this microbiologist’s
microbiologist was called in as a professional witness for the court. view?
What were her conclusions about the validity of the claim?
17 Immunologic Disorders
KEY TERMS
Allergy A damaging immune
response to a normally harmless
environmental antigen, generally
Hypersensitivity Exaggerated
immune response that damages
tissue; typically categorized as one
mediated by IgE. of four types.
Allergen A substance that can cause Immune Complex Combination of
an allergic reaction. antibody and soluble antigen capable
of triggering the classical pathway of
Allograft Transplanted tissue or
complement system activation.
organ from a non-identical donor of
the same species. Immunodeficiency A condition in
which components of the immune
Autoimmune Disease Damage
system provide an inadequate
caused by reactions of the immune
response; renders an individual
system against an autoantigen (“self”
susceptible to infection.
antigen).
Systemic Anaphylaxis Immediate
Delayed-Type Hypersensitivity
hypersensitivity reaction caused by
Exaggerated immune response of
IgE bound to circulating basophils.
antigen-specific T cells.
Tolerance Decreased reactivity
Desensitization Allergy treatment
of the immune system to a specific
that induces IgG production by
antigen.
gradual exposure to small amounts
of allergen; IgG competes with IgE.
Asthmatic individual using an inhaler.

A Glimpse of History
Pasteur is widely quoted as saying, “Chance favors the prepared mind.”

T
he immune system does a superb job protecting the body
This was certainly the case when physiologist Charles Richet discovered
hypersensitivities, commonly called allergies. He and his colleague, Paul from infection, but, as Richet showed, these same protec-
Portier, were cruising in the South Seas on Prince Albert of Monaco’s tive mechanisms can be harmful if uncontrolled. Just as
yacht when they hypothesized that the Portuguese man-of-war jellyfish a building’s sprinkler system that protects against fire can also
must produce a toxin that causes the swollen painful reactions to its cause significant water damage, particularly noticeable if set
stings. Prince Albert encouraged them to study the reactions, so they off by accident, immune responses can also be destructive. An
made an extract of the jellyfish tentacles and showed that it was indeed immune response that injures tissue is a hypersensitivity and can
toxic to rabbits and ducks. be categorized into one of four groups according to the mecha-
Upon returning to France, Richet and Portier continued their work nisms and timing of the response (table 17.1).
by studying the effects of the toxin from a readily available sea anemone. Various terms are helpful to describe hypersensitivities and
When they tested it on dogs, some survived the first exposure to the
other immune disorders. An allergy, or allergic reaction, is a
potent toxin, but when given a small second dose at least 3 weeks later,
hypersensitivity reaction to an allergen, a normally harmless
the dogs died within minutes. Richet and Portier had worked with toxins
and antisera earlier so they had prepared minds. They recognized that the environmental substance. Although these terms can refer to any
dogs’ reactions were probably caused by an immune response. Unlike type of hypersensitivity, they are most often used to describe
protective immune responses they had worked with before, however, those that involve an IgE response. An autoimmune disease is
this one was destructive. They called the response anaphylaxis, indicat- an inappropriate immune response that targets body tissues. An
ing it was the opposite of phylaxis, the Greek term for protection. Richet immunodeficiency is a disorder that occurs when the immune
received a Nobel Prize in 1913 for his work on anaphylaxis. system is too weak to prevent infection.

401
402 Chapter 17 Immunologic Disorders

TABLE 17.1 Some Characteristics of the Major Types of Hypersensitivities

Type I Type III Type IV
Hypersensitivity Type II Hypersensitivity Hypersensitivity
Immediate Hypersensitivity Immune Delayed-Type
Characteristic IgE-Mediated Cytotoxic Complex-Mediated Cell-Mediated
Effector B cells B cells B cells T cells
Type of antigen Soluble Cell-bound Soluble Soluble or cell-bound
Type of antibody IgE IgG, IgM IgG, IgM None
Other immune cells involved Basophils, mast cells NK cells Phagocytes Dendritic cells
Mediators Histamine, Complement, ADCC Complement, neutrophil Cytokines
leukotrienes, proteases, cytokines
prostaglandins
Time of reaction after Immediate, up to Hours to days Hours to days Peaks at 48 to 72 hours
challenge with antigen 30 minutes
Skin reaction Wheal and flare None Arthus reaction Induration, inflammation
Examples Hives, hay Transfusion reaction, Serum sickness, farmer’s Tuberculin reaction,
fever, asthma, hemolytic disease of lung, malarial kidney contact dermatitis, tissue
anaphylactic shock the newborn damage, disseminated transplant rejection
intravascular coagulation

17.1 ■ Type I Hypersensitivities: Allergic reactions occur only in sensitized individu-

als who have had prior exposure to that specific antigen.
Immediate IgE-Mediated Sensitization begins when contact with the allergen induces
an antibody response. Recall that all B cells are initially pro-
Learning Outcomes grammed to produce IgM molecules, but they can then switch
1. Describe the immunologic reactions involved in type I to produce other classes. B cells that reside in tissues under
hypersensitivities. the mucous membranes often switch to IgA production. The
2. Compare and contrast localized allergic reactions and systemic B cells can also switch to IgE production, which happens more
anaphylaxis. often in people who are prone to allergies. As IgE accumulates
3. Explain the treatments for allergies. in this area, which is rich in mast cells, the IgE molecules
attach by their Fc portion to receptors on the mast cells and
Type I hypersensitivities involve IgE, the class of antibody that also on circulating basophils (figure 17.1). Once attached, the
binds by its Fc portion to mast cells or basophils. When bound IgE molecules are stable for weeks with their antigen-binding
to the cells, the antibodies function as captured antigen recep- sites available to interact with allergens. A typical sequence
tors, allowing mast cells and basophils to detect invaders. If the of events occurs when pollen grains contact the mucous
attached IgE molecules bind an invader, the cell is triggered to membrane in an individual’s respiratory tract, triggering IgE
release inflammatory mediators, a response that is particularly production, which then sensitizes that individual to pollen.
effective against parasitic worms. Unfortunately, when IgE binds class switching, p. 366 basophil, p. 339
to allergens, the effect is the same but, like a sprinkler system set People with type I hypersensitivities have many IgE
off by accident, the response has no benefits, only drawbacks. A antibodies attached to mast cells throughout their bodies.
wide variety of common substances can cause allergic reactions, Unless exposed to the allergen recognized by these antibod-
including inhaled substances such as pollen, pet dander, and mold; ies, the mast cells are harmless. However, when at least two
ingested substances such as peanuts, milk, and seafood; and sub- cell-bound IgE molecules react with specific antigen, the IgE
stances injected into the bloodstream such as insect venom or cer- molecules come together in a reaction called cross-linking
tain drugs. mast cell, p. 340 inflammatory mediators, p. 348 IgE, p.363 (figure 17.1). Within seconds, the cross-linking causes the mast
One of the questions immunologists are investigating is why cell to release histamine and other inflammatory mediators in
some people are much more likely to develop allergies than oth- a process called degranulation. These chemicals cause capil-
ers. The tendency to have type I allergic reactions is inherited, but lary dilation, smooth muscle contraction, and increased mucus
the specific allergen to which a person reacts is due to environ- secretion. They are the direct cause of hives (urticaria), hay
mental exposures rather than genetic traits. Food allergies affect fever (allergic rhinitis), asthma, anaphylactic shock, and other
about one out of every five children, but these often go away by allergic manifestations. Symptoms begin within minutes of
the time the children reach school age. exposure to the allergen.
 Part III Microorganisms and Humans 403

First Exposure to Allergen Subsequent Exposure to Allergen

1 B-cell activation 4 Cross-linking of cell-bound IgE

Allergen B-cell receptor

Cross-linking
Cross-linking by allergen
causes the mast cell to
degranulate.
B cell

Helper T cell B cell binds to allergen and Allergen

is activated by helper T cell.

2 Proliferation, class-switching, and differentiation 5 Degranulation and release of mediators

Granules

Histamine and other

mediators are released.

Histamine
B cell

The B cells differentiate

into IgE-producing plasma
cells and memory cells.

6 Allergy symptoms

Memory cells

Plasma cells IgE antibodies

specific for allergen
Symptoms of allergy
include sneezing
and runny nose.
3 Sensitization

Fc portions of IgE antibodies

bind to mast cell receptors.

IgE receptor

Mast cell with

antigen-binding sites exposed

FIGURE 17.1 Type I Hypersensitivity: Immediate IgE-Mediated First exposure to an allergen induces an IgE antibody response,
leading to sensitization. With subsequent exposures to the allergen, cross-linking of IgE molecules on mast cells results in release of histamine and
other substances that can cause itching, swelling, and pain, and conditions such as asthma, hay fever, and anaphylactic shock.
? Why are antihistamines sometimes used to combat allergy symptoms?

mast cells. Responses in the lung may also involve constriction

Localized Allergic Reactions of smooth muscle that can interfere with gas exchange across the
Localized allergic reactions often occur in the skin or in the lungs. respiratory surface.
Typical responses in the skin include swelling due to fluid leaking Hives (urticaria) is an allergic skin condition characterized by
from capillaries that dilate as a result of histamine release from the formation of a wheal and flare; the wheal is an itchy swelling
404 Chapter 17 Immunologic Disorders

MicroByte
The most common airborne allergen in the United States is ragweed
pollen.

Systemic Anaphylaxis
Systemic anaphylaxis is a rare but serious form of IgE-mediated
allergy. It can occur when antigen enters the bloodstream and
spreads throughout the body. When this antigen binds to IgE on
circulating basophils, the cells release their inflammatory media-
tors systemically, causing extensive blood vessel dilation that
results in fluid loss from the blood. This causes a severe drop in
blood pressure that may lead to heart failure and insufficient blood
flow to the brain and other vital organs—a condition called ana-
phylactic shock. Suffocation can occur when the bronchial tubes
Wheal Flare constrict. Anaphylactic reactions may be fatal within minutes (see
A Glimpse of History).
FIGURE 17.2 The Wheal and Flare Skin Reaction A variety of Bee stings, peanuts, and penicillin injections probably account
antigens are injected or placed in small cuts in the skin to test for sen-
sitivity. Immediate wheal and flare reactions occur with antigens to
for most cases of systemic anaphylaxis. Penicillin molecules are
which the person is sensitive. changed in the body of a person with a penicillin allergy to a hap-
ten; this can react with body proteins, forming a penicillin-protein
? Are these results positive for all antigens tested?
conjugate that elicits the production of IgE antibodies against
penicillin. Fortunately, only a tiny fraction of people who make
the antibodies are prone to anaphylactic shock. Peanut allergies
generally resembling a mosquito bite, surrounded by redness, the are a problem because peanuts and their products (such as peanut
flare. The wheal and flare reaction is seen also in positive skin oil) are found in so many foods that it is often hard to predict
tests for allergens (figure 17.2). Hives may occur when a person where they will be encountered. Peanut allergies are so wide-
with a food allergy ingests that food. Allergens are absorbed from spread that peanuts have been barred by some airlines as a snack
the intestinal tract into the bloodstream where they are carried to during flights and are commonly forbidden from school lunches.
tissues such as skin. In the skin, the allergens bind to IgE carried Systemic anaphylaxis can usually be controlled by immediate
by mast cells. The mast cells degranulate, releasing inflamma- injection of epinephrine. hapten, p. 373
tory mediators, which causes swelling due to fluid accumulating
in skin tissues as it leaks from dilated capillaries. Respiratory
Treatments to Prevent
obstruction may occur if there is extensive tissue swelling in the
throat and larynx. Because histamine is a major mediator in this Allergic Reactions
situation, the reaction is blocked by antihistamine medications, Immunotherapy alters the immune responses and can be used to
such as diphenhydramine (Benadryl). decrease the negative impact of hypersensitivity reactions. One such
Hay fever, also known as allergic rhinitis, is marked by itch- treatment is desensitization (hyposensitization), a procedure that
ing, teary eyes, sneezing, and runny nose. This hypersensitivity causes the immune system to produce more IgG against the aller-
reaction occurs when people inhale an airborne antigen to which gen. These antibodies probably protect the patient by binding to the
they are sensitive. The mechanism is similar to that of hives and offending antigen, thus coating it and facilitating its removal
is also blocked by antihistamines or drugs that inhibit mast cell so that it cannot attach to IgE on mast cells or basophils
degranulation. (figure 17.3). Desensitization therapy involves injecting the
Asthma is also an immediate respiratory allergy. An allergen person with the allergen in extremely diluted, but gradually
reacting with IgE-sensitized mast cells causes their degranulation increasing concentrations over a period of several months. The
and release of inflammatory mediators into the lower respiratory antigen must be diluted enough to avoid an anaphylactic reac-
system. The mediators cause spasms of the bronchial tubes, which tion. As the concentration in the injections increases during the
interfere with breathing. Mediators other than histamine, mainly course of treatment, the individual becomes less and less sensitive
lipids such as leukotrienes and prostaglandins, are responsible for and may even lose the hypersensitivity entirely. Concurrently,
bronchospasm and increased mucus production. Eosinophils also antibodies may allow IgG to outcompete IgE antibodies for
contribute to an inflammatory response. Antihistamines are not antigen binding sites, neutralizing the antigen before it contacts
effective in treating asthma, but several other drugs are available sensitized mast cells. Activated regulatory T cells may also play a
to block the reaction, including bronchodilating drugs that relax role through release of cytokines that suppress the IgE response.
constricted muscles and relieve the bronchospasm. These are often IgG, p. 361 regulatory T cells, p. 356
self-delivered at the onset of an attack through an inhaler (see the Another allergy treatment is to prevent IgE antibodies from
chapter-opening photo). Cortisone-like steroids are often used to binding to mast cells and basophils. A medication called omali-
decrease the inflammatory reaction over a longer period of time. zumab is a genetically engineered form of an IgG molecule that
An anti-IgE therapy (described later) may also be effective. binds specifically to the Fc portion of IgE molecules, thereby
 Part III Microorganisms and Humans 405

17.2 ■ Type II Hypersensitivities:

Cytotoxic
Repeated injections
of very small amounts
Learning Outcomes
of antigen
4. Explain the mechanisms of lysis of red blood cells in transfusion
reactions.
5. Explain the cause of hemolytic disease of the newborn and how it
can be prevented.

In type II hypersensitivity reactions, antibodies react with

molecules on the surface of a cell and trigger its destruction
(a)
by the complement system or by antibody-dependent cellular
cytotoxicity (ADCC). Recall that antibodies bound to an anti-
Mast cell
gen trigger the classical pathway of complement activation,
IgE antibody and one outcome of activation is cell lysis due to membrane-
attack complexes (MACs) assembling in cell membranes. In
ADCC, the Fc regions of antibodies bound to a cell mark the
cell for destruction. Natural killer (NK) cells bind to the Fc
regions and then deliver chemicals that destroy the marked
cell. Because the responses destroy cells, type II reactions
are called cytotoxic hypersensitivities. Examples include
Antigen Granule transfusion reactions and hemolytic disease of the newborn.
containing Some autoimmune diseases also involve type II reactions.
IgG antibody mediators
complement system, p.  344 antibody-dependent cellular cytotoxicity
(ADCC), p. 361
(b)

FIGURE 17.3 Immunotherapy for IgE Allergies (a) Repeated Transfusion Reactions
injections of very small amounts of antigen are given over several
months. (b) This regimen leads to the formation of specific IgG anti- Erythrocytes (red blood cells) have various antigenic determinants
bodies. The IgG reacts with antigen before it can bind to IgE, and on their surface that can differ from one person to another. A
therefore it blocks the IgE reaction. major blood group system is reflected in ABO blood types. People
? Why are only very small amounts of antigen injected? have blood type A, B, AB, or O, depending on which specific car-
bohydrate antigens are present on their red blood cells; they have
antibodies to the antigens not present on their own red blood cells.
blocking the site that would otherwise attach to mast cells and baso- For example, people who have type A antigens are blood type A;
phils. Injections of omalizumab are effective in treating most asthma they have anti-B antibodies. Those who have type B antigens are
cases, reducing the need for more toxic medications. However, the blood type B and have anti-A antibodies. People with blood type
medication is expensive and can cause anaphylactic shock on rare O lack A and B antigens, but have antibodies to both. People with
occasions, so it is generally used only for the most serious asthma blood type AB have both A and B antigens, and have antibodies
cases. It is an example of a rhuMab (recombinant humanized to neither.
Monoclonal antibody) used in treating various other conditions. Anti-A and anti-B antibodies are called natural antibodies
because they occur without obvious pre-exposure. They probably
MicroAssessment 17.1 arise due to multiple exposures to substances similar to blood
Hypersensitivity reactions are immunological reactions that cause group antigens that occur in environmental materials such as
tissue damage. Type I hypersensitivity reactions mediated by cell- bacteria, dust, and food. Anti-A and anti-B antibodies are not
bound IgE antibodies occur immediately after exposure to antigen. present at birth but generally appear within 6 months. They are
The reactions are caused by the release of histamine and other mostly of the class IgM, and therefore cannot cross the placenta.
inflammatory mediators from cell granules. Localized allergic IgM, p. 361
reactions include hives, hay fever, and asthma; systemic reactions
When an individual receives a transfusion of red blood
can lead to anaphylactic shock. Immunotherapy is used to induce
IgG rather than IgE-mediated responses. cells with different antigens from his or her own, a transfusion
reaction occurs when natural antibodies bind to those cells, result-
1. How do localized allergic and systemic anaphylactic reactions
differ?
ing in agglutination (clumping). The transfused cells are rapidly
destroyed by membrane attack complexes or NK cells. Release
2. Define allergen, and give five common examples of substances
that act as allergens.
of hemoglobin into the bloodstream and the presence of damaged
membranes can block blood vessels and initiate clotting reactions,
3. Why does leaking of plasma from blood vessels during an
allergic reaction cause a wheal? +
damaging kidneys and producing fever along with respiratory and
digestive problems. Because of the potentially life-threatening
406 Chapter 17 Immunologic Disorders

TABLE 17.2 Antigens and Antibodies in Human ABO Blood Groups

Incidence of Blood Type in United States

Blood Antigen Present on

Type Erythrocyte Membranes Antibody in Plasma Among Whites Among Asians Among Blacks
A A Anti-B 41% 28% 27%
B B Anti-A 10% 27% 20%
AB A and B Neither anti-A nor anti-B 4% 5% 7%
O Neither Anti-A and anti-B 45% 40% 46%

consequences of giving the wrong blood to a patient, donor and To save the fetus it is sometimes necessary to transfuse it
recipient blood types are carefully cross-matched in clinical repeatedly in utero, using Rh-negative blood. However, much
settings. The cells and serum of the donor are mixed with the cells more commonly, the disease becomes life-threatening only
and serum of the recipient. Any clumping indicates an incompat- shortly after birth. Before birth, the fetus can usually survive the
ibility between the two blood types, either in the ABO group attack by anti-Rh antibodies because toxic products of red blood
(table 17.2) or in another of the many antigens on the surface of cell destruction are eliminated by maternal enzymes. However,
the red blood cell. That blood would not be used in the transfusion. the newborn baby lacks adequate levels of these enzymes, and as
soon as 36 hours after birth can become jaundiced and seriously
ill as a result of accumulation of these toxins. Immediate treatment
Hemolytic Disease is sometimes required to correct anemia and prevent permanent
of the Newborn brain damage.
Another important antigen on red blood cells is the Rh (rhesus) Hemolytic disease of the newborn is rare today because
antigen. People with the Rh antigen are Rh-positive; those with- of a medication called RhoGAM, which contains anti-Rh anti-
out are Rh-negative. In contrast to the ABO system, Rh-negative bodies. Rh-negative women who are carrying an Rh-positive
individuals do not have natural antibodies to the Rh antigen. fetus are injected with anti-Rh antibodies during pregnancy
These antibodies develop only when the individual is exposed to and again shortly after delivery. The anti-Rh antibodies bind
Rh-positive cells. Just as donor and recipient bloods are matched to any Rh-positive erythrocytes that may have entered the
for the ABO system, they are also matched for Rh system anti- mother’s circulation from the baby. This prevents these red
gens. If Rh-positive blood is given in error, an Rh-negative blood cells from stimulating a primary immune response with
recipient will then develop antibodies to the Rh-positive red the development of memory cells. Unfortunately, injecting
blood cells. If the person is exposed to Rh-positive blood again, anti-Rh antibody is not effective if the Rh-negative mother
his or her immune system will destroy the transfused cells. A is already sensitized and has formed memory cells. primary

more common event that exposes an Rh-negative person to immune response, p. 365

Rh-positive cells occurs during pregnancy.

An Rh-negative woman is likely to become sensitized to
MicroByte
the Rh antigen during childbirth if she carries an Rh-positive
Phototherapy converts toxic erythrocyte breakdown products to a
baby. Entry of Rh-positive cells into the mother’s blood- form that is more readily removed by the kidneys and excreted.
stream will cause her to develop antibodies to the Rh antigen.
Sensitization usually occurs as the mother’s blood is exposed
to the baby’s blood when the placenta ruptures during child- MicroAssessment 17.2
birth, but it can also occur during pregnancy, or after induced
or spontaneous abortion. Antibodies produced by the woman Type II cytotoxic hypersensitivity reactions are mediated by
antibodies, either by lysis of cells via the complement system or by
will not affect her red blood cells because her cells lack the
antibody-dependent cellular cytotoxicity. Blood transfusion reactions
antigen. They will not affect her first baby because IgM, which and hemolytic disease of the newborn are examples.
is produced upon first exposure, cannot cross the placenta. If
4. Describe the mechanism of cell damage in a blood transfusion
the woman carries a second Rh-positive baby, however, her where ABO antigens are mismatched.
anti-Rh IgG antibodies can cross the placenta and damage her
5. Why do Rh-negative but not Rh-positive mothers sometimes have
developing fetus, causing hemolytic disease of the newborn babies with hemolytic disease of the newborn?
(figure 17.4). This is also called erythroblastosis fetalis since
6. Why is it surprising that people who lack the A antigen on their
the affected fetus responds by producing immature red blood red blood cells have antibodies to that antigen? +
cells called erythroblasts.
 Part III Microorganisms and Humans 407

With RhoGAM

Second
Rh+ fetus

Rh– woman

Rh antigen

RhoGAM binds to fetal red blood cells leading Second Rh+ fetus. Fetal red blood
to their removal before they can stimulate the cells are not destroyed; mother
mother's immune system to make antibodies. will receive RhoGAM again.
Rh+ fetus

Without RhoGAM

First Rh+ fetus. Baby’s red blood

cells with Rh+ antigen enter
mother’s circulation during birth.

Second
Rh+ fetus

Anti-Rh IgG
antibody
FIGURE 17.4 Hemolytic Disease of the
Newborn RhoGAM contains antibodies to Rh anti-
gens and removes them from the mother’s blood
before her immune system can respond to them. Mother’s immune system reacts, Second Rh+ fetus. Mother’s IgG antibodies cross
producing antibodies to Rh. the placenta; her anti-Rh antibodies bind to the
? Would a mother need RhoGAM again with a baby’s red blood cells, resulting in their destruction.
third Rh+ fetus?

Type III hypersensitivities are caused by the formation of

17.3 ■ Type III Hypersensitivities: immune complexes consisting of IgG or IgM antibodies bound to
Immune Complex-Mediated soluble antigen. Generally, these complexes are rapidly removed
from the bloodstream, particularly if they are large. This occurs
Learning Outcomes because the Fc receptors on phagocytes bind the Fc regions of
6. Describe the role of immune complexes in type III hypersensitivity the antibodies, allowing the phagocytes to engulf and destroy the
reactions. complexes. If there is slightly more antigen than antibody, how-
7. Compare and contrast the Arthus reaction with serum sickness. ever, smaller complexes form. These are not quickly destroyed
but remain in circulation or at their sites of formation in tissue.
408 Chapter 17 Immunologic Disorders

Circulating immune complexes can lodge in blood vessel walls deposited in the kidneys, they cause glomerulonephritis. Immune
and in skin, joints, the kidneys, and other tissues where the cap- complexes can also trigger a devastating condition, disseminated
illaries are small and densely packed. They have considerable intravascular coagulation, in which clots form in small blood
biological activity due to their ability to trigger a blood-clotting vessels, leading to failure of vitalorgans. glomerulonephritis,
cascade and activate the complement system (figure 17.5). When pp. 395, 490 disseminated intravascular coagulation, p. 674
the complement system is activated, some of the components The Arthus reaction is a localized immune complex reac-
recruit phagocytes, which then release pro-inflammatory cyto- tion. If antigen is injected into a previously immunized person
kines. The phagocytes may also release enzymes and toxic mol- who already has high levels of circulating specific antibody,
ecules that damage surrounding tissue. Fc receptors, p. 390 immune complexes can form at the site of injection where anti-
Immune complex disease may arise during a variety of bacte- gen levels are high. The result is a reaction characterized by
rial, viral, and protozoan infections, as well as from inhaled dust severe pain, swelling, and redness. This can happen, for example,
or bacteria, and injected medications such as penicillin. They when tetanus-diphtheria booster vaccine is given to a person
are responsible for the rashes, joint pains, and other symptoms too frequently. The immune complexes form outside the blood
seen in a number of diseases, such as farmer’s lung, lupus, bac- vessels, in the skin tissues, and activate complement, producing
terial endocarditis, early rubella infection, and malaria. When complement components that attract neutrophils. The release of

1 Antigen in excess

4 Circulating immune complexes are trapped in the vessel walls.

Antigen

Antibody

Endothelial cells Clump of immune

of blood vessel complexes

2 Formation of immune complexes

5 Neutrophils are attracted and release enzymes that cause

tissue damage.

Immune
complex

Neutrophils release
enzymes.
3 Activated complement causes basophils to release inflammatory
mediators that increase vascular permeability.

Complement
activated
Basophil
Mediators
FIGURE 17.5 Type III Hypersensitivity: Immune
Complement Complex-Mediated When antigen is in slight excess (1), immune
in the blood complexes form and activate complement (2), resulting in increased
vascular permeability (3). Immune complexes are trapped in the blood
Spaces created
by dilation of vessels (4) and complement attracts neutrophils that release enzymes,
blood vessel damaging the tissue (5).

? Why do immune complexes form only when an antigen is soluble?

 Part III Microorganisms and Humans 409

neutrophil enzymes contributes to a local inflammatory response mycobacterial protein antigens are injected into the skin, people
that peaks in 6 to 12 hours. who are infected with the organism will exhibit a delayed-type
Serum sickness is a systemic immune complex disease hypersensitivity reaction. The site of injection reddens and gradu-
caused by passive immunization (antibodies provided as a ally becomes indurated (thickened) within 6 to 24 hours, reach-
source of protection). When an antibody-containing serum ing a peak in 2 to 3 days (figure  17.6). No wheals (as seen in
from a horse or other animal is injected into humans to pre- IgE-mediated reactions) form. The reaction results mainly from
vent or treat a disease, the recipient may mount an immune effector helper T cells that recognize the antigens and then release
response to antigens in the foreign serum. After 7 to 10 days, pro-inflammatory cytokines. The inflammatory response that
enough immune complexes form to cause signs such as fever, characterizes a positive skin test also occurs in response to the
inflammation of blood vessels, arthritis, and kidney damage. infecting organisms, often resulting in the formation of tubercles
People with serum sickness usually recover as the antigens of in the lung.
the animal serum are cleared from the body. Because of the
potential to cause serum sickness, horse serum is no longer
used for passive immunization against diphtheria and tetanus; Delayed-Type Hypersensitivity
instead, hyperimmune human globulin is used. The serum in Infectious Diseases
sickness form of hypersensitivity is rare now, but can occur Recall from chapter 15 that cell-mediated immunity plays a cen-
following treatment of heart attack patients with the bacterial tral role in combating intracellular microbial infections. Effector
enzyme streptokinase to dissolve clots, or after injection of cytotoxic T cells are particularly important because they destroy
horse-derived anti-venom to treat snakebites. passive immunity, the infected host cells. Although this prevents the infection from
p. 420 hyperimmune globulin, p. 420 spreading, it also damages the tissues. With chronic infections,
delayed-type hypersensitivity causes extensive host cell destruc-
MicroByte tion and progressive loss of tissue function, as seen in the dam-
Like farmer’s lung that develops from inhaling moldy hay, librarian’s
aged sensory nerves of leprosy. The immune response is indeed
lung can develop in response to inhaled mold and dust from books.
a two-edged sword, with a delicate balance between protective
immunity and harmful hypersensitivity. cell-mediated immunity,
MicroAssessment 17.3 p. 355 leprosy, p. 650

Type III immune complex–mediated hypersensitivities arise when

small complexes of antigen-antibody persist in the tissues and activate
the complement system, triggering an inflammatory response.
Contact Hypersensitivities
7. How are immune complexes normally cleared from the body? Contact hypersensitivity, also called contact dermatitis or contact
allergy, is due to effector T cells responding to small molecules
8. How can immune complexes cause tissue damage?
that penetrate intact skin. The mechanisms of tissue damage are
9. Why are the kidneys particularly vulnerable to immune complex
similar to those of other types of delayed-type hypersensitivities,
damage? +

17.4 ■ Type IV Hypersensitivities:

Delayed-Type Cell-Mediated
Learning Outcomes
8. Describe the key immunologic reactions involved in type IV
hypersensitivities.
9. Outline the role of type IV hypersensitivity reactions in contact
dermatitis or contact allergies.

Delayed-type hypersensitivity reactions (type IV hypersensi-

tivities) are due to antigen-specific T-cell responses and can occur
almost anywhere in the body. Whereas immediate hypersensi-
tivities involve antibodies and occur within minutes, these cell-
mediated reactions peak 2 to 3 days following antigen exposure.
cell-mediated immunity, p. 355
FIGURE 17.6 Positive Skin Test for Delayed-Type
Hypersensitivity Reaction Injection of tuberculin protein into the
skin of a person sensitized to Mycobacterium tuberculosis causes a
Tuberculin Skin Test firm red raised area to form by 48 to 72 hours. A reaction greater
The tuberculin skin test, which can be used to detect latent than 10 mm in diameter is considered positive.
Mycobacterium tuberculosis infections, relies on a delayed- ? How many days after a tuberculin skin test should results be
type hypersensitivity reaction. When a very small quantity of evaluated?
410 Chapter 17 Immunologic Disorders

but damage to skin cells causes an irritating rash and sometimes

blisters (figure 17.7).
Familiar substances like the nickel of metal jewelry, the
chromium salts in certain leather products, or components of
some cosmetics are common culprits in contact dermatitis. An
oily product of poison ivy and poison oak is another example
of a substance that causes the reaction (figure  17.8). All of
these materials shed small chemicals that act as haptens and
combine chemically with proteins in the skin. Dendritic cells
take up and process these hapten:protein complexes and pres-
ent the resulting hapten:peptides to T cells in nearby lymph
nodes. T cells that become activated form memory cells,
sensitizing the individual to the substance. Once sensitized, a
second exposure causes a damaging cell-mediated response.
hapten, p. 373
FIGURE 17.7 Severe Contact Hypersensitivity Note the Latex products are a frequent cause of hypersensitiv-
redness and blisters.
ity reactions. Latex, a product of the rubber tree, con-
? What type of immune cells cause this reaction? tains a plant protein that readily induces sensitization.

FIGURE 17.8 Poison Oak Dermatitis

This skin reaction results from delayed-type
hypersensitivity. A hapten from poison oak
? Why does a sensitized person combines with skin proteins.
not experience a skin reaction
immediately after exposure to First exposure Subsequent exposures
poison oak? activate memory cells

1 Dendritic cells present the hapten:peptide complex to T cells. 3a Helper T cells activate 3b Cytotoxic T cells destroy
macrophages presenting skin cells presenting
Hapten:peptide hapten:peptide complex. hapten:peptide complex.

Dendritic cell Naive helper Naive cytotoxic

T cell T cell Macrophage Skin cell

2 T-cell activation, proliferation, and differentiation increases the Cytotoxic T cell

population of T cells that recognize the hapten:peptide complex. Cytokines Helper T cell
Naive helper Naive cytotoxic
T cell T cell
4 Activated macrophages 5 A rash appears after 24 hours,
release inflammatory mediators. reaching a peak at 48–72 hours
after exposure.

Effector
T cells

Macrophage

Memory
cells Inflammatory
mediators
 Part III Microorganisms and Humans 411

Many products contain latex, such as fabrics, elastics, toys, Transplant rejection is predominantly a cell-mediated immu-
and contraceptive condoms, but latex gloves probably account nological reaction, involving a complex set of events centered
for most latex sensitization. Gloves are used extensively by around the actions of effector cytotoxic T cells and natural killer
healthcare and laboratory workers, food preparers, and many (NK) cells. To avoid rejection, antigen incompatibilities are
others. Typically, a person will notice redness, itching, and a minimized by using MHC tissue typing and ABO blood typing
rash on the hands after wearing gloves. To prevent the reac- to match donor and recipient tissues. Even in well-matched tis-
tion, latex gloves should be replaced by vinyl or other syn- sue transplants, however, recipients must use immunosuppres-
thetic gloves. Topical cortisone-like medications that inhibit sive drugs indefinitely to prevent graft rejection. These drugs are
T-cell responses are effective treatments. needed to prevent immune reaction to the many minor antigens
The causative substance in contact hypersensitivity is com- present on allograft cells. cytotoxic T cells, p. 357 NK cells, p. 374
monly detected by patch tests, in which suspect substances are Immunosuppressive drugs minimize the rejections, but they
applied to the skin under an adhesive bandage. Positive reactions make the recipient more susceptible to infections and cancer.
consisting of redness, itching, and blisters reach their peak in Cyclosporin A (produced by a fungus) and tacrolimus (produced
about 3 days. from a species of Streptomyces) are both effective immunosuppres-
sants. These drugs interfere with cellular signaling, thereby inhib-
MicroByte iting clonal selection and expansion of activated T lymphocytes.
Approximately half of the people with latex allergies are allergic to They suppress only T-cell proliferation, so the patient still has
bananas as well.
some level of immunity mediated by other types of immune cells.
clonal selection and expansion, p. 363

MicroAssessment 17.4 Many thousands of bone marrow transplants have saved the
lives of individuals with cancers such as multiple myeloma and
Type IV delayed-type hypersensitivity depends on the action of
sensitized T cells. The reaction peaks 2 to 3 days after exposure to
leukemia, and other serious conditions. Generally, the patient’s
antigen. Examples are contact dermatitis, damage in a variety of bone marrow and immune system are intentionally destroyed by
infectious diseases, rejection of tissue grafts, and some autoimmune intense radiation and chemotherapy before transplantation of the
diseases. new bone marrow. Donor bone marrow is given to the recipient by
10. Explain the events that occur in the skin during a positive infusing it through the large subclavian vein under the left collar
delayed-type hypersensitivity skin test. bone. The marrow hematopoietic stem cells circulate until they
11. Describe a patch test for contact hypersensitivity. reach the recipient’s bone marrow, where they leave the blood-
12. In the tuberculin skin test, why does a positive result indicate stream and begin producing hematopoietic cells. Close monitoring
prior exposure to M. tuberculosis? + is required for months after a transplant because the recipient is
prone to infection and bleeding while the grafted marrow restores
the supply of erythrocytes, leukocytes, and platelets. Recovery
17.5 ■ Rejection of Transplanted may be complicated by a graft-versus-host reaction in which the
graft-derived immune system recognizes the patient’s tissues as
Tissues an invader and attacks them. Therefore, many of these patients
also require immunosuppressive treatment. hematopoietic stem
Learning Outcomes
cells, p. 338
10. Explain the process of rejection of transplanted tissues.
11. Describe the mode of action of medications used to prevent MicroByte
rejection of transplanted tissue. Hematopoietic stem cells can sometimes be filtered from donor
blood without the use of general anesthetic or punctures into the
A special case of delayed-type cell-mediated hypersensitivity marrow.
occurs with the rejection of transplanted organs or tissues. Most
human transplants involve allografts in which the tissues of the
MicroAssessment 17.5
donor and recipient are not genetically identical. Antigenic dif-
ferences, particularly in the major histocompatibility complex Successful organ and tissue transplantation depend on matching major
(MHC) molecules, lead to rejection of the graft (see Perspective histocompatibility antigens and using immunosuppressive agents
15.1). Larger differences result in stronger immune reactions. such as cyclosporin and tacrolimus to minimize the immune response
to other antigens of the graft. Rejection of transplants is complex,
Certain types of transplants, including autografts (tissue trans- but type IV cellular immune responses are the major mechanism of
planted from elsewhere on the recipient’s body) and isografts rejection for allografts.
(grafts donated by an identical sibling), avoid these problems.
13. What differences make an allograft more likely to be rejected
Xenografts, involving tissues from pigs or non-human animals, than an autograft?
typically evoke vigorous immune responses, but may become
14. How is graft-versus-host disease difference from a recipient’s
more widely used as we gain the ability to genetically engineer rejection of donor tissue?
animals that express human MHC molecules. Perspective 17.1
15. Why is the recipient of a transplanted kidney unlikely to suffer
discusses a very special transplantation situation: the fetus as an from a graft-versus-host reaction? +
allograft. MHC molecules, p. 369
412 Chapter 17 Immunologic Disorders

PERSPECTIVE 17.1
The Fetus as an Allograft
One allograft that does not face the usual threat used for typing major histocompatibility anti- or II molecules and is not subject to T-cell
of rejection is a very special one—the mam- gens can be obtained from women who have attack. It also avoids destruction by natural
malian fetus. Half of the fetal antigens are of had several children with the same father— killer cells. A few other areas in the body—the
paternal origin and are likely to differ from they have made antibodies to his MHC mol- brain, the eyes, and the testes—are also immu-
those of the mother. In spite of immunological ecules. Furthermore, small numbers of fetal nologically privileged sites. Antigens leaving
differences, the fetus develops in the uterus for cells are found in the maternal circulation these sites do not drain through lymphatic ves-
9 months and is not rejected. The mechanisms during pregnancy. The placenta, however, sels and reach lymphoid tissues where antigen-
for survival of the fetal allograft have long prevents most fetal cells from entering the presenting cells are abundant. Also, antigen
been the subject of research, but they are not mother and most maternal T cells from reach- leaves these privileged sites accompanied by
yet fully understood. ing the fetus. immunosuppressive cytokines that direct the
Do paternal antigens from the fetus reach The fetus is protected in the uterus, an immune response toward tolerance rather than
the mother’s immune system where they might immunologically privileged site. The outer layer destruction. It is well recognized that mater-
cause a response? Clearly, they do. Mothers of the placenta, the trophoblast, acts as a buf- nal immune responses are suppressed to some
make antibodies to paternal antigens, such as fer zone between the fetus and the mother. extent during pregnancy, though the reasons for
the Rh antigen on red blood cells. Antibodies The trophoblast does not express MHC class I this are not clear.

17.6 ■ Autoimmune Disease inflammation and damage even in the absence of the pathogen.
Autoimmune responses may also occur after injury in which
Learning Outcomes self antigens are released from privileged (isolated) or damaged
12. Define autoimmunity, and explain some possible causes.
organs and stimulate production of antibodies to them (autoanti-
bodies). regulatory T cells, p. 356
13. Give five examples of autoimmune diseases and the mechanism of
tissue injury in each.
The Spectrum of Autoimmune Diseases
The immune system generally eliminates or silences develop-
Autoimmune diseases can be organ-specific or systemic, depend-
ing lymphocytes that would respond to and attack normal body
ing on whether the autoantigens are localized to a specific organ
tissues, or autoantigens (“self” antigens). Autoimmune disease
or are found throughout the body (table 17.3). In both situations,
results when the system fails to do this, resulting in attacks against
however, the damage may be caused by antibodies, cell-mediated
autoantigens as if they were invaders.
immune mechanisms, or both. Examples include the following:
The causes of autoimmune disease are not clear and prob-
ably involve many things. A deficiency in the action or control of ■ Type 1 diabetes mellitus. Also known as insulin-dependent
regulatory T cells (Treg) that normally inhibit responses to “self” or juvenile diabetes, this is an organ-specific autoimmune
may be at fault. A genetic component, perhaps related to major disease caused when cytotoxic T cells destroy a group of
histocompatibility molecules, may be a predisposing factor, but pancreatic cells called β cells. These cells produce insulin, a
disease occurrence is also influenced by environmental factors, protein that allows various cell types in the body to take up
including infection. In some cases, a pathogen’s ability to evade glucose from the bloodstream. The lack of insulin results in
the immune system by mimicking “self” likely plays a role. As a increased blood levels of glucose, causing water to be drawn
result of this molecular mimicry, the immune response mounted from the cells. In turn, this leads to symptoms of increased
against the pathogen attacks healthy tissues as well, causing thirst and urination. Because the cells do not take in glucose

TABLE 17.3 Characteristics of Some Autoimmune Diseases

Disease Organ Specificity Major Mechanism of Tissue Damage

Type 1 diabetes mellitus Pancreas T-cell destruction of pancreatic β cells

Graves’ disease Thyroid Autoantibodies bind thyroid-stimulating hormone
receptor, causing overstimulation of thyroid
Systemic lupus erythematosus Systemic Autoantibodies to DNA and other nuclear components
form immune complexes in small blood vessels
Myasthenia gravis Muscle Autoantibodies bind to acetylcholine receptor on muscle,
preventing muscle contraction
Rheumatoid arthritis Systemic, especially joints Lymphocyte destruction of joint tissues; immune complexes
of IgG and anti-IgG
 Part III Microorganisms and Humans 413

that would otherwise be used as their energy source, symp-

toms also include extreme hunger, fatigue, and weight loss.
Persons with diabetes are vulnerable to high blood pressure,
stroke, blindness, limb amputation, and kidney disease.
Insulin injections are used to treat the disease.
■ Graves’ disease. This is an organ-specific autoimmune disease
that affects the thyroid gland. In most cases, antibodies are
directed at receptors on the thyroid gland for thyroid-stimulating
hormone (TSH). Attachment of the antibody to the receptor
activates the receptor inappropriately, leading to increased
thyroid hormone production and enlargement of the gland.
An enlarged thyroid is sometimes evident as a goiter (figure
17.9). Metabolic rate increases in affected individuals, result-
ing in symptoms that include weight loss, fatigue, irritability,
heat intolerance, rapid heartbeat, and bulging eyes. FIGURE 17.10 Rheumatoid Arthritis Autoimmune reactions
■ Systemic lupus erythematosus (SLE). As the name implies, cause chronic inflammation and destruction of the joints.
this is a systemic disease. Antibodies made against molecules ? What type of hypersensitivity has damaged the joints in this hand?
found in the nuclei of cells provoke an autoimmune response
throughout the body. Symptoms vary considerably, but
■ Myasthenia gravis. The name means “grave muscle weak-
typically include joint pain, swelling in the joints, and rashes.
Severe cases involve damage to the kidneys or other organs. ness” and reflects the fact that this systemic disease is caused
Curiously, the disease affects mainly women. by a disruption in nerve transmission to muscles. Antibodies
bind to acetylcholine receptors at the neuromuscular junc-
tion, thereby blocking transmission of impulses that normally
cause muscle contraction. Moreover, binding of the antibod-
ies may activate the complement system, which then damages
the acetylcholine receptors. Drugs that inhibit the enzyme
cholinesterase (the enzyme that degrades acetylcholine) allow
acetylcholine to accumulate at the neuromuscular junction
and may lessen the effects of the disease. Immunosuppressive
medications and removal of the thymus gland are helpful in
many cases. The role of the thymus in this disease is not well
understood.
■ Rheumatoid arthritis. This is a systemic disease in which
antibodies and cellular mechanisms target collagen in connec-
tive tissues, most often within joints (figure 17.10). T cells
infiltrate the joints and, when stimulated by specific antigens,
they release cytokines that cause inflammation. Antibodies to
collagen form immune complexes characteristic of type III
hypersensitivities that further damage the joint tissue. This
crippling inflammatory condition is one of the most common
autoimmune diseases, and unlike arthritis that arises from
wear and tear of joints, it may occur at any age. Rheumatoid
arthritis is most common in women ages 30 to 50. cytokines,
p. 341 immune complexes, p. 395

MicroByte
Babies born to mothers with myasthenia gravis experience temporary
muscle weakness, since IgG antibodies cross the placenta.

Treatment of Autoimmune Diseases

Autoimmune diseases are usually treated with anti-inflammatory
FIGURE 17.9 Goiter A goiter may result from overstimulation of medications or immunosuppressant drugs that interfere with
the thyroid gland. T-cell signaling or kill dividing cells, thereby limiting the immune
? What receptors in the thyroid gland are stimulated in Graves’ response. Replacement therapy may be necessary to restore dam-
disease? aged tissues or tissue products, such as when patients with type 1
414 Chapter 17 Immunologic Disorders

diabetes receive insulin. Because the risk of infection and cancer In contrast to hypersensitivities and autoimmune diseases that
increase when the immune system is suppressed, the options arise from an overblown or inappropriate immune response,
described next offer more promising approaches to the treatment immunodeficiencies arise when the body cannot make or sustain
of autoimmune disease. an immune response. Immunodeficiency disorders may be primary
A strategy being explored to treat autoimmune diseases is to (congenital) or secondary (acquired). Primary immunodeficiency
induce tolerance, a process that decreases the reactivity of the immune can result from a genetic defect or from environmental factors that
system to a specific antigen. One promising approach is to introduce cause developmental abnormalities. Secondary immunodeficiency
the antigen by the oral route, so that the immune system “learns” to can be acquired as the result of infection or other stresses on the
tolerate it, just as it does the many antigens ingested in food. This immune system such as malnutrition. People with either type of
antigen-specific immunotherapy causes fewer side effects than drugs immunodeficiency are subject to repeated infections. The types of
that generally suppress the immune system. Trials designed to induce these infections will often depend on which part of the immune
oral tolerance in humans have successfully relieved the symptoms of system is affected. Some of the more important immunodeficiency
several autoimmune diseases, but there is much still to learn about the diseases are listed in table 17.4.
immunological mechanisms, antigen preparations, doses, and dura-
tion of treatment. immunological tolerance, p. 355
Attempts have been made to cure type 1 diabetes by replac- Primary Immunodeficiencies
ing the tissues destroyed by immune cells. Transplantation of the Primary immunodeficiencies are generally rare. They may
pancreas or insulin-producing cells of the pancreas has been suc- affect B cells, T cells, natural killer (NK) cells, phagocytes,
cessful in many cases, but dangerous immunosuppressive agents or complement components. Many of the gene defects that
must be given to prevent rejection. Because of this, typically the cause primary immunological disorders are known and work is
only patients who are given pancreas transplants are those with underway to correct them. Deficiencies and defects can occur
advanced diabetes who also require a kidney transplant, so must take at any point in the complex steps that lead to an effective
the immunosuppressive drugs anyway. Research efforts are directed immune response. A few categories of primary immunodefi-
toward developing better methods of transplantation, such as inject- ciencies include the following:
ing pancreas cells harvested from cadavers into the vein that carries
■ Antibody deficiencies. One of the most common primary
blood to the liver. The cells establish themselves in the liver and
immunodeficiencies known is selective IgA deficiency, in
produce insulin, often eliminating or decreasing the need for injected
which very little or no IgA is produced. Studies indicate that it
insulin. However, the cells usually die in months or a few years, and
occurs as often as one per 333 to 700 people. Although people
the immunosuppressant side effects are often severe. Research on
manipulating stem cells to produce insulin-secreting β cells holds
greater promise for a cure of type 1 diabetes.
TABLE 17.4 Immunodeficiency Diseases
MicroAssessment 17.6
Part of the Immune
Autoimmune disease can result when the immune system attacks
autoantigens. Damage from autoimmune reactions may be caused by Disease System Involved
antibody action or by cell-mediated immune functions, or both. Some Primary Immunodeficiencies
autoimmune diseases are organ-specific, but others are systemic.
Typical treatment involves drugs that suppress immune responses, but Severe combined Bone marrow stem cells
these have dangerous side effects. immunodeficiency (SCID) (defect)
Selective IgA deficiency B cells making IgA (deficiency)
16. What might make an autoimmune reaction general rather than
tissue-specific? Congenital B cells (deficiency)
agammaglobulinemia
17. What are the negative side effects of taking immunosuppressive
drugs? Infantile X-linked Early B cells (deficiency)
agammaglobulinemia
18. How could viruses or bacteria potentially cause autoimmune
disease? + DiGeorge syndrome T cells (deficiency)
Chediak-Higashi disease Phagocytes (defect)
Chronic granulomatous Phagocytes (defect)
17.7 ■ Immunodeficiency disease (CDG)
Hereditary angioneurotic Complement regulator
Disorders edema (deficiency)
Secondary Immunodeficiencies
Learning Outcomes
Acquired immunodeficiency T cells (destroyed by viral
14. Contrast the two main categories of immunodeficiency disorders.
syndrome (AIDS) infection)
15. Explain how immunodeficiency can lead to multiple and unusual
Monoclonal gammopathy B cells (multiply out of control)
infections.
 Part III Microorganisms and Humans 415

with this disorder may appear healthy, many have repeated late components (C5, C6, C7, C8) have recurrent Neisseria
bacterial infections of the respiratory, gastrointestinal, and infections, because the membrane attack complexes typi-
genitourinary tracts, where secretory IgA normally protects cally destroy these bacteria. People who lack one of the
against colonization or invasion by pathogens. Another anti- important control proteins of the sequence, C1-inhibitor,
body deficiency is agammaglobulinemia, a disease in which experience uncontrolled complement activation. This causes
few or no antibodies are produced. fluid accumulation and potentially fatal tissue swelling, a
condition called hereditary angioneurotic edema. comple-
■ Lymphocyte deficiencies. A disease called severe com-
ment system, p. 344
bined immunodeficiency (SCID) results when hematopoi-
etic stem cells in the bone marrow produce neither T nor
B lymphocytes. Children with SCID generally die of infec- MicroByte
tious disease at an early age unless they are success- SCID is sometimes called “bubble boy disease” after a patient who
survived for 12 years in a sterile inflated plastic bubble.
fully treated with a bone marrow transplant to reconstitute
the bone marrow with healthy cells. A variety of gene defects
can cause SCID. One is a mutation in an enzyme necessary
to form B- and T-cell receptors required for antigen recogni- Secondary Immunodeficiencies
tion. Another is in a gene for the interleukin-2 receptor on
Secondary, or acquired, immunodeficiency diseases may result
lymphocytes, preventing the cells from receiving the signal
from malignancies, advanced age, pregnancy, certain infections
to proliferate. Other individuals with SCID lack adenosine
(especially viral infections), immunosuppressive drugs, or mal-
deaminase, an enzyme important in the proliferation of B
nutrition. Some viral infections will deplete certain cells of the
and T cells. A few children with SCID have responded well
immune system. The measles virus, for example, replicates in
temporarily to receiving their own defective T cells, with
various cells of the immune system, killing many of them and
an inserted adenosine deaminase gene linked to a retrovirus
leaving the body temporarily open to other infections. Syphilis,
vector. Unfortunately, the genetically altered cells do not live
leprosy, and malaria affect the T-cell population and also macro-
long, and the treatments must be repeated. Still, these results
phage function, causing defects in cell-mediated immunity.
are promising, and the possibility of treating other severe
One of the most serious and widespread secondary immu-
disorders with gene therapy is exciting.
nodeficiencies is AIDS (acquired immunodeficiency syndrome),
In children with DiGeorge syndrome, lymphocyte
caused by human immunodeficiency virus (HIV). This retrovirus
deficiency results when the thymus fails to develop in the
infects and destroys helper T cells, leaving the affected person
embryo. As a result, T cells do not differentiate and are absent
highly susceptible to infections. Opportunistic infections become
in circulation. Affected individuals have other developmental
more common, caused by agents that are usually unable to estab-
defects as well, such as heart and blood vessel abnormalities,
lish infection in a healthy individual. AIDS and opportunistic
and a characteristic appearance with low-set deformed ears,
infections are covered in chapter 28. retrovirus, p. 320
small mouth, and wide-set eyes. As expected from a lack of T
Cancers involving the lymphatic system often decrease
cells, affected people are susceptible to infections by eukary-
effective antibody-mediated immunity. For example, multiple
otic pathogens, such as Pneumocystis jiroveci and other
myeloma is a malignancy arising from a single plasma cell that
fungi, as well as viruses and obligate intracellular bacteria.
proliferates out of control and in most cases produces large
Pneumocystis jiroveci, p. 708
quantities of immunoglobulin. This overproduction of a single
■ Defects in phagocytic cells. Chronic granulomatous disease kind of molecule results in the body using its resources to produce
(CGD) is caused by a defect that results in failure of lym- immunoglobulin of a single specificity at the expense of others
phocytes to produce hydrogen peroxide and certain other needed to fight infection. The result is an overall immunodefi-
active products of oxygen metabolism. These phagocytes ciency. Other lymphoid disorders include macroglobulinemia
are unable to kill some organisms, especially the catalase- (overproduction of IgM) and some forms of leukemia.
positive Staphylococcus aureus. In Chediak-Higashi disease,
lysosomes in the phagocyte lack certain enzymes and cannot
MicroAssessment 17.7
destroy phagocytized bacteria. People with this condition
suffer from recurring pyogenic (pus-forming) bacterial infec- Primary immunodeficiencies may be caused by genetic or
tions. In leukocyte adhesion deficiency, white blood cells fail developmental defects in components of the immune response.
to leave the circulation to concentrate at sites of infection. Secondary immune deficiencies result from infection or environmental
influences.
catalase test, p. 243
19. Compare and contrast severe combined immunodeficiency
■ Defects in complement system components. People with disease (SCID) and chronic granulomatous disease (CGD).
deficiencies in the early components of the complement 20. How can a person with multiple myeloma be immunodeficient?
system (such as C1 and C2) may develop immune complex
21. How could you determine whether an immunodeficiency disease
diseases, because these components normally help clear affects T lymphocytes, B lymphocytes, or macrophages? +
immune complexes from the circulation. People who lack
416 Chapter 17 Immunologic Disorders

FUTURE CHALLENGES 17.1

New Approaches to Correcting Immunologic Disorders

In recent years, many of the genes responsi- be met, at least in part, by the development of A promising and challenging area is the
ble for immunodeficiency diseases have been gene transfer technology and by better under- development of human stem cell research.
identified. It has been possible to correct some standing of the mechanisms of cell-mediated Stem cells have an almost unlimited capacity
of these gene defects in cells in the laboratory immunological mechanisms. to divide, and some of them can differentiate
and, rarely, in patients. In the near future, One interesting line of research stems into most of the tissues in the body. They could
research will be directed toward developing from the observation that parasitic worm infes- be used to generate cells for transplantation
the existing technology for gene transfer to tations appear to protect people from allergies and to replace defective or injured tissues such
make it more effective in correcting these and autoimmune diseases. Experiments with as nerve tissue. They might also be used to test
gene defects in human patients. It is important mice support this idea and also show that the effects of drugs on human cells, without
also to continue the search for other defec- feeding mice parasitic worms effectively treats the danger of testing on human beings. A
tive genes; with increasing knowledge of the experimental autoimmune disease. The effect major stumbling block is the fact that the stem
human genome it is likely that more will be appears to be due at least in part to down- cells come from fetal material, either from
found soon. A continuing challenge is finding regulation of the immune response. Research early stage embryos obtained from fertility
ways to overcome graft rejection to make bone into understanding the regulation of immune treatments or from aborted fetuses. The legal
marrow and other transplants more acceptable. responses will help in controlling autoimmune and ethical guidelines for the use of these cells
The challenge of treating cancer and of pre- and immunodeficiency diseases, and it will are matters of considerable debate.
venting rejection of essential transplants may permit development of improved vaccines.

Summary
17.1 ■ Type I Hypersensitivities: 17.3 ■ Type III Hypersensitivities:
Immediate IgE-Mediated (table 17.1) Immune Complex-Mediated (table 17.1)
IgE attached to mast cells or basophils reacts with specific antigen, Type III hypersensitivity reactions are mediated by small
resulting in the release of powerful mediators of the allergic reaction antigen-antibody complexes that activate complement and other
(figure 17.1). inflammatory systems, attract neutrophils, and contribute to inflam-
mation. The immune complexes are often deposited in small blood
Localized Allergic Reactions
vessels in organs, where they cause inflammatory disease—for
Localized allergic (type I) reactions include hives (urticaria), hay fever example, glomerulonephritis in the kidney or arthritis in the joints
(allergic rhinitis), and asthma (figure 17.2). (figure 17.5).

Systemic Anaphylaxis
17.4 ■ Type IV Hypersensitivities:
Systemic anaphylaxis is a rare but serious reaction that can lead to
shock and death. Delayed-Type Cell-Mediated (table 17.1)
Delayed-type hypersensitivity reactions depend on the actions of sen-
Treatments to Prevent Allergic Reactions sitized T lymphocytes.
Desensitization, or immunotherapy, is often effective in decreasing
Tuberculin Skin Test (figure 17.6)
the type I hypersensitivity state (figure 17.3). A new treatment, using an
engineered anti-IgE, promises to be effective in treating asthma. A positive reaction to tuberculin protein introduced under the skin peaks
2 to 3 days after exposure to antigen.
17.2 ■ Type II Hypersensitivities: Cytotoxic (table 17.1) Delayed-Type Hypersensitivity in Infectious Diseases
Type II hypersensitivity reactions, or cytotoxic reactions, are caused
Delayed-type hypersensitivity is important in responses to many
by antibodies that can destroy normal cells by complement lysis or by
chronic, long-lasting infectious diseases.
antibody-dependent cellular cytotoxicity (ADCC).
Contact Hypersensitivities
Transfusion Reactions (table 17.2)
Contact allergy, or contact dermatitis, occurs frequently in response to
The ABO blood group antigens have been the major cause of
substances such as poison ivy, nickel in jewelry, and chromium salts in
transfusion reactions.
leather products (figures 17.7, 17.8).
Hemolytic Disease of the Newborn (figure 17.4)
The Rhesus blood group antigens are usually responsible for this poten- 17.5 ■ Rejection of Transplanted Tissues
tially fatal disease. Injected anti-Rh antibody helps prevent Rh sensitiza- Transplantation rejection of allografts is caused largely by type IV
tion of Rh-negative mothers. cellular reactions.
 Part III Microorganisms and Humans 417

17.6 ■ Autoimmune Disease Primary Immunodeficiencies

Responses against autoantigens can lead to autoimmune diseases. B-cell immunodeficiencies result in diseases involving a lack of anti-
The Spectrum of Autoimmune Diseases
body production, such as agammaglobulinemias and selective IgA
deficiency. T-cell deficiencies result in diseases such as DiGeorge syn-
Autoimmune diseases can be organ-specific (table 17.3, figure  17.9) or
drome. Lack of both T- and B-cell functions results in combined immu-
widespread (figure  17.10). Some autoimmune diseases are caused by
nodeficiencies, which are generally severe. Defective phagocytes are
antibodies produced to body components, and others by cell-mediated
found in chronic granulomatous disease and Chediak-Higashi disease.
reactions or a combination of antibodies and immune cells.
Secondary Immunodeficiencies
Treatment of Autoimmune Diseases
Acquired immunodeficiencies can result from malnutrition, immuno-
Autoimmune diseases are usually treated with drugs that suppress the
suppressive agents, infections (such as AIDS), and malignancies such
immune and/or inflammatory responses.
as multiple myeloma.
17.7 ■ Immunodeficiency Disorders (table 17.4)
Immunodeficiencies can be primary genetic or developmental defects
in any components of the immune response, or they can be secondary
and acquired.

Review Questions
Short Answer 3. All of the following are true of immune complexes except
1. Why are antihistamines useful forr treating many IgE-mediated a) the most common complexes consist of antigen and IgE.
allergic reactions but not effective in
n treating asthma? b) an immune complex consists of soluble antigen attached to
antibody.
2. Penicillin is a very small molecule, yet it can cause any of the types
of hypersensitivity reactions, especially
ecially type I. How can this c) complement components are activated by antigen-antibody
occur? complexes.
d) immune complexes cause strong inflammatory reactions.
3. What are some major differences between an IgE-mediated skin
e) immune complexes deposit in kidneys, joints, and skin.
reaction, such as hives, and a delayed-type
d-type hypersensitivity reaction,
such as a positive tuberculin skin test?t? 4. Delayed-type hypersensitivity reactions in the skin
4. What causes insulin-dependent diabetesetes mellitus? a) are characterized by a wheal and flare reaction.
b) peak at 4 to 6 hours after exposure to antigen.
5. Compare and contrast the autoimmune une processes causing myasthe-
c) require complement activation.
nia gravis and Graves’ disease.
d) show induration because of the influx of sensitized T cells and
6. Give an example of an organ-specific ic autoimmune disease and one
macrophages.
that is widespread, involving a varietyty of tissues and organs.
e) depend on activities of the Fc portion of antibodies.
7. Compare and contrast the Arthus reaction
action and serum sickness.
5. Organ transplants, such as of kidneys
8. Why might malnutrition and starvation on lead to immunodeficiencies? a) are experimental at present.
9. What is the most common primary immunodeficiency
mmunodeficiency disorder? b) can be successful only if there are exact matches between donor
10. How can genetic abnormalities leading ng to immunodeficiency disor- and recipient.
ders be corrected? Give an example. c) survive best if radiation is used for immunosuppression.
d) survive best if B cells are suppressed.
Multiple Choice e) are rejected by a complex process in which cellular mechanisms
1. An IgE-mediated allergic reaction predominate.
a) reaches a peak within minutes after
er exposure to antigen. 6. All of the following are true of autoimmune disease except
b) occurs only to polysaccharide antigens.
gens. a) some show association with particular major histocompatibility
c) requires complement activation. types.
d) requires considerable macrophage participation. b) induction of tolerance may alleviate symptoms.
e) is characterized by induration. c) damage to organs occurs due to long-term exaggerated
2. Which of the following statements iss true of the ABO blood group production of IgE.
system in humans? d) disease may result from reaction to viral antigens that are
a) A antigen is present on type O red cells. similar to autoantigens.
b) B antigen is the most common antigen
igen in the population of the e) some are organ-specific and some are widespread in the body.
United States. 7. Autoantibody-induced autoimmune diseases
c) Natural anti-A and anti-B antibodies
es are of the class IgG. a) can sometimes be passively transferred from mother to fetus.
d) People with blood group O do not have natural antibodies against b) include diabetes mellitus.
A and B antigens. c) are always organ-specific.
e) In blood transfusions, incompatibilities
lities cause complement lysis d) are never organ-specific.
of red blood cells. e) cannot be treated.
418 Chapter 17 Immunologic Disorders

8. All of the following approaches are used to treat autoimmune Applications

diseases except 1. Jack and Jill were badly burned in an accident at the well and
a) immunosuppressant drugs. both were taken to the burn unit of the local hospital. The burns
b) induction of tolerance. covered only a small area of skin so grafts were prepared for both
c) antibiotics. patients from the skin of Jack’s thigh. Jack’s graft was successful
d) anti-inflammatory medications. and his burn healed completely. Jill, however, rejected the grafted
e) replacement therapy, as with insulin in diabetes. skin. Explain the immune responses of both patients to these
9. Patients with primary immunodeficiencies in the complement grafts. What treatments could have helped Jill to avoid rejection of
system her graft?
a) who lack late-acting components (C5, C6, C7, C8) show 2. Horse serum containing specific antibody to snake venom has
increased susceptibility to Neisseria infections. been a successful approach to treating snakebite in humans. How
do you think this anti-venom could be generated? What are some
b) who lack C3 are prone to develop tuberculosis.
advantages of using horses to produce the antibody instead of
c) generally have no symptoms.
humans? Why might it be unsafe to administer the anti-venom
d) only show defects in the major components C1 through C9. more than once?
e) usually handle infections normally.
10. One of the most serious of the secondary immunodeficiencies is Critical Thinking +
a) acquired immunodeficiency syndrome, caused by the human 1. Hypersensitivity reactions, by definition, lead to tissue damage.
immunodeficiency virus. Can they also be beneficial? Explain.
b) severe combined immunodeficiency.
2. Explain why people with B-cell deficiencies are more prone to
c) DiGeorge syndrome. bacterial infections, but people with T-cell deficiencies are more
d) chronic granulomatous disease. prone to viral infections.
e) Chediak-Higashi disease.
18 Applications of
Immune Responses
KEY TERMS
Active Immunity Immunity that
results from an immune response in
an individual upon exposure to an
antigen.
antigens on cells attached to a
microscope slide.
Inactivated Vaccine Vaccine
composed of killed bacterial cells,
Adjuvant Substance that increases inactivated virus, or fractions of the
the immune response to antigens. pathogen.
Agglutination Reaction The Passive Immunity Immunity
formation of visible clumps as a that results when antibodies are
result of antibodies binding to and transferred to an individual.
cross-linking insoluble antigens. Precipitation Reaction The
Antiserum A preparation of serum formation of complexes that come out
that contains protective antibodies. of a solution when antibodies bind to
and cross-link soluble antigens.
Attenuated Vaccine Vaccine
composed of a weakened form of the Serology The study of in vitro
pathogen that is generally unable to antibody-antigen reactions; most
cause disease. frequently it implies testing a patient
for specific antibodies to diagnose a
Enzyme-Linked Immunosorbent
disease.
Assay (ELISA) Technique that uses
enzyme-labeled antibodies to detect Western Blotting Procedure that
antigens or antibodies. uses labeled antibodies to detect
specific antigens in a mixture of
Fluorescent Antibody (FA) Test
proteins separated according to their
Technique that uses fluorescently
An immunoassay. molecular weight.
labeled antibodies to detect specific

A Glimpse of History
Long before people knew that microbes caused disease, they recognized hand of a milkmaid, Sarah Nelmes, to a scratch on the arm of a young
that individuals who recovered from a disease such as smallpox rarely boy named James Phipps. Six weeks later, when exposed to pus from a
got it a second time. Old Chinese writings dating from the Sung dynasty smallpox victim, Phipps did not develop the disease. The boy had been
(a.d. 960–1280) describe a procedure known as variolation, in which made immune to smallpox when he was inoculated with pus from the
small amounts of the powdered scabs from smallpox lesions were inhaled cowpox lesion.
or placed into a scratch made in the skin. The resulting disease was usu- Using the less dangerous cowpox material in place of the scabs
ally mild, and the person was then immune to smallpox. Occasionally, from smallpox cases, Jenner and others worked to spread the practice
however, severe disease developed, often resulting in death. In addition, of variolation. Later, Pasteur used the word vaccination (from the Latin
the person became contagious, so the disease could spread. vacca for “cow”) to describe any type of protective inoculation. By the
Although variolation was practiced in China and the Mideast a twentieth century, most of the industrialized world was generally free of
thousand years ago, it was not widely used in Europe until after 1719. At smallpox as the result of routine vaccination.
that time, Lady Mary Wortley Montagu, wife of the British ambassador In 1967, the World Health Organization (WHO) started a program
to Turkey, had their children immunized against smallpox in this way. of intensive smallpox vaccination. Because there were no animal hosts
Variolation then became popular in Europe. Because of the dangers, and no non-immune humans to whom it could be spread, the disease
however, and the fact that the procedure was expensive, many people died out. The last naturally contracted case occurred in Somalia, Africa,
remained unprotected. in 1977, and two years later WHO declared the world free of smallpox.
As an apprentice physician, Edward Jenner noted that milkmaids Nevertheless, a few laboratories around the world still have the virus. In
who had recovered from cowpox (a disease of cows that caused few this age of bioterrorism concerns, some see smallpox as a major threat
or no symptoms in humans) rarely got smallpox. Then, in 1796, long should the deadly virus ever be released into the largely unprotected
before viruses had been discovered, he conducted a classic experiment in populations of the world. Because of this, vaccine stores in the United
which he deliberately transferred material from a cowpox lesion on the States have been increased.

419
420 Chapter 18 Applications of Immune Responses

FIGURE 18.1
The Host- The Immune Wars The Pathogens Fight Back The Return of the Humans
Pathogen Innate Immunity (chapter 14) Pathogenesis (chapter 16) (Knowledge Is Power)
Trilogy Adaptive Immunity (chapter 15) Immunization (chapter 18)
Epidemiology (chapter 19)
? How does Antimicrobial Drugs (chapter 20)
immunization
prevent disease?

C
hapters 14 and 15 discussed the innate and adaptive greatest impact on human health of any medical procedure,
defense systems, describing antibodies and lympho- and it is just one example of how knowledge is power with
cytes. This chapter will consider how immunization, respect to fighting disease (figure 18.1). We will also explore
the process of inducing immunity, can be used to protect some useful applications of immunological reactions in diag-
against disease. In fact, immunization has probably had the nostic tests.

IMMUNIZATION
18.1 ■ Principles of Immunization Artificially acquired passive immunity involves injecting a
person with antibodies produced by other people or animals. This
Learning Outcome can be used to (1) prevent disease immediately before or after
1. Compare and contrast naturally acquired active immunity, likely exposure to a pathogen, (2) limit the duration of certain dis-
artificially acquired active immunity, naturally acquired passive eases, and (3) block the action of microbial toxins. A preparation
immunity, and artificially acquired passive immunity. of serum (the fluid portion of blood that remains after blood clots)
containing the protective antibodies is referred to as antiserum.
Naturally acquired immunity is the acquisition of adaptive One that protects against a given toxin is called an antitoxin.
immunity through normal events, such as exposure to an infec- Two kinds of antisera (or antitoxins) are used.
tious agent. Immunization mimics those same events, protecting Hyperimmune globulin—prepared from the sera of donors
against disease by inducing artificially acquired immunity with high amounts of antibodies to certain disease agents—is
(figure  18.2). The protection provided by immunization can be used to prevent or treat specific diseases. Examples include
either active or passive. tetanus immune globulin (TIG), rabies immune globulin
(RIG), and hepatitis B immune globulin (HBIG). If these
preparations are given during the incubation period, they
Active Immunity
can often prevent severe diseases from developing. Immune
Active immunity is the result of an immune response in an globulin, the IgG fraction of pooled blood plasma from many
individual upon exposure to antigen. Specific B and T cells are donors, has a variety of antibodies due to typical infections
activated and they then proliferate, giving the individual lasting and vaccines experienced by the donors. It is used to protect
protection due to immunological memory. Active immunity can unvaccinated people who have been recently exposed to the
develop either naturally from an actual infection or artificially measles virus and immunosuppressed people who have low
from vaccination. memory, p. 355 levels of antibodies. incubation period, p. 383

Passive Immunity MicroAssessment 18.1

Passive immunity occurs naturally during pregnancy; the moth- Immunity is natural or artificial, active or passive. Active immunity
occurs naturally in response to infections, and artificially in response
er’s IgG antibodies cross the placenta and protect the fetus. These
to vaccination. Passive immunity occurs naturally from maternal
antibodies remain active in the newborn during the first few antibodies transferred during pregnancy and breast feeding, and
months of life, when his or her own immune responses are still artificially through administration of immune serum globulin or
developing. This is why a number of infectious diseases typically hyperimmune globulin.
do not develop until a baby is 3 to 6 months of age, after the 1. How is naturally acquired active immunity different from
maternal antibodies have been degraded. Passive immunity also artificially acquired active immunity?
occurs as a result of breast feeding; the secretory IgA in breast 2. What is antitoxin?
milk protects the digestive tract of the child. Note that passive
3. What would be a primary advantage of passive immunity with
immunity provides no memory; once the transferred antibodies diseases such as tetanus? +
are degraded, the protection is lost.
 Focus Figure
Active Immunity Passive Immunity

Natural Active Immunity Natural Passive Immunity

Immunity that results from an Immunity that results when
immune response in an individual antibodies from a woman are
after exposure to an infectious agent. transferred to her developing fetus
during pregnancy or to an infant
during breast feeding.

Artificial Active Immunity Artificial Passive Immunity

Immunity that results from an Immunity that results when
immune response in an individual antibodies contained in the serum
after vaccination. of other people or animals are
injected into an individual.

FIGURE 18.2 Acquired Immunity Acquired immunity can be natural or artificial, active or passive.
? Why does active immunity last longer than passive immunity?

18.2 ■ Vaccines and Immunization developing countries. Unfortunately, we periodically see some
of these diseases reappear and spread as a direct consequence of
Procedures parents failing to have their children vaccinated. Table 18.1 lists
a number of human diseases for which vaccines are available. As
Learning Outcomes the table indicates, some are routinely given, whereas others are
2. Compare and contrast the characteristics of attenuated and used only in special circumstances.
inactivated vaccines. Effective vaccines should be safe, with few side effects,
3. List six diseases that routine childhood immunizations have while giving lasting protection against the illness. They should
reduced in occurrence by at least 95%. induce protective antibodies or immune cells, or both, as appro-
priate. For example, polio vaccine should induce antibodies that
A vaccine is a preparation of a pathogen or its products used to neutralize the virus, thereby preventing it from reaching and
induce active immunity. Vaccines not only protect an individual attaching to nerve cells to cause the paralysis of severe polio-
against disease, they can also prevent diseases from spreading in a myelitis. In contrast, an effective vaccine against tuberculosis
population. This is because a phenomenon called herd immunity would induce cell-mediated immunity that can limit growth of
develops when a critical portion of a population is immune to the intracellular bacterium. Of course, vaccines ideally should be
a disease, either through natural immunity or vaccination. The low in cost, stable with a long shelf life, and easy to administer.
infectious agent is unable to spread because there are not enough cell-mediated immunity, p. 356
susceptible hosts. Herd immunity is responsible for dramatic Vaccines fall into two general categories—attenuated and
declines in childhood diseases, both in the United States and in inactivated, based on whether or not the immunizing agent can
TABLE 18.1 Some Important Vaccines for Humans
Disease or
Infectious Agent Type of Vaccine Persons Who Should Receive the Vaccine

Anthrax Subunit Adults in occupations that put them at risk of exposure, such as military
personnel
Diphtheria Toxoid Children (the “D” in the DTaP vaccine given to children); adolescents and adults
receive a booster every 10 years (the “d” in the Td and Tdap booster vaccines)
Haemophilus Polysaccharide-protein conjugate Children
influenzae type b
infections
Hepatitis A Inactivated virus Children; adolescents who live in selected areas; adults with indications that
put them at increased risk (such as traveling to certain countries; men who
have sex with men); close contacts of internationally adopted children.
Hepatitis B Subunit Newborns and children; also adults with indications that put them at
increased risk (such as healthcare workers who might be exposed to blood,
people who have multiple sexual partners, and contacts of infected people)
Human VLP (two or four serotypes) Girls/women ages 11–26
papillomavirus
(HPV) infection
Influenza Three age-specific forms—inactivated virus All people 6 months of age or older; given yearly, as the antigens of
(TIV), hemagglutinin antigen (Fluzone High the virus change frequently. When the vaccine supply is limited, then
Dose), and attenuated virus (LIAV); the first vaccination should focus on high-risk groups
two are given by injection and the latter as
a nasal mist
Measles Attenuated virus Children (an “M” in the MMRV or MMR vaccine given to children);
booster(s) for adults born after 1956 who do not have evidence of
immunity and do not have a medical contraindication
Meningococcal Two age-specific forms active against Adolescents; also children and adults with certain medical conditions that
disease four serotypes—meningococcal conjugate put them at greater risk (for example, those without a spleen or who have
vaccine (MCV4 and MenACWY-CRM) and certain complement system defects); also adults in certain high-risk groups
meningococcal polysaccharide vaccine (such as college students living in dormitories and people traveling to
(MPSV4) sub-Saharan Africa)
Mumps Attenuated virus Children (an “M” in the MMRV or MMR vaccine given to children);
booster(s) for adults born after 1956 who do not have evidence of
immunity and do not have a medical contraindication
Pertussis Subunit (acellular vaccine) Children (the “aP” in the DTaP vaccine given to children); adolescents
(whooping cough) should receive a booster (the “ap” in the Tdap booster vaccine); adults
younger than age 65 may receive a booster
Pneumococcal Two age-specific forms—purified Children; adults age 65 and over, people with certain chronic infections,
infection polysaccharide (PPV) and polysaccharide and others in high-risk groups
protein conjugate (PCV)
Polio Two forms—inactivated virus (Salk vaccine) Children; attenuated virus is used for global control
and attenuated virus (Sabin vaccine)
Rabies Inactivated virus People exposed to the virus, people at high risk for exposure, such as
veterinarians and other animal handlers
Rotavirus infection Attenuated virus Children
Rubella (German Attenuated virus Children (the “R” in the MMR or MMRV vaccine given to children);
measles) women who do not have evidence of immunity and do not have a medical
contraindication
Shingles Attenuated virus Adults age 60 and over
Tetanus Toxoid Children (the “T” in the DTaP vaccine given to children); adults receive a
booster every 10 years (the “T” in the Td and Tdap vaccines)
Tuberculosis Attenuated bacterium (BCG strain) Used only in special circumstances in the United States; widely used in other
countries
Typhoid fever Two forms—attenuated bacterium People traveling to certain parts of the world
(Ty21a strain; taken orally) and purified
polysaccharide (ViCPS)
Varicella-zoster Attenuated virus Children (“V” in the MMRV vaccine given to children); also adults without
(chickenpox) evidence of immunity
Yellow fever Attenuated virus Travelers to endemic areas
 Part III Microorganisms and Humans 423

TABLE 18.2 A Comparison of Characteristics of Attenuated and Inactivated Vaccines

Characteristic Attenuated Vaccines Inactivated Vaccines

Antibody response (memory) IgG; secretory IgA if administered orally or nasally IgG
Cell-mediated immune response Good Poor
Duration of protection Long-term Short-term
Need for adjuvant No Yes
Number of doses Usually single Multiple
Risk of mutation to virulence Very low Absent
Risk to immunocompromised recipient Can be significant Absent
Route of administration Injection, oral, or nasal Injection
Stability in warm temperatures Poor Good
Types Attenuated viruses, attenuated bacteria Inactivated whole agents, toxoids, subunit
vaccines, VLPs, polysaccharide vaccines

replicate. Each type has characteristic advantages and disadvan- developing countries where they are desperately needed, is that
tages (table 18.2). they usually require refrigeration to keep them active.
Attenuated vaccines currently in widespread use include those
Attenuated Vaccines against measles, mumps, rubella, chickenpox, rotavirus, and yellow
fever. The Sabin vaccine against polio is also an attenuated vaccine.
An attenuated vaccine is a weakened form of the pathogen that
generally cannot cause disease. The attenuated strain replicates
in the vaccine recipient, causing an infection with undetectable Inactivated Vaccines
or mild disease that typically results in long-lasting immunity. An inactivated vaccine is unable to replicate, but retains the immuno-
Because infection with the attenuated strain mimics that of the genicity of the pathogen or toxin. The advantage of inactivated vac-
wild-type strain, it evokes the type of immune response appropri- cines is that they cannot cause infections or revert to pathogenic forms.
ate for controlling the infection. For instance, attenuated vaccines Because they do not replicate, however, there is no amplification of
given orally induce mucosal immunity (a secretory IgA response), the dose in vivo, so the magnitude of the immune response is limited.
protecting against pathogens that infect via the gastrointestinal To compensate for the relatively low effective dose, several booster
tract. Some attenuated vaccines are able to stimulate cytotoxic doses are usually needed to induce sufficient immunity to be protec-
T cells, inducing cell-mediated immunity. mucosal immunity, p. 358 tive. Some inactivated vaccines include the whole infectious agent,
Attenuated strains are often produced by growing a microbe under and others include only fractions of the agent. Examples include:
conditions that cause mutations to accumulate, making the microbe
■ Inactivated whole agent vaccines. These contain killed
less pathogenic. Viruses of humans can sometimes be attenuated by
microorganisms or inactivated viruses. The vaccines are made
growing them in cells of a different animal species; the mutations that
by treating the pathogen with formalin or another chemical
allow them to multiply in the other animal cells often cause them to
that does not significantly change the surface epitopes. The
grow poorly in human cells. Genetic manipulation is now being used
treatment leaves the agent immunogenic even though it can-
to produce strains of pathogens with low virulence. Specific genes
not reproduce. Vaccines in this category include those against
are mutated and used to replace wild-type genes. The inserted mutant
influenza, rabies, and polio (Salk vaccine). formalin, p. 118
genes are engineered so they cannot revert to the wild type.
Attenuated vaccines have several advantages compared to ■ Toxoids. These are inactivated toxins used to protect against
their inactivated counterparts. For one thing, a single dose of an diseases caused by bacterial toxins. They are prepared by
attenuated agent is often enough to induce long-lasting immunity. treating the toxins to destroy the toxic part of the molecules
This is because the microbe multiplies in the body, causing the while retaining the antigenic epitopes. Diphtheria and tetanus
immune system to be exposed to the antigen for a longer period vaccines are toxoids.
and in greater amounts than with inactivated agents. In addition, ■ Subunit vaccines. These consist of key protein antigens
the vaccine strain has the added potential of being spread from an or antigenic fragments of a pathogen. Obviously, they can
individual being immunized to other non-immune people, inad- be developed only after research has revealed which of the
vertently immunizing the contacts of the vaccine recipient. microbe’s components are most important in triggering a
The disadvantage of attenuated agents is they sometimes cause protective immune response. Their advantage is that cell parts
disease in immunosuppressed people, and can occasionally mutate that may cause undesirable side effects are not included. The
to become pathogenic again. Attenuated vaccines are generally vaccine currently used to prevent whooping cough (pertus-
not advised for pregnant women because of the possibility that the sis) is a subunit vaccine, referred to as the acellular pertussis
vaccine strain may cross the placenta and damage the developing (aP) vaccine. It does not cause the side effects that sometimes
fetus. Another disadvantage of attenuated vaccines, especially in occurred with the killed whole-cell vaccine used previously.
424 Chapter 18 Applications of Immune Responses

■ Recombinant vaccines. These are subunit vaccines produced intestine, but the vaccine still has to be given in a series of three
by genetically engineered microorganisms. An example is the doses because of interactions among the viruses.
vaccine against the hepatitis B virus; it is produced by yeast Attenuated and inactivated polio vaccines both cause the
cells engineered to produce part of the viral protein coat. immune system to produce antibodies that protect against viral
■ VLP (virus-like particle) vaccines. These are empty capsids. invasion of the central nervous system and consequent paralytic
Laboratory organisms are genetically engineered to produce poliomyelitis. The Sabin vaccine, however, has a distinct advan-
the major capsid proteins of a virus, which then self-assemble. tage over the Salk vaccine in that it induces better mucosal immu-
The human papillomavirus (HPV) vaccines are VLPs. nity (secretory IgA response), and therefore provides better herd
immunity. A disadvantage of the Sabin vaccine is that the attenu-
■ Polysaccharide vaccines. These contain the polysaccharides
ated viruses can mutate to become virulent. Approximately one
that make up the capsules of certain organisms. They are
case of poliomyelitis arises for every 2.4 million doses of Sabin
not effective in young children because polysaccharides are
vaccine administered. poliomyelitis, p. 656
T-independent antigens; recall that these antigens generally
An obvious way to avoid vaccine-related poliomyelitis is to
elicit a poor response in this age group. The pneumococcus
abandon the Sabin vaccine in favor of the Salk. As usual, however,
vaccines given to adults are polysaccharide vaccines.
the situation is not as simple as it might seem. The Sabin vaccine,
■ Conjugate vaccines. These are polysaccharides linked to pro- unlike the Salk vaccine, provides better protection against trans-
teins, a modification that converts the polysaccharides into mission of the wild-type virus. If only the inactivated vaccine is
T-dependent antigens. The first conjugate vaccine developed was used, the virus can still replicate in the gastrointestinal tract and be
against Haemophilus influenzae type b (Hib) and has nearly elim- transmitted to others, rapidly spreading in a population.
inated Hib meningitis in children. The conjugate vaccine devel- A campaign to eliminate polio using the Sabin vaccine was
oped against certain Streptococcus pneumoniae strains promises so successful that by 1980 the United States was free of wild-type
to do the same for a variety of infections caused by those strains. polioviruses (see figure  26.14). By 1991, the viruses had been
Haemophilus influenzae type b, p. 647 Streptococcus pneumoniae, p. 497 eliminated from the Western Hemisphere. Because of the contin-
ued risk of vaccine-associated paralytic polio, a vaccine strategy
Many inactivated vaccines contain an adjuvant, a substance that attempted to capture the best of both vaccines was adopted in
that enhances the immune response to antigens (adjuvare means the United States. Children first received doses of the Salk vac-
“to help”). These are necessary additives because purified antigens cine, protecting them from poliomyelitis. Following these doses,
such as toxoids and subunit vaccines are often poorly immunogenic the Sabin vaccine was given, providing mucosal protection while
by themselves because they lack the “danger” signals—the patterns also boosting immunity. The routine use of the Sabin vaccine was
associated with tissue damage or invading microbes. These patterns then discontinued altogether in the United States.
cause dendritic cells to produce co-stimulatory molecules, allowing The original goal of global eradication of polio by 2000 was
them to activate helper T cells, which, in turn, activate B cells not achieved, but efforts continue. For these eradication programs,
(see figure 15.20). Adjuvants are thought to function by providing the Sabin vaccine must be used because it prevents transmission
the danger signals to dendritic cells. Some adjuvants appear to of the virus.
adsorb the antigen, releasing it at a slow but constant rate to the
tissues and surrounding blood vessels. Unfortunately, many effec- The Importance of Childhood
tive adjuvants trigger an intense inflammatory response, making
them unsuitable for use in vaccines for humans. Alum (aluminum
Immunizations
hydroxide and aluminum phosphate) is the most common adjuvant Before vaccination was available for common childhood diseases,
used, but others, including one that uses a derivative of lipid A, thousands died or were permanently disabled from these diseases
have recently been developed. pattern recognition, p. 342 dendritic (table 18.3). Unfortunately, many people still become ill or even
cells, pp. 340, 370 lipid A, p. 60 die from diseases easily prevented by vaccines.
One reason some children are not protected is that their par-
An Example of Vaccination ents have refused to have them vaccinated, fearing that vaccina-
tion might be harmful. In situations such as this, vaccines have
Strategy—The Campaign become victims of their own success. They have been so effective
to Eliminate Poliomyelitis at preventing diseases that people have been lulled into a false
Vaccines against poliomyelitis provide an excellent illustration sense of security. Reports of adverse effects of vaccination have
of the complexity of vaccination strategies. The virus that causes led some people to falsely believe that the risk of vaccination is
this disease enters the body orally, infects the throat and intestinal greater than the risk of diseases.
tract, and then invades the bloodstream. From there, it can invade There will always be at least some risk associated with almost
nerve cells and cause the disease poliomyelitis (see figure 26.13). any medical procedure, but there is no question that the benefits of
There are three types of poliovirus, any of which can cause polio- routine vaccinations greatly outweigh the very slight risks. Data
myelitis. The Salk vaccine, developed in the mid-1950s, consists show that a child with measles has a 1:2,000 chance of developing
of inactivated viruses of all three types. It successfully lowered serious brain inflammation, compared with a 1:1,000,000 chance
the rate of the disease dramatically but had the disadvantage of from the measles vaccine. Between 1989 and 1991 measles
requiring a series of injections for maximum protection. In 1961, immunization rates dropped 10% and an outbreak of 55,000 cases
the Sabin vaccine became available, with the advantage of cheaper occurred, with 120 deaths. Now that the vaccination rates have
oral administration. The attenuated vaccine strains replicate in the increased again, measles outbreaks are rarely seen.
 Part III Microorganisms and Humans 425

Vaccines in the CDC recommended immunization

The Effectiveness of Universal
TABLE 18.3 schedule are generally covered by the National Vaccine
Immunization in the United States
Injury Compensation Program. This no-fault alterna-
Cases per Year Decrease After tive for resolving vaccine injury claims was established
Disease Before Immunization Immunization to stabilize the U.S. vaccine market. An excise tax on
Smallpox 48,164 (1900–1904) 100%
every vaccine dose purchased funds the program.
Diphtheria 175,885 (1920–1922) Nearly 100%
Pertussis (whooping cough) 147,271 (1922–1925) 93.4% Current Progress in Immunization
Tetanus 1,314 (1922–1926) 98.1% Advances in understanding the immune system allow
Paralytic poliomyelitis 16,316 (1951–1954) 100% researchers to make safer and more effective vaccines.
An excellent example of this is the introduction of con-
Measles 503,282 (1958–1962) Nearly 100%
jugate vaccines designed to enlist T-cell help. Another
Mumps 152,209 (1968) 99.8%
is the new adjuvants being developed based on insights
Rubella (German measles) 50,230 (1966–1969) 98% gained from the discovery of toll-like receptors (TLRs).
Haemophilus influenzae 20,000 (estimated) 99.8% The immune response can also be improved by admin-
type b invasive disease in istering certain cytokines with vaccines to guide that
children
response. TLRs, p. 342
Novel types of vaccines being studied include pep-
Routine immunization against pertussis (whooping cough) tide vaccines, edible vaccines, and DNA-based vaccines. None
caused a significant decrease in its incidence in the United States of these rely on whole cells, so they eliminate the possibility of
and saved many lives. Because of some adverse reactions to the infection with the immunizing agent, but some are only weakly
killed whole-cell vaccine being used at the time, however, many immunogenic. Peptide vaccines are composed of key antigenic
parents refused to allow their babies to get the vaccine. By 1990, peptides from disease-causing organisms. They are stable to heat
this resulted in the highest incidence of pertussis in 20 years and and do not contain extra materials that might cause unwanted
the deaths of some children, mostly those under one year of age. reactions or side effects. Edible vaccines are created by transfer-
An acellular subunit pertussis vaccine, which is more effective and ring genes encoding key antigens from infectious agents into
has fewer side effects than the whole-cell vaccine, is now used. plants. If appropriate plants can be genetically engineered to
The suggestion that vaccines are associated with autism has function as vaccines, they could potentially be grown throughout
again threatened the acceptance of immunization. It is important the world, eliminating difficulties involving transport and storage.
to note, however, that scientific studies show no evidence of a link DNA-based vaccines are segments of naked DNA from infectious
between the two. organisms that can be introduced directly into muscle tissue. The
The U.S. Centers for Disease Control and Prevention (CDC) host tissue actually expresses the DNA for a short period of time,
regularly publishes recommended immunization schedules for producing the encoded microbial antigens, which induces an
children, adolescents, and adults. These are updated regularly as immune response.
vaccines are developed and modified. Because of the complex- There are several serious and widespread diseases for which
ity of the schedules and the frequency at which they have been new or more effective vaccines are currently being sought
updated recently, it is important to know how to access the most (table  18.4). Many of the pathogens involved are quite good at
current schedule. A link can be found in the chapter 18 readings avoiding the host defenses, complicating the development of long-
at the text website (www.mhhe.com/nester7). lasting effective vaccines. In addition to seeking vaccines that
protect against infectious diseases, other uses of vaccines are also
being studied, including vaccines to treat cancer.
Some Diseases for Which New
TABLE 18.4
or Improved Vaccines Are Sought
MicroAssessment 18.2
Disease Estimated Impact
An attenuated vaccine is a weakened form of the pathogen.
HIV/AIDS 40 million infected worldwide, with An inactivated vaccine is unable to replicate but retains the
approximately 14,000 new infections daily immunogenicity of the pathogen or toxin; examples include killed
Malaria 300–500 million cases/yr and up to 3 million
microorganisms, inactivated viruses, and fractions of the agents,
deaths/yr worldwide including toxoids. Routine childhood immunizations have prevented
millions of cases of disease and many deaths during the past decades.
Influenza 30–50 million cases/yr worldwide; 10,000–40,000 Many experimental vaccines are under study or in clinical trials.
deaths/yr in the United States
4. What is the difference between an attenuated and inactivated
Strep throat 20 million cases/yr in the United States
vaccine?
Genital herpes 45 million infected and 500,000 new infections/yr 5. Childhood diseases such as measles and mumps are rare now, so
in the United States
why is it important for children to be immunized against them?
Hepatitis C 170 million infected worldwide 6. What would be a primary advantage of using an attenuated
Cancer 1 in 3 in the United States may get cancer, agent for a vaccine rather than just an antigen from that
resulting in 560,000 deaths/yr agent? +
426 Chapter 18 Applications of Immune Responses

IMMUNOLOGICAL TESTING
Immunoassays take advantage of the specificity of antibody- Just as the field of immunology has advanced significantly
antigen interactions, using them for diagnosis. For example, if in the last few decades, so has immunological testing. New tests
a person is suspected of having syphilis, antibodies that bind are continually being developed, supplementing or gradually
specifically to the causative bacterium, Treponema pallidum, can replacing many of the older methods. This section will focus
be added to fluid collected from a genital lesion. If the antibodies primarily on tests commonly used today; information about other
bind to a bacterium in the specimen, they identify the organism as tests—including radial immunodiffusion, immunoelectrophoresis,
T. pallidum, indicating that the patient does indeed have syphilis complement fixation, radioimmunoassay and hemagglutination
(figure 18.3). In addition to identifying microbes, immunoassays inhibition—which are declining in use but illustrate key immuno-
can be used to detect specific antibodies. If a patient’s blood has logical principles, can be found in the chapter 18 readings at the
antibodies to a given microbe, then the patient’s immune system text website (www.mhhe.com/nester7).
must have responded to the microbe at some point, indicating
either previous or current infection.
One of the earliest examples of immunological testing is
the tuberculin skin test (also called the Mantoux test), which 18.3 ■ Principles of
is still used for diagnosing tuberculosis. People infected with Immunological Testing
Mycobacterium tuberculosis develop a strong cell-mediated
response to the bacterium and its products, which is the basis Learning Outcomes
of the test. When a purified protein derivative (PPD) from 4. Describe the difference between polyclonal and monoclonal
M. tuberculosis is injected into the skin of someone infected with antibodies.
the organism, redness and a firm swelling usually develop at 5. Describe how the antibody titer is determined.
the site (see figure  17.6). In contrast, people who have not been
infected show little, if any, response. A person who has not been exposed to a given pathogen typi-
cally lacks specific antibodies against the microbe in their serum,
and is referred to as seronegative. Once infected, that person will
begin producing specific antibodies about a week to 10 days later,
An unknown Solution containing becoming seropositive. This change from seronegative to sero-
organism known antibodies to positive is referred to as seroconversion. As the infection pro-
Treponema pallidum gresses, increasing amounts of specific antibodies are produced.
Binding of known
A rise in the concentration (titer) of specific antibodies is charac-
antibodies identifies teristic of an active infection. In contrast, small but steady levels
bacterium as of specific antibodies indicate a previous infection or vaccination.
+ Treponema pallidum.
To determine if a person has specific antibodies in the blood,
then either the serum or plasma is tested. Serum is the fluid por-
tion of blood that remains after blood clots; plasma is the fluid
portion of blood treated with an anticoagulant to prevent clot-
ting. Because serum is so often used as a source of antibodies,
(a)
the study of in vitro antibody-antigen interactions is referred
to as serology. Most frequently it implies testing a patient for
Antibodies of Known specific antibodies to diagnose a disease. Cerebrospinal fluid,
unknown specificity Treponema
in a patient’s serum pallidum tissues, and other clinical specimens can also be tested for
antibodies.
Binding of antibodies in
patient’s serum to
known Treponema Obtaining Antibodies
+
pallidum suggests past
or current infection.
Several different methods are used to obtain antibodies. The
approach chosen depends largely on the intended use of the product.

Polyclonal Antibodies
(b) Laboratory animals are used to produce antibodies known to bind
FIGURE 18.3 Principles of Immunoassays These assays can be a certain infectious agent. The animals are immunized with either
used to (a) identify unknown bacteria (or other antigens); (b) detect the whole agent or part of the agent, and the resulting antibodies
specific antibodies. are then collected by harvesting the animal’s serum. The anti-
? What three occurrences could account for a person having body preparation will be polyclonal, meaning that multiple naive
antibodies to a specific infectious agent? B cells responded to the immunization, giving rise to a mixture
 Part III Microorganisms and Humans 427

of antibodies that together recognize a variety of epitopes on the

antigen. The more complex the antigen, the greater the number of
different epitopes recognized by the polyclonal antibodies. For
instance, injecting whole bacteria will result in a wider array of
antibody specificities than injecting purified toxin.
One problem with polyclonal antibodies is that some may
bind to closely related organisms, resulting in a false positive
reaction. As an example, Shigella species have outer membrane
proteins in common with E. coli, so an animal immunized with
whole Shigella cells would produce some antibodies that also bind
E. coli cells. If those antibodies were used in a diagnostic test for
Shigella, a specimen containing E. coli but not Shigella would
yield a false positive result.
Certain serological tests discussed in this chapter use anti-
bodies that bind to the constant region of human IgG molecules.
These are referred to as anti-human IgG antibodies. They are FIGURE 18.4 Microtiter Plate Serological tests can be done in
produced by animals that have been immunized with IgG from the wells of these small plates.
human serum. Anti-human IgG antibodies are available com- ? What is the advantage of doing serological tests in a microtiter
mercially as are antibodies that bind to the other immunoglobulin plate?
classes.

Monoclonal Antibodies Serology tests can be done in test tubes, but this requires
Monoclonal antibodies recognize only a single epitope. They many tubes and large amounts of reagents. Therefore, the tests are
are difficult and expensive to develop, so a given specificity is usually done using plastic microtiter plates, which have 96, 384,
generally available only if it has commercial value due to wide- or 768 wells (figure 18.4). The volumes used in each well are a
spread use. mere fraction of those needed for even a small test tube, so tests
Monoclonal antibodies are obtained through a complicated can be done on very small samples. Special equipment can be used
process that involves taking B cells from an immunized animal, to mix the reagents and read the results.
and then fusing those short-lived B cells with other cells that will
divide repeatedly in culture (see Perspective 18.1). Because each MicroAssessment 18.3
B cell is programmed to produce antibody molecules that recog-
Antibodies used in serological tests can be either polyclonal or
nize only a single epitope, antibody preparations produced by the
monoclonal. Serial dilution of serum or plasma permits quantification
descendants (clones) of a single B cell are all identical. of antibodies in the specimen.
A monoclonal antibody derived from a mouse or other labo-
7. What is the significance of a rise in titer of specific antibodies in
ratory animal can be “humanized.” To do this, recombinant DNA
serum samples taken at different times?
techniques are used to replace most of the antibody molecule with
8. How are polyclonal antibodies different from monoclonal
the human equivalents. This gives the molecule a longer half-
antibodies?
life in humans because the human immune system is less apt to
9. Would antibodies produced by a patient in response to
destroy it. Some humanized monoclonal antibodies are given as a
infection be monoclonal or polyclonal? +
form of passive immunity to treat certain types of cancers. Others
are tagged with a drug or toxic substance and then used to deliver
that tag to a specific cell type in vivo. These tagged monoclonal
antibodies have been used to treat non-Hodgkin’s lymphoma that 18.4 ■ Observing Antigen-
has not responded to traditional treatment. Antibody Aggregates
Learning Outcome
Quantifying Antigen-Antibody 6. Compare and contrast precipitation reactions and agglutination
Reactions reactions.

The concentration of antibody molecules in a specimen such as

Recall from chapter 15 that antibodies can cross-link antigens,
serum is usually determined by making serial dilutions similar to
creating large “mouthfuls” for phagocytic cells (see figure 15.8).
those done to count bacterial cells (see figure  4.17). Sequential
This type of antigen-antibody interaction can be observed in pre-
two-fold or ten-fold dilutions are used to dilute the specimen,
cipitation and agglutination reactions.
and then antigen is added to each dilution. The titer (concentra-
tion) is expressed as the reciprocal of the last dilution that gives a
detectable antigen-antibody reaction. Thus, if a positive reaction is Precipitation Reactions
observed in the dilution 1:256 but not in 1:512, then the antibody When antibodies bind to soluble antigens, the molecules
titer is 256. sometimes cross-link to form latticelike insoluble complexes
428 Chapter 18 Applications of Immune Responses

PERSPECTIVE 18.1
Monoclonal Antibodies

Myeloma cells. These abnormal

Immunize a mouse with B cells from spleen. These are plasma cells grow indefinitely,
antigen X to activate and capable of making anti-X antibodies, cannot make antibodies, and have
induce proliferation of but die after several generations. a mutation that makes them
specific B cells.
susceptible to the drug aminopterin.
FIGURE 1 Production of
Monoclonal Antibodies

When an animal is injected with an

antigen, its immune system responds
by making antibodies directed against Mix the two cell types along
B cells die. Myeloma cells die. with a chemical that induces
the antigen’s different epitopes. Even their fusion, and then incubate
though there is a single antigen, a in a medium that contains
variety of different B cells respond, aminopterin. The B cells and
resulting in the production of poly- myelomas die, but hybridomas
proliferate.
clonal antibodies. Unfortunately,
this makes it difficult to standardize
experimental results, because the anti-
body composition is different each Hybridoma cells. These are fusions
time the antiserum is made. of B cells and myeloma cells.
In 1975, Georges Köhler and
Cesar Milstein overcame the prob-
lem of variable antiserum prepara-
tions by developing a technique to
make monoclonal antibodies. These
are antibodies produced by a single Select single hybridoma cell
B clone, so all molecules in a prepa- that recognizes desired anti-X
epitope and maintain it in
ration will have the same constant culture.
and variable regions and, thus, the
same functional characteristics and
specificity. With such consistency,
tests can be standardized more
easily and with greater reliability.
To make monoclonal antibodies,
a laboratory animal is immunized
with the agent being studied, and Harvest antibodies made by the
hybridoma cells.
B lymphocytes are then isolated.
These are then fused with myeloma
cells, which are malignant (cancer- Monoclonal antibodies. These all have the same constant
ous) plasma cells. Unlike normal and variable regions, and therefore recognize the same
plasma cells, these myeloma cells epitope and have the same functional characteristics.
can divide repeatedly in culture, do
not make antibodies, and have lost
the capacity to produce a critical enzyme, so traits from the fused cells: The B cell supplies For example, monoclonal antibodies against
they cannot grow in a medium that contains the genes for the specific antibody production, a hormone can detect pregnancy only 10 days
the drug aminopterin. When the B cells and and the myeloma cell supplies the cellular after conception. Specific monoclonal antibod-
myeloma cells are mixed and grown in a machinery for producing the antibodies and ies are used for rapid diagnosis of hepatitis,
medium that contains aminopterin, only the multiplying indefinitely. influenza, herpes simplex, and Chlamydia
fusion products, called hybridomas, can pro- In the laboratory, monoclonal antibodies infections. Köhler and Milstein won the Nobel
liferate. The hybridoma cells retains critical are the basis of a number of diagnostic tests. Prize in 1984 for their work.
 Part III Microorganisms and Humans 429

Zone of Zone of that then precipitate out of solution (figure 18.5). This is the
antibody excess antigen excess
basis of precipitation reactions, which can be used to detect
specific antibodies or antigens. The complexes (aggregates)
Amount of antibody precipitated

Zone of can take several hours to form, and develop only at certain
optimal relative concentrations of antibody and antigen molecules. If
proportions there is a great excess of either, the aggregates do not form,
and consequently, no precipitate will be seen. The easiest way
to get the proper concentration is to place the antigen and
antibody suspensions near each other in a gel, and let the mol-
ecules diffuse toward each other. A precipitate will form in a
distinct region called the zone of optimal proportions.
The use of precipitation reactions in diagnosis has largely
been replaced by methods that will be discussed shortly.
However, the principle of the reactions is nicely demonstrated
Relative amount of antigen by the Ouchterlony technique, which can be done in a Petri
dish (figure 18.6). Antigen and antibody solutions are placed
Antibody Soluble
antigen into separate wells cut in the gel contained in the dish. These
solutions will gradually diffuse outward, meeting between
the wells. If the antibody molecules recognize the antigen,
a line of precipitation will form at the zone of optimal pro-
portions. Because there are often multiple antigens present
in the sample, as well as different specificities of antibodies
in the serum, more than one line can form, each in its area
of optimal proportions. The Ouchterlony test can be used
In the zone of antibody In the zone of optimal In the zone of antigen
excess, little or no proportions, extensive excess, little or no to detect autoantibodies associated with certain connective
cross-linking occurs; cross-linking occurs; a cross-linking occurs; tissue disorders. Other immunodiffusion tests (precipitation
no visible precipitate visible precipitate forms. no visible precipitate tests carried out in a gel) are described on the text website
forms. forms.
(www.mhhe.com/nester7). autoantibody, p. 412
FIGURE 18.5 Antigen-Antibody Precipitation Reactions
The maximum amount of precipitate forms in the zone of optimal
proportion.
? It takes fewer molecules of IgM than IgG to cause precipitation.
Why would this be so?

Positive Anti-X Antibody and antigen solutions are placed

control antibodies into separate wells cut into the gel. The
antigens and antibodies diffuse toward each
1 other. In this example, antibodies that bind
6
antigen X (called anti-X antibodies) were
added to the well in the center of the gel.
2 Samples that contain unknown antigens are
5 added to wells 1 through 4. A negative
control (contains no antigen X) was added
to well 5, and a positive control (contains
3 known antigen X) was added to well 6.
4
Negative
control
When antibody molecules that recognize
the antigen meet at the zone of optimal
6 1 proportions, antigen-antibody complexes
precipitate out of solution, forming a visible
line. In this example, a line has formed
2 between the center well and well 6 (the
5 positive control). A line has also formed
between the center well and the sample in
3 well 4, indicating that the sample contains
4 antigen X. The other samples do not
contain detectable amounts of antigen X. (b)
FIGURE 18.6 Ouchterlony Technique (a) Method.
(b) Photograph of results.
(a) ? What is the purpose of including positive and negative controls?
430 Chapter 18 Applications of Immune Responses

Anti-A Anti-B
used to test for various bacteria, fungi, viruses, and parasites, as
well as hormones, drugs, and other substances. The beads are
mixed with a drop of a body fluid or suspended microbial cul-
ture. If the specific antigen is present, visible clumps form. Latex
agglutination tests are commonly used to identify beta-hemolytic
Streptococcus species (figure 18.8).

MicroAssessment 18.4
Agglutination and precipitation reactions both depend on cross-
linking and lattice formation of antigen-antibody complexes.
(a) (b) 10. How do the antigens used in precipitation reactions differ from
those in direct agglutination tests?
FIGURE 18.7 ABO Blood Typing This method tests for two 11. How is a direct agglutination test different from a passive
different antigens (A and B) on red blood cells. (a) The red blood agglutination test?
cells agglutinated when mixed with anti-A antibodies. (b) The red 12. In precipitation reactions, why can cross-linked lattices not form
blood cells did not agglutinate when mixed with anti-B antibodies. when there is an excess of antibody? +
Together, the results in panels (a) and (b) indicate that the blood
group is type A.
? What term is used to describe the agglutination of red blood cells?
18.5 ■ Using Labeled Antibodies
Agglutination Reactions to Detect Antigen-
Agglutination and precipitation reactions are similar in principle—
Antibody Interactions
both depend on cross-linking and lattice formation. In agglutination
Learning Outcomes
reactions, however, relatively large insoluble particles are involved
7. Explain how labeled antibodies are used in direct and indirect
rather than soluble molecules. Because of this, bulkier aggregates
tests.
form, which are much easier to see.
8. Compare and contrast fluorescent antibody tests, ELISAs, and
In direct agglutination tests, an antibody suspension is mixed
Western blots.
with the insoluble antigen, such as red blood cells, bacteria, or
9. Describe how the fluorescence-activated cell sorter is used in
fungi. If the antibodies bind to the antigens, visible clumping will
immunoassays.
occur—a positive test. The agglutination of red blood cells by
antibody binding or other means is referred to as hemagglutina-
Detectable markers such as enzymes, fluorescent dyes, and radio-
tion, and is used in blood typing (figure 18.7).
active tags can be attached to antibodies, which are then used
Passive agglutination tests use either antibodies or antigens
to detect certain antigens. Marking antigens with fluorescently
attached to particles such as latex beads to make the aggregates
labeled antibodies also provides a mechanism for sorting antigens.
larger and therefore easier to see. Latex beads to which specific
antibodies have been attached are produced commercially and
Basic Principles
Labeled antibodies can be used to identify bacteria or other anti-
gens, or they can be used to detect antibodies of a given specific-
ity. The tests can be either direct or indirect (figure 18.9).
Direct tests are typically used to identify an unknown antigen.
To do this, labeled antibodies of known specificity are added to a
preparation of the antigen attached to a solid surface. For example,
if the antigen is suspected of being “antigen X,” then antibodies
specific to that antigen are added. After a washing step to remove
unbound antibodies, the presence of the labeled antibodies bound
to the antigen identifies the antigen. A variety of antibodies that
bind specifically to common pathogens are commercially avail-
able, and these can be purchased with different detectable labels.
FIGURE 18.8 Latex Agglutination In this example, the latex
Indirect tests are often used to detect antibodies of a given
beads are coated with antibodies that bind specifically to cell wall
antigens of Streptococcus pyogenes. Visible clumping (shown on the specificity in a patient’s serum. The tests are called indirect
left), confirms that the organism is S. pyogenes. Negative test results because they require a labeled secondary antibody to detect the
are shown on the right. first (or primary) antibody. To determine if a serum sample con-
? Why coat latex beads with antibodies rather than simply adding tains antibodies of a given specificity, the serum is added to a
the antibodies to a bacterial suspension? known antigen attached to a solid surface. Any unbound serum
 Part III Microorganisms and Humans 431

Direct Test
Labeled
anti-X
antibody
Detectable
marker Wash off Positive result
unbound Label detected.
antibodies. Test for label. Conclusion:
Antigen X is
Labeled Labeled present.
Unknown Add labeled antibodies antibodies
antigen antibodies bind to antigen. remain.
specific for
antigen X.

Unknown antigen is
attached to a solid Wash off
surface. unbound Negative result
antibodies. Test for label. Label not detected.
Conclusion:
Labeled Labeled Antigen X is
antibodies antibodies not present
do not bind. washed off.

(a)

Indirect Test
Detectable Labeled anti-human
Patient’s Add labeled marker IgG antibody
antibody secondary
(IgG) antibodies
(anti-human IgG);
Wash off wash off Test for Positive result
unbound any that label. Label detected.
antibodies. do not bind. Conclusion:
Patient’s serum
Patient’s Patient’s Labeled secondary has IgG against
Add patient’s antibodies antibodies antibodies bind to antigen X.
Antigen X serum bind to antigen. remain. patient's IgG
(contains and remain.
antibodies). Add labeled
secondary
Antigen X is attached antibodies
to a solid surface. Wash off (anti-human IgG); Test for Negative result
unbound wash off any that label. Label not detected.
antibodies. do not bind. Conclusion:
Patient’s serum
Patient’s Patient’s does not have IgG
No human IgG, against antigen X.
antibodies antibodies present so labeled
do not bind. washed off. secondary antibodies
do not bind.

(b)
FIGURE 18.9 Basic Principles of Tests That Use Labeled Antibodies to Detect Antigen-Antibody Interactions (a) Positive and nega-
tive results of a direct test. (b) Positive and negative results of an indirect test.
? In (b), why would it be more efficient to use labeled anti-human IgG rather than label the patient’s antibodies?

antibodies are then washed off. The secondary antibody is specific antibodies in a variety of different patients. The alterna-
then added, which in this case would be labeled anti-human IgG tive, labeling the antibodies from each patient tested, would be
antibodies. These then bind to any IgG molecules bound to the cost-prohibitive. anti-human IgG antibodies, p. 427
antigen. After another washing step, the presence of the labeled
secondary antibody identifies the primary antibody bound to
the antigen. Note that the usefulness of the labeled secondary Fluorescent Antibody (FA) Test
antibodies is they can bind to any human IgG molecules, regard- The fluorescent antibody (FA) test use fluorescence microscopy
less of the source or specificity. Thus, they can be used to detect to locate fluorescently labeled antibodies bound to antigens fixed
432 Chapter 18 Applications of Immune Responses

Direct Test: Positive Result

Fluorescent Fluorescently
dye molecule labeled antibody
(known specificity)
Add fluorescently
labeled antibodies
of known specificity; View using
Unknown antigen wash off those that fluorescence
do not bind. microscope.

Unknown antigen Slide surface Fluorescently labeled

attached to slide. antibodies bind to antigen.

(a) View using

fluorescence microscope.
Indirect Test: Positive Result
Fluorescent Fluorescently
Antibody
dye molecule labeled
in serum
Add patient’s Add fluorescently anti-human
(IgG)
Known antigen serum; wash off labeled anti-human IgG
antibodies that do IgG; wash off any
not bind. that do not bind.

Known antigen Slide surface Patient’s antibodies bind to antigen. Fluorescently labeled secondary
attached to slide. antibodies bind to IgG.

(b)

FIGURE 18.10 Fluorescent Antibody (FA) Test (a) Positive direct FA test. (b) Positive indirect FA test.
? In this diagram, why is anti-human IgG used in the direct test but not in the indirect test?

to a microscope slide (figure  18.10). The antigens are often developed, including the rapid group A strep tests done in doctors’
bacterial cells, and the antibodies may be bound directly or indi- clinics, and home pregnancy kits (figure 18.12).
rectly. Several different fluorescent dyes, including fluorescein The indirect ELISA is routinely used to test blood and serum
(fluoresces yellow-green) and rhodamine (fluoresces orange-red) for antibodies against HIV. When donated blood is tested, if
can be used to label the antibodies. By using various fluorescent antibodies that bind HIV are found, the blood would not be used
dyes, different antigens in the same preparation can be located. for transfusion. In patient testing, the ELISA is used only as a
fluorescent dyes and tags, p. 49 fluorescence microscopy, p. 44 screen. A complicated but more reliable test such as Western blot-
ting (described next) is then used to confirm the positive ELISA
results.
Enzyme-Linked Immunosorbent
Assay (ELISA) Western Blotting
The enzyme-linked immunosorbent assay (ELISA) uses anti- In the Western blotting technique, the various proteins that make
bodies labeled with an enzyme such as peroxidase from the horse- up an antigen are separated by size before reacting them with anti-
radish plant. The enzyme can be detected using a colorimetric bodies. This makes it possible to determine exactly which proteins
assay, which measures the enzymatic conversion of a colorless the antibodies are recognizing, an essential part of accurate HIV
substrate into a colored product (figure 18.11). The ELISA is testing (figure 18.13).
often done in microtiter plates, making it useful for screening To separate the proteins, a special type of gel electrophoresis
large numbers of specimens. The method is widely used, and a called SDS-PAGE is used. This separates proteins of different
variety of prepared ELISA plates are commercially available. sizes into a series of bands, as smaller proteins move faster than
In direct ELISAs, the antigen is sometimes “captured” by the larger ones. The separated proteins are then transferred to a
antibodies that have been attached to the surface. For example, nylon membrane to immobilize them before a solution of antibody
the wells of ELISA plates used to diagnose giardiasis are coated molecules is added. The steps after that are very similar to those of
with antibodies that bind antigens of the causative agent, Giardia an ELISA. To determine if a patient’s serum has antibodies spe-
lamblia. When a stool sample is added to the well, the antibodies cific for the separated proteins, a sample is added to the blot, and
capture the antigens. An enzyme-labeled antibody is then used to then unbound antibodies are washed off. Enzyme- or radioactively
detect, and therefore identify, the captured antigens. In addition to labeled anti-human IgG antibodies are then added, which attach
standard ELISAs, commercial modifications of the test have been to any antibodies remaining from the serum. Unbound labeled
 Part III Microorganisms and Humans 433

Direct ELISA (Sandwich Method): Positive Result

Enzyme-labeled
Enzyme secondary antibody
Add enzyme-
labeled antibodies
of known Add substrate
specificity; wash that changes color
off any that when acted upon
Add specimen. do not bind. by enzyme.

Antibodies to known antigen Antibodies in the well Enzyme-labeled antibodies Color development
attached to well. “capture” antigen in of known specificity bind indicates a positive result.
specimen. antigen and remain.

(a)

Indirect ELISA: Positive Result

Add enzyme-
labeled anti-
Add patient’s human IgG; Add substrate
serum; wash wash off any that changes
off antibodies antibodies color when acted
that do not bind. that do not bind. upon by enzyme.

Known antigen attached Antibodies in serum bind Enzyme-labeled secondary Color development
to well. to antigen. antibodies bind to IgG and remain. indicates a positive test.

(b)

FIGURE 18.11 Enzyme-Linked Immunosorbent Assay

(ELISA) (a) Positive direct ELISA. (b) Positive indirect ELISA. FIGURE 18.13 Western Blotting
? In the first panel of part (a), what is the purpose of the Results Each strip in this photograph
antibodies attached to the well? contains HIV proteins that have been
gp160 separated by gel electrophoresis and
gp120 then transferred in-place to the strips,
along with a serum control. The dark
bands indicate the locations of bound
antibodies. The results for the strong
p66
reactive control (lane a) show that
p55 antibodies have bound all HIV proteins
gp41 on the strip. The results for the weak
reactive control (lane b) show binding
p39 of antibodies to only certain critical
HIV proteins (gp160/120 and p24). The
results for the non-reactive control
p31 (lane c) show no binding to HIV
proteins.

p24 ? Why are the results of the Western

blot more reliable than those of
the ELISA?

p17

Serum
control

FIGURE 18.12 ELISA Test for Pregnancy The test detects

30

26

31

human chorionic gonadotropin (HCG), an antigen present only in

pregnant women. A urine sample is applied on the left. Two pink a b c
lines indicate reaction of HCG with antibodies, a positive test. A single
line indicates absence of HCG in the urine, a negative test. (a) Strong reactive control
(b) Weak reactive control
? What captures the antigen on the strip? (c) Non-reactive control
434 Chapter 18 Applications of Immune Responses

antibodies are then washed off. Finally, the label is detected, gen- MicroAssessment 18.5
erating a bar code–like pattern that indicates which proteins were
recognized by the patient’s antibodies. Antibodies labeled with a detectable marker can be used to identify
an antigen (direct test) or to detect a patient’s antibodies bound to a
known antigen (indirect test). Examples of methods that use labeled
Fluorescence-Activated Cell Sorter (FACS) antibodies include fluorescence antibody tests, ELISA, and Western
As described in chapter 4, flow cytometry counts cells or other blotting. Fluorescence-activated cell sorters can separate and count
cells labeled with fluorescent antibodies.
particles by measuring the light scattered as they pass single-file
by a laser. A specialized version called the fluorescence-activated 13. What equipment is required for a fluorescent antibody test that
cell sorter (FACS) can be used to count and separate cells labeled is not required for an ELISA?
with fluorescent antibodies. For example, subsets of T cells (CD4+ 14. In HIV testing, why is the Western blot used to confirm ELISA
and CD8+) can be counted and even separated by first mixing the results?
cells with two preparations of monoclonal antibodies (one binds 15. Why is a false positive more significant in HIV testing of
CD4 markers and the other CD8 markers), each labeled with a patients than in screening donated blood for transfusions? +
different fluorescent dye. flow cytometry, p. 99 CD markers, p. 369

FUTURE CHALLENGES 18.1

Global Immunization
In addition to developing new safe and effec- Instead of expensive needles and syringes and in the gastrointestinal mucosa. These are the
tive vaccines, a major challenge for the future painful injections, vaccines may be delivered cells that transfer antigens across the mem-
is delivering available vaccines to populations in a number of easy ways. For example, brane to the Peyer’s patches, where immune
worldwide. When this was done in the case of naked DNA can be coated onto microscopic responses occur. Antigens are incorporated into
smallpox, the disease was eliminated from the gold pellets, which are shot from a gunlike substances known to bind to M cells, thereby
world; and poliomyelitis is almost eradicated apparatus directly through the skin into the making it easier for them to enter M cells and
now. The World Health Organization and the muscle, painlessly. Skin patches deliver anti- Peyer’s patches. M cells, p. 358
governments that support it deserve much gens slowly through the skin. Vaccines against One of the major remaining challenges is
of the credit for these achievements. Much mucous membrane pathogens can be delivered development of an effective vaccine against
remains to be done, however, and gifts totaling by a nasal spray, as in some new influenza HIV that can be administered universally.
well over a billion dollars from Bill Gates, one vaccines, or by mouth. Time-release pills Although anti-HIV vaccines are being stud-
of the founders of Microsoft, and his wife have introduce antigen steadily to give a sustained ied, prospects are dim for a truly effective
stimulated further progress in this area. immune response. immunization program for HIV disease. It is
To immunize universally, it is necessary In addition to sprays and pills that deliver also likely that even if HIV disease is brought
to have vaccines that are easily administered, antigens to mucous membranes in order to under control, a new and currently unforeseen
inexpensive, stable under a variety of envi- induce mucosal immunity, techniques are being challenge will arise during the twenty-first
ronmental conditions, and preferably painless. developed to get antigens directly to M cells century.

Summary
IMMUNIZATION Attenuated Vaccines
An attenuated vaccine is a weakened form of the pathogen that can
18.1 ■ Principles of Immunization (figure 18.2)
replicate but is generally unable to cause disease.
Active Immunity Inactivated Vaccines
Active immunity occurs naturally in response to infections or other Inactivated vaccines are unable to replicate but they retain the immu-
natural exposure to antigens, and artificially in response to vaccination. nogenicity of the infectious agent or toxin. They include inactivated
Passive Immunity whole agents, toxoids, subunits, VLPs, polysaccharides, and conjugates.
Passive immunity occurs naturally during pregnancy and through Adjuvants increase the intensity of the immune response to the antigen
breast feeding, and artificially by transfer of preformed antibodies, as in in a vaccine.
immune globulin and hyperimmune globulin. An Example of Vaccination Strategy—The Campaign
to Eliminate Poliomyelitis
18.2 ■ Vaccines and Immunization Procedures
Both the Sabin and the Salk polio vaccines protect against paralytic
A vaccine is a preparation of a pathogen or its products used to induce
poliomyelitis; the Sabin vaccine induces mucosal immunity.
active immunity. It protects an individual against disease, and can also
provide herd immunity (table 18.1).
 Part III Microorganisms and Humans 435

The Importance of Childhood Immunizations (table 18.3) Agglutination Reactions

Routine childhood immunizations have prevented millions of cases of Agglutination reactions are similar in principle to precipitation reac-
disease and many deaths. tions, but insoluble particles are involved, causing obvious aggregates
to form; examples include direct agglutination tests and passive agglu-
Current Progress in Immunization
tination tests (figures 18.7, 18.8).
Progress in vaccination includes enhancing the immune response to
vaccines and developing new or improved vaccines against certain 18.5 ■ Using Labeled Antibodies to Detect
diseases (table 18.4). Antigen-Antibody Interactions
Basic Principles
IMMUNOLOGICAL TESTING
Tests that use labeled antibodies can be either direct or indirect
18.3 ■ Principles of Immunological Testing (figure 18.9).
A seronegative individual becomes seropositive after initial infection
Fluorescent Antibody (FA) Test
with an agent; this seroconversion takes about a week to 10 days. To
determine if a patient has antibodies in the blood against a specific The fluorescent antibody (FA) test relies on fluorescence microscopy
infectious agent, the patient’s serum or plasma is tested. to locate fluorescently labeled antibodies bound to antigens fixed to a
microscope slide (figure 18.10).
Obtaining Antibodies
Enzyme-Linked Immunosorbent Assay (ELISA)
Polyclonal antibodies recognize multiple epitopes, whereas monoclonal
antibodies recognize only a single epitope. Anti-human IgG antibodies The enzyme-linked immunosorbent assay (ELISA) uses antibodies
can detect IgG molecules in a patient specimen. Recombinant labeled with a detectable enzyme (figure 18.11).
DNA techniques have been used to produce humanized monoclonal Western Blotting
antibodies. In the Western blotting technique, the various proteins that make up
Quantifying Antigen-Antibody Reactions an antigen are separated by size before reacting them with antibodies
(figure 18.13).
The concentration of antibody molecules in a specimen is usually deter-
mined by making serial dilutions; the last dilution that gives a detectable Fluorescence-Activated Cell Sorter (FACS)
antigen-antibody reaction reflects the titer. The fluorescence-activated cell sorter (FACS) can be used to count
and separate antigens labeled with fluorescent antibodies.
18.4 ■ Observing Antigen-Antibody Aggregates
Precipitation Reactions
Antibodies bound to soluble antigens may form complexes that pre-
cipitate out of solution (figure 18.5). An example of a test that involves
a precipitation reaction is the Ouchterlony technique (figures 18.6).

Review Questions
Short Answer Multiple Choice
1. How is immune globulin different from hyperimmune globulin? 1. Which is an example of immunization that elicits active immunity?
2. Describe two advantages of an attenuated vaccine over an inacti- a) Giving antibodies against diphtheria
vated one. b) Immune globulin injections to prevent hepatitis
3. Describe two advantages of an inactivated vaccine over an attenu- c) Sabin polio immunization
ated one. d) Rabies immune globulin
4. What is herd immunity? e) Tetanus immune globulin
5. Describe how both active and passive immunization can be used to 2. Breast feeding provides which of the following to an infant?
combat tetanus. a) Artificial active immunity
6. Why are humanized monoclonal antibodies better for therapy than b) Artificial passive immunity
the original versions? c) Natural active immunity
7. In a precipitation reaction, what is meant by “optimal propor- d) Natural passive immunity
tions”? 3. Vaccines ideally should be all of the following, except
8. Is blood typing an example of a precipitation reaction or an agglu- a) effective in protecting against the disease.
tination reaction?
b) inexpensive.
9. An ELISA test is used to screen patient specimens for HIV. A posi- c) stable.
tive ELISA test is confirmed by a Western blot test. Why not the
d) living.
other way around, with the ELISA second?
e) easily administered.
10. What is the purpose of anti-human IgG antibodies in immunologi-
cal testing?
436 Chapter 18 Applications of Immune Responses

4. Severely immunosuppressed people should not receive the measles 9. All of the following are matching pairs except
vaccine. Based on this information, the vaccine is a) ELISA—radioactive label.
a) an inactivated whole agent. b) fluorescence-activated cell sorter—flow cytometry.
b) a toxoid. c) fluorescent antibody test—microscopy.
c) a subunit vaccine. d) Western blot—gel electrophoresis.
d) a genetically engineered vaccine against hepatitis B. 10. Which of the following would be most useful for screening thou-
e) an attenuated vaccine. sands of specimens for antibodies that indicate a certain disease?
5. All of the following are attenuated vaccines except a) Western blot b) Fluorescent antibody c) ELISA
a) chickenpox. b) mumps. c) rubella. d) All of the above e) None of the above
d) Salk polio. e) yellow fever.
6. An important subunit vaccine that is widely used is the Applications
a) pertussis vaccine. b) Sabin vaccine. c) Salk vaccine. 1. A new parent asks you which vaccines the CDC recommends for a
2-month-old infant. What is your answer? The chapter 18 readings
d) measles vaccine. e) mumps vaccine.
at the text website (www.mhhe.com/nester7) provide a link to
7. In quantifying antibodies in a patient’s serum the CDC’s recommended immunization schedules.
a) total protein in the serum is measured. 2. There has been debate about keeping smallpox virus stored, since
b) the antibody is usually measured in grams per ml. the disease has been eradicated. What would be an argument for
c) the serum is serially diluted. keeping the virus? What should be done to protect against use of
d) both antigen and antibody are diluted. the virus in biological warfare?
e) the titer refers to the amount of antigen added.
8. Which of the following about immunological testing is false? Critical Thinking +
a) Polyclonal antibody preparations recognize multiple epitopes. 1. In figure 18.5, how would the curve change if the concentration of
b) Monoclonal antibodies recognize a single epitope. antibody in the original sample were increased? (Would the shape
c) Serum and plasma can both be tested for antibodies. of the curve change? Would the curve be shifted left, right, up, or
down?) Briefly explain your answer.
d) The direct ELISA uses anti-human IgG antibodies.
e) A rise in specific antibody titer indicates an active infection. 2. Staphylococcus aureus makes a protein called protein A, which
binds to the Fc region of antibody molecules from a wide variety
of species. How could protein A be exploited in immunoassays?
19 Epidemiology
KEY TERMS
Attack Rate The percentage of
individuals developing illness in a
population exposed to an infectious
agent.
Incidence The number of new
cases of a disease in a population at
risk during a specified time period.
Morbidity Illness; most often
Case-Fatality Rate Percentage expressed as the rate of illness in a
of people dying of a given disease given population at risk.
within a specific time period.
Mortality Death; most often
Communicable Disease An expressed as a rate of death in a
infectious disease that can be given population at risk.
transmitted from one host to another.
Outbreak A cluster of cases
Endemic A disease or other occurring during a brief time interval
occurrence that is constantly present and affecting a specific population.
in a population.
Pandemic A worldwide epidemic.
Epidemic A disease or other
Prevalence The total number
occurrence that has a much higher
of cases of a disease in a given
incidence than usual.
population at any time or for a
Herd Immunity A phenomenon specific period.
that occurs when a critical
Reservoir of Infection The natural
concentration of immune hosts
habitat of a pathogen; sum of the
prevents the spread of an infectious
potential sources of an infectious
agent, thereby protecting the entire
agent.
population.
CDC microbiologist studies influenza virus.

A Glimpse of History
Puerperal fever, a bacterial infection of the uterus following childbirth,
strong disinfectant before attending patients. The incidence of puerperal
rose to epidemic proportions in the eighteenth century when more women
fever dropped to one-third its previous level. Instead of appreciating these
chose to deliver their babies in hospitals. By the middle of the nineteenth
findings, Semmelweis’s colleagues refused to admit responsibility for
century in the hospitals of Vienna, the major medical center of the world
the deaths of so many patients. His work was so fiercely attacked that he
at that time, about one of every eight women died of puerperal fever fol-
was forced to leave Vienna and return to his native Hungary. Although
lowing childbirth.
his disinfection techniques achieved a remarkable reduction in the num-
In 1841, Ignaz Semmelweis, a Hungarian, traveled to Vienna to
ber of deaths from puerperal fever there as well, Semmelweis became
study medicine. After medical school, he worked for Professor Johann
increasingly outspoken and bitter. He was finally confined to a mental
Klein, who managed one section of the Lying-In Hospital with the medi-
institution, where he died one month later of a generalized infection simi-
cal students. Midwives and midwifery students served a second section.
lar to the kind that had killed his friend and the many women who had
Semmelweis noticed that the incidence of puerperal fever in Dr. Klein’s
contracted puerperal fever following childbirth.
section was as high as 18%, four times that in the section served by the
midwives. During this period, a friend of Semmelweis incurred a scalpel

E 
wound while doing an autopsy and died of symptoms very similar to pidemiology is the study of disease patterns in populations.
those of puerperal fever. Semmelweiss reasoned that the “poison” that Epidemiologists—the “disease detectives”—collect and
killed his friend probably also contaminated the hands of the medical compile data about sources of disease and associated risk
students who did autopsies. Perhaps these students were transferring the
factors to design infection control strategies and to prevent or
“poison” from the cadavers to the women in childbirth. Midwives, after
all, did not perform autopsies. This was before Pasteur and Koch estab-
predict the spread of disease. They approach a disease outbreak
lished the germ theory of disease, so Semmelweis had no way of knowing much as a criminal detective describes the scene of a crime using
that the “poison” being transferred was probably Streptococcus pyogenes, expertise in diverse disciplines including ecology, microbiology,
a common cause of many infections, including puerperal fever. sociology, statistics, and psychology. Many of our daily habits,
To test his hypothesis that physicians and students were transfer- from handwashing to waste disposal, are based on the work of
ring “poison” to patients, Semmelweis had them wash their hands with a epidemiologists.

437
438 Chapter 19 Epidemiology

19.1 ■ Principles of Epidemiology often associated with non-communicable diseases such as cancer
or heart attack. In developing countries, however, communicable
Learning Outcomes diseases are still a major cause of death. Case-fatality rate is
1. Explain why epidemiologists are most concerned with the rate of
the percentage of a population that dies from a specific disease.
disease rather than the number of cases. Diseases such as plague and Ebola hemorrhagic fever are feared
2. Describe movement of a pathogen from its reservoir to a host, and
because of their very high case-fatality rate. As the case-fatality
back to its reservoir or to another host. rate for AIDS has decreased due to improved treatment, the preva-
3. Explain how characteristics of a pathogen or of a host can
lence of AIDS has increased because more people with the disease
influence the epidemiology of a disease. survive within the population.
Endemic diseases are constantly present in a given popula-
Diseases that can be transmitted from one host to another, such as tion. For example, the common cold is endemic in the United
measles, colds, and influenza, are contagious, or communicable, States. When cases occur only from time to time, they are
diseases. Disease transmission is determined by interactions sporadic. An unusually large number of cases in a population
between the environment, the pathogen, and the host. Control of constitutes an epidemic. Epidemics may be caused by diseases
any of these factors may break the cycle of infection, hindering or not normally present in a population, such as cholera being rein-
preventing the disease. For example, improved sanitation prevents troduced to the Western Hemisphere, or by fluctuations in the
the pathogen from accessing the host; antimicrobial medications incidence of endemic diseases such as influenza and pneumonia
can kill or inhibit infecting organisms; and vaccination increases (figure 19.1). An outbreak describes a cluster of cases occurring
resistance of the host. during a brief time interval and affecting a specific population; an
Diseases that do not spread from one host to another are outbreak may herald the onset of an epidemic. When an epidemic
called non-communicable diseases. Microorganisms that cause spreads worldwide, as AIDS has, it is called a pandemic.
these diseases most often arise from an individual’s normal micro-
biota or from the environment. Clostridium tetani, the bacterium Reservoirs of Infection
that causes tetanus, can enter a host from the environment but is
not then transmitted to another host. A pathogen must have a suitable environment in which to live.
That natural habitat, the reservoir of infection, may be on or in
an animal, including humans, or in an environment such as soil or
Rate of Disease in a Population water (figure 19.2). The reservoir of infection affects the extent
Epidemiologists generally are less concerned with the absolute
number of cases of a disease than they are with the rate. For
10
example, 100 people in a city of 1,000,000 developing genital
herpes in a given time period is not as alarming as 100 people
in a town of 5,000 developing the same disease over the same
time period. The much higher rate or proportion of the population
infected is of greater concern to the epidemiologist. 8

The attack rate describes the percentage of people who

become ill in a population after exposure to an infectious agent.
Percent of deaths

For example, if 100 people at a party eat chicken contaminated

with Salmonella, and 10 people develop symptoms of salmonel- 6
losis, then the attack rate is 10%. The attack rate reflects many fac-
tors, including the infectious dose of the organism and the immune
status of the population.
The incidence of a disease is the number of new cases in a 4 Epidemic threshold
specific time period in a given population; the prevalence of a Seasonal (endemic) baseline
Actual percentage of total
disease is the total number of cases at any time or for a specific deaths in the population
period in a given population. Usually these are expressed as a
rate, the number of cases per 100,000 people. Incidence provides
0
a useful measure of the risk of an individual contracting the dis-
5 15 25 35 45 5 15 25 35 45 5 15 25 35 45 5 15 25 35 45 5
ease. Prevalence reflects the overall impact of a disease on society
Year 1 Year 2 Year 3 Year 4
because it includes both old and new cases, taking the duration of
Weeks of the year
the disease into account.
Morbidity refers to the incidence of disease in a population FIGURE 19.1 Endemic Disease Can Be Epidemic Example of
at risk. Contagious diseases such as influenza often have a high yearly fluctuation of pneumonia and influenza deaths (expressed as a
morbidity rate because each infected individual may transmit the percentage of all deaths).
infection to several others. Mortality refers to the overall death ? During what season(s) of the year is a pneumonia epidemic most
rate in a population. In developed countries, mortality is most likely?
 Part III Microorganisms and Humans 439

may shed the organism intermittently or constantly for months,

years, or even a lifetime.
Some carriers have an asymptomatic infection; their immune
system is actively responding to the invading microorganism, but
they have no obvious clinical symptoms. Because these people
Humans often have no reason to consider themselves a reservoir, they
move freely about, spreading the pathogen. People with asymp-
tomatic infections are a significant complicating factor in the
control of sexually transmitted infections (STIs) such as gonor-
rhea. Up to 50% of women infected with Neisseria gonorrhoeae
Animals have no symptoms, and may unknowingly transmit the disease
(domestic) to their sexual partners. In contrast, infected men are more often
symptomatic and seek medical treatment. Gonorrhea infections
Environment
can be treated with antibacterial drugs, but locating sexual con-
tacts of infected people is often difficult and costly. Neisseria
gonorrhoeae, p. 622
Some potentially pathogenic microbes can colonize the skin
Animals or mucosal surfaces, establishing themselves as part of a person’s
(wild) microbiota. For instance, many people carry Staphylococcus
aureus as a part of their nasal or skin microbiota. Carriers of
FIGURE 19.2 Reservoirs of Infection
S. aureus may never have any illness or disease as a result of
? How might you be a reservoir of infection? the organism, but they remain a potential source of infection to
themselves and others. Unfortunately, ridding a colonized carrier
of the infectious organism is often difficult, even with the use of
and distribution of a disease. Once the reservoir is identified, peo- antimicrobial drugs. Staphylococcus aureus, p. 524
ple can be prevented from coming into contact with the disease
source. The United States does not have epidemics of plague, in Non-Human Animal Reservoirs
part because populations of wild rats, mice, and prairie dogs are Non-human animal reservoirs are the source of many pathogens.
controlled. These are the natural reservoirs of Yersinia pestis, the Poultry are a reservoir of gastrointestinal pathogens such as spe-
bacterium that causes plague. Yersinia pestis, p. 678 cies of Campylobacter and Salmonella. In the United States,
raccoons, skunks, and bats are reservoirs of the rabies virus.
Human Reservoirs Occasional transmission of plague to humans is still reported in
Infected humans are a significant reservoir of most communica- the southwestern states where Y. pestis is endemic in prairie dogs
ble diseases. In some cases, humans are the only reservoir. In and other rodents. Rodents, particularly the deer mouse, are the
other instances, the pathogen can also exist in other animals and, reservoir for hantavirus. Campylobacter, p. 593  rabies, p. 658
occasionally, in the environment as well. When infected humans Diseases such as plague and rabies that can be transmitted to
are the only reservoir, the disease can be easier to control because humans but primarily exist in other animals are called zoonotic
it is more feasible to set up prevention and control programs diseases, or zoonoses. Zoonotic diseases are often more severe
in humans than in wild animals. The eradication of smallpox is in humans than in the typical animal host because the infection
an excellent example. The combined effects of widespread vac- in humans is accidental; there has been no evolution toward the
cination programs (which resulted in fewer susceptible people) balanced pathogenicity that normally exists between host and
and isolation of those who became infected eliminated the small- parasite. balanced pathogenicity, p. 385
pox virus from nature. The virus no longer had a reservoir in
which to multiply. Environmental Reservoirs
Some pathogens have environmental reservoirs. Clostridium
Symptomatic Infections People with symptomatic illnesses are
botulinum, which causes botulism, and Clostridium tetani, which
an obvious source of infectious agents. Ideally they understand the
causes tetanus, are both widespread in soils. Pathogens that have
importance of taking precautions to avoid transmitting their illness
environmental reservoirs are difficult or impossible to eliminate.
to others. Staying home and resting while ill both helps the body
Clostridium botulinum, p. 652 Clostridium tetani, p. 555
recover and protects others from exposure to the disease-causing
agent. Even conscientious people, however, can unintentionally
be a source of infection. For example, people who are in the Portals of Exit and Portals of Entry
incubation period of mumps shed virus before symptoms appear.
incubation period, p. 383 mumps, p. 583 A pathogen must leave its reservoir to be transmitted to a suscep-
tible host. If the reservoir is an animal, the body orifice or surface
Asymptomatic Carriers A person can harbor a pathogen with no from which a microbe is shed is called the portal of exit. To com-
ill effects, acting as a carrier of the disease agent. These people plete the cycle of infection, the pathogen is transmitted to the next
440 Chapter 19 Epidemiology

Disease Transmission
Transmission of a pathogen from one person to another through
Eyes Ears
the air, by physical contact, by ingestion of food or water, or via
a living agent such as an insect is called horizontal transmission
Respiratory tract
(figure 19.4). This contrasts with vertical transmission, which
Digestive is the transfer of a pathogen from a pregnant woman to her fetus,
tract or from a mother to her infant during childbirth or breast feeding.

Contact
Transmission of a pathogen from one host to another often
involves direct or indirect contact.

Direct Contact Direct contact can be as simple as a handshake,

Broken skin or as intimate as sexual intercourse. Sometimes the transfer of very
few microbes can initiate an infection. For example, the infectious
dose of Shigella species, which are intestinal pathogens, is about
Genitourinary tract 10 to 100 cells, a number easily transferred when shaking hands.
Once on the hands, the organisms can inadvertently be ingested—
resulting in fecal-oral transmission. Handwashing, a simple rou-
tine that physically removes microbes, is important in preventing
FIGURE 19.3
this type of spread of disease. Even washing in plain water
Portals of Entry
reduces the numbers of potential pathogens on the hands, which
? How does lysozyme decreases the possibility of transferring or ingesting enough cells
in the mucus that
to establish an infection. In fact, routine handwashing is consid-
lines the portals of
entry protect against ered to be the single most important measure for preventing the
infection? spread of infectious disease. When handwashing is not possible,
use of alcohol gels may provide some protection, but they are not
effective against all pathogens. Shigella sp., p. 588
Pathogens that cannot survive for extended periods in the
host and enters through a body surface or orifice called the portal
environment must generally be transmitted through direct con-
of entry (figure 19.3).
tact. Treponema pallidum, which causes syphilis, and Neisseria
Microorganisms that inhabit the intestinal tract are shed in the
gonorrhoeae, which causes gonorrhea, both die quickly when
feces. Pathogens such as Vibrio cholerae, which cause massive
exposed to a relatively cold, dry environment and thus require
volumes of watery diarrhea, can contaminate drinking water and
intimate sexual contact for their transmission. Treponema pallidum,
food. Pathogens such as Mycobacterium tuberculosis and various
p. 627 Neisseria gonorrhoeae, p. 622
respiratory viruses exit the body in droplets of saliva and mucus
when people talk, laugh, sing, sneeze, or cough. Organisms that Indirect Contact Indirect contact involves transfer of pathogens
inhabit the skin are constantly shed on skin cells. Even as you read via inanimate objects, or fomites, such as clothing, table-tops,
this text you are shedding skin cells, some of which may have doorknobs, and drinking glasses. As an example, carriers of
Staphylococcus aureus on their surface. Genital pathogens such Staphylococcus aureus may inoculate their fingers with the organ-
as Neisseria gonorrhoeae can be carried in semen and vaginal ism when touching a skin lesion or colonized nostril. Organisms
secretions. Hantavirus is found in the saliva, urine, and droppings on the fingers can then easily be transferred to a fomite. Another
of deer mice. Vibrio cholerae, p.  586 Mycobacterium tuberculosis, person can acquire the microbes when handling that object. Again,
p. 502 hantavirus, p. 512 handwashing is an important control measure.
To cause disease, not only must a pathogen be transmitted
to a new host, it must also colonize a surface of or enter that Droplet Transmission Large microbe-laden respiratory droplets
new host. Cells of a Shigella species can be transferred by a generally fall to the ground no farther than a meter (approximately
handshake, for example, but they will only cause disease if they 3 feet) from release. People in close proximity can inhale those
are then transferred to the mouth or to food that is then eaten. In droplets, resulting in the spread of respiratory disease via droplet
this case, the mouth is the portal of entry. Respiratory pathogens transmission. Although physical contact is not necessary, droplet
released into the air during a cough generally cause disease only transmission is considered contact transmission because of the
when someone inhales them. For these pathogens, the nose is close range involved. Droplet transmission is particularly impor-
the portal of entry. Many organisms can cause disease if they tant as a source of contamination in densely populated buildings
enter one body site, but are harmless if they enter another. For such as schools and military barracks. Desks or beds in such loca-
instance, Enterococcus faecalis may cause a bladder infection if tions should be spaced more than 1.5 meters and preferably about
it enters the urinary tract, but it is harmless when it is ingested 3 meters apart to minimize the transfer of infectious agents. The
and then colonizes the large intestine, where it may reside as part spread of respiratory diseases is minimized if people cover their
of the normal microbiota. mouths when they cough or sneeze.
 Part III Microorganisms and Humans 441

Airborne

Indirect contact
Vectors

Direct contact Food/water

FIGURE 19.4 Transmission of Pathogens Microbes may be spread by indirect contact with fomites, by direct contact, through the air,
by vectors, or by food or water.

? List three potential fomites that you contacted today.

Food and Water As mentioned earlier, when people talk, laugh, sing, sneeze,
Pathogens, particularly those that infect the digestive tract, can be or cough they continually discharge microorganisms in liquid
transmitted through contaminated food or water. Foods can become droplets. Large droplets quickly fall to the ground, but smaller
contaminated in a number of ways. Animal products such as meat droplets dry, creating droplet nuclei composed of microbes
and eggs can have pathogens that originated from the animal’s intes- attached to a thin coat of the dried material. These can remain
tinal tract. This is the case with poultry contaminated with species suspended indefinitely in the presence of even slight air currents.
of Salmonella or Campylobacter and hamburger contaminated with Other airborne particles, including dead skin cells, household dust,
E. coli O157:H7. Pathogens can also be inadvertently added dur- and soil disturbed by the wind, may also carry respiratory patho-
ing food preparation. Staphylococcus aureus carriers who do not gens. An air-conditioning system can distribute contaminated air.
wash their hands prior to preparing food can easily contaminate the The number of viable organisms in air can be estimated by
food. Cross-contamination results when pathogens from one food using a machine that pumps a measured volume of air, including
are transferred to another. A cutting board used first to carve raw any suspended dust and particles, against the surface of a nutrient-
chicken and then to cut cooked potatoes can transfer Salmonella cells rich medium in a Petri dish. This technique has shown that the
from the chicken onto the potatoes. Because many foods are a rich number of bacteria in the sampled air rises in proportion to the
nutrient source, microorganisms can multiply to high numbers if the number of people in a room (figure 19.5).
contaminated food is not refrigerated. Sound food-handling methods,
including sanitary preparation as well as thorough cooking and proper
storage, can prevent foodborne diseases. foodborne disease, p. 756
Waterborne disease outbreaks can involve large numbers of
people because municipal water systems distribute water to wide-
spread areas. The 1993 waterborne outbreak of Cryptosporidium
parvum, an intestinal parasite, in Milwaukee, Wisconsin, was esti-
mated to have involved over 400,000 people. Prevention of water-
borne diseases requires disinfection and filtration of drinking water
and proper disposal and treatment of sewage. Cryptosporidium
parvum, p. 604 drinking water treatment, p. 742  sewage treatment, p. 736

Air (a) (b)

Respiratory diseases are often transmitted through the air. When FIGURE 19.5 Air Sample Cultures (a) Air from a clean, empty
particles larger than 10 μm are inhaled, they are usually trapped in hospital room. (b) Air from a small room containing 12 people. In
the mucus lining of the nose and throat and eventually swallowed. both situations, 5 cubic feet of air was sampled.
Smaller particles, however, can enter the lungs, where any patho- ? Name an organism that you might expect to find in the plate
gens they carry can potentially cause disease. shown in (b).
442 Chapter 19 Epidemiology

Understandably, airborne transmission of pathogens is very Prevention of vector-borne disease relies largely on control
difficult to control. To prevent the buildup of airborne pathogens, of arthropods. Malaria, once endemic in the continental United
modern public buildings have ventilation systems that constantly States, was successfully eliminated from the nation through a
change the air. The air pressure in hospital microbiology laborato- combination of mosquito elimination and prompt treatment of
ries can be lowered so that air flows in from the corridors, prevent- infected patients. Unfortunately, worldwide eradication efforts
ing microorganisms and viruses from being swept out of the lab to that initially showed great promise ultimately failed, in part due to
other parts of the building. Air in specialized hospital rooms, jet- the decreased vigilance that accompanied the dramatic but short-
liners, and some laboratories is circulated through high-efficiency lived decline of the disease.
particulate air (HEPA) filters to remove airborne organisms that
may be present. HEPA filters, p. 115 MicroByte
An outbreak of dengue fever in Key West, Florida, in 2009 was
transmitted by Aedes mosquitoes that bite during the day.
Vectors
A vector is any living organism that can carry a disease-causing
microbe, but most commonly they are arthropods such as mos- Pathogen Factors That Influence
quitoes, flies, fleas, lice, and ticks. A vector can carry a pathogen
externally or internally.
the Epidemiology of Disease
Flies that land on feces can pick up intestinal pathogens such An infectious agent transmitted to a new host can potentially cause
as Escherichia coli O157:H7 and Shigella species on their legs. disease in that host. Some pathogens produce more disease than
If the fly then lands on food and transfers the microorganisms, it others. For example, 95% of people infected with the measles virus
serves as a mechanical vector, carrying the microbe on its body will develop symptoms of measles. On the other hand, less than
from one place to another (figure 19.6a). 1% of those infected with poliovirus will develop poliomyelitis.
Diseases such as malaria, plague, and Lyme disease are The outcome of transmission is affected by many factors, including
transmitted via arthropods that harbor the pathogen internally. virulence of the pathogen, the dose, and the incubation period.
The vector either injects the infectious agent while taking a blood
meal, or deposits the pathogen when it defecates on skin where Virulence
it can be inoculated when the individual scratches the bite. For Successful pathogens have virulence factors that increase their
example, infected fleas inject Yersinia pestis while attempting to ability to cause disease, as discussed in chapter 16. These may
take a blood meal. In the case of malaria, caused by species of the allow pathogens to adhere to a host or penetrate host cells.
eukaryotic pathogen Plasmodium, the mosquito not only transmits Neisseria gonorrhoeae binds to mucosal epithelial cells; Shigella
the parasite but also plays an essential role in its reproductive life species adhere to intestinal epithelial cells and promote their own
cycle. Such vectors are called biological vectors (figure 19.6b). uptake. Other virulence factors allow pathogens to thwart immune
The pathogen multiplies to high numbers within this vector. defenses of the host. Capsules such as those of Streptococcus
 Plasmodium, p. 292 pneumoniae often protect microbial cells from phagocytosis.

FIGURE 19.6 Vector

Transmission (a) A
mechanical vector moves
microbes from one place
to another. (b) A biologi-
cal vector participates
in the life cycle of the
pathogen and provides a
place for it to multiply.

? Name one disease

spread by a
mechanical vector (a) Mechanical vector
and one disease
involving a biological
vector.

(b) Biological vector

 Part III Microorganisms and Humans 443

Mycobacterium tuberculosis can survive within activated macro- agent cannot spread in a population because most potential hosts
phages. Finally, pathogens damage the host, perhaps by produc- are immune. Unfortunately, some infectious agents can undergo
tion of toxins or destructive enzymes, or perhaps by stimulating antigenic variation and overcome herd immunity. Currently,
a strong inflammatory response that damages host tissue. E. coli humans have no herd immunity to the H5N1 (avian) influenza
O157:H7 produces a cytotoxin that destroys cells that line blood strain. antigenic variation, pp. 178, 390
vessels; Clostridium perfringens, the cause of gas gangrene,
destroys host cell membranes with a phospholipase. Organisms General Health
that possess a variety of virulence factors are more likely to Malnutrition, overcrowding, and fatigue increase people’s sus-
cause severe disease. virulence factors, p. 383 S. pneumoniae, p. 497 ceptibility to infectious disease, enhancing its spread. Infectious
C. perfringens, p. 558 diseases are more of a problem in developing areas of the world
where individuals are crowded together without proper food or
The Dose sanitation. Factors that promote good general health result in
The probability of infection and disease is generally lower when increased resistance to diseases such as tuberculosis. When infec-
an individual is exposed to small numbers of a pathogen. This is tion does occur in a healthy individual, it is more likely to be
because a certain minimum number of cells of the pathogen must asymptomatic or result in mild disease.
enter the body and produce enough damage to cause disease. If 30
cells enter the body, yet 1,000,000 cells are required to produce Age
symptoms, then it will take some time for the bacterial population The very young and the elderly are generally more susceptible
to increase to that number. Because host defenses are mobilized to infectious agents. The immune system of young children is
at the same time and are racing to eliminate the invader, small not fully developed and, consequently, they are vulnerable to
doses often result in asymptomatic infections—the immune sys- certain diseases. For example, young children are particularly
tem eliminates the organism before symptoms appear. On the susceptible to meningitis caused by Haemophilus influenzae.
other hand, there are few if any infections for which immunity is The introduction of Hib vaccine in the late 1980s has dramati-
absolute. An unusually large exposure to a pathogen, such as can cally decreased the incidence of meningitis in this age group. The
occur in a laboratory accident, may produce serious disease in a elderly are more prone to disease because immunity wanes over
person who is ordinarily immune to that microbe. Therefore, even time. Influenza outbreaks in nursing homes often have high case-
immunized persons should take precautions to minimize exposure fatality rates. Older adults are also less likely to update their
to infectious agents. This principle is especially important for immunizations, making them more susceptible to diseases such
medical workers taking care of patients with infectious diseases. as tetanus (figure 19.7). To prevent tetanus, a booster vaccine is
needed once every 10 years. Hib vaccine, p. 647 influenza, p. 508
The Incubation Period tetanus, p. 555
The extent of the spread of an infectious agent is influenced by
its incubation period. Diseases with typically long incubation
periods can spread extensively before the first symptomatic cases 20
appear. An excellent example was the spread of typhoid fever
from a ski resort in Switzerland in 1963. As many as 10,000 peo- 18
ple were exposed to drinking water containing small numbers of
Salmonella enterica serotype Typhi, the bacterium that causes the 16
disease. The long incubation period of the disease, 10 to 14 days, 14
allowed the organism to be spread by the skiers, who flew home to
Number of cases

various parts of the world before they became ill. As a result, more 12
than 430 cases of typhoid fever appeared in at least six countries.
typhoid fever, p. 592 incubation period, p. 383 10

8
Host Factors That Influence
6
the Epidemiology of Disease
Some populations more than others are likely to be affected by 4
a given disease-causing agent. Many population characteristics
2
influence the occurrence of disease.
0
Immunity to the Pathogen 0–4 5–14 15–24 25–39 40–64 >65
Previous exposure or immunization of a population to a disease Age
agent or an antigenically related agent decreases incidence of the
disease. A disease is unlikely to spread in a population in which FIGURE 19.7 New Cases of Tetanus by Age Group
90% of the individuals are immune to the disease agent. Herd ? Why do you think individuals from 40–64 contract tetanus more
immunity protects non-immune individuals when an infectious often than younger people?
444 Chapter 19 Epidemiology

Gender the relationship between microbes and disease was accepted, he

Gender may influence disease distribution. Women are more documented that the cholera epidemics plaguing England from
likely to develop urinary tract infections because their urethra (the 1849 to 1854 were due to contaminated water supplies. He did
tube that connects the urinary bladder to the external environment) this by carefully comparing the conditions of households that were
is relatively short. Microbes can ascend the urethra into the blad- affected by cholera to those that were not, eventually determining
der. Pregnant women are more susceptible to listeriosis, caused by that the primary difference was their water supply. His analysis
Listeria monocytogenes. listeriosis, p. 648 saved countless lives. cholera, p. 586

Religious and Cultural Practices Descriptive Studies

The distribution of disease is influenced by religious and cultural
When a disease outbreak occurs, epidemiologists conduct a
practices. An infant who is breast fed is less likely to have infec-
descriptive study by collecting data that characterize the occur-
tious diarrhea because of the protective antibodies in the mother’s
rence, from the time and place of the outbreak to the individuals
milk. Groups who eat traditional dishes made from raw freshwater
affected. That information is used to compile a list of possible risk
fish are more likely to acquire the tapeworm, Diphyllobothrium
factors involved in the spread of disease.
latum, a parasite normally killed by cooking.

Genetic Background The Person

Determining the profile of those who become ill is critical to
Natural immunity can vary with genetic background, but it is usually
defining the population at risk. Variables such as age, gender,
difficult to determine the relative importance of genetic, cultural,
ethnicity, occupation, personal habits, previous illnesses, socio-
and environmental factors. In a few instances, however, the genetic
economic class, and marital status may all yield clues about risk
basis for resistance to infectious disease is known. For example,
factors for developing the disease. For example, in the Swiss ski
many people of black African ancestry are not susceptible to malaria
resort epidemic of typhoid fever mentioned earlier, cases occurred
caused by Plasmodium vivax because they lack a specific red blood
only in tourists because the local people rarely drank water, pre-
cell receptor used by the infecting organism. Some populations of
ferring wine instead.
Northern European ancestry are less susceptible to HIV infection
because they lack a certain receptor on their white blood cells.
The Place
MicroAssessment 19.1 The geographic location of disease acquisition identifies the gen-
eral site of contact between the person and the infectious agent
Rates of disease within a population are a concern of epidemiologists (see figure 21.28). This helps pinpoint the exact source. The loca-
who study disease patterns. Disease may be prevented by controlling
tion may also give clues about potential reservoirs, vectors, or
the environment, the pathogen, or the host. The reservoir of a disease
agent can be infected people, other animals, or the environment. To geographical boundaries that might affect disease transmission.
spread, infectious microbes must exit one host and enter another. For example, malaria can be transmitted only in regions that have
Handwashing and vector control can prevent many diseases; airborne the appropriate mosquito vector.
transmission of pathogens is difficult to control. The outcome of
transmission is affected by the virulence and dose of the infecting MicroByte
agent, the incubation period, and by various characteristics of the The first case of H1N1 influenza in the pandemic of 2009 was traced
host population. to a 5-year-old boy in Veracruz, Mexico.
1. Explain the difference between incidence of a disease and
prevalence of the disease.
The Time
2. Explain how a low dose of an infecting organism can result in
an asymptomatic infection.
The timing of an outbreak may also yield clues. A rapid rise in the
numbers of people who become ill suggests that they were all exposed
3. Considering that circulating blood is not normally released
from the body, describe how blood-borne microbes might exit. + to a single, common source of the infectious agent, such as contami-
nated chicken at a picnic. This is called a common-source epidemic.
In contrast, if the numbers of ill people rise gradually, then the dis-
ease is likely contagious, with one person transmitting it to several
19.2 ■ Epidemiological Studies others, who each then transmit it to several more, and so on. This is
called a propagated epidemic (figure 19.8). The first case in such
Learning Outcomes an outbreak is called the index case. The time between the onset
4. Compare and contrast descriptive studies, analytical studies, and of symptoms in this case and the next cases reflects the incubation
experimental studies. period of the disease.
5. Describe how a common-source epidemic can be distinguished The season in which the epidemic occurs may also be sig-
from a propagated epidemic. nificant. Respiratory diseases including influenza, respiratory
syncytial virus infections, and the common cold are more eas-
British physician John Snow illustrated the power of a well- ily transmitted in crowded indoor conditions during the win-
designed epidemiological study over 150 years ago. Long before ter. Conversely, vector and foodborne diseases are more often
 Part III Microorganisms and Humans 445

Cross-Sectional Studies
A cross-sectional study surveys a range of people to determine
the prevalence of such characteristics as infection, presence of risk
Common-source outbreak
Propagated epidemic
factors associated with disease, or previous exposure to a disease-
causing agent. This population assessment may suggest associa-
Number of new cases

tions between risk factors and disease. A cross-sectional survey

does not attempt to establish cause of disease.

Retrospective Studies
A retrospective study is done following a disease outbreak.
The actions and events surrounding clinical cases (individuals
who developed the disease) are compared to those surrounding
appropriate controls (those who remained healthy). A case-
control study starts with the disease and attempts to identify
the causative chain of events leading to it. If an activity or
1 2 3 4 5 6 7 event is common among the cases, but not among the controls,
Days
it may have been a factor in the development of the disease.
FIGURE 19.8 Comparison of Common-Source Versus It is important to select controls that match the cases with
Propagated Epidemics The graph depicts the number of new cases respect to variables not thought to be associated with disease.
that develop over a period of days. Matching variables such as age, gender, and socioeconomic
? How does comparing day 1 and day 7 help you to determine if an status, ensures that all controls had equal probability of com-
epidemic is common-source or propagated? ing in contact with the disease agent.

transmitted in warm weather when people are more likely to be Prospective Studies
exposed to mosquitoes and ticks, or eat picnic food that has not A prospective study looks ahead to see if the risk factors iden-
been stored properly (figure 19.9). tified by the retrospective study predict a tendency to develop
the disease. Cohort groups, which are study groups that have a
known exposure to the risk factor, are selected and then followed
Analytical Studies over time. The incidence of disease in those who were exposed
Analytical studies are designed to determine which of the poten- to the risk factor and those who were not is then compared. By
tial risk factors identified by the descriptive studies are actually following cohort groups, the study attempts to determine if the
relevant in the spread of the disease. suspected cause does indeed correlate with the expected effect.
This type of study is less prone to the bias of inaccurate recall than
a retrospective study. It is generally more time-consuming and
expensive, however, particularly when examining a disease with a
Respiratory syncytial virus
Campylobacter long incubation period.

Experimental Studies
Presence of disease

An experimental study is used to judge the cause-and-effect

relationship of the risk factors or, more commonly, the pre-
ventive factors, and the development of disease. Experimental
studies are done most frequently to assess the value of a par-
ticular intervention or treatment, such as antimicrobial drug
therapy. The effectiveness of the treatment is compared with
one of known value or with a placebo. A placebo is a mock
drug—it looks and tastes like the experimental drug but has
no medicinal value. To assess the value of the experimental
Jan. June Jan. June Jan. June Jan. drug, a group of patients is divided into two subgroups, one
Year 1 Year 2 Year 3 of which is given the treatment and the other an alternative or
Month a placebo. To avoid bias, the study should ideally be double-
blind, where neither the researchers nor the patients know
FIGURE 19.9 Seasonal Occurrence of Respiratory who is receiving the experimental treatment. Ethical issues
and Gastrointestinal Infections sometimes necessitate the use of animals rather than human
? Why does the level of foodborne disease increase in the summer? patients in experimental studies.
446 Chapter 19 Epidemiology

MicroAssessment 19.2 both recognizing and reporting disease cases to public health
authorities. This information can alert officials to changes in inci-
Descriptive epidemiological studies attempt to identify the potential dence or prevalence of a disease or it may provide clues as to the
risk factors that lead to disease. Analytical studies try to determine
which factors are actually relevant to disease development.
cause of disease. In some cases, cooperative surveillance efforts
Experimental studies are generally used to evaluate the effectiveness coupled with global immunization programs and isolation of cases
of a treatment or an intervention in preventing disease. can result in eradication of disease, as was the case with smallpox.
4. On what three factors does a descriptive study focus?
5. How is the timing of a propagated epidemic related to the National Disease Surveillance Network
incubation period of the pathogen?
Infectious disease control depends on a network of agencies
6. Why is it important to include a placebo in a scientific study to across the country to monitor disease development. It is partly
assess the effectiveness of a drug? +
because of this network that infectious diseases do not claim more
lives in the United States.

19.3 ■ Infectious Disease Centers for Disease Control and Prevention

Surveillance The Centers for Disease Control and Prevention (CDC) in
Atlanta, Georgia, is part of the U.S. Department of Health and
Learning Outcomes Human Services. It provides support for infectious disease labora-
6. Compare and contrast the roles of the Centers for Disease Control tories in the United States and abroad and collects data on diseases
and Prevention, state public health departments, and the World that impact public health. Each week, the CDC publishes the
Health Organization. Morbidity and Mortality Weekly Report (MMWR), which summa-
7. Explain the value of maintaining a current list of notifiable diseases. rizes the status of a number of diseases. The MMWR is available
8. Describe the conditions that may allow eradication of a disease. online (http://www.cdc.gov/mmwr/), making it readily accessible
to anyone in the world.
Infectious disease surveillance, nationally and worldwide, is one The number of new cases of over 50 notifiable diseases is
of the most important aspects of disease prevention. It involves reported to the CDC by individual states (table 19.1). The list of

TABLE 19.1 Notifiable Infectious Diseases, 2011

Individual states and territories require physicians to report cases of these notifiable diseases. In turn, the number of cases is reported to the
CDC, where they are collated and published in the MMWR.
■ Anthrax ■ HIV infection ■ Shiga toxin–producing Escherichia coli (STEC)
■ Arboviral neuroinvasive and ■ Influenza-associated pediatric mortality ■ Shigellosis
non-neuroinvasive diseases ■ Legionellosis (Legionnaires’ disease) ■ Smallpox
■ Babesiosis ■ Listeriosis ■ Spotted fever rickettsiosis
■ Botulism ■ Lyme disease ■ Streptococcal toxic shock syndrome
■ Brucellosis ■ Malaria ■ Streptococcus pneumoniae, invasive disease
■ Chancroid ■ Measles ■ Syphilis
■ Chlamydia trachomatis, genital infections ■ Meningococcal disease ■ Tetanus
■ Cholera ■ Mumps ■ Toxic shock syndrome (other than
■ Coccidioidomycosis streptococcal)
■ Novel influenza A infections
■ Cryptosporidiosis ■ Trichinellosis (Trichinosis)
■ Pertussis
■ Cyclosporiasis ■ Tuberculosis
■ Plague
■ Dengue ■ Tularemia
■ Poliomyelitis, paralytic
■ Diphtheria ■ Typhoid fever
■ Poliovirus infection, nonparalytic
■ Ehrlichiosis/Anaplasmosis ■ Vancomycin-intermediate Staphylococcus
■ Psittacosis
■ Giardiasis aureus (VISA)
■ Q fever
■ Vancomycin-resistant Staphylococcus aureus
■ Gonorrhea ■ Rabies, animal and human (VRSA)
■ Haemophilus influenzae, invasive disease ■ Rubella ■ Varicella
■ Hansen’s disease (leprosy) ■ Rubella, congenital syndrome ■ Vibriosis
■ Hantavirus pulmonary syndrome ■ Salmonellosis ■ Viral hemorrhagic fevers
■ Hemolytic uremic syndrome, post-diarrheal ■ Severe acute respiratory syndrome– ■ Yellow fever
■ Hepatitis, viral, acute and chronic associated coronavirus (SARS-CoV) disease
 Part III Microorganisms and Humans 447

diseases considered notifiable is determined through collaborative 40 states have

efforts of the CDC and state health departments. Typically the health departments
1,000
diseases are of relatively high incidence or pose potential danger Influenza pandemic
to public health. The data collected by the CDC are published in
the MMWR along with historical numbers to reflect any trends. 800
Potentially significant case reports, such as the 1981 report of a Last human-to-human
cluster of opportunistic infections in young homosexual men that transmission of plague
600
heralded the AIDS epidemic, are also included in the MMWR. First use of penicillin

Rate
This publication is an invaluable aid to physicians, public health
agencies, teachers, students, and others concerned about infectious 400 Salk vaccine introduced
disease or public health. In fact, many of the epidemiological Passage of
charts and stories in this textbook are from the MMWR. First continuous Vaccination Assistance Act
200 municipal use
The CDC also conducts research relating to infectious dis- of chlorine in water
eases and can dispatch teams worldwide to assist with identifying in United States
and controlling epidemics. After all, an epidemic anywhere in the 0
world is a potential threat. In addition, the CDC provides refresher 1900 1920 1940 1960 1980 2000
courses for laboratory and infection control personnel. Year

MicroByte FIGURE 19.10 Crude Mortality Rate for Infectious Diseases,

Chlamydia, an STI, is the most commonly reported infectious disease United States, per 100,000 Population per Year
in the United States with over 1.2 million reported cases in 2009. ? What factors might cause an increase in the mortality rate over
the next two decades?

Public Health Departments

Each state has a public health laboratory responsible for infection The WHO disseminates information through a series of
surveillance and control as well as other health-related activities. periodicals and books. For example, the Weekly Epidemiological
Individual states have the authority to mandate which diseases must Record reports timely information about epidemics of public
be reported by physicians to the state laboratory. The prompt health importance, particularly those of global concern.
response of health authorities in Washington State that helped stop
an outbreak of Escherichia coli O157:H7 in 1993 caused by con- Reduction and Eradication of Disease
taminated hamburger patties was partly because Washington then Humans have been enormously successful in eliminating or reduc-
was one of the few states with surveillance and reporting measures ing the occurrence of certain diseases through efforts in improved
for the organism. The epidemic had actually started in other states sanitation, reservoir and vector control, vaccination, and antibiotic
but had gone unrecognized. Likewise, the fungus Cryptococcus treatment (figure 19.10). In the United States, many diseases that
gattii appeared in Washington State in 2007, causing lung infections were once common and claimed many lives are now relatively rare.
and meningitis in healthy persons, and is now closely watched by Successful vaccination programs have led to dramatic decreases
health authorities in Oregon and California. Cryptococcus gattii, p. 660 in the number of deaths caused by Haemophilus influenzae,
Corynebacterium diphtheriae, Clostridium tetani, and Bordetella
Other Components of the Public pertussis. Meanwhile, recognizing and controlling the source of
Health Network diseases such as malaria, plague, and cholera have been effective
The public health network also includes public schools, which in limiting their spread.
report absentee rates, and hospital laboratories, which report on One human disease—smallpox—
the isolation of pathogens with epidemiological significance. In has been globally eradicated, elimi-
conjunction with these local activities, the news media alert the nating the natural occurrence of
general public to the presence of infectious disease. a disease that had a 25% case-
fatality rate and disfigured many
who survived (figure  19.11).
Worldwide Disease Surveillance
The World Health Organization (WHO), an agency of
the United Nations with 193 member states, is devoted
to achieving the highest possible level of health around
the globe. It has four main functions: (1) provide world- FIGURE 19.11
Case of Smallpox, A
wide guidance in the field of health; (2) set global Disease That Has Now
standards for health; (3) cooperatively strengthen Been Eradicated
national health programs; and (4) develop and transfer
? What characteristics
appropriate health technology. To accomplish its goals, make a disease a
the WHO provides education and technical assistance to good candidate for
member countries. eradication?
448 Chapter 19 Epidemiology

Because humans were the only reservoir for the smallpox virus, pro- In the case of avian influenza, scientists are concerned that the
grams of extensive immunization along with isolation of cases were virus will evolve to spread from person to person. Resistance
able to eliminate the reservoir and therefore eradicate the disease. The to antimicrobial drugs is contributing to the reemergence of
WHO hopes to eradicate polio, measles, and dracunculiasis soon. many diseases, including malaria. avian flu, p. 509 malaria,
Political and social upheaval, complacency, and lack of financial sup- p. 686
port can result in a resurgence of diseases unless the pathogens are ■ Complacency and public health efforts. As an infectious
completely eliminated. polio, p. 656 measles, p. 537 disease is controlled and therefore of less concern, compla-
Scientific advances made over the past several decades led to cency can develop, paving the way for resurgence of the dis-
speculation that the war against infectious diseases, particularly ease. The early success of the plan to eliminate tuberculosis
those caused by bacteria, had been won. Microorganisms, how- in the United States by the year 2000 caused news reports,
ever, have occupied this planet far longer than humans, evolving education, and research money to be diverted to more com-
to reside in every habitat with the potential for life, including the mon diseases. Also, some social welfare programs were cur-
human body. It should be no surprise that new and previously tailed as the AIDS crisis demanded more funding. As a result,
unrecognized pathogens are emerging, and that some of those we poor health and living conditions put more people at risk of
had previously controlled are now making a comeback. developing active tuberculosis and the disease reemerged as
an increasing threat. Fortunately, new public health measures
MicroAssessment 19.3 have brought the disease back under control in the United
Across the United States, a network of agencies, including the Centers States (see figure 21.17). tuberculosis, p. 502
for Disease Control and Prevention (CDC) and state and local public ■ Changes in human society. Daycare centers, where diapered
health departments, monitors disease development. The World Health infants mingle, oblivious to sanitation and hygiene, are a rela-
Organization (WHO) is devoted to achieving the highest possible
tively new component of American society. For obvious reasons,
level of health for people around the globe. Humans have been
successful in reducing or eliminating certain diseases. these centers can be hotbeds of contagious diseases. This is par-
ticularly true for the intestinal pathogens Giardia and Shigella,
7. What is the MMWR?
which have a low infectious dose, because infants often explore
8. Explain why smallpox was successfully eradicated, but rabies through taste and touch and are thus likely to ingest fecal organ-
probably never will be.
isms (figure 19.12). Moreover, many young children have not
9. Explain why we have relatively accurate data on the number of yet acquired immunity to communicable diseases such as colds
cases of measles that occur in the United States but not on the
number of cases of the common cold. +
and diarrhea that are readily transmitted among this susceptible
population. Giardia, p. 602 Shigella, p. 588

■ Advances in technology. Technology can make life easier, but

can inadvertently create new habitats for microorganisms. The
19.4 ■ Emerging Infectious advent of contact lenses gave microorganisms the opportunity
Diseases to grow in a new location—the lenses and storage solutions of

Learning Outcomes
9. Explain two ways in which microbial evolution can lead to
emergence of disease.
10. Describe how human behavior can contribute to the emergence
and reemergence of disease.
11. Explain how climate can influence disease emergence.

Emerging diseases are those that are novel or have recently

increased in incidence. These include new or newly recognized
diseases such as severe acute respiratory syndrome (SARS),
mad cow disease, and avian flu, as well as familiar ones such as
malaria and tuberculosis that are reemerging after years of decline
(see figure  1.4). Microbes are adept at taking advantage of new
opportunities to thrive. Some of the factors that contribute to the
emergence and reemergence of diseases include:
■ Microbial evolution. The emergence of some diseases fol-
lows the natural evolution of microbes. For example, a new
serotype of Vibrio cholerae, designated O139, has gained the FIGURE 19.12 Children in a Daycare Center
ability to produce a protective capsule. Immunity against the ? Explain why Shigella and Giardia species are readily transmitted in
earlier strain does not guarantee immunity to the newer one. daycare centers.
 Part III Microorganisms and Humans 449

users who did not employ proper disinfection techniques. In 19.5 ■ Healthcare-Associated
turn, this resulted in new types of eye infections.
■ Population expansion. In a growing population humans
Infections
can expand into areas where they come into contact with Learning Outcomes
new reservoirs of disease (such as occurred with the Ebola
12. Describe four reservoirs of infectious agents in healthcare
virus), or have increased contact with established reservoirs. settings and three mechanisms by which the agents can be
As more homes are built along the borders of forests, for transferred to patients.
example, humans are exposed to more deer that carry ticks, 13. Describe the role of Infection Control Committees in preventing
the host for Borrelia burgdorferi, the cause of Lyme disease. nosocomial infections.
Lyme disease, p. 531

Healthcare-associated infections (HAIs), which are infec-

MicroByte
About 75% of emerging infectious diseases affecting humans are of
tions individuals acquire while receiving treatment in a health-
animal origin. care setting, are one of the top 10 causes of death in the United
States. Perhaps this should not be surprising—hospitals, in
■ Development. Dams, which provide important sources of particular, can be viewed as densely populated communities
power necessary for economic development, have inadvert- of unusually susceptible people where the most antimicrobial
ently extended the range of certain diseases. Transmission resistant and virulent pathogens can potentially circulate. In
of the disease schistosomiasis relies on the presence of an fact, hospital-acquired infections, or nosocomial infections,
aquatic snail that serves as a host for the Schistosoma parasite. have been a problem since hospitals began (nosocomial is
Construction of dams such as the Aswan Dam on the Nile derived from the Greek word for hospital). Modern medical
River has increased the habitat for the snail, thus extending practices, however, including the extensive use of antimicro-
the distribution of the disease. schistosomiasis, p. 350 bial drugs and invasive therapeutic procedures, have increased
the incidence of these infections. In the United States alone, an
■ Mass production, widespread distribution, and importation
estimated 5% to 10% of patients admitted to a hospital develop
of food. Foodborne illness has always existed, but the ease
a nosocomial infection, adding billions to the price of health-
with which we can now transport items worldwide from a
care. Many of these infections originate from the patient’s own
single production plant can create new problems. Widespread
normal microbiota, but approximately one-third are poten-
distribution of foods contaminated with pathogens can result
tially preventable. Figure 19.13 shows the relative frequency
in a similarly broad outbreak of disease. Contaminated
of different types of nosocomial infections.
hotdogs shipped from a single food manufacturing plant in
The severity of healthcare-associated infections can range
Canada in 2008 led to a country-wide epidemic of listeriosis
from very mild to fatal. Some infections arise during a hos-
that killed 21 people.
pital stay, but others are not discovered until after the patient
■ War and civil unrest. Wars and civil unrest can disrupt the has been discharged. Many factors influence the development
infrastructure on which disease prevention relies. Refugee of these infections such as the length of exposure, the man-
camps that crowd people into substandard living quarters ner in which a patient is exposed, the virulence and number
lacking toilet facilities and safe drinking water are hot- of organisms involved, and the immune state of the host.
beds of infectious diseases such as cholera and dysentery. Characteristics of some of the microorganisms and viruses
Unfortunately, war also disrupts disease eradication efforts. commonly implicated in healthcare-associated infections are
■ Climate changes. Warm temperatures favor the reproduction summarized in table 19.2.
and survival of some arthropods, which can serve as vectors
for diseases such as malaria and West Nile encephalitis. The
heavy rainfall and flooding caused by the effects of El Niño
may be related to surges of cholera cases in Africa.
Other Bacteremia
17% 14%
MicroAssessment 19.4
Microbes are adept at responding to change. Evolution of the
pathogen, changes in human behavior, or climate change can provide Surgical site Urinary tract
22% 32%
new opportunities for pathogens to thrive.
10. Why might a person immunized against cholera contract the Respiratory tract
disease anyway? 15%
11. How would you expect a trend toward warmer climates to affect
the spread of vector-borne disease?
FIGURE 19.13 Relative Frequency of Different Types of
12. What political and societal factors might lead to a decrease Nosocomial Infections
in childhood immunizations? +
? What is bacteremia?
450 Chapter 19 Epidemiology

TABLE 19.2 Common Causes of Healthcare-Associated Infections

Infectious Agent Comments

Acinetobacter baumannii This environmental bacterium can be found on skin of healthy people and is resistant to a number of
antimicrobial medications. It causes a variety of healthcare-associated infections including bloodstream and
surgical site infections and pneumonia.
Candida species These yeasts, part of the normal microbiota, are a common cause of healthcare-associated bloodstream
infections. Some are resistant to a number of antifungal medications. Candida albicans, p. 545

Clostridium difficile Toxin-producing strains of this bacterium can cause diarrhea and colitis in people taking antibiotics. Because the
bacterium produces endospores, which are not killed by disinfectants, thorough handwashing is an important
means of preventing transmission. Clostridium difficile-associated disease, p. 594

Enterococcus species These bacteria, part of the normal intestinal microbiota, are a common cause of nosocomial urinary tract
infections as well as wound and bloodstream infections. Some strains are resistant to all conventional
antimicrobial drugs. enterococci, p. 258

Escherichia coli and These members of the normal intestinal microbiota commonly cause healthcare-associated urinary tract and
other members of the other infections. Enterobacteriaceae, p. 265
Enterobacteriaceae
Norovirus This virus infects the gastrointestinal tract, causing vomiting and diarrhea. It has a very low infectious dose, so
scrupulous cleaning and handwashing are necessary to prevent its transmission. norovirus, p. 596

Pseudomonas species These bacteria grow readily in many moist, nutrient-poor environments such as the water in the humidifier of
a mechanical ventilator. Pseudomonas species are resistant to many disinfectants and antimicrobial drugs. They
are a common cause of healthcare-associated pneumonia, and infections of the urinary tract and burn wounds.
Pseudomonas, p. 554

Respiratory syncytial virus This virus is easily transmitted, and causes outbreaks of healthcare-associated lower respiratory tract infections,
(RSV) particularly at times when the virus is circulating in non-healthcare settings. respiratory syncytial virus, p. 511

Staphylococcus aureus Many people, including healthcare personnel, are carriers of this bacterium. Because it survives for prolonged
periods in the environment, it is readily transmissible on fomites. It is a common cause of healthcare-associated
pneumonia and surgical site infections. Hospital-acquired strains are often resistant to a variety of antimicrobial
drugs. Staphylococcus aureus, p. 524

Staphylococcus species These members of the normal skin microbiota can colonize the tips of intravenous catheters (small plastic tubes
other than S. aureus inserted into the veins). The resulting biofilms continuously seed organisms into the bloodstream and increase
the likelihood of a systemic infection. biofilm, p. 84

Reservoirs of Infectious Healthcare Environment

Agents in Healthcare Settings Some Gram-negative rods, particularly the common opportunis-
tic pathogen Pseudomonas aeruginosa, can thrive in healthcare
The organisms that cause healthcare-associated infections can
environments such as sinks, respirators, and toilets. Not only is
originate from a number of different sources, including other
P.  aeruginosa resistant to many disinfectants and antimicrobial
patients, the healthcare environment, healthcare workers, and
drugs, it requires few nutrients, enabling it to multiply in envi-
the patient’s own microbiota. Because of the widespread use of
ronments containing little other than water. Many nosocomial
antimicrobial drugs in hospitals, many organisms that cause noso-
infections have been traced to soaps, disinfectants, and other
comial infections such as methicillin-resistant Staphylococcus
aqueous solutions that were contaminated with the organism.
aureus (MRSA) are resistant to these medications.
Pseudomonas aeruginosa, p. 554

MicroByte
Healthcare Workers
In 2007, more people died in the United States from hospital-
acquired MRSA infections than from AIDS. Outbreaks of healthcare-associated infections are sometimes
traced to infected personnel. Clearly, those who report to work
with even a mild case of influenza can expose patients to an infec-
Other Patients tious agent that has serious or fatal consequences to those in poor
Because of the very nature of healthcare settings, infectious agents health. A more troublesome source of infection is a healthcare
are always present. In fact, patients are often hospitalized because worker who is a carrier of a pathogen such as Staphylococcus
they have a severe infectious disease. The pathogens that these aureus or Streptococcus pyogenes. These personnel may not
patients harbor can be discharged into the environment via skin recognize they pose a risk to patients until they are implicated in
cells, respiratory droplets, and other body secretions and excre- an outbreak. Carriers who are members of a surgical team pose a
tions. Scrupulous cleaning and the use of disinfectants minimize particular threat because inoculation of a pathogen directly into a
the spread of these pathogens. surgical site can result in a systemic infection.
 Part III Microorganisms and Humans 451

Patient Microbiota Fomite Transmission—Medical Devices

Many healthcare-associated infections originate from the patient’s Healthcare-associated infections most often result from medi-
own microbiota. Nearly any invasive procedure can transmit these cal devices that breach the first-line barriers of the normal host
organisms to otherwise sterile body sites. When intravenous flu- defense. Catheterization of the urinary tract can readily introduce
ids are administered, for example, Staphylococcus epidermidis, microorganisms into the normally sterile bladder. Because urine
a common member of the skin microbiota, can potentially gain is an excellent growth medium, the urinary tract often becomes
access to the bloodstream causing bacteremia. Although the infected. Urinary tract infections are the most common type of
immune system can usually eliminate these normally benign nosocomial infection (see figure 19.13).
organisms, the underlying illness of many hospitalized patients Just as urinary catheters can introduce bacteria into the blad-
compromises their immunity and they can develop a bloodstream der, intravenous (IV) catheters can introduce microorganisms into
infection. Patients who undergo intestinal surgery are prone to sur- the bloodstream. This can happen when normal skin microbiota
gical site infection by their normal bowel microbiota. Similarly, colonize the tip of an indwelling catheter or when environmental
patients who are on certain medications or have impaired cough organisms contaminate IV fluids or the lines that deliver them.
reflexes can inadvertently inhale their normal oral microbiota, Even normally benign skin microbiota can cause life-threatening
resulting in healthcare-associated pneumonia. bacteremia, p. 384 bacteremia if they gain access to the bloodstream.
Severely immunocompromised patients, such as people Mechanical respirators that assist a patient’s breathing by
who have undergone cancer chemotherapy or are on immuno- pumping air directly into the trachea can potentially deliver micro-
suppressive drugs, are prone to activation of latent infections their organisms. For example, if a nutritionally versatile organism such
immune system could previously control. For example, latent as a Pseudomonas species gains access to water droplets in the
infections of Toxoplasma gondii, a protozoan parasite commonly machine, it can multiply and be pumped into the lungs.
acquired during childhood, can become activated and cause a life- Inadequately sterilized instruments that are used in invasive
threatening disease. latent infection, p. 322 procedures such as surgery or biopsy can also transmit infectious
agents. Endoscopes and other heat-sensitive instruments are often
Transmission of Infectious treated with chemical sterilants to render them microbe-free.
Agents in Healthcare Settings Improper use of these chemicals, however, can result in the sur-
vival of some organisms. sterilants, p. 116
Diagnostic and therapeutic procedures can potentially transmit
infectious agents to patients. This is particularly true in intensive Direct Transmission—Healthcare Personnel
care units (ICUs), where patients generally have indwelling cath-
Healthcare personnel must be extremely vigilant to avoid trans-
eters used to deliver intravenous fluids or monitor the patient’s
mitting infectious disease agents, particularly from patient to
condition (figure 19.14).
patient. What Ignaz Semmelweis found to be true in the 1800s is
equally true today—handwashing between contact with individual
patients helps prevent the spread of disease (see A Glimpse of
History). Unfortunately, this relatively simple procedure is too
often overlooked.
Healthcare personnel should routinely wash or disinfect their
hands after touching one patient before going to the next. They
should perform a more thorough hand scrubbing lasting 10 min-
utes with a strong disinfectant before participating in a surgical
operation, or when working in a nursery, or an intensive care or
isolation unit. Healthcare workers are instructed to wear gloves
when they have contact with blood, mucous membranes, broken
skin, or body fluids.

Airborne Transmission
Most hospitals are designed to minimize the airborne spread
of microorganisms. Airflow to operating rooms is usually
regulated so that it is supplied under slight pressure, thereby
preventing contaminated air in the corridors from flowing into
the room. Floors are washed with a damp mop or floor washer
rather than swept in order to avoid resuspending microbes into
the air. To exclude airborne microorganisms and viruses from
rooms in which extremely susceptible patients reside, such as
those who have recently undergone a bone marrow transplant,
high-efficiency particulate air (HEPA) filters are used. These
FIGURE 19.14 Patient in an Intensive Care Unit filter out most airborne particles, including microorganisms.
? To what sources of nosocomial infections is this patient exposed? HEPA filters, p. 115
452 Chapter 19 Epidemiology

PERSPECTIVE 19.1
Standard Precautions—Protecting Patients and Healthcare Personnel
One of the biggest challenges for a hospital has guidelines were updated and expanded in mouth/nose when sneezing/coughing;
always been to prevent spread of disease within 2007 to include recommendations that can be use tissues and dispose in a no-touch
that confined setting. Patients with infectious applied to all healthcare facilities. The advi- receptacle; observe hand hygiene
diseases were once in separate hospitals; those sory committee has also been renamed, sub- after soiling of hands with respiratory
with similar diseases were sometimes housed stituting the word “healthcare” for “hospital,” secretions; wear surgical mask if tolerated
in clusters on the same floor. In 1910, a cubicle to reflect the expanded scope of the infection or maintain spatial separation of more
system of isolation was introduced in which control recommendations. than 3 feet if possible.
patients were placed in multiple-bed wards. The current guidelines have two tiers of ■ Patient placement. Prioritize for single-
Aseptic nursing procedures were aimed at pre- isolation procedures: The fundamental mea- patient room if patient is at increased risk
venting the transmission of infectious agents to sures are the Standard Precautions, designed of transmission, is likely to contaminate
other patients and personnel. These scrupulous for the care of all patients. The Transmission- the environment, does not maintain
measures were so successful that general hospi- Based Precautions are supplementary mea- appropriate hygiene, or is at increased
tals were able to accommodate infectious dis- sures to be used in addition to the Standard risk of acquiring infection or developing
ease patients, ultimately resulting in the closure Precautions if a patient is, or might be, infected adverse outcome following infection.
of many infectious disease hospitals beginning with a highly transmissible or epidemiologi- ■ Patient-care equipment and
in the 1950s. To help general hospitals with cally important pathogen. These are separated instruments/devices. If the equipment is
isolation procedures, in 1970 the CDC began into three sets—Airborne Precautions, Droplet soiled, handle in a manner that prevents
publishing a manual that recommended a cat- Precautions, and Contact Precautions—that are transfer of microorganisms to others and
egory system of seven isolation procedures for used singly or in combination as appropriate. to the environment; wear gloves if visibly
patients based on their diagnosis. The Standard Precautions can be sum- contaminated; perform hand hygiene.
In the early 1980s, healthcare workers marized as:
■ Care of the environment. Develop
began to acquire hepatitis B and, later, HIV
■ Hand hygiene. During the delivery of procedures for routine care, cleaning, and
from contact with the blood or other body
healthcare, avoid unnecessary touching disinfection of environmental surfaces,
fluids of patients. Existing guidelines were
of surfaces in close proximity to the especially frequently touched surfaces in
primarily aimed at preventing patient-to-
patient. When hands are visibly dirty or patient-care areas.
patient transmission of disease, rather than
patient-to-personnel. In response, the CDC contaminated, wash them with soap and ■ Textiles and laundry. Handle in
recommended an additional set of guide- water; if non-antimicrobial soap is used, a manner that prevents transfer of
lines, the Universal (Blood and Body Fluid) decontaminate hands with an alcohol- microorganisms to others and to the
Precautions, to be followed when working based hand rub. If hands are not visibly environment.
with any patient, regardless of the diagno- soiled, decontaminate hands before direct ■ Safe injection practices. Use aseptic
sis. These defined the situations in which contact with patients; after contact with technique to avoid contamination of
gloves, gowns, masks, and eye protection were blood, body fluids, secretions, excretions, sterile injection equipment. Specific
required to prevent contact with blood. Many contaminated items; immediately after precautions describe how medications and
hospitals then broadened this concept, requir- removing gloves; and between patient IV solutions are stored and administered.
ing the use of gloves to isolate all moist and contacts. If contact with spores is likely to
■ Infection control practices for special
potentially infectious body substances. This have occurred, wash with soap and water.
lumbar puncture procedures. Wear a
approach, Body Substance Isolation, made ■ Personal protective equipment. Gloves surgical mask when placing a catheter or
the traditional diagnosis-dependent isolation are worn when touching blood, body fluids, injection material into the spinal canal or
procedures largely unnecessary. secretions, excretions, and contaminated subdural space.
The advent of Universal Precautions and items; and for touching mucous membranes
■ Worker safety. Adhere to federal and
Body Substance Isolation, in addition to the and non-intact skin. A gown is worn during
state requirements for protection of
previous diagnosis-dependent isolation pro- procedures and patient-care activities
healthcare personnel from exposure to
cedures, resulted in such a mix of recom- when contact of clothing/exposed skin
bloodborne pathogens.
mendations that it generated a great deal of with blood/body fluids, secretions, and
confusion. No existing, single set of guidelines excretions is anticipated. Mask, goggles, From Siegel, J.D., Rhinehart, E., Jackson,
was sufficient, and it was not clear when each or a face shield is worn during procedures M., Chiarello, L., and the Healthcare Infection
should be used. In response, the CDC and the and patient-care activities likely to generate Control Practices Advisory Committee,
Hospital Infection Control Practices Advisory splashes or sprays of blood, body fluids, or 2007 Guidelines for Isolation Precautions:
Committee (HICPAC) established a new set secretions. Preventing Transmission of Infectious Agents
of guidelines in 1996 that incorporated the ■ Respiratory hygiene/cough etiquette. in Healthcare Settings, June 2007. http://www
strengths of each of the alternatives. The Instruct symptomatic persons to cover .cdc.gov/ncidod/dhqp/pdf/isolation2007.pdf

Preventing Healthcare-Associated policies to prevent their development. To accomplish this, nearly

Infections every hospital has an Infection Control Committee, composed
The most important steps in preventing healthcare-associated of representatives of the various professionals in the hospital,
infections are to first detect their occurrence and then establish including nurses, physicians, dietitians, housekeeping staff, and
 Part III Microorganisms and Humans 453

microbiology laboratory personnel. On this committee, and some- MicroAssessment 19.5

times chairing it, is often a hospital epidemiologist, a professional
Healthcare-associated infections may originate from other
specially trained in hospital infection control. Hospitals may
patients, the healthcare environment, healthcare workers, or the
employ an infection control practitioner (ICP), whose role is to patient’s own normal microbiota. Diagnostic and therapeutic
perform active surveillance of the types and numbers of infections procedures can potentially transmit infectious agents. The most
that arise in the hospital. The Infection Control Committee, in con- important steps in preventing healthcare-associated infections
junction with the ICP, drafts and implements preventive policies are to first detect their occurrence and then establish policies to
following the guidelines suggested by the Standard Precautions prevent their development.
and the Transmission-Based Precautions (see Perspective 19.1). 13. Explain why an IV catheter poses a risk to a patient.
The CDC also takes an active role in preventing healthcare- 14. Describe two ways in which infectious agents can be transmitted
associated infections, and has established the Healthcare Infection to a patient.
Control Practices Advisory Committee (HICPAC). The role of 15. The rate of nosocomial infections is often relatively high in
this national committee is to provide advice to hospitals and rec- emergency room settings. Explain why this might be so. +
ommend guidelines for surveillance, prevention, and control of
healthcare-associated infections.

FUTURE CHALLENGES 19.1

Maintaining Vigilance Against Bioterrorism
Today, an unfortunate challenge in epide- infection. Gastrointestinal anthrax results by viruses, effective drug therapy is not
miology is to maintain vigilance against from consuming contaminated food, available. Special isolation precautions
bioterrorism—the deliberate release of infec- leading to vomiting of blood and severe must be used for smallpox patients
tious agents or their toxins as a means to cause diarrhea; it is not common but has a high because the virus can be acquired through
harm. Even as we work to control diseases, case-fatality rate. Anthrax can be droplet, airborne, or contact transmission.
we must be aware that microbes pose a threat prevented by vaccination, but that option ■ Francisella tularensis. This bacterium,
as agents of bioterrorism. Hopefully, future is not widely available. Prophylaxis with naturally found in animals such as
attacks will never occur, but it is crucial antimicrobial medications is possible rodents and rabbits, causes the disease
to be prepared for the possibility. Prompt for those who might have been exposed, tularemia. Inhalation of the bacterium
recognition of such an event, followed by but this requires prompt recognition results in severe pneumonia, which is
rapid and appropriate isolation and treatment of exposure. Fortunately, person-to- incapacitating but would probably have a
procedures, can help to minimize the con- person transmission of the agent is lower case-fatality rate than inhalational
sequences. The CDC, in cooperation with not likely. anthrax or plague. A vaccine is not
the Association for Professionals in Infection ■ Botulism. Botulism is caused naturally available, but post-exposure prophylaxis
Control and Epidemiology (APIC), has pre- by the ingestion of botulinum toxin, with antimicrobial medications is
pared a bioterrorism readiness plan to be used produced by Clostridium botulinum. Any possible. Fortunately, person-to-person
as a template by healthcare facilities. Many mucous membrane can absorb the toxin, transmission of the agent is not likely.
of the recommendations are based on the so aerosolized toxin could be used as a ■ Viruses that cause hemorrhagic fevers.
Standard Precautions already used by hospitals weapon. Botulism can be prevented by These include various viruses such as
to prevent the spread of infectious agents (see vaccination, but that option is not widely Ebola and Marburg. Symptoms vary
Perspective 19.1). available. An antitoxin is also available depending on the virus, but severe cases
The CDC separates bioterrorism agents in limited supplies. Botulism is not show signs of bleeding from many sites.
into three categories based on the ease of contagious. There are no vaccines against these
spread and severity of disease. Category A ■ Yersinia pestis. Pneumonic plague, viruses, and generally no treatment.
agents pose the highest risk because they are caused by inhalation of Yersinia pestis, Some, but not all, of these viruses can
easily spread or transmitted from person to is the most likely form of plague to result be transmitted from person to person,
person and result in high mortality. from a biological weapon. Although so patient isolation in these cases is
These agents include: no effective vaccine is available, post- important.
■ Bacillus anthracis. Endospores of this exposure prophylaxis with antimicrobial
bacterium were used in the bioterrorism medications is possible. Special isolation Category B agents pose moderate risk because
events of 2001. The most severe outcome, precautions must be used for patients they are relatively easy to spread and cause mod-
inhalational anthrax, results when an who have pneumonic plague because the erate morbidity. These agents include organisms
individual breathes in the airborne spores. disease is easily transmitted by respiratory that cause food- and waterborne illness, various
It can lead to a rapidly fatal systemic droplets. biological toxins, Brucella species, Burkholderia
illness. Cutaneous anthrax, which occurs ■ Smallpox. Although a vaccine is mallei and pseudomallei, Coxiella burnetii, and
when the organism enters the skin, available to prevent infection with this Chlamydophila (Chlamydia) psittaci. Category
manifests as a blister that develops into a virus, routine immunization was stopped C agents are emerging pathogens that could be
skin ulcer with a black center. Although over 30 years ago because the natural engineered for easy dissemination. These include
this usually heals without treatment, it disease has been eradicated. As is the Nipah virus, which was first recognized in 1999,
can also progress to a fatal bloodstream case with nearly all infections caused and hantavirus, first recognized in 1993.
454 Chapter 19 Epidemiology

Summary
19.1 ■ Principles of Epidemiology from a propagated epidemic (figure 19.8). Some epidemics are seasonal
Epidemiologists study the frequency and distribution of disease in order (figure 19.9).
to identify its cause, source, and route of transmission.
Analytical Studies
Rate of Disease in a Population Analytical studies try to determine which risk factors are actually
Epidemiologists focus on the rate of disease. Diseases that are con- relevant to disease development. A retrospective study compares the
stantly present in a population are endemic; an unusually large number activities of cases with controls to determine the cause of the epidemic.
of cases in a population constitutes an epidemic (figure 19.1). A prospective study compares cohort groups, to determine if the iden-
tified risk factors predict a tendency to develop disease.
Reservoirs of Infection (figure 19.2)
Preventing susceptible people from coming in contact with a reservoir Experimental Studies
of infection can prevent infectious disease. People who have asymp- Experimental studies are generally used to evaluate the effectiveness
tomatic infections or are colonized with a pathogen are carriers of the of a treatment or intervention in preventing disease.
infectious agent. Zoonoses such as plague and rabies can be transmitted
to humans but exist primarily in other animals. Pathogens with environ- 19.3 ■ Infectious Disease Surveillance
mental reservoirs are probably impossible to eliminate.
National Disease Surveillance Network
Portals of Exit and Portals of Entry The Centers for Disease Control and Prevention (CDC) collects data
Pathogens may be shed in feces, in respiratory droplets, on skin cells, in on diseases of public health importance and summarizes their status in
genital secretions, and in urine. Many organisms can cause disease if they the Morbidity and Mortality Weekly Report (MMWR); other activities of
enter one body site, but are harmless if they enter another (figure 19.3). the CDC include research, assistance in controlling epidemics, and sup-
port for infectious disease laboratories. State public health departments
Disease Transmission (figure 19.4)
are involved in infection surveillance and control (table 19.1).
Handwashing is a key control measure in preventing diseases that are
spread through direct or indirect contact, as well as those that spread via Worldwide Disease Surveillance
contaminated food. Direct contact occurs when one person physically The World Health Organization (WHO) is devoted to achieving the
touches another. Indirect contact involves transfer of pathogens via highest possible level of health for people around the globe.
fomites. Droplet transmission of respiratory pathogens is considered
Reduction and Eradication of Disease (figure 19.10)
contact because of the close proximity involved. Foodborne pathogens
can originate from the animal reservoir or from contamination during Smallpox has been eradicated (figure  19.11). The WHO hopes to soon
food preparation. Waterborne pathogens often originate from sewage eliminate polio, measles, and dracunculiasis.
contamination. Airborne transmission of pathogens is the most dif-
ficult to control (figure  19.5). A mechanical vector carries a microbe 19.4 ■ Emerging Infectious Diseases
on its body from one place to another. Pathogens can multiply to high Emerging diseases are new or newly recognized, or are reemerging
numbers in a biological vector (figure 19.6b). Prevention of vector-borne after years of decline. Factors that contribute to the emergence and
disease relies on vector control. reemergence of diseases include microbial evolution, the breakdown
of public health infrastructure, changes in human behavior, advances in
Pathogen Factors That Influence the Epidemiology technology, population expansion, economic development, mass distri-
of Disease bution and importation of food, war, and climate changes (figure 19.12).
Pathogens with a variety of virulence factors are more likely to cause
severe disease. The probability of infection and disease is generally 19.5 ■ Healthcare-Associated Infections
lower if an individual is exposed to small numbers of pathogens. Healthcare-associated infections (HAIs) are acquired by individuals
Diseases with a long incubation period can spread before the first cases in a healthcare setting. Nosocomial infections are acquired in a hospital
appear. (figure 19.13).

Host Factors That Influence the Epidemiology of Disease Reservoirs of Infectious Agents in Healthcare Settings
A disease is unlikely to spread very widely in a population showing The organisms that cause healthcare-associated infections may originate
herd immunity in which most people are immune to the disease agent. from other patients, the healthcare environment, healthcare workers, or
Malnutrition, overcrowding, and fatigue increase the susceptibility of the patient’s own microbiota.
people to infectious diseases. The very young and the elderly are gener-
ally more susceptible to infectious agents (figure 19.7). Natural immunity Transmission of Infectious Agents in Healthcare Settings
can vary with genetic background, but it is difficult to determine the Healthcare-associated infections can result from medical devices that
relative importance of genetic, cultural, and environmental factors. breach the first-line barriers of the normal host defense (figure  19.14).
Healthcare personnel should routinely wash or disinfect their hands
19.2 ■ Epidemiological Studies after touching one patient before going to the next.
Descriptive Studies Preventing Healthcare-Associated Infections
Descriptive studies attempt to identify potential risk factors that cor- The most important steps in preventing healthcare-associated infections
relate with the development of disease. Determining the time that are to first recognize their occurrence and then establish policies to
the illness occurred helps distinguish a common-source epidemic prevent both their development and spread.
 Part III Microorganisms and Humans 455

Review Questions
Short Answer
1. Describe the impact on a society of high incidence and high 5. Which of the following statements is false?
prevalence of an endemic debilitating disease. a) A disease with a long incubation period might spread
2. What is the epidemiological significance of people who have extensively before an epidemic is recognized.
asymptomatic infections? b) A person exposed to a low dose of a pathogen might not
develop disease.
3. Explain why zoonotic diseases are often severe in humans.
c) The young and the aged are more likely to develop certain
4. List the main portals of exit from the human body.
diseases.
5. Name the most important control measure for preventing person- d) Malnourished populations are more likely to develop
to-person transmission of a disease. certain diseases.
6. Describe the factors within a population that may make it more e) Herd immunity occurs when a population does not engage
susceptible to infectious disease. in a given behavior, such as eating raw fish, that would
7. Draw a representative graph (time versus number of people ill) otherwise increase their risk of disease.
depicting both a propagated and a common-source epidemic. 6. The purpose of an analytical study is to
8. Describe the differences between a retrospective (case-control) a) identify the person, place, and time of an outbreak.
study and a prospective (cohort) study. b) identify risk factors that result in high frequencies of
9. What information is available in the Weekly Epidemiological disease.
Record? c) assess the effectiveness of preventive measures.
10. Explain how smallpox was eradicated. d) determine the effectiveness of a placebo.
11. Describe the factors that contribute to the emergence or re- e) None of the above
emergence of disease. 7. Which of the following causes of emerging diseases is thought
12. What are the main reservoirs of nosocomial infections? to be a new pathogen?
a) Giardia b) Vibrio cholerae O139
Multiple Choice
c) Mycobacterium tuberculosis d) Shigella dysenteriae
1. Which of the following is an example of a fomite? e) Schistosoma
a) Table b) Flea c) Staphylococcus aureus carrier 8. All of the following are thought to contribute to the emergence
d) Water e) Air of disease except
2. Which of the following would be the easiest to eradicate? a) advances in technology.
a) A pathogen that is common in wild animals but sometimes b) breakdown of public health infrastructure.
infects humans c) construction of dams.
b) A disease that occurs exclusively in humans, always d) mass distribution and importation of food.
resulting in obvious symptoms e) widespread vaccination programs.
c) A mild disease of humans that often results in no obvious 9. Which of the following common causes of healthcare-associated
symptoms infections is an environmental organism that grows readily in
d) A pathogen found in marine sediments nutrient-poor solutions?
e) A pathogen that readily infects both wild animals and a) Enterococcus b) Escherichia coli
humans
c) Pseudomonas aeruginosa d) Staphylococcus aureus
3. Which of the following methods of disease transmission is the
10. What is the most common type of nosocomial infection?
most difficult to control?
a) Bloodstream infection b) Gastrointestinal infection
a) Airborne b) Foodborne c) Waterborne
c) Pneumonia d) Surgical wound infection
d) Vector-borne e) Direct person-to-person
e) Urinary tract infection
4. Which of the following statements is false?
a) A botulism epidemic that results from improperly canned Applications
green beans is an example of a common-source outbreak. 1. A news station reported about a potentially fatal epidemic disease
b) Droplet nuclei fall quickly to the ground. occurring in a small Laotian village. An epidemiologist from the
c) Congenital syphilis is an example of a disease acquired CDC was interviewed to discuss the disease and was very dis-
through vertical transmission. tressed that it was not being contained. Why did the epidemiologist
d) Plague is endemic in the prairie dog population in parts of feel the disease was a concern for people in North America?
the United States. 2. An international team was gathered to discuss how funding should
e) The first case in an outbreak is called the index case. be spent to eliminate human infectious disease. There is only enough
456 Chapter 19 Epidemiology

funding to eliminate one disease. How would the scientists go about 2. A student disagreed with the presentation of the examples in fig-
choosing the next disease to be eliminated from the planet? ure 19.8. She claimed that the number of cases from a common-
source outbreak could remain high over a much longer period of
Critical Thinking + time in some cases and not decrease to zero. Is the student’s claim
1. Yersinia pestis and hantavirus are both found in wild rodents in reasonable? Why or why not?
the southwestern United States. What is the risk of trying to stop a
hantavirus epidemic by destroying rodents in that region?
20 Antimicrobial Medications
KEY TERMS
Acquired Resistance Resistance
that develops through mutation or
acquisition of new genes.
Antibiotic A compound naturally
Broad-Spectrum Antimicrobial
An antimicrobial drug that is
effective against a wide range of
microorganisms, often including
both Gram-positive and Gram-
produced by molds or bacteria that
negative bacteria.
inhibits the growth of or kills other
microorganisms. Chemotherapeutic Agent A
chemical used to treat disease.
Antimicrobial Drug A chemical
that inhibits the growth of or Intrinsic (Innate) Resistance
kills microorganisms; the term Resistance due to inherent
encompasses antibiotics and characteristics of the organism.
chemically synthesized drugs. Narrow-Spectrum Antimicrobial
Antiviral Drug A drug that An antimicrobial drug that is
interferes with the infection cycle effective against a limited range of
of a virus. microorganisms.
Bactericidal Kills bacteria. R Plasmid A plasmid that
encodes resistance to one or more
Bacteriostatic Inhibits the growth
antimicrobial drugs.
of bacteria.

Antibiotic susceptibility testing.

A Glimpse of History effective in treating the disease in laboratory animals. Although the drug
itself was potentially lethal for patients, it did cure infections previously
Paul Ehrlich (1854–1915), a German physician and bacteriologist, was
considered hopeless. The drug was given the name Salvarsan, a term
born into a wealthy family. He was the only son after many daughters,
derived from the words salvation and arsenic. Ehrlich’s discovery proved
so family and servants indulged his interests, even though his large col-
that some chemicals could indeed selectively kill microbes.
lection of frogs and snakes occasionally entered the laundry room. As
an adult, he was rarely without a good cigar and habitually scribbled
notes on his shirt cuffs. After receiving a degree in medicine, he became

T
intrigued with the way various types of body cells differ in their ability hink back to the last time you were prescribed an anti-
to take up dyes and other substances. When he observed that certain dyes microbial medication. Could you have recovered from the
stain bacterial cells but not animal cells, indicating that the two cell types infection without the drug? The prognosis for people with
are somehow fundamentally different, it occurred to him that it might be common diseases such as bacterial pneumonia and severe staphy-
possible to find a chemical that selectively harms bacteria without affect- lococcal infections was grim before the discovery and widespread
ing human cells. availability of penicillin in the 1940s. Physicians were able to
Ehrlich began searching for a “magic bullet,” a term he used to identify the cause of the disease, but the only treatment option
describe a drug that would kill a microbial pathogen without harming was usually bedrest. Today, however, antimicrobials are rou-
the human host. He began by looking for a chemical that would cure
tinely prescribed, and this simple cure is often taken for granted.
the sexually transmitted disease syphilis, which is caused by the spiro-
Unfortunately, the misuse of these life-saving medications,
chete Treponema pallidum. Much of the mental illness during this time
resulted from tertiary syphilis, a late stage of the disease. Ehrlich knew coupled with the amazing ability of microorganisms to adapt, has
that an arsenic compound had shown some success in treating a proto- led to an increase in the number of drug-resistant organisms. Some
zoan disease of animals, and so he and his colleagues began synthesizing people even speculate that we are in danger of seeing an end to
hundreds of different arsenic compounds in search for a cure for syphilis. the era of antimicrobial medications. In response, scientists are
In 1910, the 606th compound tested, arsphenamine, proved to be highly scrambling to develop new drugs.

457
458 Chapter 20 Antimicrobial Medications

20.1 ■ History and Development World War II spurred cooperation of British and American scien-
tists to determine the chemical structure of penicillin and to
of Antimicrobial Drugs develop the means for its large-scale production. Several different
penicillins were found in the Penicillium cultures, and were desig-
Learning Outcomes nated alphabetically. Penicillin G (or benzyl penicillin) was found
1. Describe the discovery of antimicrobial drugs and antibiotics. to be the most suitable for treating infections. This was the first
2. Explain how new generations of antimicrobial drugs are of what we now call antibiotics—antimicrobial drugs naturally
developed. produced by microorganisms.
Soon after the discovery of penicillin, Selman Waksman iso-
To appreciate the unique antibiotic era in which we now live, it lated a bacterium from soil, Streptomyces griseus, that produced
is important to understand the history and development of these an antibiotic he called streptomycin. The realization that bacteria
life-saving remedies. as well as molds could produce antibiotics prompted researchers
to begin screening hundreds of thousands of different microbial
Discovery of Antimicrobial Drugs strains for antibiotic production. Even today, pharmaceutical com-
The development of Salvarsan by Paul Ehrlich was the first panies examine soil samples from around the world for microbes
documented example of a chemical used successfully as an that produce novel antibiotics.
antimicrobial medication (see A Glimpse of History). The next
breakthrough came almost 25 years later, when the German chem- Development of New
ist Gerhard Domagk discovered that a red dye called Prontosil Generations of Drugs
could be used to treat streptococcal infections in animals. In the 1960s, scientists discovered they could alter the chemical
Surprisingly, Prontosil had no effect on streptococci growing in structure of certain antimicrobial drugs, giving them new properties.
test tubes. It was later discovered that enzymes in the blood of the For example, penicillin G, which is active mainly against Gram-
animal split the Prontosil molecule, producing a smaller molecule positive bacteria, can be altered to produce ampicillin, a drug that
called sulfanilamide; this breakdown product acted against the kills a variety of Gram-negative species as well. Other changes to
infecting streptococci. Thus, the discovery of sulfanilamide, the penicillin created methicillin, which is less susceptible to enzymes
first sulfa drug, was based on luck as well as scientific effort. If used by some bacteria to destroy penicillin. Today a variety of
Prontosil had been screened only against bacteria in test tubes and penicillin-like medications exist, making up what is referred to as
not given to infected animals, its effectiveness might never have the family of penicillins (figure 20.1). Other unrelated antimicro-
been discovered. bial drugs have also been altered to give them new characteristics.
Salvarsan and Prontosil are chemotherapeutic agents, mean-
ing chemicals used to treat disease. Because they are used to treat MicroAssessment 20.1
microbial infections, they can also be called antimicrobial drugs
or, more simply, antimicrobials. Antimicrobials are chemotherapeutic agents that are effective in
treating microbial infections. Antibiotics are antimicrobial chemicals
naturally produced by particular microorganisms.
Discovery of Antibiotics 1. How is the microbe that makes penicillin different from the one
In 1928, Alexander Fleming, a British scientist, was working with that makes streptomycin?
cultures of Staphylococcus aureus when he noticed that colonies 2. Define and contrast the terms chemotherapeutic agent,
growing near a contaminating mold looked as if they were dissolv- antimicrobial drug, and antibiotic.
ing. Recognizing that the mold might be secreting a substance that 3. How might Streptomyces griseus cells protect themselves from
killed bacteria, he proceeded to study it more carefully. He identi- the effects of streptomycin? +
fied the mold as a species of Penicillium and found it was indeed
producing a bacteria-killing substance; he called this penicillin.
Even though Fleming was unable to purify penicillin, he showed 20.2 ■ Features of
that it was remarkably effective in killing many different bacterial Antimicrobial Drugs
species and did not cause adverse effects when injected into rab-
bits and mice. Fleming recognized the potential medical signifi- Learning Outcome
cance of his discovery, but became discouraged with his inability 3. Describe selective toxicity; antimicrobial action; spectrum of
to purify the compound and eventually abandoned his study of it. activity; tissue distribution/metabolism/excretion; effects of
About 10 years after Fleming’s discovery of penicillin, two combinations; adverse effects; and resistance to antimicrobials.
other scientists in Britain, Ernst Chain and Howard Florey, suc-
cessfully purified the compound. In 1941, the drug was tested Most modern antibiotics come from microorganisms that nor-
for the first time on a police officer with a life-threatening mally reside in the soil, including species of Streptomyces and
Staphylococcus aureus infection. The patient improved so dra- Bacillus (bacteria), and Penicillium and Cephalosporium (fungi).
matically that within 24 hours his illness seemed under control. To commercially produce an antibiotic, a carefully selected strain
Unfortunately, the supply of purified penicillin ran out, and the of the appropriate species is inoculated into a broth medium and
man eventually died of the infection. Later, with greater supplies incubated in a huge vat. As soon as the maximum antibiotic con-
of the drug, two deathly ill patients were successfully cured. centration is reached, the drug is extracted from the medium and
 Part III Microorganisms and Humans 459

1980 Temocillin this by interfering with essential structures or biochemical proc-

esses that are common in microbes but not human cells.
Although the ideal antimicrobial drug is non-toxic to humans,
Mezlocillin most can be harmful at high concentrations. In other words, selec-
tive toxicity is a relative term. The toxicity of a given drug is
Piperacillin
expressed as the therapeutic index, which is the lowest dose
Apalcillin Talampicillin toxic to the patient divided by the dose typically used for
Carfecillin
1975 Azlocillin therapy. Antimicrobials that have a high therapeutic index
Pivmecillinam are less toxic, often because the drug acts against a vital
Bacampicillin biochemical process of microorganisms that does not
exist in human cells. For example, penicillin G, which
Carindacillin interferes with bacterial cell wall synthesis, has a very
Sulbenicillin Epicillin high therapeutic index.
Mecillinam
Cyclacillin When an antimicrobial that has a low therapeutic
1970 Ticarcillin
Amoxicillin index is used, the concentration in the patient’s blood
Flucloxacillin
must be carefully monitored to make sure it does not
Azidocillin Pivampicillin
reach a toxic level. Drugs too toxic for systemic use
Carbenicillin can sometimes be used for topical applications, such as
first-aid antibiotic skin ointments.
1965

Dicloxacillin Antimicrobial Action

Cloxacillin Some antimicrobial drugs usually kill microbes, whereas
Ampicillin others only inhibit their growth. Both actions are medically
Oxacillin
Propicillin Nafcillin
important.
1960 Bacteriostatic drugs inhibit bacterial growth. A patient
Phenethicillin
Methicillin taking a bacteriostatic drug must rely on his or her body’s
6-APA defense systems to kill or eliminate the pathogen after its growth
1955 has been stopped. Sulfa drugs, for example, are frequently pre-
scribed for urinary tract infections. They prevent bacteria in the
Penicillin V bladder from growing, so that urination can more effectively
1950
eliminate them.
Penicillin G Bactericidal drugs kill bacteria. These are particularly
useful when the host defenses cannot be relied on to eliminate
Penicillium chrysogenum pathogens. Bactericidal drugs are sometimes only inhibitory,
FIGURE 20.1 Family Tree of Penicillins All of the derivatives depending on the drug concentrations and the stage of bacterial
contain 6-aminopenicillanic acid (6-APA), the core portion of growth. Surprisingly, the lethal effects of different bactericidal
penicillin G. drugs seem to involve the same mechanism, regardless of the
? Why is it necessary to develop new generations of antimicrobial drug’s target. It appears that the antimicrobial-induced cell
medications? damage overwhelms the bacterium’s ability to detoxify reac-
tive oxygen species, leading to extensive oxidative damage.
reactive oxygen species, p.  90
extensively purified. In many cases, the antibiotic is chemically
altered after purification to give it new characteristics such as
increased stability. These chemically modified compounds are Spectrum of Activity
called semisynthetic. In some cases, the entire drug can be syn- Antimicrobial drugs vary with respect to the range of microorgan-
thesized in the laboratory. By convention, these partially or totally isms they kill or inhibit. Some kill or inhibit a narrow range of
synthetic chemicals are still called antibiotics because micro- microorganisms, such as only Gram-positive bacteria, whereas
organisms can produce the core structure naturally. Numerous others affect a wide range, generally including both Gram-positive
different antimicrobial drugs are now available, each with char- and Gram-negative organisms.
acteristics that make it more or less suitable for a given clinical Broad-spectrum antimicrobials affect a wide range of bac-
situation. Hundreds of tons and many millions of dollars’ worth of teria. These drugs are important for treating acute life-threatening
antibiotics are now produced each year. diseases when immediate antimicrobial therapy is essential and
there is no time to culture and identify the pathogen. The disad-
vantage of broad-spectrum antimicrobials is that by affecting a
Selective Toxicity wide range of microbes they disrupt the normal microbiota that
Medically useful antimicrobial drugs exhibit selective toxicity, play an important role in excluding pathogens. This in turn can
causing greater harm to microbes than to the human host. They do leave the patient predisposed to other infections.
460 Chapter 20 Antimicrobial Medications

Narrow-spectrum antimicrobials affect a limited range of as streptomycin can damage kidneys, impair the sense of bal-
bacteria. Their use requires that the pathogen be identified and its ance, and even cause irreversible deafness. Patients taking
antimicrobial susceptibility tested, but they cause less disruption these drugs must be closely monitored because of the low ther-
to the normal microbiota. normal microbiota, pp. 337, 381 apeutic index. Some antimicrobials have such severe potential
side effects that they are reserved for only life-threatening
conditions. In rare cases, for example, chloramphenicol causes
Effects of Combinations the potentially lethal condition aplastic anemia, in which the
Combinations of antimicrobials are sometimes used to treat infec- body is unable to make white and red blood cells. For this
tions, but these must be chosen carefully because some drugs reason, chloramphenicol is usually used only when no other
counteract the effects of others. Bacteriostatic drugs, for example, alternatives are available.
interfere with the action of drugs that kill only actively dividing
cells. Counteracting combinations such as this are antagonistic. In Suppression of the Normal Microbiota
contrast, combinations in which the activity of one drug enhances The normal microbiota plays an important role in host defense
the activity of the other are synergistic. Combinations that are by excluding pathogens. When the composition of the normal
neither synergistic nor antagonistic are additive. microbiota is altered, which happens when a person takes an anti-
microbial, pathogens normally unable to compete may multiply
to high numbers. Patients who take broad-spectrum antibiotics
Tissue Distribution, Metabolism, orally sometimes develop life-threatening antibiotic-associated
and Excretion of the Drug colitis, caused by toxin-producing strains of Clostridium difficile.
Antimicrobials differ not only in their action and activity, but This bacterium generally cannot establish itself in the intes-
also in how they are distributed, metabolized, and excreted tine due to competition from other bacteria. When members of
by the body. Only some drugs cross from the blood into the the normal intestinal microbiota are inhibited or killed, how-
cerebrospinal fluid, an important factor in treating meningitis. ever, C. difficile can sometimes flourish and cause serious intes-
Drugs that are unstable at low pH are destroyed by stomach acid tinal damage, resulting in a range of symptoms referred to as
when swallowed, so these drugs must instead be administered Clostridium difficile–associated disease. normal microbiota,
through intravenous or intramuscular injection. meningitis, p. 643 pp. 337, 381 Clostridium difficile–associated disease, p. 594
Another important characteristic of an antimicrobial drug is
its rate of elimination, expressed as the half-life. The half-life of
a drug is the time it takes for the serum concentration to decrease Resistance to Antimicrobials
by 50%. This dictates the frequency of doses required to maintain As pharmaceutical companies are assembling a vast array of
an effective level in the body. Penicillin V, which has a very short antimicrobial drugs, microorganisms are countering the efforts
half-life, needs to be taken four times a day, whereas azithromy- with a toolbox of mechanisms to avoid the drugs’ effects. Certain
cin, which has a half-life of over 24 hours, is taken only once a bacteria are inherently resistant to the effects of some drugs, a trait
day or less. Patients who have kidney or liver dysfunction often called intrinsic (innate) resistance. As an example, Mycoplasma
excrete or metabolize drugs more slowly, and so their drug dos- species lack a cell wall, so they are resistant to penicillin and any
ages must be adjusted accordingly to avoid toxic levels. other drug that interferes with peptidoglycan synthesis. Many
Gram-negative bacteria are intrinsically resistant to certain drugs
because the lipid bilayer of their outer membrane prevents the
Adverse Effects drug from entering (see figure 3.33). In other cases, previously
As with any medication, several concerns and dangers are associ- sensitive organisms develop acquired resistance through spon-
ated with antimicrobial drugs. It is important to remember, how- taneous mutation or horizontal gene transfer. The mechanisms
ever, that antimicrobials are extremely valuable drugs that save involved will be discussed later. the genus Mycoplasma, p.  276
countless lives when properly prescribed and used. Gram-negative cell wall, p. 60

Allergic Reactions
Some people develop allergies to antimicrobial drugs. An allergy MicroAssessment 20.2
to penicillin or related drug usually results in a fever or rash but When choosing an antimicrobial to prescribe, a variety of factors must
can abruptly cause life-threatening anaphylactic shock. For this be considered including the therapeutic index, antimicrobial action,
reason, people who have allergic reactions to a given antimicrobial spectrum of activity, effects of combinations, tissue distribution, half-
life, adverse effects, and resistance of the microbe.
must alert their physicians and pharmacists so an alternative drug
can be prescribed. They should also wear a bracelet or necklace 4. Which would be safest: an antimicrobial that has a low
that records that information in case of emergency. anaphylactic therapeutic index or one that has a high therapeutic index?
Why?
shock, p. 404
5. In what clinical situation is it most appropriate to use a broad-
spectrum antimicrobial?
Toxic Effects
6. Why would antimicrobials that have toxic effects be used
Several antimicrobials are toxic at high concentrations or at all? +
occasionally cause adverse reactions. Aminoglycosides such
 Part III Microorganisms and Humans 461
Focus Figure
Cell wall Nucleic acid synthesis
20.3 ■ Mechanisms of Action
(peptidoglycan)
synthesis
Fluoroquinolones
Rifamycins
of Antibacterial Drugs
β-lactam drugs
Vancomycin Learning Outcomes
Bacitracin 4. Describe the β-lactam drugs and other antimicrobials that
inhibit cell wall synthesis.
5. Describe the antimicrobial drugs that inhibit protein
synthesis.
6. Describe the antimicrobial drugs that inhibit nucleic acid
synthesis.
A B
7. Describe the antimicrobial drugs that inhibit metabolic
pathways.
8. Describe the antimicrobial drugs that interfere with cell
membrane integrity.
9. Describe the antibacterial medications used to treat
Cell membrane Metabolic pathways Protein synthesis Mycobacterium tuberculosis infections.
integrity (folate biosynthesis) Aminoglycosides
Polymyxin B Sulfonamides Tetracyclines This section describes the mechanisms of action of various
Daptomycin Trimethoprim Macrolides
Chloramphenicol antibacterial drugs, highlighting the bacterial processes and
Lincosamides structures targeted (figure  20.2). A group of antibiotics
Oxazolidinones called β-lactam drugs will be covered in the greatest detail,
Streptogramins
because they serve as excellent examples of some of the
FIGURE 20.2 Targets of Antibacterial Medications important features of antimicrobials in general. Table 20.1
? Why would Mycoplasma pneumoniae be intrinsically resistant to summarizes the antimicrobials discussed in this section.
β-lactam drugs?

TABLE 20.1 Characteristics of Antibacterial Drugs

Target/Drug Comments/Characteristics

Cell Wall Synthesis

β-lactam drugs Bactericidal against a variety of bacteria; inhibit penicillin-binding proteins. Resistance is due to synthesis of
β-lactamases, decreased affinity of penicillin-binding proteins, or decreased uptake.
Penicillins A family of antibacterial medications; different groups vary in their spectrum of activity and their
susceptibility to β-lactamases.
Natural penicillins: Active against Gram-positive and a few Gram-negative bacteria. Penicillin G is destroyed by stomach acid,
penicillin G, penicillin V and so it usually must be administered by injection. Penicillin V can be taken orally.
Penicillinase-resistant: Similar to natural penicillins, but resistant to inactivation by the penicillinase of staphylococci.
methicillin, dicloxacillin
Broad-spectrum: ampicillin, Similar to the natural penicillins, but more active against Gram-negative organisms.
amoxicillin
Extended-spectrum: Increased activity against Gram-negative rods, including Pseudomonas species.
ticarcillin, piperacillin
Cephalosporins A family of antibacterial medications. The later generations are generally more effective against Gram-
Cephalexin, cephradine, negative bacteria and less susceptible to destruction by β-lactamases but susceptible to extended-spectrum
cefaclor, cefprozil, cefixime, β-lactamases.
ceftibuten, cefepime
Carbapenems Resistant to inactivation by β-lactamases, but susceptible to carbapenemases. Imipenem must be given in
Imipenem, meropenem, combination with a drug that inhibits certain kidney enzymes in order to avoid its inactivation.
ertapenem, and doripenem
Monobactams Resistant to β-lactamases but susceptible to extended-spectrum β-lactamases; can be given to patients who
Aztreonam are allergic to penicillin. Primarily active against members of the family Enterobacteriaceae.
Vancomycin Bactericidal against Gram-positive bacteria; binds to the peptide side chain of N-acetylmuramic acid. Used to
treat serious systemic infections and severe Clostridium difficile–associated disease. In enterococci, resistance
is due to a plasmid-encoded altered target.
Bacitracin Bactericidal against Gram-positive bacteria; interferes with the transport of peptidoglycan precursors.
Common ingredient in non-prescription antibiotic ointments.

(continued)
462 Chapter 20 Antimicrobial Medications

TABLE 20.1 Characteristics of Antibacterial Drugs (Continued )

Target/Drug Comments/Characteristics
Protein Synthesis
Aminoglycosides Bactericidal against aerobic and facultative bacteria; bind to the 30S ribosomal subunit, blocking the
Streptomycin, gentamicin, initiation of translation and causing the misreading of mRNA. Toxicity limits the use. Resistance is due to
tobramycin, amikacin, a plasmid-encoded inactivating enzyme, alteration of the target molecule, or decreased uptake by a cell.
neomycin Neomycin is commonly used in non-prescription topical antibiotic ointments.
Tetracyclines Bacteriostatic against some Gram-positive and Gram-negative bacteria; bind to the 30S ribosomal subunit,
Tetracycline, doxycycline, blocking the attachment of tRNA. Resistance is generally due to decreased accumulation, either through
glycylcyclines decreased uptake or increased efflux.
Glycylcyclines Bacteriostatic against many Gram-positive and Gram-negative bacteria; bind to the 30S ribosomal subunit,
Tigecycline blocking the attachment of tRNA.
Macrolides Bacteriostatic against many Gram-positive bacteria as well as the most common causes of atypical
Erythromycin, clarithromycin, pneumonia; bind to the 50S ribosomal subunit, preventing the continuation of protein synthesis. Used for
azithromycin treating patients who are allergic to β-lactam drugs. Resistance is due to an inactivating enzyme, alteration
of the target molecule, or decreased uptake by a cell.
Chloramphenicol Bacteriostatic and broad-spectrum; binds to the 50S ribosomal subunit, preventing peptide bonds from
being formed. Generally used only as a last resort for life-threatening infections. Resistance is often due to
a plasmid-encoded inactivating enzyme.
Lincosamides Bacteriostatic against a variety of Gram-positive and Gram-negative bacteria, including the anaerobe
Lincomycin, clindamycin Bacteroides fragilis. Bind to the 50S ribosomal subunit, preventing the continuation of protein synthesis.
Associated with an even greater risk of developing Clostridium difficile–associated disease.
Oxazolidinones Bacteriostatic against a variety of Gram-positive bacteria. Bind to the 50S ribosomal subunit, interfering
Linezolid with the initiation of protein synthesis.
Streptogramins A synergistic combination of two drugs that bind to two different sites on the 50S ribosomal subunit,
Quinupristin, dalfopristin inhibiting distinct steps of protein synthesis. Individually each drug is bacteriostatic, but together they are
bactericidal. Effective against a variety of Gram-positive bacteria.
Nucleic Acid Synthesis
Fluoroquinolones Bactericidal against a wide variety of Gram-positive and Gram-negative bacteria; inhibit topoisomerases.
Ciprofloxacin, moxifloxacin Resistance is most often due to structural alterations in the topoisomerase target.
Rifamycins Bactericidal against Gram-positive and some Gram-negative bacteria. Bind RNA polymerase, blocking the
Rifampin initiation of RNA synthesis. Primarily used to treat infections caused by Mycobacterium tuberculosis and as
prophylaxis for patients who have been exposed to Neisseria meningitidis.
Metronidazole Bactericidal against anaerobes. Activated by anaerobic metabolism and then binds DNA, interfering
with synthesis and causing damaging breaks. Used to treat bacterial vaginosis and Clostridium difficile–
associated disease.
Folate Biosynthesis
Sulfonamides Bacteriostatic against a variety of Gram-positive and Gram-negative bacteria. Structurally similar to para-
aminobenzoic acid (PABA) and therefore inhibit the enzyme for which PABA is a substrate. Resistance is
most commonly due to a plasmid-encoded alternative enzyme.
Trimethoprim Often used in combination with a sulfa drug for a synergistic effect; inhibits the enzyme that catalyzes a
step following the one inhibited by the sulfonamides. Resistance is commonly due to a plasmid-encoded
alternative enzyme; the genes that encode resistance to sulfa drugs are often carried on the same plasmid.
Cell Membrane Integrity
Daptomycin Bactericidal against Gram-positive bacteria by damaging the cytoplasmic membrane.
Polymyxin B Bactericidal against Gram-negative bacteria by damaging cell membranes. Toxicity limits its use primarily to
topical applications, but it is a common ingredient in non-prescription antibiotic ointments.
Mycobacterium tuberculosis
Ethambutol Inhibits the synthesis of a component of the mycobacterial cell wall.
Isoniazid Inhibits synthesis of mycolic acid, a major component of the mycobacterial cell wall.
Pyrazinamide Mechanism unknown.
 Part III Microorganisms and Humans 463

Antibacterial Medications Penicillin

That Inhibit Cell Wall Synthesis O
S CH3
Bacterial cell walls are unique in that they contain peptidoglycan R C NH CH CH C
(see figures 3.32 and 3.33). Because of this, antimicrobial medi- CH3
cations that interfere solely with synthesis of this cell wall com-
ponent do not affect eukaryotic cells, and often have a very high
C N CH COOH
therapeutic index (figure 20.3).
O
Penicillins, Cephalosporins, β-lactam ring
and Other β-Lactam Drugs (a)
Penicillins and cephalosporins are members of a group of antimi-
crobial medications referred to as β-lactam drugs. This group, Cephalosporin
which also includes the monobactams and carbapenems, all have O S
a shared chemical structure called a β-lactam ring (figure 20.4).
The β-lactam drugs competitively inhibit a group of enzymes R C NH CH CH CH2
that catalyze the formation of peptide bridges between adjacent
glycan strands, an essential step in the final stages of peptido-
glycan synthesis (see figure 3.31). These enzymes are commonly C N C
called penicillin-binding proteins (PBPs), reflecting the fact
O R

β-lactam ring COOH

(b)
β-lactam drugs Vancomycin
Competitively inhibit enzymes Binds to the amino acid side FIGURE 20.4 The β-Lactam Ring of Penicillins and
that help form peptide bridges chain of NAM molecules, Cephalosporins The core chemical structure of (a) a penicillin; (b) a
between adjacent glycan blocking peptidoglycan cephalosporin. The β-lactam rings are marked by an orange circle. The
chains. synthesis. R groups vary among different penicillins and cephalosporins.
? Why is it not surprising that penicillin and cephalosporin both
have a high therapeutic index?

that they bind penicillin and were initially discovered during

experiments to study the effects of the medication. The disruption
Peptido-
glycan
in cell wall synthesis weakens the wall, causing the cell to lyse
(cell wall) (figure  20.5). The β-lactam drugs are bactericidal only against
growing bacteria, because these cells continuously synthesize
peptidoglycan. competitive enzyme inhibition, p. 138

Cytoplasmic
membrane

NAG

NAM

Bacitracin
Interferes with the transport
of peptidoglycan precursors
across the cytoplasmic
membrane.
FIGURE 20.5 Effect of a β-Lactam Drug on a Cell The drug
disrupts cell wall synthesis, leading to weakening of the cell wall,
FIGURE 20.3 Antibacterial Medications That Interfere with ultimately causing the cells to lyse.
Cell Wall Synthesis
? Why would a bacteriostatic drug be an antagonistic combination
? What is the function of penicillin-binding proteins? with penicillin?
464 Chapter 20 Antimicrobial Medications

The different β-lactam drugs vary in their spectrum of activ-

Side Chain Basic Structure
ity. One reason for this difference is the cell wall structure of
the bacteria. The peptidoglycan of Gram-positive organisms is
exposed to the outside environment, so the β-lactam drugs can O S CH3
directly contact the enzymes that synthesize the molecule. In con- CH2 C NH CH CH C
trast, the outer membrane of Gram-negative bacteria prevents the CH3
drugs from accessing their target. Another difference is the type of C N CH COOH
O
PBPs. The PBPs of Gram-positive bacteria differ somewhat from β-lactam ring
those of Gram-negative bacteria, and the PBPs of obligate anaer- Penicillin G
obes differ from those of aerobes. The various PBPs have different
affinities for the β-lactam drugs. Differences in affinity can even OCH2 Penicillin V
exist among related organisms such as Gram-positive cocci. (acid-resistant)
Some bacteria resist the effects of certain β-lactam drugs by
synthesizing a β-lactamase, an enzyme that breaks the critical
β-lactam ring, destroying the activity of the antibiotic. Just as OCH3
there are many β-lactam drugs, there are various β-lactamases, and
these differ in the range of drugs they destroy. Penicillinase is a Methicillin
(penicillinase-resistant)
β-lactamase that inactivates only members of the penicillin family.
In contrast, extended-spectrum β-lactamases (ESBLs) inactivate a OCH3
wide variety of β-lactam drugs, including both the penicillins and
the cephalosporins. As a whole, Gram-negative bacteria produce Cl
a much more extensive array of β-lactamases than Gram-positive
Dicloxacillin
organisms can. (acid- and penicillinase-resistant)
N
Cl O
The Penicillins All members of the penicillin family share a com- CH3
mon basic structure. This structure’s side chain has been chemi-
cally modified to create penicillin derivatives, each with unique
characteristics (figure  20.6). Penicillins can be loosely grouped
CH Ampicillin
into several categories: (broad-spectrum and acid-resistant)
NH2
■ Natural penicillins. These are the original penicillins pro-
duced naturally by the mold Penicillium chrysogenum.
Natural penicillins are narrow-spectrum antibiotics, effective
against Gram-positive and a few Gram-negative bacteria. HO CH Amoxicillin
(like ampicillin but more active
Penicillin V is more stable in acid and, therefore, better NH2 and requiring less frequent doses)
absorbed than penicillin G when taken orally. Bacteria that
produce penicillinase are resistant to the natural penicillins.
■ Penicillinase-resistant penicillins. Scientists developed CH Ticarcillin
these in a response to the problem of penicillinase-producing (more activity against Gram-negative
S COONa rods, including Pseudomonas, but not
Staphylococcus aureus strains. Penicillinase-resistant penicil- as effective against some Gram-positive
lins include methicillin and dicloxacillin. Unfortunately, some organisms)
penicillinase-producing bacteria acquired the ability to make
altered PBPs to which β-lactam drugs no longer bind. S. aureus
strains that can do this are called MRSA (methicillin-resistant CH Piperacillin
(like ticarcillin but a broader
S. aureus). NH spectrum of activity)
■ Broad-spectrum penicillins. The modified side chains of C O
these drugs give them a broad spectrum of activity. They N O
retain their activity against penicillin-sensitive Gram-positive
bacteria, yet they are also active against Gram-negative N O
bacteria. Unfortunately, they can be inactivated by many
C2H5
β-lactamases. Broad-spectrum penicillins include ampicillin
and amoxicillin.
FIGURE 20.6 Chemical Structures and Properties of
■ Extended-spectrum penicillins. These have greater activity Representative Members of the Penicillin Family The entire
against Pseudomonas species—Gram-negative bacteria that structure of penicillin G and the side chains of other penicillins are
are unaffected by many antimicrobial drugs. However, this shown.
group of penicillins has less activity against Gram-positive ? Could penicillin be used to treat MRSA (methicillin-resistant
bacteria. Like the other broad-spectrum penicillins, they are Staphylococcus aureus) infections? Why or why not?
 Part III Microorganisms and Humans 465

destroyed by many β-lactamases. Extended-spectrum penicil- Vancomycin does not cross the outer membrane of Gram-
lins include ticarcillin and piperacillin. negative bacteria, so these organisms are intrinsically resistant.
■ Penicillins + β-lactamase inhibitor. Rather than a new drug, Acquired resistance to vancomycin—an increasing problem—is
this is a combination of agents. The β-lactamase inhibitor most often due to a change in the peptide side chain of the NAM
interferes with the activity of some types of β-lactamases, molecule that prevents vancomycin from binding.
thereby protecting the penicillin against enzymatic destruc-
tion. An example is Augmentin, a combination of amoxicillin Bacitracin
and clavulanic acid. Bacitracin inhibits cell wall biosynthesis by interfering with the
transport of peptidoglycan precursors across the cytoplasmic
The Cephalosporins These are derived from an antibiotic pro- membrane. Its toxicity limits its use to topical applications (used
duced by the fungus Acremonium cephalosporium. Generally only on the surface of the skin); however, it is a common ingredi-
included in the cephalosporin family is a closely related group of ent in non-prescription first-aid ointments.
antibiotics made by filamentous bacteria related to Streptomyces.
The chemical structure of the cephalosporins makes them
resistant to inactivation by certain β-lactamases. Some are not Antibacterial Medications
very effective against Gram-positive bacteria, however, because That Inhibit Protein Synthesis
the drugs have a low affinity for the PBPs of these organisms. Several types of antibacterial drugs inhibit prokaryotic protein
The cephalosporins have been chemically modified, giving synthesis (figure  20.7). Although all cells synthesize proteins,
rise to first-, second-, third-, and fourth-generation versions. These the structure of the prokaryotic 70S ribosome—composed of a
include cephalexin and cephradine (first generation), cefaclor and 30S and a 50S subunit—is different enough from the eukaryotic
cefprozil (second generation), cefixime and ceftibuten (third gen- 80S ribosome to make it a suitable target for selective toxicity.
eration), and cefepime (fourth generation). The later generations The mitochondria of eukaryotic cells also have 70S ribosomes,
are generally more effective against Gram-negative bacteria and however, which may partially account for the toxicity of some of
less susceptible to destruction by β-lactamases. these drugs. ribosome structure, p. 66

Other β-Lactam Antibiotics Two other groups of β-lactam

The Aminoglycosides
drugs, carbapenems and monobactams, are resistant to most
β-lactamases. The carbapenems are effective against a wide range The aminoglycosides are bactericidal drugs that irreversibly bind
of Gram-negative and Gram-positive bacteria. Four types are to the 30S ribosomal subunit, causing it to distort and malfunction.
available—imipenem, meropenem, ertapenem, and doripenem. This blocks the initiation of translation and causes misreading of
Imipenem is rapidly destroyed by a kidney enzyme and is there- mRNA by ribosomes that have already passed the initiation step.
fore given in combination with a drug that inhibits that enzyme. Aminoglycosides are generally not effective against anaer-
Bacteria that produce carbapenemases are resistant. The only obes, enterococci, and streptococci. This is because they enter
monobactam used therapeutically, aztreonam, is primarily effec- bacterial cells by an active transport process that requires res-
tive against members of the family Enterobacteriaceae, which are piratory metabolism. To extend their spectrum of activity, the
Gram-negative rods. Aztreonam has a slightly different structure aminoglycosides are sometimes used in a synergistic combination
than other β-lactam drugs, so it can be given to patients who are
allergic to penicillin. Bacteria that produce ESBLs are resistant to
Macrolides
aztreonam. Enterobacteriaceae, p. 265 Prevent the continuation
Streptogramins
of protein synthesis.
Each interferes with a
Vancomycin distinct step of protein Chloramphenicol
Vancomycin blocks peptidoglycan synthesis by binding to the synthesis. Prevents peptide
bonds from being
peptide side chain of NAM molecules being assembled to form Lincosamides 50S formed.
glycan chains. This weakens the cell wall, causing cell lysis. The Prevent the
continuation of Oxazolidinones
drug is poorly absorbed from the intestinal tract, so it must be Interfere with the
protein synthesis.
administered intravenously except when used to treat intestinal initiation of protein
infections. NAM, p. 58
30S synthesis.
Vancomycin is a very important medication for treating
infections caused by Gram-positive bacteria resistant to β-lactam Tetracyclines and
Aminoglycosides
glycylcyclines
drugs. It is also used to treat severe cases of Clostridium Block the attachment
Block the initiation of
difficile–associated disease that do not respond to metronidazole. translation and cause the
of tRNA to the ribosome.
misreading of mRNA.
In fact, vancomycin is sometimes referred to as the antibiotic of last
resort because it is generally reserved for organisms resistant to all FIGURE 20.7 Antibacterial Medications That Inhibit
other options. Fortunately, new alternatives have been developed, Prokaryotic Protein Synthesis These medications bind to the
but vancomycin is still the drug of choice for treating most infec- 70S ribosome.
tions caused by Gram-positive bacteria resistant to other drugs. ? Some drugs that inhibit prokaryotic protein synthesis have a low
metronidazole, p. 478 therapeutic index. Why would this be so?
466 Chapter 20 Antimicrobial Medications

with a β-lactam drug. The β-lactam drug interferes with cell wall Chloramphenicol
synthesis, which, in turn, allows the aminoglycoside to enter cells Chloramphenicol binds to the 50S ribosomal subunit, preventing
that would otherwise be resistant. active transport, p. 56 peptide bonds from being formed and, consequently, blocking
Examples of aminoglycosides include streptomycin, gen- translation. Its action is bacteriostatic.
tamicin, tobramycin, and amikacin. Unfortunately, all of these Chloramphenicol is active against a wide range of bacteria,
can cause severe side effects including hearing loss and kid- but it is generally only used as a last resort for life-threatening
ney damage; consequently, they are generally used only when infections in order to avoid a rare but lethal side effect. This com-
other alternatives are not available. A form of tobramycin that plication, aplastic anemia, can occur in response to even a small
can be administered through inhalation rather than injection amount of chloramphenicol and is characterized by the inability of
makes treatment of lung infections in cystic fibrosis patients the body to form white and red blood cells.
caused by Pseudomonas aeruginosa safer and more effective.
Another aminoglycoside, neomycin, is too toxic for systemic The Lincosamides
use; however, it is a common ingredient in non-prescription The lincosamides bind to the 50S ribosomal subunit and prevent
topical ointments. the continuation of translation. Their action is bacteriostatic.
Lincosamides inhibit a variety of Gram-negative and Gram-
The Tetracyclines positive bacteria. They are particularly useful for treating infec-
The tetracyclines reversibly bind to the 30S ribosomal subunit, tions resulting from intestinal perforation because they inhibit
blocking the attachment of tRNA and preventing translation from Bacteroides fragilis, a member of the normal intestinal microbiota
continuing. These bacteriostatic drugs are actively transported into that is frequently resistant to other antimicrobials. Unfortunately,
prokaryotic but not animal cells, which effectively concentrates the risk of developing Clostridium difficile–associated disease is
them inside bacteria. This, in part, accounts for their selective greater for people taking lincosamides than some other antimicro-
toxicity. bials because most C. difficile strains are resistant to the lincos-
The tetracyclines are effective against certain Gram-positive amides. The most commonly used lincosamide is clindamycin.
and Gram-negative bacteria. Derivatives such as doxycycline have
a longer half-life, allowing less-frequent doses. Resistance to the The Oxazolidinones
tetracyclines is primarily due to a decrease in their accumula- The oxazolidinones bind to the 50S ribosomal subunit, interfering
tion by the bacterial cell, either by decreased uptake or increased with the initiation of translation. They are bacteriostatic against a
excretion. variety of Gram-positive bacteria and are useful in treating infec-
MicroByte tions caused by bacteria that are resistant to β-lactam drugs and
Tetracyclines can cause discoloration in teeth when used by young vancomycin. Linezolid is an example.
children.
The Streptogramins
Two streptogramins (quinupristin and dalfopristin) are admin-
The Glycylcyclines
istered together in a medication called Synercid. These act as
The glycylcyclines are a new class of antibacterial drugs related a synergistic combination, binding to two different sites on the
to the tetracyclines. They have the same mechanism of action 50S ribosomal subunit and inhibiting distinct steps of translation.
and effect as the tetracyclines, but a wider spectrum of activity. Individually, each drug is bacteriostatic but together they are bac-
In addition, they are effective against many bacteria that have tericidal. Synercid is effective against a variety of Gram-positive
acquired resistance to the tetracyclines. Tigecycline is the only bacteria, including some that are resistant to β-lactam drugs and
example currently approved. vancomycin.

The Macrolides
Antibacterial Medications That
The macrolides reversibly bind to the 50S ribosomal subunit and
prevent the continuation of translation. They often serve as the
Inhibit Nucleic Acid Synthesis
drug of choice for patients who are allergic to penicillin. Enzymes required for nucleic acid synthesis are the targets of
Macrolides are bacteriostatic against many Gram-positive some groups of antimicrobial drugs.
bacteria as well as the most common causes of atypical pneu-
monia (“walking pneumonia”). They are not effective against The Fluoroquinolones
members of the family Enterobacteriaceae because they do The fluoroquinolones are synthetic drugs that inhibit one or more
not pass through the outer membrane. Examples of macrolides of a group of enzymes called topoisomerases, which maintain the
include erythromycin, clarithromycin, and azithromycin. Both supercoiling of DNA within the bacterial cell. One type of topoi-
clarithromycin and azithromycin have a longer half-life than somerase, DNA gyrase, breaks and rejoins strands to relieve the
erythromycin, so that they can be taken less frequently. strain caused by the localized unwinding of DNA during replica-
Resistance can occur through modification of the ribosomal tion and transcription. Consequently, inhibition of this enzyme
RNA target, production of an enzyme that chemically modifies prevents these essential cell processes. supercoiled DNA, p. 66
the drug, and alterations that result in decreased drug uptake. The fluoroquinolones are bactericidal against a wide variety
walking pneumonia, p. 500 of bacteria, including both Gram-positive and Gram-negative
 Part III Microorganisms and Humans 467

organisms. Examples of fluoroquinolones include ciprofloxacin

and moxifloxacin. Acquired resistance is most commonly due to Para-aminobenzoic Precursor
acid (PABA) #1
an alteration in the DNA gyrase target.
Enzyme
PABA Sulfa drugs
The Rifamycins H 2N COOH #1

The rifamycins block prokaryotic RNA polymerase from initiat- Precursor

ing transcription. Rifampin, the most widely used rifamycin, is #2
bactericidal against many Gram-positive and some Gram-negative Sulfanilamide
Enzyme
Glutamate
bacteria as well as members of the genus Mycobacterium. #2
H2N SO2NH2
Rifampin is primarily used to treat tuberculosis and Hansen’s
Dihydrofolate
disease (leprosy) and to prevent meningitis in people who have
been exposed to Neisseria meningitidis. In some patients, a (a) Enzyme
Trimethoprim
reddish-orange pigment appears in urine and tears. Resistance to #3
the drug develops rapidly and is due to a mutation in the gene that FIGURE 20.8 Inhibitors
of the Folate Pathway Tetrahydrofolate
encodes RNA polymerase. (a) The chemical structure
of PABA and a sulfa drug Multiple enzymes
Metronidazole (sulfanilamide). (b) The sul- and reactions

Metronidazole (Flagyl) interferes with DNA synthesis and func- fonamides and trimethoprim Thymine, guanine,
interfere with different steps and adenine
tion, but only in anaerobic microorganisms. The selective toxicity of the pathway that leads ini- nucleotides
is due to the fact that anaerobic metabolism is required to convert tially to the synthesis of folate
the medication to its active form. The active form then binds and ultimately to the synthe- (b)
DNA, interfering with synthesis and causing damaging breaks. sis of a coenzyme required for
Metronidazole is used to treat bacterial vaginosis and Clostridium nucleotide biosynthesis.
difficile–associated disease. bacterial vaginosis, p. 617 ? Sulfa drugs have a high therapeutic index. Why would this be so?

Antibacterial Medications That of resistance is a plasmid-encoded alternative enzyme that has

Interfere with Metabolic Pathways a lower affinity for the drug. Unfortunately, the genes encoding
Relatively few antibacterial medications interfere with metabolic resistance to trimethoprim and sulfonamide are often carried on
pathways. Among the most useful are the folate inhibitors— the same plasmid.
sulfonamides and trimethoprim. These each inhibit different steps
in the pathway that leads initially to the synthesis of folate and
ultimately to the synthesis of a coenzyme required for nucleotide Antibacterial Medications That Interfere
biosynthesis (figure 20.8). Animal cells lack the enzymes in the with Cell Membrane Integrity
folate synthesis portion of the pathway, which is why folic acid is A few antimicrobial drugs damage bacterial membranes. They
a dietary requirement. coenzyme, p. 136 cause the cells to leak, leading to cell death.
Daptomycin inserts into bacterial cytoplasmic membranes,
The Sulfonamides and is used to treat certain infections caused by Gram-positive
Sulfonamides and related compounds, collectively referred to bacteria resistant to other drugs. It is not effective against Gram-
as sulfa drugs, inhibit the growth of many Gram-positive and negative bacteria because it cannot penetrate the outer membrane.
Gram-negative bacteria. They are structurally similar to para- Polymyxin B, a common ingredient in first-aid skin ointments,
aminobenzoic acid (PABA), a substrate in the pathway for folate binds to the membranes of Gram-negative cells. Unfortunately,
biosynthesis. Because of this similarity, the enzyme that normally these drugs also bind to eukaryotic cells, though to a lesser extent,
binds PABA binds sulfa drugs instead, an example of competi- which generally limits their use to topical applications.
tive inhibition (see figure  6.15). Human cells lack this enzyme,
providing the basis for the selective toxicity of the sulfonamides.
Resistance to the sulfonamides is often due to the acquisition of Antibacterial Medications Effective
a plasmid-encoded enzyme that has a lower affinity for the drug. Against Mycobacterium Species
competitive inhibition, p. 138 Relatively few antimicrobials are effective against Mycobacterium
tuberculosis and related species. This is due to several factors,
Trimethoprim including the organism’s waxy cell wall (which prevents the entry
Trimethoprim inhibits the bacterial enzyme that catalyzes a of many drugs) and slow growth. A group of five medications—the
metabolic step following the one inhibited by sulfonamides. first-line drugs—are preferred because they are the most effective
Fortunately, the drug has little effect on the enzyme’s counterpart as well as the least toxic. These are generally given in combination
in human cells. The combination of trimethoprim and a sulfon- of two or more to patients who have active tuberculosis disease.
amide has a synergistic effect, and they are often used together This combination therapy decreases the chance that resistant
to treat urinary tract infections. The most common mechanism mutants will develop; if some cells in the infecting population
468 Chapter 20 Antimicrobial Medications

spontaneously develop resistance to one drug, the other drug will strain can be successfully treated with the drug. For example, an
eliminate them. The second-line drugs are used for strains resis- organism with an MIC of 16 μg/ml would be considered resistant
tant to the first-line drugs; however, they either are less effective to a drug if the level that can be achieved in a person’s blood
or have greater risk of toxicity. Mycobacterium tuberculosis, p. 503 is less than that. Microbes that have an MIC on the borderline
Of the first-line medications, some specifically target the between susceptible (treatable) and resistant (untreatable) are
unique cell wall that characterizes the mycobacteria. Isoniazid called intermediate.
inhibits the synthesis of mycolic acids, a primary component of The minimum bactericidal concentration (MBC) is the
the cell wall. Ethambutol inhibits enzymes required for synthesis lowest concentration of a specific antimicrobial drug that
of other mycobacterial cell wall components. The mechanism of kills 99.9% of cells of a given bacterial strain in vitro. The
pyrazinamide is unknown. Other first-line drugs include rifampin MBC is determined by finding out how many live organisms
and streptomycin, which have already been discussed. remain in tubes from the MIC test that showed no growth.
A small sample from each of those tubes is transferred to a
MicroAssessment 20.3 plate containing an antibiotic-free agar medium. If a sample
The targets of antimicrobial drugs include biosynthetic pathways for
gives rise to no colonies, then no cells survived that particular
peptidoglycan, protein, nucleic acid, and folic acid and the integrity drug concentration, indicating it was bactericidal. If colonies
of membranes. Drugs used to treat tuberculosis often interfere with form, they can be counted to determine the number of cells
processes unique to Mycobacterium tuberculosis. that survived.
7. Why are β-lactam drugs only bactericidal to growing bacteria? Determining the MIC and MBC using these methods gives
8. What is the target of the macrolides?
precise information regarding an organism’s susceptibility. The
techniques, however, are labor-intensive and therefore expensive.
9. Considering that all β-lactam drugs have the same target, why
do they vary in their spectrum of activity? +
In addition, individual sets of tubes must be inoculated to deter-
mine susceptibility to each drug tested (see Perspective 20.1).

20.4 ■ Determining the Conventional Disc Diffusion Method

Susceptibility of a The Kirby-Bauer disc diffusion test is routinely used to deter-
Bacterial Strain to an mine the susceptibility of a given bacterial strain to a battery of
antimicrobial drugs. A standard concentration of the strain is first
Antimicrobial Drug uniformly spread on the surface of an agar plate. Then 12 or so
Learning Outcomes discs, each containing a known amount of a different drug, are
placed on the surface of the medium (figure 20.10). During incu-
10. Describe how the minimum inhibitory concentration (MIC) and
the minimum bactericidal concentration (MBC) are determined. bation, the various drugs diffuse outward from the discs, forming a
concentration gradient around each disc. Meanwhile the bacterial
11. Compare and contrast the Kirby-Bauer disc diffusion test with
commercial modifications of antimicrobial susceptibility testing. cells multiply, eventually forming a film of growth on the plate,
except in regions around the discs where the bacteria were killed
Susceptibility of a pathogen to a specific antimicrobial drug is or their growth inhibited.
often unpredictable. In these cases, laboratory tests are used to A clear zone of inhibition around an antimicrobial disc
determine the susceptibility of the organism to various drugs and reflects, in part, the degree of susceptibility of the organism to
then choose the one that acts against the pathogen but as few other the drug. The zone size is also influenced by characteristics of the
bacteria as possible. drug, including its molecular weight and stability, as well as the
amount in the disc.
Special charts have been prepared correlating the size of the
Minimum Inhibitory and Bactericidal zone of inhibition to susceptibility of bacteria to the drug. Based
Concentrations (MIC and MBC) on the size of the zone, organisms can be described as susceptible,
intermediate, or resistant to the drug.
The minimum inhibitory concentration (MIC) is the lowest
concentration of a specific antimicrobial drug needed to prevent
the growth of a given bacterial strain in vitro. This is determined
by growing the test strain in broth cultures containing different
Commercial Modifications
concentrations of the antimicrobial (figure 20.9). To do this, serial of Antimicrobial Susceptibility Testing
dilutions are used to generate decreasing concentrations of the Commercial modifications of the conventional methods offer cer-
drug in tubes containing a suitable growth medium. Then, a fixed tain advantages. They are less labor-intensive, and the results can
concentration of bacterial cells is added to each tube. The tubes be obtained in as little as 4 hours. One system uses a small card
are incubated for at least 16 hours and then examined for turbidity, with tiny wells containing specific antimicrobial concentrations.
which indicates growth. The lowest concentration of the drug that The highly automated system inoculates and incubates the cards,
prevents growth of the microorganism is the MIC. determines the growth rate by reading the turbidity, and uses
The fact that a strain is inhibited by a given concentration of mathematical formulas to interpret the results and determine the
drug does not necessarily mean that an infection caused by that MICs in 6 to 15 hours (figure 20.11).
 Decreasing Concentration of the Antimicrobial Drug

Organism A

Control 16 µg/ml 8.0 4.0 2.0 1.0 0.5 0.25 0.12 0.06 0.03 Control
(no bacteria) (no drug)
Result: MIC = 0.12 µg/ml

Organism B

Control 16 µg/ml 8.0 4.0 2.0 1.0 0.5 0.25 0.12 0.06 0.03 Control
(no bacteria) (no drug)
Result: MIC = 1.0 µg/ml

Organism C

Control 16 µg/ml 8.0 4.0 2.0 1.0 0.5 0.25 0.12 0.06 0.03 Control
(no bacteria) (no drug)
Result: MIC = 16 µg/ml

FIGURE 20.9 Determining the Minimum Inhibitory Concentration (MIC) of an Antimicrobial Drug The lowest concentration of drug
that prevents growth of the culture is the MIC.
? Considering that organism C is susceptible to 16 μg/ml of the drug, why might it still be considered resistant to the drug?

FIGURE 20.11 Automated Tests Used to Determine

FIGURE 20.10 Kirby-Bauer Method for Determining Drug Antimicrobial Susceptibility The miniature wells in the card con-
Susceptibility The size of the zone of inhibition surrounding the tain specific concentrations of an antimicrobial drug. An automated
disc reflects, in part, the sensitivity of the bacterial strain to the system inoculates and incubates the cards, determines the growth
drug. rate by reading turbidity, and uses mathematical formulas to inter-
pret the results and derive the MICs.
? When using the Kirby-Bauer test to determine drug susceptibility,
a chart must be consulted. Why not simply choose the drug that ? What are two advantages of automated tests used to determine
gives the largest zone size? antimicrobial susceptibility?
470 Chapter 20 Antimicrobial Medications

PERSPECTIVE 20.1
Measuring the Concentration of an Antimicrobial Drug in Blood or Other Body Fluids
Sometimes it is necessary to determine the

Concentration of drug (μg/ml) (logarithmic scale)

concentration of an antimicrobial drug in 6.25
a patient’s blood or other body fluid. For
example, patients who are being administered
an aminoglycoside must often be carefully C
monitored to ensure that the concentration
1.25
of the drug in their blood does not reach an
unsafe level, particularly if they have kidney
or liver dysfunction that interferes with normal
elimination. Likewise, newly developed drugs
must be tested to determine achievable levels 0.25
in the blood, urine, or other body fluids.
A diffusion bioassay is used to measure
the concentration of an antimicrobial drug in
a fluid specimen. The test relies on the same
principle as the Kirby-Bauer test, except in
10 20 D 30
this case it is the concentration of drug being
Zone diameter (mm)
determined, not the sensitivity of the organism.
To do a diffusion bioassay, a culture of
Standards and patient’s serum are A standard curve that correlates the zone diameter
a stock organism highly susceptible to the added to agar that has been seeded with the antimicrobial drug concentration is constructed.
drug in question is added to melted cooled with susceptible strain of bacteria. The drug concentration in the serum can be read
agar, and the mixture poured into an agar from the line relating zone size to concentration.
plate and allowed to solidify. This results in (a)
(b)
a solid medium uniformly inoculated with FIGURE 1 Diffusion Bioassay
the sensitive organism. Cylindrical holes are
then punched out of the agar, creating wells.
Standards (known concentrations of the drug) which correspond to the concentrations of the plotting the zone sizes against the correspond-
are then added to some of the wells, while drug—the higher the concentration, the larger ing drug concentration. A line relating zone
others are filled with the body fluid being the zone of inhibition (figure 1). The zone size to concentration is obtained, from which
tested. Zones of inhibition develop around the sizes around the standards are measured, and the concentration of the antimicrobial drug in
wells during overnight incubation, the sizes of from this a standard curve is constructed by the body fluid can be read.

The E test, a modification of the disc diffusion test, uses a

strip containing a gradient of concentrations of an antimicrobial
drug. Multiple strips, each one containing a different drug,
are placed on the surface of an agar medium that has been
uniformly inoculated with the test organism. During
incubation, the test organism will grow, and a zone of
inhibition will form around the strip. Because of the
gradient of drug concentrations, the zone of inhibi-
tion will be shaped somewhat like a teardrop that
intersects the strip at some point (figure 20.12).
The MIC is determined by reading the printed
number at the point where the bacterial growth
intersects the strip.

FIGURE 20.12 The E Test (a) Each strip has a gradi-

ent of concentrations of a different antimicrobial drug.
(b) The MIC is determined by reading the number on
the strip at the point at which growth intersects the
strip.
? Why is the zone of inhibition larger at one end of
the strip? (a) (b)
 Part III Microorganisms and Humans 471

MicroAssessment 20.4
S
The MIC and MBC are quantitative measures of a bacterial strain’s
susceptibility to an antimicrobial drug. Disc diffusion tests can
determine whether an organism is susceptible, intermediate, or S
resistant to a battery of different drugs. Commercial tests for S
S
determining antimicrobial sensitivity are less labor-intensive and
often more rapid.
R
10. Explain the difference between the MIC and the MBC.
11. List two factors other than a strain’s sensitivity that influence the S
S
size of the zone of inhibition around an antimicrobial disc.
12. Why would it be important for the Kirby-Bauer disc diffusion
Antimicrobial drug is added;
test to use a standard concentration of the bacterial strain being sensitive organisms are killed
tested? + or inhibited.

20.5 ■ Resistance to S
Antimicrobial Drugs S S

Learning Outcomes
R
12. Describe four general mechanisms of antimicrobial
resistance. S
S
13. Describe how antimicrobial resistance can be acquired.
14. List five examples of emerging antimicrobial resistance.
Resistant survivors can
15. Describe how the emergence and spread of antimicrobial multiply without competition.
resistance can be slowed.

After sulfa drugs and penicillin were introduced, people hoped

that such drugs would eliminate most bacterial diseases. We now R
recognize, however, that drug resistance limits the usefulness of
all known antimicrobials.
As antimicrobial drugs are increasingly used and misused,
R R
resistant bacterial strains have a selective advantage over their
sensitive counterparts (figure 20.13). For example, when penicil-
lin G was first introduced, less than 3% of Staphylococcus aureus
strains were resistant to its effects. Heavy use of the drug, mea-
R R R R
sured in hundreds of tons per year, eliminated sensitive strains, so
that 90% or more are now resistant.
Antimicrobial resistance is alarming because of the impact
on the cost, complications, and outcomes of treatment. Dealing FIGURE 20.13 The Selective Advantage of Drug Resistance
with the problem requires an understanding of the mechanisms of When antimicrobial drugs are used, bacterial strains that are resistant
resistance and how they are spread. A better understanding of drug (R) to their effects have a selective advantage over their sensitive (S)
resistance may also allow scientists to develop new drugs that foil counterparts.
common resistance mechanisms. ? How does overuse of antibiotics contribute to increasing numbers
of drug-resistant bacteria?

Mechanisms of Acquired Resistance

Figure 20.14 depicts the most common mechanisms of acquired Alteration in the Target Molecule
resistance to antimicrobial drugs.
Antimicrobial drugs generally act by recognizing and bind-
ing to specific target molecules in a bacterium, interfering
Drug-Inactivating Enzymes with their function. Minor structural changes in the target can
Some bacteria produce enzymes that chemically modify a specific prevent the drug from binding. Modifications in the penicillin-
drug, interfering with its function. One example is penicillinase, binding proteins (PBPs) prevent β-lactam drugs from binding
which destroys penicillin. Another is the enzyme chloramphenicol to them. Similarly, a change in ribosomal RNA, the target for
acetyltransferase, which chemically alters the antibiotic chloram- the macrolides, prevents those drugs from interfering with
phenicol, making it ineffective. ribosome function.
472 Chapter 20 Antimicrobial Medications

Spontaneous Mutation
Non-resistant cell
As cells replicate, spontaneous mutations happen at a relatively
low rate. Those few mutations that occur, however, can have a
significant effect on the resistance of a bacterial population to an
Target
antimicrobial drug.
Drug
Drug binds Acquired resistance to streptomycin (an aminoglycoside) is
target. a good example of the consequence of spontaneous mutation. A
single base-pair change in the gene encoding a ribosomal protein
alters the target enough to make the cell streptomycin-resistant.
When a streptomycin-sensitive strain is grown in streptomycin-
free medium to a population of 109 cells, at least one cell in the
population probably has that particular mutation. If streptomycin
is then added to the medium, only that cell and its progeny will
Resistant cell be able to replicate, giving rise to a streptomycin-resistant popula-
tion. spontaneous mutation, p. 190
Increased elimination Drug-inactivating enzyme When an antimicrobial drug has several different targets or
Drug enters cell but Enzyme modifies has multiple binding sites on a single target, resistance through
efflux pump ejects it. drug, inactivating it.
spontaneous mutation is less likely to occur. This is because
several different mutations are required to prevent binding of
the drug. The newer aminoglycosides bind to several sites on the
ribosome, so resistance due to spontaneous mutation is unlikely.
Drugs such as streptomycin for which a single point mutation
causes resistance are sometimes used in combination with one or
more other drugs. If any cell spontaneously develops resistance to
Alteration in one drug, another drug will still kill it. Combination therapy is
target effective because the chance of a cell simultaneously developing
Decreased uptake molecule
Porin proteins prevent Drug cannot mutational resistance to multiple drugs is extremely low.
entry into the cell. bind target.
Gene Transfer
FIGURE 20.14 Common Mechanisms of Acquired Genes encoding resistance to antimicrobial drugs can spread
Antimicrobial Drug Resistance to different strains, species, and even genera, most commonly
? Strains of MRSA (methicillin-resistant Staphylococcus aureus) use through conjugative transfer of R plasmids. These plasmids often
which two methods to avoid the bactericidal effects of β-lactam carry several different resistance genes, each one encoding resis-
drugs? tance to a specific antimicrobial drug. Thus, when an organism
acquires an R plasmid, it becomes resistant to several different
Decreased Uptake of the Drug medications simultaneously. R plasmid, p. 209 conjugation, p. 206

The porin proteins in the outer membrane of Gram-negative bacte- In some cases, resistance genes originate through spontane-
ria selectively permit small hydrophobic molecules to enter a cell. ous mutation of common bacterial genes, such as one encoding the
Changes in these proteins can therefore prevent certain drugs from target of the drug. In other cases, the genes may have originated
entering the cell. By stopping entry of a drug, an organism avoids from the soil microbes that naturally produce that antibiotic. For
its effects. porins, p. 60
example, a gene coding for an enzyme that chemically modifies an
aminoglycoside likely originated from the Streptomyces species
Increased Elimination of the Drug that produces the drug.
The systems that bacteria use to transport damaging compounds
out of a cell are called efflux pumps. When a cell makes Examples of Emerging
more of these pumps, it can eject the drug faster. In addition, Antimicrobial Resistance
structural changes in the pumps can influence the range of Some of the problems associated with the increasing antimicrobial
drugs that can be pumped out. Resistance that develops by this resistance are highlighted by the following examples.
mechanism is particularly worrisome because it might allow an
organism to become resistant to several drugs simultaneously. Enterococci
efflux pumps, p. 56 One of the most dramatic examples of antimicrobial resistance is
the enterococci, a group of bacteria that is part of the normal intes-
tinal microbiota and a common cause of healthcare-associated
Acquisition of Resistance infections. Enterococci are intrinsically less susceptible to many
Antimicrobial resistance can be due to either spontaneous muta- common antimicrobials. For example, their penicillin-binding
tion, which alters existing genes, or acquisition of new genes (see proteins have low affinity for certain β-lactam antibiotics. In
figure 8.1). addition, many enterococci have R plasmids. Some strains, called
 Part III Microorganisms and Humans 473

vancomycin-resistant enterococci (VRE), are even resistant to Many tuberculosis patients do not comply with the complex
vancomycin. Recall that this drug is usually reserved as a last course of combination therapy, skipping doses or stopping treat-
resort for treating life-threatening infections caused by Gram- ment too soon. As a consequence, strains of M. tuberculosis
positive organisms resistant to all β-lactam drugs. Because van- develop resistance to the first-line drugs. This results in even lon-
comycin resistance in these strains is encoded on a plasmid, the ger, more expensive treatments that are also less effective.
resistance is transferable to other organisms. healthcare-associated Strains resistant to two of the favored drugs for tuberculo-
infections, p. 449 sis treatment—isoniazid and rifampin—are called multidrug-
resistant M. tuberculosis (MDR-TB). To prevent the emergence
Staphylococcus aureus of these strains, some cities are using directly observed therapy;
Staphylococcus aureus, another common cause of healthcare- healthcare workers routinely visit patients and watch them take their
associated infections, is becoming increasingly resistant to anti- drugs to make sure they comply with their prescribed antimicrobial
microbials. Although nearly all strains were susceptible to penicillin treatment. Strains of extensively drug-resistant M. tuberculosis
when it was first introduced 70 years ago, most are now resistant due (XDR-TB) are an even greater concern. These are defined as
to their acquisition of a gene encoding penicillinase. Until recently, M. tuberculosis strains resistant to isoniazid and rifampin, plus three
infections by these strains could be treated with methicillin or other or more of the second-line drugs. directly observed therapy, p. 505
penicillinase-resistant penicillins. New strains have emerged, how-
ever, that not only produce penicillinase, but also have penicillin- Enterobacteriaceae
binding proteins with low affinity for all β-lactam drugs. These strains, Members of the family Enterobacteriaceae are intrinsically
called methicillin-resistant Staphylococcus aureus (MRSA), are resistant to many antimicrobial medications because their outer
resistant to methicillin as well as all other β-lactam drugs. membrane prevents the drugs from entering cells. The situation
There are two categories of MRSA strains—healthcare- became more complicated when some enterics developed the
associated (HA-MRSA) and community acquired (CA-MRSA). ability to produce a β-lactamase, allowing the strains to resist
HA-MRSA strains are generally resistant to a wide range of anti- the effects of ampicillin and other penicillins. Some strains then
microbial medications, so they are generally treated with vancomy- developed the ability to produce extended-spectrum β-lactamases
cin. A few hospitals, however, have reported isolates that are no (ESBLs), making them resistant to most cephalosporins and
longer susceptible to normal levels of vancomycin. So far, strict aztreonam, as well as penicillins. More recently, strains referred
hospital guidelines designed to immediately halt the spread of these to as carbapenem-resistant Enterobacteriaceae (CRE) have
vancomycin-intermediate S. aureus (VISA) and vancomycin- been discovered. These strains, which are resistant to nearly all
resistant S. aureus (VRSA) strains have been successful. available antimicrobial drugs, produce an enzyme that inactivates
Fortunately, most CA-MRSA strains are currently susceptible to carbapenems, a group of drugs considered a last resort for treating
antibiotics other than β-lactam drugs. Staphylococcus aureus, p. 524 infections caused by ESBL-producing bacteria.

Streptococcus pneumoniae
Until recently, Streptococcus pneumoniae, the leading cause Slowing the Emergence and Spread
of pneumonia in adults, has remained very sensitive to penicil- of Antimicrobial Resistance
lin. Some isolates, however, are now resistant to the drug. This To reverse the alarming trend of increasing antimicrobial drug
acquired resistance is due not to the production of a β-lactamase, resistance, everyone must cooperate. On an individual level, physi-
but rather to changes in the chromosomal genes coding for the tar- cians as well as the general public must take more responsibility for
gets of penicillin—the penicillin-binding proteins. The modified the appropriate use of these life-saving medications. On a global scale,
targets have lower affinities for the drug. The nucleotide changes countries around the world need to make important policy decisions
do not appear to have come from point mutations as one might about what is, and what is not, an appropriate use of these drugs.
expect; instead, they are due to the acquisition of chromosomal
DNA from other species of Streptococcus. As you may recall The Responsibilities of Physicians
from earlier reading, S. pneumoniae can acquire DNA through and Other Healthcare Workers
DNA-mediated transformation. Streptococcus pneumoniae, p.  497 Physicians and other healthcare workers need to increase their
DNA-mediated transformation, p. 202 efforts to identify the cause of a given infection and, only if appro-
priate, prescribe suitable antimicrobials. They must also educate
Mycobacterium tuberculosis patients about the proper use of prescribed drugs in order to increase
Tuberculosis treatment has always been a long and compli- compliance. Although these efforts may be more expensive in the
cated process, requiring a combination of two or more different short term, they will ultimately save both lives and money.
drugs taken for a period of 6 months or more. Unfortunately,
Mycobacterium tuberculosis can easily become resistant to the The Responsibilities of Patients
first-line drugs (the preferred medications) through spontaneous Patients need to carefully follow the instructions that accompany
mutation. Large numbers of bacterial cells are found in an active their prescriptions, even if those instructions seem inconvenient.
infection, so it is likely that at least one cell has developed spon- It is essential to maintain adequate blood levels of the antimicro-
taneous resistance to a drug, which is why combination therapy is bial for a specific time period. When a patient skips a scheduled
required. The length of treatment is due to the very slow growth dose of a drug, the blood level of the drug may not remain high
of M. tuberculosis. Mycobacterium tuberculosis, p. 503 enough to inhibit the growth of the least-sensitive members of the
474 Chapter 20 Antimicrobial Medications

population. If these less-sensitive organisms then have a chance 20.6 ■ Mechanisms of Action
to grow, they will give rise to a population that is not as sensi-
tive as the original. Likewise, failure to complete the prescribed of Antiviral Drugs
course of treatment may not kill the least-sensitive organisms,
Learning Outcomes
allowing their subsequent multiplication. Misusing antimicrobi-
als by skipping doses or failing to complete the prescribed treat- 16. Describe the antiviral drugs that prevent viral entry or uncoating.
ment increases the likelihood that resistant mutants will develop. 17. Compare and contrast the antiviral drugs that interfere with
nucleic acid synthesis or genome integration.
The Importance of an Educated Public 18. Describe the antiviral drugs that interfere with the assembly and
release of viral particles.
A greater effort must also be made to educate people about the
role and limitations of antibiotics in order to make sure they are
used wisely. First and foremost, people need to understand that Viruses rely almost exclusively on the host cell’s metabolic machin-
antibiotics are not effective against viruses. Taking antibiotics will ery for their replication, making it difficult to find a target for
not cure the common cold or any other viral illness. A few antivi- selective toxicity. They have no cell wall, ribosomes, or any other
ral drugs are available, but they are effective against only certain structure targeted by antibiotics. Because of this, viruses are com-
viruses. Unfortunately, surveys indicate that far too many people pletely unaffected by antibiotics. Many encode their own polymer-
mistakenly believe that antibiotics are effective against viruses, ases, however, and these are potential targets of antiviral drugs
and often seek prescriptions to “cure” viral infections. This misuse (figure 20.15). Relatively few other targets have been discovered.
only selects for antibiotic-resistant bacteria in the normal micro- Many scientists are trying to develop more effective antiviral
biota. Even though these organisms typically do not cause disease, drugs, medications that interfere with viral replication. The rela-
they can serve as a reservoir for R plasmids, eventually transfer- tively few options available are generally effective against only a
ring their resistance genes to an infecting pathogen. specific type of virus, and none can eliminate latent viral infec-
tions (table 20.2). latent viral infections, p. 322

Global Impacts of the Use of Antimicrobial Drugs

The overuse of antimicrobial drugs is a worldwide concern. Coun- Prevent Viral Entry
tries may vary in their laws and customs, but antimicrobial resistance A new group of drugs effective against HIV are the entry inhibi-
recognizes no political boundaries. An organism that develops resis- tors, which prevent the virus from entering host cells. Enfuvirtide
tance in one country can quickly be transported globally. does this by binding to an HIV protein that promotes fusion of the
In many parts of the world, particularly in developing coun- viral envelope with the cell membrane. Maraviroc blocks the HIV
tries, antimicrobial drugs are available on a non-prescription basis. co-receptor CCR5. HIV attachment and entry, p. 699
Because of the consequences of inappropriate use, many people
Entry inhibitors
believe that over-the-counter availability of these drugs should be Effective against HIV
restricted or eliminated.
Eukaryotic
Another worldwide concern is the use of antimicrobial drugs Entry
host cell
in animal feeds. Low levels of these drugs in feeds enhance the
growth of animals, a seemingly attractive option. This use, like Uncoating
any other, however, selects for drug-resistant organisms, which Nucleic acid synthesis
(viral enzyme directed)
has caused some scientists to question its ultimate wisdom. In Integrase
fact, infections caused by drug-resistant Salmonella strains have inhibitors
Viral particle
Effective
been linked to animals whose feed was supplemented with those production
against HIV
drugs. In response to these concerns, there is growing pressure
worldwide to ban the use of antimicrobial drugs in animal feeds.
Exit

MicroAssessment 20.5 Viral uncoating

Effective against
Mutations and transfer of genetic information allow microorganisms influenza A virus Assembly and release
to become resistant to antimicrobial medications. Drug resistance Amantadine of viral particles
affects the outcomes of medical treatment. Slowing the emergence Rimantadine Effective against HIV
and spread of resistant microbes involves the cooperation of Protease inhibitors
Nucleic acid synthesis Effective against influenza viruses
healthcare personnel, educators, and the general public. Effective against herpesviruses Neuraminidase inhibitors
13. Explain how using a combination of two antimicrobial drugs Nucleoside analogs
helps prevent the development of spontaneously resistant Non-nucleoside polymerase
inhibitors
mutants. Effective against HIV
14. Explain the significance of a member of the normal flora that Nucleoside analogs
harbors an R plasmid. Non-nucleoside reverse
transcriptase inhibitors
15. A student argued that “spontaneous mutation” meant that a
drug could cause mutations. Is the student correct? Why or FIGURE 20.15 Targets of Antiviral Drugs
why not? +
? Why are there relatively few options for antiviral drugs?
 Part III Microorganisms and Humans 475

TABLE 20.2 Characteristics of Antiviral Drugs

Target/Drug Examples Comments/Characteristics

Viral Entry
Enfuvirtide, Maraviroc Used to treat HIV infections.
Viral Uncoating
Amantadine and rimantadine Reduce severity and duration of influenza A infections, but resistance limits their use.
Nucleic Acid Synthesis
Nucleoside analogs Primarily used to treat infections caused by herpesviruses and HIV; they do not cure latent
Acyclovir, ganciclovir, ribavirin, zidovudine infections. The drugs are converted within eukaryotic cells to a nucleotide analog; virally
(AZT), didanosine (ddl), lamivudine (3TC) encoded enzymes are prone to incorporate these, resulting in premature termination of
synthesis or improper base-pairing of the viral nucleic acid. Acyclovir is used to treat herpes
simplex virus (HSV) and varicella-zoster virus (VZV) infections. Ganciclovir is used to treat
cytomegalovirus infections in immunocompromised patients. Ribavirin is used to treat
respiratory syncytial virus (RSV) infections in newborns. Combinations of nucleoside analogs
such as zidovudine (AZT), didanosine (ddl), and lamivudine (3TC) are used to treat HIV
infections.
Non-nucleoside polymerase inhibitors Primarily used to treat infections caused by herpesviruses. They inhibit the activity of viral
Foscarnet polymerases by binding to a site other than the nucleotide-binding site. Foscarnet is used
to treat ganciclovir-resistant cytomegalovirus (CMV) and acyclovir-resistant herpes simplex virus
(HSV).
Non-nucleoside reverse transcriptase inhibitors Used to treat HIV infections. They inhibit the activity of reverse transcriptase by binding to a
Nevirapine, delavirdine, efavirenz site other than the nucleotide-binding site and are often used in combination with nucleoside
analogs.
Genome Integration
Raltegravir Used to treat HIV infections.
Assembly and Release of Viral Particles
Protease inhibitors Used to treat HIV infections. They inhibit protease, an essential enzyme of HIV, by binding to
Indinavir, ritonavir, saquinavir, nelfinavir its active site.

Neuraminidase inhibitors Used to treat influenza virus infections.

Zanamivir, oseltamivir

Interfere with Viral Uncoating is incorporated into a growing nucleotide chain, it sometimes
prevents additional nucleotides from being added. In other cases,
After a virus enters a host cell, the nucleic acid must separate from
the analog results in a defective strand with altered base-pairing
the protein coat for replication to occur. Because of this, drugs that
properties. nucleotide, p. 32
interfere with the uncoating step prevent viral replication. Only
The basis for the selective toxicity of most nucleoside ana-
two drugs target this step—amantadine and rimantadine—and
logs is the fact that virally encoded enzymes are more likely than
they block influenza A viruses. They both prevent or reduce the
the host cell polymerases to incorporate a nucleotide analog.
severity of the disease, but viral strains develop resistance easily,
Therefore, more damage is done to the rapidly replicating viral
limiting their usefulness. influenza A virus, p. 508
genome than to the host cell genome. The analogs, however, are
only effective against replicating viruses. Viruses such as herpes-
virus and HIV can remain latent in cells, so the drugs do not cure
Interfere with Nucleic Acid Synthesis these infections; they simply shorten the active infection. Latent
Many of the most effective antiviral drugs take advantage of the virus can still reactivate, causing symptoms to recur.
error-prone, virally encoded enzymes that replicate viral nucleic Most nucleoside analogs are reserved for severe infections
acid. With few exceptions, however, these drugs are generally because of their significant side effects. An important exception is
limited to treating infections caused by herpesviruses or HIV. acyclovir, a drug used to treat herpesvirus infections. This drug
causes little harm to uninfected cells because normal cellular
Nucleoside Analogs enzymes do not convert the drug into a nucleotide analog. Instead,
A number of antiviral drugs are nucleoside analogs, compounds a virally encoded enzyme does this. The enzyme is only present in
similar in structure to a nucleoside. These analogs can be phos- cells infected by herpesviruses such as herpes simplex virus (HSV)
phorylated in vivo by a virally encoded or normal cellular enzyme and varicella-zoster virus (VZV). Other nucleoside analogs include
to form a nucleotide analog, a chemical structurally similar to ganciclovir, which is used to treat life- or sight-threatening cytome-
the nucleotides of DNA and RNA. When a nucleotide analog galovirus (CMV) infections in immunocompromised patients, and
476 Chapter 20 Antimicrobial Medications

ribavirin, which is used to treat respiratory syncytial virus infec- MicroAssessment 20.6
tions (RSV) in newborns. Nucleoside analogs used to treat HIV
infection interfere with the activity of reverse transcriptase and are Viral replication generally uses host cell machinery; because of this,
there are few targets for selectively toxic antiviral drugs. Available
called nucleoside reverse transcriptase inhibitors (NRTIs). antiviral drugs are virus-specific; targets include viral entry, viral
Unfortunately, the virus rapidly develops mutational resistance to uncoating, nucleic acid synthesis, integrase, and the assembly and
these drugs, which is why they are often used in combination with release of viral particles.
other anti-HIV drugs. NRTIs include zidovudine (AZT), didanos- 16. Explain why acyclovir has fewer side effects than do other
ine (ddI), and lamivudine (3TC). Two of these are often used in nucleoside analogs.
combination for HIV therapy. herpesviruses, pp. 535, 582 17. How do protease inhibitors interfere with the production of
infectious viral particles?
Non-Nucleoside Polymerase Inhibitors 18. Why are nucleoside analogs active only against replicating
Non-nucleoside polymerase inhibitors are compounds that inhibit viruses? +
the activity of viral polymerases by binding to a site other than
the nucleotide-binding site. One example, foscarnet, is used
to treat infections caused by ganciclovir-resistant CMV and 20.7 ■ Mechanisms of Action
acyclovir-resistant HSV.
of Antifungal Drugs
Non-Nucleoside Reverse Transcriptase
Learning Outcomes
Inhibitors (NNRTIs)
19. Describe the antifungal drugs that interfere with plasma
Non-nucleoside reverse transcriptase inhibitors (NNRTIs) inhibit membrane synthesis and function.
the activity of reverse transcriptase by binding to a site other than
20. Compare and contrast echinocandins, griseofulvin, and
the nucleotide-binding site. They are often used in combination flucytosine.
with nucleoside analogs to treat HIV infections. The medications
include nevirapine, delavirdine, and efavirenz. Eukaryotic pathogens such as fungi more closely resemble human
cells than do bacteria. This is why relatively few drugs are avail-
Prevent Genome Integration able for systemic use against fungal pathogens (table 20.3). The
targets of antifungal drugs are illustrated in figure 20.16.
Integrase inhibitors offer a new option for treating HIV infections.
They inhibit the HIV-encoded enzyme integrase, thereby prevent-
ing the virus from inserting the DNA copy of its genome into that Interfere with Plasma Membrane
of the host cell. Raltegravir is the first approved drug of this class. Synthesis and Function
HIV integrase, p. 698
The target of most antifungal drugs is ergosterol, a sterol found in
the plasma membrane of fungal but not human cells. sterol, p. 36
Prevent Assembly and Release
of Viral Particles Polyenes
Virally encoded enzymes required for the assembly and release of The polyenes, a group of antibiotics produced by certain spe-
viral particles are the targets of medications used to treat certain cies of Streptomyces, bind to ergosterol. This disrupts the fungal
viral infections.

Protease Inhibitors Cell division

Protease inhibitors are used to treat HIV infections. These medi- Griseofulvin
cations inhibit the HIV-encoded enzyme protease, which plays Plasma membrane
an essential role in the production of infectious viral particles. synthesis/function
When HIV replicates, several of its proteins are translated as a Polyenes
Azoles Nucleus
polyprotein, a single amino acid chain that must be cleaved by the Allylamines
protease to release the individual proteins. The various protease
inhibitors—including indinavir, ritonavir, saquinavir, and nelfina- Nucleic acid
synthesis
vir—differ in dosage and side effects. HIV protease, p. 698
Flucytosine

Neuraminidase Inhibitors Cell wall synthesis

Echinocandins
Neuraminidase inhibitors inhibit neuraminidase, an enzyme encoded
by influenza viruses. This enzyme is important for the release of
viral particles from infected cells. Two neuraminidase inhibitors are
currently available—zanamivir, which is administered by inhala- FIGURE 20.16 Targets of Antifungal Drugs
tion, and oseltamivir, which is taken orally. Both shorten the infec- ? What compound in the plasma membrane is the target of most
tion when taken within two days of the onset of symptoms. antifungal drugs?
 Part III Microorganisms and Humans 477

TABLE 20.3 Characteristics of Antifungal Drugs

Drug Target/Drug Comments

Plasma Membrane
Polyenes Bind to ergosterol, disrupting the plasma membrane and allowing the cytoplasm to leak out. Amphotericin B is
Amphotericin B, nystatin very toxic but the most effective drug for treating life-threatening infections; newer lipid-based emulsions are
less toxic but very expensive. Nystatin is too toxic for systemic use, but can be used topically.
Azoles Interfere with ergosterol synthesis, leading to defective cell membranes; active against a wide variety of fungi.
Imidazoles: ketoconazole, Used to treat a variety of systemic and localized fungal infections. Triazoles are less toxic than imidazoles.
miconazole, clotrimazole
Triazoles: fluconazole,
voriconazole

Allylamines Inhibit an enzyme in the pathway of ergosterol synthesis. Administered topically to treat dermatophyte
Naftifine, terbinafine infections. Terbinafine can be taken orally.

Cell Wall Synthesis

Echinocandins Interfere with β-1,3 glucan synthesis. Used to treat Candida infections as well as invasive aspergillosis that resists
Caspofungin other treatments.

Cell Division
Griseofulvin Used to treat skin and nail infections. Taken orally for months; concentrates in the dead keratinized layers of
the skin; taken up by fungi invading those cells and inhibits their division. Active only against fungi that invade
keratinized cells.
Nucleic Acid Synthesis
Flucytosine Used to treat systemic yeast infections; enzymes within yeast cells convert the drug to 5-fluorouracil, which
inhibits an enzyme required for nucleic acid synthesis; not effective against most molds; resistant mutants
are common.

membrane, killing the cell by allowing its cytoplasmic contents to Interfere with Cell Wall Synthesis
leak out. Unfortunately, the polyenes are quite toxic to humans,
Fungal cell walls contain some components not produced by
which limits their systemic use to life-threatening infections.
animal cells. Drugs in a family called echinocandins interfere
Amphotericin B causes severe side effects, but it is the most effec-
with synthesis of the fungal cell wall component β-1,3 glucan,
tive drug for treating certain systemic infections. Newer lipid-
causing fungal cells to burst. Caspofungin, the first member to be
based emulsions are less toxic but more expensive. Nystatin is too
approved, is used to treat Candida infections as well as invasive
toxic to be given systemically, but is used topically.
aspergillosis that resists other treatments.
Azoles
Interfere with Cell Division
The azoles are a family of chemically synthesized drugs, some
of which have antifungal activity. They include two classes—the The target of one antifungal drug, griseofulvin, is cell division.
imidazoles and the newer triazoles; the latter are generally less Griseofulvin interferes with the action of tubulin, a structure
toxic. Both classes inhibit ergosterol synthesis, resulting in defec- required for nuclear division. Because tubulin is a part of all
tive fungal membranes that leak cytoplasmic contents. Fluconazole eukaryotic cells, the selective toxicity of the drug may be due to
and voriconazole, which are triazoles, are increasingly being used its greater uptake by fungal cells. When taken orally for months,
to treat systemic fungal infections. Ketoconazole, an imidazole, it is absorbed and eventually concentrated in the dead keratinized
is also used systemically, but has more severe side effects. Other layers of the skin. Fungi that then invade keratin-containing struc-
imidazoles, including miconazole and clotrimazole, are com- tures such as skin and nails take up the drug, which prevents them
monly used in non-prescription creams, ointments, and supposi- from multiplying. Griseofulvin is only active against fungi that
tories to treat vaginal yeast infections. They are also applied to invade keratinized cells, and is used to treat skin and nail infec-
the skin to treat dermatophyte infections. dermatophyte, p. 543 tions. tubulin, p. 73 keratin, p. 336

Allylamines Interfere with Nucleic Acid Synthesis

The allylamines inhibit an enzyme in the pathway of ergosterol Nucleic acid synthesis is a common feature of all eukaryotic cells,
synthesis. Naftifine and terbinafine can be applied to the skin generally making it a poor target for antifungal drugs. The drug
to treat dermatophyte infections. Terbinafine can also be taken flucytosine, however, is taken up by yeast cells and then converted
orally. by yeast enzymes to an active, inhibitory form.
478 Chapter 20 Antimicrobial Medications

Flucytosine 20.8 ■ Mechanisms of Action

Flucytosine is a synthetic derivative of cytosine, one of the nucle-
obases. Enzymes within infecting yeast cells convert flucytosine
of Antiprotozoan and
to 5-fluorouracil, which inhibits an enzyme required for nucleic Antihelminthic Drugs
acid synthesis. Unfortunately, resistant mutants are common, and
therefore, flucytosine is used mostly in combination with ampho- Learning Outcome
tericin B or as an alternative drug for patients with systemic yeast 21. Describe the targets of most antiparasitic drugs.
infections who are unable to tolerate amphotericin B. Flucytosine
is not effective against molds. nucleobase, p. 32 Most antiparasitic drugs probably interfere with biosynthetic
pathways of protozoan parasites or the neuromuscular function
MicroAssessment 20.7 of worms. Unfortunately, compared with antibacterial, antifungal,
and antiviral drugs, little research and development goes into
Because fungi are eukaryotic cells, there are relatively few targets
for selectively toxic antifungal drugs. Most antifungal drugs interfere these drugs, because most parasitic diseases are concentrated in
with ergosterol function or synthesis. Other targets include cell wall the poorer areas of the world where people simply cannot afford
synthesis, cell division, and nucleic acid synthesis. to spend money on expensive medications.
19. Why is amphotericin B used only for treating life-threatening Some of the most important antiparasitic drugs and their char-
infections? acteristics are summarized in table 20.4.
20. Why is flucytosine generally used only in combination with
other drugs?

TABLE 20.4 Characteristics of Some Antiprotozoan and Antihelminthic Drugs

Causative Agent/Drug Comments
Intestinal protozoa
Iodoquinol Mechanism unknown; poorly absorbed but taken orally to eliminate amebic cysts in the intestine.
Nitazoxanide New drug used to treat cryptosporidiosis and giardiasis.
Nitroimidazoles Activated by the metabolism of anaerobic organisms. Interfere with electron transfer and alter DNA.
Do not reliably eliminate the cyst stage. Metronidazole is also used to treat infections caused by
Metronidazole
anaerobic bacteria.
Quinacrine Mechanism of action is unknown, but may be due to interference with nucleic acid synthesis.
Plasmodium (Malaria) and Toxoplasma
Folate antagonists Interfere with folate metabolism; used to treat toxoplasmosis and malaria.
Pyrimethamine, sulfonamide
Malarone A synergistic combination of atovaquone and proguanil hydrochloride used to treat malaria.
Atovaquone interferes with mitochondrial electron transport while proguanil disrupts folate
synthesis. The combination is active against both the blood stage and early liver stage of Plasmodium
species.
Quinolones The mechanism of action is not completely clear. Chloroquine is concentrated in infected red blood
Chloroquine, mefloquine, cells and is the drug of choice for preventing or treating the red blood cell stage of the malarial
primaquine, tafenoquine parasite. Its effects may be due to inhibition of an enzyme that protects the parasite from the toxic
by-products of hemoglobin degradation. Primaquine and tafenoquine destroy the liver stage of the
parasite and are used to treat relapsing forms of malaria. Mefloquine is used to treat infection caused
by chloroquine-resistant strains of the malarial parasite.
Trypanosomes and Leishmania
Eflornithine Used to treat infections caused by some types of Trypanosoma; inhibits the enzyme ornithine
decarboxylase.
Heavy metals These inactivate sulfhydryl groups of parasitic enzymes, but are very toxic to host cells as well.
Melarsoprol, sodium stibogluconate, Melarsoprol is used to treat trypanosomiasis, but the treatment can be lethal. Sodium stibogluconate
meglumine antimonate and meglumine antimonate are used to treat leishmaniasis.
Nitrofurtimox Widely used to treat acute Chagas’ disease; forms reactive oxygen radicals that are toxic to the
parasite as well as the host.
(continued)
 Part III Microorganisms and Humans 479

TABLE 20.4 Characteristics of Some Antiprotozoan and Antihelminthic Drugs (Continued )

Causative Agent/Drug Comments
Intestinal and Tissue Helminths
Avermectins Ivermectin causes neuromuscular paralysis in parasites; used to treat infections caused by
Ivermectin Strongyloides and tissue nematodes.
Benzimidazoles Mebendazole binds to tubulin of helminths, blocking microtubule assembly and inhibiting glucose
Mebendazole, thiabendazole, uptake. It is poorly absorbed in the intestine, making it effective for treating intestinal, but not tissue,
albendazole helminths. Thiabendazole may have a similar mechanism, but it is well absorbed and has many toxic
side effects. Albendazole is used to treat tissue infections caused by Echinococcus and Taenia solium.
Phenols Absorbed by cestodes in the intestinal tract, but not by the human host.
Niclosamide
Piperazines Piperazine causes a flaccid paralysis in worms and can be used to treat Ascaris infections.
Piperazine, diethylcarbamazine Diethylcarbamazine immobilizes filarial worms and alters their surface, which enhances killing by the
immune system. The resulting inflammatory response, however, causes tissue damage.
Pyrazinoisoquinolines A single dose of praziquantel is effective in eliminating a wide variety of trematodes and cestodes. It
Praziquantel is taken up but not metabolized by the worm, ultimately causing sustained contractions of the worm.
Tetrahydropyrimidines Pyrantel pamoate interferes with neuromuscular activity of worms, causing a type of paralysis. It is
Pyrantel pamoate, oxantel not readily absorbed from the gastrointestinal tract and is active against intestinal worms including
pinworm, hookworm, and Ascaris. Oxantel can be used to treat Trichuris infections.

FUTURE CHALLENGES 20.1

War with the Superbugs
With respect to antimicrobial drugs, the future keep at least one step ahead of bacterial resis- Various defensins and related compounds are
challenge is already upon us. The challenge is tance. Another method to foil drug resistance is being intensively studied as promising antimi-
to maintain the effectiveness of antimicrobials to interfere with the resistance mechanisms, as crobials.
by (1) preventing the continued spread of is done currently in using β-lactamase inhibi- Knowing the nucleotide sequences of
resistance, and (2) developing new drugs that tors in combination with β-lactam drugs to pathogens is allowing researchers to iden-
have even more desirable properties. New protect the antimicrobial drug from enzymatic tify genes associated with pathogenicity. This
ways must be developed to fight infections destruction. Alternatively, perhaps chemicals information, in turn, might help researchers
caused by the ever greater numbers of bacte- can be used to inactivate or interfere with bac- design antimicrobials that interfere directly
rial strains that are resistant to the effects terial efflux systems. with those processes. The “master switch”
of conventional antimicrobial drugs. Several Some researchers are focusing on devel- that controls expression of virulence determi-
strategies are being used to develop potential oping medications entirely unrelated to con- nants is one such potential target. Nucleotide
weapons against these “superbugs.” ventional antimicrobials. One example is a sequence information and determination of the
One way to combat resistance is to con- class of compounds called defensins, which three-dimensional conformation of proteins
tinue developing modifications of existing are short peptides, approximately 29 to 35 may uncover new targets for antimicrobial
antimicrobial drugs. Researchers constantly amino acids in length, produced naturally by a drug therapy.
work to modify drugs chemically, trying to variety of eukaryotic cells to fight infections.

Summary
20.1 ■ History and Development Development of New Generations of Drugs
of Antimicrobial Drugs Antimicrobial drugs can be chemically modified to give them new
Discovery of Antimicrobial Drugs properties (figure 20.1).
Salvarsan, developed by Paul Ehrlich, was the first documented exam- 20.2 ■ Features of Antimicrobial Drugs
ple of an antimicrobial medication. Most modern antibiotics come from species of Streptomyces and
Discovery of Antibiotics Bacillus (bacteria) and Penicillium and Cephalosporium (fungi).
Alexander Fleming discovered that a species of the fungus
Penicillium produces a drug he called penicillin.
480 Chapter 20 Antimicrobial Medications

Selective Toxicity Antibacterial Medications That Interfere

Medically useful antimicrobials are selectively toxic; the relative toxic- with Metabolic Pathways (figure 20.8)
ity of a drug is expressed as the therapeutic index. Sulfa drugs competitively inhibit an enzyme in the metabolic pathway
that leads to folic acid synthesis. Trimethoprim inhibits the enzyme that
Antimicrobial Action catalyzes a metabolic step following the one inhibited by sulfonamides.
Bacteriostatic drugs inhibit the growth of bacteria; drugs that kill bac-
Antibacterial Medications That Interfere
teria are bactericidal.
with Cell Membrane Integrity
Spectrum of Activity Daptomycin damages the cytoplasmic membrane. Polymyxin B dam-
Broad-spectrum antimicrobials affect a wide range of bacteria; those ages membranes of Gram-negative bacteria.
that affect a narrow range are called narrow-spectrum. Antibacterial Medications Effective Against
Effects of Combinations Mycobacterium Species
Combinations of antimicrobial drugs can be synergistic, antagonistic, First-line drugs that specifically target Mycobacterium species include
or additive. isoniazid, ethambutol, and pyrazinamide. Second-line drugs are used
for strains resistant to the first-line drugs.
Tissue Distribution, Metabolism,
and Excretion of the Drug 20.4 ■ Determining the Susceptibility of a Bacterial
Some antimicrobials cross the blood-brain barrier into the cerebrospinal Strain to an Antimicrobial Drug
fluid; these can be used to treat meningitis. Drugs that are unstable in Minimum Inhibitory and Bactericidal Concentrations
acid must be administered through injection. Drugs that have a long (MIC and MBC) (figure 20.9)
half-life need to be administered less frequently. The minimum inhibitory concentration (MIC) is the lowest concen-
Adverse Effects tration of a specific antimicrobial drug needed to prevent the growth of
a bacterial strain in vitro. The minimum bactericidal concentration
Some people develop allergies to certain antimicrobials. Some antimi-
(MBC) is the lowest concentration of a specific antimicrobial drug that
crobials can have potentially damaging side effects such as kidney dam-
kills 99.9% of cells of a given strain of bacterial in vitro.
age. When the composition of the normal microbiota is altered, which
happens when a person takes antimicrobials, pathogens normally unable Conventional Disc Diffusion Method (figure 20.10)
to compete may grow to high numbers. The Kirby-Bauer disc diffusion test determines the susceptibility of a
bacterial strain to a battery of antimicrobial drugs.
Resistance to Antimicrobials
Certain types of bacteria have intrinsic (innate) resistance to a par- Commercial Modifications of Antimicrobial
ticular drug. Acquired resistance is due to spontaneous mutation or the Susceptibility Testing
acquisition of new genetic information. Automated methods can determine antimicrobial susceptibility in as
little as 4 hours (figure 20.11).

20.3 ■ Mechanisms of Action of Antibacterial Drugs 20.5 ■ Resistance to Antimicrobial Drugs (figure 20.13)
(figure 20.2, table 20.1)
Mechanisms of Acquired Resistance (figure 20.14)
Antibacterial Medications That Inhibit Cell Enzymes that chemically modify a drug render it ineffective. Structural
Wall Synthesis (figure 20.3) changes in the target can prevent the drug from binding. Altered porin
The β-lactam drugs, which include the penicillins, cephalosporins, proteins prevent drugs from entering cells. Efflux pumps actively
carbapenems, and monobactams, irreversibly inhibit penicillin- pump drugs out of cells.
binding proteins (PBPs), ultimately leading to cell lysis (figure  20.4). Acquisition of Resistance
Vancomycin binds to the peptide side chain of NAM, blocking
Resistance can be acquired through spontaneous mutation or horizontal
peptidoglycan synthesis. Bacitracin interferes with the transport of
gene transfer. The most common mechanism of transfer of antibiotic
peptidoglycan precursors across the cytoplasmic membrane; it is a
resistance genes is through the conjugative transfer of R plasmids.
common ingredient in non-prescription ointments.
Examples of Emerging Antimicrobial Resistance
Antibacterial Medications That Inhibit
Vancomycin-resistant enterococci (VRE), methicillin-resistant
Protein Synthesis (figure 20.7)
Staphylococcus aureus (MRSA), vancomycin-intermediate S. aureus
Antibiotics that inhibit protein synthesis by binding to the 70S ribo- (VISA), vancomycin-resistant S. aureus (VRSA), penicillin-resistant
some include the aminoglycosides, the tetracyclines, the macrolides, Streptococcus pneumoniae, multidrug-resistant Mycobacterium
chloramphenicol, the lincosamides, the oxazolidinones, and the tuberculosis (MDR-TB), extensively drug-resistant Mycobacterium
streptogramins. tuberculosis (XDR-TB), and carbapenem-resistant Enterobacteriaceae
(CRE) are all examples of emerging antimicrobial resistance.
Antibacterial Medications That Inhibit Nucleic
Acid Synthesis Slowing the Emergence and Spread of Antimicrobial
The fluoroquinolones interfere with DNA replication and transcription Resistance
by inhibiting one or more topoisomerases. The rifamycins block pro- Physicians should prescribe antimicrobial medications only when
karyotic RNA polymerase from initiating transcription. Metronidazole appropriate. The public must be educated about the appropriateness and
is activated by anaerobic metabolism, and the activated form then binds limitations of antimicrobial therapy. Patients need to carefully follow
DNA. prescribed instructions when taking antimicrobials.
 Part III Microorganisms and Humans 481

20.6 ■ Mechanisms of Action of Antiviral Drugs 20.7 ■ Mechanisms of Action of Antifungal Drugs 
(figure 20.15, table 20.2) (figure 20.16, table 20.3)

Prevent Viral Entry Interfere with Plasma Membrane Synthesis and Function
Enfuvirtide and maraviroc prevents HIV from entering cells. The polyenes disrupt fungal cell membranes by binding to ergosterol.
The azoles inhibit the synthesis of ergosterol. The allylamines inhibit an
Interfere with Viral Uncoating enzyme in the pathway of ergosterol synthesis.
Amantadine and rimantadine block the uncoating of influenza A virus
after it enters a cell. Interfere with Cell Wall Synthesis
Echinocandins interfere with the synthesis of β-1,3 glucan.
Interfere with Nucleic Acid Synthesis
Interfere with Cell Division
Most antiviral drugs take advantage of the error-prone virally encoded
enzymes used to replicate viral nucleic acid. Nucleoside analogs are Griseofulvin is concentrated in keratinized skin cells, where it inhibits
phosphorylated in vivo to form nucleotide analogs; when these are fungal cell division.
incorporated into viral DNA they interfere with replication. Interfere with Nucleic Acid Synthesis
Prevent Genome Integration Flucytosine is taken up by yeast cells and converted by yeast enzymes
Raltegravir inhibits HIV integrase. to an active form.

Prevent Assembly and Release of Viral Particles 20.8 ■

Mechanisms of Action of Antiprotozoan
Protease inhibitors bind to and inhibit protease, the enzyme required for and Antihelminthic Drugs (table 20.4)
the production of infectious HIV particles. Neuraminidase inhibitors Most antiparasitic drugs are thought to interfere with biosynthetic path-
interfere with the release of influenza virus particles from a cell. ways of protozoan parasites or the neuromuscular function of worms.

Review Questions
Short Answer 2. Penicillin has been modified to make derivatives that differ in all
1. Describe the difference between the terms antibiotic and of the following except
antimicrobial. a) spectrum of activity.
2. Define therapeutic index and explain its importance. b) resistance to β-lactamases.
3. Explain the role of penicillin-binding proteins in drug suscepti- c) potential for allergic reactions.
bility. d) a and c.
4. Name three classes of antimicrobial drugs that target ribosomes. 3. Which of the following is the target of β-lactam antibiotics?
5. Explain the roles of the first-line drugs versus the second-line a) Peptidoglycan synthesis b) DNA synthesis
drugs in the treatment of tuberculosis. c) RNA synthesis d) Protein synthesis
6. Compare and contrast the method for determining the minimum e) Folic acid synthesis
inhibitory concentration (MIC) with the Kirby-Bauer disc diffu- 4. Which of the following statements is false?
sion test.
a) A bacteriostatic drug stops the growth of a microorganism.
7. Name three targets that can be altered sufficiently via spontaneous
b) The lower the therapeutic index, the less toxic the drug.
mutation to result in resistance to an antimicrobial drug.
c) Broad-spectrum antibiotics are associated with the development
8. What is MRSA? Why is it significant? of Clostridium difficile–associated disease.
9. Why is it difficult to develop antiviral drugs? d) Azithromycin has a longer half-life than does penicillin V.
10. Explain the difference between the mechanism of action of an e) Chloramphenicol can cause a life-threatening type of anemia.
azole and that of a polyene. 5. All of the following interfere with the function of the ribosome
except
Multiple Choice a) fluoroquinolones. b) lincosamides. c) macrolides.
1. Which of the following targets would you expect to be the most d) streptogramins. e) tetracyclines.
selective with respect to toxicity? 6. The target of the sulfonamides is
a) Cytoplasmic membrane function a) cytoplasmic membrane proteins.
b) DNA synthesis b) folate synthesis.
c) Glycolysis c) gyrase.
d) Peptidoglycan synthesis d) peptidoglycan biosynthesis.
e) 70S ribosome e) RNA polymerase.
482 Chapter 20 Antimicrobial Medications

7. Routine antimicrobial therapy to treat tuberculosis involves taking Applications

a) one drug for 10 days. 1. A physician was treating one young woman and one elderly patient
b) two or more drugs for 10 days. for urinary tract infections caused by the same type of bacterium.
c) one drug for at least 6 months. Although the patients had similar body dimensions and weight, the
d) two or more drugs for at least 6 months. physician gave a smaller dose of drug to the older patient. What
e) five drugs for 2 years. was the physician’s rationale for this decision?
8. Staphylococcus aureus strains referred to as HA-MRSA are sensi- 2. An advocacy group in Washington, D.C., is petitioning the U.S.
tive to Department of Agriculture (USDA) to stop the use of low-dosage
a) methicillin. b) penicillin. c) cephalosporin. antimicrobial agents used to enhance the growth of cattle and
chickens. Why is the group against this practice? Why does the
d) vancomycin. e) none of the above.
USDA permit it?
9. Acyclovir is a
a) nucleoside analog. b) non-nucleoside polymerase inhibitor. Critical Thinking +
c) protease inhibitor. d) none of the above. 1. Figure 20.12 shows the E-test procedure for determining an MIC
10. The antifungal drug griseofulvin is used to treat value. How would the zone of inhibition appear if the drug concen-
a) vaginal infections. b) systemic infections. trations in the strip were decreased slightly?
c) nail infections. d) eye infections. 2. Why is acyclovir converted to a nucleotide analog only in cells
infected with herpes simplex virus?
21 Respiratory System Infections
KKE EYYT TE ERRMMS S
Antigenic Drift Minor changes
that occur naturally in influenza virus
antigens as a result of mutation.
Antigenic Shift Major changes
Mucociliary Escalator Layer of
mucus moved by cilia lining the
respiratory tract that traps bacteria
and other particles and moves them
into the throat.
in the antigenic composition of
influenza viruses that result from Multidrug-Resistant
reassortment of viral nucleic acids Tuberculosis (MDR-TB) Strains of
during infection of the same host cell Mycobacterium tuberculosis resistant
by different viral strains. to isoniazid and rifampicin—two of
the first-line anti-TB drugs.
Croup Acute obstruction of the
larynx occurring mainly in infants Otitis Media Infection of the
and young children, often resulting middle ear.
from respiratory syncytial or other Pharyngitis Inflammation of the
viral infection. throat.
Directly Observed Therapy Pneumonia Inflammation of the
Short-Course (DOTS) Method lungs accompanied by filling of the
used to ensure that patients comply air sacs with fluids such as pus and
with their tuberculosis treatment; blood.
the healthcare worker watches
while the patient takes each dose of Sputum Pus and other material
medication. coughed up from the lungs.
Extensively Drug-Resistant Tubercle Granuloma formed
Individual with streptococcal pharyngitis. Tuberculosis (XDR-TB) Strains of in tuberculosis; granulomas are
Mycobacterium tuberculosis resistant collections of lymphocytes and
to the first-line anti-TB drugs macrophages found in a chronic
isoniazid and rifampicin, and at least inflammatory response, an attempt
A Glimpse of History three of the second-line TB drugs. by the body to wall off and contain
persistent organisms and antigens.
Rebecca Lancefield’s mother was a direct descendent of Lady Mary
Wortley Montagu, who promoted smallpox variolation in England more
than 75 years before Edward Jenner. Lancefield attended Wellesley
College in 1912, and became interested in biology. She was awarded a
scholarship to Columbia University, where she studied under the famous
microbiologist Hans Zinsser, a pioneer in the science of immunology. from human infections had the same cell wall carbohydrate—“A”—
She received a master’s degree, but her studies were interrupted when whereas streptococci from other sources had different wall carbohydrates:
her husband was drafted into the armed services in World War I. “B” from cattle infections; “C” from cattle, horses, and guinea pigs; “D”
Fortunately he was assigned to the Rockefeller Institute, where Rebecca from cheese and human normal microbiota; and so on. The “Lancefield
got a job as a laboratory technician for the distinguished microbiologists grouping” proved to be a much better predictor of pathogenic poten-
O. T. Avery and A. R. Dochez. These scientists were known for their tial than hemolysis on blood agar. Lancefield became the first woman
research on pneumococci, and were studying other streptococci from president of the American Association of Immunologists and in 1970
personnel at army camps. At that time, classification of streptococci was elected to the prestigious National Academy of Sciences. She died in
was largely based on whether their colonies produced colorless clearing 1981 at the age of 86.
(β-hemolysis) or a greenish partial clearing (α-hemolysis) when grown
on blood agar. Avery and Dochez showed that streptococci could be

R
espiratory infections include an enormous variety of ill-
more effectively divided into groups based on their surface antigens.
nesses ranging from the trivial to the fatal. They can be
Mary Wortley Montagu, p. 419
divided into infections of the upper respiratory system
The Lancefields returned to Columbia University, where Rebecca
received her Ph.D. in 1925 for her studies of α-hemolytic streptococci.
(in the head and neck) and infections of the lower respiratory
Thereafter, she went back to the Rockefeller Institute where she spent system (in the chest). Upper respiratory infections such as colds
the rest of her scientific career studying streptococci. She showed that are extremely common and uncomfortable, but they are not
β-hemolytic streptococci could be classified according to cell wall car- life-threatening and go away without treatment in about a week.
bohydrates, a system now known as “Lancefield grouping.” Lancefield Diseases of the lower respiratory system such as pneumonia and
demonstrated that almost all strains of β-hemolytic streptococci isolated tuberculosis are often serious, however, and may be fatal.

483
484 Chapter 21 Respiratory System Infections

21.1 ■ Anatomy, Physiology, projections—along their exposed free border. The cilia beat
synchronously, continually propelling the mucus film out of the
and Ecology respiratory tract. The mucus, along with any entrapped microbes,
is then swallowed and digested. This mechanism, the mucociliary
Learning Outcomes escalator, normally keeps the lower respiratory tract completely
1. Outline the functions of the upper and lower respiratory tracts. free of microorganisms. Smoking, alcohol and narcotic abuse,
2. List the parts of the respiratory system that are normally and viral infections all impair ciliary movement and increase the
microbe-free. chance of infection. cilia, p. 74
The tonsils are secondary lymphoid organs, located so that
A critical function of the respiratory system is gas exchange. Each they come into contact with microbes entering the upper respira-
breath brings in O2, which replenishes the supply in the blood, tory tract. They are important in the immune response but can also
and then releases CO2, the waste product of cellular metabolism. be the sites of infection, resulting in tonsillitis. secondary lymphoid
In addition, movement of the vocal cords makes sound, which organs, p. 358
allows us to speak, and sensors in the nose detect odors. Although Some of the bacterial genera that inhabit the upper respiratory
the mouth can also be used for breathing, it is considered part of system are listed in table 21.1. Although generally harmless, they
the digestive tract, so it is covered in chapter 24. are opportunists that can cause disease when host defenses are
The respiratory system has two general parts—the upper impaired. opportunists, p. 382
respiratory tract and the lower respiratory tract. We also include
the eyes and ears in our discussion. Even though these are not The Nose and Nasal Cavity
considered part of the respiratory tract, the eyes and the nose are Air enters the respiratory system at the nostrils and flows into
important from a microbiological standpoint because they serve the nasal cavity. When cold air enters the nose, the blood flow to
as two main portals of entry to the body. Pathogens often first the tissues there immediately increases due to nervous reflexes,
establish themselves there and then spread to other parts of the warming the air to near body temperature and saturating it with
body. portals of entry, p. 439 water vapor.
The nasal entrance usually contains diphtheroids and staph-
ylococci. In addition, about 20% of healthy people constantly
The Upper Respiratory Tract carry Staphylococcus aureus in their noses; an even higher
The upper respiratory tract includes the nose and nasal cavity, percentage of hospital personnel are likely to be carriers of this
pharynx (throat), and epiglottis (figure  21.1). Mucous mem- important hospital- or community-acquired pathogen. Further
branes line the respiratory tract. These are coated with mucus, inside the nasal passages, the normal microbiota are similar to
a slimy glycoprotein material that traps air-borne dust and other that of the throat. Infection of the nasal passages, usually by
particles, including microorganisms. Mucus is produced by spe- viruses, results in rhinitis; this inflammation of the nasal tissues
cialized cells called goblet cells that are scattered among the cells causes the familiar “runny nose.”
of the membrane. Their name reflects their shape, which is narrow
at the base and wide at the surface, like a wine glass or goblet. Pharynx (Throat) and Epiglottis
mucous membrane, p. 337 Once past the nose, the air moves down to the throat (pharynx).
Ciliated epithelium lines most of the mucous membranes Inflammation of the throat, pharyngitis, is commonly the result of
of the respiratory system. These cells have cilia—tiny, hairlike viral infection. In addition to being part of the respiratory system,

TABLE 21.1 Normal Microbiota of the Upper Respiratory System

Genus Characteristics Comments

Staphylococcus Gram-positive cocci in clusters Facultative anaerobes. Commonly includes the potential pathogen
Staphylococcus aureus, inhabiting the nostrils.
Corynebacterium Pleomorphic, Gram-positive rods; Aerobic or facultatively anaerobic. Diphtheroids include anaerobic and
non-motile; non-spore-forming aerotolerant organisms.
Moraxella Gram-negative diplococci and diplobacilli Aerobic. Some microscopically resemble pathogenic Neisseria species
such as N. meningitidis.
Haemophilus Small, Gram-negative rods Facultative anaerobes. Commonly include the potential pathogen
H. influenzae.
Bacteroides Small, pleomorphic, Gram-negative rods Obligate anaerobes.
Streptococcus Gram-positive cocci in chains Aerotolerant (obligate fermenters). α (especially viridans, meaning
green hemolysis), β (clear hemolysis), and γ (non-hemolytic) types; the
potential pathogen, S. pneumoniae is often present.
 Part IV Infectious Diseases 485

FIGURE 21.1 Anatomy and Infections of the

Respiratory System (a) Lateral view of upper
respiratory tract. (b) Frontal view of the upper and Columnar
lower respiratory tracts. epithelium
? Why would inflammation in the lungs be Secretory
(goblet) cell
life-threatening?
Mucus
Cilia
Frontal sinus Flow of mucus

Adenoviral pharyngitis
Common cold
Diphtheria Sphenoid sinus
Ear infections
Epiglottitis
Adenoids
Laryngitis
Strep throat
Eustachian
Tonsillitis
tube opening
Opening of
Nasolacrimal maxillary sinus
duct opening

Tonsil
Pharynx
Epiglottis
Larynx
(a)

Frontal sinus
Nasal chamber
Upper
Nasolacrimal duct
Respiratory
Middle ear
System
Mastoid air cell
Bronchioles
Eustachian tube Alveoli
Maxillary sinus (air sacs)
Pharynx

Trachea Capillaries

Bronchi

Pleura
Lower
Respiratory
System

Arterioles

Lungs
(b) Bronchiolitis Legionnaires’ disease
Bronchitis Pleurisy
Coccidioidomycosis Pneumonia
Hantavirus pulmonary syndrome RSV infections
Histoplasmosis Tuberculosis
Influenza Whooping cough
486 Chapter 21 Respiratory System Infections

the throat is essential to the digestive system. A small muscular flap, the eustachian tubes. The inner ear, which is fluid-filled, is also
the epiglottis, covers the opening to the lower respiratory tract dur- microbe-free. Occasionally, however, viruses or bacteria enter
ing swallowing, preventing material from entering it. Inflammation the inner ear, often following an upper respiratory tract infec-
of the epiglottis, called epiglottitis, can be a life-threatening emer- tion, causing labyrinthitis (named for the chambers of the inner
gency because the swollen flap can block the airway. ear which are called labyrinths). The skull also has air-filled
Streptococci, including viridans streptococci (a group chambers—the sinuses and mastoid air cells. Infections of these
of α-hemolytic streptococci) and non-hemolytic species, are chambers are called sinusitis and mastoiditis, respectively.
common members of the normal microbiota of the throat. A vari-
ety of other bacteria are also found there, including Moraxella The Lower Respiratory Tract
catarrhalis, diphtheroids, and anaerobic Gram-negative bacteria,
such as Bacteroides species. In addition, a number of opportunistic The lower respiratory tract includes the larynx (voice box),
pathogens frequently colonize the throat, including Streptococcus trachea, bronchi, and lungs (figure  21.1b). Inflammation of the
pneumoniae, Haemophilus influenzae, and Neisseria meningitidis. larynx is called laryngitis, and is manifest as hoarseness. The
α-hemolysis, p. 96 trachea (windpipe) is a continuation of the larynx and branches
into two bronchi. Inflammation of the bronchi is called bronchitis,
MicroByte commonly the result of viral infection or smoking. The bronchi
Cilia beat at a rate of about 1,000 times a minute, propelling mucus branch repeatedly, becoming bronchioles, the site of an important
along the mucociliary escalator. viral infection called bronchiolitis. The smallest branches of the
bronchioles end in the alveoli, which are tiny, thin-walled air
Eyes sacs that make up the bulk of lung tissue. Inflammation of the
lungs is called pneumonitis, often the result of viral infections.
The surface of the eyes and lining of the eyelids are covered by
Pneumonitis that causes the alveoli to fill with pus and fluid is
mucous membranes called conjunctiva. Infection of the conjunc-
called pneumonia. Lung tissues have many macrophages that
tiva is called conjunctivitis. The tear ducts connect the eyes to the
readily move into the alveoli and airways to engulf infectious
nasal chamber. Infection of the tear ducts is called dacryocystitis.
agents, helping to prevent pneumonia from developing. The lungs
Surprisingly, even though the eyes are constantly exposed
are surrounded by two membranes called pleura: One adheres to
to large numbers of microorganisms, the conjunctiva of healthy
the lung and the other to the chest wall and diaphragm. The pleura
people commonly have very few bacteria. Presumably, this is
normally slide against each other as the lung expands and con-
because the eye is bathed with lysozyme-rich tears and cleaned
tracts. Inflammation of the pleura is called pleurisy, characterized
by the eyelid’s blinking reflex, which wipes the eye surface like a
by severe chest pain. The lower respiratory tract is usually sterile
car windshield wiper cleans a windshield. Unless microbes on the
and has no normal microbiota.
conjunctiva are able to attach to it, they are swept into the tear duct
and nasopharynx. The few bacteria found on the normal conjunc-
MicroAssessment 21.1
tiva usually originate from the skin microbiota and are generally
unable to colonize the respiratory system. lysozyme, p. 62 The respiratory system provides a warm, moist environment for
microorganisms. It is protected by tonsils and adenoids, and by the
Ears mucociliary escalator. The upper respiratory tract contains highly
diverse microbiota, including aerobes, anaerobes, facultative anaerobes,
The ear has three parts: external, middle, and inner ear. The exter- and aerotolerant bacteria. Although most of them are of low virulence,
nal ear is protected from microbes by cerumen (ear wax). The these organisms can sometimes cause disease opportunistically. The
middle ear is sterile. It is connected by the eustachian tubes to the lower respiratory system is free of a normal microbiota.
nasopharynx. These tubes equalize the pressure in the middle ear 1. What is the normal function of the tonsils and adenoids?
and drain normal mucus secretions. They are also a route through 2. Describe the normal microbiota of the respiratory system.
which microbes can enter the middle ear, leading to infection
3. How would paralysis of the cilia impact the respiratory
called otitis media. Enlargement of the adenoids can contribute system? +
to middle ear infections by interfering with normal drainage from

INFECTIONS OF THE UPPER RESPIRATORY SYSTEM

21.2 ■ Bacterial Infections of the
Upper Respiratory System A number of bacterial species can infect the upper respiratory
tract. Some, such as Haemophilus influenzae, can cause sore
Learning Outcomes throats but generally do not require treatment because the bacteria
3. Compare the distinctive characteristics of strep throat and diphtheria. are quickly eliminated by the immune system. Others, such as
Streptococcus pyogenes, cause infections that require treatment
4. List the parts of the upper respiratory system commonly infected by
Streptococcus pneumoniae and Haemophilus influenzae. because they are not as easily eliminated and can cause serious
complications.
 Part IV Infectious Diseases 487

Streptococcal Pharyngitis
(“Strep Throat”)
Sore throat is one of the most common reasons that
people in the United States seek medical care. Many
of these visits are due to justifiable concerns about
streptococcal pharyngitis, commonly known as strep
throat. One concern about streptococcal infections is
the risk of post-streptococcal sequelae. These are com-
plications that develop after the initial infection and will
be discussed in the next section.

Signs and Symptoms FIGURE 21.3 Streptococcus pyogenes Colonies on Blood

Strep throat is characterized by a sore throat, difficulty swallow- Agar The colonies are surrounded by a wide zone of β-hemolysis.
ing, and fever. The throat is red, with patches of pus and scattered ? What causes β-hemolysis?
tiny hemorrhages. The lymph nodes in the neck are enlarged and
tender. Abdominal pain or headache may occur in older children
and young adults. Patients do not usually have a cough, weepy
eyes, or runny nose. Most patients with strep throat recover spon- species. S. pyogenes is characterized by the “A” carbohydrate in its
taneously after about a week. In fact, many infected people have cell wall. Antibodies that bind to the group A carbohydrate are the
only mild symptoms or no symptoms at all. basis for many of the rapid diagnostic tests done on throat specimens
in a physician’s office (see figure 18.9).
Causative Agent Different S. pyogenes strains within GAS are distinguished
Strep throat is caused by Streptococcus pyogenes, a Gram-positive by variations of a surface antigen called M protein, an important
coccus that grows in chains (figure  21.2). The organism can be virulence factor. Strains with certain types of M protein are the
differentiated from other streptococci that normally inhabit the cause of strep throat, whereas others with different M protein are
throat by its colony morphology on blood agar—S. pyogenes colo- more likely to cause skin infections.
nies are surrounded by a characteristic clear zone of β-hemolysis
(figure  21.3). In contrast, most species of Streptococcus that Pathogenesis
are typically part of the normal throat microbiota are either Streptococcus pyogenes has many virulence factors (table 21.2).
α-hemolytic, producing a zone of greenish partial clearing around Some of these disease-causing mechanisms are structural com-
colonies on blood agar, or non-hemolytic. A few other strepto- ponents of the cell wall that allow the bacterium to avoid host
cocci and some other bacteria are also β-hemolytic, so further
tests are needed to identify S. pyogenes. hemolysis, p. 96 blood
agar, p. 95
Streptococcus pyogenes is commonly referred to as the group A Virulence Factors of
TABLE 21.2
streptococcus (GAS), reflecting its Lancefield grouping (see Streptococcus pyogenes
A Glimpse of History in this chapter). This grouping system uses Product Effect
antibodies to distinguish the cell wall carbohydrates in streptococcal
C5a peptidase Inhibits recruitment of phagocytes by
destroying complement C5a
Hyaluronic Inhibits phagocytosis
FIGURE 21.2 acid capsule
Streptococcus pyogenes
M protein Interferes with phagocytosis by causing
Chain formation of
breakdown of complement C3b, an opsonin
S. pyogenes, revealed
by fluorescence Protein F Responsible for attachment to host cells
microscopy.
Protein G Binds to Fc portion of antibody, thereby
? What are two interfering with opsonization
sequelae that may
Streptococcal Superantigens responsible for scarlet fever,
occur after strep
pyrogenic toxic shock, “flesh-eating” fasciitis
throat?
exotoxins
(SPEs)
Streptolysins O Lyse leukocytes and erythrocytes
and S
Tissue- Enhance spread of bacteria by breaking
degrading down DNA, proteins, blood clots, tissue,
enzymes hyaluronic acid
10 μm
488 Chapter 21 Respiratory System Infections

phagocytosis) from being deposited on the bacterial cell wall.

Another cell wall protein—protein G—is an Fc receptor that binds
M protein the Fc portion of IgG, preventing opsonization by antibodies (see
figure 16.11). The organism releases C5a peptidase, an enzyme
Protein G that destroys complement component C5a, normally responsible
Protein F
for attracting phagocytes to the site of a bacterial infection, thus
further inhibiting phagocytosis. It also produces streptolysins O
Lipoteichoic acid and S—enzymes that destroy erythrocytes and leukocytes by mak-
ing holes in their cell membranes. Leukocyte destruction inhibits
the immune response, whereas destruction of erythrocytes causes
the β-hemolysis exhibited by S. pyogenes. Some S. pyogenes strains
have a hyaluronic acid capsule that further prevents phagocytosis.
Hyaluronic
Hyaluronic acid is a normal component of human tissue, so the cap-
acid capsule sule is thought to be a cloaking disguise, making it difficult for the
host immune system to detect the pathogen. complement system, p. 344
C5a peptidase, p. 389 membrane-damaging toxins, p. 392 opsonins,
p. 347

Peptidoglycan A few strains of S. pyogenes produce streptococcal pyrogenic

exotoxins (SPEs), a family of exotoxins that cause severe strep-
tococcal diseases characterized by high fever (pyro means “fire”).
SPEs are encoded by bacteriophages, an example of lysogenic
Group A conversion. These toxins are superantigens, causing massive acti-
carbohydrate vation of T cells. The resulting uncontrolled release of cytokines
is probably responsible for the seriousness of these infections. A
Cytoplasmic
membrane person with strep throat caused by an SPE-producing strain of
S. pyogenes may develop scarlet fever, characterized by high
fever, roughening of the skin (texture like sandpaper) and a pink-
red rash. Although the toxin that causes these symptoms is an SPE,
it is traditionally referred to as erythrogenic toxin (erythro means
“red”). The rash of scarlet fever is found on the head, neck, chest
and thighs, but usually not around the mouth. The toxin also
causes the tongue to look like a ripe strawberry—red and spot-
Streptococci
ted. Both the skin and the tongue may peel. Some SPE-producing
FIGURE 21.4 Virulence Factors on the Cell Envelope of strains of S. pyogenes have additional virulence factors that allow
Streptococcus pyogenes them to cause severe invasive diseases such as streptococcal
? What is the role of M protein in pathogenesis? toxic shock syndrome, and “flesh-eating” necrotizing fasciitis.
lysogenic conversion, p. 313 superantigens, p. 393 cytokines, p. 341
necrotizing fasciitis, p. 553 streptococcal toxic shock syndrome, p. 553
defenses (figure 21.4). Others are destructive enzymes and toxins
released by the bacterial cell that damage or kill host cells. MicroByte
Proteins in the cell wall of S. pyogenes allow the bacteria to Streptococcal streptokinase is used as a clot-dissolving medication
attach to host cells. M protein is an important adhesin involved in treating some heart attacks and pulmonary embolisms.
in attachment—antibodies that bind to it prevent infection. There
are more than 80 antigenic types of M protein among the many
strains of S. pyogenes, however, and antibodies to one type do Epidemiology
not prevent infection by a strain that has a different kind. Another Streptococcus pyogenes naturally infects only humans. The strains
protein, protein F, mediates attachment of S. pyogenes to cells that cause strep throat spread easily by respiratory droplets gener-
of the throat by adhering to fibrin, a protein found on epithelial ated by shouting, coughing, and sneezing. Epidemics have also
cells. originated from food contaminated with S. pyogenes. Nasal car-
Once S. pyogenes colonizes host tissues, it produces enzymes riers of S. pyogenes are more likely than pharyngeal carriers to
such as DNase, hyaluronidase, and proteases that break down spread the organisms. Anal carriers are not common but can be
intracellular connections and allow the organism to spread rap- a dangerous source of healthcare-associated infections. A person
idly to other cells. This spread is assisted by streptokinase, which may be an asymptomatic carrier of S. pyogenes for weeks, if they
causes the breakdown of blood clots. are not given treatment. Some people become long-term carriers;
S. pyogenes has many mechanisms for avoiding the host in these cases, the infecting strain usually becomes deficient in
immune system. M protein, in addition to aiding attachment to the M protein and is not a threat to the carrier or to others. The peak
host cell, also interferes with phagocytosis. It prevents comple- incidence of strep throat occurs in winter or spring and is highest
ment component C3b (an opsonin that would usually increase in grade school children. healthcare-associated infections, p. 449
 Part IV Infectious Diseases 489

Treatment and Prevention pains, chest pains, rash, and nodules under the skin. Uncontrollable
People with fever and sore throat should be taken to a physician body movements (chorea, commonly called St. Vitus’ dance) can
so that a throat swab can be taken for a rapid diagnostic test and occur and, when present, are major criteria for diagnosis. Carditis,
throat culture. Confirmed strep throat is treated with a full 10 days the most serious complication, develops in about a third to a half
of penicillin or erythromycin, which eliminates the organism in of patients. This can lead to chronic rheumatic heart disease in
about 90% of the cases. Treatment given as late as 9 days after the which one or more of the heart valves are damaged, causing them
onset prevents post-streptococcal sequelae (covered next). to leak and resulting in heart failure later in life. The damaged
Adequate ventilation and avoiding crowded situations help valves are also prone to infection, usually by bacteria from the
to control the spread of streptococcal infections. No vaccine is normal skin or mouth microbiota, resulting in subacute bacterial
available. However, genome sequencing has revealed some new endocarditis. subacute bacterial endocarditis, p. 673

possibilities for vaccine targets. Table  21.3 summarizes some Rheumatic fever only develops in people who are geneti-
important facts about strep throat. cally predisposed to the disease—some MHC class II alleles are
involved in susceptibility (see Perspective 15.1). The pathogenesis
of rheumatic fever is not well understood but is thought to be an
Post-Streptococcal Sequelae autoimmune response involving both humoral and cell-mediated
Post-streptococcal sequelae—complications that can develop after immunity—in some individuals, antibodies to S. pyogenes cross
strep throat or other streptococcal infections—are thought to be a react with host tissue antigens that are similar to the pathogen
result of immune responses to Streptococcus pyogenes. They are antigens, a case of molecular mimicry. In chronic rheumatic
uncommon in Western industrialized nations because of quick heart disease, for example, molecular mimicry between cardiac
identification and treatment of S. pyogenes infections. Globally, myosin and epitopes of streptococcal M protein may result in
however, they occur much more frequently and can be serious. production of antibodies that bind to both S. pyogenes M protein
and host cardiac myosin. If such antibodies bind to cardiac tissue,
Acute Rheumatic Fever the tissue is then targeted for attack by the host’s own effector
Acute rheumatic fever usually begins about 3 weeks after recov- T helper (TH ) cells. Several other shared antigens have also been
ery from strep throat. Signs and symptoms include fever, joint identified. This recognition of self-antigens starts an autoimmune

TABLE 21.3 Strep Throat (Streptococcal Pharyngitis)

1 Streptococcus pyogenes enters Signs and Sore, red throat, with pus and tiny
by inhalation (nose), or by symptoms hemorrhages, enlargement and
ingestion (mouth). 6
tenderness of lymph nodes in the neck;
2 Pharyngitis, fever, enlarged 1 less frequently, abscess formation
lymph nodes; sometimes involving tonsils; occasionally, rheumatic
3 2 2 fever and glomerulonephritis as sequelae
tonsillitis, abscess; scarlet fever
with strains that produce Incubation 2 to 5 days
erythrogenic toxin. 5 period
Symptoms go away. 7
Causative agent Streptococcus pyogenes, Lancefield
S. pyogenes exits by nose group A β-hemolytic streptococci
3
and mouth. 4 5 Pathogenesis Virulence associated with hyaluronic acid
Late complications appear: capsule and M protein, both of which
inhibit phagocytosis; protein G binds
4 Glomerulonephritis Fc segment of IgG; protein F for mucosal
5 Rheumatic fever attachment; multiple enzymes.

6 Neurological abnormalities Epidemiology Direct contact and droplet infection;

ingestion of contaminated food.
Complications subside.
Damaged heart valves leak, Treatment and Treatment: 10 days of penicillin or
7
heart failure develops. prevention erythromycin.
5
Prevention: Avoiding crowds; adequate
ventilation; daily penicillin to prevent
recurrent infection in those with a history
of rheumatic heart disease.
490 Chapter 21 Respiratory System Infections

response—the T cells release proinflammatory cytokines, causing Puffiness around the

eyes (fluid retention) Streptococcus
inflammation that leads to permanent damage to the local tissues, pyogenes
particularly the heart valves. MHC, p. 369 inflammation, p.  348 antigen Immune
complex
APC, p. 369 helper T cells, p. 356 molecular mimicry, p. 412 Antibody
Although outbreaks of rheumatic fever still occur in the Complement
United States, incidence has generally declined. This is probably
a result of quick treatment of strep throat with antibiotics, and Glomerulus
decreased prevalence of the strains associated with the disease.
Overall, the risk of developing acute rheumatic fever after severe
untreated strep throat is 3% or less, and those with untreated mild Kidney
pharyngitis have an even lower risk. Nonetheless, 10 to 20 million Glomerulus
new cases of rheumatic fever occur globally each year, mostly in
economically disadvantaged countries. Signs and symptoms of
rheumatic fever usually subside with rest and anti-inflammatory
medicines such as aspirin. Individuals with cardiac damage due Bladder
to acute rheumatic fever take penicillin daily for years to prevent
Nephron
recurrence of the disease.

Acute Post-Streptococcal Glomerulonephritis

Acute post-streptococcal glomerulonephritis occasionally fol- FIGURE 21.5 Pathogenesis
lows strep throat but is more often an aftermath of a strep skin of Glomerulonephritis Immune
infection. It begins 7–21 days after the initial infection, usually complexes are deposited in the
about 10 days in the case of strep throat. The signs and symptoms kidney glomeruli, causing an
Swollen ankles inflammatory response.
include fever, fluid retention, high blood pressure, and blood and (fluid retention)
protein in the urine (making it look like tea or coca cola). There ? What is the role of nephrons
in the kidney?
are no streptococci in the urine or the diseased kidney tissues—
S. pyogenes has generally been eliminated from the throat by the
body’s immune response by the time the symptoms of glomerulo- and throat or in the nasal cavity. Heart and kidney failure and
nephritis appear. The damage to the kidneys is due to an inflam- paralysis may occur later.
matory reaction caused by streptococcal antigens that accumulate
in the kidney glomeruli (tufts of tiny blood vessels and nephrons, Causative Agent
responsible for urine formation); antibodies are bound to these Diphtheria is caused by Corynebacterium diphtheriae, a pleomor-
antigens and these immune complexes activate the comple- phic, non-motile, non-spore-forming, Gram-positive rod that often
ment system (figure 21.5). Only a few of the many strains of stains irregularly. The organisms are club-shaped (koryne means
S. pyogenes cause the condition, and it is rare to get glomerulone- “club”) and often occur side by side in “palisades” (like a wooden
phritis twice. immune complexes, p. 395 fence) (figure 21.6).

Diphtheria
Diphtheria, a deadly toxin-mediated disease, is now rare in
the United States because of childhood immunization. Events
in other parts of the world, however, are a reminder of what
can happen when public health is neglected. In 1990, a diph-
theria epidemic began in the Russian Federation. Over the next
5 years, it spread to all the newly independent states of the
former Soviet Union. By the end of 1995, 125,000 cases and
4,000 deaths had been reported. The social and economic dis-
ruption following the breakup of the Soviet Union had allowed
diphtheria to reemerge after being well controlled over the
previous quarter of a century.

Signs and Symptoms

Diphtheria usually begins with a mild sore throat and slight fever, FIGURE 21.6 Corynebacterium diphtheriae
with extreme fatigue and malaise. Dramatic swelling of the neck ? How do strains of C. diphtheriae acquire the gene for toxin
occurs. A whitish-gray pseudomembrane forms on the tonsils production?
 Part IV Infectious Diseases 491

Most strains of C. diphtheriae release diphtheria toxin, a process, so one or two molecules of it can inactivate nearly all the
powerful exotoxin responsible for the serious symptoms of the cell’s EF-2. This explains the extreme potency of diphtheria toxin.
disease. The gene for this toxin is carried by a specific lysogenic A-B toxins, p. 392 endocytosis, p. 72
bacteriophage, another example of lysogenic conversion.
To isolate C. diphtheriae from throat material, a special
Epidemiology
medium that contains potassium-tellurite is used. This chemical
inhibits most of the normal microbiota of the upper respiratory Humans are the primary reservoir for Corynebacterium diphtheriae.
tract and causes C. diphtheriae to form black colonies, aiding in The organisms are typically spread by air from infected people.
identification. The organism can also be grown on Loeffler’s, a They are then acquired either by inhalation or from fomites.
medium that enhances volutin granule formation at the poles of C. diphtheriae can sometimes cause cutaneous diphtheria and the
the cells, which can be seen with methylene blue stains. lysogenic chronic skin ulcers that develop may become a source of infection
conversion, p. 313 volutin granules, p. 67
to others. This problem primarily occurs in homeless populations.
fomites, p. 440

Pathogenesis
Corynebacterium diphtheriae has little invasive ability, and Treatment and Prevention
rarely enters the blood or tissues. The disease is caused by the Diphtheria is treated by injecting the patient with antiserum
diphtheria exotoxin released by the bacteria growing in the against diphtheria toxin as soon as possible. Delaying treatment
throat. The pseudomembrane that forms is made up of dead while waiting for culture results can be fatal, so the antiserum
epithelial cells (killed by the exotoxin) and clotted blood, must be given immediately once the disease is suspected. The
along with fibrin and the leukocytes that accumulate during bacteria are sensitive to antibiotics such as erythromycin and peni-
inflammation. This membrane may come loose and obstruct cillin, but such treatment only stops transmission of the disease; it
the airways, causing the patient to suffocate. The toxin may has no effect on toxin that has already been absorbed. Even with
be absorbed into the bloodstream, allowing it to access and treatment, about 1 of 10 diphtheria patients dies. antiserum, p. 420
damage heart, nerve, and kidney tissues. Because diphtheria results from toxin production rather than
Diphtheria toxin is an A-B toxin (figure 21.7). The B subunit microbial invasion, it can be effectively prevented by immuni-
attaches to specific receptors on a host cell membrane, and the zation with toxoid. The toxoid, prepared by formalin treatment
entire toxin molecule is taken into the cell by endocytosis. Cells of diphtheria toxin, causes the body to produce antibodies that
lacking these receptors do not take up the toxin and are unaffected specifically neutralize the toxin. The childhood vaccination DTaP
by it—this receptor specificity explains why some tissues of the consists of diphtheria and tetanus toxoids along with components
body are not affected in diphtheria, while others are severely dam- of Bordetella pertussis. Unfortunately, these immunizations
aged. Once the toxin molecule is inside the cell, the A subunit are often neglected, and serious epidemics of the diseases have
separates from the B subunit. It becomes an active enzyme and occurred periodically. Since the 1980s, there has been an active
catalyses a chemical reaction that inactivates elongation factor 2 campaign in most of the United States to ensure that children who
(EF-2), which is required for movement of the eukaryotic ribo- are entering school are immunized against diphtheria. As a result,
some on mRNA. This stops protein synthesis, and the cell dies. the incidence of the disease has been reduced to only a few cases
Since the toxin A subunit is an enzyme, it is not used up in the a year. Immunity decreases after childhood, however, so booster

A subunit
A 3 Toxin subunits 4 Enzyme A enters cytoplasm
B separate and A
B subunit and inactivates a protein in the
Receptor enters the 80S ribosomes, stopping
cytoplasm. protein synthesis and causing
cell death.
1 A-B toxin attaches
by B subunit to 2 Toxin enters cell
membrane receptor by endocytosis.
of susceptible cell.

Endosome Ribosome

FIGURE 21.7 Mode of Action of Diphtheria Toxin

? Why is diphtheria toxin so potent?
492 Chapter 21 Respiratory System Infections

TABLE 21.4 Diphtheria

1 Corynebacterium diphtheriae Signs and Sore throat, fever, fatigue, and malaise;
enters by inhalation. symptoms pseudomembrane forms on tonsils and
2 Infection established in nasal throat or in nose; paralysis, heart and
cavity and/or throat. 1 3 kidney failure

3 Toxin released, 5 2
6
pseudomembrane forms. 4 Incubation 2 to 6 days
period
4 Toxin causes paralysis, 4
damages heart muscle, Causative agent Corynebacterium diphtheriae, an A-B
kidneys, nerves. toxin–producing, non-spore-forming
Gram-positive rod
5 Membrane may come loose
4 Pathogenesis Infection in upper respiratory tract;
and obstruct breathing.
exotoxin is released and absorbed
6 Exit from body by respiratory by bloodstream; toxin kills cells by
secretions. interfering with protein synthesis; effect
is on cells that have receptors for the
toxin—mainly heart, kidney, and nerve
tissue.
Epidemiology Inhalation of infectious droplets; direct
contact with patient or carrier; indirect
contact with contaminated articles.
Treatment and Treatment: antitoxin; erythromycin to
prevention prevent transmission.
Prevention by immunization with
diphtheria toxoid; given to children at
6 weeks, 4 months, 6 months, 18 months,
and 4 to 6 years in DTaP vaccine;
boosters every 10 years.

injections must be given every 10 years to maintain protection. severe malaise also occur. A thick green nasal discharge that may
Table  21.4 summarizes some important facts about diphtheria. contain pus and blood sometimes develops as well.
toxoid, p. 423
Causative Agents
Pinkeye, earache, and sinus infections are often caused by two
Pinkeye, Earache, and Sinus Infections common bacterial pathogens: (1) Haemophilus influenzae, a tiny
Bacterial infections of the eye surface (conjunctivitis or “pink-
eye”), middle ear (otitis media), and sinuses (sinusitis) are very
common, often occur together, and often have the same caus-
ative agent. Otitis media is especially common in children, and
is responsible for 30 million doctor visits per year in the United
States, at an estimated cost of $1 billion. Pinkeye is easily spread
and is therefore a concern when it occurs in a daycare facility or
school. Sinusitis is common in both adults and children.

Signs and Symptoms

The signs and symptoms of acute bacterial conjunctivitis include
increased tears, redness of the conjunctiva, swollen eyelids, sensi-
tivity to bright light, and large amounts of pus (figure 21.8). Acute
bacterial conjunctivitis differs from viral conjunctivitis, in which
eyelid swelling and pus are usually minimal.
Otitis media manifests with severe earache—in a typical
case, a young child wakes from sleep screaming with pain. The
intense pain often causes vomiting. Fever is generally mild or FIGURE 21.8 Bacterial Conjunctivitis Redness, swollen eyelids,
absent. and pus, characteristic of bacterial conjunctivitis.
In sinusitis, facial pain and a pressure sensation characteris- ? What symptom shown here is usually minimal in viral
tically occur in the region of the involved sinus. Headache and conjunctivitis?
 Part IV Infectious Diseases 493

Gram-negative rod; and (2) Streptococcus pneumoniae, the Gram- Infected middle ear
positive encapsulated diplococcus known as the pneumococcus. Eardrum
Strains that infect the conjunctiva have adhesins that allow them (bulging)
to attach firmly to the epithelium. Haemophilus influenzae, p.  646
Streptococcus pneumoniae, p. 497 adhesins, p. 386
Conjunctivitis can also be caused by a variety of other organ-
isms, including Moraxella lacunata, enterobacteria and Neisseria
gonorrhoeae. Although not very common, some eye infections are
caused by environmental microbes that contaminate eye medica- External
tions and contact lens solutions. When these infections occur they ear canal
are usually serious and can lead to permanent eye damage. People
who wear contact lenses should follow the instructions
on use of cleansing solutions exactly; cloudy or outdated
solutions and eye medications should be thrown away. Pus
Otitis media and sinusitis can also be caused by
Mycoplasma pneumoniae, Streptococcus pyogenes,
Eustachian
Moraxella catarrhalis, and Staphylococcus aureus. About tube (inflamed)
one-third of the cases are caused by respiratory viruses,
explaining why some infections do not respond to anti- Ventilation tube
biotics, which have no effect on viruses.

Pathogenesis
Few details are known about the pathogenesis of acute
bacterial conjunctivitis. The organisms are probably inoc- FIGURE 21.9 Otitis Media Inset shows a ventilation tube placed
in the eardrum to equalize middle ear pressure in individuals with
ulated directly onto the conjunctiva from airborne respiratory chronically malfunctioning eustachian tubes.
droplets or rubbed in from contaminated hands. Like most bacte-
rial pathogens, they resist destruction by lysozyme. Attachment is ? Why is otitis media painful?

aided in some cases by degradation of mucin, a protective compo-

nent of epithelial surface mucus. Following attachment, the bacte- A preceding or simultaneous viral illness is common in otitis
ria release proteases, collagenases, and coagulases, combined with media and sinusitis; the virus probably damages the mucociliary
toxins in some cases, that further damage the tissue and allow the mechanism that would normally protect against bacterial infection.
entry of the organisms. lysozyme, p. 62 Otitis media is rare in the first month of life, but it becomes very
Otitis media and sinusitis are usually preceded by infection common in early childhood. Children who use pacifiers beyond
of the nasal chamber and nasopharynx that probably spreads the age of 2 years have an increased risk of developing otitis
upward through the eustachian tube (figure  21.9). The infec- media. Conditions that cause inflammation of the nasal mucosa—
tion damages the ciliated cells, resulting in inflammation and including viral infections, nasal allergies, exposure to air pollution and
swelling. Because the damaged eustachian tube cannot move cigarette smoke—play a role in some cases. Older children develop
secretions from the middle ear, fluid and pus collect behind immunity to H. influenzae, and the bacterium rarely causes otitis
the eardrum. This leads to a buildup of pressure, causing the media in children beyond age five. Sinusitis tends to affect adults and
ear to ache. The eardrum may perforate (burst), discharging older children in whom the sinuses are more fully developed.
blood or pus from the ear and giving immediate relief from
pain. With treatment, holes in the eardrum usually heal quickly. Treatment and Prevention
The pressure in the middle ear can force infected material into Bacterial conjunctivitis is effectively treated with eyedrops or
the mastoid air cells, resulting in mastoiditis. The fluid behind ointments containing an antibacterial medication to which the
the eardrum may also impair hearing ability, resulting in some infecting strain is sensitive. Antibacterial therapy with amoxicillin
young children showing a delay in speech development. Both is generally effective against both otitis media and sinusitis; alter-
middle ear and sinus infections sometimes spread to the brain native medications are available for communities where antibiotic-
coverings, causing meningitis. meningitis, p. 643
resistant strains of H. influenzae and S. pneumoniae are common.
In general, antibiotics have been used indiscriminately in treating
Epidemiology otitis media. However, when properly used, they decrease the
The ecological factors involved in the appearance and spread of risk of serious complications such as mastoiditis and meningitis.
the eye, ear, and sinus infections caused by H. influenzae and Decongestants and antihistamines generally are ineffective and can
S. pneumoniae are largely unknown; carrier rates can sometimes be harmful because they reduce the immune response.
reach 80% in the absence of disease. The virulence of the bacte- General preventive measures for conjunctivitis include hand-
ria, crowding, and presence of respiratory viruses are probably all washing, and protecting the eyes from contamination by avoid-
important factors in these epidemics. ing rubbing or touching them, particularly with shared towels.
494 Chapter 21 Respiratory System Infections

Bacterial conjunctivitis is highly contagious, so individuals sus- Signs and Symptoms

pected of having it are kept home from school or daycare settings Colds begin with malaise, followed by a scratchy or mildly sore
for diagnosis and start of treatment. Otitis media during the “flu” throat, runny nose, cough, and hoarseness. The nasal secretions
season can be substantially decreased by giving influenza vaccine are initially profuse and watery, then thicken in a day or two,
to infants in daycare facilities. Ampicillin or sulfasoxazole given finally becoming cloudy and greenish. There is no fever unless
continuously over the winter and spring are useful preventives secondary bacterial infection occurs. Symptoms are mostly gone
in people who have three or more cases of otitis media within a within a week, but a mild cough sometimes continues for longer.
6-month period. Surgical removal of enlarged adenoids improves
drainage from the eustachian tubes and can help prevent recur- Causative Agents
rences in certain patients. In those with chronically malfunc-
Viruses that cause the common cold are often referred to simply
tioning eustachian tubes and hearing loss, tiny plastic tubes are
as “cold viruses.” Between 30% and 50% of colds are caused by
often inserted through the eardrums so that pressure can equalize
the 100 or more types of human rhinoviruses (rhino means “nose,”
(figure  21.9). There are no proven preventive measures for
as in rhinoceros, or “horny nose”) (figure 21.10). These are mem-
sinusitis.
bers of the picornavirus family (pico means “small” and rna is
ribonucleic acid; thus, “small RNA viruses”), a group of naked
MicroAssessment 21.2
viruses that have a single-stranded RNA genome. Rhinoviruses
Streptococcus pyogenes, a group A streptococcus with many can usually be grown in cell cultures under temperature and pH
virulence factors, causes a sore throat commonly known as strep
conditions that mimic the upper respiratory tract (33°C and at a
throat. Untreated S. pyogenes infections can sometimes cause serious
diseases (sequelae) in other parts of the body long after the initial slightly acid pH). They are inactivated if the pH drops below 5.3,
infections have resolved. In diphtheria, toxin is absorbed into the and therefore are usually destroyed in the stomach. Many other
bloodstream and circulates throughout the body, selectively damaging viruses and some bacterial species can also produce the signs and
certain tissues such as heart, kidneys, and nerves. Conjunctivitis, otitis symptoms of the common cold.
media, and sinusitis are common infections that often occur together
and are caused by the same pathogens. Pathogenesis
4. Name two post-streptococcal sequelae. Rhinoviruses attach to specific receptors on respiratory epithelial
5. How does diphtheria toxin kill cells? cells and then infect these cells. The replication cycle produces
6. How would adequate ventilation help to prevent the spread of large numbers of virions and these are released to infect other
streptococcal infections? + cells. Ciliary motion in the infected cells stops and the cells
may die and slough off. The damage causes the release of pro-
inflammatory cytokines and stimulates nervous reflexes, result-
ing in increased nasal secretions, tissue swelling that partially
21.3 ■ Viral Infections of the or completely obstructs the airways, and sneezing. Later in the
Upper Respiratory System inflammatory response, blood vessels dilate, allowing plasma to
ooze out and leukocytes to migrate to the infected area. Secretions
Learning Outcomes from the area may then contain pus and blood. The infection is
5. List the strategies helpful in avoiding common colds. eventually stopped by the innate and adaptive responses, but it can
6. Give the distinctive characteristics of adenoviral pharyngitis. spread into the ears, sinuses, or even the lower respiratory tract

The average person in the United States gets two to five viral
upper respiratory infections each year. Although hundreds of dif-
ferent viruses cause these infections, the range of symptoms they
produce is similar. These infections generally resolve without any
treatment and rarely cause permanent damage. However, they Rhinoviruses
impair respiratory tract defenses and allow for more serious sec-
ondary bacterial infections.
Host cell
The Common Cold
The common cold is the most frequent infectious disease in
humans, and accounts for more than half of the upper respiratory
tract infections that people get every year. Colds are the lead-
ing cause of absences from school, and result in people missing
150 million workdays per year.

MicroByte
The average adult gets between 2 and 4 common colds a year, FIGURE 21.10 Rhinovirus-Infected Cell (TEM)
whereas children can get as many as 8 colds in this time.
? What kinds of cells do rhinoviruses infect?
 Part IV Infectious Diseases 495

before this occurs. Rhinoviruses can even cause life-threatening

TABLE 21.5 The Common Cold
pneumonia in individuals with AIDS.
Signs and Scratchy throat, nasal discharge, malaise,
symptoms headache, cough
Epidemiology
Incubation 1 to 2 days
Humans are the only source of cold viruses, which are spread
period
by close contact with an infected person. In adults the disease
is usually contracted when airborne virus-containing droplets Causative agent Mainly rhinoviruses—more than 100 types;
many other viruses, some bacteria
are inhaled. Transmission can also occur when secretions from
infected people are accidently rubbed into the eyes or nose by Pathogenesis Viruses attach to respiratory epithelium,
starting infection that spreads to adjacent
contaminated hands. Viruses introduced into the eye quickly enter
cells; ciliary action ceases and cells
the nasal passage via the nasolacrimal duct. A person with severe slough; mucus secretion increases, and
symptoms early in the course of a cold is much more likely to inflammatory reaction occurs; infection
transmit the virus than is someone with mild symptoms or in the stopped by interferon release, cell-
late stage of the disease. This is because infected people have mediated and humoral immunity.
very high concentrations of virus in their nasal secretions and on Epidemiology Inhalation of infected droplets; transfer
their hands during the first 2 or 3 days of a cold. By the fourth or of infectious mucus to nose or eye by
fifth day, virus levels are often undetectable, but low levels can contaminated fingers; children initiate many
outbreaks in families because of lack of care
be present for 2 weeks. Although a few virions are sufficient to with nasal secretions.
infect the nasal mucosa, colds are actually not highly contagious
Treatment and No generally accepted treatment except
if reasonable preventive measures such as handwashing are taken.
prevention for control of symptoms. Handwashing;
In a study in which non-immune adults were exposed to infected avoiding people with colds and touching
individuals, less than half contracted colds. Young children, how- face.
ever, transmit cold and other respiratory viruses very effectively
because they are often careless with their respiratory secretions.
Experimental and epidemiological studies show that there is no
relationship between exposure to low temperatures and develop- Signs and Symptoms
ment of colds, contrary to popular belief. Emotional stress, how-
Adenoviruses that infect the upper respiratory tract generally
ever, can almost double the risk of catching a cold.
cause a runny nose, but unlike the common cold, fever is typically
present. The throat is usually sore, with regions of gray-white
Treatment and Prevention pus on the pharynx and tonsils, signs and symptoms that may be
There are no proven treatments for the common cold. Like all confused with those of strep throat. The lymph nodes of the neck
viruses, rhinoviruses are not affected by antibiotics or other anti- become large and tender and a mild cough is common. In some
bacterial medications. Analgesics (painkillers) and antipyretics epidemics, conjunctivitis is a prominent symptom. Diarrhea may
(fever-reducers) such as aspirin and ibuprofen can help reduce occur. Patients sometimes develop a severe cough with chest pain
symptoms. However, experimental evidence suggests that these (symptoms that can be confused with pneumonia), whooping
drugs somewhat prolong symptoms and duration of virus excre- cough, or pleurisy. Recovery usually takes about 1 to 3 weeks.
tion, and delay antibody production and thus recovery. pleurisy, p. 486
Mechanisms to prevent the spread of rhinoviruses include
handwashing (even in plain water) to physically remove the Causative Agent
viruses, keeping hands away from the face, and avoiding crowds
More than 50 antigenic types of adenoviruses infect humans.
and crowded places like subways when respiratory diseases are
The viruses are non-enveloped, with double-stranded DNA. Like
prevalent. It is especially important to avoid people with colds
many other naked viruses, they can remain infectious in the envi-
during the first few days of their symptoms, when they are shed-
ronment for long periods of time and are resistant to destruction
ding high numbers of viral particles. It has not been possible to
by detergents and alcohol solutions. They are easily inactivated,
develop a vaccine because such a large number of immunologi-
however, by heat (56°C), adequate levels of chlorine, and various
cally different viruses cause colds. However, all known rhinovirus
other disinfectants.
strain genomes have been sequenced, revealing new possibilities
for vaccines. Table 21.5 summarizes some facts about the com-
mon cold. Pathogenesis
Adenoviruses infect epithelial cells by attaching to receptors
near the basement membrane. Once inside the cell, the genome
Adenoviral Respiratory Tract Infections is transported to the host cell nucleus, where the virus multiplies.
Adenoviruses are widespread and can cause a variety of different The virus has many mechanisms for avoiding host defenses,
types of infections, depending on the viral serotype. For example, including delaying apoptosis, blocking interferon function, and
some cause a sore throat, whereas others cause eye infection. They interfering with antigen presentation by MHC class I molecules.
are representative of the many viruses that cause febrile (meaning Once replication is complete, a virally encoded “death protein”
with fever) upper respiratory tract infections. is produced that causes host cell lysis. In severe infections,
496 Chapter 21 Respiratory System Infections

chlorinated swimming pools. Adenoviral disease is spread by

TABLE 21.6 Adenoviral Pharyngitis
respiratory droplets so it can be a problem in military recruits
Signs and Fever, very sore throat, severe cough, and groups living together in crowded conditions. Asymptomatic
symptoms swollen lymph nodes of neck, pus on tonsils infections are common, which fosters epidemic spread. The
and throat, sometimes conjunctivitis; less
viruses are shed from the respiratory tract during the acute illness
frequently, pneumonia
and continue to be eliminated in the feces for months thereafter.
Incubation 5 to 10 days
period
Treatment and Prevention
Causative agent Adenoviruses—more than 45 types
As with colds, there is no specific treatment for adenoviral dis-
Pathogenesis Virus multiplies in host cells; cell ease, and most patients recover on their own. Secondary bacterial
destruction and inflammation occur; infections may occur, however, and these require treatment with
different types produce different
symptoms.
an antibacterial medication.
An attenuated vaccine administered orally was helpful for pre-
Epidemiology Inhalation of infected droplets; possible
venting acute respiratory disease in military recruits, but the vaccine
spread from gastrointestinal tract.
program was abandoned in 1996 because of cost. Now, with more
Treatment and No treatment except for relief of than 2,500 adenoviral illnesses monthly and some deaths, reinstitu-
prevention symptoms. No vaccine. Avoided by
handwashing, avoiding people with
tion of adenoviral vaccination of recruits is planned. Vaccines for
symptoms. two strains are currently under evaluation. Table 21.6 summarizes
some important facts about adenoviral pharyngitis.

MicroAssessment 21.3
extensive cell destruction and inflammation occur. Different types
of adenoviruses affect different tissues. basement membrane, p. 336 Many different kinds of infectious agents cause upper respiratory tract
apoptosis, p. 350 interferon, p. 341 MHC class I molecule, p. 369 diseases, often with the same signs and symptoms. Emotional stress
significantly increases the risk of contracting the common cold, but
exposure to cold temperatures probably does not. People with a cold
Epidemiology are most likely to transmit it if symptoms are severe, and during the
Humans are the only reservoir of adenoviruses. Because these first few days of illness. Adenovirus infections resemble colds, but
viruses can survive in the environment, healthcare personnel fever is present.
must take precautions to prevent transferring them from one 7. Why are there no vaccines for the common cold?
patient to another on medical instruments. Adenoviruses com- 8. How is an adenovirus infection treated?
monly infect schoolchildren, usually resulting in sporadic cases, 9. People who work at polar ice stations often do not develop colds.
but occasionally causing outbreaks in winter and spring. Summer Is this an expected observation? Why or why not? +
epidemics can occur when viruses are transmitted in inadequately

INFECTIONS OF THE LOWER RESPIRATORY SYSTEM

21.4 ■ Bacterial Infections of the healthcare-associated infections. Whooping cough, tuberculosis,
and Legionnaires’ disease are other serious infections of the lungs.
Lower Respiratory System
Learning Outcomes Pneumococcal Pneumonia
7. Compare the distinctive features of pneumococcal, Klebsiella, and Pneumococci are an important cause of community-acquired
mycoplasmal pneumonia. pneumonia, accounting for about 60% of the adult pneumonia
8. Outline the pathogenesis of pertussis and tuberculosis. patients requiring hospitalization (see Perspective 21.1).

Bacterial infections of the lower respiratory system are less com- Signs and Symptoms
mon than those of the upper respiratory system, mostly because The typical signs and symptoms of pneumococcal pneumonia are
they are stopped by the immune defenses at the portal of entry. cough, fever, chest pain, and sputum production (sputum is pus
However, they are generally much more serious. An earache or sore and other material coughed up from the lungs). These are usually
throat is unlikely to be life-threatening, but the causative organism preceded by a day or two of runny nose and upper respiratory
can cause serious illnesses if it infects the lungs. Pneumonias are congestion, ending with a sudden rise in temperature and a single,
inflammatory diseases of the lung in which fluid fills the alveoli. intense chill. The sputum quickly becomes pinkish or rust colored
They top the list of fatal community-acquired infections (meaning because it contains blood from the lungs. The severe chest pain
in the general population) in the United States, and are common is aggravated by each breath or cough, causing shallow, rapid
 Part IV Infectious Diseases 497

PERSPECTIVE 21.1
Terror by Mail: Inhalation Anthrax
During October and November 2001, 22 The spores are easy to produce and remain an attack, presumably because the best defense
human cases of anthrax were reported in the viable for years. When anthrax is acquired was considered an opponent’s fear of counter-
United States, half due to inhalation of Bacillus by inhalation, the fatality rate is very high, attack. As a result of the 2001 anthrax-by-mail
anthracis spores (inhalation anthrax) and half yet the disease is easily confined to the attack incident, a number of questions came into
due to skin infections (cutaneous anthrax). area because it does not spread person-to- focus: How do you diagnose anthrax quickly,
Five deaths occurred, all among the inhalation person. Bacillus anthracis spores were used as and what is the best way to teach and organ-
cases. More than 30,000 individuals potentially a weapon as early as World War I, although ize medical practitioners to meet an anthrax
exposed to the spores were given antibiotic ineffectively, in an attack on livestock used attack? What tests are available to identify
treatment as a preventive measure. Most of for food. Just before World War II, Japan, the B. anthracis rapidly and reliably, and what
the anthrax cases could reasonably be linked United States, USSR, Germany, and Great is the best way to sample people and the
to four mailing envelopes containing purified Britain secretly began to develop and perfect environment? How can decontamination of
B. anthracis spores, which contaminated peo- anthrax weapons, but they were not used buildings and other objects be accomplished?
ple, air, and surfaces during their journey during the war. During the “cold war” that Is there sufficient vaccine available, can bet-
through the postal system to their destinations. followed, both the United States and the ter vaccines be developed, and what is their
Sometimes called “anthrax pneumonia,” USSR developed massive biological warfare role before or after exposure to B. anthracis?
inhalation anthrax victims generally fail to programs that employed many thousands of What is the best antimicrobial treatment, is
show the typical symptoms of pneumonia. people, perfecting techniques for preparing the there enough of it, is it readily accessible,
Inhaled B. anthracis spores are quickly taken spores and delivering them to enemy targets. and how long should it be given to exposed
up by lung phagocytes and carried to the An executive order by President Nixon ended individuals? Can other potential treatments
regional lymph nodes, where they germi- the U.S. program in 1969, and the stockpiled such as antitoxins and designer medications to
nate, kill the phagocytes, and invade the weapons were ordered destroyed, but other block the lethal effect of B. anthracis toxin be
bloodstream. Although the organisms are sus- countries continued weapon development. In developed?
ceptible to various antibacterial medications, 1979, an accidental release of a tiny amount The mail attack has stimulated remark-
treatment must be given as soon as possible of B. anthracis spores from a biological war- able progress toward answering these ques-
after infection, because the bacteria produce fare plant in the USSR caused more than 90 tions, especially in the area of using the latest
a powerful toxin that usually kills the victim deaths downwind of the facility. The 1990 war technology for early detection, and teaching
within a day or two after bloodstream invasion with Iraq revealed their large anthrax weapon medical personnel about symptoms, signs, and
occurs. program, and several times during the 1990s a X-ray findings. Antibacterial treatment proves
Bacillus anthracis endospores have long Japanese terrorist group tried ineffectively to highly effective if given soon after exposure.
been considered for use in biological warfare. attack Tokyo institutions with anthrax spores. Approaches to combatting the toxin are under
The organism is readily available—anthrax During this long history of anthrax development, using designer drugs and anti-
is endemic in livestock in most areas of the weapon development, remarkably little was bodies. More information about anthrax is
world including the United States and Canada. accomplished that would help defend against available at www.mhhe.com/nester7.

breathing; the patient becomes short of breath

and develops a dusky color because of poor
oxygenation. People who survive without treat-
ment show profuse sweating and a rapid fall in
temperature to normal after 7 to 10 days.

Causative Agent
Pneumococcal pneumonia is caused by S. pneumoniae
Streptococcus pneumoniae, a Gram-positive
diplococcus known as pneumococcus (fig-
ure 21.11). The most striking characteristic of S.
pneumoniae is its thick polysaccharide capsule,
which is responsible for the organism’s virulence. Neutrophils
There are 90 different serotypes of S. pneu-
moniae, each with different capsular antigens.
Certain serotypes are more commonly associated
with pneumonia and other invasive diseases
(invasive means they infect normally sterile body FIGURE 21.11 Streptococcus pneumoniae Gram stain of
sites). Strains of the organism that lack a capsule sputum from a person with pneumococcal pneumonia.
do not cause invasive disease. capsules, p. 62 ? Are the bacteria pictured here Gram-positive or Gram-negative?
498 Chapter 21 Respiratory System Infections

Pathogenesis Treatment and Prevention

When encapsulated pneumococci are inhaled into the alveoli, Most pneumococcal infections can be cured with penicillin or
they can multiply rapidly and cause an inflammatory response. erythromycin if they are given early in the illness. However, strains
This response often affects nerve endings in the pleura, causing of pneumococci resistant to one or more antibiotics are becoming
pain, a condition called pleurisy. The bacteria are resistant to increasingly common. S. pneumoniae antibiotic resistance, p. 473
phagocytosis because their capsule interferes with the action of A vaccine is available that gives immunity to 23 pneumococ-
the complement system component C3b, an important opsonin. cus strains that cause over 90% of serious pneumococcal disease.
Pneumococcal surface protein (PspA) also interferes with the A conjugate vaccine against 13 strains is available for infants and
action of C3b. The bacteria produce pneumolysin, a membrane- children. Table  21.7 describes some features of pneumococcal
damaging toxin that destroys ciliated epithelium. The inflamma- pneumonia. conjugate vaccine, p. 424
tory response leads to an accumulation of serum and phagocytic
cells in the lung alveoli, causing breathing difficulty. This fluid
can be seen as abnormal shadows on chest X-ray films of patients Klebsiella Pneumonia
(figure  21.12). Sputum coughed from the lungs increases in Enterobacteria such as Klebsiella sp. and other Gram-negative
amount and contains pus, blood, and many pneumococci. C3b, rods can cause pneumonia, especially if host defenses are
p. 344 pleurisy, p. 486 membrane-damaging toxin, p. 392 impaired. Klebsiella sp. are common hospital-acquired pathogens
Pneumococci may enter the bloodstream from the inflamed and cause most of the deaths from healthcare-associated infec-
lungs causing three complications that are often fatal: sepsis tions. enterobacteria, p. 265 healthcare-associated infections, p. 449
(a symptomatic infection of the bloodstream); endocarditis (an
infection of the heart valves); and meningitis (an infection of the Signs and Symptoms
membranes covering the brain and spinal cord). People who do The general signs and symptoms of Klebsiella pneumonia—
not develop complications usually produce enough specific anti- cough, fever, and chest pain—are indistinguishable from those
capsular antibodies within about a week to allow phagocytosis of pneumococcal pneumonia. Klebsiella pneumonia patients
and destruction of the pneumococci. Complete recovery usually typically have repeated chills, however, and they produce thick,
results. Most pneumococcal strains do not destroy lung tissue. bloody gelatinous sputum that resembles red current jelly.
sepsis, p. 674 meningitis, p. 643 endocarditis, p. 672

Causative Agent
Epidemiology
Several species of Klebsiella cause pneumonia, but Klebsiella
Up to 30% of healthy people carry encapsulated pneumococci
pneumoniae is the best known. It is a Gram-negative rod with a
in their throat. These bacteria seldom reach the lung because
large capsule that produces big, noticeably mucoid colonies when
the mucociliary escalator effectively removes them. The risk of
grown on agar (figure 21.13).
pneumococcal pneumonia rises dramatically, however, when this
defense mechanism is impaired, as it is with alcohol and narcotic
use, and with viral respiratory infections such as influenza. There Pathogenesis
is also an increased risk of the disease with underlying heart or Klebsiella pneumoniae is contracted through inhalation, by person-
lung disease, diabetes, and cancer, and with age over 50. to-person contact, or from medical equipment such as ventilators.
Organisms first colonize the throat and gain access to the lung
via inhaled air or mucus. Specific adhesins aid colonization. The
capsule is an essential virulence factor, probably functioning like
the pneumococcal capsule in interfering with the action of comple-
ment system component C3b. Unlike Streptococcus pneumoniae,
K. pneumoniae causes tissue death and rapid formation of lung
abscesses. Therefore, even with effective antibacterial medication,
the lung can be permanently damaged and the patient may die.
The infection often enters the bloodstream, causing abscesses
in other tissues such as the liver and brain, and septic shock.
septic shock, p. 674 abscess, p. 550 complement C3b, p. 344

Epidemiology
Klebsiella sp. are widespread in nature. In humans, they can colo-
(a) (b) nize the skin, throat. and gastrointestinal tract, where they form
FIGURE 21.12 Chest X-Ray Appearance in Pneumococcal
part of the normal microbiota in some people. Typically, individu-
Pneumonia (a) Pneumonia. The left lung (right side of figure) als who contract Klebsiella pneumonia are very old, very young,
appears white because the alveoli are filled with fluid. (b) Normal or have a compromised immune system (such as alcoholics, or
X-ray film after recovery. those in a hospital or other institutional setting). The strains that
? Why does the sputum of a pneumonia patient quickly become circulate in hospitals and nursing homes are frequently resistant to
pinkish or rust-colored? antimicrobial medications and are increasingly multidrug-resistant.
 Part IV Infectious Diseases 499

TABLE 21.7 Pneumococcal, Klebsiella, and Mycoplasmal Pneumonias Compared

Pneumococcal Pneumonia Klebsiella Pneumonia Mycoplasmal Pneumonia

Signs and Cough, fever, single shaking chill, Chills, fever, cough, chest pain, and Gradual onset of cough, fever,
symptoms rust-colored sputum from degraded grossly bloody, mucoid sputum sputum production, headache,
blood, shortness of breath, chest pain fatigue, and muscle aches
Incubation 1 to 3 days 1 to 3 days 2 to 3 weeks
period
Causative agent The pneumococcus, Streptococcus Klebsiella pneumoniae, an Mycoplasma pneumoniae; lacks
pneumoniae; encapsulated strains enterobacterium cell wall
Pathogenesis Inhalation of encapsulated Aspiration of colonized mucus Cells attach to specific receptors
pneumococci; colonization of the droplets from the throat. on the respiratory epithelium;
alveoli incites inflammatory response; Destruction of lung tissue and inhibition of ciliary motion and
plasma, blood, and inflammatory abscess formation common; destruction of cells follow.
cells fill the alveoli; pain results from infection spreads via blood to other
involvement of nerve endings. body tissues.
Epidemiology High carrier rates for S. pneumoniae. Often resistant to antibiotics, and Inhalation of infected droplets;
Risk of pneumonia increased with colonize individuals who are taking mild infections common and foster
conditions such as alcoholism, them. Klebsiella sp. and other spread of the disease.
narcotic use, chronic lung disease, Gram-negative rods are common
and viral infections that impair causes of fatal healthcare-associated
the mucociliary escalator. Other pneumonias.
predisposing factors are chronic heart
disease, diabetes, and cancer.
Treatment and Treatment with penicillin, Treated with a cephalosporin with Treated with tetracycline or
prevention erythromycin, and others. Capsular an aminoglycoside. No vaccine erythromycin. No vaccine
vaccine available contains 23 capsular available. available; avoiding of crowding
antigens; conjugate vaccine for in schools and military facilities
infants. advisable.

Treatment and Prevention Treatment of these infections is becoming increasingly

Klebsiella pneumonia is treated with antibiotics. Drug sensitivity challenging. Klebsiella species commonly produce a plasmid-
tests must be carried out to determine which medications should encoded β-lactamase, an enzyme that makes the bacteria
be used. In seriously ill patients, immediate combination antibiotic resistant to all β-lactam medications (such as the penicillins).
therapy is given, often followed by a cephalosporin. Surgery may Many strains now also produce an extended-spectrum lac-
be required to drain abscesses. tamase (ESBL), which makes them resistant to many of the

Neutrophil

K. pneumoniae
cells

(a) 10 μm (b)
FIGURE 21.13 Klebsiella pneumoniae (a) In sputum from a pneumonia patient. (b) Klebsiella colonies. Notice the mucoid nature
of the colonies.
? Why is Klebsiella pneumonia often fatal?
500 Chapter 21 Respiratory System Infections

cephalosporins as well. There are very few antibiotic treatment M. pneumoniae Cilia Attachment site
choices available in these cases. Medications such as chloram-
phenicol and gentamicin are used to treat these infections with
some success, but they can have serious adverse side effects.
The case fatality rate even with treatment is as high as 50%,
and patients tend to die more quickly than other pneumonia
patients. plasmid, p. 208 β-lactamase, p. 464
There are no specific preventive measures such as vaccination
for Klebsiella pneumonia. To prevent spread between patients,
healthcare workers must follow infection control measures such
as wearing gloves and gowns when in a room with a Klebsiella
patient, and washing hands. Disinfection of the environment, use
of sterile respiratory equipment, and use of antimicrobial medica-
tions only when necessary help control the development of resis-
tance of organisms in hospitals. See table 21.7 for a description of
the main features of this disease. 0.5 μm

FIGURE 21.14 Mycoplasma pneumoniae Infecting

Respiratory Epithelium (TEM) The tips of the Mycoplasma cells
Mycoplasmal Pneumonia adjacent to the host epithelium have a distinctive appearance—they
(“Walking Pneumonia”) are probably specialized for attachment.

Mycoplasmal pneumonia is the leading kind of pneumonia in ? Why is penicillin ineffective in treating mycoplasmal pneumonia?
college students and is also common among military recruits.
The disease is generally mild (as reflected by its popular name
“walking pneumonia”) and seldom requires hospitalization.
Treatment and Prevention
Signs and Symptoms Mycoplasma pneumoniae lacks a cell wall, so antibiotics that act
The onset of mycoplasmal pneumonia is typically gradual. The against bacterial cell wall synthesis are not effective. Tetracycline
first symptoms are fever, headache, muscle pain, and fatigue. and erythromycin shorten the illness if given early, but these
After several days, a dry cough begins, but mucoid sputum may be medications are only bacteriostatic. bacteriostatic, p. 108
produced later. About 15% of cases also have otitis media. No practical preventive measures exist for mycoplasmal
pneumonia, except avoiding crowding in schools and military
Causative Agent facilities. See table 21.7 for a description of the main features of
Mycoplasmal pneumonia is caused by Mycoplasma pneumoniae, this disease.
a small, easily deformed bacterium that has no cell wall (see
figure 3.34). It grows slowly and is aerobic. Like other mycoplas- Pertussis (“Whooping Cough”)
mas, M. pneumoniae colonies on agar look like fried eggs (see
figure 11.26). mycoplasmas, pp. 62, 276
Whooping cough is the common name for pertussis, a
vaccination-preventable disease. It is still endemic in many coun-
tries, including industrialized nations such as the United States.
Pathogenesis Worldwide, the disease causes up to half a million deaths yearly.
Only a few inhaled M. pneumoniae cells are necessary to start
an infection. The organisms attach to specific receptors on the Signs and Symptoms
respiratory epithelium, interfere with ciliary action, and cause Pertussis is characterized with three stages:
the ciliated cells to slough off (figure 21.14). An inflammatory
response characterized by accumulation of lymphocytes and ■ Catarrhal stage (meaning inflammation of the mucous mem-
macrophages causes the walls of the bronchial tubes and alveoli branes). This typically begins with 1 to 2 weeks of signs and
to thicken. symptoms that resemble an upper respiratory tract infection
including runny nose, sneezing, low fever, and mild cough.
Epidemiology ■ Paroxysmal stage (meaning repeated sudden attacks). This
Mycoplasma pneumoniae is spread by aerosolized droplets of is characterized by frequent bursts of violent, uncontrollable
respiratory secretions. The organisms are shed in these secre- coughing. The cough is dry but is severe enough to rupture
tions for a long time period, ranging from about 1 week before small blood vessels in the eyes, and to cause the tongue to
symptoms begin to many weeks afterward, thereby increasing the protrude and the neck veins to stand out. The coughing spasm
likelihood of transmission. Mycoplasmal pneumonia accounts for is followed by forceful attempts to inhale. The inspired air is
about one-fifth of bacterial pneumonias, and has a peak incidence gasped in, causing the characteristic “whoop” of this disease.
in young people. Immunity after recovery is not permanent, and Vomiting and seizures can occur during this stage and the
repeat attacks have occurred within 5 years. patient may also become cyanotic (blue from lack of O2).
 Part IV Infectious Diseases 501

■ Convalescent stage. During this stage the person is no longer Pertussis toxin (PTx)
contagious. The coughing attacks gradually become less fre-
quent, and the person slowly recovers. B B
B B B
A
Causative Agent
Receptor
Whooping cough is caused by Bordetella pertussis, a tiny, encap-
sulated, strictly aerobic, Gram-negative rod (figure 21.15). These G
organisms are sensitive to drying and sunlight, and die quickly 1 The B subunits of PTx bind
Host cell to receptors on the host cell
outside the host. surface. The A subunit is
surface Regulatory
transferred into the host cell,
protein
Pathogenesis becoming an activated
enzyme on entry.
When Bordetella pertussis is inhaled, it attaches specifically to
ciliated cells of the respiratory epithelium. Attachment is aided by
two colonization factors: (1) filamentous hemagglutinin (FHA),
a pilus that extends from the bacterial surface; and (2) pertussis
toxin (PTx), a protein that functions as an adhesin, as well as B B
having toxic effects (described later). Both factors are needed B B B
for colonization. The areas colonized by B. pertussis include the
nasopharynx, trachea, bronchi, and bronchioles. The organisms
grow in dense masses on the epithelial surface, but generally do
G
not invade the cells. Instead, they release three toxins (described A
2 Regulatory protein G is
next) that play critical roles in the disease process.
inactivated by enzyme A,
Pertussis toxin (PTx) is an A-B exotoxin (figure  21.16). causing an increase in cAMP Activated
The B subunit attaches specifically to receptors on the host production. enzyme A
cell surface, allowing the A subunit to move through the cyto-
plasmic membrane of the host cell. As it does so, it activates a
FIGURE 21.16 Mode of Action of Pertussis Toxin (PTx)
membrane-bound regulatory protein that controls the production
of cyclic adenosine monophosphate (cAMP), leading to increased ? What is the result of increased cAMP production?
production of this molecule. High levels of cAMP interfere with
cell signaling pathways, resulting in marked increase in mucus
output, decreased killing ability of phagocytes, massive release of lymphocytes into the bloodstream, ineffectiveness of natural killer
cells, and low blood sugar. A-B toxins, p. 392 cAMP, p. 587
Adenylate cyclase is both a membrane-damaging toxin and an
enzyme. This toxin reduces phagocytosis by causing lysis of accu-
Ciliated Cluster of
mulating leukocytes. Inside the cell, it also catalyzes the reaction
epithelial cell B. pertussis that converts ATP to cAMP.
Tracheal cytotoxin is a fragment of peptidoglycan that
B.  pertussis releases during growth, and it causes host cells to
release a fever-inducing cytokine (interleukin-1; IL-1). It is also
toxic to ciliated epithelial cells, causing them to die and slough
off, resulting in a rapid decline in ciliary action. The combination
of increased mucus production and decreased ciliary action results
in the cough of pertussis because only the cough reflex remains for
clearing chest secretions. Some of the bronchioles become com-
pletely obstructed by mucus, resulting in small areas of collapsed
lung. Spasms or partial mucus plugging in other bronchioles let
air enter but not escape, causing hyperinflation. Pneumonia due to
B.  pertussis or, more commonly, secondary bacterial infection is
the main cause of death. interleukin-1, p. 341

Epidemiology
Pertussis is highly contagious, spread via respiratory secre-
10 μm tions suspended in air. Patients are most infectious during the
FIGURE 21.15 Bordetella pertussis Fluorescent antibody stain catarrhal stage. Once the paroxysmal period begins, the numbers
of respiratory secretions from a person with whooping cough. of expelled organisms decreases substantially. Pertussis is classi-
? Why are the B. pertussis cells clustered at one end of the cally a disease of infants. However, it occurs in a milder form in
ciliated cell? older children and adults, and may be overlooked as a persistent
502 Chapter 21 Respiratory System Infections

cold, asthma, or bronchitis, thus fostering transmission. Despite 20

Reported cases per 100,000 population

the successful vaccine, the reported incidence of the disease is 18
steadily increasing. Possible factors contributing to this trend
16
include better surveillance and diagnostic techniques, suboptimal
14
vaccination (especially in developing nations), decreasing immu-
nity, and development of B. pertussis strains for which the current 12
vaccine is not effective. In 2010, an outbreak of pertussis occurred 10
in California. At least ten infants died of the disease, and health 8
authorities reported an epidemic. 6
4
Treatment and Prevention 2
Erythromycin reduces the duration of pertussis symptoms if 0
given during the catarrhal stage, and the antibiotic usually elimi- 1978 1983 1988 1993 1998 2003 2009
nates B.  pertussis from the respiratory secretions. Azithromycin Year
and trimethoprim-sulfamethoxazole are effective alternatives.
Antibiotics are ineffective in the paroxysmal stage of pertussis. FIGURE 21.17 Incidence of Tuberculosis, United States,
Pertussis is effectively prevented with a vaccine. The original 1978–2009
vaccine, composed of whole B. pertussis cells, produced a dra- ? What caused the rise in TB incidence between 1988 and 1994?
matic decrease in pertussis cases, but sometimes caused severe
side effects. It has now been replaced with a newer acellular per-
tussis vaccine (aP), that includes only part of the bacterium instead epidemic and increasing prevalence of drug-resistant strains of the
of whole cells. It is given in combination with diphtheria and teta- causative agent (figure  21.17). To respond to the problem, the
nus toxoids—a grouping referred to as the DTaP vaccine—at 2, 4, CDC developed a Strategic Plan for the Elimination of Tuberculosis
6, and 15 to 18 months, and again at 4 years old. A booster with a in the United States, published in 1989. The plan depends largely
decreased dose of pertussis antigen (Tdap) is given every 10 years on increased efforts in identifying and treating cases among the
after that. The main features of pertussis are shown in table 21.8. high-risk groups, particularly poor people, people with AIDS,
acellular vaccines, p. 423 prisoners, and immigrants from countries with high rates of TB.
By 1993, the incidence began to decrease again and by 2009, only
3.8 cases per 100,000 population were recorded.
Tuberculosis (“TB”)
MicroByte
Tuberculosis (TB) was once a very common disease but the
It is estimated that one-third of the global population is infected with
number of cases gradually declined in industrialized countries as Mycobacterium tuberculosis and almost 2 million die of tuberculosis
living standards improved. In 1985, however, the incidence of TB annually.
began to rise again, the trend associated with the expanding AIDS

TABLE 21.8 Pertussis

M. tuberculosis
Signs and Runny nose followed after a number
symptoms of days by spasms of violent coughing;
vomiting and possible convulsions
Incubation 7 to 21 days
period
Causative agent Bordetella pertussis, a tiny Gram-negative
rod
Pathogenesis Colonization of the surfaces of the upper
respiratory tract and tracheobronchial
system; ciliary action slowed; toxins
released by B. pertussis cause death of
epithelial cells and increased cAMP; fever,
excessive mucus output, and a rise in the
number of lymphocytes in the bloodstream
result.
Epidemiology Inhalation of infected droplets; older
children and adults have mild symptoms.
Treatment and Erythromycin, somewhat effective if given
prevention before coughing spasms start, eliminates
B. pertussis. Acellular vaccine (DTaP), for FIGURE 21.18 Acid-fast Stain of Myobacterium tuberculosis
immunization of infants and children.
? Why is M. tuberculosis so resistant to drying?
 Part IV Infectious Diseases 503

Signs and Symptoms and multiply within the cytoplasm of the macrophages (figure
The initial infection with Mycobacterium tuberculosis typi- 21.19).
cally results in an asymptomatic lung infection. The immune The multiplying bacilli trigger an inflammatory response,
response generally controls this primary infection but is not able recruiting more macrophages to the site, thereby providing
to eliminate it entirely—the person is left healthy but with M. tuberculosis with additional host cells in which to multiply.
a latent infection, known as latent tuberculosis infection Some of the macrophages fuse together to form giant multi-
(LTBI). Much later in life, the person may develop active nucleated cells. Others are induced by the bacteria to accumulate
tuberculosis disease (ATBD), a chronic illness characterized large numbers of oil droplets, becoming foamy macrophages.
by slight fever, progressive weight loss, night sweating, and The lipids in foamy macrophages are thought to help the bacteria
persistent cough, often produc-
ing blood-streaked sputum. Some
people, especially children or Mycobacteria
1
those with compromised immune
Blood vessel
systems, may develop ATBD on Alveolar macrophages
primary infection. latent infection, ingest mycobacteria.
p. 383 Infected alveolar
macrophage Interior of alveolus

Causative Agent 2 Early tubercle

Lymphocyte
Tuberculosis is caused by Bacteria survive and
Mycobacterium tuberculosis, com- Foamy multiply in macrophages.
Infected
Additional macrophages
monly called the tubercle bacillus. macrophage macrophage
and lymphocytes are
The organism is a slender, acid- recruited to site. Foamy
fast, rod-shaped bacterium (figure  macrophages develop.
Blood
Macrophage
21.18). It is a strict aerobe that vessel
grows very slowly, with a genera-
tion time of over 16 hours. This
slow growth makes it difficult to 3
diagnose TB quickly. The organism Lymphocyte
Fibrous layer
has an unusual cell wall that con- Infected
Foamy macrophage Fibrous capsule surrounds
tains a large amount of complex macrophage macrophages, excluding
glycolipids called mycolic acids— lymphocytes.
these make it unusually resistant to Macrophage
drying, disinfectants, and strong
acids and alkali, although it is easily
killed by pasteurization. The mycolic
acids are also largely responsible for
its acid-fast staining. M. tuberculosis 4

primarily infects the lung but can

also cause disease in many other Foamy Caseum Infected macrophages die,
tissues, including bones, kidneys, macrophage releasing mycobacteria
and creating caseous
joints, and the central nervous sys- necrosis.
tem. pasteurization, p. 112 lipids, Macrophage
p. 33 acid-fast staining, p. 48

Pathogenesis 5 Free mycobacteria

When airborne M. tuberculosis Bronchiole
cells from a person who has active Tubercle ruptures,
Tubercle
TB disease are inhaled, they may releasing live mycobacteria
enter the lungs. Alveolar macro- into the airway.
phages quickly engulf the bacteria
but are unable to destroy them
because the mycolic acids in the
bacterial cell wall prevent fusion
of the phagosome with lysosomes. FIGURE 21.19 Pathogenesis of Tuberculosis
The bacteria leave the phagosome ? Why is prolonged treatment necessary for tuberculosis?
504 Chapter 21 Respiratory System Infections

survive within the cells. Lymphocytes collect around the macro- by conditions in the tubercle, including low pH and low available
phages, walling off the infected area from the surrounding tissue. O2. The bacteria remain in this state for many years, causing a
This localized collection of inflammatory cells—a granuloma—is latent TB infection (LTBI). Individuals with LTBI are asymp-
the body’s characteristic response to microorganisms and other tomatic and non-infectious. In many cases, the infection resolves.
foreign substances that resist destruction and removal by phago- granuloma, p. 348
cytosis. The granulomas of tuberculosis are called tubercles Active TB disease results if the inflammatory response can-
(figure 21.20; see also figure 21.19). Within the granuloma, effec- not contain or destroy the mycobacteria. This can occur during
tor helper T cells release cytokines that activate macrophages to primary infection but can also happen in a person with LTBI—the
destroy the bacteria infecting them. At this time, a fibrous layer infection reactivates (reactivation TB) if the person’s immunity
forms around the macrophages, keeping the lymphocytes outside becomes impaired by stress, advanced age, or disease such as
of the tubercle. This fibrous tissue can be seen on X rays as Ghon AIDS. Within the tubercle, macrophages containing mycobac-
foci. If the adjacent lymph nodes are involved, the focus is called teria die, releasing bacteria, enzymes, and cytokines. An area
a Ghon complex. Some of the mycobacteria in the Ghon foci of necrosis is formed in the center of the tubercle—this has the
and complexes survive, but they are prevented from multiplying texture of soft white cheese and is referred to as caseous necrosis.
It is thought that foamy macrophages play an important role in
necrosis formation and that the caseum contains lipids from these
cells. The tubercle then ruptures, releasing the bacteria and dead
material into the airways. This causes a large lung defect called
a tuberculous cavity that spreads the bacteria to other parts of the
lung. Lung cavities characteristically persist, slowly enlarging for
months or years and shedding bacterial cells into the bronchi. The
Cavities organisms can then be transmitted to other people by coughing
and spitting.
Other systems that can be affected by reactivation TB
include the pleura and pericardium, lymph nodes, kidneys, bones
(typically the lumbar and thoracic vertebrae which break down,
causing Pott’s disease), joints, and central nervous system. In a
condition called miliary tuberculosis, tiny tubercles are found in
multiple organs throughout the body.

Epidemiology
(a) An estimated 15 million Americans have LTBI, but the vast
majority of these will never develop active TB disease (ATBD)—
only 5% to 10% of latent infections will later reactivate, resulting
in progression to ATBD. LTBI rates are highest among non-
whites and elderly poor people. Foreign-born U.S. residents have
much higher incidence of LTBI than those born in the United
States. Transmission of M. tuberculosis occurs almost entirely by
the respiratory route; 10 or fewer inhaled organisms are enough
to cause infection. Factors important in transmission include the
frequency of coughing, the adequacy of ventilation (transmis-
Boundary of sion is unlikely to occur outdoors), and the degree of crowding.
necrotic area Immunodeficiency increases activation of M. tuberculosis in those
with LTBI, a significant problem in AIDS patients.
The tuberculin skin test (TST), also known as the Mantoux
(pronounced man-too) test, is an extremely important tool
for studying the epidemiology of the disease and in detecting
(b) 1 mm
those who are infected with M. tuberculosis. The test is carried
out by injecting into the skin a small amount of a sterile fluid
FIGURE 21.20 Stained Lung Tissue Showing a Tubercle
called purified protein derivative (PPD), derived from cultures
(a) Chest X ray of a person with TB. (b) Lung tissue showing tubercle.
The dark dots around the outer portion of the picture are nuclei of of M. tuberculosis. People who are infected with the bacte-
lung tissue and inflammatory cells. In the center of the photograph, rium develop redness and a firm swelling at the injection site,
most of the nuclei have disappeared because the cells are dead and reaching a peak intensity after 48 to 72 hours (figure  21.21).
the tissue has begun to liquefy. This reaction is due to the accumulation of macrophages and
? What is a tubercle? T lymphocytes at the injection site, a manifestation of delayed
 Part IV Infectious Diseases 505

regime. What commonly happens is that once a person feels

better and the symptoms of active TB disease disappear, he or
she becomes careless about continuing to take the prescribed
medications. To combat this problem, DOTS (directly observed
therapy short-course) may be used—healthcare workers that
supply the medications can watch the patients swallow the pre-
scribed tablets.
Despite the success of DOTS, the problem of drug-
resistant M. tuberculosis strains has nevertheless reached
alarming proportions in some areas. During the 1990s, multidrug-
resistant TB (MDR-TB) became an increasing problem. These
strains resist rifampin and isoniazid—the two most effective
first-line drugs available—and therefore must be treated with
FIGURE 21.21 Tuberculin Skin Test This positive test is caused less effective, more toxic (causing liver damage), and more
by delayed hypersensitivity to Myobacterium tuberculosis antigens expensive second-line medications. By the end of the decade,
injected into the skin. extensively drug-resistant TB (XDR-TB) strains that resist
? What does a positive tuberculin skin test indicate? both first-line and many of the second-line drugs evolved. These
strains now threaten tuberculosis control efforts around the
world. Fortunately, promising new anti-tuberculosis medications
representing at least five different chemical families are being
hypersensitivity to the tubercle bacillus. A positive reaction to developed. One entirely new and highly active medication—
the test (a skin reaction more than 10 mm in diameter) does not a diarylquinoline (DARQ)—is among those under evaluation.
MDR-TB, p. 473 XDR-TB, p. 473
mean that the person has ATBD, only that the person has been
infected by M. tuberculosis at some time in the past; they may In the United States, TB prevention involves identifying
have either LTBI or ATBD. They may also have received a TB unsuspected cases using skin tests and lung X rays. Individuals
immunization. delayed hypersensitivity, p. 409
with active disease are then treated, thereby interrupting the
Beside the TST, blood tests are available for diagnosing spread of the causative agent. People who have LTBI are also
M. tuberculosis infection. Interferon gamma–release assays (IGRAs) treated, reducing the risk of developing ATBD later in life.
detect interferon gamma released in response to M. tuberculosis. The preferred treatment for LTBI is isoniazid for 9 months.
IGRAs generally give results similar to TST but more quickly and Treatment is particularly recommended for high-risk individu-
with greater specificity. Nucleic acid amplification methods are als such as those with underlying conditions (including those
fast, sensitive, and accurate, and are used on clinical specimens. with HIV infection, drug abuse, and diabetes), the very young,
IFN-gamma, p. 341 nucleic acid amplification tests, p. 246
the elderly, those recently exposed to ATBD, those recently
converted to a positive tuberculin skin test, and those entering
the country from regions with high prevalence of the disease.
Treatment and Prevention The National Tuberculosis Indicators Project (NTIP) col-
Treatment of tuberculosis requires multiple medications for many lects data on individual TB cases to measure the performance
months. A combination of drugs must be given because people of prevention and control measures in the United States.
with ATBD have high numbers of M. tuberculosis cells in the The CDC has also initiated the Tuberculosis Genotyping
body, so there is a strong likelihood that some of the cells will Information Management System (TB GIMS) to disseminate
have acquired resistance to a given drug by spontaneous mutation. data on suspected TB outbreaks so that they can be managed
A cell is much less likely to develop resistance to two drugs given more efficiently.
simultaneously. Treatment must be continued for months to cure Prevention and control of TB is a global challenge and many
the disease because of the long generation time of M. tuberculosis countries lack the resources to track and treat both ATBD and
and its resistance to destruction by body defenses. antibiotic LTBI. Vaccination against TB has been widely used in many
resistance, p. 471 generation time, p. 83 countries, The vaccine used, BCG (Bacille Calmette-Guérin),
The most effective and least toxic first-line medications for is a live attenuated vaccine derived from M. bovis, a cattle-
treating ATBD include rifampin (RIF), isoniazid (INH), pyra- infecting species that has little virulence in humans. Although
zinamide (PZA), and ethambutol (EMB), which are bactericidal this vaccine prevents childhood TB disease, it appears ineffec-
against actively growing organisms. Some kill metabolically inac- tive in preventing LTBI, which can later reactivate. Use of the
tive intracellular organisms as well. In the initial treatment phase vaccine is discouraged in the United States, because people who
of ATBD, all four first-line drugs are given for 2 months. After receive it usually develop a positive tuberculin skin test. By
that, INH and RIF are given in combination for another 4 to 7 causing a positive test, this vaccination eliminates an important
months, depending on the number of doses scheduled each week. way of diagnosing TB early in the disease when it can most
Strains resistant to one or more first-line medications often easily be treated. BCG is also not safe to use in severely immu-
evolve when people fail to comply with the complex treatment nocompromised patients. Several new genetically engineered
506 Chapter 21 Respiratory System Infections

TABLE 21.9 Tuberculosis

1 Airborne Mycobacterium 4 Signs and Chronic fever, weight loss, cough, sputum
3
tuberculosis cells are inhaled 5 symptoms production
and lodge in the lungs.
1 Incubation 2 to 10 weeks
2 The bacteria are phagocytized period
3
by lung macrophages and 7 5 4 Causative agent Mycobacterium tuberculosis; unusual cell wall with
multiply within them,
high lipid content
protected by lipid-containing 1
cell walls and other 2 Pathogenesis Colonization of the alveoli incites inflammatory
mechanisms. 6 response; ingestion by macrophages follows;
organisms survive ingestion and are carried to
3 Infected macrophages are
lymph nodes, lungs, and other body tissues;
carried to various parts of
tubercle bacilli multiply; granulomas form.
the body such as the kidneys, 3 5
brain, lungs, and lymph nodes; 4 Epidemiology Inhalation of airborne organisms; latent infections
release of M. tuberculosis can reactivate.
occurs.
Treatment and Treatment: two or more antitubercular medications
4 Delayed hypersensitivity prevention given simultaneously long term, such as isoniazid
develops; wherever infected (INH) and rifampin; DOTS; BCG vaccination
M. tuberculosis has lodged, an preventive but not used in the United States;
intense inflammatory reaction tuberculin (Mantoux) skin test for detection of
develops. infection, allows early therapy of cases; treatment
of all high-risk cases including young people with
5 The bacteria are surrounded by
positive tests and individuals whose skin test
macrophages and lymphocytes;
converts from negative to positive.
growth of the bacteria ceases.
6 Intense inflammatory reaction
and release of enzymes can
cause caseation necrosis and
cavity formation.
7 With uncontrolled or reactive
infection, M. tuberculosis exits
the body through the mouth
with coughing.

vaccines are being developed, many of which are currently being culture (figure  21.22). Its fastidious growth requirements and
tested in TB-endemic regions. The main features of tuberculosis the fact that the cells stain poorly in tissue partly explain why
are shown in table 21.9. this organism was undetected for so long. L. pneumophila is a
facultative intracellular parasite and survives well in freshwater
Legionnaires’ Disease
Legionnaires’ disease was unknown until 1976, when a number of
people attending an American Legion Convention in Philadelphia
developed a mysterious pneumonia that was fatal in many cases.
Months of scientific investigation eventually paid off when the
cause was discovered to be a previously unknown bacterium com-
monly present in the natural environment.

Signs and Symptoms

Legionnaires’ disease typically begins with headache, muscle
aches, high fever, confusion, and shaking chills. A dry cough
develops that later produces small amounts of sputum, sometimes
containing blood. Pleurisy can also occur. About one-fourth of the
cases also have some digestive tract symptoms such as diarrhea,
abdominal pain, and vomiting. Shortness of breath is common,
and O2 therapy is often needed. Recovery is slow, and weakness
and fatigue last for weeks. 10 μm
FIGURE 21.22 Legionella pneumophila in Lung Tissue
Causative Agent Stained with Fluorescent Antibody The bacterium does not stain
Legionnaires’ disease is caused by Legionella pneumophila, a with most of the usual microbiological stains in tissue or sputum.
Gram-negative rod that requires a special medium for laboratory ? How is L. pneumophila acquired?
 Part IV Infectious Diseases 507

amebas such as Acanthamoeba. These protozoa form cysts during

TABLE 21.10 Legionnaires’ Disease
adverse environmental conditions, allowing the bacteria within
them to survive. Legionella also persists in biofilms—if the bio- Signs and Muscle aches, headache, fever, cough,
film is disturbed, huge numbers of Legionella are released into symptoms shortness of breath, chest and abdominal
pain, diarrhea
the water.
Incubation 2 to 10 days
period
Pathogenesis
Causative agent Legionella pneumophila, a Gram-negative
Legionella pneumophila is acquired by breathing aerosolized bacterium that stains poorly in clinical
water contaminated with the organism. Healthy people are quite specimens
resistant to infection, but smokers and those with impaired host Pathogenesis Organism multiplies within phagocytes;
defenses are susceptible. released with death of the cell; necrosis of
The organisms lodge in and near the alveoli of the lung. cells lining the alveoli; inflammation and
Rather than avoiding phagocytosis by alveolar macrophages, they formation of microabscesses
promote it. One of their surface proteins, macrophage invasion Epidemiology Originates mainly from warm water
potentiator (Mip), aids entry into the macrophages. The bacterial contaminated with other microorganisms,
cells also bind complement component C3b, an opsonin. Once such as found in air-conditioning systems.
ingested by the macrophages, the bacteria survive by preventing Treatment and Treatment: erythromycin and rifampin.
phagosome-lysosome fusion. They also manipulate other events prevention Avoidance of contaminated water aerosols;
within the phagocyte, creating an environment in which they can regular cleaning and disinfection of
humidifying devices.
multiply. The host cell eventually dies, releasing bacterial cells
that can then infect other tissues. Necrosis (tissue death) of alveo-
lar cells and an inflammatory response result, causing multiple
small abscesses, pneumonia, and pleurisy. Bacteremia is often MicroAssessment 21.4
present. Fatal respiratory failure (meaning the lungs can no longer Pneumococcal pneumonia is typically acquired in the community
adequately oxygenate the blood or expel CO2) occurs in about and leads the list of pneumonias in adults requiring hospitalization.
15% of hospitalized cases. Curiously, L. pneumophila infections Pneumonia due to Klebsiella sp. and other Gram-negative rods is
remain confined to the lung in most cases. mainly hospital-acquired and leads the causes of death from healthcare-
associated infections. Mycoplasmal pneumonia usually does not
require hospitalization. Whooping cough (pertussis) is mainly a threat
Epidemiology to infants but is commonly spread by adults; childhood immunization
Legionella pneumophila is widespread in warm natural waters against the disease protects them, but immunity often does not persist
containing other microorganisms such as amebas, in which the to adulthood. Tuberculosis is a chronic disease spread from one
person to another by aerosol drops. Most Mycobacterium tuberculosis
bacteria live and multiply. The organism also survives well in
infections become latent, posing the risk of reactivation throughout
the water systems of buildings, particularly in hot water systems, life. Legionella pneumophila, the bacterium that causes Legionnaires’
where chlorine levels are generally low. Increased chlorine lev- disease, originates from water containing other microorganisms, where
els sometimes fail to decontaminate water systems, probably it can grow within protozoa. In the human lung, the bacterium readily
because the L. pneumophila cells are protected inside amebas. multiplies within macrophages.
Legionnaires’ disease cases have originated from contaminated 10. What structural feature of the S. pneumoniae cell is responsible
aerosols from large central air-conditioning systems, nebulizers, for its virulence?
sprays to freshen produce, and from showers and water faucets. 11. Outline the pathogenesis of tuberculosis.
People have even contracted this disease from car windshield 12. Why is pertussis toxin not eliminated by the mucociliary
sprays that do not contain detergent. Direct person-to-person escalator? +
spread, however, does not occur.

Treatment and Prevention 21.5 ■ Viral Infections of the

Legionnaires’ disease is treated with high doses of erythromycin, Lower Respiratory System
sometimes concurrently with rifampin. L. pneumophila produces
a β-lactamase, which makes it resistant to many penicillins and Learning Outcomes
some cephalosporins. Also, since the bacteria multiply inside 9. Describe antigenic drift and antigenic shift and discuss how they
the alveolar macrophages, the medication used must be able to affect the epidemiology of influenza.
accumulate within these cells, which β-lactam drugs do poorly. 10. Compare the distinctive characteristics of respiratory syncytial
 β-lactamase, p. 464 virus infection and hantavirus pulmonary syndrome.
Most efforts at control of Legionnaires’ disease have focused
on designing equipment to minimize the risk of infectious aerosols DNA viruses such as the adenoviruses sometimes cause serious
and on disinfecting procedures. Environmental surveillance is not pneumonias, but RNA viruses are of greater overall importance
practical because of the lack of a simple method for detecting because of the large number of people they infect and their poten-
virulent strains. The main features of Legionnaires’ disease are tial for serious outcomes. The following section covers some
given in table 21.10. diseases caused by RNA viruses.
508 Chapter 21 Respiratory System Infections

Influenza (“Flu”) the envelope are two kinds of glycoprotein spikes—hemagglutinin

antigen (HA) and neuraminidase antigen (NA)—which have a role
Influenza is a good example of the constantly changing interac-
in viral pathogenesis. The HA spikes allow the virus to recognize
tion between people and infectious agents. Antigenic changes in
and attach to specific receptors on ciliated host epithelial cells,
the influenza viruses are responsible for serious annual epidemics
initiating infection. Viruses with HA spikes cause red blood cells
of the disease—almost 20% of the world population gets infected
to stick together (hemagglutination), a useful characteristic for
with flu virus every year. There are three major influenza virus
virus identification. NA is an enzyme that plays a critical role in
types, named A, B, and C, based on differences in their protein
the release of newly formed virions from host cells. As new virions
coat. Type A, considered here, causes the most serious disease,
are made, they bud out of the host cell but remain bound to sur-
and its epidemics are widespread. Outbreaks due to type B strains
face receptors in the host membrane. NA destroys these receptors,
occur each year, but they are less extensive and the disease is
allowing the virions to leave the infected host cell, and aiding the
not as severe. Type C strains are of relatively little importance.
spread of the virus to uninfected host cells.
Influenza viruses do not cause “stomach flu.”
There are different subtypes of influenza A viruses, character-
ized by antigenically distinct HA and NA spikes. The subtypes are
Signs and Symptoms
given numbers according to these variations—H1, H2, N1, N2 and
After a short incubation period averaging 2 days, influenza typically so on. For example, the “avian flu” epidemic of 1997 was caused
begins with headache, fever, sore throat, and muscle pain, peaking by influenza virus H5N1, whereas the “swine flu” epidemic of
in 6 to 12 hours. A dry cough develops and worsens over a few days. 2009 was caused by influenza virus H1N1. There are 16 HA and
These acute symptoms usually go away within a week, leaving the 9 NA subtypes, but only H1, 2 and 3, and N1 and 2 spread among
patient with a lingering cough, fatigue, and generalized weakness humans (see Perspective 21.2).
for additional days or weeks. Infection by influenza viruses is occa-
sionally associated with Reye’s syndrome, a complication linked to Pathogenesis
certain other viral infections as well. Reye’s syndrome, p. 535
Individuals acquire influenza by inhaling aerosolized respira-
tory secretions from a person who has the disease. They can also
Causative Agents
contract the virus from fomites such as doorknobs, banknotes,
Influenza A virus belongs to the orthomyxovirus family. It has and household items, accidentally transferring it into the eyes or
eight segments of single-stranded RNA, which are enclosed in nasal passages by touch. The virions attach by their HA spikes to
a protein capsid. The capsid is surrounded by a lipid envelope specific receptors on ciliated respiratory epithelial cells, and enter
derived from the host cell membrane the cell by endocytosis. Viral RNA is released into the host cell
(figure  21.23). Embedded in cytoplasm following fusion with the endosomal membrane. Host
cell protein and nucleic acid synthesis stop, and rapid synthesis
of viral RNA and proteins begins. Regions of the host cell mem-
brane become embedded with virally encoded HA and NA glyco-
Lipid envelope
proteins. Within 6 hours, mature virions bud from the host cell,
acquiring host cell-derived membrane containing these HA and
Nucleoprotein NA spikes as they do so. The virus spreads rapidly to nearby cells,
including mucus-secreting cells and cells of the alveoli. Infected
Hemagglutinin cells die and slough off, thus destroying the mucociliary escalator.
(HA) The damage to this important first line of defense makes the per-
son susceptible to secondary respiratory infections. The immune
Neuraminidase response quickly controls the influenza virus infection in most
(NA) cases, although complete recovery of the respiratory epithelium
may take 2 months or more.

RNA segments Epidemiology

Matrix protein Usually, only a small percentage of people with influenza die, but
many people fall ill during an epidemic so the total number of
deaths is high. Although influenza virus infection alone can kill
otherwise healthy people, most deaths are due to bacterial second-
ary infections such as pneumonia. People are predisposed to these
infections because of the virally induced damage to the respiratory
epithelium. In fact, H. influenzae got its name because it was often
FIGURE 21.23 Structure of Influenza Virus found in the lungs of people who died after having influenza, lead-
? What is the role of the HA and NA spikes? ing to the incorrect conclusion that it was the cause of the disease.
 Part IV Infectious Diseases 509

PERSPECTIVE 21.2
What to Do About Bird Flu
The known antigenic types of influenza A in 1997. Moreover, the virus has spread rap- highly likely that other avian strains will do so
are 16 different hemagglutinins, designated idly from Asia to the Middle East, Europe, sometime in the future. We have a historical
H 1–16, and nine neuraminidases, N types and Africa (figure  1), devastating flocks of example in the 1918–1919 “Spanish flu” that
1–9. All of these are represented among the domestic fowl and causing several hundreds killed 40 to 100 million people. This pandemic
influenza A viruses infecting wild waterfowl. of deaths among humans. Fortunately, so far was caused by an avian virus.
Generally, only H types 1–3, and N types 1 there have been only a few instances that sug- Today, we are using some important tools
and 2 viruses have been important in human gest person-to-person spread of the disease. to defend ourselves that were not available in
disease. Occasionally, however, avian viruses The H5N1 avian virus concerns us 1918. Rapid viral diagnosis is now possible,
infect humans and cause illness. For example, because it has two of the three characteris- and global communication can inform us
there were 123 known human cases caused tics required of a pandemic influenza virus: promptly of outbreaks anywhere in the world
by avian viruses between 1996 and 2004, (1) It is infectious for humans who have no so we can take defensive measures. We can
caused by five avian strains: H7N7, H5N1, herd immunity; and (2) it causes severe disease also make and stockpile influenza vaccines,
H9N2, H7N2, and H7N3. There was a high in humans. The virus does not yet meet the as well as vaccines against bacterial secondary
mortality among the 29 cases caused by the third requirement—easy spread from person invaders such as Streptococcus pneumoniae
H5N1 strains (seven died), whereas only one to person, but it might acquire this ability and Haemophilus influenzae. We can also
death occurred among the remaining 103 cases through mutation or antigenic shift. herd place antiviral and antibacterial medications
due to the other four avian viruses. The high immunity, p. 421 at strategic locations. A great deal depends on
virulence of the H5N1 strain has been appar- The H5N1 virus may never become a international cooperation.
ent since the first H5N1 outbreak occurred pandemic strain, but even if it does not, it is

United Kingdom
of Great Britain Sweden Russian
and Northern Ireland Federation
Denmark
Netherlands
Poland Czech Republic
Germany
Slovakia
Austria Hungary Ukraine Kazakhstan
France
Romania Mongolia
Switzerland Italy Serbia
Georgia
Bulgaria Azerbaijan
Spain Croatia Republic
Greece Turkey
of Korea

Albania China Japan

Islamic Afghanistan
Iraq Republic
of Iran
Pakistan

Egypt Myanmar
Saudi
Arabia
India Thailand Lao People’s
Burkina Niger Bangladesh Democratic Republic
Faso
Vietnam
Sudan Cambodia
Nigeria
Malaysia
Cameroon

Cote d’Ivore
Indonesia

Areas reporting occurrence in poultry

Areas reporting occurrence only in wild birds

FIGURE 1 Areas Reporting H5N1 Avian Influenza in Poultry and Wild Birds Confirmed reports since 2003, last updated
September 2007.
510 Chapter 21 Respiratory System Infections

It actually causes pneumonia, among other diseases. secondary Two types of variation occur: antigenic drift and antigenic shift
infection, p. 382 (figure 21.24):
Influenza epidemics occur every year. Pandemics occur
periodically over the years, marked by rapid spread of the ■ Antigenic drift. This is caused by minor mutations in the
viruses around the globe and higher than normal morbidity. genes that code for the HA and NA antigens and is respon-
Several factors are involved in the spread of influenza viruses, sible for the yearly occurrence of influenza outbreaks, called
but major attention has focused on their antigenic changeability. seasonal influenza. The mutations happen during normal viral

Antigenic Drift (Seasonal Influenza)

H antigen

N antigen
Mutation #2
Mutation #1
Genome
segments

Mutation #1

Mutation #1 Mutation #2

Mutation #2

(a)

Antigenic Shift (Pandemic Influenza)

Viral RNA replication

and reassortment

New variant of a
human influenza
with duck N spike

Nucleus

Pig lung cell

(b)
FIGURE 21.24 Influenza Virus: Antigenic Drift and Antigenic Shift With drift, repeated mutations cause a gradual change in the HA
and/or NA spikes, so that antibody against the original virus becomes progressively less effective. With shift, there is a sudden major change in the
spikes because the virus acquires a new genome segment.
? Why can antigenic shifts cause pandemics?
 Part IV Infectious Diseases 511

replication and often cause a change in only a single amino MicroByte

acid in the HA or NA spikes. They occur frequently, however, Outbreaks of influenza occur every year in the United States and are
and are enough to make immunity developed to virus strains associated with an estimated 10,000 to 40,000 deaths.
of previous years less effective, ensuring a continual supply of
susceptible hosts within which the virus can multiply. Strains
Treatment and Prevention
that arise because of antigenic drift are given names indi-
cating the year and location they were isolated. For example, Like all viral infections, antibiotic treatment is not effective for
a strain referred to as A/Texas/77 (H3N2) is an influenza influenza. Medications such as amantadine (Symmetrel) and
A strain isolated in 1977 in Texas, whereas A/Bangkok/79 rimantadine (Flumadine) have been used for short-term preven-
(H3N2) occurred in Bangkok in 1979. The Bankok/79 strain tion of influenza A disease when vaccination is not an option
has minor HA spike mutations that distinguish it from the or while waiting for the protection of vaccination to take effect.
Texas/77 strain. The antibody produced by people who have However, they are not currently recommended because many
recovered from A/Texas/77 (H3N2) is only partially effective circulating viruses are resistant to them. Zanamivir (Relenza) and
against the A/Bangkok/79 (H3N2). Thus, the newer Bangkok oseltamivir (Tamiflu) are neuraminidase inhibitors, active against
strain might be able to spread and cause a minor epidemic in both A and B viruses. These medicines are generally used together
people previously exposed to the Texas strain. with vaccination to protect exposed individuals until they can
develop immunity. neuraminidase inhibitors, p. 476
■ Antigenic shift. This is an uncommon but more dramatic Prevention is by vaccine, but there is no vaccine that pro-
change that occurs as a result of viral genome reassortment duces lifelong immunity to influenza, because of antigenic drift—
and is the cause of pandemic influenza. Recall that the every year a slightly new strain of virus emerges, against which
influenza virus genome is segmented, meaning that viral immunity must be generated. Inactivated vaccines are developed
proteins are encoded on eight different RNA segments rather annually against the three most important influenza strains in cir-
than being encoded on one long molecule. Because of this culation at that time. It takes 6 to 9 months from the appearance of
characteristic, when two different influenza viruses infect a a new influenza strain before adequate amounts of vaccine can be
cell at the same time, the progeny produced can have RNA manufactured. However, these multivalent vaccines can be 80% to
segments from either of the viruses. From an infectious dis- 90% effective in preventing influenza. Because influenza vaccines
ease standpoint, this is particularly a problem when a genome are inactivated, they are relatively safe even for the immunocom-
segment from a virus that normally infects only a non-human promised. An attenuated vaccine that is given as a nasal spray to
host is acquired by a strain that infects humans. For example, children has been developed and is safe and effective. The main
if a pig is simultaneously infected with two virus strains—one features of influenza are summarized in table 21.11.
that normally infects birds and pigs and another that usually
infects only pigs and humans—the viruses that emerge will
still have eight genome segments, but the segments may Respiratory Syncytial Virus Infections
originate from either of the initial infecting strains. This can Respiratory syncytial virus (RSV) infection is the leading cause
result in a human-infecting strain that has novel HA and/or of serious lower respiratory tract infections in infants and young
NA antigens for which populations have no immunity (figure children, resulting in an estimated 90,000 hospitalizations and
21.24). In 2009, a new strain of H1N1 virus appeared that 4,500 deaths in the United States each year. It is also responsible
resulted from reassortment between bird, swine (pig), and for serious disease in elderly people, and for healthcare-associated
human viruses. This new strain spread quickly, causing a epidemics.
pandemic (swine flu). genetic reassortment, p. 320

Another situation arises when a strain that does not typically Signs and Symptoms
infect humans gains the ability to do so. Ecological studies show Signs and symptoms of RSV infection begin after an incubation
that all the known influenza A virus types exist in aquatic birds, period of 1 to 4 days with runny nose followed by cough, wheez-
generally causing chronic intestinal infections. Bird influenza ing, and difficulty breathing. Fever may or may not be present.
viruses (avian flu) readily infect domestic fowl, and from them can Patients often develop a dusky color, indicating that they are not
infect other domestic animals and humans. The 1997 “bird flu” epi- getting enough O2. Healthy older children and adults with RSV
demic in Hong Kong involved an H5N1 virus from chickens that generally show symptoms of a bad cold. RSV is one of the causes
spread to humans, sometimes causing fatal infections. Different of croup, which manifests as a loud high-pitched cough and noisy
H5N1 strains have been categorized as low pathogenic avian inhalation due to airway obstruction.
influenza (LPAI) and highly pathogenic avian influenza (HPAI), Hospitalized infants seldom die from RSV, but it is some-
depending on whether they cause fatal disease or not. Fortunately, times fatal for elderly patients with underlying diseases such as
the HPAI strain that started the Hong Kong outbreak did not spread heart and lung disease, cancer, and immunodeficiency.
easily from person to person and so the number of human cases
remained relatively low. Subsequent outbreaks show that HPAI Causative Agent
H5N1 viruses are now endemic in a number of Asian communities. RSV is a single-stranded, enveloped RNA virus of the paramyxo-
LPAI viruses have been isolated in the United States. A constant virus group. It causes cells in cell cultures to fuse together; the
concern is that an HPAI strain will evolve through antigenic drift clumps of fused cells are known as syncytia, thus the name of
or antigenic shift to spread person to person. the virus. paramyxovirus family, p. 309
512 Chapter 21 Respiratory System Infections

TABLE 21.11 Influenza

1 Influenza virus is inhaled and Signs and Fever, muscle aches, lack of energy,
carried to the lungs. 6 symptoms headache, sore throat, nasal
congestion, cough
2 Viral hemagglutinin attaches 1 6
to specific receptors on Incubation 1 to 2 days
ciliated epithelial cells, the 7 period
viral envelope fuses with the
epithelial cell, and the virus Causative agent Influenza virus, an orthomyxovirus
enters the cell by endocytosis. 1 Pathogenesis Infection of respiratory epithelium;
3 Host cell synthesis is diverted cells destroyed and virus released to
to synthesizing new virus. infect other cells. Secondary bacterial
6
infection results from damaged
4 Newly formed virions bud mucociliary escalator.
from infected cells; they are
released by viral neuraminidase Epidemiology Antigenic drift and antigenic shift
and infect ciliated epithelium, prevent immunity.
mucus-secreting, and alveolar
Treatment and Amantadine and rimantadine are
cells.
prevention sometimes effective for preventing
5 Infected cells ultimately die type A but not type B virus disease;
and slough off; recovery of the neuraminidase inhibitors effective
mucociliary escalator may take against both A and B viruses. These
weeks. medications somewhat effective for
treatment when given early in the
6 Secondary bacterial infection
disease.
of the lungs, ears, and sinuses
is common. Vaccines usually 80% to 90% effective.
7 The virus exits with coughing.

Pathogenesis Hantavirus Pulmonary Syndrome

The virus enters the body by inhalation and infects the respiratory In the spring of 1993, a newly emerging disease made a dra-
tract epithelium, causing cells to die and slough off. Bronchiolitis matic appearance in the “Four Corners” region of the American
is a common feature of the disease; the inflamed bronchioles Southwest, an area where the states of Arizona, Colorado, New
become partially plugged by sloughed cells, mucus, and clot- Mexico, and Utah come together. It was a small outbreak but quite
ted plasma that has oozed from the walls of the bronchi. The alarming because most victims were vigorous young adults who
initial obstruction causes wheezing when air rushes through the developed influenza-like symptoms, and then died within days.
narrowed passageways, sometimes causing the condition to be Scientists from the CDC rushed to join local epidemiologists and
confused with asthma. The obstruction often acts like a one-way
valve, allowing air to enter the lungs, but not leave them. In many
cases the inflammatory process extends into the alveoli, causing Respiratory Syncytial
pneumonia. There is a high risk of secondary infection because TABLE 21.12
Virus (RSV) Infections
of the damaged mucociliary escalator. mucociliary escalator, p. 337
Signs and Runny nose, cough, fever, wheezing,
Epidemiology symptoms difficulty breathing, dusky color

RSV outbreaks are common from late fall to late spring, peaking Incubation 1 to 4 days
period
in mid-winter. Recovery from infection produces only weak and
short-lived immunity, so that infections can recur throughout life. Causative agent RSV, a paramyxovirus that produces syncytia
Healthy children and adults usually have mild illness and readily Pathogenesis Sloughing of respiratory epithelium and
spread the virus to others. inflammatory response plug bronchioles,
cause bronchiolitis; pneumonia results from
bronchiolar and alveolar inflammation, or
Treatment and Prevention
secondary infection.
There are no effective antiviral medications for treating RSV.
Epidemiology Yearly epidemics during the cool months;
Preventing healthcare-associated RSV illness requires strict iso- readily spread by otherwise healthy older
lation techniques. People with underlying illnesses can be protected children and adults who often have mild
from the disease by giving them passive immunity via monthly symptoms; no lasting immunity.
injections of immune globulin or a monoclonal antibody called Treatment No satisfactory antiviral treatment. No
palivizumab. No vaccines are available, although there are at least six and prevention vaccine. Preventable by injections of immune
programs underway to develop a vaccine. The main features of RSV serum globulin or a monoclonal antibody.
infections are summarized in table 21.12. passive immunity, p. 420
 Part IV Infectious Diseases 513

health officials investigating the outbreak. Their studies quickly 50

established that the disease was associated with exposure to
mice. They soon learned that the patients had been infected with 45
a hantavirus closely related to one that plagued American troops
40
during the 1950s Korean War.
35
Signs and Symptoms

Number of cases
Hantavirus pulmonary syndrome usually begins with fever, 30
muscle aches (especially in the lower back), nausea, vomiting, and
25
diarrhea. Unproductive cough and increasingly severe shortness
of breath appear within a few days, followed by shock and death. 20

Causative Agents 15
Certain hantaviruses, enveloped viruses of the bunyavirus family,
10
cause hantavirus pulmonary syndrome. Their genome consists of
three segments of single-stranded RNA. In nature, these viruses 5
primarily infect rodents, causing lifetime infections, without any
apparent harm to the animals. Each type of hantavirus generally 0
infects a particular rodent species. bunyavirus family, p. 309 1998 2000 2002 2004 2006 2008 2009
Year
Pathogenesis
FIGURE 21.25 Total Hantavirus Pulmonary Syndrome Cases,
The virus enters the body by inhalation of airborne dust contami- United States, 1996–2009
nated with the urine, feces, or saliva of infected rodents. The virus
? Which animals most commonly carry hantaviruses?
then enters the circulation and is carried throughout the body,
infecting the cells that line tissue capillaries. Massive amounts of
the viral antigen appear in lung capillaries, but the antigen is also
Prevention of the syndrome is based on minimizing exposure
found in capillaries of the heart and other organs. The inflamma-
to rodents and dusts contaminated by their urine, saliva, and feces.
tory response to the viral antigen causes the capillaries to leak
Rodent populations should be controlled by keeping foods in con-
large amounts of plasma into the lungs, suffocating the patient
tainers and making buildings as mouse-proof as possible. When
and causing the blood pressure to fall. Shock and death occur in
cleaning an area where rodents are found, maximal ventilation
more than 40% of the cases. Luckily, despite the large amount of
should be ensured and the area should be mopped with a disin-
viral antigen in the lung capillaries, few mature infectious virions
fectant solution rather than being swept with brooms and vacuum
enter the air passages of the lung, so person-to-person transmis-
cleaners that can stir up dust. Lethal traps and poisons may be
sion occurs rarely, if ever.
necessary to decrease the rodent population in the area. The main
features of hantavirus pulmonary syndrome are summarized in
Epidemiology
table 21.13.
Hantavirus pulmonary syndrome is a zoonosis. It is considered
an emerging disease because of its recent discovery and apparent
increase in frequency but has probably existed for centuries. Since
the description of the syndrome, cases have been identified from TABLE 21.13 Hantavirus Pulmonary Syndrome
Canada to Argentina, including several hundred from the United
States (figure 21.25). Most of the cases have occurred west of the Signs and Fever, muscle aches, vomiting, diarrhea,
symptoms cough, shortness of breath, shock
Mississippi River and were due to a type of hantavirus carried
by deer mice, but hantavirus carried by other rodents can cause Incubation 3 days to 6 weeks
period
disease as well. Epidemics have been associated with increases in
mouse populations in poor communities with substandard hous- Causative agent Sin Nombre and related hantaviruses of
the bunyavirus family
ing. Thirty percent or more of the mice can become carriers of
the disease. Complex ecological factors such as numbers of foxes, Pathogenesis Viral antigen localizes in capillary walls in
owls, snakes, and other predators, and weather (which affect food the lungs; inflammation.
supply) all play a role in mouse population levels. The emergence Epidemiology Zoonosis likely to involve humans in
of hantavirus pulmonary syndrome is a convincing example of proximity to increasing mouse populations;
generally no person-to-person spread.
how environmental change can result in infectious human disease.
zoonoses, p. 439 emerging diseases, p. 448 Treatment Avoid contact with rodents; seal access to
and prevention houses, food supplies; good ventilation,
avoid dust, use disinfectants in cleaning
Treatment and Prevention rodent-contaminated areas. No proven
There is no proven antiviral treatment for hantavirus pulmonary antiviral treatment.
syndrome, a highly fatal disease.
514 Chapter 21 Respiratory System Infections

MicroAssessment 21.5
The speed with which influenza travels around the world and the
potential for development of virulent influenza virus strains make the
disease an extremely serious threat to humankind. Most deaths from
influenza are caused by secondary bacterial infections. Respiratory
syncytial virus is the leading cause of serious respiratory disease in
infants and young children. Hantavirus pulmonary syndrome, first
recognized in 1993, is often fatal. It is contracted from inhalation of
dust contaminated by urine, feces, or saliva from mice infected with
certain hantaviruses.
13. Why are there so many deaths from influenza when it is
generally a mild disease?
14. What is the source of the virus that causes hantavirus
pulmonary syndrome? (a) 20 μm
15. Why might you expect an influenza epidemic to be more
severe following an antigenic shift in the virus than after
antigenic drift? +

21.6 ■ Fungal Infections

of the Lung
Learning Outcomes
11. Describe the pathogenesis of histoplasmosis.
12. Outline the epidemiology of coccidioidomycosis

Serious lung diseases caused by fungi are quite unusual in healthy,

immunocompetent individuals. Symptomatic and asymptomatic (b) 50 μm
infections that subside without treatment, however, are common. FIGURE 21.26 Coccidioides immitis (a) Mold-phase hyphae
Coccidioidomycosis and histoplasmosis are two examples of fragmenting into arthroconidia. (b) Spherules containing fungal
widespread mycoses of the respiratory tract. endospores.

MicroByte ? Which form of this fungus is found in host tissues?

Coccidioidomycosis and histoplasmosis can both manifest with
symptoms that resemble tuberculosis.
thick-walled spherules that may contain several hundred small
cells called endospores, not to be confused with bacterial endo-
Coccidioidomycosis (“Valley Fever”) spores (figure 21.26b). dimorphic fungi, p. 285
In the United States, coccidioidomycosis occurs mainly in
California, Arizona, Nevada, New Mexico, Utah, and West Texas. Pathogenesis
People who are exposed to dust and soil, such as farm workers, are Arthroconidia enter the lung with inhaled air and develop into
most likely to become infected, but only 40% develop symptoms. spherules. Endospores develop in the spherules, which mature
and rupture, releasing them. The endospores develop into more
Signs and Symptoms endospore-containing spherules and the process is repeated. Each
“Flulike” signs and symptoms such as fever, cough, chest pain, time this cycle occurs, an inflammatory response is provoked.
and loss of appetite and weight are common manifestations of coc- Tissue injury and symptoms are caused mainly by the host’s
cidioidomycosis. The majority of people with the disease recover immune response to coccidioidal antigens.
spontaneously within a month. A small percentage of patients, The organisms are usually eliminated by body defenses, but
however, develop chronic disease. caseous necrosis (dead tissue that has cheeselike consistency)
occurs in a small percentage of individuals, resulting in lung cavi-
Causative Agent ties similar to those seen in tuberculosis. Occasionally, organisms
Coccidioidomycosis is caused by Coccidioides immitis, a dimor- are carried throughout the body by the bloodstream and infect
phic fungus that grows in soil as a mold. Its hyphae give rise the skin, mucous membranes, brain, and other organs. This dis-
to numerous barrel-shaped, highly infectious structures called seminated form of the disease occurs more often in people with
arthroconidia (figure  21.26a). These become airborne and can AIDS or other immunodeficiencies and is fatal without treatment.
be inhaled. In infected tissues, the arthroconidia develop into caseous necrosis, p. 504
 Part IV Infectious Diseases 515

Epidemiology Signs and Symptoms

Coccidioides immitis grows only in semi-arid desert areas of the Most infections are asymptomatic. Fever, cough, and chest pain
Western Hemisphere. In these areas, infections occur only during are the most common symptoms, sometimes with shortness of
the hot, dry, dusty seasons when airborne arthroconidia are easily breath. Mouth sores may develop, especially in children.
dispersed from the soil. Dust stirred up by earthquakes can result
in epidemics. Rainfall encourages growth of the fungus, which Causative Agent
then produces increased numbers of spores when dry conditions Histoplasmosis is caused by the dimorphic fungus Histoplasma
return. People can contract coccidioidomycosis by simply travel- capsulatum, although the name is misleading because the fun-
ing through the endemic area. Infectious spores have unknowingly gus does not have a capsule. This organism grows in soils con-
been transported to other areas, but the organism apparently is taminated by bat or bird droppings, but it is not pathogenic for
unable to establish itself in moist climates. these animals. In pus or tissue from people with active disease,
H. capsulatum is a tiny oval yeast that grows within host macro-
Treatment and Prevention phages (figure 21.27a). The mold form of the organism charac-
Medications approved for treatment of serious cases of coc- teristically produces two kinds of conidia: macroconidia, which
cidioidomycosis include amphotericin B and fluconazole or often have numerous projecting knobs (figure  21.27b), and tiny
itraconazole. They must be given for long periods of time, and pear-shaped or spherical microconidia.
cause troublesome side effects. Even with treatment, disseminated
disease can reactivate months or years later.
Preventive measures include avoiding dust in the endemic Macrophage
areas, and watering and planting vegetation to aid in dust control. nucleus Macrophage
Table 21.14 describes the main features of coccidioidomycosis.

Histoplasmosis (“Spelunker’s Disease”)

Histoplasmosis, like coccidioidomycosis, is usually benign but
occasionally mimics tuberculosis. Rare, serious forms of the dis-
ease suggest that the patient has an underlying immunodeficiency
such as AIDS. The distribution is more widespread than that of
coccidioidomycosis and is associated with different soil types
and climate.
H. capsulatum

TABLE 21.14 Coccidioidomycosis (Valley Fever)

Signs and Fever, cough, chest pain, loss of appetite
(a) 10 μm
symptoms and weight; less frequently, painful
nodules on extremities, pain in joints; skin,
mucous membranes, brain, and internal
organs sometimes involved
Incubation period 2 days to 3 weeks

Causative agent Coccidioides immitis, a dimorphic fungus

Pathogenesis After lodging in lung, arthrospores develop
into spherules that mature and discharge
endospores, each of which then develops
into another spherule; inflammatory
response damages tissue; hypersensitivity
to fungal antigens causes painful nodules
and joint pain.
Epidemiology Inhalation of airborne C. immitis spores
with dust from soil growing the organism.
Occurs only in certain semi-arid regions of
the Western Hemisphere.
(b) 10 μm
Treatment Treatment: amphotericin B and
and prevention fluconazole or itraconazole. Prevention
FIGURE 21.27 Histoplasma capsulatum (a) Yeast-phase
organisms in the cytoplasm of a macrophage. (b) Mold phase,
by dust control methods such as grass
showing macroconidia.
planting and watering.
? Is this fungus encapsulated?
516 Chapter 21 Respiratory System Infections

Pathogenesis Histoplasmosis
Histoplasma capsulatum conidia inhaled into the lungs are taken TABLE 21.15
(Spelunker’s Disease)
up by resident macrophages. The fungus then develops into the
yeast form, which multiplies within the phagocytes. Granulomas Signs and Mild respiratory symptoms; less frequently,
symptoms fever, chest pain, cough, chronic sores
develop in infected areas, closely resembling those seen in tuber-
culosis, sometimes even showing caseous necrosis. Eventually, Incubation period 5 to 8 days
the lesions are replaced with scar tissue, and many calcify, becom- Causative agent Histoplasma capsulatum, a dimorphic fungus
ing visible on X rays. In rare cases, particularly in those who are Pathogenesis Spores inhaled, change to yeast phase,
immunodeficient, the disease spreads throughout the body. multiply in macrophages; granulomas form;
disease spreads in individuals with AIDS or
Epidemiology other immunodeficiencies.

The distribution of histoplasmosis is quite different from that Epidemiology The fungus prefers to grow in soil
of coccidioidomycosis. Figure  21.28 shows the distribution of contaminated by bird or bat droppings,
especially in Ohio and Mississippi River valleys,
histoplasmosis in the United States, but the disease also occurs and in the U.S. Southeast. Spotty distribution
in tropical and temperate zones scattered around the world. Cave in many other countries around the world.
explorers (spelunkers) are at risk for histoplasmosis because Spelunkers are at risk of infection.
many caves contain soil contaminated with bat droppings. Most Treatment Treatment: amphotericin B and itraconazole
cases in the United States have occurred in the Mississippi and and prevention for serious infections. Prevented by
Ohio River drainage area and in South Atlantic states. Skin tests avoiding soils contaminated with chicken,
reveal that millions of people living in these areas have been bird, or bat droppings.
infected.

Treatment and Prevention

Treatment of histoplasmosis is similar to that of coccidioido- No proven preventive measures are known other than to
mycosis. Amphotericin B and itraconazole are used for treating avoid areas where soil is heavily enriched with bat and bird
severe disease, but both medications have potentially serious droppings, especially if the soil has been left undisturbed for a
side effects. long period. Some researchers have recommended placing sev-
eral inches of clay soil over soils containing large quantities of
old droppings. The main features of histoplasmosis are described
in table 21.15.

The key features of the diseases covered in this chapter are high-
lighted in the Diseases in Review 21.1 table.

MicroAssessment 21.6
Coccidioidomycosis and histoplasmosis are two diseases caused by
fungi that live in the soil. The body responds to these infections by
forming granulomas, mimicking tuberculosis. Each fungus has its
own ecological niche: Coccidioides immitis in semi-arid regions of
the Western Hemisphere, and Histoplasma capsulatum in moist soils
enriched with bird or bat droppings around the world.
16. Why should an immunodeficient person avoid traveling through
hot, dry, dusty areas of the Southwest?
17. Why might cave exploration increase the risk of histoplasmosis?
FIGURE 21.28 Geographical Distribution of Histoplasma 18. Several students staying in a hotel next to a bulldozing operation
capsulatum in the United States developed histoplasmosis. How might the bulldozing explain the
outbreak? +
? Why is histoplasmosis also called “spelunker’s disease”?
Diseases in Review 21.1
Diseases of the Respiratory System

Disease Causative Agent Comment Summary Table

BACTERIAL INFECTIONS OF THE UPPER RESPIRATORY TRACT

Streptococcal pharyngitis Streptococcus pyogenes Treated with antibiotics, partly to avoid sequelae; must be Table 21.3, p. 489
(“strep throat”) (group A streptococcus) distinguished from viral pharyngitis, which cannot be treated
with antibiotics.
Diphtheria Corynebacterium Toxin-mediated disease characterized by pseudomembrane in Table 21.4, p. 492
diphtheriae the upper respiratory tract. Preventable by vaccination (DTaP).
Conjunctivitis (pinkeye), Usually Haemophilus Often occur together; factors involved in the transmission are
otitis media (earache), influenzae and unknown.
sinus infection Streptococcus pneumoniae

VIRAL INFECTIONS OF THE UPPER RESPIRATORY TRACT

Common cold Rhinoviruses and other Runny nose, sore throat, and cough are due to the Table 21.5, p. 495
viruses inflammatory response and cell destruction.
Adenoviral pharyngitis Adenovirus Similar to the common cold but with fever; spread to the lower Table 21.6, p. 496
respiratory tract can result in severe disease.

BACTERIAL INFECTIONS OF THE LOWER RESPIRATORY TRACT

Pneumococcal pneumonia Streptococcus Organism common in the throat of healthy people; causes Table 21.7, p. 499
pneumoniae disease when mucociliary escalator is impaired or with
underlying conditions. Vaccine that protects against multiple
strains is available.
Klebsiella pneumonia Klebsiella species, commonly Common hospital-acquired bacterium; characterized by sputum Table 21.7, p. 499
K. pneumoniae resembling red current jelly. Drug resistance is a major problem.
Mycoplasmal pneumonia Mycoplasma pneumoniae Relatively mild pneumonia; common among college students Table 21.7, p. 499
(“walking pneumonia”) and military recruits. Cannot be treated with medications that
inhibit cell wall synthesis.
Pertussis Bordetella pertussis Characterized by frequent violent coughing. Preventable by Table 21.8, p. 502
(“whooping cough”) vaccination (DTaP).
Tuberculosis (“TB”) Mycobacterium Most infections result in latent tuberculosis infection (LTBI), but Table 21.9, p. 506
tuberculosis these can reactivate to cause active tuberculosis disease (ATBD).
Treated using combination drug therapy, but drug resistance is
an increasing problem.
Legionnaires’ disease Legionella Transmitted via aerosolized water drops; smokers and those Table 21.10, p. 507
pneumophila with impaired defenses are most at risk of developing disease.

VIRAL INFECTIONS OF THE LOWER RESPIRATORY TRACT

Influenza (“flu”) Influenza A virus New vaccine developed yearly; viruses change seasonally due Table 21.11, p. 512
to antigenic drift; antigenic shifts cause pandemics.
Respiratory syncytial RSV Serious disease in infants, young children, and the elderly. Table 21.12, p. 512
virus infections
Hantavirus pulmonary Hantaviruses Acquired via inhaled dust contaminated with rodent saliva, Table 21.13, p. 513
syndrome urine, or feces. Frequently fatal.

FUNGAL INFECTIONS OF THE RESPIRATORY TRACT

Coccidioidomycosis Coccidioides immitis Environmental reservoir (soil in semi-arid desert areas); most Table 21.14, p. 515
(“Valley fever”) infections are asymptomatic.
Histoplasmosis Histoplasma capsulatum Environmental reservoir (soil enriched with bird or bat Table 21.15, p. 516
(“spelunker’s disease”) droppings); most infections are asymptomatic.
518 Chapter 21 Respiratory System Infections

FUTURE CHALLENGES 21.1

Global Preparedness Versus Emerging Respiratory Viruses
In November 2002, a frightening new respi- proved to be a previously unknown corona- waterfowl that readily infect domestic flocks,
ratory disease emerged in Guangdong prov- virus, and its source was never identified, which then infect humans. Epidemic influenza
ince, China. Symptoms included cough and although it was probably a wild animal sold for in humans generally has a short incubation
fever, and X rays showed characteristics of meat in the markets. On three occasions, SARS period, and many victims have mild, coldlike
viral pneumonia. The disease, labeled “SARS” escaped from laboratories where the virus was symptoms not easily recognized as part of an
(severe acute respiratory syndrome), quickly being studied, and the disease may arise again influenza epidemic. Therefore, quarantines are
spread around the world, involving 25 coun- from its natural source. However, its main not helpful in disease control. Many experts
tries and causing more than 700 deaths. Many importance is that it caused many countries think we have been lucky that no virus capable
of its victims were medical personnel. It had to learn to work together to help forge a more of causing a pandemic has appeared. However,
two characteristics that quickly led to its con- effective global response to emerging pan- thanks to the battles against the H5N1 avian
trol: a long incubation of 6 days, and dramatic demic diseases. More details on SARS can be virus and the SARS outbreaks, the world is
symptoms that were easily recognized. This found at www.sarsreference.com. much closer to being prepared for the next pan-
allowed healthcare personnel time to identify The problem of avian influenza is more demic. The challenge is to sustain and improve
contacts and institute quarantines to stop the difficult to solve than SARS because the avian advances in cooperative surveillance, and
spread of the disease. The causative agent viruses are carried in the intestines of wild action planning to avoid a global catastrophe.

Summary
21.1 ■ Anatomy, Physiology, and Ecology a milder illness. Otitis media and sinusitis develop when infection
spreads from the nasopharynx (figure 21.9).
The Upper Respiratory Tract
The upper respiratory tract includes the nose and nasal cavity, phar- 21.3 ■ Viral Infections of the Upper Respiratory System
ynx (throat), and epiglottis (figure 21.1). Ciliated cells line much of the
The Common Cold (table 21.5)
respiratory tract and remove microorganisms by constantly propelling
mucus out of the respiratory system. A wide variety of microorganisms The common cold can be caused by many different viruses, rhinoviruses
colonize parts of the system (table 21.1). being the most common (figure 21.10).

The Lower Respiratory Tract Adenoviral Respiratory Tract Infections (table 21.6)

The lower respiratory system includes the larynx, trachea, bronchi, and Adenoviruses cause illnesses that can resemble a common cold or strep
lungs. Pleural membranes surround the lungs. Viruses and microorgan- throat, with symptoms varying from mild to severe.
isms are normally absent from the lower respiratory system.
INFECTIONS OF THE LOWER
INFECTIONS OF THE UPPER RESPIRATORY SYSTEM
RESPIRATORY SYSTEM
21.4 ■ Bacterial Infections of the Lower
21.2 ■ Bacterial Infections of the Upper Respiratory System
Respiratory System
Pneumococcal Pneumonia (table 21.7; figures 21.11, 21.12)
Streptococcal Pharyngitis (“Strep Throat”) (tables 21.2, 21.3) Streptococcus pneumoniae, one of the most common causes of pneumo-
Streptococcus pyogenes is a β-hemolytic Gram-positive coccus that nia, is virulent because of its capsule.
causes strep throat (figures 21.2, 21.3). Strains that produce SPEs
Klebsiella Pneumonia (table 21.7)
(streptococcal pyrogenic exotoxins) can cause scarlet fever. Other
diseases are associated with SPEs as well. Klebsiella pneumonia is representative of many healthcare-associated
pneumonias that cause permanent damage to the lung (figure  21.13).
Post-Streptococcal Sequelae Serious complications such as lung abscesses and bloodstream infection
Post-streptococcal sequelae, including rheumatic fever and glomerulo- are more common than with many other bacterial pneumonias.
nephritis, may follow strep throat and are due to the immune response
Mycoplasmal Pneumonia (“Walking Pneumonia”)
and molecular mimicry (figure 21.5).
(table 21.7, figure 21.14)
Diphtheria (table 21.4) Mycoplasmal pneumonia is often called walking pneumonia; serious
Diphtheria, caused by Corynebacterium diphtheriae, is a toxin-mediated complications are rare. Penicillins and cephalosporins are not useful in
disease that can be prevented by immunization (figures 21.6, 21.7). treatment because the causative agent, M. pneumoniae, lacks a cell wall.
Pinkeye, Earache, and Sinus Infections Pertussis (“Whooping Cough”) (table 21.8; figures 21.15, 21.16)
Conjunctivitis (pinkeye) is usually caused by Haemophilus influenzae Whooping cough is characterized by violent spasms of coughing and
or Streptococcus pneumoniae (pneumococcus) (figure  21.8). Viral gasping. Childhood immunization against the causative agent, Bordetella
agents, including adenoviruses and rhinoviruses, usually result in pertussis, prevents the disease. Mild disease is common in adults.
 Part IV Infectious Diseases 519

Tuberculosis (“TB”) (table 21.9; figures 21.17–21.21) Hantavirus Pulmonary Syndrome (table 21.13, figure 21.25)
Tuberculosis, caused by the acid-fast rod Mycobacterium tuberculosis, Hantavirus pulmonary syndrome is an often fatal disease contracted
is generally slowly progressive or heals and remains latent, presenting when airborne dust contaminated by urine from mice infected with a
the risk of later reactivation. hantavirus is inhaled.
Legionnaires’ Disease (table 21.10, figure 21.22) 21.6 ■ Fungal Infections of the Lung
Legionnaires’ disease occurs when there is a high infecting dose of the
causative microorganisms or an underlying lung disease. The cause, Coccidioidomycosis (“Valley Fever”) (table 21.14)
Legionella pneumophila, is a rod-shaped bacterium common in the Coccidioidomycosis occurs in hot, dry areas of the Western Hemisphere
environment. and is initiated by airborne spores of the dimorphic soil fungus
Coccidioides immitis (figure 21.26).
21.5 ■ Viral Infections of the Lower Respiratory System
Histoplasmosis (“Spelunker’s Disease”) (table 21.15)
Influenza (“Flu”) (table 21.11; figures 21.23, 21.24) Histoplasmosis is similar to coccidioidomycosis but occurs in tropi-
Widespread epidemics are characteristic of influenza A viruses. cal and temperate zones around the world (figure 21.28). The causative
Antigenic shifts and drifts are responsible. Deaths are usually but not fungus, Histoplasma capsulatum, is dimorphic and is found in soils
always caused by secondary infection. Reye’s syndrome may rarely contaminated by bat or bird droppings (figure 21.27).
occur during recovery from influenza and other viral infections but is
probably not caused by the virus itself.
Respiratory Syncytial Virus Infections (table 21.12)
RSV is the leading cause of serious respiratory disease in infants and
young children.

Review Questions
Short Answer 3. Adenoviral infections generally differ from the common cold in all
1. How does contamination of the eye lead to upper respiratory the following ways, except adenoviral infections are
infection? a) not caused by picornaviruses.
2. After you recover from strep throat, can you get it again? Explain. b) often associated with fever.
3. Where is the gene for diphtheria toxin production located? c) associated with severe sore throat.
4. Describe two ways to decrease the chance of contracting a cold. d) much more likely to cause pneumonia.
e) avoided by handwashing.
5. What kinds of diseases are caused by adenoviruses?
4. All are true of mycoplasmal pneumonia except
6. How do alcoholism and cigarette smoking predispose a person to
pneumonia? a) it is a mycosis.
7. Give a mechanism by which Klebsiella sp. become antibiotic- b) it usually does not require hospitalization.
resistant. c) penicillin is ineffective for treatment.
8. Why does the incidence of whooping cough rise promptly when d) it is the leading cause of bacterial pneumonia in college
pertussis immunizations are stopped? students.
e) the infectious dose of the causative organism is low.
9. Why are two or more antitubercular medications used together to
treat tuberculosis? 5. All of the following are true of Legionnaires’ disease except
10. Why did it take so long to discover the cause of Legionnaires’ a) the causative organism can grow inside amebas.
disease? b) it spreads readily from person to person.
c) it is more likely to occur in long-term cigarette smokers than in
Multiple Choice nonsmokers.
1. The following are all complications of streptococcal pharyngitis d) it is often associated with diarrhea or other intestinal symptoms.
except e) it can be contracted from household water supplies.
a) glomerulonephritis. 6. Which of the following infectious agents is most likely to cause a
b) scarlet fever. pandemic?
c) subacute bacterial endocarditis. a) Influenza A virus
d) acute rheumatic fever. b) Streptococcus pyogenes
e) Reye’s syndrome. c) Histoplasma capsulatum
2. All of the following are true of diphtheria except d) Sin Nombre virus
a) a membrane that forms in the throat can cause suffocation. e) Coccidioides immitis
b) a toxin is produced that interferes with ribosome function.
c) the causative organism typically invades the bloodstream.
d) immunization with a toxoid prevents the disease.
e) nerve injury with paralysis is common.
520 Chapter 21 Respiratory System Infections

7. Respiratory syncytial virus Applications

a) is a leading cause of bronchiolitis in infants. 1. A physician is advising the family on the condition of a diphtheria
b) is an enveloped DNA virus of the adenovirus family. patient. How would the physician explain why the disease affects
c) attaches to host cell membranes by means of neuraminidase. some tissues and not others?
d) poses no threat to elderly people. 2. How should a physician respond to a mother who asks if her
e) mainly causes disease in the summer months. daughter can get pneumococcal pneumonia again?
8. In the United States, hantaviruses
a) are limited to southwestern states.
Critical Thinking +
b) are carried only by deer mice. 1. If all transmission of Mycobacterium tuberculosis from one person
c) infect human beings with a fatality rate above 40%. to another was stopped, how long would it take for the world to be
rid of the disease?
d) were first identified in the early 1970s.
e) are contracted mainly in bat caves. 2. Medications that prevent and treat influenza by binding to neur-
aminidase on the viral surface act against all the kinds of influenza
9. All of the following are true of coccidioidomycosis except
viruses that infect humans. What does this imply about the nature
a) it is contracted by inhaling arthrospores. of the interaction between the medications and the neuraminidase
b) it is caused by a dimorphic fungus. molecules?
c) endospores are produced within a spherule.
d) it is more common in Maryland than in California.
e) it is often associated with painful nodules on the legs.
10. The disease histoplasmosis
a) is caused by an encapsulated bacterium.
b) is contracted by inhaling arthrospores.
c) occurs mostly in hot, dry, and dusty areas of the American
Southwest.
d) is a threat to AIDS patients living in areas bordering the
Mississippi River.
e) is commonly fatal for pigeons and bats.
22 Skin Infections
KKEEYYTTEERRMMS S
Abscess A localized collection of
pus within a tissue.
Carbuncle Painful infection of
Furuncle A boil; a localized
skin infection that penetrates into
subcutaneous tissue.
the skin and subcutaneous tissues; Impetigo A superficial skin
manifests as a cluster of boils. infection characterized by thin-
walled vesicles, oozing blisters,
Exfoliatin A bacterial toxin
and yellow crusts; caused by
that causes sloughing of the outer
Staphylococcus aureus and
epidermis.
Streptococcus pyogenes.
Folliculitis Inflammation of a hair
Pyoderma Any skin disease
follicle.
characterized by production of pus.

A dividing Staphylococcus epidermidis cell. scientific name and went off to Mexico to study a very similar disease—
louse-borne typhus. Unfortunately, he contracted typhus and died at the
age of 39. Stanislaus Prowazek, a European scientist studying the same
disease, met the same fate at almost the same age.
The martyrdom of the two young scientists struggling to understand
A Glimpse of History infectious diseases is memorialized in the name of the louse-borne typhus
Howard T. Ricketts was born in Ohio in 1871. He studied medicine in agent, Rickettsia prowazekii. Both the genus and species names of the
Chicago, and then specialized in pathology, the study of the nature of RMSF agent, Rickettsia rickettsii, recognize Howard Ricketts. We now
disease and its causes. In 1902, he was appointed to the faculty of the know that these bacteria are obligate intracellular parasites. Since most
University of Chicago, where his research interests turned to Rocky culture media lack live cells, the bacteria did not grow in the culture
Mountain spotted fever (RMSF). media used by Ricketts and Prowazek.
While patients with RMSF have a dramatic rash and often die, the
disease was poorly understood. Ricketts noticed that people and labora-

M
tory animals with it had tiny rod-shaped bacteria in their blood. He was uch of the human body’s contact with the outside world
sure these tiny invaders caused the disease, because he could transmit occurs at the skin surface. This tough, flexible outer
the disease by injecting laboratory animals with blood from an infected covering provides a remarkable barrier to invasion and
person. However, he was never able to grow the bacteria on laboratory infection. Its exposed state, however, leaves it vulnerable to a vari-
media. ety of injuries. Cuts, punctures, burns, chemical injury, and insect
Based on observations of patients with RMSF, scientists suspected or tick bites can break this barrier and provide a way for pathogens
that the disease was contracted from tick bites. Ricketts went on to to enter underlying tissues. For example, Staphylococcus aureus
prove that certain species of ticks could transmit the disease from one
can enter a surgical wound and then invade the bloodstream,
animal to another. The infected ticks remained healthy but were capable
of transmitting the disease for long periods of time. Surprisingly, the
and sandfly bites can introduce Leishmania species, the cause of
offspring of infected ticks were often born infected as well. Ricketts leishmaniasis—more can be learned about this disease at the text
showed that the eggs of infected ticks frequently contained large num- website (www.mhhe.com/nester7). Skin infections also occur
bers of the same bacteria found in the patients’ blood. When these eggs when microorganisms or viruses that enter the body from another
were fertilized they developed into infected ticks. Ricketts was unable to site (such as the respiratory or gastrointestinal systems) are carried
cultivate these bacteria for further studies so he declined to give them a by the bloodstream to the skin.

521
522 Chapter 22 Skin Infections

22.1 ■ Anatomy, Physiology, contains many tiny nerves, glands, blood vessels, and lym-
phatic vessels. The dermis has connective tissue that binds
and Ecology irregularly to the fat and other cells that make up the subcuta-
neous layer.
Learning Outcomes
■ Subcutaneous tissue. While not part of the skin itself, the
1. Describe the function of skin in health and disease.
subcutaneous layer is made up of fat and other cells that sup-
2. Explain the role of normal skin microbiota in health port the layers of the skin.
and disease.
The outermost layers of skin are typically bathed in secretions.
The skin is far more than an inert wrapping for the body. It Glands in the dermis produce sweat and an oily secretion called
prevents the entry of microbes, regulates body temperature, and sebum. Sweat is delivered via fine tubules from the sweat glands
restricts the loss of fluid from body tissues. The multiple sensory to the surface of the skin where it evaporates to leave a salty resi-
receptors found in the skin provide the central nervous system due that inhibits many microbes. The sebaceous glands open into
with information about the environment. The skin also plays an hair follicles, so that sebum flows up through the follicles and out
essential role in the function of the immune system. If the bar- over the skin surface. Sebum keeps the hair and skin soft, pliable,
rier of the skin is breached, resident macrophages and dendritic and water-repellent.
cells produce cytokines that aid the development of an immune In addition to being a physical barrier, skin represents a
response. Collections of lymphocytes found in skin-associated distinct ecological habitat. Compared to the moist warm condi-
lymphoid tissues (SALT) are ready to proliferate when needed. tions in the respiratory tract, however, the skin is a cool desert.
cytokines, p. 341 skin-associated lymphoid tissue, p. 358 Members of the normal skin microbiota are uniquely suited to
The skin is composed of two layers—the epidermis and the thrive in this environment. They use the substances in sebum and
dermis (figure  22.1). The subcutaneous layer lies beneath the sweat as nutrients to fuel their growth, and in doing so, inhibit
dermis and supports the skin. other microbes. For example, resident bacteria degrade lipids in
sebum, producing fatty acids that are toxic to many bacteria. In
■ Epidermis. This is the surface layer, made up of multiple
fact, the skin surface is an unfriendly habitat for most pathogens,
layers of flat cells. The outermost cells are dead and filled
being too dry, salty, acidic, and toxic for their survival. Those
with keratin, a tough, water-resistant protein also found in
that tolerate the conditions are often shed with the dead skin cells.
hair and nails. These skin cells regularly flake off, exposing
normal microbiota, pp. 8, 381
their replacements immediately below. Cells at the boundary
between the dermis and epidermis are actively dividing. These
newly formed cells migrate to the skin’s surface, become flat-
tened and die as keratin fills their cytoplasm. Microbes living
on the outermost cells are shed with the flaking skin cells.
This process is a valuable defense against infection and
results in the complete regeneration of the epidermis
about once a month. Dandruff represents excessive
Hairs
shedding of skin cells.
■ Dermis. The dermis is the layer of skin cells
directly below the epidermis. This layer
Epidermis

Sebaceous Skin
gland

Arrector pili Dermis

(smooth muscle)

FIGURE 22.1 Microscopic Hair follicle

Anatomy of the Skin Notice Nerve
that the sebaceous unit, com-
Vein
posed of the hair follicle and
the attached sebaceous gland,
Artery Subcutaneous
almost reaches the subcutane-
Sweat gland tissue (hypodermis)
ous tissue.
Fat
? How can anaerobic bacteria,
such as Propionibacteria
species, grow on human
skin?
 Part IV Infectious Diseases 523

Although members of the normal microbiota play an essen- MicroAssessment 22.1

tial protective role on the skin, they can also be troublesome. For
The skin resists colonization by most microbial pathogens, provides
one thing, their metabolic products are responsible for body odor.
a physical barrier to infection, transmits sensory input from the
Sweat is odorless when first secreted, but bacteria degrade some environment, and assists the body’s regulation of temperature and
of its components, producing foul-smelling compounds. Most fluid balance. Members of the normal skin microbiota help protect
antiperspirants prevent body odor by decreasing the number and the skin from colonization by pathogens. Resident microbes are
metabolic activity of bacteria at the site of application. Some responsible for body odor and can cause disease when the host’s
members of the normal skin microbiota are opportunistic patho- defenses are impaired.
gens, causing disease in people with impaired body defenses. 1. Describe four characteristics of skin that help it resist infection.
opportunistic pathogens, p. 382 2. Name and describe three groups of microorganisms generally
Different regions of the skin can be compared to unique present on normal skin.
neighborhoods, made up of distinct numbers and types of inhab- 3. Would you expect a person living in the tropics or in the desert
itants. Depending on the body location and amount of moisture, to have larger numbers of bacteria living on the surface
the number of bacteria on the skin surface may range from of his or her skin? +
only about 1,000 organisms per square centimeter on the back
to more than 10 million in the groin and armpit, where there is
plenty of moisture. Most of the microbial skin inhabitants can
be categorized in three groups: diphtheroids, staphylococci, and
fungi (table 22.1). 22.2 ■ Bacterial Skin Diseases
In oily regions like the forehead, upper chest, and back,
diphtheroids are common. The name of this informal grouping Learning Outcomes
refers to the fact that these Gram-positive bacteria physically 3. Compare and contrast acne and hair follicle infections.
resemble Corynebacterium diphtheriae, the pathogen that causes 4. Describe the characteristics of staphylococcal scalded skin
diphtheria. Among the most common diphtheroids on the skin syndrome and impetigo.
are Propionibacterium species. These obligate anaerobes grow 5. Compare and contrast Lyme disease and Rocky Mountain
within hair follicles, where O2 is limited. Corynebacterium, p. 265 spotted fever.
diphtheroids, p. 265 anaerobe, p. 90
Staphylococci such as Staphylococcus epidermidis are com- Few bacterial species invade intact skin directly. However, strands
mon on the skin surface. These salt-tolerant Gram-positive cocci of hair provide a route of invasion by extending past the epidermis
grow well on the dry environment of the skin surface. Staphylo- to hair follicles that penetrate into the dermis. Acne and hair fol-
cocci use available nutrients on the skin, preventing pathogen licle infections both begin with the hair shaft and are two of the
colonization. They also produce antimicrobial substances highly most common infections due to direct invasion.
active against other Gram-positive bacteria, helping to maintain a
balance among the microbial inhabitants of the skin ecosystem.
Staphylococcus, p. 273
Acne Vulgaris
Malassezia species are tiny lipid-dependent yeasts (fungi) Acne in its most common form begins at puberty in association
present on human skin from late childhood onward. These yeasts with a rise in sex hormones. Although the incidence of acne drops
can be grown on laboratory media containing fatty substances dramatically after puberty, some individuals have acne well into
such as olive oil. their 30s and 40s.

MicroByte Signs and Symptoms

The average person sheds about 40,000 skin cells daily, or 1,500 Acne is characterized by enlarged sebaceous glands and increased
during a typical microbiology lecture. secretion of sebum. The hair follicle epithelium thickens and
sloughs off in clumps, gradually blocking the flow of sebum to the
skin surface. Continued sebum production by the infected gland
can force a plug of material to the surface, where it is visible as
TABLE 22.1 Principal Members of the Normal
a blackhead. With complete obstruction, the follicle fills with
Skin Microbiota
sebum, causing the epidermis to bulge outward. This produces a
Name Characteristics whitish lesion called a whitehead.
Diphtheroids Variably shaped, non-motile, Gram-positive
Causative Agent
rods of the Corynebacterium and
Propionibacterium genera The bacterium typically associated with acne is Propionibacterium
Staphylococci Gram-positive cocci arranged in packets or acnes. The cells are Gram-positive rods that grow anaerobically
clusters; coagulase negative; facultatively in and around hair follicles. They have lipases that break down
anaerobic the oily sebum in the sebaceous glands and use the resulting fatty
Fungi Small yeasts of the genus Malassezia that acids and glycerol as a food source. Sebum accumulates when a
require oily substances for growth gland is blocked, providing more food for P. acnes. These bacteria
then multiply to enormous numbers in the trapped sebum.
524 Chapter 22 Skin Infections

Pathogenesis Some Diseases Often Caused

The metabolic products of the dividing bacteria cause an inflam- TABLE 22.2
by Staphylococcus aureus
matory response, attracting neutrophils whose enzymes damage
Page for More
the wall of the enlarged follicle. This can cause the follicle to
Disease Information
burst, releasing its contents into the surrounding tissue. The result
is an abscess—a collection of pus surrounded by inflamed tissue. Carbuncles p. 524
The pus is made up of living and dead neutrophils, bacteria, and Endocarditis p. 672
tissue debris. Most abscesses will eventually heal, but may leave Folliculitis p. 524
a scar. inflammatory response, p. 348 neutrophils, p. 339
Food poisoning p. 757
Furuncles p. 524
Epidemiology
Impetigo p. 528
Most people have P. acnes on their skin throughout their lives.
The increased incidence of acne during puberty is probably due Scalded skin syndrome p. 527
to excess sebum production caused by increased hormone levels, Toxic shock syndrome p. 619
particularly testosterone. Infants may also have acne caused by Wound infections p. 551
maternal hormones stimulating sebum production.

Treatment and Prevention

Acne is usually a relatively mild infection that will run its course.
The length of infection can be limited by medications such to the arrangement of the bacteria as seen in stained smears. The
as antibiotics and benzoyl peroxide that inhibit the growth of species name, aureus, or “golden,” refers to the typical creamy
P.  acnes. Other medications such as isotretinoin (Accutane) act color of the S. aureus colonies. This bacterium is an extremely
by reducing sebum production. Accutane is generally reserved for important potential pathogen and is mentioned frequently
the most serious cases of acne because it has potentially serious throughout this text as the cause of a number of medical condi-
side effects. Squeezing acne lesions is ill-advised, because it may tions (table 22.2).
cause the inflamed follicles to rupture, leading to more acne scars. One of the most useful identifying characteristics of S. aureus
It can also introduce bacteria into the bloodstream, which could that distinguishes it from most other staphylococci is that it pro-
spread the infection to multiple locations throughout the body. duces coagulase and clumping factor. Despite the -ase ending,
coagulase is not an enzyme, but a secreted protein. Clumping
factor is a functionally similar, but genetically distinct, protein.
Hair Follicle Infections It is easy to test for in the laboratory and it is often called “slide
Hair follicle infections are generally mild and commonly clear up coagulase.” Both coagulase and clumping factor are important
without treatment. In some instances, however, they progress into virulence factors for S. aureus. Other staphylococcal species, such
severe or even life-threatening disease. as Staphylococcus epidermidis, cause disease infrequently and
lack the genes and the proteins for coagulase and clumping factor.
Signs and Symptoms virulence factor, p. 383
There are three outcomes of hair follicle infections, listed
by increasing severity: folliculitis, furuncles, and carbuncles. Pathogenesis
Folliculitis is inflammation of one or more hair follicles, causing Infection begins when Staphylococcus aureus attaches to the
small red bumps, or pimples. The hair can be pulled from its fol- cells of a hair follicle, multiplies, and spreads to involve the seba-
licle, releasing a small amount of pus. The infection often goes ceous glands. The infection induces an inflammatory response
away without further treatment. If the infection extends from the with swelling and redness, followed by attraction and accu-
follicle to adjacent tissues, it will cause a localized redness, swell- mulation of neutrophils. If the infection continues, the follicle
ing, severe tenderness, and pain. This lesion is called a furuncle becomes a plug of inflammatory cells and dead tissue overlying
or boil. Pus may drain from the boil along with a plug of inflam- a small abscess (figure 22.2). The infectious process sometimes
matory cells and dead tissue. A furuncle may worsen to form a spreads deeper, reaching the subcutaneous tissue where a large
carbuncle, a large area of redness, swelling, and pain with several abscess forms. This subcutaneous abscess is responsible for the
sites of draining pus. Carbuncles usually develop in areas of the painful localized swelling called a boil. Without effective treat-
body where the skin is thick, such as the back of the neck. Fever ment, pressure within the abscess may increase, causing it to
is often present, along with other signs of a serious infection. expand to other hair follicles and form a carbuncle. If organisms
fever, p. 351 enter the bloodstream, the infection can spread to other parts of
the body, such as the heart, bones, or brain.
Causative Agent S. aureus strains can have many different virulence fac-
Most furuncles and carbuncles, as well as many cases of fol- tors, although not all pathogenic strains make the same factors
liculitis, are caused by Staphylococcus aureus. The name derives (table 22.3). Nearly all strains have protein A, a cell wall com-
from the Greek root staphyle or “a bunch of grapes,” referring ponent that interferes with phagocytosis (see figure  16.11).
 Part IV Infectious Diseases 525

Staphylococcus aureus
infects hair follicle. Hair

Epidermis

Sebaceous
gland
Dermis
White
blood cell
Blood
Subcutaneous vessel
tissue

Plug of pus

Infection spreads to
subcutaneous tissue. Abscess

Accumulation of
white blood cells

FIGURE 22.2 Pathogenesis of a Boil (Furuncle) Staphylococcus aureus infects a hair follicle through its opening on the
skin surface. The infection produces a plug of necrotic material, a small abscess in the dermis, and finally, a larger abscess in
the subcutaneous tissue.
? Why is it a bad idea to squeeze a pimple?

TABLE 22.3 Properties of Staphylococcus aureus Implicated in Its Virulence

Product Effect

Capsule Inhibits phagocytosis

Clumping factor Attaches the bacterium to fibrin, fibrinogen, and plastic devices
Coagulase May slow progress of leukocytes into infected area by producing clots in the surrounding capillaries
Enterotoxins Superantigens cause food poisoning if ingested, cause toxic shock if systemic
Exfoliatin Separates layers of epidermis, causing scalded skin syndrome
Fibronectin-binding protein Attaches bacterium to acellular tissue substances, endothelium, epithelium, clots, indwelling plastic devices
Hyaluronidase Breaks down hyaluronic acid component of tissue, thereby allowing infection to spread
Leukocidin Kills neutrophils or causes them to release their enzymes
Lipase Breaks down fats by hydrolyzing the bond between glycerol and fatty acids
Proteases Degrade collagen and other tissue proteins
Protein A Binds to Fc portion of antibody, thereby interfering with opsonization
Toxic shock syndrome toxin Causes rash, diarrhea, and shock
α-toxin Makes holes in host cell membranes
526 Chapter 22 Skin Infections

Clumping factor Fibrinogen FIGURE 22.3 Virulence Molecules

Lipoteichoic acid on the Staphylococcus aureus Cell
Envelope A polysaccharide capsule,
Antibody protein A, and clumping factor
contribute to the virulence of this
Protein A organism.
? How does protein A help
Staphylococcus aureus evade
phagocytes?

Polysaccharide
capsule

Peptidoglycan

Staphylococci
Cytoplasmic
membrane

Many strains of S. aureus also synthesize a polysaccharide capsule kills host cells by making holes in their membranes. Leukocidins
that inhibits phagocytosis (figure 22.3). protein A, p. 390 kill white blood cells. A relatively small percentage of S. aureus
Coagulase and clumping factor allow S. aureus to colonize strains produce one or more additional disease-specific toxins.
soft tissues while avoiding the immune response. Coagulase is an One of these, exfoliatin, causes staphylococcal scalded skin syn-
important virulence factor because it activates a blood protein called drome, the next disease described in this chapter. membrane-
prothrombin to form thrombin. Thrombin then converts fibrinogen damaging toxin, p. 392
to fibrin, the fibrous protein that forms the weblike foundation
of blood clots. While most coagulase is secreted, some is tightly Epidemiology
bound to the bacterial cells and coats their surface with fibrin when Staphylococcus aureus can be found in the nostrils of virtually
they come into contact with blood. This helps disguise the bacteria, everyone at one time or another. About 20% of healthy adults have
hiding them from the immune system. Fibrin-coated staphylo- continually positive nasal cultures for a year or more, whereas
cocci resist phagocytosis. Clumping factor is a virulence factor over 60% will be colonized at some time during a given year. The
for S. aureus because it attaches to fibrinogen and fibrin present in organisms are mainly spread to other parts of the body and to the
damaged tissues, and helps the bacteria colonize wound surfaces. environment by the hands. Although the nostrils seem to be the pre-
Because plastic devices such as intravenous catheters and heart ferred habitat of S. aureus, moist areas of skin are also frequently
valves quickly become coated with fibrinogen after insertion into colonized. People with boils and other staphylococcal infec-
the bloodstream, they also become targets for colonization. tions shed large numbers of S. aureus and should not work with
S. aureus secretes many enzymes that can damage host tis- food, or near patients with surgical wounds or chronic illnesses.
sues. Hyaluronidase degrades hyaluronic acid, a component of Staphylococci survive well in the environment, so they are easily
host tissue that helps hold the cells together. Proteases degrade transferred from one host to another via fomites. fomites, p. 440
various host proteins including collagen, the white fibrous protein Because S. aureus is so common, epidemics of staphylo-
found in skin, tendons, and connective tissue. Lipases, which coccal disease can generally be traced to their sources only by
degrade lipids, provide a food source when S. aureus colonizes identification of the epidemic strain. To precisely identify an epi-
oily hair follicles. demic strain, techniques such as molecular typing, phage typing,
Most strains of S. aureus also produce one or more toxins and antibiogram determination must be used (see section 10.4).
that damage or kill host cells. The membrane-damaging α-toxin characterizing strain differences, p. 247
 Part IV Infectious Diseases 527

Treatment and Prevention MicroByte

Treatment of boils and carbuncles may require minor surgery to Individuals colonized with S. aureus may have as many as 108 cells
of the bacterium per nostril.
drain the pus from the lesion. Afterward, patients are usually given
a course of oral antibiotics.
Treating S. aureus infections can be complicated because
many strains are resistant to multiple antibacterial medications. As Staphylococcal Scalded Skin Syndrome
described in chapter 20, the organism is an important example of Staphylococcal scalded skin syndrome (SSSS) is a potentially fatal,
emerging antibiotic resistance. When penicillin was first intro- toxin-mediated disease. This disease occurs mainly in infants.
duced, nearly all strains were susceptible. However, strains that
produce a penicillinase (a type of β-lactamase) spread quickly, so Signs and Symptoms
that most isolates are now resistant to this medication. Strains As the name suggests, staphylococcal scalded skin syn-
producing modified versions of penicillin-binding proteins also drome causes a patient’s skin to appear burned (figure 22.4).
appeared. These strains, referred to as MRSA (methicillin- It begins as a generalized redness of the skin, sometimes
resistant Staphylococcus aureus) are resistant to all β-lactam affecting the entire body. Other signs and symptoms include
medications, including penicillin derivatives and cephalosporins. malaise—a vague feeling of discomfort and uneasiness—
Many MRSA strains are resistant to a number of other medica- irritability, and fever. The nose, mouth, and genitalia may be
tions as well. Before 1997, even the most resistant examples were painful for one or more days before the typical features of the
reliably treated with vancomycin. Since then, however, the first disease become apparent. Within 48 hours after the redness
vancomycin-intermediate S. aureus (VISA) and vancomycin- appears, the skin becomes wrinkled, and large blisters filled
resistant S. aureus (VRSA) strains were identified. Several medi- with clear fluid develop. The skin is tender to the touch, peels
cations active against VRSA have been marketed, including easily, and feels like sandpaper.
linezolid, daptomycin, and tigecycline. Linezolid represents a new
class of antibacterials, the oxazolidinones. Hopefully, develop- Causative Agent
ment of new medications will keep pace with the growth of
SSSS is caused by Staphylococcus aureus strains that produce a
bacterial resistance, but this will depend on humans avoiding
toxin called exfoliatin. Only about 5% of S. aureus strains pro-
overuse of these valuable substances.  β-lactamase, p. 464 VISA
duce this toxin, which destroys material that binds together the
and VRSA, p. 473
outer layers of epidermis. At least two kinds of exfoliatins exist:
When they first appeared, most MRSA strains could be
One is coded for by a plasmid gene, and the other by a chromo-
traced to hospitals and clinics. More recently however, completely
somal gene. plasmids, p. 208
different strains have become widespread among healthy carri-
ers in the community. These new strains are called CA-MRSA
(community-acquired MRSA) to distinguish them from hospital-
acquired strains (HA-MRSA). The most common CA-MRSA
strain in the United States is USA-300. All current isolates are
thought to be the progeny of one original population. Generally,
these CA-MRSA strains are resistant only to methicillin and other
β-lactam antibiotics, and macrolides. They are highly virulent and
possess a group of genes that codes for a leukocyte-destroying
leukocidin. There is some debate as to whether this leukocidin
contributes to higher morbidity and mortality or if it is simply a
trait shared by most strains of CA-MRSA. penicillins and other
β-lactam antibiotics, p. 463 macrolides, p. 466
In an effort to control the spread of MRSA, many hospitals
screen patients at admission. The patients who carry a MRSA
strain are isolated and given appropriate antibacterial treatment to
limit spread of the bacteria to other patients or staff. Some hospi-
tals also screen patients for MRSA when they are discharged from
the hospital to make sure they do not take a MRSA strain home
with them. carriers, p. 439
Prevention of staphylococcal skin disease is very difficult
because so many individuals carry the organism. Applying an
antibacterial cream to the nostrils and using soaps containing
an antibacterial agent such as hexachlorophene to wash the skin
reduce the bacterial burden and may eliminate the carrier state. FIGURE 22.4 Staphylococcal Scalded Skin Syndrome
Because S. aureus is most commonly transmitted by the hands, (SSSS) A toxin called exfoliatin, produced by certain strains of
handwashing and regular use of hand sanitizers are both effective Staphylococcus aureus, causes the outer layer of skin to separate.
means of limiting the spread of these bacteria. ? Why is SSSS sometimes fatal?
528 Chapter 22 Skin Infections

Pathogenesis
Exfoliatin causes the epidermis to split just below the dead kerati-
nized outer layer. The S. aureus strains that cause SSSS generally
grow in a relatively small area. However, the exfoliatin produced
by the colonizing bacteria is carried through the bloodstream
to damage large areas of skin. Because the outer layers of skin
are lost (as happens with a severe burn), there is significant loss
of body fluid and danger of secondary infection with Gram-
negative bacteria such as Pseudomonas sp. secondary infection,
p. 382 Pseudomonas sp., p. 265

Epidemiology
Staphylococcal scalded skin syndrome can appear in any age
FIGURE 22.5 Impetigo This type of pyoderma is often caused by
group but occurs most frequently in newborn infants. Elderly
Streptococcus pyogenes and may result in glomerulonephritis.
and immunocompromised individuals are also at increased risk.
Transmission is generally from person to person. The disease usu- ? How is impetigo spread in a population?
ally appears in isolated cases, although small epidemics in nurser-
ies sometimes occur.
Causative Agent
Treatment and Prevention Many cases of impetigo, even epidemics, are due to Streptococcus
Initial therapy for staphylococcal scalded skin syndrome includes pyogenes. These Gram-positive, chain-forming organisms are
an antibiotic such as methicillin. As with severe burns, dead skin is group A streptococci, bacteria characterized by the presence of
removed to help prevent secondary infection. Although the disease a polysaccharide called group A carbohydrate in their cell walls.
can be fatal, prompt therapy usually leads to full recovery. There are Like Staphylococcus aureus, S. pyogenes causes a variety of
no preventive measures except to place patients suspected of having different diseases and is mentioned in numerous places through-
SSSS in protective isolation. This limits the spread of the pathogen out this text. The most detailed description is in chapter 21.
to others and helps prevent secondary infections in the isolated S. aureus also causes impetigo. Table 22.5 compares S. aureus and
patient. Table 22.4 describes the main features of this disease. S. pyogenes. hemolysis, p. 96 Streptococcus pyogenes, p. 487

Pathogenesis
Streptococcal Impetigo
Many different strains of S. pyogenes exist, some of which can
Impetigo is the most common type of pyoderma, a skin infec- colonize the skin. Minor injuries introduce the bacteria into the
tion characterized by pus production (figure  22.5). Pyodermas deeper layer of epidermis, often leading to an infection. Scratching
can result from infection of an insect bite, burn, scrape, or other an infected area may spread the bacteria to other areas of the skin.
wound. Sometimes, the injury is so slight it is not apparent. The surface components of S. pyogenes, notably a hyaluronic
acid capsule and a cell wall component called M protein, interfere
Signs and Symptoms with phagocytosis (see figure 21.4). A number of extracellular
Impetigo is a superficial skin infection, causing inflammation in products contribute to the virulence of S. pyogenes, similar to
patches of epidermis just beneath the dead, scaly outer layer. Thin- those described for S. aureus. These include enzymes such as
walled blisters develop, then break, and are replaced by oozing, proteases, nucleases, and hyaluronidase. None of them appears
yellowish crusts of drying plasma. Usually, patients experience to be essential, however, because antibodies against them fail to
little fever or pain, but lymph nodes near the involved areas often protect experimental animals. A more detailed description of these
enlarge, indicating that bacterial products have entered the lym- virulence factors is in chapter 21. virulence factors of S. pyogenes,
phatic system and an immune response is occurring. plasma, p. 426 p. 488 M protein, p. 390

TABLE 22.4 Staphylococcal Scalded Skin Syndrome

Signs and symptoms Sensitive red rash with sandpaper texture, malaise, irritability, fever, large blisters, peeling of skin
Incubation period Variable, usually days
Causative agent Strains of Staphylococcus aureus that produce exfoliatin toxin
Pathogenesis Exfoliatin is produced by organisms at an infection site, usually of the skin, and carried by the bloodstream to the
epidermis, where it causes a split in a cellular layer; loss of body fluid and secondary infections contribute to mortality.
Epidemiology Person-to-person transmission; seen mainly in infants, but can occur at any age
Treatment Treatment with penicillinase-resistant penicillins; removal of dead tissue. Isolate the patient to prevent secondary
and prevention infection and to limit the spread of the disease to others.
 Part IV Infectious Diseases 529

TABLE 22.5 Streptococcus pyogenes Versus Staphylococcus aureus

Streptococcus pyogenes Staphylococcus aureus
Morphology Chains of Gram-positive cocci Clusters of Gram-positive cocci
Growth Beta-hemolytic colonies; catalase negative, coagulase Golden, hemolytic colonies, catalase-negative,
characteristics negative, obligate fermenter coagulase-positive, facultative anaerobe
Virulence Cell wall contains group A polysaccharide, an Fc Cell wall contains an Fc receptor (protein A). Bacterium
factors receptor (protein G), and M protein. Bacterium produces produces superantigens, hemolysins, leukocidin,
hemolysins (streptolysins O and S), streptokinase, DNase, hyaluronidase, nuclease, protease, and others
hyaluronidase, and others
Diseases Impetigo, strep throat, wound infections, scarlet fever, Boils, staphylococcal scalded skin syndrome, wound
puerperal fever, toxic shock, and necrotizing fasciitis. infections, abscesses, bone infections, impetigo, food
Complications: glomerulonephritis and rheumatic fever poisoning, and staphylococcal toxic shock syndrome

Even though impetigo is limited to the epidermis, strepto- Signs and Symptoms
coccal products are absorbed into the circulation. The immune Rocky Mountain spotted fever begins suddenly with a headache,
response to these proteins is thought to cause post-streptococcal pains in the muscles and joints, and fever. Within a few days of
sequelae, particularly acute glomerulonephritis. post-streptococcal infection, a rash characterized by faint pink spots appears on the
sequelae, p. 489 glomerulonephritis, p. 490 palms, wrists, ankles, and soles. This rash, caused by blood leak-
ing from damaged vessels, spreads up the arms and legs to the rest
Epidemiology of the body (figure 22.6). Bleeding may occur at other sites, such
Impetigo is most common among poor young children living in hot,
humid areas. Person-to-person contact spreads the disease, as do
insects, and fomites such as toys and towels. Impetigo patients often
become throat and nasal carriers of S. pyogenes. fomites, p. 440

Treatment and Prevention

So far, S. pyogenes strains remain susceptible to penicillin. For
patients allergic to penicillin, erythromycin can be substituted.
People with impetigo should avoid contact with others to pre-
vent spreading the bacteria. The transmission of impetigo can be
limited by keeping skin clean and avoiding sharing personal items
such as towels and washrags. Antiseptics and wound cleansing
also decrease the chance of infection. Table 22.6 summarizes the
main features of impetigo.

Rocky Mountain Spotted Fever

Rocky Mountain spotted fever (RMSF) was first recognized in
the Rocky Mountain area of the United States—thus its name. FIGURE 22.6 Rash Caused by Rocky Mountain Spotted
The disease is caused by an obligate intracellular pathogen called Fever Characteristically, the rash begins on the arms and legs,
rickettsia and is transmitted by certain species of ticks, mites, and spreads centrally, and as shown in this photo, becomes hemorrhagic.
lice. the genus Rickettsia, p. 277 ? What processes cause this rash?

TABLE 22.6 Impetigo

Signs and symptoms Blisters that break, releasing plasma and pus; formation of golden-colored crusts; lymph node enlargement
Incubation period 2 to 5 days
Causative organisms Streptococcus pyogenes, Staphylococcus aureus
Pathogenesis Organisms entering the skin through minor breaks; certain strains of S. pyogenes that cause impetigo can also
cause glomerulonephritis
Epidemiology Spread by direct contact with carriers or patients with impetigo, insects, and fomites
Treatment and prevention An oral penicillin if cause is known to be S. pyogenes; otherwise, an anti-staphylococcal antibiotic orally or
topically. Cleanliness; care of skin injuries.
530 Chapter 22 Skin Infections

Rodent cell them, because endothelial cells are not normally capable of phago-
cytosis. Once inside the host cell, the bacteria leave the phagosome
and multiply in both the cytoplasm and nucleus. They coat them-
selves in host cell actin and use this protein to move into adjacent
host cells. Eventually, the host cell membrane is so damaged by
this process that the cell ruptures, releasing the rickettsias. These
travel through the bloodstream, infecting even more cells.
Rodent Infection can also extend into the walls of the small blood ves-
cell nucleus sels, causing an inflammatory reaction. This leads to clotting in the
blood vessels, and small areas of necrosis (tissue death). This pro-
cess causes the hemorrhagic skin rash. More ominously, clotting
occurs throughout the body, resulting in damage to vital organs
Rickettsias such as the kidneys and heart. Potentially even more serious is
the release of endotoxin (lipopolysaccharide) into the bloodstream
from the rickettsial cell walls, causing shock and generalized
10 μm bleeding because of disseminated intravascular coagulation.
FIGURE 22.7 The Obligate Intracellular Bacterium Rickettsia lipopolysaccharide, p. 60 disseminated intravascular coagulation, p. 674
rickettsii Growing in a Rodent Cell
? Do you think Rickettsia rickettsii can grow on blood agar? Epidemiology
Rocky Mountain spotted fever occurs in a sporadic distribution
throughout the Americas. Despite the name of the disease, the
as the nose and mouth. Damage to the heart, kidneys, and other highest incidence in the United States has generally been in the
body tissues can cause a drop in blood pressure, shock, and death south Atlantic and south-central states (figure 22.8).
unless treatment is given promptly. RMSF is a zoonosis, maintained in nature in various species
of ticks and mammals. Rickettsias generally do not cause disease
Causative Agent in their natural hosts—the host and pathogen co-evolved to reach a
Rocky Mountain spotted fever is caused by Rickettsia rickettsii state of balanced pathogenicity. Humans, being an accidental host,
(figure 22.7). The organisms are tiny, Gram-negative, non-motile often develop severe disease. balanced pathogenicity, p. 385
coccobacilli that require eukaryotic cells to grow. This obligate Several species of ticks transmit RMSF to humans. The main
intracellular growth makes the bacteria very difficult to grow in vector in the western United States is the wood tick, Dermacentor
culture. R. rickettsii can sometimes be identified early in an infec- andersoni (figure  22.9), while in the East it is the dog tick,
tion by visualizing the organisms in skin lesion biopsies (pieces of D.  variabilis. Once infected, ticks remain so for life, transmitting
tissue surgically removed from the lesion site). Also, their DNA R. rickettsii from one generation to the next through their
can be amplified by the polymerase chain reaction (PCR) and eggs. Ticks are most active from April to September. Not sur-
identified with a probe. PCR, p. 227 probe, p. 232 prisingly, most cases of RMSF occur during this time of year.
zoonoses, p. 439
Pathogenesis
The bite of a tick infected with R. rickettsii
transmits Rocky Mountain spotted fever.
The tick remains attached for hours while
it feeds, but the process is usually painless
and goes unnoticed. The pathogen is not
transmitted until a tick has fed from 4 to
10 hours. The bacteria are released into
capillary blood with the tick saliva and are
taken up by the endothelial cells lining the
small blood vessels. There is evidence that
R. rickettsii force the endothelial cells to engulf

0
1–14 cases (on average)
≥15

FIGURE 22.8 Rocky Mountain Spotted

Fever Number of reported cases, by
county—United States, 2005.
? Why do most cases of RMSF occur in the
summer months?
 Part IV Infectious Diseases 531

FIGURE 22.10
Erythema Migrans,
the Characteristic
Rash of Lyme
Disease The rash
usually has a targetlike
or bull’s-eye appear-
ance. Although highly
suggestive of Lyme
disease, many people
with the disease do
not develop the rash.
? What causes the
bull’s-eye pattern
of this rash?

FIGURE 22.9 Dermacentor andersoni, the Wood Tick

The wood tick is the main vector of Rocky Mountain spotted fever
in the western United States.
? What can be done to limit exposure to these ticks?

Treatment and Prevention Lyme Disease

The antibiotics doxycycline and chloramphenicol are very effec- Lyme disease was first recognized as a distinct entity in the mid-
tive in treating Rocky Mountain spotted fever if given early in the 1970s, when a cluster of cases occurred in Lyme, Connecticut. It
disease, before irreversible damage to vital organs has occurred. was not until 1982 that the causative agent was first identified in
Without treatment, the overall case-fatality rate from the disease ticks by Dr. Willy Burgdorfer at the Rocky Mountain Laboratories
is about 20%, but it can be much higher in elderly patients. With in Montana. We now know that Lyme disease was widespread
early diagnosis and treatment, the case-fatality rate is reduced to long before its recognition at Lyme. The ecology of the disease
less than 5%. is complex and still incompletely understood, but we are begin-
No vaccine against RMSF is currently available to the pub- ning to get some answers as to why the disease has increased and
lic. The disease can be prevented if people take precautions to extended its range. About 25,000 new cases occur each year, mak-
(1)  avoid tick-infested areas when possible; (2) wear protective ing it the most common vector-borne disease in the United States.
clothing; (3) use tick repellents such as DEET (N,N-diethyl-meta-
toluamide); (4) carefully inspect their bodies (especially the scalp, Signs and Symptoms
armpits, and groin) for ticks several times daily; and (5) remove
Signs and symptoms of Lyme disease can be divided roughly into
attached ticks carefully to avoid crushing them and thereby con-
three stages, although individual patients may be asymptomatic in
taminating the bite wound with their infected tissue fluids. To
one or more of these:
remove ticks, blunt tweezers can be used to grasp the tick by its
mouthparts and then gently pull it away from the skin. The site ■ Early localized infection. This stage typically begins a few
of the bite should be treated with an antiseptic. Touching the tick days to several weeks after a bite by an infected tick. It is
with a hot object, gasoline, or whiskey is ineffective. The main characterized by enlargement of nearby lymph nodes and a
features of Rocky Mountain spotted fever are summarized in circular skin rash called erythema migrans (figure  22.10).
table 22.7. The rash begins as a red spot or bump at the site of the tick

TABLE 22.7 Rocky Mountain Spotted Fever

Signs and symptoms Headache, pains in muscles and joints, and fever, followed by a hemorrhagic rash that begins on the extremities
Incubation period 4 to 8 days
Causative organism Rickettsia rickettsii, an obligate intracellular bacterium
Pathogenesis Organisms multiply at site of tick bite; invade the bloodstream and then infect endothelial cells; vascular lesions
and immune response to endotoxin account for pathologic changes.
Epidemiology A zoonosis transmitted by bite of infected tick, usually Dermacentor sp.
Treatment Treated with: doxycycline or chloramphenicol. Prevented by avoiding tick-infested areas, using tick repellent,
and prevention removal of ticks within 4 hours.
532 Chapter 22 Skin Infections

■ Late persistent infection. This begins around 6 months

after the skin rash. This stage is characterized by joint pain
and swelling, usually of large joints such as the knee. These
signs and symptoms slowly disappear over subsequent years.
Chronic nervous system impairments such as localized pain,
paralysis, and depression can occur.

Causative Agent
Lyme disease is caused by Borrelia burgdorferi, a large, Gram-
negative, microaerophilic spirochete (figure 22.11). The Borrelia
10 μm chromosome is unusual in that it is linear and present in multi-
ple copies. B. burgdorferi also contains numerous circular and
FIGURE 22.11 Scanning Electron Micrograph of Borrelia
burgdorferi, the Cause of Lyme Disease linear plasmids that contain genes usually found on bacterial
chromosomes. Studying this unusual bacterium may lead to an
? How would you describe the shape of this organism?
understanding of how it can infect such widely differing species
as mice, lizards, and ticks. spirochetes, p. 272 microaerophilic, p. 90

bite and slowly enlarges to eventually reach a diameter Pathogenesis

of about 15 cm (6 inches). The advancing edge is bright The spirochetes are introduced into the skin by the bite of an
red, leaving behind an area of fading redness as the lesion infected tick. They multiply and migrate outward in a circular
enlarges. This characteristic bull’s-eye rash is the hallmark of fashion. The lipopolysaccharide layer of these Gram-negative
Lyme disease but is not present in all cases. Most of the other bacteria causes an inflammatory reaction in the skin, producing
signs and symptoms that occur during this stage are flulike— the expanding rash. The host’s immune response is initially sup-
malaise, chills, fever, headache, stiff neck, joint and muscle pressed, allowing continued multiplication of the spirochete at
pains, and backache. the site. The organisms then enter the bloodstream and spread to
■ Early disseminated infection. This generally begins 2 to 8 all parts of the body. This dissemination accounts for the flulike
weeks after the first signs and symptoms appear, and affects signs and symptoms of the first stage. At this point, an intense
the nervous system. Electrical conduction within the heart immune response occurs, but after the first few weeks, it becomes
is impaired, leading to dizzy spells or fainting. A temporary very difficult to recover B. burgdorferi from blood or body tis-
pacemaker may be required to maintain a normal heartbeat. sues. The immune response against the bacterial antigens is prob-
Nervous system effects also include one or more of the fol- ably responsible for the signs and symptoms of the second and
lowing: paralysis of the face, severe headache, pain when third stages. The third stage of Lyme disease is characterized by
moving the eyes, difficulty concentrating, emotional instabil- arthritis, and the affected joints have high concentrations of reac-
ity, fatigue, and impairment of the nerves of the legs or arms. tive immune cells and immune complexes. Evidence suggests a
role for autoimmune response against host tissues in some cases.
autoimmunity, p. 412 lipopolysaccharide, p. 60

Epidemiology
Lyme disease is a zoonosis with humans
being an accidental host. It is widespread
in the United States, and its incidence
depends on complex ecological factors
(figure 22.12). Several species of ticks have
been implicated as vectors, but the most
important in the eastern United States is the black-
legged (deer) tick, Ixodes scapularis (figure 22.13).
In some areas of the East Coast, 80% of these ticks

0
1–14 cases (on average)
≥15

FIGURE 22.12 Number of Reported Lyme Disease

Cases—2005
? Why is the distribution of Lyme disease cases not uniform
across the United States?
 Part IV Infectious Diseases 533

Female drops from host

and lays uninfected eggs.

Spring

Eggs

Uninfected
larvae hatch. Summer

First year
Larvae feed on
infected animal
2 mm
host and
acquire
Borrelia Fall
FIGURE 22.13 The Black-Legged (Deer) Tick, Ixodes burgdorferi.
scapularis, Adult and Nymph This tick is the most important
vector of Lyme disease in the eastern and north-central United States.
? Why are people not always aware that they have been bitten by Infected
this tick? larvae
become dormant.
Winter
Infected larvae develop
into nymphs.

are infected with B. burgdorferi. Because of the tick’s small

Infected
size, they often feed and drop off their host without being nymphs feed,
detected—two-thirds of Lyme disease patients are unable to transmitting
Borrelia Spring
recall a tick bite.
burgdorferi.
The ticks mature during a 2-year cycle (figure  22.14). A
larval form emerges from the egg. After growing, it molts, shed-
ding its outer covering to become a nymph. After another molt,
the nymph becomes the sexually mature adult form. The nymph
actively seeks blood meals and is therefore mainly responsible Infected
nymphs
for transmitting Lyme disease. B. burgdorferi is easily transferred Summer
mature into
from the tick to its preferred host, the white-footed mouse. Once adults.

Second year
infected, the mouse develops a sustained bacteremia and becomes
a source of infection for other ticks. The spirochete is rarely
passed from adult tick to its offspring. bacteremia, p. 384
Adults feed on animal
Infected ticks and mice are the main reservoirs of B. burgdorferi, (deer) host and mate.
but deer play an important role as well. They are the preferred host Fall
of the adult ticks and the site where tick-mating occurs. Moreover,
deer can quickly spread the disease over a wide area. Tick nymphs
are most active from May to September, corresponding to the
peak occurrence of Lyme disease. Adult ticks sometimes bite
humans late in the season and transmit the disease. Infectious ticks
can be present in well-mowed lawns as well as in wooded areas.
Winter
reservoirs, p. 438
Female dormant;
male dies.
Treatment and Prevention
Several antibiotics are effective in patients with early stages of
Lyme disease. In late disease, antibiotic treatment is less effective, FIGURE 22.14 Life Cycle of the Black-Legged (Deer) Tick,
most likely because the spirochetes are not actively multiplying Ixodes scapularis The life cycle covers 2 years, during which the
tick obtains three blood meals. The males die soon after mating, the
and antibacterial medications usually target dividing bacteria. females after depositing their eggs in the following spring. Variations
Nevertheless, prolonged treatment with intravenous ampicillin or in the life cycle occur, probably dependent on climate and food
ceftriaxone has cured many cases. availability for the natural hosts.
General preventive measures for Lyme disease are the same ? What time of the year has the highest number of new cases of
as those for Rocky Mountain spotted fever and include avoiding Lyme disease?
534 Chapter 22 Skin Infections

TABLE 22.8 Lyme Disease

1 Bite of tick infected with Signs and Early localized infection: Enlarging
Borrelia burgdorferi introduces symptoms rash at the site of the bite; lymph
the bacteria into the skin. node enlargement near bite, flulike
2 B. burgdorferi reproduce and symptoms. Early disseminated
spread radially in the skin, infection: Heart and nervous system
causing an expanding red rash involvement. Late persistent infection:
which tends to clear centrally. Chronic arthritis and nervous system
impairment.
3 The bacteria enter the
bloodstream, cause fever, Incubation Approximately 1 week
acute injury to the heart and 3 period
nervous system. Causative Borrelia burgdorferi, a spirochete
4 Chronic symptoms develop, agent
such as arthritis and paralysis 4
1 Pathogenesis Spirochetes injected into the skin
due to persisting bacteria and by an infected tick multiply and
the immune response to them. 2 spread radially; the spirochetes enter
No person-to-person the bloodstream and are carried
transmission. throughout the body; the immune
reaction to bacterial antigen causes
tissue damage.
Epidemiology Spread by the bite of ticks, Ixodes
sp., usually found in association
with animals such as white-footed
mice and white-tailed deer living in
wooded areas.
Treatment and Early treatment with doxycycline and
prevention others; prolonged antibiotic therapy
in chronic cases. Protective clothing;
tick repellents.

exposure to ticks. There is no available vaccine. Table 22.8 sum- Several different viruses initially infect the upper respiratory tract,
marizes some features of Lyme disease. but are carried in the blood to the skin where they cause distinctive
skin rashes. These diseases are usually diagnosed based on the
MicroAssessment 22.2 appearance of the rash and other clinical findings. When the dis-
ease is not typical, however, immunological testing can be used to
Folliculitis, furuncles, and carbuncles are usually caused by
Staphylococcus aureus. Some strains of S. aureus produce exfoliatin identify antibodies against each virus. Other viruses enter through
and can cause staphylococcal scalded skin syndrome. Impetigo is a skin lesions (see Perspective 22.1). immunological testing, p. 426
kind of pyoderma that is often caused by Streptococcus pyogenes.
Rocky Mountain spotted fever is caused by Rickettsia rickettsii and is Varicella (Chickenpox)
transmitted by ticks. Lyme disease is caused by Borrelia burgdorferi
and is also transmitted by ticks. It is the most common vector-borne Chickenpox is the popular name for varicella, a rash that was
disease in the United States. common in childhood before the varicella vaccine was introduced
4. List four extracellular products of Staphylococcus aureus that in 1995. The causative agent is a member of the herpesvirus fam-
contribute to its virulence. ily. All herpesviruses produce latent infections that can reactivate
5. Describe the characteristic rash of Lyme disease. long after recovery from the initial illness. latent infections, p. 322

6. Patients with staphylococcal scalded skin syndrome often do

not have Staphylococcus aureus growing in the affected areas. Signs and Symptoms
Why do you think this is the case? + Most cases of childhood chickenpox are mild, and sometimes go
unnoticed. The typical case begins as small, red spots called mac-
ules, progressing to little bumps called papules, and then to small
22.3 ■ Skin Diseases blisters called vesicles. The lesions appear at different times, so
Caused by Viruses that any time during the rash, they are at various stages of devel-
opment from macule to papule or vesicle (figure  22.15). They
Learning Outcomes can occur anywhere on the body, but usually first appear on the
6. Compare and contrast chickenpox, measles, and rubella. back of the head, then the face, mouth, main body, and arms and
7. Compare and contrast fifth disease and roseola. legs. Patients can have only a few lesions or many hundreds. The
8. Describe the process that leads to warts. lesions are pruritic (itchy) and scratching them can lead to serious,
even fatal, secondary infection by S. pyogenes or S. aureus.
 Part IV Infectious Diseases 535

FIGURE 22.15
A Child with
Chickenpox
(Varicella)
Characteristically,
lesions in various
stages of evolution—
macules, papules,
vesicles, and
pustules—are present.
? Is chickenpox
caused by a
poxvirus?

FIGURE 22.16 Shingles (Herpes Zoster) The rash mimics that of

chickenpox, except that it is limited to a sensory nerve distribution on
The signs and symptoms of chickenpox tend to be more one side of the body.
severe in older children and adults. About 20% of adults develop ? Can you catch chickenpox or shingles from a person with shingles?
pneumonia. This may lead to rapid breathing, cough, shortness of
breath, and a dusky skin color from lack of O2. The pneumonia
subsides with the rash, but respiratory signs and symptoms often
MicroByte
persist for weeks. Chickenpox is a threat to immunocompromised
Non-immune people can contract chickenpox from someone with
patients of any age—the virus damages the lungs, heart, liver, shingles.
kidneys, and brain, resulting in death in about 20% of the cases.
Chickenpox is a major threat to babies. It has a case-fatality
rate as high as 30% in newborn infants. Babies born to mothers Causative Agent
who had chickenpox early in pregnancy occasionally develop Chickenpox is caused by varicella-zoster virus (VZV), a member
congenital varicella syndrome. These babies may be born with of the herpesvirus family. It is an enveloped, double-stranded
underdeveloped head and limbs, and cataracts. DNA virus, indistinguishable from other herpes-
Reactivation of the chickenpox virus results in shingles. This viruses in appearance (figure 22.17).
disease can occur at any age but becomes increasingly common
later in life. It begins with pain in the area supplied by a sensory
nerve, often on the chest or abdomen but sometimes on the face
or an arm or leg. After a few days to 2 weeks, a rash characteristic
of chickenpox appears, but unlike chickenpox, the rash is usually
restricted to the area supplied by the branches of the involved
sensory nerve (figure 22.16). The rash generally subsides within
a week, but pain may persist for weeks, months, or longer. In
people with AIDS or other serious immunodeficiency, the rash
often spreads to involve the entire body, as in a severe case of
chickenpox. Shingles does not spread from person to person, since
it is caused by a reactivation of latent virus.
A curious, rare affliction known as Reye’s syndrome occa-
sionally occurs in association with chickenpox and a number of
other viral infections, usually within 2 to 12 days of the onset of
the infection. The patient begins vomiting and then slips into a
coma. The syndrome occurs predominantly in children between
5 and 15 years old and is characterized by liver and brain damage 100 nm

and a death rate as high as 30%. Epidemiologic evidence suggests FIGURE 22.17 Varicella-Zoster Virus Electron micrograph of the
that aspirin therapy increases the risk of Reye’s syndrome and viral cause of chickenpox and shingles.
should be avoided in children with fever. ? How does this virus cause both shingles and chickenpox?
536 Chapter 22 Skin Infections

PERSPECTIVE 22.1
The Ghost of Smallpox: An Evil Shade
Historically, smallpox epidemics have been respiratory secretions, but also by antibody response occurs rapidly and
devastating to the Americas. In the 1500s airborne virus from the respiratory tract, prevents fatalities for 10 years or more.
smallpox virus introduced into Central and skin lesions, and contaminated bedding or Smallpox is prevented even when the
South America caused horrendous loss of life other objects. vaccine is administered up to 4 days after
and may have contributed to the downfall of ■ It can be highly lethal, with mortality exposure to the virus.
the Inca and Aztec nations. An epidemic that rates generally above 25%. After the virus ■ Infected people do not spread the
swept the Massachusetts coast in the 1600s establishes infection of the respiratory disease during the long incubation
killed so many Native Americans that in some system, it enters the lymphatics and period, generally 12 to 14 days; they
communities there were not enough survivors bloodstream, finally causing lesions of the become infectious only with the onset
to bury the dead. Even in the latter 1700s, skin and throughout the body. of fever.
during the Revolutionary War, a smallpox epi- ■ The virus is relatively stable, probably ■ The disease can usually be diagnosed
demic raged through the American colonies. remaining infective for hours in the air rapidly, by the characteristic appearance
General George Washington suspected that of a building; viable smallpox has been of skin lesions that predominate on
the virus had been deliberately introduced by demonstrated in dried crusts from skin the face and hands, and by laboratory
the British. So many of his men were ill after lesions after storage for 10 years at room examination of material from skin
his defeat at Quebec in 1777 that he ordered temperature in ordinary envelopes. lesions.
the mass variolation of remaining troops.
■ Large numbers of people are highly ■ There is already widespread experience
smallpox, pp. 419, 447 variolation, p. 419
susceptible to the virus. Routine on how to watch for and contain the
Why does the ghost of smallpox concern
vaccination against smallpox was disease.
us now, when the last case, acquired through
discontinued in the United States several In a simulated attack on an American city in
a laboratory accident, occurred in 1978? The
decades ago. the summer of 2001, only 24 primary cases of
answer is that the smallpox virus still exists,
■ The relatively large genome of the smallpox increased in 2 months to 3 million,
locked in high-security laboratories in the
United States and the Russian Federation, and smallpox virus probably permits genetic with 1 million deaths. This study probably
perhaps held in secret locations by countries modifications that could enhance its greatly exaggerated the risk. Nevertheless,
or individuals that could use it to harm others. virulence. even though unlikely, the potential danger
A number of factors are notable regarding the from a smallpox introduction has caused the
Factors That Discourage United States to begin preparations for this
possibility of the virus being used as an agent
Its Use possibility, including markedly expanding its
of bioterror.
■ Propagating the smallpox virus is stockpile of smallpox vaccine. More informa-
Factors That Might dangerous and requires advanced tion on smallpox is available at the text web-
Encourage Its Use knowledge and laboratory facilities. site (www.mhhe.com/nester7).
■ It spreads easily from person to person, ■ A proven highly effective vaccine
mainly through close contact with (vaccinia virus) is available. A protective

Pathogenesis infectious viral particles are produced in the nuclei of the nerve
The VZV enters the body through the respiratory tract, establishes cells. The virus particles are then carried to the skin by the normal
an infection, replicates, and travels to the skin via the bloodstream. circulation of cytoplasm within the nerve cell. With the appear-
After the dermis and epidermis are infected, the virus spreads to ance of the skin lesions, a prompt, intense secondary (memory)
adjacent cells, and the characteristic skin lesions appear. response of adaptive immunity begins. As immune cells accumu-
Stained preparations of infected cells show intranuclear late in the ganglion, the signs and symptoms of shingles quickly
inclusion bodies (pink bodies in the nucleus) where the virus disappear. Sometimes, however, the inflammatory response leads
reproduces. Some infected cells fuse together, forming multi- to scars and chronic pain. secondary response, p. 367 cell-mediated
nucleated giant cells. The infected cells swell and eventually lyse. immunity, p. 356

When an area of skin infection involves a sensory nerve ending,

the virus enters the nerve. From there, it travels to ganglia (singu- Epidemiology
lar: ganglion), which are small bulges in sensory nerves located Chickenpox is a notifiable disease, requiring that confirmed cases
near the spine. Conditions inside the ganglia prevent full expres- are reported to the CDC. However, many cases are so mild that
sion of the VZV genome. However, viral DNA is present and fully they go unreported. In the early 1990s, before a vaccine was avail-
capable of coding for mature infectious virus. The mechanism of able, there were approximately 4 million cases of chickenpox each
suppression of viral replication within the nerve cell is not known year in the United States, resulting in 10,000 hospitalizations and
but is probably under the control of immune cells. approximately 100 to 150 deaths. A decade after the release of the
Shingles is most likely to occur when cell-mediated vaccine, chickenpox caused fewer than half a million cases and 8
immunity declines. With a weakened cellular immune system, deaths each year.
 Part IV Infectious Diseases 537

VZV can be transmitted by both respiratory secretions and By preventing chickenpox, the vaccine reduces the chance
skin lesions. As with many diseases transmitted by the respiratory of developing shingles. Another vaccine is available to prevent
route, most cases occur in the winter and spring months. Humans shingles in individuals 60 years of age or older. This vaccine is
are the only reservoir, and the disease is highly contagious. The composed of the same attenuated virus used in the chickenpox
incubation period averages about 2 weeks, with a range of 10 to vaccine, but in a much higher dose. It is given in a single dose and
21 days. Individuals are infective from 1 to 2 days before the rash reduces the risk of developing shingles by 50%.
appears until all the lesions have crusted (usually 4 days after the People with impaired immunity are at risk of severe dis-
onset). reservoir, p. 438 seminated VZV infections. These individuals include newborns
The ability of the chickenpox virus to form a latent infection and people with cancer, AIDS, or organ transplants. Immuno-
allows it to persist within a population. By contrast, a virus such compromised individuals can be partially protected from severe
as measles spreads quickly through a population and infected indi- disease by being passively immunized with VZIG, a hyperim-
viduals become immune or die. When this happens, the measles mune globulin with high concentrations of antibody to VZV.
virus eventually runs out of available hosts and disappears from The main features of chickenpox are summarized in table  22.9.
the community. The chickenpox virus can reappear from cases of passive immunity, p. 420 hyperimmune globulin, p. 420
shingles if enough susceptible individuals are present. Shingles
occurs in about one out of three individuals in their lifetime, and
as many as 25% of these patients develop eye involvement. Rubeola (Measles)
Measles, “hard measles,” and “red measles” are common names
Treatment and Prevention for rubeola. One of the great success stories of the last half of the
The antiviral medications acyclovir and famciclovir, among oth- twentieth century was the dramatic reduction in measles cases
ers, are helpful in preventing and treating VZV infections. as a result of immunizing children with an attenuated vaccine
A safe attenuated chickenpox vaccine has been used in the against the disease. The number of cases has increased in recent
United States since 1995. All healthy children and adults without years, due in large part to some parents’ refusal to vaccinate their
a history of chickenpox are advised to receive the vaccine. HIV- children.
infected children and adults should receive the vaccine if their
immune system is still intact. The vaccine should not be given to Signs and Symptoms
people with immunodeficiencies although healthy, non-immune Measles begins with fever, runny nose, cough, and swollen,
contacts of such people should be vaccinated. attenuated red, weepy eyes. Within a few days, a fine red rash appears
vaccines, p. 423 on the forehead and spreads outward over the rest of the body

TABLE 22.9 Varicella (Chickenpox)

1 Varicella-zoster virus is inhaled; Signs and Itchy bumps and blisters in various
infects nose and throat. symptoms stages of development, fever;
latent infections can reactivate,
2 The virus infects nearby lymph
resulting in shingles (herpes
nodes, reproduces, and enters
1 zoster), years later.
the bloodstream.
3 Infection of other body cells 7 Incubation period 10 to 21 days
occurs, resulting in showers of Causative agent Varicella-zoster virus; enveloped
virions into the bloodstream. 6 5 double-stranded DNA virus of the
4 These virions cause successive herpesvirus family.
skin lesions, which evolve into 2
3 Pathogenesis Multiplication in the upper
blisters and crusts. respiratory tract followed by
5 Immune system eliminates dissemination via bloodstream to
the infection except for some the skin; cytopathic effect of virus
virions inside the nerve cells. 4 includes giant cell formation.

6 If immunity decreases with Epidemiology Highly infectious. Acquired by the

age or other reason, the virus respiratory route; people infected
persisting in the nerve ganglia with either chickenpox or shingles
can infect the skin, causing the only source; spread via skin
shingles (herpes zoster). lesions and respiratory secretions.
7 Transmission to others occurs Treatment Acyclovir or similar antiviral
from respiratory secretions and prevention medication for prevention and
and skin. treatment. Attenuated vaccine.
Passive immunization with zoster
immune globulin (VZIG) for
immunocompromised individuals.
538 Chapter 22 Skin Infections

FIGURE 22.18 A Pathogenesis

Child with Measles
(Rubeola) The rash is
Rubeola virus is acquired by the respiratory route. It replicates in the
usually accompanied by upper respiratory epithelium, spreads to lymphatic tissues, and even-
fever, runny nose, and a tually spreads to all parts of the body. The rash is caused by a cell-
bad cough. mediated immune response against viruses multiplying in the skin.
? Why is it important to Mucous membrane involvement is responsible for an impor-
continue immunizing tant diagnostic sign, Koplik spots (figure 22.19). These look like
children in the grains of salt lying on red and rough oral mucosa, resembling red
United States against
sandpaper, and are usually best seen in the back of the mouth,
measles, even though
it is a rare disease opposite the molars.
here? The measles virus temporarily suppresses cell-mediated
immunity, causing cold sores to appear and latent tuberculosis dis-
ease to reactivate in some individuals. It also damages the respi-
ratory mucous membranes, thereby increasing the susceptibility
of measles patients to secondary bacterial infections, especially
infection of the middle ear and lung. Damage to the intestinal
epithelium may explain the diarrhea that sometimes occurs in
measles and contributes to high measles death rates in impover-
ished countries. In the United States, deaths from measles occur
in about one to two of every 1,000 cases, mainly from pneumonia
(figure  22.18). Unless complications occur, signs and symp-
and encephalitis. cold sores, p. 581 tuberculosis, p. 502
toms generally disappear in about a week. Unfortunately, many
cases are complicated by secondary infections caused mainly by
Epidemiology
Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus
pyogenes, and Haemophilus influenzae. These bacteria easily Humans are the only natural host of rubeola virus. Before vaccina-
invade the body because the measles virus damages the normal tion became widespread in the 1960s, probably less than 1% of the
body defenses. Secondary infections most commonly cause ear- global population escaped infection with this highly contagious
aches and bacterial pneumonia. virus. Continued use of the measles vaccine resulted in a progres-
In about 5% of cases, the measles virus causes viral pneu- sive decline in the number of cases, so that measles is no longer
monia, with rapid breathing, shortness of breath, and dusky skin endemic in the Western Hemisphere. Small outbreaks can occur,
color from lack of adequate O2 exchange in the lungs. Encephalitis however, when the virus enters the area with people travelling
(inflammatory disease of the brain) is another serious complica- to and from other countries. In these situations, the virus spreads
tion, marked by fever, headache, confusion, and seizures. This among non-immune populations including (1) children too young
complication occurs in about one out of every 1,000 measles to be vaccinated; (2) children and adults inadequately vaccinated;
cases, and sometimes results in permanent brain damage, with and (3) unvaccinated people.
mental disability, deafness, and epilepsy. Worldwide, about 750,000 children still die from measles
Very rarely, measles is followed 2 to 10 years later by a every year. This ranks among the leading causes of death and dis-
disease called subacute sclerosing panencephalitis (SSPE). This ability among the impoverished. The case-fatality rate may reach
disease is marked by slowly progressive degeneration of the brain, 15% and the rate of secondary infections may reach 85% when
generally resulting in death within 2 years. Defective measles virus patients are poorly nourished. Measles vaccination is a high prior-
particles that cannot complete replication can be detected in the ity in areas struck by natural disasters.
brains of these patients. High levels of measles antibodies are pres-
ent in their blood. Successful immunization programs in the United
States have decreased SSPE, but there is concern that the recent
increase in measles cases may lead to more SSPE in the future.
Measles that occurs during pregnancy increases the risk of Koplik spots
miscarriage, premature labor, and low birth weight babies. Birth
defects, however, generally do not occur.

Causative Agent
Measles is caused by rubeola virus, an enveloped, single-stranded
RNA virus of the paramyxovirus family. Two biologically impor-
tant proteins are found on the viral envelope—hemagglutinin and
fusion protein. The virus uses hemagglutinin to attach to host
cells and the fusion protein to fuse the viral envelope with the
host cell’s cytoplasmic membrane. The fusion protein also causes FIGURE 22.19 Koplik Spots, Characteristic of Measles
adjacent infected host cells to join together, producing multinucle- (Rubeola) These spots resemble grains of salt on a red base.
ated giant cells. ? Can someone have measles without having Koplik spots?
 Part IV Infectious Diseases 539

TABLE 22.10 Rubeola (Measles)

1 Airborne rubeola virus infects

5 Signs and Rash, fever, weepy eyes, cough,
eyes and upper respiratory 4 symptoms and nasal discharge
tract, then the lymph nodes in Incubation period 10 to 12 days
the region. 1
Causative agent Rubeola virus, a single-stranded
2 Virus enters the bloodstream
7 RNA virus of the paramyxovirus
and is carried to all parts of
family
the body including the brain,
lungs, and skin. 4 Pathogenesis Virus multiplies in respiratory
tract; spreads to lymphatic tissues,
3 Skin cells infected with the
6 then to all parts of body, notably
rubeola virus are attacked 2
skin, lungs, and brain; damage to
by cytotoxic T cells, causing a
respiratory tract epithelium leads
generalized rash.
to secondary infection of ears and
4 Virus replicating in the lungs 3 lungs.
can cause pneumonia; the
Epidemiology Acquired by respiratory route;
brain can also be infected.
highly contagious; humans only
5 In rare cases, virus persisting source.
in the brain causes subacute
Treatment No antiviral treatment currently
sclerosing panencephalitis,
and prevention available. Attenuated vaccine
months or years after the acute
after age 12 months; second dose
infection.
on entering elementary school or
6 Secondary infection of the ears at adolescence.
and lungs is common.
7 Transmission is by respiratory
secretions.

Treatment and Prevention Signs and Symptoms

No antiviral treatment currently exists for measles, but an attenu- Characteristic signs and symptoms of German measles are slight
ated vaccine can prevent the disease. In 1980, the worldwide fever, mild cold symptoms, and enlarged lymph nodes behind the
incidence of measles was estimated to be 100 million with 5.8 mil- ears and on the back of the neck. After about a day, a faint rash
lion deaths, but vaccination programs have lowered the number consisting of many pink spots appears over the face, chest, and
of cases dramatically since then. Unfortunately, the incidence of abdomen (figure 22.20). Unlike measles, there are no diagnostic
measles in the United States is on the rise. This is largely caused mouth lesions. Adults commonly develop painful joints, with pain
by importation and spread of the disease by travelers from coun- generally lasting 3 weeks or less. Other signs and symptoms typi-
tries where it is still endemic. cally last only a few days. The significance of German measles,
The measles vaccine is usually given together with mumps, however, lies with its threat to a developing fetus.
rubella, and varicella vaccines (MMRV). The first injection of
vaccine is given near an infant’s first birthday. Since 1989, a sec-
ond injection of vaccine is given when children enter elementary
school. The two-dose regimen has resulted in at least 99% of
the recipients becoming immune. In an epidemic, vaccine is given
to babies as young as 6 months, who are then reimmunized before
their second birthday. Students entering high school or college
are often required to get a second dose of vaccine if they have not
had one earlier. Those at high risk of acquiring measles, such as
medical personnel, should be immunized unless they definitely
have had the disease or have laboratory proof of immunity. Some
features of measles are summarized in table 22.10.

Rubella (German Measles)

German measles and “three-day measles” are common names for
rubella. The term German measles arose because the disease was FIGURE 22.20 Adult with German Measles (Rubella)
first described in Germany. In contrast to chickenpox and measles, Symptoms are often very mild, but the effects on a fetus can be
rubella is typically a mild, often unrecognized disease that is dif- devastating.
ficult to diagnose. It is of concern because infection of pregnant ? Is congenital rubella syndrome more likely to occur if a pregnant
women can have tragic consequences. woman is infected early or late in the pregnancy?
540 Chapter 22 Skin Infections

CASE PRESENTATION
The patient was a 20-year-old asymptomatic showed it to be the vaccine strain of mea- vaccine virus, is declining, but outbreaks
man who was immunized against measles sles virus. The patient received intravenous in colleges and other institutions still
as a requirement for starting college. He had immune globulin and an antiviral medica- occur. A severely immunodeficient indi-
received his first dose of measles vaccine at tion—ribavirin—and improved. Subsequently, vidual can be passively immunized against
approximately 1 year of age. Past medical his- however, his condition deteriorated, and he measles with immune globulin if exposed
tory revealed that he was a hemophiliac and died of presumed complications of AIDS. to the wild-type virus.
had contracted the human immunodeficiency 1. Is measles immunization a good idea for 2. Measles is often given as an example of
virus (HIV) from clotting factor (a blood prod- people with immunodeficiency? a persistent viral infection, meaning that
uct given to control bleeding) contaminated following infection the virus can persist in
2. Is it surprising that the vaccine virus was
with the virus. Laboratory tests showed that he the body for months or years in a slowly
still present in this patient 11 months after
had a very low CD4+ lymphocyte count, indi- replicating form. It has been suggested but
vaccination? Explain.
cating a severely damaged immune system. not proven that this explains the lifelong
About a month after his precollege immu- 3. Despite the severe infection, there was no
immunity conferred by measles infection
nization, he developed Pneumocystis pneumo- rash. Why?
in normal people. Rarely in presumably
nia, a lung infection characteristic of AIDS, normal individuals and, more commonly,
was hospitalized, had a good response to treat- Discussion in malnourished or immunodeficient indi-
ment, and was discharged. Ten months later, 1. Measles is often disastrous for persons viduals, persistent infection leads to dam-
he was again hospitalized for symptoms of a with AIDS or other immunodeficiencies. age to the brain, lung, liver, and possibly,
severe lung infection. He had no rash. Multiple They should be immunized as soon as pos- the intestine.
laboratory tests to determine the cause of sible in their illness, before the immune 3. Following acute infection, the measles
his infection were negative. Finally, a lung system becomes so weakened it cannot virus floods the bloodstream and is carried
biopsy was performed and revealed “giant respond effectively to the vaccine. Also, to various tissues of the body, including
cells”—very large cells with multiple nuclei. as this and other cases have shown, the the skin. The rash that characterizes mea-
Cytoplasmic and intranuclear inclusion bod- vaccine virus can itself cause disease when sles is caused by T cells attacking measles
ies were also present. This picture was highly immunodeficiency is severe. With the virus antigen lodged in the skin capillaries.
suggestive of measles pneumonia, and measles worldwide effort to eliminate measles, the In the absence of functional T cells, the
virus subsequently was recovered from cell risk of exposure to the wild-type measles rash does not occur.
cultures of the biopsy material. Other studies virus, as opposed to the laboratory-derived

Causative Agent that live continue to excrete rubella virus in throat secretions and
German measles is caused by the rubella virus, a member of the urine for many months. The likelihood of the syndrome varies
togavirus family. The virus is an enveloped, single-stranded RNA according to the age of the fetus when infection occurs. Infections
virus that can easily be cultivated in cell cultures. Proteins on the within the first 6 weeks of pregnancy result in most of the fetuses
viral surface cause in vitro hemagglutination, which can be inhib- having a detectable injury, commonly deafness. Even infants who
ited by specific antibody. This allows serological identification of are apparently normal, however, excrete rubella virus for extended
the virus. periods and thus can infect others.
Epidemiology
Pathogenesis
Humans are the only natural host for rubella virus. The disease is
The rubella virus enters the body via the respiratory tract. It highly contagious although less so than measles; it is estimated that
multiplies in the nasopharynx and enters the bloodstream, caus- in the prevaccine era, only 10% to 15% of people reached adulthood
ing a sustained viremia. The virus travels to various body tis- without being infected. Complicating the epidemiology of rubella is
sues, including the skin and joints. Humoral and cell-mediated the fact that over 40% of infected individuals fail to develop symp-
immunity develop against the virus, and the resulting immune toms, but can still spread the virus. People who develop typical
complexes probably cause the rash and joint symptoms. viremia, rubella can be infectious for as long as 7 days before the rash appears
p. 384 immune complexes, p. 395
until 7 days afterward. Before widespread use of the vaccine began
If a woman is infected early in the pregnancy, virus particles in 1969, periodic major epidemics arose. One epidemic in 1964
in the bloodstream can cross the placenta and infect the fetus. resulted in about 30,000 cases of congenital rubella syndrome. This
This is less likely to happen later in pregnancy. Virtually all types caused a “rubella bulge” of hearing-impaired children that affected
of fetal cells are susceptible to infection; some cells are killed, the educational system in the United States for decades.
whereas others develop a persistent infection in which cell division
is impaired and chromosomes are damaged. The result is a charac- Treatment and Prevention
teristic pattern of fetal abnormalities referred to as the congenital No specific antiviral therapy for rubella is available. The disease
rubella syndrome. The abnormalities include cataracts and other can be prevented by an attenuated rubella virus given to babies at
eye defects, brain damage, deafness, heart defects, and low birth 12 to 16 months old with a second dose at age 4 to 6 years. The
weight despite normal gestation. Babies may be stillborn. Those vaccine produces long-lasting immunity in nearly all recipients.
 Part IV Infectious Diseases 541

FIGURE 22.21 Reported Cases

100,000 80 of German Measles (Rubella)
and Congenital Rubella
Syndrome (CRS), United States,
1966–2009
10,000
Number of rubella cases

60

Number of CRS cases

? Why are there fewer cases of
Rubella CRS than of German measles at
1,000 every time tested?

40
100

CRS 20
10

0 0
1966 1970 1974 1978 1982 1986 1990 1994 1998 2002 2006
Year

The vaccine is not given to pregnant women for fear it might result about 50 members associated with skin lesions. In the early
in congenital defects. As an added precaution, women are advised 1900s the causes of the common childhood rashes were largely
not to become pregnant for 28 days after receiving the vaccine. unknown, and it was the practice to number them 1 to 6 as follows:
Use of the vaccine has markedly reduced the incidence of (1) rubeola; (2) scarlet fever; (3) rubella; (4) Duke’s disease—a mild
rubella in the United States to generally less than 50 cases per year disease with fever and bright red generalized rash, now thought to
(figure 22.21). There were less than 1,000 confirmed cases in the have been due to an enterovirus; (5) erythema infectiosum; and
entire Western Hemisphere in 2004. Some features of German (6) exanthem subitum. The causes of erythema infectiosum (fifth
measles are summarized in table 22.11. disease) and exanthem subitum (roseola) have only been estab-
lished in recent years.
Fifth disease (erythema infectiosum) occurs in both chil-
Other Viral Rashes of Childhood dren and young adults. The illness begins with fever, malaise,
The kinds of viruses that can cause childhood rashes probably and head and muscle aches. A diffuse redness appears on the
number in the hundreds. One group alone, the enteroviruses, has cheeks, giving the appearance of the face as if it were slapped

TABLE 22.11 Rubella (German Measles)

1 Airborne rubella virus infects Signs and symptoms Mild fever and cold symptoms,
nose and throat. rash beginning on forehead
2 Virus taken up by lymph nodes and face, enlarged lymph nodes
in the region. behind the ears
1 Incubation period 14 to 21 days
3 Rubella virus multiples and
7
enters the bloodstream. 2 Causative agent Rubella virus, an RNA virus of the
4 Circulating virus reacts with togavirus family
antibodies, resulting in Pathogenesis Following replication in the upper
immune complexes. respiratory tract, virus spreads to
5 Immune complexes lodge in 3 all parts of the body and crosses
4 the placenta; surviving fetuses
the skin, causing a rash, and
in the joints, causing pain. often develop abnormally, and
they excrete the virus for months
6 In pregnant women, rubella after birth.
virus crosses the placenta, 5
6 Epidemiology Virus possibly present in nose and
infecting the fetus, resulting in
congenital rubella syndrome. throat from 1 week before rash to
1 week after; infection occurs via
7 Transmission to others is by the respiratory route; humans are
respiratory secretions. the only source.
Treatment and No specific antiviral treatment.
prevention Attenuated vaccine administered
to children at 12 to 16 months,
repeated at 4 to 6 years of age.
542 Chapter 22 Skin Infections

(a) (b)

FIGURE 22.22 Erythema Infectiosum (Fifth Disease) (a) “Slapped cheek” appearance of the rash on the face. (b) Appearance of the rash
on the extremities.
? Why is this disease a major threat to individuals with sickle cell anemia?

(figure  22.22a). The rash commonly spreads in a lacy pattern to Papillomaviruses can survive on inanimate objects such as wres-
involve other parts of the body, especially the extremities (figure  tling mats, towels, and shower floors, and infection can be acquired
22.22b). The rash may come and go for 2 weeks or more before from such contaminated objects. The viruses infect the deeper cells
recovery. Adults with fifth disease often have joint pains. The dis- of the epidermis and reproduce in the nuclei. Some of the infected
ease is caused by parvovirus B-19, a naked, single-stranded DNA cells grow abnormally, forming the wart. The incubation period
virus. The virus preferentially infects certain bone marrow cells and ranges from 2 to 18 months. Infectious virus is present in the wart
is a major threat to people with sickle cell and other anemias. The and can contaminate fingers or objects that pick or rub the lesions.
infected marrow sometimes stops producing blood cells, a condi- Like other tumors, warts can be treated effectively only by kill-
tion known as aplastic crisis. Also, about 10% of women infected ing or removing all of the abnormal cells. This can usually be done
with the virus during pregnancy suffer spontaneous abortion. by freezing the wart with liquid nitrogen, by cauterization (burning
Roseola (exanthem subitum, roseola infantum), is a common the tissue usually with an electrically heated needle), or by surgically
disease in children 6 months to 3 years old. It causes a great deal
of parental anxiety because it begins abruptly with a fever that
may reach 105°F and cause convulsions. The children generally
do not appear ill, however. After several days, the fever goes
away and a short-lived red rash appears, mainly on the chest and
abdomen. The patient has no additional signs or symptoms at this
point, and the rash vanishes in a few hours to 2 days. This disease
is caused by herpesvirus type 6. There is no vaccine against the
disease, and no treatment except to reduce the risk of seizures by
sponging with lukewarm water and using medication to keep the
temperature below 102°F.

Warts
Papillomaviruses cause warts by infecting the skin through minor
abrasions. Warts are small tumors called papillomas, and con-
sist of multiple protrusions of tissue covered by skin or mucous
membrane. Warts rarely become cancers, although some sexually
transmitted papillomaviruses cause cervical cancer. About half of
the time, warts on the skin disappear within 2 years without any
treatment. Papillomaviruses (figure  22.23) belong to the papo-
vavirus family. They are naked, double-stranded DNA viruses. 1 μm

More than 50 different papillomaviruses infect humans, but they

are difficult to study because they grow poorly in cell cultures or FIGURE 22.23 Wart Virus The virions appear yellow in this color-
experimental animals. Warts of other animals are generally not enhanced transmission electron micrograph.
infectious for humans. cervical cancer, p. 631 ? Which group of viruses cause warts?
 Part IV Infectious Diseases 543

removing the wart. Virus generally remains in the adjacent normal- and vesicles develops distant from the infected area. This rash is
appearing skin and may cause additional warts. referred to as a dermatophytid, or “id” reaction, a reflection of
Warts that grow on the soles of the feet are called plantar allergy to products of the infecting fungus.
warts (often mistakenly called planter’s warts; plantar is a word
meaning “referring to the sole of the foot”). These warts are very Causative Agents
difficult to treat because the pressure of standing on them causes Dermatophytes are a group of skin-invading molds including
them to grow wide and deep. members of the genera Epidermophyton, Microsporum, and
Trichophyton (figure  22.24). They can be grown on media
MicroAssessment 22.3 especially designed for molds and are usually identified by
Chickenpox, measles, and rubella can be controlled by vaccines. their colonial and microscopic appearance. In some cases their
Viral diseases that may only inconvenience a pregnant woman can nutritional requirements and biochemical tests are important
be disastrous to her fetus. A viral infection acquired in childhood as well.
can remain latent for years only to reactivate in a different form. One
group of viruses causes benign skin tumors. Pathogenesis
7. What important diagnostic sign is often present in the mouth of The normal skin is generally resistant to invasion by dermato-
measles (rubeola) patients? phytes. Some species, however, are relatively virulent and can
8. Does exposure to a person with shingles transmit shingles or even cause epidemics, especially among children. In moist areas
chickenpox? of the skin, dermatophytes can invade keratin-containing cells
9. Why is it a good idea to immunize both boys and girls and structures. They produce an enzyme called keratinase that
against rubella? +

22.4 ■ Skin Diseases

Caused by Fungi
Learning Outcomes
9. Describe the characteristics of superficial cutaneous mycoses,
including the role of dermatophytes.
10. Describe the condition tinea versicolor and its causative agent,
Malassezia furfur.

Diseases caused by fungi are called mycoses. Several fungi are

responsible for mild to serious infections of the skin. The condi-
tion of the host’s defenses against infection determines the sever-
ity of most fungal infections.
(a)

Superficial Cutaneous Mycoses

A group of molds called dermatophytes can invade hair, nails,
and the keratinized portion of the skin. The resulting mycoses
have colorful names such as jock itch, athlete’s foot, and ring-
worm, as well as Latinized names that describe their location:
tinea capitis (scalp), tinea barbae (beard), tinea axillaris (armpit),
tinea corporis (body), tinea cruris (groin), and tinea pedis (feet), to
list a few. Tinea simply means “worm,” which probably reflects
early incorrect ideas about the cause.

Signs and Symptoms

Most people colonized by dermatophytes have no signs or symp-
toms at all. Others complain of itching, a bad odor, or a rash. In
ringworm, a rash occurs at the site of the infection and consists of
(b) 20 μm
a scaly area surrounded by redness at the outer margin, producing
irregular rings or a lacy pattern on the skin. On the scalp, patchy FIGURE 22.24 Dermatophytosis (a) Tinea pedis, usually
areas of hair loss can occur, with a fine stubble of short hair left caused by species of Trichophyton. (b) Large boat-shaped conidia of
behind. Infected nails become thickened and brittle and may sepa- Microsporum gypseum, a cause of scalp ringworm in children.
rate from the nailbed. Sometimes, a rash consisting of fine papules ? What is the common name for tinea pedis?
544 Chapter 22 Skin Infections

FIGURE 22.25 Tinea

Versicolor Appearance in
(a) a fair-skin individual
and (b) a dark-skin individual.
(c) Microscopic appearance
of skin scraping showing
Malassezia furfur yeast and
filamentous forms.
? What other conditions
can be caused by this
fungus?

(a) (b)

breaks down the protein, allow- Treatment and Prevention

ing them to use it as a nutrient. Numerous prescription and over-the-counter medications can be
Dermatophytes can invade the used to treat superficial skin infections. However, nail infections
epidermis down to the level are often much more difficult to cure, requiring taking medication
of the keratin-producing cells. by mouth for months, and sometimes surgical removal of the nail.
Hair is invaded at the follicle, Attention to cleanliness and maintenance of normal dry-
which is relatively moist. ness of the skin and nails effectively prevent most dermatophyte
Fungal products diffuse into infections. Powders, open shoes, changing of socks, and applying
the dermis and provoke an immune rubbing alcohol after bathing may help prevent toenail infections.
(c) reaction, which probably explains
why adults tend to be more resistant to
Other Fungal Diseases
infection than children. Children are more likely
to have hypersensitivities, like asthma and eczema. Diffusion of Although Malassezia furfur is generally harmless and commonly
fungal products also explains why some people develop the allergic found on the skin, in some people, it causes skin conditions such
“id” reactions. as a scaly face rash, dandruff, or tinea versicolor (figure 22.25).
The latter is a common skin disease characterized by patchy scali-
Epidemiology ness and increased pigment in light-skin people, or a decrease in
Patient age, virulence of the infecting strain of mold, and moisture pigment in dark-skin people. Scrapings of the affected skin show
availability are important factors in determining the course of large numbers of M. furfur, both in its yeast form and as short fila-
infection. Common causes of excessive moisture include obesity ments called hyphae. Unknown host factors are important in these
where folds of skin lie together, tight clothing, and plastic or rub- diseases because most people carry the organism on their skin
ber footwear. Potentially pathogenic molds may be present in soil without any disease. AIDS patients often have a severe rash with
and on pets such as young cats and dogs. Fungi acquired through pus-filled pimples caused by Malassezia yeasts, and the organ-
soil or animal contact tend to cause more noticeable signs and isms may even infect internal organs in patients receiving lipid-
symptoms in humans. containing intravenous feedings. yeasts, p. 283 hyphae, p. 284

(a) (b) 10 mm
FIGURE 22.26 Candida albicans (a) Causing a diaper rash; (b) Gram stain of pus showing C. albicans yeast forms and filamentous forms called
pseudohyphae.
? Would an antibiotic ointment containing penicillin be effective against this candidal infection?
Diseases in Review 22.1
Common Bacterial, Viral, and Fungal Skin Diseases

Disease Causative Agent Comment Summary Table

BACTERIAL SKIN DISEASES

Acne Propionibacterium acnes Most common during puberty, probably due to excess sebum
commonly associated secretion in response to increased hormone levels.
Hair follicle infections Staphylococcus aureus Causative agent commonly colonizes the nostrils and moist skin Table 22.2, p. 524
areas; skin infections include folliculitis, furuncles, and carbuncles.
Organism is often resistant to multiple antibiotics.
Staphylococcal scalded Exfoliatin-producing Characterized by peeling of the outer layer of skin; Table 22.4, p. 528
skin syndrome strains of S. aureus occurs in newborns and infants, as well as the elderly and
immunocompromised.
Impetigo Usually Streptococcus Characterized by blisters, which break and are replaced by oozing Table 22.6, p. 529
pyogenes; sometimes yellow crusts; some people later develop glomerulonephritis.
Staphylococcus aureus
Rocky Mountain spotted Rickettsia rickettsii Spread by ticks; characterized by rash that spreads and then Table 22.7, p. 531
fever (RMSF) becomes hemorrhagic.
Lyme disease Borrelia burgdorferi Spread by ticks; characterized by “bull’s-eye rash”; later symptoms Table 22.8, p. 534
include injury to heart and nervous system, and arthritis.

VIRAL SKIN DISEASES

Varicella Varicella-zoster virus (VZV) Virus enters via the respiratory tract; infection characterized by Table 22.9, p. 537
(chickenpox) itchy skin lesions; VZV becomes latent and can later reactivate to
cause shingles. Preventable by vaccination.
Rubeola (measles) Rubeola virus Virus enters via the respiratory tract. Disease manifests with Table 22.10, p. 539
respiratory symptoms and a spreading rash; Koplik spots occur in
mouth. Preventable by vaccination.
Rubella (German measles) Rubella virus Virus enters via the respiratory tract, causes mild respiratory Table 22.11, p. 541
symptoms, joint pain, and fine rash; can damage developing fetus
(congenital rubella syndrome). Preventable by vaccination.
Warts (dermal warts) Papillomaviruses Warts, which are benign skin tumors, can be removed by
freezing, burning, surgery, or topical medication.

FUNGAL SKIN DISEASES

Superficial cutaneous Usually Epidermophyton, Fungi invade keratinized skin, causing what is commonly known
mycoses Microsporum, or as athlete’s foot, jock itch, and ringworm.
Trichophyton species
Other fungal diseases Malassezia furfur, Candida Both organisms are usually harmless on the skin but M. furfur
albicans sometimes causes skin conditions such as scaly face rash,
dandruff, or tinea versicolor. C. albicans sometimes invades
deeper layers and subcutaneous tissues.

The yeast Candida albicans may live harmlessly among the MicroAssessment 22.4
normal microbiota of the skin, but in some people it invades the
deep layers of the skin and subcutaneous tissues (figure  22.26). The fungi that cause skin mycoses can commonly colonize skin
without causing signs or symptoms. The best protection against
In many people with candidal skin infections, no precise cause for fungal skin infections is to maintain normal skin dryness.
the invasion can be determined. Certain molds also cause cutane-
10. What is a mycosis?
ous mycoses, but they are not as likely as C. albicans to invade
the deep skin layers. 11. What kinds of structures are invaded by dermatophytes?
12. Why do you think it is so more difficult to treat nail infections
The key features of the diseases covered in this chapter are high- than other superficial dermatophytoses? +
lighted in the Diseases in Review 22.1 table.
546 Chapter 22 Skin Infections

FUTURE CHALLENGES 22.1

The Ecology of Lyme Disease
Lyme disease is often referred to as one corresponding increase in Ixodes scapularis burgdorferi bacteremia following infection
of the emerging diseases. Unrecognized in ticks. Both deer and mice feed on the acorns from a tick, and the blood of a common liz-
the United States before 1975, it is now and subsequently spread the disease to adja- ard host along the West Coast even kills the
the most commonly reported vector-borne cent areas. Variations in weather conditions, spirochetes. The role of snakes, foxes, and
disease. Because of the seeming explosion and their effect on food supply for these ani- birds of prey that control mouse populations
in the numbers of Lyme disease cases, and mals, might therefore be an important ecologi- and that of birds, spiders, and wasps that feed
its apparent extension to new geographical cal factor, although it is not clear that weather on ticks are also under study. The challenge
areas, the ecology of Lyme disease is under cycles completely explain the emerging nature is to define more completely the ecology of
intense study. In the northeastern United of the disease. The presence of animals other Lyme and other tick-borne diseases in order
States, large increases in white-footed mouse than white-footed mice for the ticks to feed to predict their emergence and find new ways
populations occur in oak forests during years on is another factor. Alternative tick hosts for their prevention.
in which there is a heavy acorn crop, with a usually do not have a sustained Borrelia

Summary
22.1 ■ Anatomy, Physiology, and Ecology 22.3 ■ Skin Diseases Caused by Viruses
The skin prevents the entry of microbes, regulates body temperature,
Varicella (Chickenpox) (table 22.9)
restricts the loss of fluid from body tissues, and plays an essential role
in the function of the immune system. It is composed of the epidermis Chickenpox, once a common disease of childhood, is caused by the
and the dermis (figure 22.1). Common members of the skin microbiota varicella-zoster virus (figure 22.15). Shingles can occur months or years
include diphtheroids, staphylococci, and fungi (table 22.1). after chickenpox and is due to reactivation of the virus (figure  22.16).
Shingles cases can be sources of chickenpox epidemics.

22.2 ■ Bacterial Skin Diseases Rubeola (Measles) (table 22.10)

Rubeola (measles) is a potentially dangerous viral disease that can lead
Acne Vulgaris
to serious secondary bacterial infections, and fatal lung or brain dam-
Acne is characterized by enlarged sebaceous glands. Sebum accumu- age. Measles can be controlled by vaccinating with an attenuated vac-
lation within these glands allows Propionibacterium acnes to grow cine (figures 22.18, 22.19).
to high numbers. Their metabolic products cause an inflammatory
response. Rubella (German Measles) (table 22.11)
German measles (rubella), if contracted by a woman early in preg-
Hair Follicle Infections nancy, often results in birth defects, making up the congenital rubella
Folliculitis, boils and carbuncles are caused by Staphylococcus aureus syndrome. Immunization with an attenuated virus protects against this
which is coagulase-positive and often resists penicillin and other anti- disease (figure 22.21).
biotics (figures 22.2 and 22.3, table 22.3). There are many different strains Other Viral Rashes of Childhood
that vary in virulence. A carbuncle is more serious because the infec- Numerous viruses can cause rashes. Fifth disease (erythema infectio-
tion is more likely to be carried to the heart, brain, or bones. sum), caused by parvovirus B-19, is characterized by a “slapped cheek”
rash (figure  22.22). Roseola (exanthem subitum), caused by
Staphylococcal Scalded Skin Syndrome (table 22.4)
herpesvirus type 6, is marked by a high fever and a rash that appears
Staphylococcal scalded skin syndrome results from exfoliatin produced as the temperature returns to normal.
by certain strains of Staphylococcus aureus (figure 22.4).
Warts
Streptococcal Impetigo (table 22.6) Warts are skin tumors caused by a number of papillomaviruses
Impetigo is a superficial skin infection caused by Streptococcus (figure 22.23). They rarely become cancers, but some sexually transmit-
pyogenes and Staphylococcus aureus (figure 22.5). ted papillomaviruses cause cervical cancer.

Rocky Mountain Spotted Fever (table 22.7; figure 22.6) 22.4 ■ Skin Diseases Caused by Fungi
Rocky Mountain spotted fever, caused by the obligate intracellular Superficial Cutaneous Mycoses
bacterium Rickettsia rickettsii, is an often fatal disease transmitted to Dermatophytes cause athlete’s foot, ringworm, and invasions of the
humans by the bite of an infected tick (figure 22.8). hair and nails (figure 22.24).
Lyme Disease (table 22.8) Other Fungal Diseases
Lyme disease is characterized by stages of disease and is caused by a Malassezia sp. can cause tinea versicolor and dandruff, as well as seri-
spirochete, Borrelia burgdorferi (figure  22.11). A bull’s-eye rash is the ous skin disease in AIDS patients (figure 22.25). Candida albicans may
hallmark of the early stage disease (figure 22.10). B. burgdorferi is trans- live harmlessly among the normal flora, but it can invade deeper layers
mitted by certain ticks (figures 22.13, 22.14). of the skin and subcutaneous tissues (figure 22.26).
 Part IV Infectious Diseases 547

Review Questions
Short Answer 7. All of the following are true of Lyme disease except
1. What is the difference between a furuncle and carbuncle? a) it is caused by a spirochete.
2. Why do only certain strains of Staphylococcus aureus cause b) it is transmitted by certain species of ticks.
scalded skin syndrome? c) it occurs only in the region around Lyme, Connecticut.
3. How is impetigo spread? d) most cases get a rash that looks like a target.
e) it can cause heart and nervous system damage.
4. How does the fact that Rocky Mountain spotted fever is a zoonosis
relate to the relative severity of the disease symptoms? 8. Which of the following statements is more likely to be true of
measles (rubeola) than German measles (rubella)?
5. Describe the causative agent of Lyme disease.
a) Koplik spots are present.
6. What is characteristic about the rash of varicella?
b) It causes birth defects.
7. What is the relationship between chickenpox (varicella) and
c) It causes only a mild illness.
shingles (herpes zoster)?
d) Human beings are the only natural host.
8. Why are many cases of measles complicated by secondary
e) Attenuated virus vaccine is available for prevention.
infections?
9. All of the following must be cultivated in cell cultures instead of
9. What is the significance of rubella viremia during pregnancy?
cell-free media except
10. How does a person contract warts?
a) Rickettsia rickettsii. b) rubella virus.
Multiple Choice c) varicella-zoster virus. d) Borrelia burgdorferi.
1. Which of the following conditions is important in the ecology of e) rubeola virus.
the skin? 10. All of the following might contribute to development of ringworm
a) Temperature b) Salt concentration c) Lipids or other superficial cutaneous mycoses except
d) pH e) All of the above a) obesity. b) playing with kittens.
2. Staphylococcus aureus can be responsible for which of these c) rubber boots. d) using skin powder.
following conditions? e) dermatophyte virulence.
a) Impetigo b) Food poisoning
Applications
c) Toxic shock syndrome d) Scalded skin syndrome
1. A school administrator in a small Iowa community prohibited a
e) All of the above
child with chickenpox from attending school. He said this was the
3. The main effect of staphylococcal protein A is to first case of chickenpox in the school in 6 years and he did not want
a) interfere with phagocytosis. to have an outbreak. Several parents argued to the school board
b) enhance the attachment of the Fc portion of antibody to that an outbreak would benefit the school in the long term. Discuss
phagocytes. the pros and cons of allowing this child to attend school.
c) coagulate plasma. 2. A public health official was asked to speak about immunization
d) kill white blood cells. during a civic group luncheon. One parent asked if rubella was
e) degrade collagen. still a problem. In answering the question, the official cautioned
women planning to have another child to have their present chil-
4. Which of the following is essential for the virulence of Streptococcus
dren immunized against rubella. Why did the official suggest this?
pyogenes?
a) Protease b) Hyaluronidase c) DNase Critical Thinking +
d) All of the above e) None of the above 1. When Lyme disease was first being investigated, the observation
5. Which of the following statements is true of streptococcal impetigo? that frequently only one person in a household was infected was a
a) It is caused by a Gram-negative rod. clue leading to the discovery that the disease was spread by arthro-
b) It cannot be transmitted from one person to another. pod bites. Why was this so?
c) Pathogenic streptococci all produce coagulase. 2. Why might it be more difficult to eliminate a disease like Lyme dis-
d) All of the above. ease or Rocky Mountain spotted fever from the earth than rubeola
e) None of the above. or rubella?
6. All of the following are true of Rocky Mountain spotted fever
except
a) the disease is most prevalent in the western United States.
b) it is caused by an obligate intracellular bacterium.
c) it is a zoonosis transmitted to human beings by ticks.
d) those with the disease characteristically develop a hemorrhagic
rash.
e) antibiotic therapy is usually curative if given early in the
disease.
 23 Wound Infections
KEY TERMS
Abscess A localized collection of
pus within a tissue.
Fasciitis Inflammation of the fascia,
which are bands of fibrous tissue
Pus Yellowish fluid composed of
proteins, living and dead leukocytes,
and tissue debris that accumulates
as a result of pyogenic bacterial
infections.
that underlie the skin and surround
muscle and body organs. When Pyogenic Pus-producing.
fasciitis leads to death of tissue, it is Superantigen Molecules that
called necrotizing fasciitis. bind to and stimulate helper T cells,
Granulation Tissue New tissue resulting in activation of large
formed during healing of an injury, numbers of T cells, overproduction
consisting of small, red, translucent of cytokines, and sometimes fatal
nodules containing many blood shock.
vessels. Synergistic Infection An
MRSA Methicillin-resistant infection in which two or more
Staphylococcus aureus; many species of pathogens act together to
strains have acquired R plasmids, produce an effect greater than the
making, them resistant to multiple sum of effects if each pathogen were
antimicrobial drugs. acting alone.

Shotgun wound of the torso.

where he worked and trained Japanese scientists for the rest of his life.
In 1908, Koch visited Kitasato in Japan and a Shinto shrine was built in
A Glimpse of History Koch’s honor. During Kitasato’s later years, he played an important role
Endospores of Clostridium tetani, the bacterium that causes tetanus (lockjaw), in establishing laws regulating health practices in Japan. When he died at
are found in soil and dust—virtually everywhere. This disease used to be com- age 75, a shrine in his honor was erected next to Koch’s.
mon before its cause and pathogenesis were understood, and it often ended in

M
an agonizingly painful death. Dr. Shibasaburo Kitasato (1856–1931), working ost people occasionally suffer wounds that cause breaks
on tetanus in Robert Koch’s laboratory in Germany, was the first to discover in the skin or mucous membranes. Microorganisms orig-
that C. tetani is an obligate anaerobe. This critical information helped him inating from the environment or the object causing the
develop a method to grow the bacterium in pure culture, an essential step wound almost always contaminate these injuries. Infection may
toward characterizing a pathogen and learning how it causes disease. result, depending on several factors, including (1) virulence of the
Kitasato showed that laboratory animals injected with C. tetani microbes; (2) number of microbial cells in the wound; (3) status of
developed tetanus. He was puzzled, however, by a surprising find-
the host’s immune system; and (4) the type of wound, especially
ing: Although the animals died of generalized disease, there were no
whether the tissues are crushed or contain foreign matter. Wounds
C. tetani cells anywhere other than the injection site. By doing experi-
ments in which he injected the tails of mice and then removed the inocu- that contain foreign materials such as dirt, leaves, bits of rubber,
lated tissue at hourly intervals, he showed that the animals developed or cloth usually become infected and do not heal until this matter
tetanus only if the bacteria remained in the animals for more than an is removed. Such wounds often provide places for microorgan-
hour. He also showed that the organisms stayed at the site of inoculation; isms to multiply out of the reach of phagocytes and other immune
at no time were they found in the rest of the body. Kitasato reasoned that defenses. In some cases, foreign materials create surfaces for
something other than bacterial invasion was causing the disease. biofilm development. They may also reduce available O2, thereby
While Kitasato was working with tetanus, another scientist, Emil inhibiting phagocytic function and allowing the growth of anaero-
von Behring, was busy investigating how Corynebacterium diphtheriae bic pathogens. Clean wounds often heal without treatment despite
caused the disease diphtheria. Together, Kitasato and von Behring showed microbial colonization, but sometimes even a minor wound can
that toxins produced by the bacteria caused both diseases. The concept that
result in a severe, or possibly fatal, infection.
a bacterial toxin could cause disease was an extremely important advance
in the understanding and control of infectious diseases. MicroByte
Kitasato published his studies in 1890 and 2 years later returned to Costs for treating postoperative wound infections are almost
Japan. The Japanese government did not support basic research at that $1.5 billion per year in the United States.
time, so Kitasato established his own institute for infectious diseases

548
 Part IV Infectious Diseases 549

23.1 ■ Anatomy, Physiology, ■ Gunshot wounds: Caused by bullets or other projectiles.

and Ecology ■ Burns: Caused by heat (thermal burns), electricity, chemi-
cals, radiation, or friction.
Learning Outcomes Wounds expose tissue components normally protected by skin or
1. Name three tissue components exposed by wounds to which mucous membranes, providing surfaces to which pathogens can
pathogens specifically attach. attach and then colonize. These tissue components include collagen,
2. Describe the beneficial and harmful aspects of abscess formation. fibronectin, fibrinogen, and fibrin. Collagen is a fibrous material—
the main supportive protein of skin, tendons, scars, and other body
Wounds vary in their characteristics, severity, and associated structures. Fibronectin is a fibrous glycoprotein that occurs both as
risks. The general categories of wounds include the following: a circulating form and as a component of tissue, where it binds cells
and other tissue substances together. Fibrinogen is a blood protein;
■ Incisions: Produced by a knife or other sharp object. when a wound occurs, this protein is converted to fibrin, which
■ Punctures: Result from penetration by a small sharp object, forms clots in the damaged vessels. The clots stop the flow of blood
such as a needle or a nail. as the first step in the wound repair process.
■ Lacerations: Occur when the tissue is torn. Wound healing begins with the outgrowth of connective
tissue cells (fibroblasts), and capillaries from the surfaces of
■ Contusions: Produced by a blow that crushes tissue. the wound, producing a red, translucent fibrous material called
■ Abrasions: Occur when the epidermis is scraped off. granulation tissue. In clean wounds, granulation tissue fills the
space created by the wound. This
Wound Blood clot tissue shrinks and is converted to
collagen, a component of scar tissue
Epidermis that is eventually covered by skin
or mucous membrane (figure 23.1).

Macrophages
Dermis
Fibroblast
Neutrophils

Neutrophils

1 Cut blood vessels bleed into the wound. 2 Blood clot forms in wound, and
Fibrin forms clots in severed capillaries. phagocytes destroy microbes.

Blood clot Scab

Macrophages
Regenerated
epithelium
Granulation (epidermis)
tissue
Regrowth of Scar tissue
blood vessel

Fibroblast Fibroblast

FIGURE 23.1 The Process

of Wound Repair
3 Wound fills with granulation tissue 4 Fibroblasts secrete collagen, forming scar tissue.
and blood vessels regrow. Collagen contracts and epithelium regenerates. ? What is the function of
granulation tissue?
550 Chapter 23 Wound Infections

Wound Abscesses often ineffective against microorganisms in the abscess because

the microbes stop multiplying, and most antimicrobial medica-
An abscess (figure  23.2) is a localized collection of pus sur-
tions work against actively dividing cells. Abscesses usually must
rounded by inflamed body tissue. The pus—a thick yellowish
therefore burst to a body surface or be drained surgically to be
fluid—is composed of living and dead leukocytes, tissue debris,
cured. antimicrobial medications, p. 457
and proteins. Abscesses form as a result of the body’s immune
defenses and usually indicate an infection. Although an abscess
helps localize the infection and prevents its spread, it also indi- Anaerobic Wounds
cates a potentially serious situation. If some cells of the pathogen An important feature of many wounds is that they are relatively
escape the abscess, they can enter the blood or lymph, leading to anaerobic, which allows the growth of obligate anaerobes such
infection in other parts of the body. as Clostridium tetani. Wounds that are likely to be anaerobic
The very nature of abscesses makes them difficult to treat. are those that have extensive tissue damage, are contaminated
They have no blood vessels, because the developing pus pocket with dirt or are small in diameter but deep, such as punctures.
destroys or pushes them aside, and adjacent blood vessels are Punctures may have foreign material and microorganisms forced
often blocked by clots. This lack of blood circulation makes it dif- deep into the tissues. Anaerobic conditions are also often created
ficult for antimicrobial drugs to reach the infected site. Even if the when different microbial species grow in a wound (polymicrobial
drugs do enter the site, the chemical nature of pus interferes with infections), because facultative anaerobes use up the available O2,
the action of some antibiotics. In addition, antimicrobial drugs are converting it to water as they respire.

FIGURE 23.2 Abscess Microorganisms

Formation
? What is the composition Epidermis
of pus?

Dermis

Capillary
Neutrophils

1 Microorganisms enter the tissue from 2 Blood vessels dilate, and leukocytes migrate
a wound or from the bloodstream. to the area of the developing infection.

Pus

Blood clots

Blood clots

3 Pus forms and an abscess develops; 4 Buildup of pressure causes the abscess to expand in
clotting occurs in adjacent blood vessels. the direction of least resistance; if it reaches a body
surface, it may rupture and discharge its contents.
 Part IV Infectious Diseases 551

MicroAssessment 23.1
Wounds can be classified as incisions, punctures, lacerations,
contusions, abrasions, gunshots, or burns. Wounds expose tissue
components to which pathogens can attach. Healing involves the
outgrowth of fibroblasts and capillaries from the sides of the wound
to produce granulation tissue that fills the wound. Abscess formation
provides a way of isolating infections and preventing their spread.
Anaerobic conditions in wounds are created by the presence of dead
tissue and foreign material.
1. Name and describe two substances in wounds to which
pathogens attach.
2. Give two reasons why an abscess might not respond to antibiotic
treatment.
3. Why is it important that the granulation tissue shrinks after
it is formed? +

23.2 ■ Common Bacterial

Infections of Wounds
Learning Outcomes
3. Give distinctive characteristics of three common wound infections
caused by bacteria that grow aerobically.
FIGURE 23.3 Surgical Wound Infection Due to
4. Discuss the significance of fibronectin binding by S. epidermidis. Staphylococcus aureus
? Why do infected surgical wounds sometimes split open?
If a wound becomes infected, several serious consequences are
possible. These include (1) delayed healing, (2) formation of
abscesses, and (3) spread of the bacteria or their toxins to other
areas of the body. Infected surgical wounds often split open as
swelling causes the stitches to pull through tissues weakened
by the infection. The infection can spread to devices such as an
artificial hip or knee, which may then have to be removed before Signs and Symptoms
the infection can be eliminated. Table 23.1 summarizes the char- Staphylococcus species are pyogenic, meaning that they cause the
acteristics of the leading causes of wound infections. production of pus (pyo means “pus” and genic means “generat-
ing”). Staph infections are usually characterized by an inflamma-
tory reaction, with swelling, redness, and pain. Fever occurs if the
Staphylococcal Wound Infections infected area is large or if the infection has spread to the blood or
Staphylococcus species, common inhabitants of the nostrils and lymph.
skin, are the leading causes of wound infections (figure 23.3). Of Toxic shock syndrome can occur if the wound is infected
the 30 or more recognized species, only two cause most human with a toxin-producing staphylococcal strain. Signs and symptoms
wound infections—Staphylococcus aureus and Staphylococcus of this include high fever, muscle aches, and a life-threatening
epidermidis. The more important of these two, S. aureus, was drop in blood pressure and shock. Sometimes the infected person
covered extensively in chapter 22. the genus Staphylococcus, p. 273 will also have a rash and diarrhea. staphylococcal toxic shock
bacterial skin diseases, p. 523 syndrome, p. 619

TABLE 23.1 Leading Causes of Wound Infections

Causative Organism Characteristics Consequences

Staphylococcus aureus Gram-positive cocci in Delayed healing; abscess formation; extension into tissues, artificial devices,
clusters, coagulase-positive or bloodstream; some strains can cause toxic shock syndrome
Streptococcus pyogenes Gram-positive cocci in Same as above, except some strains can cause “flesh-eating” necrotizing
chains; Lancefield group A fasciitis
Pseudomonas aeruginosa Gram-negative rod, green Delayed healing; abscess formation; extension into tissues, artificial devices,
pigment or bloodstream; septic shock
552 Chapter 23 Wound Infections

Causative Agents abscesses in the heart, bones, or other tissues. Some S. aureus
Staphylococcus aureus and S.  epidermidis are Gram-positive strains produce superantigens that can enter the circulation.
cocci that grow in clusters (figure 23.4). These facultative anaer- These exotoxins react with helper T cells, activating them and
obes are quite hardy, which is not surprising because they have causing them to release large amounts of cytokines that lead to
evolved to thrive on skin, which is dry and salty. They survive toxic shock. superantigens, p. 393 cytokines, p. 341

well in the environment and are easily transferred from person to

Staphylococcus epidermidis S.  epidermidis is not particularly
person. facultative anaerobe, p. 90
virulent and cannot invade healthy tissues. However, the bacterium
The coagulase test is used to distinguish S. aureus from other
often causes minor abscesses around the stitches used in surgery.
staphylococci. S. aureus is often referred to as “coag-positive
It also adheres to and then colonizes medical devices, including
staph” because it makes coagulase, whereas the other staphy-
indwelling catheters and artificial joints. It can do this because it
lococcal species are collectively referred to as “coag-negative
binds to fibronectin, the blood protein that quickly coats surgical
staph.” Of the coag-negative staphylococci, S. epidermidis is the
implants in the body. The bacterial cells may then produce a slime
most common cause of healthcare-associated infections, includ-
layer or glycocalyx, a critical step in biofilm formation.
ing those of surgical wounds. However, S. aureus causes serious
Biofilms are a serious problem for several reasons. Diffusion
wound infections much more commonly than S. epidermidis.
of antibacterial medications into them is slow and inefficient. Even
Healthcare-associated infections, p. 449 coagulase, p. 526
if the medication does enter the biofilm, it may be ineffective
because bacteria within the biofilm are often metabolically inac-
Pathogenesis
tive. In addition, bacteria in biofilms can come loose, and are then
Staphylococcus aureus S. aureus produces multiple virulence carried by the bloodstream to the heart and other tissues. In people
factors that act together in the disease process (see table 22.3). with a compromised immune system, such as those with cancer,
These virulence factors are covered extensively in chapter 22 (skin AIDS, or diabetes mellitus, this can result in subacute bacterial
infections), but it is important to recognize that those same fac- endocarditis or multiple tissue abscesses, which require surgical
tors play a critical role in wound infections as well. For example, treatment. In healthy people, wound infections by S. epidermidis
clumping factor and other proteins allow the cells to attach to clots are usually cleared by host defenses without additional treatment.
and tissue components, an initial step in colonization. Lipases, glycocalyx, p. 62 biofilm, p. 84 bacterial endocarditis, p. 672
proteases, and hyaluronidases together cause tissue damage.
Capsules, coagulase, and protein A protect the cells from attack by MicroByte
the complement system, phagocytes, and antibodies. S.  aureus It takes more than 100,000 S. aureus cells injected into skin to cause
pathogenesis, p. 524 an abscess, whereas only 100 injected into a suture site to do the same.
Immunity to staphylococcal infection is generally weak or
non-existent, probably because the organism so effectively evades
the immune defenses. However, some protein A is released from
Epidemiology
the bacterial surface, and it reacts with circulating antibodies. The S. aureus carriers are at an increased risk for surgical wound infec-
resulting immune complexes can activate the complement sys- tions caused by this species. Other factors that increase a person’s
tem, probably contributing to the intense inflammatory response risk include advanced age, poor general health, immunosuppres-
and accumulation of pus that characterize S. aureus infections. sion, prolonged preoperative hospital stay, and an infection at
complement system, p. 344 another site. Additional information about the epidemiology of
Staphylococcus aureus infections may cause systemic com- S. aureus is discussed in chapter 22. S. aureus epidemiology, p. 526

plications. Bacteria growing in a wound can spread, leading to S. epidermidis is found on the skin and mucous membranes
of most people, residing as part of their normal microbiota. It is an
opportunist that can cause disease in individuals with a compro-
mised immune system. opportunist, p. 382

Treatment and Prevention

Leukocytes
Treating staphylococcal infections can often be difficult because of
widespread antibiotic resistance. Methicillin-resistant S. aureus
(MRSA) is a serious problem in wound infections. This organism
Bacteria is extremely difficult to treat, being resistant to multiple β-lactam
antibiotics, including methicillin, penicillin, nafcillin, and oxacil-
lin. MRSA infections may be subcategorized into healthcare-
associated (HA-MRSA) infections and community-acquired
(CA-MRSA) infections. Both HA-MRSA and CA-MRSA are
resistant to multiple antibiotics, although CA-MRSA is more
FIGURE 23.4 Staphylococcus aureus in Pus susceptible than HA-MRSA and can be treated successfully with
? What does the name Staphylococcus indicate about the typical sulfa drugs, tetracyclines, and clindamycin in about 75% of cases.
growth arrangement of the bacterial cells? HA-MRSA is resistant even to these and is often susceptible only
 Part IV Infectious Diseases 553

to vancomycin. Vancomycin-intermediate S. aureus (VISA) and person develops fever and confusion. The overlying skin becomes
vancomycin-resistant S. aureus (VRSA) have emerged, making stretched and discolored because of the swelling. Unless treatment
treatment of these infections extremely difficult. New medications is started quickly, shock and death usually follow in a short time.
belonging to the oxazolidinones class may be effective in treating
them. antibacterial resistance, p. 471 MRSAs, p. 473 VISA and VRSA, Causative Agent
p. 473 Streptococcus pyogenes is a β-hemolytic, Gram-positive, chain-
To reduce the chance of infection, wounds should be thor- forming, aerotolerant organism, with Lancefield group A cell
oughly cleaned, removing any dirt or dead tissue. Clean, deep wall polysaccharide and a nonantigenic capsule of hyaluronic
wounds and surgical wounds should be quickly closed by sutures acid. Strains of S. pyogenes that cause invasive disease are more
to help avoid infection. Surgical wound infections can be reduced virulent because they produce various enzymes and toxins that
by half if the patient is given an effective anti-staphylococcal cause severe tissue damage (covered next). aerotolerance, p.  90
medication immediately before surgery. For unknown reasons, β-hemolysis, p. 96 pyrogens, p. 351
the infection rate is actually increased if the medication is given
more than 3 hours before or 2 hours after the surgical incision. Pathogenesis
The pathogenesis of Streptococcus pyogenes has been covered in
Group A Streptococcal detail in chapter 21. Like Staphylococcus aureus, S. pyogenes has
a fibronectin-binding protein (F protein) that helps colonization
“Flesh-Eating Disease”
of wounds (see table 22.5). In necrotizing fasciitis, S. pyogenes
Streptococcus pyogenes is another common cause of wound destroys the subcutaneous fatty tissue and fascia, the bands of
infections. S. pyogenes infections have generally been easy to fibrous tissue that underlie the skin and surround muscle and
treat because all known strains of the organism are still suscep- body organs. In some cases, the organism destroys the muscle
tible to penicillin. Occasionally, however, the infections can tissue itself. It does this damage by producing a variety of destruc-
progress rapidly, even leading to death despite antimicrobial tive enzymes such as streptokinases (break down blood clots),
treatment. These more severe infections are called “invasive” hyaluronidases (break down connections between cells), deoxyri-
because they spread into tissues and organs, causing pneumo- bonucleases (break down DNA), and streptolysins (break down
nia, meningitis, puerperal fever, fasciitis—inflammation of the red blood cells and neutrophils). Strains that cause necrotizing
fascia that surround muscles and body organs—and streptococ- fasciitis also produce exotoxin A, a superantigen that causes toxic
cal toxic shock. This section will focus on necrotizing fasciitis shock, and exotoxin B, a protease that destroys tissue. As the tis-
(“flesh-eating disease”), a rare but serious complication of sues break down, osmotic pressure increases, and fluid moves into
S. pyogenes infection (figure 23.5). Streptococcus pyogenes, p. 487 the area, causing intense swelling. The organisms multiply in the
streptococcal toxic shock, p. 553 dead tissue, using the breakdown products as nutrients.
As the bacteria grow, they shed M protein, a surface protein
Signs and Symptoms which binds to fibrinogen. The M protein–fibrinogen complexes
Signs and symptoms of necrotizing fasciitis appear suddenly and bind to neutrophils, causing them to release strong inflammatory
are very serious. Severe pain develops at the site of the wound, molecules called heparin-binding proteins that increase the vascu-
which sometimes can be so minor that no break in the skin is lar permeability in the host. The blood vessels leak fluid, resulting
even seen. Within a short time, swelling occurs, and the injured in a massive, life-threatening drop in blood pressure and shock.
M protein also helps the organism avoid phagocytosis by breaking
down complement component C3b.
In many cases, S. pyogenes also releases a variety of
streptococcal pyrogenic exotoxins (SPEs), which are superan-
tigens. These enter the bloodstream, activating T cells and
causing them to release large amounts of cytokines. The high
levels of cytokines causes symptoms associated with strepto-
coccal toxic shock syndrome (TSS) including fever, nausea,
vomiting, rash and sometimes shock and death. Table 22.5
compares and contrasts the pathogenesis of S. pyogenes and
S. aureus.

Epidemiology
Cases of flesh-eating disease in the United States are generally
sporadic. Of the deaths caused by invasive S. pyogenes infections
in the United States yearly, less than 2% are due to necrotizing fas-
FIGURE 23.5 Individual with Streptococcus pyogenes ciitis. Underlying conditions that increase the risk of necrotizing
“Flesh-Eating Disease” (Necrotizing Fasciitis) fasciitis and other invasive S. pyogenes infections include diabe-
? Why does necrotizing fasciitis require immediate surgery? tes, cancer, alcoholism, AIDS, recent surgery, abortion, childbirth,
554 Chapter 23 Wound Infections

chickenpox, and injected-drug abuse. Invasive infections seldom

occur in healthy individuals with minor injuries.

Treatment and Prevention

The toxins produced by S. pyogenes spread with such speed that
immediate surgery is often essential to reduce the pressure of the
swollen tissue and to remove dead tissue. Amputation is some-
times necessary to quickly remove the source of toxins. Penicillin
is effective for treating early infection, but it has little or no
effect on streptococci in necrotic tissue and no effect on toxins.
Therefore surgery remains the best treatment.
There are no proven preventive measures for S. pyogenes
fasciitis, although M protein vaccines are being tested.

(a)
Pseudomonas aeruginosa Infections
Pseudomonas aeruginosa, an opportunistic pathogen, is a major
cause of healthcare-associated infections. In hospitals, P. aeruginosa
is an important cause of lung infections and a common cause of wound
infections, especially of thermal burns. Burns have large exposed
areas of dead tissue that are not protected by normal body defenses
and are therefore ideal sites for infection by bacteria. Almost any
(b)
opportunistic pathogen can infect burns, but P. aeruginosa is among
the most common and most difficult to treat. Pseudomonas, p. 265
P. aeruginosa also occasionally causes community-acquired
infections. Such infections include skin rashes and external ear
canal infections from contaminated swimming pools and hot tubs,
and eye infections from contaminated contact lens solutions. Other
infections caused by this organism include those of foot bones
from stepping on sharp objects, heart valve infections in injected-
drug abusers, scarring infections from ear piercing, and biofilms
in the lungs of individuals with the inherited disease cystic fibrosis.

Signs and Symptoms

Signs and symptoms of Pseudomonas aeruginosa infection are
serious and include chills, fever, skin lesions, and shock, which
are caused by bloodstream invasion by this organism. A very
noticeable symptom of P. aeruginosa infection of burns and other
wounds is a characteristic green color, caused by water-soluble
10 μm
pigments produced by the organisms (figure 23.6a; see also figure (c)
11.12). These pigments include fluorescent yellow pyoverdin and FIGURE 23.6 Pseudomonas aeruginosa (a) Extensive burn
blue pyocyanin, which together produce the green color. infected with P. aeruginosa. (b) Culture. (c) P. aeruginosa has a single
polar flagellum.
Causative Agent ? What causes the green discoloration of the wound and the culture
Pseudomonas aeruginosa is a motile Gram-negative rod with a medium?
single polar flagellum (figure 23.6c). It is found in a wide variety
of environments such as soil and water, where it grows easily and production of two extracellular proteins—exotoxin A and exoen-
fast. The bacterium is classified as an aerobe. However, it also zyme S. Exotoxin A stops host cell protein synthesis. Exoenzyme
respires anaerobically in the absence of O2 if nitrate is present. The S is a phospholipase that acts synergistically with a protease called
ability of P. aeruginosa to respire anaerobically is critical in the lecithinase to hydrolyze lecithin, an important lipid component of
development of biofilms by this organism. anaerobic respiration, cell membranes. Destruction of lecithin causes membrane disrup-
p. 134 the genus Pseudomonas, p. 265 tion and cell death.
The pigment pyocyanin produced by P.  aeruginosa may
Pathogenesis further add to pathogenicity. A derivative of this pigment acts as
Pseudomonas  aeruginosa infection of burns and other wounds a siderophore, helping the pathogen acquire iron from the envi-
causes additional tissue damage, delays healing, and increases the ronment, thereby inhibiting competing bacteria by reducing their
risk of septic shock. Virulence of the organism depends mainly on available iron. This pigment also impairs the function of human
 Part IV Infectious Diseases 555

nasal cilia and disrupts respiratory epithelium. No role in virulence Wounds are often anaerobic and may be colonized by certain
is known for the pigment pyoverdin. siderophores, p. 386 strictly anaerobic species of bacteria. This section describes three
distinctive diseases that result from anaerobic bacterial wound
Epidemiology infections: lockjaw, gas gangrene, and lumpy jaw.
Pseudomonas aeruginosa is widespread in nature. The organ-
ism can grow in most places where there is moisture, including
soaps, ointments, eyedrops, contact lens solutions, cosmetics, Tetanus (“Lockjaw”)
disinfectants, swimming pools, hot tubs, and even distilled water. Tetanus is often fatal. Fortunately, it is uncommon in economi-
P. aeruginosa is introduced into hospitals on ornamental plants, cally advanced countries. Exposure to the causative organism
flowers, and produce. For this reason, visitors are not allowed to cannot be avoided because it produces endospores that are wide-
take flowers or fruit into hospital burn wards or intensive care spread in dust and dirt, frequently contaminating clothing, skin,
units. It can also be found on many kinds of hospital equipment, and wounds. Even a minor wound in a non-immunized person
the inner soles of shoes, and in illegal injectable drugs, all of can result in tetanus if the wound provides conditions that allow
which have been sources of serious infections. germination of the spores. endospores, p. 67

Treatment and Prevention

Signs and Symptoms
Pseudomonas aeruginosa infections are treated with antimicrobial
Tetanus is characterized by continuous, painful, and uncontrol-
medications. Established infections, however, are very difficult to
lable cramplike muscle spasms that are usually generalized
treat because P. aeruginosa is resistant to a wide range of antibi-
but may be limited to one area of the body (figure 23.7). The
otics. Only a few antibiotics are effective against this pathogen,
spasms often begin with the jaw muscles, giving the disease the
including fluoroquinolones, gentamicin, and imipenem—and even
common name “lockjaw.” The early symptoms include restless-
these drugs may not be effective against all strains. Antibiotic
ness, irritability, difficulty swallowing, contraction of the jaw
sensitivity tests are done to determine the most appropriate medi-
muscles, and sometimes seizures. As more muscles go into sus-
cation for treatment. For systemic infection, antibacterial medica-
tained contraction (called tetany), breathing becomes difficult,
tions must usually be given intravenously in high doses.
abnormal heart rhythms may occur, and in some cases bones can
P.  aeruginosa infections can be prevented by eliminating
fracture. After a period of almost unbearable pain, the infected
possible sources of the bacterium and prompt care of wounds.
person often dies of pneumonia or from lung damage caused
Removing dead tissue from burn wounds, followed by application
by regurgitation of stomach contents into the lung. In addition,
of an antibacterial cream, also helps prevention of infection by
because people being treated for tetanus remain in the hospital
this organism.
for long periods of time, healthcare-associated infections can
develop.
MicroAssessment 23.2
Staphylococcus aureus is the most important cause of wound
Causative Agent
infections because it is commonly carried by humans, it transfers
easily from one person to another, and it has multiple virulence Tetanus is caused by Clostridium tetani, an anaerobic, spore-
factors. Staphylococcus epidermidis forms biofilms on foreign forming, Gram-positive, rod-shaped bacterium (figure  23.8).
materials, protecting the bacteria from body defenses and antibacterial
medications. Flesh-eating strains of Streptococcus pyogenes are
uncommon but can cause life-threatening disease. Pseudomonas
aeruginosa, a pigment-producing organism, is widespread in the
environment and is a major cause of healthcare-associated infections.
4. Why are antibacterial medications not effective for treating
flesh-eating disease (necrotizing fasciitis)?
5. Why do wound infections caused by Pseudomonas aeruginosa
sometimes produce green pus?
6. Why is it not surprising that staphylococci are the most common
cause of wound infections? +

23.3 ■ Diseases Due to Anaerobic

Bacterial Wound Infections
Learning Outcomes
5. Describe the conditions that lead to the development of anaerobic
wound infections.
6. Discuss why it is difficult to treat wounds infected with toxin- FIGURE 23.7 Infant with Neonatal Tetanus
producing bacteria.
? How does a newborn infant contract tetanus?
556 Chapter 23 Wound Infections

from inhibitory neurons, blocking their action and causing the mus-
cles to contract without control (figure 23.9). A-B toxins, p. 392
The toxin generally spreads across the spinal cord to the side
opposite the wound and then downward, typically causing spastic
muscles to first appear on the side of the wound, then on the oppo-
site side, and then downward, depending on the amount of toxin.
Tetanospasmin released from the infected wound often enters the
bloodstream, which carries it to the central nervous system. In
these cases, inhibitory neurons of the brain are first affected, and
the muscles of the jaw are among the first to become spastic.

Epidemiology
Clostridium tetani occurs not only in dirt and dust, but also in the
gastrointestinal tract of humans and other animals that have eaten
foods contaminated with its spores. Therefore, fecal contamina-
Endospores 5 μm tion is a potential source of infection. Many cases of tetanus result
FIGURE 23.8 Clostridium tetani Terminal endospores are from puncture wounds, which occur in stepping on a nail, body
characteristic of this species. piercing, tattooing, animal bites, splinters, injected-drug abuse,
and insect stings. Cases can also occur following surgery. Surface
? How are the endospores shown here different from endospores
of other clostridial species? abrasions and burns can result in tetanus if the wound is anaerobic
because of dirt or dead tissue.
In economically developed countries, people who get tetanus
It has two characteristic features: (1) a spherical endospore have either never been immunized or are no longer immune because
that forms at the end of the cell, and (2) swarming growth that they received their last vaccine booster shot more than ten years
quickly spreads over the surface of solid media. Identification of ago. In less economically advanced countries, tetanus occurs more
C. tetani depends on characterizing its plasmid-encoded toxin, frequently because vaccination is not available, and because of
although tetanus is typically diagnosed by signs and symptoms. improper wound care. In some parts of the world, babies commonly
exotoxins, p. 391 Clostridium, p. 258 plasmids, p. 208 die of neonatal tetanus as a result of their umbilical cord being cut
with instruments contaminated with C. tetani (see figure 23.7).
Pathogenesis
Clostridium tetani is not invasive, and colonization is gener- Treatment and Prevention
ally localized to a wound. Its pathologic effects are caused by Tetanus is treated by injecting the affected person with human
tetanospasmin, an exotoxin released from multiplying vegetative tetanus immune globulin (TIG), a preparation of antibodies to
cells. Tetanospasmin is an A-B toxin. The B portion attaches to the tetanus toxin. The antibodies bind to toxin molecules not
receptors on motor neurons, which then take up the A portion by yet attached to nerve cells, thereby neutralizing their effects and
endocytosis. The toxin is carried to the neuron cell body in the providing passive immunity. TIG, however, cannot neutralize
spinal cord. There, the motor neuron contacts other neurons that tetanospasmin that is already attached to nerve tissue and it does
normally control its action by means of chemicals called neu- not repair nerve damage that has already occurred. In this case,
rotransmitters. Some of these other neurons stimulate the motor the affected person is given muscle relaxants and supportive care,
nerve cell, causing
muscle contraction, Normal (Flexion) Normal (Extension) Tetanus
while others make
the motor neuron
resistant to stimu-
lation, inhibiting
muscle contraction. Motor Motor
neuron neuron
Tetanospasmin pre- stimulated; inhibited;
vents the release of this muscle this muscle
the neurotransmitter contracts. relaxes.

FIGURE 23.9 Motor Motor

Tetanus and neuron neuron
Inhibitory Neuron Both
inhibited; stimulated; muscles
Function this muscle this muscle contract and
relaxes. contracts.
? How does arm cannot
tetanospasmin cause move.
muscle contraction?
 Part IV Infectious Diseases 557

Tetanus can easily be avoided by vaccination

TABLE 23.2 Tetanus Prevention in the Management of Wounds
with tetanus toxoid, which is inactivated tetano-
CLEAN MINOR WOUNDS ALL OTHER WOUNDS spasmin. Immunization with tetanus toxoid in
Immunization combination with diphtheria toxoid and acellular
History Toxoid TIG Toxoid TIG
pertussis vaccine, called DTaP, is recommended in
Unknown, or fewer than Yes No Yes Yes children under the age of 7. DTaP is given at 2, 4,
three injections 6, and 18 months of age, with a booster dose when
Fully vaccinated (three or children enter school. Additional booster doses of
more injections of toxoid) tetanus toxoid are given to people at 10-year inter-
■ 5 years or less since No No No No vals to maintain an adequate level of immunity.
last dose Currently DTaP is recommended for booster doses
■ 5 to 10 years since last No No Yes No in people ages 10 to 64 years. Updating of booster
dose doses of toxoid more frequently than every 10
■ More than 10 years since Yes No Yes No years can be dangerous because there is risk of an
last dose allergic reaction in individuals who already have
large amounts of antibodies against it. Individuals
who have recovered from tetanus are not immune
including being placed on a ventilator if needed. Over time, the to the disease and must be immunized. People who have been injured
affected nerves repair themselves. TIG, p. 420 or burned or are planning surgery should receive a booster shot of
In addition to TIG treatment, the wound is thoroughly cleaned of the tetanus vaccine. Table 23.2 summarizes prevention of tetanus by
all dead tissue and foreign material that could cause anaerobic condi- wound management. toxoid, p. 423
tions. An antibacterial medication such as metronidazole is given to Neonatal tetanus can be prevented by vaccinating pregnant
kill any actively multiplying clostridia, preventing the production of women. Their infants are then protected by maternal antibodies
more tetanospasmin. The person is also given tetanus vaccine, at a crossing the placenta. Table 23.3 describes the main features of
different site from the TIG injection. tetanus.

TABLE 23.3 Tetanus (“Lockjaw”)

1 Clostridium tetani spores from Symptoms Restlessness, irritability, difficulty

dust or dirt enter a wound. 3 swallowing; muscle pain and spasm in
2 jaw, abdomen, back, or entire body
In anaerobic wounds, the
spores germinate, and Incubation period 3 days to 3 weeks; average 8 days
vegetative bacteria release an 4 Causative agent Clostridium tetani, an anaerobic, spore-
exotoxin called tetanospasmin. forming, Gram-positive rod
3 5
Tetanospasmin is carried to
Pathogenesis Tetanus results from tetanospasmin, an
the central nervous system by exotoxin produced by the bacterium. The
motor nerve axons or by the 3
toxin is carried to the brain and spinal
bloodstream. 6 cord by motor nerve axons or circulating
4 The toxin prevents any blood; toxin acts against nerve cells that
inhibitory neurons it reaches normally inhibit muscle contraction.
from functioning. 3 Other nerves that normally cause muscle
contraction then act unopposed, causing
5 The corresponding neurons,
muscle spasms.
which cause muscles to
contract, act unopposed by Epidemiology Organisms common in soil; spores
inhibitory neurons. contaminate wounds, germinate in those
having anaerobic conditions, particularly
6 The result is a sustained,
dirty or puncture wounds.
painful cramplike muscle
spasm. Treatment and Treated with metronidazole, tetanus
prevention antitoxin. Immunization of children at
3 ages 2 months, 4 months, 6 months,
18 months; booster dose at time of
entering school and at 10-year intervals
after that; tetanus immune globulin
(TIG), cleaning wound.

1
558 Chapter 23 Wound Infections

MicroByte Although C. perfringens is a spore-forming organism, it usually

World Health Organization education initiatives in developing does not produce spores in wounds or cultures.
countries have reduced the cases of neonatal tetanus by almost 75%
in the last 20 years Pathogenesis
Two main factors lead to the development of gas gangrene: (1) the
Clostridial Myonecrosis presence of dirt and dead tissue in the wound, and (2) long delays
before the wound is treated. Clostridium perfringens is unable to
(“Gas Gangrene”) infect healthy tissue but grows easily in poorly oxygenated and
Before antibiotics were discovered, clostridial myonecrosis (com- dead tissues that provide it with a source of growth factors and
monly called “gas gangrene”) killed many soldiers wounded in amino acids. It releases α-toxin, an enzyme that destroys lecithin
wars. Endospores of Clostridium perfringens, the bacterium that in host cell membranes, leading to cell lysis. The toxin diffuses
can cause this disease, are found in soil and dust everywhere, from the area of infection, killing leukocytes and tissue cells.
and frequently can be isolated from wounds. However, infection Several enzymes produced by the pathogen, including collagenase
with C. perfringens rarely leads to gas gangrene, because like and hyaluronidase, break down the host tissues. The organisms
C. tetani, the organism grows only in anaerobic conditions. Gas multiply, using the tissue breakdown products, and release hydro-
gangrene occurs mainly in neglected wounds with fragments of gen and carbon dioxide. These gases accumulate in the tissue
bone, foreign material, and serious tissue damage. It also occurs and cause a rise in pressure, which leads to further spread of the
occasionally in surgical sites, especially in people with underlying infection. Without quick surgical treatment, massive amounts
diseases. of α-toxin diffuse into the bloodstream and destroy red blood
cells, tissue capillaries, and other structures throughout the body.
Signs and Symptoms C. perfringens infections are generally not dangerous until they
Gas gangrene signs and symptoms appear suddenly and dra- invade muscle, but if they do, severe toxicity occurs quickly for
matically. They include severe pain that quickly increases in the unknown reasons. It is not reversed by treatment with antibody to
infected wound, followed by swelling in the area, and a thin, α-toxin. growth factors, p. 93
bloody or brownish fluid that leaks from the wound. This fluid
may look frothy because of gas bubbles released by the organism. Epidemiology
The overlying skin becomes stretched tight and mottled with black Clostridium perfringens is widespread in soil and is common in
(figure  23.10). Although seriously ill, the victim remains quite the feces of many animals and humans. It is sometimes found in
alert until late in the illness when, near death, he or she becomes the vagina of healthy women. Besides neglected trauma wounds
delirious and goes into a coma. and occasional surgical wounds, gas gangrene of the uterus is
fairly common after self-induced abortions, and occasionally it
Causative Agent can occur after miscarriages and childbirth. In other cases, dis-
Several species of Clostridium can cause gas gangrene when they eases such as arteriosclerosis or diabetes—which lead to poor
invade injured muscle, but by far the most common is C. perfringens, oxygenation of tissue from restricted blood flow—are predispos-
an encapsulated Gram-positive anaerobic toxin-producing rod. ing factors. Cancer patients also have increased susceptibility to
gas gangrene.

Treatment and Prevention

Treatment of gas gangrene includes prompt surgical removal
of all dead and infected tissues, and may require amputations.
Antibiotics such as penicillin are given to help stop bacterial
growth and toxin production. However, because the drugs do not
diffuse well into large areas of dead tissue and do not inactivate
toxin, they are of little use in treating the disease. Hyperbaric oxy-
gen treatment is sometimes used, in which the patient is placed
in a special chamber and breathes pure O2 under increased pres-
sure. The pressure treatment inhibits growth of the clostridia, and
so stops the release of toxin. The high levels of O2 also improve
oxygenation of injured tissues, lessening anaerobic conditions.
Hyperbaric treatment is not always successful and may lead to
FIGURE 23.10 Individual with Gas Gangrene (Clostridial other complications.
Myonecrosis) Fluid leaking from the involved area typically shows There is no available vaccine for gas gangrene. The disease
bits of muscle digested by Clostridium perfringens. Leukocytes are can be prevented by prompt cleaning and removal of dead tissue
absent. from wounds (debridement). Table 23.4 describes the main fea-
? How is gas gangrene treated? tures of this disease.
 Part IV Infectious Diseases 559

TABLE 23.4 Clostridial Myonecrosis (“Gas Gangrene”)

1 Clostridium perfringens Signs and Severe pain; gas and fluid seep from
spores enter a wound with Symptoms wound, blackening of overlying skin;
two essential characteristics: shock and death commonly follow
dead tissue and anaerobic Incubation Usually 1 to 5 days
conditions. period
2 The spores germinate, and the
Causative Usually Clostridium perfringens; other
vegetative bacteria multiply in
agent clostridia less frequently
dead tissue, producing α-toxin.
Pathogenesis Organism grows in dead and poorly
3 α-toxin diffuses into normal
5 oxygenated tissue and releases α-toxin;
tissue and kills it. The infection
toxin kills leukocytes and normal tissue
spreads into the dead tissue,
cells by degrading lecithin in their cell
the bacteria using amino acids
membranes; involvement of muscle
and growth factors released
causes shock by unknown mechanism.
from the tissue by bacterial
enzymes. Epidemiology Wounds of war; dirt contamination
of wounds, tissue death, impaired
4 Swelling and gas produced by
circulation to tissue as in people with
fermenting amino acids and
diabetes or arteriosclerosis; self-induced
muscle glycogen aid rapid
abortions.
progress of the infection. 4
1 Treatment Treated by surgical removal of dirt and
5 Massive amounts of α-toxin are
and prevention dead tissues of primary importance;
produced and diffuse into the 2 hyperbaric oxygen of possible value;
bloodstream, destroying blood
antibiotics to kill vegetative C. perfringens
cells and other cells throughout
of marginal value. Prevented by prompt
the body.
cleaning and debridement of wounds; no
vaccine available.
3

Actinomycosis (“Lumpy Jaw”)

Many wound infections are caused by anaerobes other than
clostridia, frequently by members of the normal microbiota of the
mouth and intestine. The disease commonly called “lumpy jaw”
was originally thought to be a fungal disease, and was therefore
given the name actinomycosis. We now know that lumpy jaw is
not a mycosis, but its original name is still used.

Signs and Symptoms

Actinomycosis is characterized by slowly progressing, sometimes
painful, swellings under the skin that eventually burst and drain
pus. The openings usually heal, only to reappear at the same or
nearby areas days or weeks later. Most cases involve the area of
the jaw and neck and cause swellings and scars, leading to the
popular name lumpy jaw (figure 23.11). In some cases, the swell-
ings and drainage develop on the chest or abdominal wall, or in FIGURE 23.11 Actinomycosis (”Lumpy Jaw”)
the genital tract of women. ? Is actinomycosis caused by a fungus?
560 Chapter 23 Wound Infections

CASE PRESENTATION
A 63-year-old woman, healthy except for 1. Cultures of surfaces in the operating room susceptibility to infection—cancer in
mild diabetes, had her gallbladder surgi- grew large numbers of C. perfringens. one, and diabetes in the other. Moreover,
cally removed. The surgery went well, but 2. Unknown to the medical staff, a workman both had a recognized source for the
within 72 hours the surgical incision became had recently serviced a fan in the organism. Cultures of as many as 20%
swollen and pale. Within hours, the swollen ventilation system of the operating room, of diseased gallbladders are positive for
area widened and developed a bluish dis- and for a time air was allowed to flow into Clostridium perfringens, and the organism
coloration. The woman’s surgeon suspected the operating room, rather than out of it. is commonly found in large numbers in
gangrene. Antibiotic therapy was started, and 3. Heavy machinery was doing road repair the human intestine—a potential source in
she was rushed back to the operating room outside the hospital, creating clouds of the case involving removal of the bowel
where the entire swollen area, including the dust. malignancy.
repaired operative incision, was surgically 3. The surgeon thought that the infecting
removed. After that, the wound healed nor- As a result of these findings, the operating
organism might have come from the
mally, although she required a skin graft to room and its ventilating system were cleaned
patients themselves because such strains
close the large wound. Large numbers of and upgraded. No further cases of surgical
are often much more virulent than strains
Clostridium perfringens grew from the wound wound gangrene developed.
that live and sporulate in the soil. On the
culture. 1. Was the surgeon’s diagnosis correct? other hand, the gross contamination of the
Six days later, a 58-year-old woman had 2. Many other patients had surgery in the operating room, as revealed by the cultures
surgery in the same operating room for a same operating room. Why did only these of its surfaces, could indicate a very large
malignant tumor of the colon. The surgery two patients develop wound gangrene? infecting dose at the operative site, pos-
was performed without difficulty, but 48 hours sibly compensating for lesser virulence.
3. What could be done to help identify the
later she developed rapidly advancing swelling In addition, no further cases occurred after
source of the patients’ infections?
and bluish discoloration of her surgical wound. cleaning the operating room and fixing
As with the first case, gangrene was suspected the ventilation system. Unfortunately, in
and she was treated with antibiotics and surgi- Discussion
this case, no cultures of the removed
cal removal of the affected tissue. She also 1. Clostridium perfringens is commonly cul- gallbladder or bowel tumor were done,
needed a skin graft. Her wound culture also tivated from wounds without any evidence nor were the strains isolated from the
showed C. perfringens. of infection. However, in these cases, wounds and the environment compared.
Because the surgery department had never there was not only a heavy growth of the Comparing the antibiotic susceptibility,
had any of its patients develop surgical wound organism but also a clinical picture that toxin production, and other characteristics
infections with C. perfringens before this, suggested gangrene. The surgeon’s diag- of the different isolates could have helped
the hospital epidemiologist was asked to do nosis was correct. identify the source of the infections.
an investigation. Among the findings of the 2. In both cases, there was an underlying
investigation: condition that increased the two patients’

Causative Agent
Neutrophils
Most cases of actinomycosis are caused by Actinomyces israelii,
a Gram-positive, filamentous, branching, slow-growing anaerobic
bacterium. A number of similar species can cause the disease in ani- Radiating
mals and humans. Despite its name, this organism is not a fungus. filaments

Pathogenesis
The detailed mechanisms by which Actinomyces israelii causes
lumpy jaw have not yet been established. A. israelii cannot pen-
etrate the normal mucosal surface, but it can establish an infection
in association with other members of the normal microbiota if intro-
duced into tissue by wounds. The infectious process is characterized
0.25 mm
by alternate cycles of abscess formation and healing. The disease
usually progresses to the skin, where pus is discharged, but occa- FIGURE 23.12 Actinomyces israelii Sulfur Granule
sionally it penetrates into bone or into the central nervous system. Microscopic view of a stained smear of pus from an infected person.
In the tissue, A.  israelii grows as dense yellowish colonies Filaments of the bacteria can be seen radiating from the edge of
called sulfur granules because they are the color and size of par- the colony.
ticles of sulfur (figure  23.12). Finding these sulfur granules in ? What are the sulfur granules in the pus?
 Part IV Infectious Diseases 561

the wound drainage helps significantly with diagnosis. Typically, MicroAssessment 23.3
diagnosis depends on culturing the organism from the pus dis-
charge, but because A. israelii is slow growing, other more rapidly Tetanus (lockjaw) is caused by an anaerobic, spore-forming bacterium
with little invasive ability. The organisms release an exotoxin
growing bacteria present in the pus may outcompete it in culture, called tetanospasmin that is carried to the nervous system, causing
confusing the diagnosis. The presence of sulfur granules in the uncontrolled muscle spasms and possibly death. Gas gangrene
wound discharge confirms A. israelii infection. (clostridial myonecrosis), caused by another anaerobic sporeformer,
usually starts in neglected wounds containing dead tissue and
Epidemiology foreign material and spreads into normal muscle tissue. Lumpy jaw
(actinomycosis) is caused by a branching, filamentous, slow-growing,
Actinomyces israelii can be part of the normal microbiota of the
anaerobic bacterium, not by a fungus as the name implies. The disease
mucosal surfaces of the mouth, upper respiratory tract, intestine, is characterized by recurrent abscesses that drain characteristic sulfur
and sometimes the vagina. It is common in the gingival crevice, granules.
particularly in people with poor dental care. Pelvic actinomycosis
7. Why do many tetanus victims fail to respond to tetanus antitoxin?
can complicate use of intrauterine contraceptive devices (IUDs).
8. What factors lead to the development of gas gangrene?
The species of Actinomyces responsible for actinomycosis of
dogs, cattle, sheep, pigs, and other animals generally do not cause 9. Do babies need to be immunized against tetanus if their mother
had been immunized against the disease? Explain. +
human disease. The disease is sporadic and is not transmitted from
person to person. gingival crevice, p. 573

Treatment and Prevention

Actinomycosis can be treated with a number of antibacterial
23.4 ■ Bacterial Infections
medications, including penicillin and tetracycline. To be effective, of Bite Wounds
however, treatment must be given over weeks or months because
the organisms grow very slowly, and in dense colonies, into which Learning Outcomes
medications do not easily diffuse. 7. Explain why human mouth microbiota can cause serious bite
There are no proven preventive measures for lumpy jaw. The wound infections.
main features of actinomycosis are presented in table 23.5. 8. Describe two zoonotic wound infections.

Each year, more than 3 million animal bites occur in the United
States, the most feared result being the viral disease, rabies.
TABLE 23.5 Actinomycosis (“Lumpy Jaw”) Human bites are also common. The infection risk following a bite
Signs and Chronic disease; recurrent, sometimes
depends partly on the type of injury (crushing, lacerated, or punc-
symptoms painful swellings open and drain pus, heal ture), and partly on the kinds of pathogens introduced into the bite
with scarring; usually involves face and wound. rabies, p. 658
neck; chest, abdomen, and pelvis are other
common sites
Causative agent Actinomyces israelii, a filamentous, Human Bites
branching, Gram-positive, slow-growing,
Wounds caused by human teeth (such as bites that break the skin,
anaerobic bacterium
hitting the teeth of another person, or being injured with objects
Incubation Months (usually indeterminate)
that have been in a person’s mouth) are common and can result
period
in very serious infections. These infections are caused by nor-
Pathogenesis Usually begins in a mouth wound,
mal mouth microbiota. Occasionally, diseases such as syphilis,
extends to the face, neck, or upper chest;
sometimes begins in the lung, intestine, hepatitis B, and hepatitis C are transmitted this way, because the
or female pelvis. A. israelii always causative agents of these diseases are found in the blood or saliva
accompanied by normal microbiota. In (see Perspective 23.1).
tissue, grows as dense yellowish colonies
called sulfur granules.
Signs and Symptoms
Epidemiology No person-to-person spread. A. israelii
The wound may appear minor at first but then becomes painful
commonly part of normal mouth,
upper respiratory, intestine, and vagina and swells massively. Pus that smells very bad often leaks out of
microbiota. Dental procedures, intestinal the wound.
surgery, insertion of IUDs can initiate
infections. Causative Agents
Treatment Because of its slow growth, A. israelii The most frequent causes of infection in human bite wounds are
and prevention infections require prolonged treatment; a
both aerobic and anaerobic members of the normal mouth micro-
number of antibacterial medications are
effective. No proven preventive measures. biota, including streptococci, fusiforms, spirochetes, and Bacteroides
species, often in association with Staphylococcus aureus.
562 Chapter 23 Wound Infections

PERSPECTIVE 23.1
Infection Caused by a Human “Bite”
A 26-year-old man injured a finger of his cut open the infected tissues, allowing pus to was given antibiotics to fight infection, but the
right hand during a bar fight when he punched discharge. He removed the damaged tissue and wound did not heal well and continued to drain
someone in the mouth. The man did not seek washed the wound with sterile fluid. Smears pus. Several weeks later, X rays revealed that
medical help until more than 36 hours after and cultures of the infected material showed infection had spread to the bone at the base of
the fight. At that time, his entire hand was aerobic and anaerobic bacteria characteristic the finger. To cure the infection, the finger had
very swollen, red, and tender, and the swell- of mouth normal microbiota, including species to be amputated.
ing was spreading to his arm. The surgeon of Bacteroides and Streptococcus. The patient

Pathogenesis Pasteurella multocida Bite

Bite wounds are usually infected with both aerobic and anaerobic Wound Infections
species. Crushed tissue and the presence of facultative anaer-
Infections caused by bacteria that live in the mouth of a biting ani-
obes, which reduce available O2, create anaerobic conditions
mal are very common. Surprisingly, bite infections from a number
that allow anaerobic bacteria to establish infection at the site.
of kinds of animals are generally caused by a single bacterial spe-
Although most mouth microbiota are harmless alone, together
cies, Pasteurella multocida.
they produce large numbers of toxins and enzymes that damage
the host tissues and cells involved in the immune response. These Signs and Symptoms
include leukocidin, collagenase, hyaluronidase, ribonuclease, Early signs of infection in wounds caused by animal bites include
various proteinases, neuraminidase, and enzymes that destroy spreading redness, tenderness and swelling of tissues next to the
antibodies and proteins of the complement system. The result wound, followed by pus discharge. Abscesses commonly form.
of all these factors is a synergistic infection, meaning that the Without quick treatment, infection can lead to bloodstream inva-
sum effect of all the organisms acting together is greater than the sion or permanent loss of function.
sum of their individual effects. Irreversible destruction of tissues
such as tendons and permanent loss of function can be the result. Causative Agent
facultative anaerobes, p. 90 Pasteurella multocida is a Gram-negative, facultatively anaerobic
coccobacillus. Most isolates have antiphagocytic capsules. There
Epidemiology are a number of different antigenic types.
Most serious human bite infections occur because of fights
related to drunkenness, or during forcible restraint, as found in Pathogenesis
law enforcement and in mental institutions. The risk is greatly The details of pathogenesis are not yet known. Some strains pro-
increased when the biting individual has poor mouth care and duce a toxin that kills host cells by affecting protein synthesis and
extensive dental disease. Bites by young children are usually not cell cycle regulation. The organisms have antiphagocytic capsules
serious, mostly because children seldom break the skin or cause
crushing wounds when they bite.
TABLE 23.6 Human Bite Wound Infections
Treatment and Prevention Signs and Rapid onset, pain, massive swelling, drainage
Bite wounds should be treated immediately to avoid serious infec- symptoms of foul-smelling pus
tions. This involves opening the infected area with a scalpel, wash- Incubation Usually 6 to 24 hours
ing the wound thoroughly with sterile fluid, and removing dirt, period
foreign matter (such as broken teeth), and dead tissue. Antibacterial Causative agent Mixed mouth microbiota: anaerobic
medications should also be given. Usually more than one drug is streptococci, fusiforms, spirochetes,
used because many oral microbiota involved in bite wound infec- anaerobic Gram-negative rods; sometimes
Staphylococcus aureus
tions are resistant to various drugs such as penicillin. One drug
effective against anaerobes should be included. If bite wounds are Pathogenesis Various mouth bacteria act synergistically to
destroy tissue.
not treated quickly, serious complications may occur, including
infection of muscles and tendons, which may require surgery. Epidemiology Alcohol-related violence; forcible restraint;
Prevention of bite wound infections involves avoiding situa- poor mouth care and extensive dental
disease.
tions that lead to biting and hitting. If a bite wound is sustained,
prompt cleaning of the wound followed by application of an anti- Treatment Treatment is usually surgical. No proven
and prevention preventive measures except to avoid fights.
septic can help prevent infection. The main features of human bite Prompt cleaning of wound and application
wound infections are presented in table 23.6. of antiseptic is advised.
 Part IV Infectious Diseases 563

but when antibodies bind to them, phagocytes are able to ingest

and kill the opsonized bacteria. If untreated, P. multocida can
cause bacteremia, leading to endocarditis or meningitis.

MicroByte
Using Pasteurella multocida, Pasteur demonstrated that attenuated
organisms could be used as a vaccine.

Epidemiology
Humans—and many other healthy animals such as cats, dogs, and
monkeys—carry P.  multocida as normal oral and upper respira-
tory microbiota. Both diseased and healthy animals are reservoirs
for human infections. Cats are more likely to carry P. multocida
than dogs, and so cat bites more frequently cause the infection.
P. multocida also causes diseases in a number of other animal
FIGURE 23.13 Bartonellosis (“Cat Scratch Disease”) Note the
species, sometimes causing epidemics of fatal pneumonia and wrist lesion and enlarged lymph nodes
bloodstream infection in them.
? Is this disease only transmitted by cat scratches?
Treatment and Prevention
Unlike many Gram-negative pathogens, P. multocida is suscepti-
ble to penicillin, so amoxicillin clavulanate (Augmentin) is given
Bartonellosis (“Cat Scratch Disease”)
immediately, even before the diagnosis has been confirmed by
culturing the bacterium. This drug is a combination of a penicillin In the United States, bartonellosis (commonly known as “cat
derivative (amoxicillin) and a β-lactamase inhibitor (clavulanate). scratch disease”) is the most common cause of chronic lymph
The clavulanate is helpful because many bite wounds often also node enlargement at one body site in young children. Despite its
contain strains of β-lactamase-producing Staphylococcus aureus. name, this disease can also be transmitted by bites and possibly
This and other antibacterial medications are effective if given by other means such as fleabites or saliva on mucous membranes.
early in the infection. Typically the infection is mild and localized, lasting from several
No vaccines for P. multocida are available for use in humans. weeks to a few months.
Immediate cleaning of bite wounds and quick medical attention
usually prevent the development of serious infection. Table 23.7 Signs and Symptoms
gives the main features of Pasteurella multocida bite wound Cat scratch disease begins within a week of a scratch or bite with
infections. the appearance of a pus-filled pimple at the site of injury. Painful
enlargement of the lymph nodes of the area develops in 1 to 7
weeks (figure  23.13). Patients may develop fever, and in some
Pasteurella multocida Bite cases the lymph nodes become pus-filled and soft. The disease
TABLE 23.7 generally disappears without treatment in 2 to 4 months. A small
Wound Infections
number of patients develop irritation of an eye with local lymph
Signs and Spreading redness, tenderness, swelling, node enlargement, epileptic seizures and coma due to encephali-
symptoms discharge of pus
tis, or acute or chronic fever associated with bloodstream or heart
Incubation 24 hours or less valve infection.
period
Causative Pasteurella multocida, a Gram-negative, Causative Agent
agent facultatively anaerobic, encapsulated
coccobacillus
Cat scratch disease is caused by Bartonella henselae, a curved,
Gram-negative rod.
Pathogenesis Introduced by bite, P. multocida attaches
to tissue, resisting phagocytes because of
its capsule; probable cell-destroying toxin. Pathogenesis
Extensive swelling, abscess formation. The virulence factors of B. henselae and the process by which
Opsonins develop, allow phagocytic killing, it causes disease are not yet known. The organisms can enter
limit spread.
the body by a cat bite or scratch, or when cat saliva con-
Epidemiology Carried by many animals in their mouth or taminates a mucous membrane. The bacteria are carried to
upper respiratory tract.
the lymph nodes, and the disease is resolved by the immune
Treatment Treatment with penicillin, other system in most cases. Spreading by the bloodstream, however,
and prevention antibacterials, effective if given promptly.
No vaccines to use in humans. Prompt wound
occurs in some individuals, causing serious complications.
care is preventive. Peliosis hepatis and bacillary angiomatosis are two complicat-
ing conditions seen mostly in people with AIDS. In peliosis
564 Chapter 23 Wound Infections

hepatis, blood-filled cysts form in the liver. In bacillary angi- Streptobacillary Rat Bite Fever
omatosis, nodules composed of proliferating blood vessels
Rat bites are fairly common among poor people in large cities and
develop in the skin and other parts of the body.
among workers who handle laboratory rats.
Epidemiology
Signs and Symptoms
Cat scratch disease occurs worldwide, often in children and
The bite wound usually heals quickly without any problems. Two
young adults. The disease is a zoonosis, transmitted to humans
to 10 days later, however, the person develops chills, fever, head
mainly by bites or scratches from cats. Person-to-person spread
and muscle aches, and vomiting. The fever comes and goes. A
does not occur. Asymptomatic bacteremia is common in cats,
rash usually appears after a few days, followed by joint pain.
however, and transmission from cat to cat occurs by cat fleas.
Fleas probably are responsible for transmission in cases where
Causative Agent
people develop the disease after they handle cats but are not
scratched or bitten. The cause of streptobacillary rat bite fever is Streptobacillus
moniliformis, a facultatively anaerobic, Gram-negative rod that
Treatment and Prevention varies in shape (pleomorphic). The organism is unique in that it
spontaneously develops L-forms. L-forms are variants that lack a
Any cat scratch or bite should be promptly cleaned with soap
cell wall, first identified at the famous Lister Institute (hence, the L
and water and then treated with an antiseptic. If signs of infec-
in L-form). As might be expected, L-form colonies resemble those
tion develop, or if the cat’s immunization status against rabies
of mycoplasmas—bacteria that lack a cell wall. mycoplasmas, p. 276
is uncertain, prompt medical evaluation is needed. Severe
B. henselae infections can usually be treated with antibacterial
Pathogenesis
medications such as ampicillin, but some strains are resistant to
this drug. Streptobacillus moniliformis enters the body through a bite or
There are no proven preventive measures for cat scratch scratch, and sometimes by ingestion. There is typically little
disease, other than to avoid handling stray cats, especially those enlargement of the local lymph nodes, and the bacteria quickly
with fleas. The main features of cat scratch disease are presented enter the bloodstream and spread throughout the body. The major-
in table 23.8. ity of people recover without treatment in about 2 weeks, but some
cases are rapidly fatal, and others develop serious complications
such as brain abscesses or infection of the heart valves. The main
features of streptobacillary rat bite fever are presented in table 23.9.
TABLE 23.8 Bartonellosis (“Cat Scratch Disease”)
Signs and Pimple appears at the bite or scratch site, TABLE 23.9 Streptobacillary Rat Bite Fever
symptoms followed by local lymph node enlargement;
nodes may soften and drain pus; prolonged Signs and Chills, fever, muscle aches, headache, and
fever and convulsions indicate spread to symptoms vomiting; later, rash and pain in one or
other body parts more of the large joints
Incubation Usually less than 1 week Incubation Usually 2 to 10 days (range, 1 to 22) after a
period period rat bite
Causative Bartonella henselae, a Gram-negative rod Causative Streptobacillus moniliformis, a highly
agent agent pleomorphic, Gram-negative bacterium
that spontaneously produces L-forms
Pathogenesis Bacteria enter with cat bite or scratch
and reach lymph nodes, where the Pathogenesis Bite wound heals without treatment;
disease is usually stopped. May spread by S. moniliformis quickly invades
bloodstream, cause infections of the heart, bloodstream. Fevers come and go
brain, or other organs. Peliosis hepatis or irregularly. Most victims recover without
bacillary angiomatosis may occur, mostly in treatment; in others, infection established
people with AIDS. in various body organs, results in death if
treatment is not given.
Epidemiology A zoonosis, spread among cats by fleas,
which are biological vectors and have Epidemiology Wild and laboratory rats can carry
B. henselae in their feces. Infected cats S. moniliformis. Bites of other rodents and
usually asymptomatic, but often bacteremic. animals that prey on them can transmit
Humans are accidental hosts. No person- the disease to humans. Food or drink
to-person spread. contaminated with rodent feces can also
transmit the infection.
Treatment Promptly wash skin breaks, apply
and prevention antiseptic. Most B. henselae infections Treatment Effectively treated with penicillin, other
respond to antibacterial treatment. and prevention antibacterial medications. Control wild
Avoiding rough play with cats. Flea control rats and mice; care in handling laboratory
in cats. animals.
 Part IV Infectious Diseases 565

Epidemiology
Many healthy laboratory rats, mice, and other rodents carry
S. moniliformis in their nose and throat. Laboratory and pet store
workers are at increased risk of contracting the disease. Rat bites
and scratches are the usual source of human infections. Epidemics
of the disease have occurred when people have eaten or drunk
anything contaminated with S.  moniliformis from rodent feces.
Cases of rat bite fever have also been associated with exposure to
animals that eat rodents, including cats and dogs.

FIGURE 23.14 Individual with Sporotrichosis

Treatment and Prevention (“Rose Gardener’s Disease”)
Penicillin, given intravenously, is used to treat cases of rat ? Why do the abscesses occur along a relatively straight line?
bite fever. Since this drug inhibits cell wall synthesis, the fact
that it can be used to treat rat bite fever shows that either
S.  moniliformis L-forms do not occur in vivo, or if they do,
they are avirulent. Prevention of rat bite fever includes control
Sporotrichosis
of wild rat populations and care in handling laboratory rats and
their feces. (“Rose Gardener’s Disease”)
Sporotrichosis, also known as “rose gardener’s disease,” occurs
around the world and is associated with activities that lead to
MicroAssessment 23.4 puncture wounds from plant material. Although many cases are
Human bite infections can be dangerous because certain members of sporadic, the disease can occur in groups of people working in
the mouth microbiota, which have little invasive ability when growing the same occupation, such as farmers, gardeners, and agricultural
alone, can invade and destroy tissue when growing synergistically. workers. Veterinarians are also at risk from animals such as cats
Pasteurella multocida can infect bite wounds caused by a number that have contracted the disease.
of different animals, especially cats. Cat bites and scratches can also
transmit Bartonella henselae, the cause of cat scratch disease, which
is characterized typically by local lymph node enlargement, although Signs and Symptoms
it may involve other parts of the body. Streptobacillary rat bite fever, The site of infection in most cases is a hand or arm but the body,
caused by Streptobacillus moniliformis, is acquired from bites of legs, and face can also be involved. Typically, a chronic ulcer
rats and mice, and from animals that eat them. It is characterized by
forms at the wound site, followed by a slowly progressing series
fever that comes and goes and a rash. S. moniliformis spontaneously
develops L-forms. of ulcerating nodules that develop sequentially toward the center
of the body (figure  23.14). Lymph nodes in the region of the
10. What Gram-negative organism commonly infects wounds caused
wound enlarge, but patients generally do not become ill. If they
by animal bites?
have AIDS or other immunodeficiency, however, the disease can
11. What is the most common cause of chronic localized lymph node
spread throughout the body, infecting joints and the central nerv-
enlargement in young children?
ous system, and becoming life-threatening.
12. Why are normal mouth microbiota a cause of serious infection in
human bites? +
Causative Agent
Sporotrichosis is caused by the dimorphic fungus Sporothrix
schenckii, which lives in soil and on vegetation (figure  23.15).
dimorphic fungus, p. 285
23.5 ■ Fungal Wound
Infections Pathogenesis
Sporothrix schenckii spores enter the body through an injury
Learning Outcome caused by plant material. After an incubation period that usually
9. Give the distinctive features of rose gardener’s disease. ranges from 1 to 3 weeks but can be longer, the multiplying fungi
cause a small pimple to form at the site of the injury. This slowly
Fungal infections of wounds are more serious than dermal mycoses. enlarges and ulcerates, producing a painless, red lesion that bleeds
Typically, they are caused by soil fungi that enter through injuries easily. There is little or no pus unless the ulcer becomes second-
that allow them to invade subcutaneous tissue. Sporotrichosis is arily infected with bacteria. After a week or longer, the process
a common fungal infection that occurs worldwide. It is typically repeats itself—progression of the disease usually follows the flow
a mild disease but may become life-threatening under certain of a lymphatic vessel. In healthy individuals, the process does not
circumstances. proceed beyond the lymph node. Sometimes, however, satellite
566 Chapter 23 Wound Infections

Sporotrichosis (“Rose
TABLE 23.10 Gardener’s Disease”)
Spores

Signs and Painless, ulcerating nodules appearing in

symptoms sequence in a linear pattern

Hyphae Incubation period Usually 1 to 3 weeks

Causative agent Sporothrix schenckii, a dimorphic fungus
Pathogenesis Spores multiply at site of introduction by
thorn, splinter, or other plant material,
causing a small nodule that ulcerates.
(a) 25 μm Spores carried by lymph flow repeat the
process along the lymphatic vessel. May
spread beneath the skin irrespective of
lymphatic vessels.
Epidemiology Distributed worldwide in tropical and
temperate climates. Occupations requiring
Yeast forms contact with sharp plant materials at
particular risk.
Treatment Most cases effectively treated with
and prevention potassium iodide. Generalized infections
require amphotericin B or itraconazole.
Protective gloves and clothing.

(b) 30 μm

FIGURE 23.15 Sporothrix schenckii (a) Mold form. (b) Yeast

form, as seen in infected tissue.
? Sporothrix schenckii is dimorphic; what does this mean? ability to fight the fungus. Itraconazole or the antifungal medica-
tion amphotericin B is used in rare cases when the disease spreads
throughout the body. Sporotrichosis is often misdiagnosed, lead-
ing to delayed and inappropriate treatment.
lesions appear irrespective of the lymphatic vessels. Without Sporotrichosis can be prevented by wearing protective gloves
treatment, the disease can become chronic and go on for years. and a long-sleeved shirt when working with soil and vegetation.
lymphatic vessel, p. 358 Table 23.10 presents some of the main features of sporotrichosis.

Epidemiology The key features of the diseases covered in this chapter are high-
Sporotrichosis is distributed worldwide, mostly in the warmer lighted in the Diseases in Review 23.1 table.
regions but extending into temperate climates. It is an occupa-
tional disease of farmers, carpenters, gardeners, greenhouse work- MicroAssessment 23.5
ers, and others who work with plant materials. Risk factors for the
disease besides occupation include diabetes, immunosuppression, Rose gardener’s disease (sporotrichosis), caused by the dimorphic
fungus Sporothrix schenckii, is widely distributed around the world,
and alcoholism. Individuals with chronic lung disease can contract affecting mainly those associated with occupations that expose
S. schenckii lung infections from inhaling dust from hay or cattle them to splinters and sharp vegetation. Unlike most invasive fungal
feed. Deaths from sporotrichosis are rare. infections, sporotrichosis is usually easy to treat.
13. List some of the risk factors associated with contracting rose
Treatment and Prevention gardener’s disease.
Surprisingly, unlike other fungal infections, sporotrichosis can 14. What is unique about the treatment of sporotrichosis?
usually be cured by oral treatment with the simple chemical com- 15. Why is sporotrichosis often misdiagnosed? +
pound potassium iodide (KI). KI somehow enhances the body’s
Diseases in Review 23.1
Wound Infections

Disease Causative Agent Comment Summary Table

BACTERIAL INFECTIONS

Staphylococcal Staphylococcus aureus S. aureus (described in chapter 22) is the most common cause Table 23.1, p. 551
infections and S. epidermidis of wound infections, but S. epidermidis, part of the normal skin
microbiota, can form biofilms on medical devices.
Necrotizing fasciitis SPE-producing strains of SPEs of S. pyogenes (described in chapter 21) are superantigens, Table 23.1, p. 551
(“flesh-eating disease”) Streptococcus pyogenes which cause widespread and inappropriate activation of helper
T cells.
Pseudomonas Pseudomonas aeruginosa Widespread environmental organism that commonly infects Table 23.1, p. 551
aeruginosa infections burned tissues; produces a green pigment; resistant to a wide
variety of antimicrobial medications.

DISEASES DUE TO ANAEROBIC BACTERIA

Tetanus (“lockjaw”) Clostridium tetani Endospore-forming anaerobe grows in wound-damaged tissues; Tables 23.2 and
tetanus toxin blocks the action of inhibitory neurons, leading to 23.3, p. 557
continuous painful muscle contractions; typically fatal without
treatment. Preventable by vaccination (DTaP).
Clostridial myonecrosis Usually Clostridium Endospore-forming anaerobe that can germinate and grow Table 23.4, p. 559
(“gas gangrene”) perfringens in necrotic and poorly oxygenated tissue; gases produced
accumulate in tissues; typically fatal without treatment.
Actinomycosis (“lumpy Usually Actinomyces israelii Characterized by recurring and slowly progressing swellings, Table 23.5, p. 561
jaw”) usually near the neck and jaw; these burst, draining pus and
sulfur granules.

BITE WOUND INFECTIONS

Human bites Mixed aerobic and anaerobic Crushing nature of wound causes tissue damage and anaerobic Table 23.6, p. 562
species, often members of the conditions; synergistic infection adds to that tissue damage.
normal mouth microbiota
Pasteurella multocida Pasteurella multocida Characterized by abscess at the site of the wound; causative Table 23.7, p. 563
infections agent is a common member of the normal mouth microbiota of
animals; results most often from cat bites.
Bartonellosis (“cat Bartonella henselae Most common cause of lymph node enlargement at one site in Table 23.8, p. 564
scratch disease”) children; acquired via a cat bite or scratch.
Streptobacillary rat Streptobacillus moniliformis Characterized by relapsing fevers, rash, and joint pain. Table 23.9, p. 564
bite fever Acquired via a rat bite or scratch, or ingestion of food or water
contaminated with rodent feces.

FUNGAL WOUND INFECTIONS

Sporotrichosis (“rose Sporothrix schenckii Characterized by painless ulcerating nodules that develop Table 23.10, p. 566
gardener’s disease”) sequentially along a lymphatic vessel; dimorphic fungus lives on
soil and vegetation and enters the body through an injury.
568 Chapter 23 Wound Infections

FUTURE CHALLENGES 23.1

Staying Ahead in the Race with Staphylococcus aureus
Over the last 50 years, scientists have gener- understanding staphylococcal pathogenesis. reduce staphylococcal infections in kidney
ally kept us at least one step ahead in the race We now understand the mechanism by which dialysis patients.
between Staphylococcus aureus and human S.  aureus obtains iron from hemoglobin. We Other options now seem to be within
health. Today, “staph” seems again ready to also know that the bacterium’s protein A can reach. For example, it should be possible to
cause problems, but new knowledge from attach to tumor necrosis factor (TNF) recep- determine the structure of the active part of
molecular biology promises to help us fight tors on host cells, causing release of cytokines toxin molecules and to design medications
this pathogen. that recruit large numbers of neutrophils. that would bind to that site and inactivate it.
In the past, we have relied on developing There have also been advances in vaccine Also, it is now possible to determine which
antibacterial medications to kill or inhibit development. An experimental vaccine com- staphylococcal genes code for virulence fac-
the growth of S. aureus, but there is continu- posed of staphylococcal capsule material con- tors. The products of these genes may prove to
ing investigation of alternative approaches. jugated with a non-toxic form of Pseudomonas be unexpected vaccine candidates.
This search is aided by improvement in exotoxin A has been shown to significantly The race continues!

Summary
23.1 ■ Anatomy, Physiology, and Ecology Pseudomonas aeruginosa Infections
Wounds expose tissue components to which pathogens specifically Pseudomonas aeruginosa, an aerobic, Gram-negative rod with a single
attach. Wounds heal by forming granulation tissue which, in the polar flagellum, is an opportunistic pathogen widespread in the environ-
absence of dirt and infection, fills the defect, and subsequently contracts ment, and a cause of both healthcare-associated infections and those
to minimize scar tissue (figure 23.1). Thermal burns often result in large acquired outside the hospital. Production of two pigments by the bacte-
areas of dead tissue lacking competing organisms and body defenses, rium often colors infected wounds green (figure 23.6).
ideal conditions for microbial growth.
Wound Abscesses 23.3 ■ Diseases Due to Anaerobic Bacterial
An abscess is composed of pus, which contains leukocytes, tissue Wound Infections
breakdown products, and infecting organisms. Abscess formation local- Tetanus (“Lockjaw”) (table 23.3)
izes an infection within tissue, preventing its spread. Inflammatory cells
The characteristic symptom of tetanus is sustained, painful, cramp-
and clotted blood vessels separate the abscesses from normal tissue
like spasms of one or more muscles (figure  23.7). The disease is often
(figure 23.2).
fatal. Tetanus is caused by an exotoxin, tetanospasmin, produced
Anaerobic Wounds by Clostridium tetani, a non-invasive, anaerobic, Gram-positive rod (fig-
Anaerobic conditions are likely to occur in wounds containing dead tis- ure 23.8). The toxin prevents inhibitory neurons from releasing their neu-
sue or foreign material, and those with limited opening to the air. rotransmitter, causing muscles to contract without control (figure 23.9).
The spores of C. tetani are widespread in dust and dirt. Tetanus can be
23.2 ■ Common Bacterial Infections of Wounds (table 23.1) prevented by vaccination with toxoid (inactivated tetanospasmin), and
Possible consequences of wound infections include delayed healing, maintaining immunity throughout life with regular booster injections.
abscess formation, and spread of bacteria or toxins into nearby tissue or People sustaining any wound, including surgeries, no matter how minor,
the bloodstream. Infections can cause surgical wounds to open, and they should make sure that their immunization is up to date (table 23.2).
can spread to create biofilms on artificial devices.
Clostridial Myonecrosis (“Gas Gangrene”) (table 23.4)
Staphylococcal Wound Infections (figures 23.3, 23.4) Usually caused by the α-toxin-producing anaerobe Clostridium
Staphylococci are the leading cause of wound infections, both surgical perfringens. Signs and symptoms begin suddenly with pain and swell-
and accidental; Staphylococcus aureus and S. epidermidis are the most ing followed by discharge of a thin, brown, bubbly fluid and dark
common wound-infecting species. Staphylococcus aureus possesses discoloration of the overlying skin (figure 23.10). The toxin causes tissue
many virulence factors; some strains release a toxin that causes toxic necrosis; hydrogen and carbon dioxide are produced from fermentation
shock syndrome (table 23.1). Staphylococcus epidermidis is less virulent of amino acids and glycogen in the dead tissue. There is no vaccine or
but can form biofilms on catheters and other devices. toxoid. Prevention depends on prompt medical care of dirty wounds
containing dead tissue. Treatment depends on urgent surgical removal
Group A Streptococcal “Flesh-Eating Disease”
of dead and infected tissue, and it may require amputation.
Streptococcus pyogenes (group A, β-hemolytic streptococcus) causes
strep throat, scarlet fever, wound infections, and other conditions. Actinomycosis (“Lumpy Jaw”) (table 23.5)
Necrotizing fasciitis–causing strains of S.  pyogenes produce exo- Actinomycosis is a chronic, slowly progressive disease characterized
toxin B, a protease thought to be responsible for the tissue destruction by repeated swellings, discharge of pus, and scarring, usually of the
(figure 23.5). face and neck (figure 23.11). The causative agent is Actinomyces israelii,
 Part IV Infectious Diseases 569

a member of the normal microbiota of the mouth, intestine, and vagina it begins with a pimple at the site of a bite or scratch, followed by
that enters tissues with wounds such as those with dental and intestinal enlargement of local lymph nodes, which often become pus-filled. Most
surgery (figure 23.12). Despite its name, it is not a fungus. The organism individuals with cat scratch disease recover without treatment.
is slow growing; treatment must be continued for weeks or months.
Streptobacillary Rat Bite Fever (table 23.9)
23.4 ■ Bacterial Infections of Bite Wounds Streptobacillary rat bite fever is characterized by relapsing fevers, head
The kind of bite wound infection depends on the kinds of infectious and muscle aches, and vomiting. A rash and joint pains often develop.
agents in the mouth of the biting animal, and the nature of the wound— It is usually caused by Streptobacillus moniliformis, a pleomorphic,
whether punctured, crushed, or torn. Gram-negative rod that characteristically produces cell wall–deficient
variants called L-forms. People who live in rat-infested areas and those
Human Bites (table 23.6)
who handle laboratory rats are at greatest risk of contracting the disease.
Wounds caused by human teeth are common and can result in very seri-
ous infections, with pain, massive swelling, and foul-smelling pus. The
23.5 ■ Fungal Wound Infections
infections are usually caused by synergistic activity among members of the
Fungal wound infections are usually more serious than dermal mycoses.
normal mouth microbiota, including anaerobic streptococci, fusiforms, spi-
They are typically caused by fungi from soil.
rochetes, and Bacteroides sp., often with Staphylococcus aureus. The crush-
ing nature of bite wounds causes death of tissue and conditions suitable for Sporotrichosis (“Rose Gardener’s Disease”) (table 23.10)
growth of anaerobes; prompt cleansing and use of an antiseptic are advised.
Rose gardener’s disease, also called sporotrichosis, is a chronic fungal
Pasteurella multocida Bite Wound Infections (table 23.7) disease mainly of people who work with soil or vegetation. The usual
Pasteurella multocida, a Gram-negative rod, can infect bite wounds case is characterized by painless, ulcerating nodules that develop one
inflicted by a number of animal species. P. multocida causes disease in after the other along the path of a lymphatic vessel (figure  23.14). The
animals, but many animals are asymptomatic carriers. causative organism is the dimorphic fungus, Sporothrix schenckii
(figure  23.15), usually introduced into wounds caused by thorns or
Bartonellosis (“Cat Scratch Disease”) (table 23.8, figure 23.13) splinters. The fungus is distributed worldwide in tropical and temper-
Cat scratch disease is the most common cause of chronic, localized ate climates; people at risk include farmers, carpenters, gardeners, and
lymph node enlargement in children. Caused by Bartonella henselae, greenhouse workers.

Review Questions
Short Answer 2. Which of these statements about Streptococcus pyogenes is false?
1. What property of Staphylococcus epidermidis help it to colonize a) It is a Gram-positive coccus occurring in chains.
plastic materials used in medical procedures? b) Some strains that infect wounds can cause toxic shock.
2. What is the relationship between the superantigens of S.  aureus c) Some strains that infect wounds can cause necrotizing fasciitis.
and the organism’s production of toxic shock? d) It can cause puerperal sepsis.
3. Name two underlying conditions that predispose a person to e) A vaccine is available for preventing S. pyogenes infections.
Streptococcus pyogenes flesh-eating disease. 3. Choose the one false statement about Pseudomonas aeruginosa.
4. Give two sources of Pseudomonas aeruginosa. a) It is widespread in nature.
5. Outline the pathogenesis of tetanus. b) Some strains can grow in distilled water.
6. Explain why C. tetani can be cultivated from wounds in the c) It is a Gram-positive rod.
absence of tetanus. d) It produces a hemolytic toxin.
7. What characteristics of bite wounds lead to anaerobic infections? e) Under certain circumstances, it can grow anaerobically.
8. What is the causative agent of cat scratch disease? Why is it a 4. Which of these statements about tetanus is true?
threat to patients with AIDS? a) It can start from a bee sting.
9. What is a synergistic infection? How might one be acquired? b) Immunization is carried out using tiny doses of killed C. tetani.
10. Why is sporotrichosis sometimes called rose gardener’s disease? c) Those who recover from the disease are immune for life.
d) Tetanus immune globulin does not prevent the disease.
Multiple Choice e) It is easy to avoid exposure to spores of the causative organism.
1. Which of the following about Staphylococcus aureus is false? 5. Choose the one true statement about gas gangrene.
a) It is generally coagulase-positive. a) There are few or no leukocytes in the wound drainage.
b) Its infectious dose is increased in the presence of foreign material. b) It is best to rely on antibacterial medications and avoid
c) Some strains infecting wounds can cause toxic shock. disfiguring surgery.
d) Nasal carriers have an increased the risk of surgical wound infection. c) A toxoid is generally used to protect against the disease.
e) It is pyogenic. d) Only one antitoxin is used for treating all cases of the disease.
e) It is easy to avoid spores of the causative agent.
570 Chapter 23 Wound Infections

6. Which of the following statements about actinomycosis is false? 10. Which statement concerning sporotrichosis is false?
a) It can occur in cattle. a) It is characterized by ulcerating lesions along the path of a
b) It is caused by a branching filamentous bacterium. lymphatic vessel.
c) It always appears on the jaw. b) Person-to-person transmission is common.
d) It can arise from intestinal surgery. c) It can occur in epidemics.
e) Its abscesses can penetrate bone. d) It can persist for years if not treated.
7. Which of the following statements about Pasteurella multocida e) The causative organism is a dimorphic fungus.
is false?
Applications
a) Infections generally respond to a penicillin.
1. Clinicians become concerned when the laboratory reports that
b) It can cause epidemics of fatal disease in domestic animals.
organisms capable of digesting collagen and fibronectin are pres-
c) It is commonly found in the mouths of biting animals, including ent in a wound culture. What is the basis of their concern?
humans.
2. An army field nurse working at a mobile surgical hospital asks this
d) A vaccine is used to prevent P. multocida disease in people.
question of all the ambulance drivers: “Was the soldier wounded
e) Cat bites are more likely to result in P. multocida infections while in a field with cows?” Why does the nurse ask this question?
than dog bites.
8. Which of these statements about cat scratch disease is false? Critical Thinking +
a) It is a common cause of chronic lymph node enlargement in 1. In what way would the incidence of tetanus at various ages in a devel-
children. oping country differ from age incidence in developed countries?
b) It is a serious threat to individuals with AIDS. 2. Could colonization of a wound by a non-invasive bacterium cause
c) Cat scratches are the only mode of transmission to humans. disease? Explain your answer.
d) It is a zoonosis of cats transmitted by fleas.
e) It can affect the brain or heart valves in a small percentage of cases.
9. The following statements about Streptobacillus moniliformis are
all true except
a) it can be transmitted by food.
b) its colonies can resemble those of mycoplasmas.
c) it can be transmitted by the bites of animals other than rats.
d) human infection is characterized by irregular fevers, rash, and
joint pain.
e) it is a Gram-positive spore-forming rod.
24 Digestive System Infections
KKE EYYT TE ERRMMS S
Antibiotic-Associated
Diarrhea Diarrhea that occurs as a
complication of taking antimicrobial
medications.
Gingivitis Inflammation of the
gums.
Hemolytic Uremic Syndrome
(HUS) Serious condition
Bile Fluid produced by the liver that characterized by red blood cell
aids in the digestion and absorption breakdown and kidney failure.
of fats. Hepatitis Inflammation of the
Cariogenic Causes dental caries liver.
(tooth decay). Microvilli Tiny extensions from the
Cirrhosis Scarring of the liver that surfaces of cells such as those lining
interferes with normal liver function. the intestinal villi; they increase
surface area of the mucosa.
Dysentery A serious form of
diarrhea characterized by blood, pus, Oral Rehydration Therapy
and mucus in the feces. (ORT) A treatment used to replace
fluid and electrolytes lost due to
Gastroenteritis Acute
diarrheal disease.
inflammation of the stomach and
intestines; the syndrome of nausea, Villi Short fingerlike protrusions
vomiting, diarrhea, and abdominal from the intestinal mucosa that slow
pain. the movement of food and increase
the surface area of the intestinal
lumen.

Yellow color of eye and skin as seen in jaundice.

A Glimpse of History elsewhere. The number of new cases decreased. The outbreak had already
An army surgeon wrote a vivid description of cholera in 1832 when the begun subsiding before the handle was removed, however, so Snow’s
disease first appeared in the United States: explanation that cholera was a waterborne disease was not accepted by
The face was sunken . . . perfectly angular, and rendered most doctors and government officials. By 1866, however, it was evident
peculiarly ghastly by the complete removal of all the soft solids, that cholera did indeed appear in areas that lacked sanitation, just as Snow
in their places supplied by dark lead-colored lines. The hands had proposed. Public health agencies then played a major role in prevent-
and feet were bluish white, wrinkled as when long macerated in ing epidemic cholera. In 1883 Robert Koch, who proved the germ theory
cold water; the eyes had fallen to the bottom of their orbes, and of disease, isolated Vibrio cholerae, the bacterium that causes cholera.
envinced a glaring vitality, but without mobility, and the surface
of the body was cold.

I 
n the fall of 2008, the first reports of a cholera outbreak in
Cholera is a very old disease, thought to have originated in the Far Zimbabwe began trickling in to the World Health Organization.
East thousands of years ago. Sanskrit writings indicate it existed in India Volunteers from the medical relief organization Medecins Sans
many centuries before Christianity. With the increased shipping of goods Frontieres (Doctors Without Borders) rushed to set up medical
and the mobility of people during the nineteenth century, cholera spread facilities, while people from the World Health Organization, Red
from Asia to Europe and then to North America. The disease caused Cross, and local health agencies worked to provide filtered and
major epidemics in the nineteenth century.
chlorinated drinking water, construct latrines, and educate people
John Snow, a London physician, demonstrated that cholera was
transmitted by contaminated water. He observed that almost all people
about sanitary measures. Patients were treated with oral and intra-
who contracted the disease during an outbreak in 1854 got their water venous rehydration fluids. The lack of public health resources,
from a well on Broad Street, whereas neighbors who got their water political instability in Zimbabwe, and global economic crisis
elsewhere were unaffected. Even though the germ theory of disease had combined to amplify this outbreak. The resulting epidemic sick-
not yet been described, Snow was able to convince local authorities to ened almost 100,000 people and killed nearly 2,300 individuals
remove the pump handle so that people were forced to get their water in Zimbabwe and surrounding countries by its end in July 2009.

571
572 Chapter 24 Digestive System Infections

24.1 ■ Anatomy, Physiology, food into small particles, processing those even further, and then
absorbing the available nutrients. The waste material that remains
and Ecology is discharged as feces.
The digestive system encompasses two general components—
Learning Outcomes the digestive tract and the accessory organs (figure  24.1). The
1. Describe the functions of the main components of the upper and digestive tract is a hollow tube that extends from the mouth to
lower digestive tract. the anus. When referring only to the stomach and the intestines,
2. Identify the role of the liver and accessory organs in health and the term gastrointestinal tract is often used. The accessory organs,
disease. which include the salivary glands, liver, and pancreas, support the
3. Describe the significance of the normal intestinal microbiota. process of digestion by producing vital enzymes and other sub-
stances that help break down the food.
The main purpose of the digestive system is to convert the food we Like the respiratory system and skin, the digestive tract is
eat into a form that the body’s cells can use as a source of energy one of the body’s major boundaries with the environment. It is
and raw materials for growth. It does this by first breaking down distinct, however, in that the foods being digested by the host

Organ Function

Oral cavity Obtains and

processes food

Salivary Secrete saliva

Parotid salivary gland glands
Oral cavity containing Mumps
tongue and teeth Esophagus Transports food to
Dental caries stomach
Periodontal disease Salivary glands

Esophagus Stomach Stores food; mechanical

Esophagitis digestion; breaks down
some proteins

Pancreas Secretes digestive

Stomach enzymes
Liver Gastritis
Hepatitis Gastric ulcer
Liver Produces bile to assist
Gallbladder Pancreas in fat digestion
Pancreatitis
Small intestine Gallbladder Stores bile until
Enteritis needed
Duodenal ulcer Large intestine
Dysentery
Appendix Colitis Small Site of most digestion
Appendicitis intestine and absorption
Rectum
Anus of nutrients

Food
molecules Large Absorbs some water
intestine and minerals;
prepares waste
Villus
Epithelial cells
Upper digestive tract
Microvilli Lower digestive tract
Smooth
muscle
Capillaries
Nerve fibers
Lymphatic vessel

FIGURE 24.1 The Digestive System Some of the disease conditions that can affect the system are shown in color. For example, mumps
is a viral disease affecting a parotid gland.
? What is the role of villi and microvilli in the small intestine?
 Part IV Infectious Diseases 573

provide a ready source of carbon and energy for microbial growth. When the tooth enamel deteriorates or is otherwise damaged,
Every mouthful of food you ingest feeds not only yourself but the microorganisms can enter the substance of the tooth. Tooth decay,
microbial population in your digestive tract as well. or dental caries, can usually be repaired with dental fillings. If
Most microbes that inhabit the digestive tract live in harmony bacteria reach the root canal, a region of the tooth filled with pulp
with the host, but the balance in this complex ecosystem is deli- (tissue in the tooth that contains blood vessels and nerves), the
cate. A mucous membrane only a single cell layer thick separates tooth must either be removed or the pulp replaced with an inert
the microbial population from some underlying tissues, so damage substance. This replacement procedure—like the region that is
to this layer allows resident microbes to penetrate. Ingested patho- filled—is called a root canal.
gens often have mechanisms to breach intact barriers. Another frequent site of microbial accumulation and infection
is the gingival crevice, the space between the gum and a tooth. In
response to accumulated plaque, the gums become inflamed, a
The Upper Digestive System
condition called gingivitis. The irritated gums may recede from
The upper digestive system includes the mouth, salivary glands, the tooth root, allowing bacteria to reach that region as well, which
esophagus, and stomach. can ultimately result in tooth loss.
Saliva is produced by various salivary glands that empty into
The Mouth and Salivary Glands the mouth. Normally, 1,500 ml of saliva are secreted each day,
The mouth is mainly a grinding apparatus that begins the physical equivalent to approximately four cans of soda pop. The largest of
and chemical digestion of food. The act of chewing allows the these glands, the parotid glands, are notable because the mumps
teeth to grind the food into smaller pieces, while the saliva moist- virus infects them. In addition to saliva’s function in digestion,
ens the food and provides amylase, an enzyme that helps start the it plays an important role in protecting the oral cavity against
breakdown of starches. microbes. Saliva is rich in secretory IgA (the antibodies that pro-
The outer portion of teeth is made up of a hard protective vide mucosal immunity), and it contains antibacterial compounds
substance called enamel (figure  24.2). Proteinaceous material such as lysozyme and lactoferrin. People with poor saliva produc-
from the saliva adheres to the enamel, creating a thin film or tion are subject to severe tooth decay. lactoferrin, p. 337 secretory
pellicle. Various types of bacteria can attach to receptors on the IgA, p. 362 lysozyme, pp. 62, 337
pellicle, colonizing the tooth surface to create a biofilm called Of all the microorganisms entering the mouth, relatively few
dental plaque. Over time, mineral salts deposit in plaque, creating can colonize the surfaces there. Those that do are able to bind
a crusty substance called dental calculus or tartar. Routine brush- specifically to molecules on host tissues or attached microbes,
ing and flossing can remove plaque, but tartar removal requires allowing them to resist the scrubbing action of food and the
professional cleaning. tongue and the flushing effect of salivary flow. They are also able
to withstand the antibacterial effects of saliva. The fact that the
superficial layers of cells of the mucous membranes are constantly
Enamel shed also limits the numbers of microbes in the mouth.
Although over 600 types of bacteria can be found in the
Dentin mouth, members of the genus Streptococcus are among the most
Crown common. The different streptococcal species inhabit different
Pulp
microenvironments. Some preferentially colonize the upper part
Gingival
crevice
of the tongue, others the teeth, and still others the mucosa of
Gingiva (gum)
the cheek. This difference in distribution is due to the fact that
the epithelial cells lining various parts of the mouth have different
receptors. receptors, p. 386
Bone In addition to Streptococcus species—which are obligate
fermenters—obligate anaerobes are also surprisingly common in
Periodontal the mouth. The anaerobes thrive in complex communities, pro-
membrane
Root tected because aerobic species consume available O2. The foul-
Root smelling end products of anaerobic metabolism are associated
canal with halitosis, also known as bad breath.

MicroByte
There can be up to 100 billion bacteria per gram of plaque.
Nerve
Vein
Artery The Esophagus
The esophagus is a collapsible tube about 10 inches long, con-
necting the mouth to the stomach. Peristalsis, the rhythmic con-
FIGURE 24.2 Structure of a Tooth and Its Surrounding tractions of the digestive tract, pushes food to the stomach. The
Tissues esophagus has a relatively sparse microbial population, consisting
? What is the difference between dental plaque and tartar? mostly of bacteria from the mouth and upper respiratory tract.
574 Chapter 24 Digestive System Infections

One reason for the relative lack of microbes is the secretory iron are absorbed as well. In addition, the small intestine absorbs
IgA–containing mucus and saliva that bathe the lining of the fluids. Taking into account the volume of digestive juices, as well
tube. Microbes trapped in those secretions are moved down the as saliva and the fluids ingested as food and drink, this amounts to
esophagus, along with food and liquids, propelled by peristalsis. approximately 9 liters per day! Understandably, disruption in this
The esophagus rarely becomes infected except in individuals with fluid balance can result in diarrhea. active transport, p. 56
AIDS or other immunodeficiencies. AIDS, pp. 633, 695 Few microbes reside in the upper small intestine, because
the flushing action of rapidly passing digestive juices limits their
The Stomach ability to colonize the surface. As the involuntary muscle action
The stomach is an elastic, saclike structure with a muscular wall. of peristalsis propels the intestinal contents toward the large intes-
Its primary function is to break down and store food particles as tine, the movement slows and the bacterial population increases.
they await controlled entry into the small intestine. The gastric The immune system monitors this population with numerous den-
juices are highly acidic, which denatures the proteins in food par- dritic cells sampling the environment and M cells delivering small
ticles. The acidic environment also activates pepsinogen produced amounts of the lumen’s contents to Peyer’s patches. dendritic
in the stomach to form pepsin, a protein-splitting enzyme. The cells cells, p. 340 M cells, p. 358 Peyer’s patches, p. 358
that line the stomach protect themselves from the acid and enzymes
by secreting a thick alkaline mucus. Most bacterial cells cannot
The Large Intestine
survive the hostile environment, however, so a normal empty stom-
ach has few microorganisms. The main function of the large intestine is to absorb water as well
as vitamins produced by the resident microbiota. Because the
small intestine absorbs so much water, only 300 to 1,000 ml of
The Lower Digestive System fluid normally reach the large intestine per day, and most of this is
The lower digestive system includes the small and large intestines, absorbed. The semisolid feces, composed of indigestible material
as well as the pancreas and liver. and bacteria, remain. Infection of the large intestine can interfere
with absorption and stimulate the painful contractions known as
The Small Intestine “stomach cramps.”
As the stomach contents enter the small intestine, digestive fluids Microbes flourish in the large intestine, supported by the
from the pancreas and liver are mixed in. These alkaline fluids abundance of nutrients in undigested and indigestible food
neutralize the stomach acid and also contain bile, an emulsifying material. In fact, bacteria make up about one-third of the fecal
agent that helps break up fats. Substances in bile called bile salts weight, reaching concentrations of approximately 1011 cells per
help the intestine absorb oils, fats, and fat-soluble vitamins. Many gram! Anaerobic bacteria, particularly members of the genus
types of bacteria are killed by bile, but those adapted to live in Bacteroides, make up about 99% of the microbial population.
the intestinal tract resist its bactericidal effects. This has practical The remaining microbes are facultative anaerobes, primarily
applications in the laboratory, because bile salts are used in selec- Escherichia coli and other members of the Enterobacteriaceae.
tive media designed to isolate intestinal bacteria. bactericidal, members of the genus Bacteroides, p. 275 Enterobacteriaceae, p. 265
p. 108 selective media, p. 95 As a community, the intestinal microbiota can degrade a wide
Additional enzymes are produced by intestinal cells. Some are variety of foods. Some high-fiber foods, like beans and broccoli,
secreted into the intestinal lumen (the region inside the tube), but contain substances indigestible by the gastric and intestinal juices
many are permanently attached to the cell membranes. Microbial but readily degraded by intestinal organisms. Microbial breakdown
infections that damage these cells interfere with digestion. of these materials often produces large amounts of gas, causing
If the intestinal tract were a simple pipe, it would have abdominal discomfort and intestinal gas (flatus). Bacterial enzymes
a surface area of less than a bath towel. Although only about can also convert various substances in food to carcinogens and
6 meters in length, the small intestine has an enormous surface therefore may be involved in the development of intestinal cancers.
area, approximately 250 square meters, or the size of a tennis The intestinal microbes are important to human health
court. The inside surface of the intestine is covered with many because they synthesize a number of useful vitamins, including
small, fingerlike projections called villi. Each of these is lined niacin, thiamine, riboflavin, pyridoxine, vitamin B12, folic acid,
with cells that have cytoplasmic projections called microvilli (see pantothenic acid, biotin, and vitamin K. They are also essential
figure  24.1). Bundles of actin filaments make up the structural for normal development of mucosal immunity. At the same time,
core of these projections. This microvilli-coated surface is some- many members of the intestinal microbiota are opportunistic
times referred to as the brush border. At the base of the villi are pathogens that can cause disease if they gain access to other body
pits called crypts. These are small glands that continuously secrete sites such as the urinary tract.
large amounts of enzyme-containing fluids and mucus into the The normal microbiota helps prevent pathogens from coloniz-
lumen. Certain cells in the crypts give rise to new intestinal epithe- ing the large intestine. Antibiotic treatment, especially with broad-
lial cells. Intestinal epithelial cells are completely replaced every spectrum drugs, disrupts the normal microbiota and often results
9 days—one of the fastest turnover rates in the body. actin, p. 73 in mild to severe antibiotic-associated diarrhea. In some cases,
A major role of the small intestine is nutrient and fluid absorp- this is caused by toxin-producing strains of Clostridium difficile,
tion. Cells that line the villi take in amino acids and monosaccha- which readily colonizes the intestine of people whose normal
rides using active transport mechanisms, bringing in sodium ions intestinal microbiota has been reduced by antimicrobial chemo-
(Na+) simultaneously. Fatty acids, vitamins, and minerals such as therapy. Suppression of intestinal microbiota with antibacterial
 Part IV Infectious Diseases 575

medications can also increase susceptibility to other pathogens by the liver. These same processes can also chemically alter and
such as Salmonella enterica. Clostridium difficile–associated disease, remove many medications. Therefore, if the liver is damaged, as
p. 594 Salmonella enterica, p. 592 it might be by a viral infection, lower medication doses might be
needed to prevent an accidental overdose.
The Pancreas
The pancreas is a large organ located behind the stomach. Some
MicroAssessment 24.1
pancreatic cells produce hormones, and others make digestive
enzymes. About 2 liters of pancreatic digestive juices discharge The digestive tract is a complex ecosystem and a major route for
directly into the upper portion of the small intestine each day. pathogens to enter the body. The distribution of bacterial microbiota
in the mouth is governed by different receptors on the epithelium.
These fluids, as well as digestive juices of the small intestine
Bacteria colonizing the tooth surface can create a biofilm called
itself, are alkaline and neutralize the stomach acid as it passes into plaque. People with poor saliva production risk severe tooth decay.
the small intestine. Infections of the esophagus are so unusual that their occurrence
suggests immunodeficiency. The stomach is responsible for acid
The Liver digestion of food and destruction of most microbes before they reach
The liver is a large, dark-red organ located in the upper right por- the intestines. The small intestine carries out the majority of the
tion of the abdomen. One of its roles is to produce bile, which digestion and absorption of nutrients. Undigested material becomes
feces in the large intestine. Metabolic activity of intestinal microbiota
is then concentrated and stored in a saclike structure called the produces vitamins, but it causes gas and can contribute to cancer.
gallbladder. The bile then flows through a system of tubes into Disruption of the normal microbiota through antibiotic treatment can
the upper small intestine. Severe liver disease or obstruction of the result in antibiotic-associated diarrhea.
bile ducts can cause jaundice, a yellow color of the skin and eyes 1. What causes halitosis (bad breath)?
caused by buildup of a bile component (bilirubin) in the blood.
2. What makes the surface area of the small intestine so large,
The liver also inactivates toxic substances that enter the when it is only about 6 meters long?
bloodstream, generally from the digestive tract. For example,
3. Two patients recently had spinal surgery, but one is jaundiced.
ammonia produced by intestinal bacteria and then absorbed into Which one would likely need a lower dose of acetaminophen? +
the bloodstream could reach poisonous levels if not detoxified

UPPER DIGESTIVE SYSTEM INFECTIONS

24.2 ■ Bacterial Diseases of the Signs and Symptoms
Dental caries is usually far advanced before any symptoms
Upper Digestive System develop. Sometimes there is noticeable discoloration, roughness,
Learning Outcomes
or defect, and a tooth can break during chewing. However, the
severe, throbbing pain of a toothache is often the first symptom.
4. Compare and contrast dental caries, periodontal disease, and
acute necrotizing ulcerative gingivitis.
Causative Agent
5. Describe Helicobacter pylori gastritis and how it relates to gastric
ulcers and stomach cancer. Streptococcus mutans and related species are cariogenic (mean-
ing “caries generating”). These Gram-positive cocci live only on
Bacterial diseases of the upper digestive system often involve the teeth and therefore do not colonize the mouth in the absence of
teeth and gums (figure 24.3). Some of these local infections— teeth. Unlike many other bacteria, they thrive in acidic environ-
which often go unnoticed for years—may have consequences for ments (below pH 5), which allows them to survive the conditions
the rest of the body as well. For example, some studies suggest that result from the lactic acid they produce during fermenta-
that chronic gum infections contribute to hardening of the arter- tion. They convert sucrose (table sugar) into extracellular
ies, arthritis, and premature birth. In addition, members of the insoluble polysaccharides called glucans. Glucans are essential
normal oral microbiota can enter the bloodstream during dental for the development of dental caries on smooth tooth surfaces.
procedures and cause subacute bacterial endocarditis. Another polysaccharides, p. 31 fermentation, pp. 134, 147

site of upper digestive infections is the stomach. subacute bacterial

endocarditis, p. 673 Pathogenesis
Formation of dental caries is a complex process. First, oral strepto-
cocci adhere to the pellicle on the tooth to create dental plaque, but
Dental Caries (Tooth Decay) this alone does not lead to caries (figure 24.3). If dietary sucrose
Dental caries (tooth decay) is the most common infectious disease is present, extracellular enzymes produced by S. mutans split the
and the main reason for tooth loss. In the United States, about 60% sucrose into its two monosaccharide components—glucose and
of all teenagers have tooth decay, making it four times more com- fructose. The glucose is polymerized to make glucans, and the
mon than asthma in this group. fructose is fermented, producing lactic acid. The glucans help
576 Chapter 24 Digestive System Infections

Tooth infection Gum infection

Cavity

Bone loss

Filling

Dental plaque

Inflamed
gums

(b)

FIGURE 24.3 Infections of the Teeth and Gums Note the

different types of bacteria in the community forming dental plaque
(shown as a purple color). If plaque hardens between teeth and
gingiva, it can increase the likelihood of periodontal disease, which
5 μm may result in bone loss and loosening or loss of teeth.
? How does brushing away plaque decrease the chance of
(a)
developing cavities?

create an even thicker biofilm by binding a mixed population of rises slowly to neutrality. The delay is due to the storage granules
microbes together and to the tooth, making the plaque impen- made by cariogenic bacteria—polysaccharides in these granules
etrable to saliva. dental plaque, p. 573 can be fermented, generating acid even after the mouth is empty.
Plaque that contains cariogenic microbes can act as a tiny storage granules, p. 66
acid-soaked sponge closely applied to the tooth. In fact, when Both S. mutans and a sucrose-rich diet are required to produce
sugar enters the mouth, the pH of the plaque drops from its normal dental caries on smooth surfaces of the teeth. In deep fissures or
value of about 7 to below 5 within minutes (figure 24.4). This 100- pits, plaque can accumulate in the absence of S. mutans, and tooth
fold increase in acidity begins dissolving the calcium phosphate decay can occur if lactic acid–producing bacteria and fermentable
of the teeth. After food leaves the mouth, the pH of the plaque substances are present.
 Part IV Infectious Diseases 577

7 bristles cannot remove plaque from the deep narrow pits and fis-
Glucose
rinse
sures normally present in children’s teeth, which is where most
childhood tooth decay starts. Caries can be prevented by using a
sealant—a kind of epoxy glue that seals the fissures, kills the bac-
teria in plaque, and prevents bacterial recolonization. Older people
6
have less fissure-related caries, but are prone to receding gums
that expose root surfaces, which become sites for dental caries.
pH

MicroByte
Eliminating sucrose-containing sweets reduces dental caries by 90%.
5

High acidity
in dental plaque Periodontal Disease
Periodontal disease often results from plaque accumulation near
4 the gum margin (periodontal means “around the tooth”). Bacterial
0 10 20 30 40 products in plaque trigger an inflammatory response in the gums
Minutes
and the local tissues, leading to at least some degree of tissue dam-
FIGURE 24.4 Acidity of Dental Plaque Plaque becomes more age. Periodontal diseases include gingivitis (swelling and redness
acidic after rinsing the mouth with a glucose solution. Tooth enamel of the gums) and chronic periodontitis, a destructive response
begins to dissolve at about pH 5.5.
that progressively damages the structures that support the teeth
? Would you expect this dip in pH if Streptococcus mutans (see figure 24.3). Chronic periodontitis is an important cause of
metabolized sugar via aerobic respiration rather than
tooth loss from middle age onward.
fermentation?

Epidemiology Signs and Symptoms

Dental caries is worldwide in distribution, but the incidence var- Gingivitis is marked by gums that are tender and bleed easily.
ies markedly depending mainly on intake of dietary sucrose and This can occur within days of plaque accumulation and typically
access to preventive dental care. Genetics also plays an important resolves once plaque is removed.
role because some people inherit resistance to the disease. The Chronic periodontitis is a long-term response to plaque and is
incidence of dental caries peaks during teen years and falls off characterized by bad breath, red shiny gums that bleed easily, and
with age. This is probably because the pits and fissures on the loosening of the teeth. The base of the teeth becomes discolored—
tooth—ideal sites for bacterial growth—wear down with time. ranging from yellowish to black—and the gums recede. Exposed
roots of the teeth are susceptible to dental caries.
Treatment and Prevention
Treatment of dental caries requires drilling out the cavity, filling Causative Agent
the defect with material such as amalgam (an alloy of mercury and Periodontal disease is associated with dental plaque that forms at
some other metal), and restoring the contour of the tooth. the point where the gum joins the tooth. The plaques are typically
The most important method for controlling dental caries is quite different from those associated with dental caries in that
restricting dietary sucrose and other refined carbohydrates. This most members of the microbial community are Gram-negative
reduces Streptococcus mutans colonization as well as acid produc- anaerobes. An association of three types bacteria—Porphyromonas
tion by cariogenic plaque. However, the quantity of sugar in the gingivalis, Treponema denticola, and Tannerella (Bacteroides)
diet is not the most important factor leading to dental caries—the forsythia—seem to play an important role. However, hundreds of
frequency of eating sugary foods and the length of time the food kinds of bacteria have been identified in plaques associated with
stays on the teeth are more critical. In fact, chewing paraffin or periodontal disease, many of which have not been cultivated, so
sorbitol-sweetened gum reduces dental caries, probably because it other microbes are likely important as well.
increases the flow of saliva.
Trace amounts of fluoride prevent dental caries by making Pathogenesis
tooth enamel harder and more resistant to dissolving in acid. In As plaque (and tartar) accumulates at the gum margin—especially
the United States, more than half the population is supplied with in hard-to-clean areas between the teeth—it gradually extends into
fluoridated public drinking water, resulting in a 60% reduction the gingival crevice. Bacterial products incite an inflammatory
in dental caries. This saves over $4.5 billion in dental costs each response, leading to the characteristic symptoms of gingivitis.
year. In areas where fluoridated drinking water is not available, If the level of plaque remains small, the host response limits the
fluoride tablets or solutions can be used. To have optimum effect, process. tartar, p. 573
children should begin receiving fluoride before their permanent In some cases, the inflammatory response does not control the
teeth erupt. Fluoride applied to tooth surfaces in the form of microbial population, leading to the tissue destruction associated
mouthwashes, gels, or toothpaste is generally less effective. with chronic periodontitis. The process appears to involve many
Toothbrushing and flossing remove plaque, reducing the factors, including the microbial population and host factors. Tissue-
incidence of dental caries by about half. Unfortunately, toothbrush degrading enzymes released by the plaque microbes weaken the
578 Chapter 24 Digestive System Infections

gingival tissue and cause the gingival crevice to widen and deepen.
This allows the plaque to spread further toward the root of the tooth.
The toll-like receptors of surrounding tissues detect the lipopolysac-
charide of the Gram-negative outer membrane (endotoxin), causing
the release of pro-inflammatory cytokines. Some of these evoke
such a strong inflammatory response that nearby tissues are damaged.
With progressive tissue damage, the membrane that attaches the
tooth root to the bone weakens, and the bone gradually softens.
The tooth becomes loose and may fall out. toll-like receptors, p. 342

Epidemiology (a)
Periodontal disease such as gingivitis can begin in childhood.
After age 65, almost 90% of individuals have some degree of
chronic periodontitis. Smokers and those with underlying defects
in acquired or innate immunity (including those with AIDS) often
have severe periodontitis that leads to tooth loss.

Treatment and Prevention

Periodontal disease can be treated in its early stages by cleaning
out the inflamed gingival crevice and removing plaque and tartar.
In advanced cases, minor surgery is usually required to expose
and clean the roots of the teeth. Careful flossing and toothbrushing
can prevent periodontal disease, especially when combined with (b) 3 μm
twice-yearly polishing and removal of tartar at a dental office. FIGURE 24.5 Acute Necrotizing Ulcerative Gingivitis (ANUG)
(a) Red, swollen gums with loss of tissue, especially between the
teeth. (b) Gram stain of exudate showing a spirochete and rod-
Acute Necrotizing Ulcerative Gingivitis shaped bacteria.
Acute necrotizing ulcerative gingivitis (ANUG) is a severe, acute ? What aspects of methamphetamine abuse increase the risk of
condition distinct from other forms of periodontitis (figure 24.5). developing ANUG?
The disease was first called trench mouth because it was common
among soldiers living in trenches during World War I, unable to
care for their teeth and gums. Since 2005, dentists have noticed Methamphetamine use often lowers saliva production, lead-
a dramatic increase in ANUG cases associated with methamphet- ing to a dry mouth and increased risk of biofilm formation, tooth
amine use, coining the term “meth mouth” as a form of ANUG. decay, and ANUG. Tooth grinding, also associated with metham-
Meth mouth is further complicated by rampant dental caries. phetamine use, increases the risk of fracturing teeth weakened by
severe decay and gingivitis.
Signs and Symptoms
ANUG is characterized by bleeding painful gums, abscessed and Epidemiology
broken teeth, and extremely foul breath. ANUG can occur at any age in association with poor oral hygiene,
particularly when combined with poor nutrition, high sugar diet,
Causative Agent chronic stress, and immunodeficiency. Methamphetamine use
The suspected causative agent of ANUG is an oral, not yet culti- appears to intensify the problems.
vated spirochete of the Treponema genus. The organism probably
acts synergistically with other anaerobic species, with which it Treatment and Prevention
is always found. The disease is associated with heavy growth of Most bacteria at the gingival crevice are anaerobes, so hydrogen per-
anaerobes at the gum line. Only about 50% of oral microbiota oxide treatment rapidly relieves ANUG symptoms. Long-term cures
members have been grown in culture, so their role in ANUG is include removing plaque and tartar. Prevention begins with daily
poorly understood. the genus Treponema, p. 276 brushing and flossing, and twice-yearly professional cleaning. The
main features of teeth and gum infections are presented in table 24.1.
Pathogenesis
The spirochetes and the other anaerobes are presumed to act
together to destroy tissues in the oral cavity, but the precise Helicobacter pylori Gastritis
mechanisms are unknown. Plaque is always present, but its bacte- Many people have gastritis, meaning inflammation of the stomach,
rial composition shows much larger numbers of spirochetes and without even knowing it. The association of the causative agent to
other anaerobes than are present in chronic periodontal disease. ulcers was demonstrated in the 1980s when Barry Marshall, one
The spirochetes invade the tissue, causing necrosis and ulceration, of the scientists who discovered the organism, intentionally drank
mainly of the gums between the teeth. a culture of Helicobacter pylori.
 Part IV Infectious Diseases 579

TABLE 24.1 Important Infections of the Teeth and Gums

Acute Necrotizing
Dental Caries Periodontal Diseases Ulcerative Gingivitis

Signs and None until advanced disease. Gingivitis characterized by tender Bleeding painful gums, abscessed
symptoms Late: discoloration, roughness, bleeding gums; chronic periodontitis and broken teeth, and extremely
broken tooth, throbbing pain characterized by bad breath, red shiny foul breath
gums that bleed easily, loose teeth, and
exposed discolored tooth roots
Incubation period 1 to 24 months before cavity is Gingivitis—days; periodontitis—months Undetermined
detectable or years
Causative agent Dental plaque, particularly when Microbes in dental plaque; an association Probably a spirochete of the genus
populated with Streptococcus of Porphyromonas gingivalis, Treponema Treponema acting with other
mutans denticola, and Tannerella forsythia seem anaerobes
to play an important role in periodontitis.
Pathogenesis Bacteria in plaque produce acid Plaque formed at the gum margins causes The spirochetes and certain other
from dietary sugars; slowly dissolves the inflammatory response associated anaerobes act synergistically to
the calcium phosphate composing with gingivitis. If the plaque extends destroy tissues. The spirochetes
the tooth; sucrose is critical for into the gingival crevices, the strong invade tissue, causing necrosis and
cariogenic plaque formation. inflammatory response damages tissues ulceration.
that support the teeth, causing chronic
periodontitis.
Epidemiology Worldwide distribution, incidence Gingivitis can begin in childhood; All ages are susceptible in
depending on dietary sucrose, periodontitis primarily affects older association with poor oral hygiene,
natural or supplemental fluoride. people, particularly smokers and malnutrition, immunodeficiency, or
The young are more susceptible immunodeficient individuals. methamphetamine use.
than the old.
Treatment Restriction of dietary sucrose, Avoid buildup of plaque. Surgical Avoid buildup of plaque. Antibiotic
and prevention supplemental fluoride, mechanical treatment in severe cases of periodontitis treatment, followed by removal of
removal of plaque, sealing pits and to clean tooth roots. plaque and tartar.
fissures in children’s teeth.

Signs and Symptoms required for H. pylori to colonize the stomach. Non-motile strains
Most Helicobacter pylori infections are asymptomatic. Early are removed with mucus turnover, and urease-deficient ones can
in infection, however, the signs and symptoms may range from cause disease only if injected directly into the mucus layer.
belching to vomiting. When the infection results in peptic ulcers Virulent H. pylori strains produce CagA, a protein they inject
of the stomach and duodenum (peptic meaning “caused by diges- into host cells that changes the shape and surface characteristics of
tive juices”), the patient may experience localized abdominal pain, the cells. Evidence suggests that these changes represent a prelude
tenderness, and bleeding. Stomach cancer can also develop. to cancer. Another bacterial product, VacA, acts on mucosal cells
to promote flow of urea into the stomach. This protein also might
damage the mucus-producing epithelial cells.
Causative Agent
Bacterial products from H. pylori cells growing near the
Helicobacter pylori is a short, curved, Gram-negative, microaero- epithelium incite an inflammatory response in the wall of the
philic bacterium. It has multiple polar flagella that are unusual stomach. As the epithelial cells are damaged by a combination
because they are covered by sheaths. the genus Helicobacter, p. 276
of products released from the responding neutrophils and toxins
produced by H. pylori, mucus production decreases. The thinning
Pathogenesis of the protective mucus layer and damage to local cells probably
Helicobacter pylori cells survive the acidic environment of the accounts for the development of peptic ulcers.
stomach by (1) producing urease, an enzyme that converts urea H. pylori infections persist for years, often for life. The
to ammonia, thereby creating an alkaline microenvironment, and outcome of the infection is quite variable. Only about one in six
(2) burrowing within the mucus layer that coats the stomach lin- infected persons develops ulcers. A small percentage of chroni-
ing (figure 24.6). Urea is normally found in gastric juices because cally infected people develops stomach cancer, but more than 90%
it is released as proteins are degraded. H. pylori cells use their of those with stomach cancer are infected.
flagella to move through the mucus, following the pH gradient
from the acidic gastric lumen to the nearly neutral underlying epi- Epidemiology
thelial cells. The bacterial cells then attach to the mucus-secreting Overall, about one in five adults in the United States is infected
epithelium or multiply adjacent to it. The ability to both move with H. pylori, but the incidence increases with age, reaching almost
away from the acidic lumen and produce ammonia from urea are 80% for those over age 75. Infections tend to cluster in families.
580 Chapter 24 Digestive System Infections

FIGURE 24.6 Helicobacter pylori Pathogenesis Urease

Neutrophils are early responders to invasion, CO(NH2)2 + H2O CO2 + 2 NH3
followed by other cells such as lymphocytes. Urea Ammonia
? How does urease allow
H. pylori to live in the stomach?

1 Helicobacter pylori 2 Bacteria 3 Thinning of mucus layer

penetrate and
Acidic environment due to mucosal damage
multiply in
of stomach mucus layer Epithelium destroyed
by gastric acid

Mucus layer Mucus- Ulcer

secreting cell
Epithelium
lining the
stomach

Basement
membrane
Connective
tissue Neutrophil Plasma Lymphocyte Red blood cell
cell Dilated capillary
Capillary
Red blood cell Inflammatory response

Helicobacter survives the acidic environment Bacteria attach. Bacterial Inflammation and toxins damage epithelial cells,
of the stomach by secreting urease, which proteins change the surface decreasing mucus production; acidic stomach juices
breaks urea into carbon dioxide and ammonia. of epithelial cells and damage damage the exposed tissue, causing a peptic ulcer.
Ammonia neutralizes stomach acid. mucus-secreting cells.

Transmission of H. pylori probably occurs by the fecal-oral

TABLE 24.2 Helicobacter pylori Gastritis
route, and the bacteria have been found in well water. Flies that
Signs and Infections are often asymptomatic, but have landed on feces are also capable of transmitting the organ-
symptoms peptic ulcers can cause abdominal pain, ism. Infection rates are highest in low-socioeconomic groups.
tenderness, and bleeding. Stomach cancer
can develop.
Treatment and Prevention
Incubation period Usually undetermined
Helicobacter pylori infections can be treated with a combination
Causative agent Helicobacter pylori, a curved, Gram- of two antibiotics and a medication that inhibits stomach acid pro-
negative, microaerophilic bacterium, with
multiple polar flagella covered by sheaths
duction. This usually results in clearing of the gastritis and healing
of any ulcers. There are no proven preventive measures. The main
Pathogenesis H. pylori cells survive stomach acidity by
features of H. pylori gastritis are shown in table 24.2.
producing urease and burrowing within
the stomach's mucus coating. The bacterial
cells attach to and transfer CagA to MicroAssessment 24.2
epithelial cells, causing changes that may
The pathogenesis of tooth decay depends on both dietary sucrose
be a prelude to cancer. The VacA they
produce increases flow of urea into the
and acid-forming bacteria in biofilms on teeth. Periodontal
stomach. The inflammatory response to disease is an inflammatory disease associated with plaque at the
infection and bacterial toxins may account gum margin and can lead to loosening of teeth. ANUG destroys
for peptic ulcers. Cancer rarely develops, gingival tissue. Chronic infection by Helicobacter pylori is a key
but most people with stomach cancer are factor in the development of stomach and duodenal ulcers and
infected with H. pylori. stomach cancer.
Epidemiology Probably fecal-oral transmission. Incidence 4. Why are new cavities less common, but loss of teeth more
increases with age. common, in people over age 65?
Treatment No proven preventive. Treated with a 5. What enzyme produced by Helicobacter pylori helps it survive
and prevention combination of two antibiotics and a in stomach acid?
medication that inhibits stomach acid 6. Do you agree or disagree with the statement “Helicobacter
production. pylori causes stomach cancer.” Explain. +
 Part IV Infectious Diseases 581

CASE PRESENTATION
The patient was a 35-year-old man who con- 4. It took a long time for doctors to accept also provokes a strong immune response.
sulted his physician because of upper abdomi- that this condition had an infectious The protein VacA promotes the flow of
nal pain. The pain was described as a steady etiology. Why? urea into the stomach. Once established,
burning or gnawing sensation, like a severe H. pylori infections persist for years and
hunger pain. Usually it came on 1* to 3 hours Discussion often for a lifetime. It is not known why
after eating, sometimes waking him from 1. This patient had a duodenal ulcer. The some people develop gastric or duode-
sleep. Generally, it was relieved in a few min- ulcer had penetrated deeply beyond the nal ulcers and others do not. Both host
utes by food or antacid medicines. mucosa, involving small blood vessels and and bacterial factors are almost certainly
On examination, the patient appeared causing bleeding. This was apparent from involved. For example, strains of H. pylori
well, without evidence of weight loss. The the clot seen at endoscopy and the positive isolated from peptic ulcer patients tend to
only positive finding was tenderness slightly test for blood in the stool. be more virulent than those from patients
to the right of the midline in the upper part of who just have gastritis; patients with blood
2. Microscopic examination of the biopsy
the abdomen. A test of the patient’s feces was group O have more receptors for the bac-
showed curved bacteria, confirmed by
positive for blood. The remaining laboratory terium and a higher incidence of peptic
culture to be Helicobacter pylori.
tests were normal. ulcers than do other people. Stomach acid
Endoscopy, a procedure that uses a long, 3. Helicobacter pylori cells enter the gas- and peptic enzymes probably play a role
flexible fiber-optic device passed through the trointestinal tract by the fecal-oral route. in ulcer formation by acting on damaged
mouth, showed a patchy redness of parts of the In the stomach, they escape the lethal epithelium unprotected by normal mucus.
stomach lining. A biopsy was taken. The endos- effect of gastric acid because they pro-
4. Claude Bernard, a scientist of Pasteur’s
copy tube was passed through the pylorus and duce urease. Highly motile, they enter the
time, put it this way: “It is that which we
into the duodenum. About 2 cm into the duode- gastric mucus and follow a gradient of
do know which is the greatest hindrance to
num, there was a lesion 8 mm in diameter that acidity ranging from pH 2 in the gastric
our learning that which we do not know.”
lacked a mucous membrane and appeared to be juices to pH 7.4 at the epithelial surface.
In 1983, when Dr. Barry J. Marshall pro-
“punched out.” The base of the lesion was red Mutant strains that lack the ability to
claimed before an international gathering
and showed an adherent blood clot. After the produce urease are infectious only if they
of infectious disease experts that a bacte-
endoscopy, a portion of material obtained by are introduced directly into the mucus
rium caused stomach and duodenal ulcers,
biopsy was placed on urea-containing medium. layer. Multiplication occurs just above
everyone “knew” it could not be true
Within a few minutes, the medium began to the epithelial surface, but some of the
because no organism was thought to exist
turn color, indicating a developing alkaline pH. bacteria attach to the epithelial cells and
that could survive stomach acidity and
cause a loss of microvilli and thickening
1. What is the patient’s diagnosis? enzymes. Indeed, almost everyone already
at the site of attachment. An inflammatory
2. What would you expect microscopic “knew” the cause of ulcers to be psycho-
reaction develops beneath the affected
examination and culture of the gastric somatic. There is much still to be learned
mucosa. Two genes, cagA and vacA, cor-
mucosa biopsy to show? about the cause of ulcers, however, and
relate with virulence. The bacteria inject
Bernard’s statement remains relevant.
3. Outline the pathogenesis of this patient’s the protein CagA into host cells, causing
disease. the cells to elongate and spread out. CagA

24.3 ■ Viral Diseases of the Upper Herpes Simplex (Cold Sores)

Digestive System Herpes simplex (cold sores) is an extremely widespread disease
with many symptoms. In its most common form, it begins in the
Learning Outcome mouth and throat where it causes cold sores (also called fever
6. Compare and contrast herpes simplex (cold sores) and mumps. blisters). Involvement of the esophagus suggests AIDS or other
immunodeficiency. Although the disease is usually insignificant,
In this section, we focus on two viral diseases that have dramatic it can have tragic consequences in newborn infants or people with
signs and symptoms involving the upper digestive system. Herpes immunodeficiency.
simplex (cold sores) is characterized by painful oral ulcers.
Mumps causes enlarged and painful parotid glands.
Some viral diseases involve the upper digestive system but Signs and Symptoms
produce more dramatic symptoms elsewhere in the body. For The initial signs and symptoms include fever and small blisters
example, measles causes an obvious skin rash and respiratory in the mouth. The blisters break within a day or two, producing
symptoms, and it causes Koplik spots in the mouth. Chickenpox superficial ulcers that can be so painful they make it difficult to
causes a skin rash and oral blisters. Infectious mononucleosis eat or drink. Although the lesions heal without treatment within
causes impressively enlarged lymph nodes and spleen and may about 10 days, the virus persists as a latent infection; the affected
give rise to oral ulcers and bleeding gums. chickenpox, p.  534 person may suffer the recurrent cold sores, also known as herpes
infectious mononucleosis, p. 680 measles, p. 537  simplex labialis (the word labialis indicates location of the lesions
582 Chapter 24 Digestive System Infections

of some U.S. populations, usually resulting in mild symptoms.

Approximately one in three Americans suffers from cold sores.
The virus is transmitted primarily by close physical contact, but
can survive for several hours on plastic and cloth, so transmission
by fomites is also possible. The greatest risk of infection is from
contact with lesions or saliva from patients within a few days of
Intranuclear disease onset, because at this time large numbers of virions are
inclusion present. Even the saliva of asymptomatic people can be infectious,
body however, posing a risk to dentists and other healthcare workers.
Infected fomite, p. 440
epithelial cell HSV can infect almost any body tissue. For example, herpetic
Giant cell
whitlow, a painful finger infection, is not uncommon among nurses.
Wrestlers can develop infections at almost any skin site because
Neutrophils
saliva containing HSV can contaminate wrestling mats and get
FIGURE 24.7 Herpes Simplex Labialis (Cold Sores or Fever rubbed into abrasions. Blindness can result if the virus is rubbed into
Blisters) The photomicrograph is of stained material from a herpes the eye. Although uncommon, HSV is the most frequently identified
simplex lesion. It shows a multinucleated giant cell and intranuclear cause of sporadic viral encephalitis, a serious brain disease.
inclusion bodies. The pink areas within the epithelial cell nuclei
are inclusion bodies, the sites of viral replication.
Treatment and Prevention
? How do the signs and symptoms of recurrent herpes simplex differ
from those of the primary infection? Medications such as acyclovir and penciclovir target HSV DNA
polymerase and are useful for treating severe cases and for pre-
venting disabling recurrences. They do not affect the latent virus,
however, and therefore cannot cure the infection. Sunlight expo-
on the lips) (figure 24.7). The signs and symptoms of recurrences sure can trigger disease recurrence, so sunscreens are sometimes
are usually less severe than those of initial infection. They include a helpful preventive. Some of the main features of herpes simplex
tingling, itching, and burning on the lips, followed by blisters and are shown in table 24.3. DNA polymerase, p. 166 acyclovir, p. 475
painful ulcerations, which usually heal within 7 to 10 days.

Causative Agent
Cold sores are caused by herpes simplex viruses (HSV), envel- TABLE 24.3 Herpes Simplex
oped viruses containing double-stranded, linear DNA. There are
two types of the virus—HSV-1 and HSV-2. Most oral infections Signs and Initial infection: fever, severe throat pain,
symptoms ulcerations of the mouth and throat.
are due to HSV-1; HSV-2 usually causes genital infections. Both
Recurrences: itching, tingling, or pain
forms persist throughout life as latent viruses that reactivate peri- usually localized to the lip, followed by
odically. genital herpes, p. 630 latent virus, p. 322 blisters that break leaving a painful sore,
which usually heals in 7 to 10 days.
Pathogenesis Incubation period 2 to 20 days
HSV multiplies in the epithelium of the mouth or throat, and Causative agent Herpes simplex virus (HSV), usually type 1
destroys the cells. Some epithelial cells fuse together, producing
Pathogenesis The virus multiplies in the epithelium and
large, multinucleated giant cells. The nucleus of an infected cell destroys the cells. Blisters contain large
typically contains a deeply staining area called an intranuclear numbers of infectious virions. An immune
inclusion body—the site of viral replication (figure  24.7). The response limits the infection, but non-
characteristic blisters contain many infectious virions. Although infectious HSV DNA persists in sensory
nerves. This DNA becomes the source of
an immune response develops and quickly limits the infection,
infectious virions that are carried to the
some virions enter the sensory nerves in the area. skin or mucous membranes, usually of the
Viral DNA persists in nerve cells in a latent form— lip, causing recurrent sores.
non-infectious and non-replicating. This latent virus can occasion- Epidemiology Widespread virus, transmitted by
ally reactivate. The reactivated virus is carried by the nerves to close physical contact. The saliva of
skin or mucous membranes where it produces recurrent disease. asymptomatic individuals can be infectious.
Stresses that can precipitate recurrences include menstruation, Treatment Acyclovir, penciclovir, and similar
sunburn, and any illness associated with fever. and prevention medications that target HSV DNA
polymerase can shorten the duration of
Epidemiology the symptoms or prevent recurrences.
Sunscreens can prevent recurrences due to
The initial infection with HSV typically occurs during child- ultraviolet exposure.
hood. The virus is extremely widespread and infects up to 90%
 Part IV Infectious Diseases 583

Mumps post-pubertal boys and men are complicated by orchitis—a rapid,

intensely painful swelling of one or both testicles to three to four
Mumps is an acute viral illness that often affects glands such times their normal size. Atrophy (shrinkage) of the involved
as the parotids. Formerly common in the United States, the testicles commonly develops after recovery from the illness, and
disease is now relatively rare because of routine childhood in rare cases causes sterility. In women and post-pubertal girls,
immunization. Outbreaks do occur, however, mostly due to ovarian involvement occurs in about one of 20 cases and is
waning immunity in college students and other young adults. manifested by pelvic pain.
A large outbreak in 2006 resulted in over 6,000 reported cases.
Causative Agent
Signs and Symptoms
Mumps virus is an enveloped single-stranded RNA virus. It is a
The onset of mumps is marked by fever, loss of appetite, and member of the paramyxovirus family, a group that includes the
headache. These symptoms are typically followed by painful rubeola and respiratory syncytial viruses. Only one antigenic type
swelling of one or both parotid glands (figure  24.8). Spasm of the mumps virus is known.
of the underlying muscle makes it difficult to chew or talk,
perhaps giving rise to the name of the disease (mump means Pathogenesis
“to mumble” or “whisper”). Symptoms usually disappear in
The mumps virus enters the body when virus-laden droplets of
about a week.
saliva are inhaled. It reproduces first in the upper respiratory
Painful parotid swelling is characteristic of mumps, but
tract, then spreads throughout the body in the bloodstream.
symptoms can arise elsewhere in the body with or without parotid
Symptoms begin only after tissues such as the parotid glands,
swelling. For example, headache and stiff neck indicate that the
meninges, pancreas, ovaries, or testicles are infected. Because
virus is causing meningitis—infection of the coverings of the
of this, the incubation period is relatively long, generally 15
brain. Pregnant women with mumps often miscarry. Rare but seri-
to 21 days.
ous consequences of mumps, such as death from brain infection,
In the parotid salivary glands, the virus multiplies in the epi-
are most likely to occur in older people. Sudden-onset deafness
thelium of ducts that convey saliva to the mouth. This destroys
due to asymptomatic infection has also been reported. meningitis,
these cells, releasing enormous quantities of virus into the saliva
p. 643
and eliciting a strong inflammatory response, which causes the
Mumps symptoms are generally much more severe in indi-
severe swelling and pain characteristic of the disease. A similar
viduals past the onset of puberty. About one-quarter of cases in
sequence of events occurs in the testicles, where the virus infects
the system of tubules that convey the sperm. The swelling and
pressure often impair the blood supply, which can lead to hem-
orrhage and death of testicular tissue. Kidney tubules are also
infected, and the virus can be cultivated from the urine for 10 or
more days following the onset of illness.

Epidemiology
Humans are the only natural host of mumps virus. There is
only one antigenic type, so infection confers lifelong immunity.
Individuals sometimes claim to have had mumps more than once,
probably because other infectious and non-infectious diseases can
cause parotid swelling.
The mumps virus is often spread by individuals who have
asymptomatic infections but secrete the virus in their saliva and
continue to mingle with other people. In symptomatic patients, the
virus can be present in saliva from almost a week before symp-
toms appear to 2 weeks afterward. Peak infectivity however, is
from 1 to 2 days before parotid swelling until the gland begins to
return to normal size.

Treatment and Prevention

There is no effective treatment for mumps. However, an effec-
tive attenuated vaccine has been available in the United States
since 1967. It is part of the measles, mumps, rubella, and vari-
FIGURE 24.8 A Child with Mumps The swelling directly below cella vaccine (MMRV). Figure 24.9 shows how the incidence
the earlobe is due to enlargement of the parotid gland. of mumps has generally declined, although it increased in the
? When might mumps result in sterility? 1980s because of cuts to funding for vaccinations. Mumps is a
584 Chapter 24 Digestive System Infections

50 7,000 TABLE 24.4 Mumps

45 6,000 Signs and Fever, headache, loss of appetite, followed
symptoms by painful swelling of one or both parotid

Number of cases reported

glands. Meningitis can occur. Painful
Reported cases per 100,000 population

40 5,000
enlargement of the testicles in men, pelvic
pain in women.
35 4,000
Incubation Generally 15 to 21 days
30 3,000 period
Causative agent Mumps virus, a single-stranded RNA virus of
25 2,000 the paramyxovirus family
Pathogenesis The virus initially replicates in the upper
20 1,000
respiratory tract, then spreads throughout
the body in the bloodstream. In the parotid
15 0 salivary glands, the virus multiplies in the
2003 2005 2007 2009 cells that line the ducts. The inflammatory
10 Year response to cell destruction causes the
swelling and pain.
5
Epidemiology Humans are the only source of the virus.
Infections are often asymptomatic, so the
0 disease can be spread unknowingly.
1973 1978 1983 1988 1993 2003 2008
Year Treatment No antiviral therapy is available. An
and prevention effective attenuated vaccine is available.
FIGURE 24.9 Reported Cases of Mumps per 100,000
Population, United States, 1973–2009 Mumps vaccine was
licensed in 1967. Two doses of MMR vaccine were recommended in
1989 following an outbreak of over 20,000 cases in 1986–1987. An
epidemic among college students resulted in more than 6,000 cases MicroAssessment 24.3
of mumps in 2006. Another outbreak of about 2,000 cases occurred in Herpes simplex (cold sores) is characterized by acute infection
2009–2010, prompting one New York county to begin offering a third followed by lifelong latency and the possibility of recurrent disease.
dose of MMR vaccine. Infectious virus is often present in saliva in the absence of symptoms.
? What events or factors might have contributed to the 2006 Mumps virus infections characteristically cause enlargement of the
mumps epidemic among college students? parotid glands, but they can involve the brain, testicles, and ovaries,
and cause miscarriages.
7. In what type of cell does HSV-1 persist?
8. Why is mumps virus a good candidate for worldwide
good candidate for eradication because it infects only humans, eradication?
latent infections do not occur, and there is only one sero-
9. Why would acyclovir not cure HSV infections even though it
type. The main features of mumps are presented in table 24.4. prevents recurrent cold sores? +
eradication, p. 447

LOWER DIGESTIVE SYSTEM INFECTIONS

One out of five children in developing countries dies of diarrhea 24.4 ■ Bacterial Diseases of the
before the age of 5. Most of the 5 million individuals who die of
diarrhea each year are infants, but no age group is spared. Fatal Lower Digestive System
cases are less common in the United States, but millions of diar-
rhea cases still occur each year. Learning Outcomes
Microbes infecting the digestive tract can cause intestinal 7. Describe the general characteristics of bacterial intestinal diseases.
symptoms, and so can foodborne microbial toxins (foodborne 8. Compare and contrast cholera, shigellosis, the various types
intoxication). This chapter focuses on intestinal infections, and of E. coli gastroenteritis, campylobacteriosis, and Clostridium
chapter 31 gives examples of foodborne intoxications. Tapeworm difficile–associated disease.
and roundworm infections are discussed in chapter 12. Information
about other helminth infections can be found at the text website Diarrheal illness is a common result of bacterial infection of the
(www.mhhe.com/nester7). foodborne intoxication, p.  756 intestinal tract. Bacteria can also use the intestines as an entry to
helminth infections, p. 295 the rest of the body, thereby causing other types of illness.
 Part IV Infectious Diseases 585

General Characteristics Toxin production

Although many different bacteria can infect the intestinal tract,
Cl–
the diseases they cause share some general characteristics. Na+
H2O
Cytotoxins cause
Signs and Symptoms cell death. Toxins
Enterotoxins absorbed into the
The all-too-familiar signs and symptoms of intestinal diseases increase secretion bloodstream result
include diarrhea, loss of appetite, nausea and vomiting, and some- of water and in systemic effects.
times fever. The incubation period is typically a day or two, but electrolytes.
varies according to the dose consumed. Some physicians often
refer to diarrheal disease as gastroenteritis (gastro-, “stomach,”
entero-, “intestine,” -itis, “inflammation”), whereas others prefer
Alterations in the host cells
the term “stomach flu.” (Note “stomach flu” has no relationship
to “the flu,” or influenza.) Pedestal
Attachment and effacing
Diarrhea can be copious and watery (“the runs”) when infec- (A/E) lesions formed after
tion involves the small intestine. Large intestine invasion causes bacterium injects various effector
smaller amounts of diarrhea (“the squirts”) that contains mucus, proteins. One protein functions
as a receptor for the bacterium.
pus, and sometimes blood. The name dysentery is given to diar- Another induces rearrangement
rheal illnesses when pus and blood are present in the feces. A few of actin filaments, resulting in the
bacterial pathogens first establish an intestinal infection and then formation of a pedestal under
Inject effector the bacterium.
spread systemically. This causes enteric fever, characterized by proteins
systemic signs such as shock. shock, p. 394

Cell invasion
Causative Agent
Members of the family Enterobacteriaceae are among the most
common causes of bacterial diarrhea. These include species of Bacterium is engulfed and
multiplies within host cell.
Shigella and Salmonella, and some strains of E. coli. Other causes An effector protein injected by
are Vibrio cholerae and Campylobacter jejuni. In individuals with the bacterium induces the
predisposing conditions, Clostridium difficile causes diarrhea. engulfment by causing
rearrangement of host cell actin.

Pathogenesis
The pathogenesis of bacterial intestinal disease involves a number FIGURE 24.10 Common Mechanisms of Pathogenesis of
Intestinal Pathogens Invasion or cell damage elicits a strong
of different mechanisms. Various strains within the same species inflammatory response that also contributes to intestinal
can differ in the way they cause disease, and the same strain can pathology.
use more than one mechanism. Moreover, many of the responsible
? Why is adhesion generally a prerequisite for pathogenicity?
genes are on plasmids, phages, or other mobile genetic elements,
which can be transferred from one species to another by horizontal
gene transfer. mobile genetic elements, p. 208
Attachment to the intestinal surface is often a prerequisite for of microvilli on the intestinal cell surface with a thick structure—
infection, and this typically involves adhesins on pili. Additional a “pedestal”—under the bacterium. The characteristic damage
common mechanisms of pathogenesis include the following caused by these bacteria is called attaching and effacing (A/E)
(figure 24.10): adhesins, p. 386 lesions. type III secretion systems, p. 386 effector proteins, p. 387
■ Toxin production. Toxins involved in intestinal infections ■ Cell invasion. Some pathogens use type III secretion systems
fall into two groups: enterotoxins, which cause water and to deliver effector molecules that induce the intestinal epithe-
electrolytes to flow from intestinal cells; and cytotoxins, lial cells to engulf the bacteria (see figure 16.6). Once inside,
which cause cell death. Some types of cytotoxins produced in they can multiply.
the intestine can be absorbed into the bloodstream, resulting Severe watery diarrhea occurs as a result of rapid loss of water and
in systemic effects. exotoxins, p. 391 electrolytes from intestinal cells. This causes severe dehydration,
■ Alterations in intestinal epithelial cells. Some pathogens as the intestinal cells draw fluid from the bloodstream. Because of
alter the characteristics of intestinal epithelial cells, using the reduced blood volume, the flow may not be sufficient to keep
type III secretion systems to deliver effector proteins to those vital organs working properly, a potentially fatal situation.
cells (see figure  16.5). For example, certain strains of E. coli Invasion or cell damage elicits a strong inflammatory
inject protein molecules that assemble in the host plasma response. This primarily occurs in the large intestine and results
membrane. The bacteria then use these molecules as recep- in the pus and blood in the feces that characterize dysentery.
tors for closer attachment. After that, the bacterial cells inject Proteins injected by type III secretion systems can also elicit an
proteins that rearrange actin filaments, resulting in replacement inflammatory response.
586 Chapter 24 Digestive System Infections

PERSPECTIVE 24.1
Ecology of Cholera
The ecology of cholera is fascinating but still conjugation. Various strains of V. cholerae, ents cause explosive growth of phytoplankton.
incompletely understood. The O139 Vibrio most of which are not pathogenic for humans, These findings could explain (1)  how new
cholerae strain arose in India in the fall of live in the coastal seas around the world, pandemic strains could arise through genetic
1992 from a serotype O1 strain, the same largely in association with zooplankton. These interchange, (2) an association between chol-
serotype as the pandemic and American Gulf zooplankton can sometimes also harbor the era and climatic changes such as El Niño, and
Coast strains. Interestingly, the change in V. cholerae strains responsible for pandemic (3) the onset and rapid spread of epidemic
O antigen was accompanied by acquisition cholera. Moreover, the zooplankton feed on cholera by ocean currents along coastal areas.
of a capsule—these changes probably result- phytoplankton and therefore increase markedly zooplankton, p. 290
ing from genes acquired by transduction or in number when warm seas and abundant nutri-

Epidemiology Sewage treatment, handwashing, and chlorinating drinking

The epidemiology of intestinal diseases involves transmission by water are important measures to control diarrheal diseases. In the
the fecal-oral route. A common way this occurs is by ingesting United States, surveillance using PulseNet—a DNA subtyping
food or drinking water contaminated with animal or human feces. resource—helps track illness caused by specific intestinal patho-
Sexual practices that lead to oral-anal contact can also transmit gens. This helps public health agencies detect foodborne outbreaks
intestinal pathogens. so that intervention strategies can be implemented. PulseNet, p. 248
Intestinal pathogens sensitive to acid generally have a high Only a few vaccines are available to prevent diarrheal dis-
infectious dose, because most of the ingested microbes are eases, and these are pathogen-specific. Most are not very effective
destroyed by stomach acid. In contrast, acid-resistant patho- because they fail to elicit a strong enough secretory IgA response
gens have a low infectious dose. Diseases caused by bacteria on the intestinal mucosa to reliably limit pathogen colonization.
that have a low infectious dose are easily transmitted through secretory IgA, p. 362

direct contact as well as in contaminated foods or water. Those

caused by bacteria that have a high infectious dose are generally Cholera
transmitted only in contaminated foods or water. People with Cholera causes potentially fatal diarrhea. Seven cholera pandem-
low gastric acidity are more susceptible to intestinal infections. ics have occurred since the early 1800s, with the seventh one
infectious dose, p. 383
beginning in 1961 in Indonesia and spreading to South Asia, the
Middle East, and parts of Europe and Africa. South America had
Treatment and Prevention remained cholera-free for 100 years until January 1991, when the
Oral rehydration therapy (ORT)—giving appropriate fluids disease abruptly appeared in Peru (see Perspective 24.1). The source
by mouth—can be used to counteract the loss of fluid and elec- of the bacterium was likely a freighter that discharged bilgewater
trolytes (salt) from diarrhea. If the patient drinks water alone, into a Lima harbor. Lima’s water supply was not chlorinated and
however, the intestinal tract cannot absorb enough to keep pace quickly became contaminated. The disease then spread rapidly,
with the amount lost. Fortunately, researchers discovered that so that in 2 years more than 700,000 cases and 6,323 deaths had
glucose increases the absorptive capacity of the intestine. This been reported in South and Central America. The introduction
breakthrough led to the development of what is called oral rehy- of cholera into Haiti in 2010, the first cholera epidemic there in
dration salts (ORS) solution, a highly effective lifesaver in severe over a century, poses a new threat. By mid-2011 there were over
diarrheas regardless of cause. ORS is a mixture of glucose and 300,000 cases and 5,000 deaths reported. pandemic disease, p. 438

various salts (sodium chloride, potassium chloride, and trisodium

citrate) that is commercially available as pre-measured packets to Signs and Symptoms
be dissolved in clean water. The World Health Organization has Cholera is a classic example of severe watery diarrheal disease.
also developed a list of recommended home fluids (RHF) that The diarrheal fluid can amount to 20 liters a day and is described
can be used to prevent dehydration. If oral rehydration cannot be as “rice water stool” because of its appearance. Vomiting also
tolerated—for example, if the patient is vomiting—then intrave- occurs in most people at the onset of the disease, and many people
nous hydration may be required. suffer muscle cramps caused by loss of fluid and electrolytes.
Antibacterial medications are not helpful in most intestinal Severe dehydration can lead to multiple organ failure and death.
infections, and often prolong the illness because they suppress
the normal microbiota. On the other hand, these drugs can be life- Causative Agent
saving in cases where the bacterium invades beyond the intestine. The causative agent of cholera is Vibrio cholerae, a curved,
For children in developing countries, zinc supplements decrease Gram-negative rod. There are several different serotypes, grouped
the length and severity of diarrheal disease. according to differences in their O antigen; O1 is the serotype
 Part IV Infectious Diseases 587

currently circulating. V. cholerae is halotolerant and can grow in Cholera toxin is an A-B toxin. The B (binding) portion
alkaline conditions, two characteristics used to design appropriate attaches irreversibly to specific receptors on the microvilli of the
selective media. O antigen, pp. 60, 247 selective media, p. 95 epithelial cells, allowing the A (active) portion to enter the cells. It
causes severe diarrhea by activating a G protein, which normally
Pathogenesis functions as a sophisticated on/off switch that controls activities
Vibrio cholerae cells are killed by acid, so large numbers must be inside the cell in response to external signals. Cholera toxin chem-
ingested before enough survive passage through the stomach to estab- ically modifies the G protein, locking it in the “on” position. This,
lish infection. Once in the small intestine, surviving bacteria adhere to in turn, results in nonstop activity of an enzyme called adenylate
the small intestinal epithelium using pili and other surface proteins. cyclase, which converts ATP to cAMP. The net effect is high lev-
The bacteria multiply on the epithelial cells but do not visibly damage els of cAMP at the cell membrane, a condition that causes the cell
them. However, the bacteria produce cholera toxin, an enterotoxin to continually secrete chloride. The normal turnover of intestinal
(figure 24.11). This toxin activates ion transport channels in the epi- cells eventually removes the toxin. A-B toxins, p. 392
thelial cell membrane, causing chloride and other electrolytes to exit Cholera toxin is encoded by a bacteriophage that infects
the cell. Water follows the electrolytes, resulting in an outpouring V. cholerae, an example of lysogenic conversion. Synthesis of
of fluid and salts into the intestinal lumen. Although the toxin does both the toxin and pili is regulated by the same bacterial gene.
not affect the large intestine, the volume of fluid is too much to be This means that the two factors required for disease production are
absorbed, causing diarrhea that results in severe dehydration. synthesized at the same time. lysogenic conversion, p. 313

V. cholerae
bacterium

1 The A-B toxin’s B subunit attaches

to receptors on cell membrane;
the A subunit enters the cell.
10 μm
A Plasma membrane
of intestinal cell
B

2 The A subunit locks a G protein 4 cAMP activates ion transport

A
in the “active” mode, turning on channels in the membrane
adenylate cyclase. OFF G protein ON causing Cl– and other electrolytes
to pour out of the cell.
ATP
Cl–
Adenylate K+
3 Adenylate cyclase causes the cyclase Na+
conversion of ATP to cAMP.
HCO3–

H2O

cAMP
5 Water follows electrolytes
out of the cell by osmosis.

FIGURE 24.11 Vibrio cholerae Pathogenesis Vibrio cholerae attaches to the small intestine using pili and begins to produce an A-B toxin.
? How would vaccination against the B subunit of cholera toxin help prevent disease?
588 Chapter 24 Digestive System Infections

Epidemiology number of countries other than the United States. One vaccine
Fecally contaminated water is the most common source of cholera consists of a live genetically altered V. cholerae strain, and
infection, although foods such as contaminated crab, oysters, and the other is killed V. cholerae in combination with the purified
vegetables have also been implicated in outbreaks. A person with recombinant B subunit of cholera toxoid. The main features of
cholera can discharge a million or more V. cholerae cells in each cholera are summarized in table 24.5.
milliliter of feces.
Although cholera is relatively common worldwide, the pan- Shigellosis
demic V. cholerae strain has caused relatively few cases in the Shigellosis is found all over the world, most commonly in areas
United States since the early 1900s. The occasional cases along lacking adequate sewage treatment. Reported cases in the United
the Gulf of Mexico—traced to eating coastal marsh crabs and States average about 21,500 per year, but the true prevalence is
oysters—involved a different strain. much likely higher, because diarrhea often goes unreported.
Ominously, a new V. cholerae strain appeared in India in
1992 and then spread rapidly across South Asia. It belongs to sero- Signs and Symptoms
group group O139 and infects even those people with immunity to
Shigellosis classically involves dysentery, but some Shigella spe-
the seventh pandemic strain. This new strain and its rapid spread
cies cause watery diarrhea. Other signs and symptoms include
suggested it could initiate another pandemic, but fortunately it
headache, vomiting, fever, stiff neck, convulsions, and joint pain.
declined in incidence and remained endemic, causing only resur-
The disease is often fatal for infants in developing countries.
gent localized epidemics.
dysentery, p. 585

Treatment and Prevention

Treatment of cholera depends on the rapid replacement of fluid Causative Agent
and electrolytes before irreversible damage to vital organs can Shigellosis is caused by the four species of Shigella—S. dysenteriae,
occur. If patients are vomiting or are too weak to drink oral rehy- S. flexneri, S. boydii, and S. sonnei. Of these, S. dysenteriae is the
dration solutions, intravenous lines are started. The prompt admin- most virulent, and S. sonnei the least. Shigellosis in developing
istration of intravenous or oral rehydration therapy decreases the countries is typically caused by S. dysenteriae and S. flexneri. In
mortality of cholera from over 30% to less than 1%. the United States, however, S. sonnei causes over two-thirds of
Cholera control depends largely on adequate sanitation the cases. Like other members of the family Enterobacteriaceae,
and safe, clean water supplies. Travelers to areas where chol- Shigella species are Gram-negative rods.
era is occurring are advised to cook food immediately before
eating it. Crabs should be cooked for no less than 10 minutes. Pathogenesis
No fruit should be eaten unless peeled personally by the trav- Shigella species invade intestinal epithelial cells, causing a strong
eler, and ice should be avoided unless known to be made from inflammatory response. To initiate invasion, the bacteria take
boiled water. Orally administered vaccines are available in a advantage of the antigen sampling function of M cells, which

TABLE 24.5 Cholera

1 Vibrio cholerae, the causative Signs and Abrupt onset of massive diarrhea,
bacterium, enters the mouth symptoms vomiting, muscle cramps
with fecally contaminated Incubation period Short, generally 12 to 48 hours
food or drink.
1 Causative agent Vibrio cholerae, an alkali- and salt-
2 The bacteria attach to tolerant, curved Gram-negative rod
epithelial cells of the small
intestine. Pathogenesis Cholera toxin causes chloride, other
electrolytes, and water to pour out of
3 Cholera toxin enters the the intestinal epithelium and into the
cells and causes them to 5 lumen; leads to dehydration and shock.
continuously secrete chloride
ions. Other electrolytes and Epidemiology Ingestion of fecally contaminated food
water follow. or water; sometimes associated with
marine crustaceans
4 The outpouring of water and
electrolytes into the intestinal Treatment Replacement of fluid and electrolytes
lumen causes watery diarrhea. and prevention using intravenous or oral rehydration
4 therapy. Prevented by purifying water
5 Fluid loss causes severe 2 and handwashing; vaccination is
dehydration, resulting in shock 3
available in some countries.
and death unless the fluid can
be replaced.
6 The bacteria exit the body 6
with feces.
 Part IV Infectious Diseases 589

normally transfer microbes to macrophages in Peyer’s patches Although nonmotile, the bacterial cells produce a protein that
(figure  24.12). Once Shigella cells enter the macrophages, they polymerizes host cell actin. This “actin tail” propels the bacterium
escape from the phagosome and multiply in the macrophages’ within the cell, sometimes with enough force to move it into a
cytoplasm. These hidden bacteria are released when the infected neighboring cell. The overall result of invasion and spread is
macrophages die. M cells, p. 358 the death and sloughing of patches of epithelium. The bare areas
With access to the bases of the intestinal epithelial cells, become intensely inflamed, covered with pus and blood, which
Shigella cells attach to specific receptors and induce the epithelial accounts for the signs and symptoms of dysentery.
cells to take them in (figure 24.12). Once inside, the Shigella cells Some strains of Shigella dysenteriae produce a potent
escape again and multiply in the cytoplasm of the epithelial cells. cytotoxin known as Shiga toxin, a chromosomally encoded A-B
toxin. The toxin enters the bloodstream and then the B portion
binds to endothelial cells that line the small blood vessels, par-
ticularly in the kidneys. This allows the A subunit to then enter
the cells, where it reacts with ribosomes, halting protein synthe-
sis, which leads to cell death. Shiga toxin is important because
it is responsible for hemolytic uremic syndrome (HUS),
an often fatal condition that can follow Shigella dysenteriae
dysentery. In HUS, red blood cells break up in the tiny blood
vessels, resulting in anemia and kidney failure. Symptoms
1
Epithelial M cell of HUS sometimes include paralysis or other signs of ner-
cell
Intestinal lumen
vous system injury. Shiga toxin is also produced by strains of
M cells take up Shigella
cells and transport them
Escherichia coli that cause HUS.
across the epithelium. They
multiply in the macrophages Epidemiology
that ingest them, leading to
death of that host cell. Shigellosis is almost exclusively a disease of humans, transmitted
by the fecal-oral route. Unlike V. cholera, Shigella cells are not
Macrophage Shigella cells easily killed by stomach acid, so the infectious dose is small. As
few as 10 cells can start an infection. Shigellosis spreads readily in
overcrowded populations with poor sanitation, such as in refugee
camps and daycare centers. People who engage in anal intercourse
2 Shigella cells attach to the are also prone to contracting the disease. Fecally contaminated
base of the epithelial cells food and water have caused numerous shigellosis outbreaks.
and induce those cells to
take them in. From there,
they escape the endosome Treatment and Prevention
and multiply in the
cytoplasm.
Antimicrobial medications such as ampicillin and co-trimoxazole
Dead
macrophage (a combination of trimethoprim and a sulfonamide) are useful
against susceptible Shigella strains because they shorten the dura-
tion of symptoms and the time during which the pathogens are
3 Shigella cells cause the discharged in the feces. Many strains, however, are resistant to
host cell actin to polymerize. these common medications. Unfortunately, these strains often have
This forms an “actin tail”
that propels a bacterium
R plasmids that encode resistance to several antibacterial drugs.
within the host cell, some- R plasmids, p. 209
times with enough force The spread of Shigella is controlled by using sanitary mea-
to move it into a
neighboring cell.
sures to avoid fecal contamination of food and water supplies.
Two specimens of feces, collected at least 48 hours after stopping
antimicrobial medicines, must be negative for Shigella before a
person is allowed to return to a daycare center or food-handling
4 Neutrophils
Infected epithelial cells die job. Shigellosis cases can be tracked through PulseNet, making it
and slough off. An intense easier for public health agencies to detect outbreaks. Table 24.6
inflammatory response
leads to bleeding and
describes the main features of shigellosis. PulseNet, p. 248
abscess formation.
Escherichia coli Gastroenteritis
Escherichia coli strains are almost universal residents of the intes-
FIGURE 24.12 Shigella Pathogenesis The photomicrograph tinal tracts of humans and a number of other animals. Although
shows the actin tails (green) that form on intracellular Shigella cells most strains are harmless, certain ones produce specific virulence
(orange) and rapidly push these non-motile bacteria from cell to cell. factors that allow them to cause intestinal disease. Other strains—
? Why does shigellosis cause dysentery, whereas cholera causes with different virulence factors—cause urinary tract infections,
watery diarrhea? septicemia, and meningitis. virulence factors, p. 383
590 Chapter 24 Digestive System Infections

TABLE 24.6 Shigellosis Pathogenesis

Escherichia coli strains that cause intestinal disease can be
Signs and Fever, dysentery, vomiting, headache, stiff grouped into six pathovars (pathogenic varieties), based on their
symptoms neck, convulsions, and joint pain
array of virulence factors (table 24.7):
Incubation 3 to 4 days
period ■ Shiga toxin–producing E. coli (STEC). These strains, also
Causative Four species of Shigella, Gram- referred to as EHEC or enterohemorrhagic E. coli, produce
agent negative, non-motile members of the Shiga toxins, a family of functionally identical toxins that
Enterobacteriaceae includes the toxin of Shigella dysenteriae. In STEC strains,
Pathogenesis Pass through intestinal barrier in M cells; the genes for Shiga toxins are encoded by various related
induce uptake from the base of epithelial prophages, an example of lysogenic conversion. Some STEC
cells; induce “actin tails” to spread.
strains produce multiple types of Shiga toxins, and even make
Invasion and spread leads to death and
sloughing of epithelium; inflammation and other toxins as well. Most of the strains identified in outbreaks
ulcerations of intestinal lining. Some strains belong to a single serotype, O157:H7, but there are important
make Shiga toxin. exceptions. STEC strains typically colonize the large intestine,
Epidemiology Fecal-oral transmission; low infectious dose; where they inject effector proteins that cause attaching and
humans generally the only source effacing (A/E) lesions. The intestinal damage results in diar-
Treatment Medications such as ampicillin and rhea, which becomes bloody (hemorrhagic diarrhea) due to the
and prevention co-trimoxazole shorten duration of action of Shiga toxins on the local blood vessels. About 5–10%
symptoms and pathogen excretion; many of people infected with STEC develop hemolytic uremic syn-
strains have R plasmids. Spread is controlled drome (HUS). The STEC strain that caused the 2011 outbreak
by sanitary measures.
in Europe (serotype O104:H4) is unusual because it does not
produce A/E lesions and has characteristics of the EAEC
pathovar, which will be discussed shortly. Shiga toxin, p. 589
lysogenic conversion, p. 313 A/E lesions, p. 585
Signs and Symptoms
■ Enterotoxigenic E. coli (ETEC). These strains make pili that
The signs, symptoms, and severity of E. coli gastroenteritis
allow them to attach to and colonize the small intestine. In
depend on the infecting strain. Some strains cause watery diarrhea
addition, they secrete one or more enterotoxins, one of which
and others cause dysentery. One group can cause hemolytic ure-
is nearly identical to cholera toxin. The genes for adhesin and
mic syndrome (HUS), marked by anemia due to lysis of red blood
toxin synthesis are on plasmids.
cells and kidney failure. dysentery, p. 585 HUS, p. 589
■ Enteroinvasive E. coli (EIEC). These strains invade the
Causative Agent intestinal epithelium, causing a disease similar to shigellosis.
Escherichia coli is a Gram-negative rod, closely related to ■ Enteropathogenic E. coli (EPEC). These strains produce
Shigella species. Most strains ferment lactose, an easily observ- pili that allow them to colonize the small intestine, where they
able trait that distinguishes them from Shigella species. inject effector proteins that cause A/E lesions.

TABLE 24.7 Characteristics of Diarrhea-Causing Escherichia coli

Designation Characteristic Features Clinical Picture

Diffusely adhering Grows as a diffuse layer in a thick mucus-associated Diarrhea, particularly in children
E. coli (DAEC) biofilm on the intestinal epithelium; produces toxins
Enteroaggregative Grows in bricklike aggregations in a thick mucus- Variable symptoms; nausea, watery diarrhea (both acute
E. coli (EAEC) associated biofilm on the intestinal epithelium; produces and persistent)
toxins
Enterotoxigenic Colonizes the small intestine and produces toxins, one Nausea, vomiting, abdominal cramps, massive watery
E. coli (ETEC) nearly identical to cholera toxin diarrhea leading to dehydration
Enteroinvasive Invades the intestinal epithelium, causing a disease very Fever, cramps, blood and pus in the feces
E. coli (EIEC) similar to shigellosis
Enteropathogenic Colonizes the small intestine; induces changes in actin Fever, vomiting, watery diarrhea containing mucus
E. coli (EPEC) filaments, causing the microvilli to be replaced by
pedestals under the bacterial cells; A/E lesions

Shiga toxin– Same as above, except it colonizes the large intestine Fever, abdominal cramps, bloody diarrhea without pus;
producing E. coli rather than the small intestine and releases Shiga toxin some patients develop hemolytic uremic syndrome; most
(STEC) strains identified in outbreaks are serotype O157:H7
 Part IV Infectious Diseases 591

■ Enteroaggregative E. coli (EAEC). These strains produce EPEC strains are an important cause of chronic diarrhea in
pili that allow them to adhere to the intestinal epithelium. infants. The infection is uncommon in breast-fed infants, probably
There, they grow in characteristic aggregations (“bricklike”) because of protective antibodies in breast milk. Outbreaks have
in a thick mucus-associated biofilm. In addition, they produce occurred in hospital nurseries, so the infectious dose for infants
enterotoxins and cytotoxins, damaging the intestinal cells and is thought to be very low. A variety of animals have been shown
evoking an inflammatory response. to harbor EPEC, but their importance as a source of infection is
■ Diffusely adhering E. coli (DAEC). These strains are similar not known.
to EAEC, but rather than forming aggregations, they grow as EAEC strains cause diarrhea in children, travelers, and AIDS
a diffuse layer. patients. A variety of animals harbor EAEC strains, but it is not
clear that the animal strains cause human disease.
Epidemiology DEAC strains have caused diarrhea outbreaks in children, but
relatively little is known about their epidemiology.
Just as the symptoms and pathogenesis of E. coli gastroenteritis
depend on the infecting strain, so does the epidemiology.
STEC strains can be foodborne, and epidemics have involved Treatment and Prevention
ground beef, unpasteurized milk, apple juice, bean sprouts, and Treatment of E. coli gastroenteritis varies according to the
green leafy vegetables. The initial source of infection is often symptoms and infecting strain, but as with any disease,
untreated cow manure, reflecting the fact that cattle are an impor- fluid lost from vomiting and diarrhea should be replaced.
tant reservoir. The infectious dose of STEC strains is typically Antibacterial medications are no longer routinely used because
very low, so in addition to foodborne transmission, the bacteria are most cases are self-limiting. Patients with STEC infections
easily spread by direct contact. reservoir of infection, p. 438 who were treated with antibiotics showed an overall worse
ETEC strains commonly cause diarrhea in infants in devel- outcome.
oping countries as well as travelers visiting those regions. Their Measures to prevent E. coli gastroenteritis include hand-
infectious dose is relatively high, so they typically do not spread by washing, pasteurization of drinks, and thorough cooking of food.
direct contact. ETEC are species-specific, and those that infect ani- PulseNet helps track infections caused by E. coli O157:H7.
mals are responsible for significant mortality in young livestock. Traveler’s diarrhea can usually be prevented with bismuth prepa-
EIEC strains primarily cause disease in young children in rations (such as Pepto-Bismol). Unfortunately, the widespread use
developing countries. Relative to some E. coli strains, they are not of antibiotics to prevent diarrhea has promoted development of
easily spread by direct contact, indicating that the infectious dose resistant strains. Some features of E. coli gastroenteritis are sum-
is not very low. Humans appear to be the only source of infection. marized in table 24.8.

TABLE 24.8 Escherichia coli Gastroenteritis

1 Pathogenic strain of E. coli Signs and Vomiting and diarrhea; sometimes

enters by the fecal-oral route, symptoms dysentery
either directly from an infected 1 Incubation 2 hours to 6 days
person or with contaminated period
food or beverage.
Causative agent Escherichia coli, certain strains only
2 Most strains colonize the small
intestine and produce watery 4 Pathogenesis Various mechanisms; attachment to
diarrhea. small intestinal cells allows colonization;
some strains produce one or more
3 Others invade the large 4 enterotoxins; some strains invade large
intestine and cause dysentery.
intestinal epithelium; others alter actin
4 Some strains produce Shiga 2 polymerization to cause attachment and
toxin, which is absorbed by 3 effacing lesions, and may produce Shiga
the bloodstream and causes toxin
hemolytic-uremic syndrome
Epidemiology Common in travelers; can be foodborne
with damage to red blood cells
or waterborne; fecal-oral route
and the kidneys.
transmission; some strains have an
5 The bacteria exit the body 5
animal source.
with feces.
Treatment Replacement of fluids and electrolytes.
and prevention Prevention methods include
handwashing; pasteurization of drinks,
thorough cooking of meats. Bismuth
compounds help prevent traveler's
diarrhea.
592 Chapter 24 Digestive System Infections

Salmonella Gastroenteritis well. Other contaminated products—including tomatoes, brewer’s

yeast, alfalfa sprouts, protein supplements, raw eggs, dry milk, and
Salmonella gastroenteritis is caused by Salmonella enterica and
even a red dye used to diagnose intestinal disease—have started
can be acquired from many animal sources. An estimated 1.4 mil-
outbreaks. Children are commonly infected by colonized but
lion cases occur in the United States each year, with 400 deaths.
seemingly healthy pets—particularly lizards, snakes, and turtles—
Large outbreaks are usually due to commercially distributed foods
that shed the bacteria in their feces. The disease has been on the
contaminated by animal feces.
rise, largely due to mass production and distribution of foods.
Signs and Symptoms Treatment and Prevention
Signs and symptoms of Salmonella gastroenteritis include diar- Most people with Salmonella gastroenteritis recover without
rhea (sometimes bloody), abdominal cramps, nausea, vomiting, antimicrobial treatment, which is fortunate because S. enterica
headache, and fever. The disease is often short-lived and mild, but strains have shown increasing plasmid-mediated resistance to
that varies depending on the virulence of the infecting strain and antimicrobial medications. Many isolates of Salmonella serotype
the number of cells ingested. Typhimurium are resistant to five or more drugs. Antibiotic-
resistant strains are often associated with more severe illness and
Causative Agent longer hospital stays. Resistance is likely due to the widespread
Salmonella enterica is a Gram-negative rod and, like Shigella and use of subtherapeutic levels of antibiotics in animal feeds, which is
E. coli, a member of the Enterobacteriaceae. The various Salmonella why several countries have now banned this practice. Antibiotics
strains are subdivided into more than 2,400 serotypes based on dif- are not advised for treating humans except in cases involving tis-
ferences in their somatic (O), flagellar (H), and capsular (K) antigens sue or bloodstream invasion.
(see figure 10.9). Each serotype was once considered a separate Control of Salmonella gastroenteritis depends on sanitary
species and given a distinct name. However, DNA sequence data handling of animal carcasses, pasteurizing or irradiating animal
indicates there are only two species—S. enterica and S. bongori, the products, and testing products for contamination. Surveillance and
latter only rarely isolated from humans. serotypes, p. 247 tracing contaminated sources using PulseNet are also important.
The serotype of a Salmonella strain is significant from Adequate cooking kills Salmonella cells, but the center of cooked
both an epidemiological and a disease standpoint. For instance, food does not always reach the 160° Fahrenheit recommended to
certain serotypes cause enteric fever (discussed next) instead of kill foodborne bacteria. This is especially important when cooking
gastroenteritis. Because of the significance, the serotype is often frozen poultry—the outside may appear “well done,” but the cen-
included with the name—for example, Salmonella enterica sero- ter remains relatively cool, allowing bacteria to survive and infect
type Dublin. For convenience, the name is often shortened, as in the unwary diner. Table 24.9 summarizes the characteristics of
Salmonella Dublin. Note that the serotype is not italicized and Salmonella gastroenteritis.
the first letter is capitalized, which distinguishes it from a species
name. Salmonella serotype Typhimurium and Salmonella sero-
type Enteritidis are most commonly isolated in the United States. Typhoid and Paratyphoid Fevers
Typhoid fever and paratyphoid fever are enteric fevers—systemic
Pathogenesis diseases that originate in the intestine. They are caused by specific
Most Salmonella serotypes are sensitive to acid, so millions of cells
must generally be ingested for enough to survive passage through
the stomach to colonize the intestines. Upon reaching the distal
small intestine (the region where the small intestine ends), the bac- TABLE 24.9 Salmonella Gastroenteritis
terial cells attach to specific receptors on the surface of the epithelial
Signs and Diarrhea, vomiting, headache, abdominal pain,
cells. Contact activates a type III secretion system that transfers symptoms and fever
bacterial effector proteins into the epithelial cell. Within minutes,
Incubation Usually 6 to 72 hours
the epithelial cell takes in the bacterial cells by endocytosis (see period
figure 16.6). The bacteria multiply within a phagosome and are dis-
Causative Salmonella enterica, motile, Gram-negative,
charged from the base of the cell by exocytosis. Some bacterial cells
agent members of the Enterobacteriaceae
escape the phagosome and multiply in the cytoplasm. Macrophages
and neutrophils take up any bacteria that are released, but the mac- Pathogenesis Induce uptake by epithelial cells in the region
between the small and large intestines; bacteria
rophages are often destroyed as a result. The infection, however,
multiply in the phagosome and then are
remains localized. The inflammatory response increases epithelial discharged at the base of the cell; inflammatory
cell fluid secretion, causing diarrhea. effector proteins, p. 387 response increases fluid secretion
Epidemiology Ingestion of food contaminated by animal
Epidemiology feces, especially poultry
Most cases of Salmonella gastroenteritis originate from a non- Treatment Treatment with antimicrobial medication
human animal source. The bacteria sometimes survive for months and prevention is usually not advised. Prevention relies on
in soil and water, so they can be spread in untreated manure. adequate cooking and proper handling of food.
Poultry often carry S. enterica, and eggs can be contaminated as
 Part IV Infectious Diseases 593

serotypes of Salmonella enterica and spread from person to per-

TABLE 24.10 Typhoid and Paratyphoid Fevers
son through fecal-oral transmission. Although rare in the United
States, millions of cases occur in developing countries. Signs and Fever, severe headache, constipation, and
symptoms abdominal pain; sometimes followed by
intestinal rupture, internal bleeding, shock,
Signs and Symptoms and death
Patients suffering from enteric fevers often have a progressively
Incubation period 1 to 3 weeks
increasing fever over a number of days, severe headache, consti-
Causative agent Salmonella serotype Typhi (typhoid
pation, and abdominal pain. In severe cases, this is followed by
fever) and Salmonella serotype Paratyphi
intestinal rupture, internal bleeding, shock, and death. (paratyphoid fever), Gram-negative,
members of the Enterobacteriaceae
Causative Agent Pathogenesis Cross the intestinal epithelium via M cells;
Typhoid fever is caused by Salmonella serotype Typhi, whereas taken up by macrophages and then multiply
paratyphoid fever is caused by Salmonella serotype Paratyphi. within them; bacteria are carried in the
These are systemic diseases, so cases are confirmed by blood bloodstream to locations throughout the
body. The Peyer’s patches are sometimes
culture rather than stool culture. destroyed, leading to rupture of the
intestine and hemorrhage.
Pathogenesis Epidemiology Humans are the only source of infection.
Bacteria that cause enteric fever colonize the intestines, cross the Person-to-person fecal-oral transmission.
mucous membrane via M cells, and then resist killing by macro- Treatment Antibiotics are used for treatment. Attenuated
phages. After multiplying within macrophages, the bacteria are and prevention and inactivated vaccines for typhoid fever; no
carried in the bloodstream to locations throughout the body. The vaccine for paratyphoid fever.
systemic infection causes fever, abscesses, sepsis, and shock,
often with little or no diarrhea. The Peyer’s patches are sometimes
destroyed, leading to rupture of the intestine, hemorrhage, and Campylobacteriosis
death. sepsis, p. 674 Peyer’s patches, p. 358
Campylobacter jejuni was first isolated from a diarrheal stool in
Epidemiology 1972, but it then took 5 years to develop a suitable culture proto-
Humans are the only known host for Salmonella serotypes Typhi col. Once this was widely used, C. jejuni infection was found to
and Paratyphi, so the bacteria are spread from person to person. be a common cause of diarrhea.
Often, this occurs via contaminated food or water.
Signs and Symptoms
Some patients surviving typhoid or paratyphoid fever
remain colonized with the causative agents. The bacteria Campylobacteriosis typically leads to fever, vomiting, diarrhea,
actually reside in the gallbladder, where they multiply free of and abdominal cramps. Dysentery occurs in about half the cases.
competition because most other bacteria are killed or inhib-
ited by concentrated bile. Carriers can shed high numbers of Causative Agent
the bacteria for years. Mary Mallone—“Typhoid Mary”—a Campylobacter jejuni is a curved, Gram-negative rod (figure 
young Irish cook living in New York State in the early 1900s, 24.13). It can be cultivated from feces under microaerophilic
was a notorious Salmonella serotype Typhi carrier. She was
responsible for at least 53 cases of typhoid fever over a 15-year
period. In her day, about 350,000 cases occurred in the United
States each year. The incidence is now low due to improved
sanitation, pasteurization, and public health surveillance
measures.

Treatment and Prevention

Antibiotics are used to treat enteric fever, but some Salmonella
serotype Typhi strains are multidrug-resistant, so susceptibility
testing must be done. Most strains in the United States are suscep-
tible to fluoroquinolones. Surgical removal of the gallbladder and
months of antibiotic therapy are often necessary to rid Salmonella
serotype Typhi carriers of their infection.
An attenuated live oral vaccine for preventing typhoid fever is 5 μm
about 50% to 75% effective. An equally effective injectable vac- FIGURE 24.13 Campylobacter jejuni C. jejuni is a common bac-
cine composed of Salmonella serotype Typhi capsular polysac- terial cause of diarrhea in the United States. Color-enhanced scanning
charide is also available. There is no vaccine against Salmonella electron micrograph.
serotype Paratyphi. Table 24.10 summarizes some of the features ? Why was the incidence of this common disease largely unknown
of enteric fever. attenuated vaccine, p. 423 for decades?
594 Chapter 24 Digestive System Infections

conditions using a selective medium to suppress the other intes-

TABLE 24.11 Campylobacteriosis
tinal organisms. selective media, p. 95
Signs and Diarrhea, fever, abdominal cramps, nausea,
Pathogenesis symptoms vomiting, bloody stools

Once ingested, Campylobacter jejuni passes through the stomach Incubation Usually 3 days (range, 1 to 5 days)
period
and penetrates the epithelial cells of the small and large intestines.
The bacteria multiply within and beneath the epithelium and cause Causative agent Campylobacter jejuni, a curved Gram-
negative, microaerophilic rod
a localized inflammatory reaction. Penetration into the blood-
stream is uncommon. Pathogenesis Low infectious dose. The bacteria multiply
within and beneath the epithelial cells,
A mysterious consequence of C. jejuni infection, Guillain- causing an inflammatory response.
Barré syndrome, occurs in about 0.1% of cases. Up to 40% of Bloodstream invasion is uncommon.
all Guillain-Barré cases are preceded by campylobacteriosis, and Complicated by Guillain-Barré syndrome on
autoimmunity is likely responsible in these cases. The syndrome rare occasions.
begins within about 10 days of the onset of diarrhea, with tingling Epidemiology Large foodborne and waterborne
of the feet followed by progressive paralysis of the legs, arms, and outbreaks originating from poultry and
rest of the body. Most patients require hospitalization, but recover other animals; person-to-person spread
rare
completely; about 5% of patients die despite treatment.
Treatment Treatment with antibacterial medications
and prevention is usually not required. Prevention
Epidemiology relies on adequate cooking and proper
Campylobacteriosis is a leading bacterial diarrheal illness in the handling of food, particularly poultry.
United States, with an estimated 2.4 million cases and 125 deaths Water chlorination and pasteurization of
each year. Most of the deaths occur in the elderly and those with beverages are effective control measures.
AIDS or other immunodeficiencies. Numerous foodborne and
waterborne outbreaks of C. jejuni have been reported, involving as
many as 3,000 people. Most cases, however, are sporadic. disease is particularly a problem in healthcare settings, where
Like Salmonella, C. jejuni lives in the intestines of a variety C. difficile is the most common cause of diarrhea.
of domestic animals, including pets and migratory birds. Poultry is
a common source of infection, and up to 90% of raw poultry prod- Signs and Symptoms
ucts harbor the organism. The infectious dose of C. jejuni is as Clostridium difficile–associated disease (CDAD) ranges widely
low as 500 organisms, so one drop of juice from raw chicken in severity. Some patients experience only mild diarrhea, often
meat can easily contain an infectious dose. Unpasteurized milk accompanied by fever and abdominal pain. In other cases, the dis-
and non-chlorinated surface water have also started epidemics. ease progresses to colitis (inflammation of the colon), sometimes
Despite a low infectious dose, person-to-person spread of C. jejuni with patchy areas on the colon called pseudomembranes, a charac-
is rare. teristic of pseudomembranous colitis (figure 24.14).

Treatment and Prevention Causative Agent

Campylobacter jejuni gastroenteritis is typically self-limiting and Clostridium difficile is a Gram-positive, rod-shaped, endospore-
leaves the patient immune to further infection. Erythromycin or forming obligate anaerobe. The endospores are highly resistant to
azithromycin is used to treat severe cases. common disinfectants and environmental conditions, making the
Campylobacteriosis can be prevented by cooking and han- disease control difficult. C. difficile strains that cause CDAD pro-
dling raw poultry properly to avoid cross-contamination. Chicken duce one or more characteristic cytotoxins, and assays that detect
should be cooked until no longer pink (160° Fahrenheit). Pet the toxins are used to diagnose the disease. Some strains appear
owners should wash their hands after contact with animal feces, to be hypervirulent, and the characteristics of these are still under
and outdoor play areas should be kept free of bird droppings. investigation.
Chlorinating drinking water and pasteurizing beverages are also
important control measures. PulseNet helps track Campylobacter Pathogenesis
isolates. The main features of campylobacteriosis are listed in Two toxins (A and B) appear to be important in CDAD. Both
table 24.11. cross-contamination, p. 441 toxins disrupt the host cell actin, causing lethal effects to the intes-
tinal epithelium. In some cases, they cause pseudomembranes—
composed of dead epithelium, inflammatory cells, and clotted
Clostridium difficile–Associated blood—to form on the intestine.
Disease (CDAD)
Clostridium difficile has been recognized as a cause of antibiotic- Epidemiology
associated diarrhea for many years, but the severity of the symp- CDAD primarily occurs in hospitalized patients on antibiotic ther-
toms and the number of outbreaks has been increasing. The apy, probably because C. difficile is able to grow to high numbers
 Part IV Infectious Diseases 595

used. The use of probiotics in treatment has shown promising

results. probiotics, p. 397
Measures to prevent CDAD include minimizing the use of
antibiotics, and avoiding transmission of C. difficile by hand-
washing, wearing gloves, and keeping surfaces disinfected.
Characteristics of Clostridium difficile–associated disease are
summarized in table 24.12.

MicroByte
Fecal transplants—which allow another person’s microbiota to
repopulate the colon—have been successfully used to treat Clostridium
difficile–associated disease.

MicroAssessment 24.4
Shigella, Salmonella, various E. coli strains, Vibrio cholerae,
and Campylobacter jejuni account for most intestinal bacterial
infections. Pathogenic mechanisms of these bacteria include
Pseudomembrane attachment, enterotoxin or cytotoxin production, cell invasion,
FIGURE 24.14 Pseudomembranous Colitis and destruction of microvilli. Dehydration can be treated with
oral rehydration therapy. Clostridium difficile causes antibiotic-
? How does antibiotic therapy predispose a person to this associated diarrhea.
condition?
10. Explain how Vibrio cholerae causes cholera.
11. How does the epidemiology of Salmonella gastroenteritis differ
only when the normal intestinal microbiota has been disrupted. from that of typhoid fever?
Some patients acquire the microorganism while in the hospital, 12. How would you devise a selective medium for Vibrio cholerae? +
but others carry it as a minor component of their normal micro-
biota. Infectious endospores are shed in the feces. Some cases of
CDAD are community acquired, and these patients generally have
an underlying chronic intestinal condition.
24.5 ■ Viral Diseases of the
Treatment and Prevention Lower Digestive System—
When feasible, the antibiotics that predisposed the patient to CDAD
are stopped. This alone often causes symptoms to disappear. If this
Intestinal Tract
is not an option or it fails, then vancomycin, metronidazole, or Learning Outcome
fidaxomicin (Dificid), a new drug developed to treat CDAD, is
9. Compare and contrast the diseases caused by rotaviruses and
noroviruses.

Clostridium difficile–Associated Viral infections of the lower digestive system are common in all
TABLE 24.12
Disease age groups, resulting in millions of cases each year in the United
Signs and symptoms Variable; mild diarrhea to States alone.
pseudomembranous colitis, a
potentially fatal inflammation of
the colon Rotaviral Gastroenteritis
Incubation period Not known; probably a week or less Most cases of viral gastroenteritis in infants and children are
Causative agent Clostridium difficile, a Gram- caused by rotaviruses. Before a vaccine was available, rotavirus
positive, rod-shaped, endospore- infections in the United States resulted in half a million emergency
forming anaerobe
room or clinic visits, and over 55,000 hospital admissions each
Pathogenesis Toxins disrupt host cell actin, year. Worldwide, more than 500,000 children still die from rota-
causing lethal effects to the
intestinal epithelium.
virus infection each year.
Epidemiology Primarily occurs in hospitalized
patients on antibiotic therapy
Signs and Symptoms
Treatment When feasible, stop antibiotics;
Rotaviral gastroenteritis begins abruptly with vomiting and slight
and prevention otherwise, vancomycin, fever, followed in a short time by profuse, watery diarrhea. Signs
metronidazole, or Dificid. and symptoms are generally gone in about a week, but fatal dehy-
dration can occur if fluids are not replaced.
596 Chapter 24 Digestive System Infections

Handwashing, disinfectant use, and other sanitary measures

help limit the spread of rotaviruses. Attenuated vaccines approved
since 2006, administered to infants, are resulting in a substantial
decline in this disease.

Norovirus Gastroenteritis
Noroviruses are the most common cause of viral gastroenteritis in
the United States, responsible for an estimated 23 million cases
annually. They were originally called “Norwalk viruses,” from
Norwalk, Ohio, the place where they were first implicated in an
epidemic of gastroenteritis. The viruses are designated a category
B bioterrorism agent because they spread easily, with the potential
of causing large demoralizing outbreaks. bioterrorism agents, p. 453

Signs and Symptoms

Norovirus gastroenteritis usually causes abrupt onset of nausea,
vomiting, and watery diarrhea. The incubation period is generally
1 or 2 days. Vomiting is typically most severe in older children
FIGURE 24.15 Rotavirus
Color-enhanced electron and adults, and generally resolves within the first day or two.
micrograph. Other symptoms take several days to subside.
? What age group is most commonly infected with rotavirus?
Causative Agent
Noroviruses are naked, single-stranded RNA viruses (figure 24.16).
These viruses represent a group of gastroenteritis-producing
Causative Agent viruses within the calicivirus family, none of which has been cul-
Rotaviruses are naked viruses with a double-walled capsid, and tivated in the laboratory. calicivirus family, p. 308
a double-stranded, segmented, RNA genome (figure  24.15).
The viruses represent a major subgroup of the reovirus family. Pathogenesis
reovirus family, p. 308 Noroviruses infect the epithelium of the upper small intestine,
causing epithelial cell death and decreased production of digestive
Pathogenesis enzymes. The epithelium generally recovers fully within about
Rotaviruses mainly infect the epithelial cells that line the upper 2 weeks. For reasons that are
part of the small intestine. Infection causes epithelial cell death and not understood, immu-
decreased production of digestive enzymes. The damaged lining nity to noroviruses is
fails to absorb fluids, leading to watery diarrhea. In addition, one only short term, lasting
of the viral proteins appears to function as an enterotoxin, causing just several months.
fluid secretion in a manner somewhat similar to cholera toxin. Because of this,
individuals can
Epidemiology have repeated
infections.
Rotaviruses are transmitted by the fecal-oral route. Childhood epi-
demics generally occur in winter in temperate climates, probably
because children are often indoors in groups where the viruses can
spread easily. In regions where vaccination is not common, most
children are infected before age 5. The infection results in some
immunity, so the second and third rotaviral infections cause much
milder diarrhea than the first. Rotaviruses also cause about 25%
of traveler’s diarrhea cases.
Rotaviruses that infect a wide variety of young wild and
domestic animals do not cause human disease. Experimentally,
however, reassortment of genetic segments can occur with dual
infections. It is not known whether new human pathogenic rotavi-
ruses arise by this mechanism.
FIGURE 24.16 Norovirus These viruses cause relatively brief
Treatment and Prevention bouts of gastroenteritis which sometimes go by the name “stomach
Although there is no direct treatment of rotavirus infection, some flu.” Color-enhanced electron micrograph.
infants and small children are hospitalized and given intravenous ? Why are noroviruses so common in crowded settings such as
fluids to prevent dehydration. cruises and college dormitories?
 Part IV Infectious Diseases 597

Rotavirus and Norovirus 24.6 ■ Viral Diseases of

TABLE 24.13 Compared the Lower Digestive
Causative System—Liver
Agent Rotavirus Norovirus
Characteristics Double-walled Single-stranded Learning Outcome
capsid; double- RNA genome 10. Compare and contrast hepatitis A, B, and C.
stranded
segmented RNA
At least five different viruses can cause hepatitis, an inflamma-
genome
tion of the liver, but three types—A, B, and C—account for most
Incubation period 24 to 48 hours 12 to 48 hours
cases (figure 24.17). The viruses are unrelated to each other,
Typical signs Vomiting, Vomiting, but all damage the liver and cause similar signs and symptoms
and symptoms abdominal cramps, abdominal during an acute infection (table 24.14). The most noticeable sign
diarrhea, lasting cramps, diarrhea,
5 to 8 days lasting 12 to
is jaundice—yellowing of the skin and the whites of the eyes.
60 hours Patients with any form of hepatitis should avoid alcohol, acet-
Prevention Attenuated No vaccine
aminophen, and other chemicals known to damage the liver. Note
vaccine that some other diseases, including yellow fever and infectious
mononucleosis, can also damage the liver.

Hepatitis A
Hepatitis A, formerly called infectious hepatitis, is found around
Epidemiology the world. Introduction of an effective vaccine in 1995 signifi-
Transmission of noroviruses is primarily by the fecal-oral route. cantly lowered the incidence of the disease (figure 24.18).
Vomit contains infectious viral particles, however, so transmis-
sion through aerosols and contaminated surfaces can also occur. Signs and Symptoms
The disease is highly contagious due to the low infectious dose Hepatitis A is an acute illness with no known chronic form or
of the virus—about 10 viral particles. The virions are relatively carrier state. Older children and adults with the disease usually
resistant to destruction, and stable in the environment. develop jaundice, fever, fatigue, clay-colored feces, and vomiting
Norovirus epidemics are common on cruise ships and after an incubation period of about 1 month. Most young chil-
college dormitories. Their easy spread also makes them a dren (less than 6 years old) and many older children (ages 6 to
concern for healthcare facilities. More than 50% of foodborne 14) are asymptomatic. About one in five infected adults requires
disease outbreaks are due to norovirus, so the CDC has started hospitalization. Patients generally recover within 2 months, but
CaliciNet, a national surveillance network to trace norovirus some take up to 6 months.
strains.

Treatment and Prevention

35
There are no proven anti-noroviral medications. There is no vac- Hepatitis A
Hepatitis B
cine, and natural immunity to the viruses is short-lived. Thorough Hepatitis C
30
handwashing using soap and water is an important preventative
Cases per 100,000 population

measure. Disinfectants and other sanitary measures also minimize

25
viral transmission. Infected foodworkers should be restricted from
1982—Hepatitis B 1995—Hepatitis A
working for 72 hours after their symptoms subside. Table 24.13 vaccine licensed vaccine licensed
20
compares noroviruses and rotaviruses.

15
MicroAssessment 24.5
10
Rotaviruses are the leading cause of viral gastroenteritis in infants
and children, and they also are a common cause of traveler’s diarrhea.
Noroviruses are the most common cause of viral gastroenteritis. They 5
are highly infectious and easily spread.
13. Why are rotaviral infections common in young children but not 0
adults? 1975 1980 1985 1990 1995 2000 2005
14. Why is handwashing an important means to control the spread of Year
norovirus?
FIGURE 24.17 Incidence of Viral Hepatitis in the United
15. Why might it be more difficult to develop an effective States, 1975–2008
vaccine against noroviruses than against rotaviruses? +
? What organ is affected by hepatitis?
598 Chapter 24 Digestive System Infections

TABLE 24.14 Viral Hepatitis

Hepatitis A Hepatitis B Hepatitis C

Causative agent Naked, single-stranded RNA Enveloped, double-stranded DNA Enveloped, single-stranded RNA
picornavirus, HAV hepadnavirus, HBV flavivirus, HCV
Transmission Fecal-oral Blood, semen Blood, possibly semen
Incubation period 3 to 5 weeks (range, 2 to 7 weeks) 10 to 15 weeks (range, 6 to 23 weeks) 6 to 7 weeks (range,
2 to 24 weeks)
Prevention Inactivated vaccine; immune Subunit vaccine; immune globulin No vaccine
globulin
Comments Usually mild symptoms, but often Acute symptoms often more severe than Usually few or no symptoms;
prolonged; full recovery; no long- hepatitis A; chronic disease can lead to progressive liver damage can lead
term carriers; combined hepatitis cirrhosis and cancer; chronic carriers; can to cirrhosis and cancer; chronic
A and B vaccine available cross the placenta; combined hepatitis A carriers
and B vaccine available

Hepatitis D Hepatitis E

Causative agent Defective single-stranded RNA virus, Naked, single-stranded RNA calicivirus,
HDV HEV
Transmission Blood, semen Fecal-oral
Incubation period 2 to 12 weeks 2 to 6 weeks
Prevention No vaccine No vaccine
Comments Prior or concurrent HBV infection Similar to hepatitis A, except severe
necessary; can cause worsening of disease in pregnant women, same or
hepatitis B; can cross the placenta related virus in rats

Causative Agent known to be damaged by the infection. The virus is released into
Hepatitis A is caused by the hepatitis A virus (HAV), a naked the bile and eliminated with the feces.
single-stranded RNA virus of the picornavirus family. There is
only one serotype of HAV. picornaviruses, p. 308 Epidemiology
Hepatitis A virus spreads by the fecal-oral route, principally via
Pathogenesis fecal-contaminated hands, food, or water. Many outbreaks have
Following ingestion, the virus reaches the liver by an unknown been traced to restaurants where infected food-handlers failed
route. The liver is the main site of replication and the only tissue to wash their hands. Raw shellfish are also a frequent source of
infection because they concentrate the virus from fecally pol-
luted seawater. Groups at high risk of contracting the disease
30 Vaccine available
Midwest
include children in day care centers, residents in nursing homes,
Northeast international travelers, and individuals having sexual contact
25
Cases per 100,000 population

South with an infected person. Because the incubation period averages

West
30 days, HAV can spread widely through a population before
20 being detected. Infected infants and children can shed the virus in
their feces for several months.
15
Treatment and Prevention
10 No antiviral treatment for HAV is available. However, immune
globulin can be given by injection after exposure. This passive
5 immunization gives short-term protection against the disease if
administered within 2 weeks. immune globulin, p. 420
0 An effective vaccine, composed of inactivated HAV, has
1991 1993 1995 1997 1999 2001 2003 2005 2007 been available since 1995. The CDC recommends vaccination
Year for all children 1 year of age, and several high-risk groups such
as people traveling to areas of high incidence or in occupations
FIGURE 24.18 Acute Hepatitis A Reported Cases, different that put them at high risk of exposure. Since the introduction of
regions of the United States, 1991–2007. vaccination, the number of reported cases of hepatitis A has
? What occupations might put a person at risk for hepatitis A? dropped to historic lows. inactivated vaccine, p. 423
 Part IV Infectious Diseases 599

Hepatitis B virions can still be infectious after a week outside the body. HBV
is a member of the hepadnavirus family (hepa-, “liver,” and -dna-
Hepatitis B, formerly known as serum hepatitis, represents about
“DNA”).
half of the cases of viral hepatitis in the United States. Unlike
Three components of HBV are notable because they serve as
HAV, which is passed through the fecal-oral route, the hepatitis
useful markers of infection:
B virus (HBV) is transmitted through contact with body fluids.
■ Hepatitis B surface antigen (HBsAg). This envelope protein
Signs and Symptoms is produced during viral replication in amounts far in excess
of that needed for virion production. The antigen appears in
Signs and symptoms of acute hepatitis B are similar to those of
the bloodstream days or weeks after infection, often long
other forms of hepatitis, ranging from asymptomatic to severe.
before signs of liver damage are evident. Antibodies to
The incubation period varies considerably—from about 2 to
HBsAg confer immunity to HBV.
5 months—depending on the dose received. The acute disease
is rarely fatal, and the virus is usually cleared within weeks to ■ Hepatitis B core antigen (HBcAg). IgM antibodies against
months of initial symptoms. It can become chronic, particularly in this antigen indicate active viral replication.
infants and children. One in five people with a chronic infection ■ Hepatitis B e antigen (HBeAg). High levels of this sol-
develop cirrhosis (scarring of the liver), liver failure, liver cancer, uble component of the viral core correlate with increased
or other chronic liver disease. Between 2,000 and 4,000 people die risk of liver damage and increased risk of spreading the
from HBV-related diseases each year. disease.
The complete hepatitis B virion is also referred to as a Dane
Causative Agent particle. This term provides a contrast to another common viral
Hepatitis B is caused by hepatitis B virus (HBV), an enveloped product of infection—viral envelopes with HBsAg, but lacking
virus that has a mostly double-stranded DNA genome (a portion is DNA (figure 24.19b). These empty envelopes can be 1,000 times
single-stranded) (figure 24.19a). Unlike most enveloped viruses, more common in the bloodstream than Dane particles, which is
HBV is remarkably resistant to environmental conditions—the why HBsAg is a useful marker of infection.

Complete infectious virion Viral envelope particles containing HBsAg

Genomic DNA (double-stranded

with partial single strand)

Hepatitis Envelope
B surface
antigen
(HBsAg)

Nucleocapsid
(viral capsid) Spherical
Envelope lipid
(from host cell)

Hepatitis B e antigen
(HBeAg)
Hepatitis B core antigen
(HBcAg) Elongated

(a) (b)

FIGURE 24.19 Hepatitis B Virus Components Found in the Blood of Infected Individuals (a) Complete infectious virion also known as
a Dane particle. (b) Smaller spherical and elongated envelope particles lacking DNA.
? What does circulating HBeAg indicate about individuals with chronic HBV infection?
600 Chapter 24 Digestive System Infections

Pathogenesis Epidemiology
Following its entry into the body, HBV is carried to the liver by Hepatitis B can be transmitted in body fluids, such as saliva,
the bloodstream. Surface antigen (HBsAg) allows the virus to blood, blood products, and semen. Activities that mix these
attach to and enter host cells. fluids from two individuals are considered risk factors—
The next steps of HBV replication are complex, but they help examples include sharing needles, toothbrushes, razors, or
explain why certain antiviral medications can decrease disease towels, and using unsterile tattooing or ear-piercing instruments.
symptoms. The replication process begins when the viral genome is Unprotected sex is a particularly risky behavior, with nearly half
transported to the host cell nucleus, and the single-stranded gap filled of new hepatitis B cases in the United States acquired through
in (figure 24.20). At this point, the genome is a double-stranded sexual intercourse.
covalently closed circular DNA molecule (cccDNA). A host cell When HBV infection becomes chronic, the virus continues
RNA polymerase uses the cccDNA as a template to produce mRNA replicating—circulating in the blood for many years—even if the
molecules that are then translated to make the various viral proteins. patient is asymptomatic. These infected individuals are extremely
The significant and highly unusual aspect of HBV is that one of the important in the spread of hepatitis B because they are often
RNA molecules serves as a template for DNA synthesis; in other unaware of their infection. The failure to clear the infection is age
words, HBV is a reverse transcribing virus. The RNA molecule— related. More than 90% of infants who contract the virus from an
referred to as pregenomic RNA (pgRNA)—is packaged into the infected mother at or shortly after birth develop a chronic infec-
nucleocapsid along with an HBV-encoded reverse transcriptase. tion. In contrast, 25% to 50% of children infected between the
This enzyme uses the pgRNA as a template to make one strand of ages of 1 and 5, and 6% to 10% of those infected as an older child
DNA; it then uses that DNA molecule as a template to synthesize or adult, will develop a chronic infection.
the complementary strand. The process is not finished inside the A progressive rise in hepatitis B cases was reported in the
virion, however, leaving the HBV genome only partly double- United States from 1965 to the mid 1980s. Since that time, the
stranded. reverse transcribing virus, p. 320 reverse transcriptase, p. 320 HBV vaccine has lowered the incidence of the disease (see fig-
Liver damage from HBV infection is likely due to the cell- ure  24.17). Today, approximately 45,000 new HBV infections
mediated immune response, as effector cytotoxic T cells attack occur each year, and about 1 million people in the United States
infected liver cells. The liver cell destruction leads to cirrho- are chronically infected.
sis. The role of the virus in the development of liver cancer Approximately 2 billion people worldwide have been infected
is not well understood, but in most cases of the cancer, by HBV, with more than 350 million chronic infections. This
the viral DNA has integrated into the host cell genome. “silent disease” is the ninth leading cause of death worldwide.

HBV HBV
8 HBV acquires
its envelope
as it exits from
the cell.
1 HBV infects 7 Reverse transcriptase uses
liver cell. pgRNA as a template to
Liver cell
make DNA but does not
complete the process, leaving
the HBV DNA genome
2 After uncoating, the partially single-stranded.
HBV genome is trans-
ported to the nucleus.

cccDNA 6 Viral particles assemble, and

HBV-encoded reverse transcriptase
is packaged with the pgRNA.
Nucleus pgRNA

mRNA
3 The single-stranded gap in the mRNA
genome is filled in, creating a 5 The mRNA is translated,
covalently closed circular DNA 4 RNA polymerase transcribes Viral proteins producing the virally
molecule (cccDNA). the cccDNA, generating mRNA encoded proteins.
and pregenomic RNA (pgRNA).

FIGURE 24.20 Replication of Hepatitis B Virus

? With respect to the replication strategy, what unusual feature does HBV have in common with HIV?
 Part IV Infectious Diseases 601

Treatment and Prevention Pathogenesis

There is currently no curative antiviral treatment for hepatitis B. Few details are known about the pathogenesis of HCV.
However, some chronically infected patients show remarkable Infection generally occurs from exposure to contaminated
improvement when given reverse transcriptase inhibitors such as blood. The incubation period averages about 6 weeks (range,
lamivudine or adefovir dipivoxil along with injections of geneti- 2 weeks to 6 months). Although most people lack symptoms
cally engineered interferon. reverse transcriptase inhibitors, p.  476 with acute infection, more than 80% develop chronic infec-
interferon, p. 341 tions. The virus infects the liver and triggers inflammatory
The first vaccine against hepatitis B was approved in the and immune responses. After that, the disease process starts
early 1980s. It consisted of HBsAg obtained from the blood of and stops—at times the liver seems to return to normal, then
chronic carriers; 1 ml of their blood often had enough antigen weeks or months later shows marked inflammation. After up
to immunize eight people! Since 1986, however, a subunit to 20 years, cirrhosis and liver cancer develop in 10% to 20%
vaccine produced in genetically engineered yeast has been of patients.
available. It is administered to all infants before they leave the
hospital. A combined vaccine against both hepatitis A and B
Epidemiology
can be used to complete the vaccination schedule. Hepatitis B
vaccination prevents the disease, and may also prevent many Although HCV is transmitted in blood, the mechanism of expo-
of the 500,000 to 1 million new liver cancers that occur world- sure is not always obvious. Approximately 50% of infections
wide each year. in the United States are due to sharing of syringes. Tattoos
Educating groups at high risk of contracting hepatitis B helps and body piercing with unclean instruments have also trans-
prevent the disease. Individuals likely to be exposed to blood— mitted the disease. Other items that can become contaminated
like healthcare workers—are taught to consider all blood infec- with blood and are therefore possible sources of infection
tious and use universal precautions. These precautions include include toothbrushes, razors, and towels. Sexual intercourse
wearing gloves and handling potentially contaminated sharp is an uncommon means of transmission, but individuals with
objects with care (see Perspective 19.1). Teaching the importance multiple partners and other sexually transmitted infections are
and proper use of condoms also helps limit spread of the infection. at increased risk for the disease. The risk of contracting the
disease from blood transfusion is now extremely low because
of effective screening of donated blood.
Hepatitis C
Hepatitis C is the most common chronic, blood-borne infection Treatment and Prevention
in the United States. It was discovered when post-transfusion A combination treatment of interferon and ribavirin sometimes
hepatitis continued to occur regularly even after HBV was prevents chronic disease. In addition, two drugs that inhibit
excluded from transfusions. In 1989, scientists were able to HCV protease have recently been approved for use with the
clone parts of the genome of a transfusion-associated virus, now standard combination.
known as hepatitis C virus (HCV). One of the gene products of No vaccine is available for preventing hepatitis C. However,
these first isolates was HCV antigen, which was then used to physicians often recommend vaccination against hepatitis A and
detect anti-HCV antibody in the blood of prospective donors. B to help prevent a combination of infections that might severely
By eliminating donated blood that contained antibodies against damage the liver.
HCV, the incidence of post-transfusion hepatitis fell dramati-
cally. The antibody test has revealed that more than 3 million
Americans are infected with HCV, and approximately 20,000 MicroAssessment 24.6
new cases occur each year.
Hepatitis viruses are a diverse, unrelated group that cause liver
inflammation. Most cases of hepatitis are caused by hepatitis viruses
Signs and Symptoms A, B, or C. Hepatitis A is transmitted by the fecal-oral route and is
The symptoms of hepatitis C are similar to those of hepatitis A preventable by immune globulin and an inactivated vaccine. Hepatitis
and B except they are generally milder. About 65% of infected B, transmitted by exposure to blood and by sexual intercourse, is
individuals have no symptoms relating to the acute infection, preventable by a subunit vaccine. A combination hepatitis A and
whereas only about 25% have jaundice. In many cases, the infec- hepatitis B vaccine is available. Hepatitis C is transmitted by blood
and occasionally by sexual intercourse. Chronic hepatitis often results
tions become chronic. in cirrhosis and liver cancer.
16. What two serious complications can occur late in the course of
Causative Agent
both chronic hepatitis B and C?
HCV is an enveloped, single-stranded RNA virus of the flavivi-
17. At what stage in the replication of hepatitis B does a reverse
rus family, first cultivated in vitro in 2005. There is considerable transcriptase inhibitor act?
genetic variability, and the variants are classified into types and
18. Why is it possible to immunize multiple people from the blood
subtypes that differ in pathogenicity as well as response to treat- of one hepatitis B patient? +
ment. flavivirus family, p. 309
602 Chapter 24 Digestive System Infections

24.7 ■ Protozoan Diseases of the

Lower Digestive System
Learning Outcome
11. Compare and contrast giardiasis, cryptosporidiosis,
cyclosporiasis, and amebiasis.

Protozoa—single-celled eukaryotes—are important causes of

human intestinal disease. Intestinal protozoan pathogens are all
transmitted by the fecal-oral route. protozoa, p. 291

Giardiasis
Giardiasis is the most commonly identified waterborne illness
in the United States. It can be contracted from clear mountain
streams, chlorinated city water that has not been filtered, and
person-to-person contact. The disease has worldwide distribution
and is responsible for many cases of traveler’s diarrhea.
FIGURE 24.21 Giardia lamblia
Signs and Symptoms
? Does this image show a cyst or a trophozoite?
In epidemics of giardiasis, about two-thirds of exposed individu-
als develop symptoms. The incubation period is generally 6 to 20
days. Symptoms can range from mild (indigestion, “gas,” and nau-
sea) to severe (vomiting, explosive diarrhea, abdominal cramps, intestinal impairment is probably due to the host immune system
fatigue, and weight loss). The symptoms usually end without attacking the parasites.
treatment in 1 to 4 weeks, but some cases become chronic. Both When trophozoites detach from the epithelium, they are car-
symptomatic and asymptomatic persons can become long-term ried by intestinal contents toward the large intestine. If the transit
carriers, unknowingly excreting infectious cysts with their feces. time is long enough, they develop into cysts. Thus, a person who
has formed stools is more likely to excrete cysts, whereas a person
Causative Agent with diarrhea is apt to excrete trophozoites (figure 24.22).
Giardia lamblia is a flagellated protozoan shaped like a pear cut
lengthwise. It has two side-by-side nuclei that resemble eyes and Epidemiology
an adhesive disc on its undersurface that together give the organ- Giardia lamblia is easily spread by the fecal-oral route, because
ism a distinctive appearance (figure  24.21). It can exist in two only 10 cysts are required to establish infection. Water contami-
forms: a growing, feeding trophozoite and a dormant cyst. The nated with human feces is a common source of infection, but feces
cysts have thick walls composed of a tough, flexible, chitinlike from animals such as beavers, raccoons, muskrats, dogs, and cats
polysaccharide that protects the organism from harsh environmen- can also be implicated. The cysts are infectious and can remain
tal conditions. chitin, p. 32 viable in cold water for more than 2 months. Hikers who drink
Because G. lamblia lacks mitochondria, it was thought to from streams, even in remote areas thought to contain safe water,
have evolved before eukaryotes acquired these organelles. More are at risk of contracting giardiasis.
recent evidence appears to refute this idea because the protozoan Although waterborne outbreaks are the most common,
contains atypical energy-metabolizing structures called mito- person-to-person contact can also transmit the disease. This is
somes that likely evolved from mitochondria. especially likely in daycare centers where workers’ hands become
contaminated while changing diapers. Sexual practices that lead
Pathogenesis to oral-anal contact can also transmit giardiasis. Transmission by
The cysts of G. lamblia are infectious because, unlike the tro- fecally contaminated food has also been reported. Good personal
phozoites, they are resistant to stomach acid. From each cyst that hygiene, especially handwashing, decreases the chance of passing
reaches the upper part of the small intestine, two trophozoites on the infection.
emerge. Some of these attach to the epithelium by their adhe- MicroByte
sive disc, whereas others use their flagella to move freely in the A single human stool can contain 300 million G. lamblia cysts.
intestinal mucus. Some may even migrate up the bile duct to the
gallbladder and cause cramping or jaundice.
The trophozoites interfere with the intestine’s ability to Treatment and Prevention
absorb nutrients and secrete digestive enzymes. The result is bulky Several medicines can be used to treat giardiasis, including
feces containing fat, excessive intestinal gas from bacterial diges- tinidazole, metronidazole (Flagyl), and the newest option—
tion of unabsorbed food material, and malnutrition. Some of the nitazoxanide.
 Part IV Infectious Diseases 603

1 Cysts enter the mouth.

3 Trophozoites are 1 μm
released from cysts.

2 Cysts pass through

the stomach to the
4 Trophozoites multiply
lower small intestine.
in intestine.

5 Dehydration in large
intestine stimulates
cyst development.

Trophozoite

Cyst
6 Mature cysts or trophozoites
are eliminated in the feces to
contaminate soil, water, hands,
and food.

FIGURE 24.22 Life Cycle of Giardia lamblia

? Why is an infected person with diarrhea less likely to transmit giardiasis than one who has formed stools?

The level of chlorine used to treat municipal water supplies

does not destroy Giardia cysts, so the water is generally filtered TABLE 24.15 Giardiasis
to remove them. For hikers, the best way to make drinking water Signs and Mild: indigestion, flatulence (intestinal gas), nausea;
safe from giardiasis is to boil it for 1 minute or use a portable filter symptoms severe: vomiting, diarrhea, abdominal cramps,
of 1 μm pore size or smaller. Other methods, including adding weight loss
commercial water-purifying tablets, tincture of iodine, or house- Incubation 6 to 20 days
hold bleach are time-consuming and much less reliable. As in all period
chemical microbial control procedures, time and temperature are Causative Giardia lamblia, a flagellated pear-shaped
important. Only an hour may be necessary to treat warm water, but agent protozoan with two nuclei
many hours are required to kill cysts in cold water. Table  24.15 Pathogenesis Ingested cysts survive stomach passage;
describes the main features of giardiasis. trophozoites emerge from the cysts in the small
intestine, where some attach to epithelium and
others move freely; mucosal function is impaired
by adherent protozoa and host immune response.
Cryptosporidiosis (“Crypto”)
Epidemiology Ingestion of fecally contaminated water; low
Cryptosporidiosis, commonly called “crypto,” was first identi- infectious dose; person-to-person spread
fied as a threat to humans when the AIDS epidemic struck in
Treatment Several treatment options including tinidazole,
the early 1980s. We now know that the disease is a hazard not and metronidazole (Flagyl), and nitazoxanide.
only to those with immunodeficiency but to the public at large. prevention Prevention: boiling, filtering, or disinfecting
A 1993 waterborne outbreak in Milwaukee affected more than drinking water.
400,000 people!
604 Chapter 24 Digestive System Infections

Signs and Symptoms Giardia cysts. They are too small to be removed from drinking
Signs and symptoms of cryptosporidiosis include fever, loss of water by some filtration methods.
appetite, nausea, abdominal cramps, and profuse watery diarrhea. C. parvum is particularly difficult to control because it has a
These begin after an incubation period of 4 to 12 days. The symp- wide host range, infecting domestic animals such as dogs, pigs,
toms generally last 10 to 14 days, but in people with immunode- and cattle. Feces from these animals, as well as from humans,
ficiency diseases, they can last for months and be life-threatening. can contaminate food and drinking water. Epidemics have arisen
from drinking water, swimming pools, a water slide, a zoo foun-
Causative Agent tain, daycare centers, unpasteurized apple juice, and other food
and drink.
Cryptosporidiosis is caused by Cryptosporidium parvum, an api-
complexan. The organism multiplies intracellularly in the small Treatment and Prevention
intestinal epithelium. Unlike other Apicomplexa, its entire life
A new medication has recently been approved to treat
cycle occurs in a single host. The oocyst stage is an acid-fast sphere
cryptosporidiosis—nitazoxanide.
that contains four banana-shaped sporozoites (figure  24.23).
Effective measures for preventing the disease include sani-
Apicomplexa, p. 292 acid-fast, p. 48
tary disposal of human and animal feces and treating water using
filtration, ultraviolet radiation, or ozone. Pasteurizing liquids for
Pathogenesis
consumption is also an important control measures. All food-
The digestive fluids of the small intestine release sporozoites handlers should wash their hands with soap and water regularly,
from ingested oocysts. The sporozoites then invade the epithelial and those with diarrhea should not handle food until symptom-
cells of the small intestine, altering the epithelium and intestinal free. Immunodeficient individuals are advised to avoid contact
villi and causing inflammation. Water and electrolyte secretion with animals, boil or filter drinking water using a 1 μm or smaller
increases, and nutrient absorption decreases. Cell-meditated pore size filter, and avoid recreational water activities. The main
immunity is important in controlling the infection. features of cryptosporidiosis are shown in table 24.16.
Epidemiology
Oocysts of Cryptosporidium parvum passed in feces are immedi- Cyclosporiasis
ately infectious. The infectious dose is as few as 10, so person-to- Cyclosporiasis first came to medical attention in the late 1980s, with
person spread readily occurs with poor sanitation. The organism widely scattered epidemics of severe diarrhea. For several years,
is responsible for many cases of traveler’s diarrhea.
Infected individuals often have prolonged diarrhea and can
continue to eliminate infectious oocysts for 2 weeks or more after TABLE 24.16 Cryptosporidiosis
the diarrhea ceases. These oocysts can survive for up to 6 months
Signs and Fever, loss of appetite, nausea, crampy
in food and water, and are even more resistant to chlorine than symptoms abdominal pain, watery diarrhea
Incubation Usually about 6 days (range, 4 to 12 days)
period
Causative agent Cryptosporidium parvum, an apicomplexan.
Its life cycle takes place entirely within the
epithelial cells of the small intestine.
Pathogenesis Following ingestion of oocysts, sporozoites
are released in the intestine. The
sporozoites invade the epithelial cells,
causing an inflammatory response.
Secretion of water and electrolytes
increases, and absorption of nutrients
decreases.
Epidemiology Oocysts of C. parvum are immediately
infectious. The infectious dose is low, so
person-to-person spread occurs easily.
Infected individuals can discharge the
oocysts for weeks after symptoms subside.
The oocysts resist chlorination, and pass
through many municipal water filtration
10 μm
systems. Wide host range.
FIGURE 24.23 Oocysts of Cryptosporidium parvum Acid-fast Treatment Treated with nitazoxanide. Prevented by
stain of feces. Person-to-person and waterborne transmission are and prevention pasteurization of beverages, boiling or
common. filtering drinking water. Sanitary disposal
? Why is person-to-person transmission more likely with of human and animal feces.
Cryptosporidium infection than with Cyclospora cayetanensis?
 Part IV Infectious Diseases 605

the causative organism was known only as a “cyanobacterium-like TABLE 24.17 Cyclosporiasis
body.” Later, the bodies were shown to be the oocysts of a proto-
zoan, Cyclospora cayetanensis. cyanobacteria, p. 261 Signs and Fatigue, loss of appetite, vomiting, watery
symptoms diarrhea, and weight loss. Symptoms
improve in 3 to 4 days, but relapses can
Signs and Symptoms
occur for up to a month.
Cyclosporiasis begins after an incubation period of about 1 week, Incubation Usually about 1 week (range, 1 to 12 days)
with fatigue, loss of appetite, slight fever, vomiting, and watery period
diarrhea, followed by weight loss. The diarrhea usually subsides Causative agent Cyclospora cayetanensis, an apicomplexan
in 3 to 4 days, but relapses occur for up to 4 weeks.
Pathogenesis Little is known. Biopsies show sexual and
asexual stages in the intestinal epithelium.
Causative Agent
Epidemiology The oocysts of C. cayetanensis are not
Cyclospora cayetanensis is an apicomplexan, and its oocysts are infectious when discharged in the feces,
similar to those of Cryptosporidium parvum but larger. However, and so person-to-person spread does not
C. cayetanensis oocysts are not yet infectious when passed in the occur. Travelers to tropical countries are
feces. With favorable conditions outside the body, sporocysts con- at risk of infection. Produce, especially
raspberries, imported from tropical Central
taining sporozoites develop within the infectious oocyst.
America, has been implicated in most North
American outbreaks.
Pathogenesis
Treatment Treated with co-trimoxazole (trade names
Little is known about the pathogenesis of Cyclospora cayetanensis and prevention Bactrim, Septra). Prevention: Use of boiled
because there is no good animal model for the disease. Analysis of or filtered drinking water is advised in the
small intestinal biopsies of infected patients confirms that sexual tropics. Thorough washing of imported
berries and leafy vegetables.
and asexual forms of the protozoan are both present in the intes-
tinal epithelium.

Epidemiology
The oocysts of Cyclospora cayetanensis are immature and not
however, lasting months or years. Acute dysentery occurs in the
infectious when eliminated in the stool, so person-to-person
most severe cases, which can be fatal.
spread does not occur. In temperate regions, most infections
happen in spring and summer months and among travelers to
Causative Agent
tropical areas. This is likely because warm, moist conditions
favor maturation of the oocysts. Fresh produce (raspberries) has Entamoeba histolytica forms cysts that have a chitin-containing
been implicated in a number of epidemics. In most instances, the wall. Mature cysts—the infectious form for the next host—have
produce was imported from a tropical region, but the source of the four nuclei (figure 24.24).
contaminating organisms is unknown. C. cayetanensis has been
found in natural waters, but the source could not be determined.
Cyst
Treatment and Prevention
Co-trimoxazole (trimethoprim plus sulfamethoxazole) effectively
treats most cases of cyclosporiasis.
No specific preventive measures are available. People should
boil or filter drinking water and thoroughly wash produce such as
berries and leafy vegetables during an outbreak. The main features
of cyclosporiasis are presented in table 24.17.

Amebiasis
Amebiasis is caused by Entamoeba histolytica. Although usually
a mild disease, it causes about 30,000 deaths per year worldwide,
mostly in developing countries. Life-threatening disease occurs in
these areas because virulent E. histolytica strains spread easily in
crowded, unsanitary living conditions.

Signs and Symptoms FIGURE 24.24 Cysts of Entamoeba histolytica

Patients with Entamoeba histolytica infections are commonly ? Why is the organism transmitted in the cyst form and not as a
asymptomatic. Some patients suffer from chronic, mild diarrhea, trophozoite?
606 Chapter 24 Digestive System Infections

other body tissues can result in amebic abscesses. Due to intesti-

TABLE 24.18 Amebiasis
nal ulceration, the diarrhea is often bloody, and the condition is
Signs and Diarrhea, abdominal pain, blood in feces referred to as amebic dysentery.
symptoms
Incubation 2 days to several months Epidemiology
period Entamoeba histolytica has a low infectious dose, spreads by the
Causative agent Entamoeba histolytica fecal-oral route, and is distributed worldwide. The disease is more
Pathogenesis Ingested cysts excyst, releasing
common in tropical areas where sanitation is poor. In the United
trophozoites; these feed on mucus States, cases occur mainly in poverty-stricken areas, and among
and bacteria in the large intestine; migrant farm workers and men who have sex with men. Humans
cytotoxic enzyme kills intestinal cells. The are the only important reservoir.
trophozoites may penetrate the intestinal
wall, and are sometimes carried to the liver Treatment and Prevention
and other organs, resulting in abscesses.
Medications such as metronidazole and paromomycin are avail-
Epidemiology Ingestion of fecally contaminated food or
able for treatment. Prevention of amebiasis depends on sanitary
water; disease associated with poverty,
migrant workers, and men who have sex measures and avoiding fecal contamination of foods and drinking
with men water. Table 24.18 gives the main features of amebiasis.
Treatment Metronidazole, paromomycin. Prevented
and prevention by good sanitation and personal hygiene. The key features of the diseases covered in this chapter are high-
lighted in the Diseases in Review 24.1 table.

MicroAssessment 24.7
Pathogenesis
Ingested cysts of E. histolytica survive passage through the stom- Giardiasis and cryptosporidiosis are common waterborne diseases that
can also be transmitted person to person. Cyclosporiasis is a similar
ach. The cysts then excyst, releasing trophozoites. Upon reaching disease, but is not transmissible person to person. Amebiasis is caused
the large intestine, these trophozoites begin feeding on mucus and by an ameba that ulcerates the large intestinal epithelium, resulting in
intestinal bacteria. The irritating effect of the trophozoites on the dysentery.
cells causes intestinal cramps and diarrhea. Many strains produce 19. What causes the excessive intestinal gas that characterizes
a cytotoxic enzyme that kills intestinal epithelium on contact. The giardiasis?
organisms may penetrate the lining cells and enter deeper tissues 20. Why is cryptosporidiosis difficult to control?
of the intestinal wall. Sometimes, they penetrate into blood vessels
21. How could the fact that Cryptosporidium parvum is acid-fast
and are carried to the liver or other body organs. Multiplication be used in diagnosis? +
of the organisms and tissue destruction in the intestine and in

Diseases in Review 24.1

Digestive System Diseases

Causative Summary
Disease Agent Comment Table
BACTERIAL INFECTIONS OF THE UPPER DIGESTIVE SYSTEM

Dental caries Streptococcus Cariogenic organisms adhere to teeth and produce acids that damage tooth Table 24.1, p. 579
(tooth decay) mutans surfaces; incidence correlates with sugar consumption and lack of preventive
dental care.
Periodontal Certain groups of Gingivitis is the result of the inflammatory response to plaque and tartar at the Table 24.1, p. 579
disease Gram-negative gum line; chronic periodontitis damages the structures that support the teeth,
anaerobes leading to tooth loss.
Acute necrotizing Treponema sp. with Characterized by painful, bleeding gums, abscessed and broken teeth, and Table 24.1, p. 579
ulcerative gingivi- other anaerobes extremely foul breath; associated with poor oral hygiene, particularly in
tis (ANUG) combination with other stresses.
Helicobacter Helicobacter pylori Causes peptic ulcers; associated with gastric cancers; H. pylori withstands Table 24.2, p. 580
gastritis stomach acid by producing urease.

(continued)
Diseases in Review 24.1 (Continued )
Digestive System Diseases

Causative Summary
Disease Agent Comment Table
VIRAL INFECTIONS OF THE UPPER DIGESTIVE SYSTEM

Herpes simplex Herpes simplex Initial infection typically occurs during childhood; latent virus reactivates, Table 24.3, p. 582
(cold sores) virus causing recurrent cold sores.
Mumps Mumps virus Characterized by painful swelling of the parotid glands; virus enters via the Table 24.4, p. 584
respiratory tract and then spreads to glands; can cause sterility in males;
preventable by vaccination.

BACTERIAL INFECTIONS OF THE LOWER DIGESTIVE SYSTEM

Cholera Vibrio cholerae Classic example of watery diarrhea; V. cholerae colonizes small intestine and Table 24.5, p. 588
produces cholera toxin, causing secretion of water and electrolytes; rehydration
is critical.
Shigellosis Shigella species Classic example of bacterial dysentery; Shigella species invade cells of large Table 24.6, p. 590
intestine; low infectious dose; some species produce Shiga toxin.
Escherichia coli Certain strains of Various symptoms depending on the pathovar; STEC strains cause bloody Table 24.7, p. 590;
gastroenteritis E. coli diarrhea and HUS; symptoms of ETEC are similar to cholera; symptoms of EIEC Table 24.8, p. 591
are similar to shigellosis.
Salmonella Salmonella enterica Foodborne, particularly poultry Table 24.9, p. 592
gastroenteritis (not serotypes Typhi
or Paratyphi)
Typhoid and para- Salmonella enterica Salmonella serotypes Typhi and Paratyphi cross intestinal mucosa and invade Table 24.10, p. 593
typhoid fevers serotypes Typhi and bloodstream, causing life-threatening enteric fever; vaccine for typhoid fever is
Paratyphi available.
Campylobacteriosis Campylobacter Foodborne, particularly poultry Table 24.11, p. 594
jejuni
Clostridium Clostridium difficile Typically occurs only in patients on antibiotic therapy; most common cause of Table 24.12, p. 595
difficile–associated diarrhea in healthcare settings.
disease

VIRAL INFECTIONS OF THE LOWER DIGESTIVE SYSTEM—INTESTINAL TRACT

Rotaviral Rotaviruses Most common cause of viral gastroenteritis in infants and children in United Table 24.13, p. 597
gastroenteritis States; preventable by vaccine approved in 2006.
Norovirus Noroviruses Most common cause of viral gastroenteritis in the United States; highly Table 24.13, p. 597
gastroenteritis infectious; epidemics are common on cruise ships and college dormitories.

VIRAL INFECTIONS OF THE LOWER DIGESTIVE SYSTEM—LIVER

Hepatitis A Hepatitis A virus Spreads via fecal-oral route; usually mild symptoms, but often prolonged; Table 24.14, p. 598
(HAV) preventable by vaccination.
Hepatitis B Hepatitis B virus Spreads in blood and semen; reverse transcribing virus; chronic infections can Table 24.14, p. 598
(HBV) result in cirrhosis and liver cancer; chronic carriers; preventable by vaccination.
Hepatitis C Hepatitis C virus Spreads in blood, possibly semen; most common chronic blood-borne infection Table 24.14, p. 598
(HCV) in the United States; chronic infections can result in cirrhosis and liver cancer;
chronic carriers.

PROTOZOAN INFECTIONS OF THE LOWER DIGESTIVE SYSTEM

Giardiasis Giardia lamblia Most commonly identified waterborne illness in the United States; low Table 24.15, p. 603
infectious dose; cysts are chlorine-resistant.
Cryptosporidiosis Cryptosporidium Wide host range; oocysts are chlorine-resistant; low infectious dose. Table 24.16, p. 604
parvum
Cyclosporiasis Cyclospora No person-to-person spread because oocysts must mature in the environment Table 24.17, p. 605
cayetanensis to become infectious; imported produce has been implicated in outbreaks.
Amebiasis Entamoeba Most common in tropical regions where sanitation is poor; low infectious dose; Table 24.18, p. 606
histolytica amebic dysentery results from intestinal damage.
608 Chapter 24 Digestive System Infections

FUTURE CHALLENGES 24.1

Defeating Digestive Tract Diseases
Development of better preventive and treat- and accurate ways of identifying pathogens in 8% increase in clinic visits for diarrhea
ment techniques for digestive tract diseases food, and to utilize newly approved methods with each 1°C increase in temperature
has an urgency arising from massive food and such as meat irradiation. from the normal.
beverage production and distribution methods. Other challenges include: ■ Exploring new prevention and treatment
For example, in 1994, an estimated 224,000 options. Scientists at the University of
people became ill because a tanker truck used ■ Developing additional effective Florida have developed a genetically engi-
to transport ice cream mix had previously car- vaccines against intestinal pathogens. neered Streptococcus mutans that does not
ried liquid eggs. One day’s production from a There is special need for vaccines that produce lactic acid but readily displaces
ground beef factory can yield hundreds of thou- can be administered by mouth and evoke wild strains of the dental decay-causing
sands of pounds of hamburgers, which are soon long-lasting mucosal immunity. bacterium. Researchers at the University
sent to many parts of this or other countries. ■ Exploring the influence of global of Alberta have custom-designed a
The challenge is to better educate the produc- warming on digestive tract diseases. molecule that binds circulating Shiga
ers and transporters, to develop guidelines to A study of Peruvian children by Johns toxin, potentially preventing hemolytic
help them avoid contamination, to develop fast Hopkins University scientists found an uremic syndrome.

Summary
24.1 ■ Anatomy, Physiology, and Ecology Acute Necrotizing Ulcerative Gingivitis (table 24.1)
The digestive system encompasses the digestive tract and accessory Acute necrotizing ulcerative gingivitis (ANUG) can occur at any age in
organs. (figure 24.1) association with poor mouth care and drug abuse (figure 24.5).
The Upper Digestive System Helicobacter pylori Gastritis (table 24.2)
The upper digestive system includes the mouth and salivary glands, Helicobacter pylori predisposes the stomach and the uppermost part of
the esophagus, and the stomach. Bacteria grow on teeth, attached to the duodenum to peptic ulcers (figures 24.6). Treatment with antimicrobial
the pellicle that adheres to the enamel (figure 24.2), forming a biofilm medications can cure the infection and prevent peptic ulcer recurrence.
called dental plaque. Peristalsis in the esophagus propels microbes
to the stomach. The acidity and enzymes in the stomach destroy most 24.3 ■ Viral Diseases of the Upper Digestive System
bacterial cells.
Herpes Simplex (Cold Sores) (table 24.3; figure 24.7)
The Lower Digestive System Herpes simplex is caused by an enveloped DNA virus. HSV persists as
The lower digestive system includes the small and large intestines, as a latent infection inside sensory nerves; active disease occurs when the
well as the pancreas and liver. As the stomach contents enter the small body is stressed.
intestine, digestive fluids from the pancreas and liver are mixed in. Villi
Mumps (table 24.4; figure 24.8)
and microvilli increase the surface area of the intestinal lining. The
large intestine absorbs remaining nutrients and water from the intestinal Mumps is caused by an enveloped RNA virus that infects the parotid
contents and feces are excreted. Microbes make up about one-third glands and a variety of other body tissues. Mumps virus generally
of the weight of feces. The liver produces bile, which is stored in the causes more severe disease in persons beyond the age of puberty; it can
gallbladder. be prevented using an attenuated vaccine (figure 24.9).

UPPER DIGESTIVE SYSTEM INFECTIONS LOWER DIGESTIVE SYSTEM INFECTIONS

24.2 ■ Bacterial Diseases of the Upper 24.4 ■ Bacterial Diseases of the Lower
Digestive System Digestive System
Dental Caries (Tooth Decay) (table 24.1) General Characteristics (figure 24.10)
Plaque that contains Streptococcus mutans or other cariogenic Bacterial infections of the small intestine typically cause watery diarrhea,
microbes can act as a tiny acid-soaked sponge applied closely to the whereas invasion of the large intestine causes dysentery. The bacteria
tooth (figures 24.3, 24.4). Control of dental caries depends mainly on are transmitted via the fecal-oral route, often through contaminated foods
restricting dietary sucrose, supplying fluoride, flossing, and brush- and water. Intestinal pathogens that have a low infectious dose can be
ing teeth. transmitted by direct contact as well. Dehydration can be treated with oral
rehydration therapy (ORT), using oral rehydration salts (ORS).
Periodontal Disease (table 24.1)
Periodontal disease is caused by an inflammatory response to the plaque Cholera (table 24.5)
bacteria at the gum line; it is an important cause of tooth loss in older Cholera is a severe form of watery diarrhea caused by a toxin of
people (figure 24.3). Vibrio cholerae that acts on the small intestinal epithelium (figure 24.11).
 Part IV Infectious Diseases 609

Shigellosis (table 24.6) 24.6 ■ Viral Diseases of the Lower

Shigella species invade the epithelium of the large intestine, causing Digestive System—Liver
dysentery (figure  24.12). Shigella dysenteriae produces Shiga toxin, Hepatitis A (table 24.14; figures 24.17, 24.18)
which causes hemolytic uremic syndrome (HUS).
Hepatitis A virus (HAV) is a picornavirus spread by fecal contamina-
Escherichia coli Gastroenteritis (tables 24.7, 24.8) tion of hands, food, or water. Infection is often asymptomatic in young
Escherichia coli strains that cause intestinal disease can be grouped children; disease in older children and adults is prolonged.
into six pathovars: STEC (or EHEC), ETEC, EIEC, EPEC, EAEC, and Hepatitis B (table 24.14)
DAEC. STEC strains cause hemolytic uremic syndrome.
Hepatitis B virus (HBV) is a hepadnavirus spread by blood, blood
Salmonella Gastroenteritis (table 24.9) products, semen, and from mother to baby (figures 24.19, 24.20). Chronic
Most cases of Salmonella gastroenteritis originate from animals. The infection can lead to cirrhosis and liver cancer.
organisms are often foodborne, commonly in poultry and eggs. Hepatitis C (table 24.14)
Typhoid and Paratyphoid Fevers (table 24.10) Hepatitis C virus (HCV) is a flavivirus transmitted mainly by blood.
Typhoid fever is caused by Salmonella serotype Typhi, and paratyphoid Acute infection is often asymptomatic, and many infections become
fever is caused by Salmonella serotype Paratyphi. Both of these infect chronic, leading to cirrhosis and liver cancer.
only humans. The diseases are characterized by high fever, headache, 24.7 ■ Protozoan Diseases of the Lower
and abdominal pain. Untreated, they can be fatal. Vaccination helps Digestive System
prevent typhoid fever.
Giardiasis (table 24.15)
Campylobacteriosis (table 24.11)
Transmission of Giardia lamblia is usually via drinking water contami-
Campylobacter jejuni is the most common bacterial cause of diar- nated by feces. It is a common cause of traveler’s diarrhea (figures 24.21,
rhea in the United States; it usually originates from domestic animals 24.22). The cysts survive in chlorinated water but can be removed by
(figure 24.13).
filtration.
Clostridium difficile–Associated Disease (CDAD) (table 24.12)
Cryptosporidiosis (“Crypto”) (table 24.16)
Clostridium difficile causes diarrhea and pseudomembranous colitis, The life cycle of Cryptosporidium parvum takes place in the small
primarily in patients on antibiotic therapy (figure 24.14). intestinal epithelium. Its oocysts are infectious, resist chlorination,
and are too small to be removed by many filters (figure 24.23). It is
24.5 ■ Viral Diseases of the Lower Digestive a cause of many water- and foodborne epidemics, and traveler’s
System—Intestinal Tract diarrhea.
Rotaviral Gastroenteritis (table 24.13) Cyclosporiasis (table 24.17)
Rotaviral gastroenteritis is the main diarrheal illness of infants and Transmission of Cyclospora cayetanensis is fecal-oral, via water or
young children but also can involve adults, as in traveler’s diar- produce such as berries; it causes traveler’s diarrhea. Oocysts are not
rhea. Rotaviruses are segmented RNA viruses of the reovirus family infectious when passed in feces, so there is no person-to-person spread;
(figure 24.15). no hosts other than humans are known.
Norovirus Gastroenteritis (table 24.13) Amebiasis (table 24.18)
Norovirus is the most common cause of viral gastroenteritis in the Entamoeba histolytica is an important cause of dysentery; infec-
United States. The viruses are RNA viruses of the calicivirus family tion can spread to the liver and other organs (figure 24.24). Cysts are
(figure 24.16). infectious.

Review Questions
Short Answer Multiple Choice
1. Describe two characteristics of Streptococcus mutans that contrib- 1. Which of the following about intestinal bacteria is false?
ute to its ability to cause dental caries. a) They produce vitamins.
2. Describe the process of periodontal disease. b) They can produce carcinogens.
3. How does Helicobacter pylori cause stomach ulcers? c) They are mostly aerobes.
4. When would a case of mumps likely be complicated by swelling of d) They produce gas from indigestible substances in foods.
the testicles? e) They include potential pathogens.
5. What characterizes the solutions used for oral rehydration 2. All of the following attributes of Streptococcus mutans are impor-
therapy? tant in tooth decay except
6. How do Shigella cells move from one host cell to another even a) it produces endotoxin, which triggers an inflammatory
though they are non-motile? response.
7. Name four different pathogenic groups of Escherichia coli. b) it can grow at pH below 5.
8. What predisposes someone to a Clostridium difficile infection? c) it produces lactic acid.
9. Name two kinds of hepatitis that can be prevented by vaccines. d) it synthesizes glucan.
10. Contrast the cause and epidemiology of giardiasis and amebiasis. e) it stores fermentable polysaccharide.
610 Chapter 24 Digestive System Infections

3. Helicobacter pylori has all of the following characteristics except 9. Which of the following statements about hepatitis B virus is false?
a) it is a helical bacterium with sheathed flagella. a) Replication involves reverse transcriptase.
b) it has not been cultivated in vitro. b) Infected persons may have large numbers of non-infectious
c) it produces a powerful urease. viral particles circulating in their bloodstream.
d) it causes long-term infections, lasting for years. c) In the United States, infection rates have been steadily
e) it can cause stomach ulcers. increasing over the last few years.
4. Vibrio cholerae pathogenesis involves all of the following except d) Asymptomatic infections can last for years.
a) attachment to the small intestinal epithelium. e) Infection can result in cirrhosis.
b) production of cholera toxin. 10. Choose the most accurate statement about cryptosporidiosis.
c) lysogenic conversion. a) Waterborne transmission is unlikely.
d) acid resistance. b) The host range of the causative agent is narrow.
5. Which of the following statements concerning Salmonella enterica c) It is prevented by chlorination of drinking water.
serotype Typhi is false? d) Person-to-person spread does not occur.
a) It is commonly acquired from domestic animals. e) The life cycle of the causative agent occurs within small
b) It can colonize the gallbladder for years. intestinal epithelial cells.
c) It is highly resistant to killing by bile.
Applications
d) It can destroy Peyer’s patches.
1. One reason given by Peruvian officials for not chlorinating their
e) It causes typhoid fever.
water supply is that chlorine can react with substances in water
6. Which statement about rotaviral gastroenteritis is false? or in the intestine to produce carcinogens. How do you assess the
a) A vaccine is available to prevent the disease. relative risks of chlorinating or not chlorinating drinking water?
b) On a worldwide basis, most of the deaths are due to 2. A medical scientist is designing a research program to determine
dehydration. the effectiveness of hepatitis B vaccine in preventing liver cell
c) Most cases of the disease occur in infants and children. cancer. Because liver cell cancer probably has multiple causes,
d) The causative agent infects mainly the stomach. how would you measure the success of an anticancer vaccination
e) The disease is transmitted by the fecal-oral route. program?
7. Which of the following statements about noroviruses is false?
a) They are the most common cause of viral gastroenteritis in the
Critical Thinking +
United States. 1. Why does the lack of a brown color in feces indicate hepatitis?
b) They have a low infectious dose. 2. Mutant strains of Helicobacter pylori that lack the ability to
c) They generally cause vomiting lasting 1 to 2 weeks. produce urease fail to cause infection when they are swallowed.
d) Immunity does not last long. Infection occurs, however, if a tube is used to introduce them
e) They are a category B bioterrorism agent. directly into the layer of mucus that overlies the stomach epithe-
lium. What does this imply about the role of urease in the bacte-
8. Which of the following statements about hepatitis is false?
rium’s pathogenicity?
a) Both RNA and DNA viruses can cause hepatitis.
b) Some kinds of hepatitis can be prevented by vaccines.
c) HCV infections are often associated with injected-drug abuse.
d) Lifelong carriers of hepatitis A are common.
e) Hepatitis A spreads by the fecal-oral route.
25 Genitourinary Tract Infections
KEY TERMS
Catheter A flexible plastic or
rubber tube inserted into the bladder
or other body space, in order to drain
it or deliver medication.
Puerperal Fever Infection
of the uterus following
childbirth, commonly caused by
Streptococcus pyogenes.
Chancre Sore resulting from an Pyelonephritis Infection of the
ulcerating infection. kidneys.
Cystitis Inflammation of the bladder. Toxic Shock Syndrome Collapse
of the blood pressure due to a
Papilloma A kind of tumor
circulating bacterial toxin.
characterized by rough projections of
tissue; warts are an example. Urinary Tract Infection (UTI)
Bacterial infection affecting any part
Pelvic Inflammatory Disease
of the urinary system.
(PID) Infection of the fallopian
tubes, uterus, or ovaries.

difficult to see. Using dark-field microscopy, he saw that the organisms

looked like a corkscrew without a handle—bacteria that are now known
as spirochetes. The spirochetes were found in specimens from other cases
of syphilis, and Schaudinn felt certain he had discovered the cause of
Color-enhanced electron micrograph of a human papillomavirus. syphilis, even though the organism could not be cultured on laboratory
media. Schaudinn named the organisms Spirochaeta pallida, “the pale
spirochete.” This organism is now called Treponema pallidum, from the
Greek words trep and nema, which mean “turning thread.”

A Glimpse of History

 I
nfections of the reproductive and urinary tracts are very com-
Syphilis is an ancient disease that was once very common. History and mon. They are often uncomfortable and can have serious conse-
literature describe many cases of famous people who went insane or were
quences, sometimes without advance symptoms. Urinary tract
seriously disabled because of this disease. A few examples are Henry
infections (UTIs) are the most frequent healthcare-associated
VIII (King of England, 1509–1547), Merriwether Lewis (Lewis and
Clark Expedition, 1803–1808), Oscar Wilde (writer, 1854–1900), and infections and are the main origin of fatal, bacterial blood-
Al Capone (gangster, 1899–1947). stream invasions. Healthcare-associated uterine infections such as
Syphilis was first named the “French pox” or the “Neapolitan dis- puerperal fever, once a common cause of maternal deaths from
ease” because people thought it came from France or Italy. In 1530, the childbirth, still require strict medical attention to be prevented
Italian physician Girolamo Fracastoro wrote a poem about a shepherd (see A Glimpse of History, chapter 19). Sexually transmitted
named Syphilis, who had ulcerating sores all over his body. The descrip- infections (STIs) are widespread and may cause life-threatening
tion matched the signs and symptoms of syphilis, and from then on, the diseases. Although most STIs are transmitted by sexual behavior,
disease was known by the shepherd’s name. some can also be transmitted by drug needles, or through child-
Syphilis was a very severe disease in the early years of the epidemic, birth or breast feeding. puerperal fever, p. 437
and people often died from it, because the available treatments of mercury
The United States has the highest reported incidence of STIs
and natural plant products were not very effective. With time, the disease
among economically developed countries. About 85% of students
became less serious because mutations and natural selection led to more
resistant hosts and a less virulent microbe. have had sexual intercourse, and almost a third of these have
At first it was not known how syphilis was contracted, but gradu- had six or more sexual partners. Approximately 70% of sexually
ally people realized the disease was sexually transmitted. In 1905, the active students report that they or their partners rarely or never use
German biologist Fritz Schaudinn examined fluid from a syphilitic sore. a condom, suggesting a lack of either knowledge or concern about
He observed some motile organisms that were very pale, slender, and STI transmission and risks.

611
612 Chapter 25 Genitourinary Tract Infections

25.1 ■ Anatomy, Physiology, bacteria from ascending to the bladder. The downward flow of
urine also helps clean the system by washing away microorgan-
and Ecology isms before they have a chance to multiply and cause infection.
In addition, normal urine contains organic acids that may make it
Learning Outcome acidic, and antimicrobial substances such as small quantities of
1. Describe the anatomy, physiology, and ecology of the urinary and antibodies. The length and position of the urethra also play a role
genital systems. in preventing infection. Women have a relatively short urethra
(4 cm) emptying close to the anus, and because of this they get
The reproductive and urinary systems are referred to as the UTIs far more frequently than men, who have a longer urethra
genitourinary system because they are positioned close to each (20 cm) more distant from the anus.
other. Both systems are often affected by the same pathogens. If an infection occurs in the urinary tract, an inflammatory
response recruits phagocytes to the bladder, where they engulf
The Urinary System and destroy the invading microorganisms. Lymphocytes in the
infected kidneys or bladder wall also respond, secreting large
The urinary system (or tract) consists of the kidneys, ureters, amounts of antibodies into the urine. phagocytes, p. 346
bladder, and urethra (figure 25.1). The kidneys act as a filtering
system, removing waste materials from the blood and selectively MicroByte
reabsorbing substances that can be reused. Each kidney is con- If urine is alkaline, the person probably has an infection with
nected to the urinary bladder by a ureter. The bladder acts as a a urease-producing bacterium that converts the urea in urine to
holding tank for urine. Once full, it empties through the urethra. ammonia.
Waste materials are excreted in the urine.
The urinary system is protected from infection by a number of
mechanisms. Sphincter muscles near the urethra keep the system The Genital System
closed most of the time and help prevent infections by stopping The female genital system is composed of two ovaries, two
fallopian tubes, a uterus, the vagina and the external genitalia
(vulva), including the labia and clitoris (figure 25.2a). In women
of childbearing age, an egg (ovum) is released from one of the
two ovaries each month during ovulation and is swept into the
Left kidney adjacent fallopian tube. When fertilization occurs, it is normally
Right kidney
in the fallopian tube. Ciliated epithelium of the tube then moves
the fertilized ovum to the uterus, where it implants in the epithelial
lining, developing into an embryo and then a fetus. If fertiliza-
tion does not occur, the epithelial lining of the uterus sloughs off,
producing a menstrual period. The lower end of the uterus is the
Ureters
cervix, which is filled with antimicrobial mucus, except during
menstruation. The cervix opens to the vagina, which leads to the
external genitalia.
Infections of the female genital system can occur in several
Pelvis places. The vagina is a portal of entry for a number of infectious
organisms that can move to the uterus and fallopian tubes during
menstruation. The cervix is a common site of STIs, and is a place
where cancer can develop. Infection of the fallopian tubes can
cause scarring and destruction of the ciliated epithelium, so that
ova are not moved efficiently to the uterus, leading to infertility.
The fallopian tubes are open at both ends, which allows organ-
isms entering the fallopian tubes from the uterus to move into
Bladder the abdominal cavity, where they may infect the liver and other
organs.
Sphincter In men, the reproductive organs include the testes (testicles),
muscles a variety of tubes, ducts and glands, and the penis (figure 25.2b).
Urethra The testes are found outside the abdominal cavity in the scrotum.
Sperm cells from each testis collect in a tightly coiled tubule
FIGURE 25.1 Anatomy of the Urinary System Urine flows called the epididymis and are carried to the prostate gland by a
from the kidneys, down the ureters, and into the bladder, which long tube called the vas deferens. The sperm and secretions of
empties through the urethra. the prostate gland make up the semen. Prostate secretions have
? How does the length and position of the urethra affect the antimicrobial properties. The urinary and reproductive systems
tendency to get urinary tract infections? join at the prostate gland, which can be infected by urinary or
 Part IV Infectious Diseases 613

Bladder

Ureter
Fallopian tube Ureters
Ovary
Pubic
bone
Uterus
Vas Seminal
deferens vesicle
Bladder Prostate
Cervix
gland
Pubic Rectum
bone Urethra
Vagina Anus
Urethra Penis
Epididymis
Anus
Labia Testis

(a) (b)

FIGURE 25.2 Anatomy of the Genital System (a) Female. (b) Male.
? Why can infection of the fallopian tubes result in infertility?

sexually transmitted pathogens. In older men, the prostate often 25.2 ■ Urinary Tract Infections
enlarges and slows the flow of urine, making it more likely that a
UTI will develop. Learning Outcomes
The urine and urinary tract above the urethra are usually
2. Describe the features of bacterial cystitis.
free of microorganisms in both men and women. The lower ure-
3. Outline the characteristics of leptospirosis.
thra, however, has a normal resident microbiota that includes
species of Lactobacillus, Staphylococcus, Corynebacterium,
Urinary tract infections (UTIs) can involve the urethra, bladder, or
Haemophilus, Streptococcus, and Bacteroides. The normal
kidneys, alone or in combination. They account for about 7 million
microbiota of the genital tract of women is affected by the
visits to the doctor’s office each year in the United States. Any
action of estrogen hormones on the epithelial cells of the vagi-
situation that causes inhibition of urination increases the risk of
nal mucosa. When estrogens are present, glycogen is deposited
developing UTIs. Anaesthesia and major surgery, for example,
in these cells. The glycogen is converted to lactic acid by
temporarily stop the reflex ability to urinate, and urine accumu-
lactobacilli, resulting in an acidic pH that inhibits the growth
lates in the bladder. Even being too busy to empty the bladder may
of many potential pathogens. Lactobacilli may also release
predispose a person to infection. Most cases of UTI occur in other-
hydrogen peroxide, an inhibitor of some anaerobic bacteria.
wise healthy young women with normal urinary flow.
Thus, the normal microbiota and resistance to infection of the
female genital tract vary considerably with the person’s hor-
monal status. lactobacilli, p. 258 Bacterial Cystitis (“Bladder Infection”)
Cystitis (inflammation of the bladder) is the most common type
of UTI. Bacterial cystitis is common among otherwise healthy
MicroAssessment 25.1
women, and is also a frequent healthcare-associated infection.
The genitourinary system is one of the portals of entry for pathogens.
The fallopian tubes can provide a passageway for pathogens to enter Signs and Symptoms
the abdominal cavity. The cervix is a common site of infection by
Bacterial cystitis is sometimes asymptomatic, especially among
sexually transmitted pathogens. Prostate enlargement predisposes men
to UTIs. Urine and the urinary tract above the urethra are normally children and the elderly. When symptoms do occur, they typi-
free of microorganisms, but the lower urethra has several resident cally start suddenly and include a burning pain during urination,
genera of bacteria. A woman’s hormones affect vaginal resistance to an urgent need to urinate, and frequent release of small amounts
infection. of urine. The urine is cloudy due to accumulation of leukocytes
1. Describe how the bladder is protected from pathogens. and may be a pale red color due to blood. It also often has a bad
2. Which parts of the genitourinary system are normally sterile? smell. The area above the pubic bone may be painful because of
the underlying inflamed bladder.
3. What changes might occur in the vagina if lactobacilli were
eliminated? + Sometimes a more serious condition called pyelonephritis
occurs. Symptoms of pyelonephritis include fever, chills, vomiting,
614 Chapter 25 Genitourinary Tract Infections

CASE PRESENTATION
A 32-year-old married woman complained numerous rod-shaped, Gram-negative bac- this case. Kidney infections can be detected
of 1 week of burning pain on urination, and teria and neutrophils. Culture of the urine with a scintigram, an image of the kid-
frequent release of small amounts of bloody revealed more than 100,000 colony-forming neys produced following injection into the
urine. About 8 days earlier, she had completed units (CFUs) of Escherichia coli per milliliter. bloodstream of a tiny amount of radioactive
3 days of trimethoprim-sulfamethoxazole The bacterium was resistant to amoxicillin, but material, which is removed from the blood
therapy for similar symptoms. Tests at that sensitive to the other antibacterials useful for by the kidneys and excreted. However, a
time showed that her urine was infected with treating urinary infections. neutrophils, p. 339 scintigram would be dangerous for this
Escherichia coli, resistant only to amoxicil- patient because of her pregnancy.
1. What is the diagnosis?
lin. When the symptoms returned, she began 2. The patient can be treated with
drinking 12 ounces of cranberry juice three 2. What is the treatment?
trimethoprim-sulfamethoxazole, the same
times daily but had only partial relief. She 3. What is the prognosis? medication that was given before. Because
did not have other symptoms such as chills, 4. What future preventive measures would infection in the kidneys takes much longer
fever, back pain, nausea, or vomiting. She was be advisable? to cure than bladder infections, however,
approximately 12 weeks pregnant. the treatment must be continued for 2
Her medical history showed that she had Discussion weeks or longer. Although this drug is
suffered two or three similar episodes of 1. This woman’s signs and symptoms clearly safe to use early in pregnancy, it cannot be
urinary symptoms every year for a number lead to a diagnosis of bacterial cystitis, used late in pregnancy because it can make
of years. Sometimes the symptoms would but there are a number of clues in the jaundice in the newborn worse. A urine
go away when she forced herself to drink presentation of this case that point to a culture was done 1 week after completion
more, but at other times the symptoms would serious complication. First, most patients of treatment to be sure that the infection
persist and she would obtain medical eval- with uncomplicated bacterial cystitis are was truly gone.
uation and treatment with an antibacterial cured by 3 days of an antibacterial medi- 3. The outlook is good for a full recovery
medication. On one occasion several years cation to which the causative bacterium without any permanent damage to the
before the present illness, she had chills, fever, is susceptible. Her signs and symptoms kidneys. The patient was advised, how-
back pain, nausea, and vomiting with her other recurred only 1 day after completing her ever, that repeated future infections of the
urinary symptoms, and she was hospitalized medication. Second, she had signs and kidneys could lead to kidney failure if not
for a “kidney infection.” There was no history symptoms for a full week before she went treated quickly.
suggesting any underlying disease such as dia- for medical evaluation. Third, she was
4. For the future, this patient should use all
betes, cancer, or immunodeficiency. pregnant. Fourth, she gave a past history
the usual methods for preventing UTIs,
The patient was examined and appeared of being hospitalized for pyelonephritis.
including drinking enough to ensure uri-
well, with no obvious discomfort. Her tem- These clues make it highly likely that
nating at least four or five times daily,
perature was normal, as was the rest of her she has a condition called subclinical pyelo-
avoiding delays in emptying the bladder,
physical examination. nephritis, in which her bladder infection has
not using a diaphragm for contraception,
The results of her laboratory tests included spread to her kidneys but has not yet pro-
urinating promptly after intercourse, and
normal leukocyte count and kidney function duced the symptoms of pyelonephritis. As
taking a preventive antimicrobial medica-
tests. Microscopic examination of her urine many as 30% of patients with cystitis have
tion. No vaccine for this infection has been
showed many red and white blood cells. A subclinical pyelonephritis, depending on
approved for use in the United States.
Gram-stained smear of the urine showed risk factors such as the clues mentioned in

back pain, and tenderness overlying the kidneys. Repeated episodes Pathogenesis
of pyelonephritis lead to scarring and shrinkage of the kidneys and The causative agents of cystitis generally reach the bladder by
can cause kidney failure. moving up the urethra, a process helped by motility of the organ-
isms. Uropathogenic E. coli (UPEC) strains that infect the urinary
Causative Agents system have fimbriae that attach specifically to receptors on blad-
Bladder infections usually originate from the normal intestinal der epithelial cells. Bacterial attachment is followed by the death
microbiota. More than 80% of cases are caused by specific uro- and sloughing of this superficial layer of cells. The bacteria then
pathogenic strains of Escherichia coli. The remaining infections enter the underlying epithelium by endocytosis and multiply rap-
in young women are caused by other Enterobacteriaceae mem- idly to create intracellular bacterial communities (IBCs), which
bers such as Gram-negative Klebsiella and Proteus species, or are biofilm-like in nature. Bacteria later detach from the outer
by Gram-positive Staphylococcus saprophyticus. Hospitalized surface of the IBC and move into the bladder lumen to attach
patients, and people with long-standing bladder catheters (tubes to surrounding naive epithelium, creating new IBCs. The UPEC
inserted into the bladder), are often chronically infected with may eventually establish a dormant intracellular reservoir that
multiple species of bacteria, such as Gram-negative Serratia resists antibiotics and is undetected by the immune system, often
marcescens and Pseudomonas aeruginosa and the Gram-positive leading to chronic or recurrent infections. Filamentous forms of
Enterococcus faecalis. Many of these species are resistant to anti- UPEC are observed during IBC development. These forms help
biotics and are difficult to treat. the causative organisms avoid the innate immune response by
 Part IV Infectious Diseases 615

preventing formation of neutrophil-recruiting cytokines and pos-

TABLE 25.1 Bacterial Cystitis
sibly by killing neutrophils already present. Bacteria that move up
the ureters to the kidneys can cause pyelonephritis. fimbriae, p. 65 Signs and Sudden onset, burning pain on urination,
endocytosis, p. 72 symptoms urgency, frequency, foul smell, red-colored
urine; fever, chills, back pain, and vomiting
with pyelonephritis
Epidemiology Incubation period Usually 1 to 3 days
About 30% of women develop cystitis at some time during their Causative agents Most due to Escherichia coli; other
life. Factors that predispose women to UTIs include: Enterobacteriaceae members,
Staphylococcus saprophyticus cause some
■ Short urethra. The length and position of the urethra put it at cases; healthcare-associated infections with
risk for fecal contamination and colonization with potentially antibiotic-resistant strains of Pseudomonas,
pathogenic intestinal bacteria. From the urethra, the bacteria Serratia, and Enterococcus sp.
need only travel a few centimeters to the bladder. Pathogenesis Usually, bacteria ascend the urethra, enter
the bladder, and attach by pili to receptors
■ Sexual intercourse. About one-third of UTIs in sexually
on urinary tract epithelium. Sloughing of
active women are associated with sexual intercourse. The cells and an inflammatory response follow.
massaging effect of sexual intercourse on the urethra moves Spread to the kidneys can occur via the
bacteria from the urethra into the urinary bladder. Many ureters, causing pyelonephritis and possible
women develop bladder infections after having sex for the kidney failure.
first time. Epidemiology Bacterial cystitis is common in women,
promoted by a relatively short urethra, use
■ Birth control devices. Diaphragms compress the urethra and of a diaphragm, and sexual intercourse.
slow the flow of urine, increasing the risk of UTI. Middle-aged men are at risk of infection
because enlargement of the prostate
Other factors involved in development of UTIs include: gland partially obstructs their urethra.
■ Enlarged prostate. UTIs are unusual in men until about Catheterization often results in infection.
age 50, when enlargement of the prostate gland compresses Treatment Treatment: short-term antimicrobial
the urethra and makes it difficult to completely empty the and prevention therapy usually sufficient. Longer
treatment for pyelonephritis. Prevention:
bladder. taking enough liquid to urinate at least
■ Catheterization. Medical conditions may require a bladder four to five times daily, wiping from front
catheter for periods ranging from several days to months. to back. Low daily dose of antibiotic may
help prevent recurrent bacterial cystitis in
This allows bacteria to reach the bladder and establish UTIs.
women.
Pathogens may create a biofilm either within or on the cath-
eter, making it difficult or impossible to kill them with anti-
bacterial medications
■ Paraplegia. Paraplegics (individuals with paralysis of the Cranberry juice and green tea are often used to prevent bacte-
lower half of the body) often have recurrent UTIs. This is rial cystitis, but the scientific evidence on their effectiveness is
because they cannot urinate normally due to lack of bladder inconclusive. The main features of bacterial cystitis are presented
control and require a catheter indefinitely to transfer their in table 25.1.
urine to a container.

MicroByte Leptospirosis
In the United States, about 500,000 hospitalized patients develop Leptospirosis is mainly a disease of animals, but it can be passed
bladder infections each year, mostly after catheterization. to humans. The causative bacterium enters the body through a
mucous membrane or wound and is then carried to the urinary sys-
tem by the bloodstream. The disease is probably one of the most
Treatment and Prevention common of all the zoonoses, but many cases are mild, resolve
Cystitis is usually easily treated with a few days of an anti- without treatment, and remain undiagnosed. More severe cases
microbial medication effective against the causative bacterium. require treatment and can sometimes be fatal. zoonoses, p. 439
Pyelonephritis, a more serious condition, usually requires hospi-
talization and intravenous antibiotic treatment. Signs and Symptoms
General ways to prevent UTIs include drinking enough to Leptospirosis infections are often asymptomatic. When signs and
ensure urinating at least four or five times daily, urinating imme- symptoms do occur, they begin after an incubation period averag-
diately after sexual intercourse, and wiping from front to back ing 10 days. In mild cases, which are most common, symptoms
after defecation to minimize fecal contamination of the urethra. are flulike and include the sudden development of a headache,
Many antimicrobial medications accumulate in the urine, reach- spiking fever, chills, and muscle pain. A dry cough may occur. A
ing concentrations higher than in the blood, so taking a low dose characteristic feature of this first phase (septicemic) is photopho-
of an antibiotic daily can be used to prevent recurrent infections. bia (sensitivity to light) and red eyes, caused by dilation of small
616 Chapter 25 Genitourinary Tract Infections

blood vessels. These symptoms usually fade within a week, and all to be an immune response, and this second phase of the illness is
signs and symptoms of illness are gone in a month or less. therefore called the “immune phase.” This phase is characterized
In severe cases of the disease, symptoms recur after 1 to by injury to the cells that line the lumen of tiny blood vessels,
3 days of feeling well. Symptoms of this second (immune) phase causing clotting and impaired blood flow in tissues throughout
include those of the septicemic phase as well as bleeding from the body, and leading to most of the serious effects of the illness,
various sites, vomiting, rash, and confusion. including kidney failure, the main cause of fatalities.
In rare cases, people with leptospirosis develop Weil’s dis-
ease, which affects the liver and kidneys. Signs and symptoms of Epidemiology
Weil’s disease include jaundice (yellowing of skin and eyes), liver Leptospirosis occurs around the world in all types of climates,
and kidney failure, hemorrhage in many organs, and meningitis. although it is more common in the tropics. Leptospira interrogans
People who suffer biphasic leptospirosis or Weil’s disease need infects many species of wild and domestic animals, usually caus-
medical attention, since these can be fatal. ing little or no apparent illness, but sometimes causing fatal, epi-
demic disease. The organisms are excreted in the animal’s urine,
Causative Agent and urine spots on the ground remain infectious for as long as they
Leptospirosis is caused by Leptospira interrogans, a slender aero- are still moist. Contaminated urine is the main mode of transmis-
bic Gram-negative spirochete with hooked ends (figure  25.3). sion to other hosts. L. interrogans can also survive in mud or water
There are more than 250 antigenic types of this species, some of for several weeks, and there is a correlation between the amount of
which were given different names in the past. Most strains can be rainfall and leptospirosis incidence. Humans contract leptospirosis
cultivated using cell-free media. spirochetes, p. 272 from water, soil, or food contaminated with infected animal urine.
Many cases are caused by swimming in urine-contaminated fresh
Pathogenesis water. Ingested organisms enter through the mucosa of the upper
Leptospirosis interrogans enters the body through mucous mem- digestive system. Infected humans usually excrete the organisms
branes, eyes and breaks in the skin. No lesion develops at the site for a few weeks to several months, but person-to-person transmis-
of entry, but the organisms multiply and spread throughout the sion does not seem to occur.
body by way of the bloodstream, penetrating all tissues including
Treatment and Prevention
the eyes, and the brain. Severe pain is characteristic of this first
(septicemic) phase, and victims sometimes are given unneces- Mild cases of leptospirosis resolve without treatment, although
sary surgery because it is assumed the pain is from appendicitis antibiotics can be given. In biphasic leptospirosis, a number of
or gallbladder infection. There are no inflammatory changes or different antibiotics are effective for treating the disease, but only
tissue damage at this stage. Within a week, the immune response if they are started during the first 4 days of the initial illness. Once
destroys the organisms present in most tissues, although they con- treatment begins, the person often shows a temporary worsening
tinue to multiply in the kidneys. Many people recover at this stage, of symptoms due to massive release of antigens from organisms
with no further development of symptoms. lysed by the antimicrobial medication.
If the disease progresses, the infected person then usually There are few effective measures for preventing leptospiro-
seems healthy for a few days before the second phase of signs and sis other than avoiding animal urine. Multivalent vaccines (that
symptoms occurs. The cause of the recurrent symptoms is thought contain a number of different serotypes of L. interrogans) are
available for preventing the disease in domestic animals, but they
do not always prevent the animal from becoming a carrier. Small
Hook doses of a tetracycline antibiotic can prevent the disease in high-
risk individuals, such as veterinarians, slaughterhouse workers,
farmers, sewer workers, and water sport enthusiasts. The main
features of leptospirosis are shown in table 25.2.

MicroAssessment 25.2
Situations that affect normal urine flow predispose a person to UTIs.
Bladder infections are common, especially in women, and are usually
caused by normal intestinal bacteria ascending from the urethra.
Pyelonephritis is a serious complication that may cause death due
to kidney failure. In leptospirosis, a widespread zoonosis spread by
urine, the causative organisms enter the urinary system from the
bloodstream. Although usually mild, the disease can be a severe
biphasic illness characterized by tissue invasion and pain in the first
phase and tissue destruction and kidney damage in the second.
4. What organism causes the majority of bladder infections in
otherwise healthy women? From where does this organism
5 μm come?
FIGURE 25.3 Leptospira interrogans, the Cause 5. How do people become infected with Leptospira interrogans?
of Leptospirosis
6. How can catheterization lead to bladder infection? +
? What is the main mode of transmission of this organism?
 Part IV Infectious Diseases 617

TABLE 25.2 Leptospirosis

1 Water or animal urine Signs and Many mild and asymptomatic cases.
contaminated with Leptospira symptoms Others have a biphasic illness:
interrogans splashes onto spiking fever, headache, muscle
mucous membrane or abraded pain, bloodshot eyes in the first
1 (septicemic) phase, then 1 to 3 days of
skin.
improvement; heart, brain, liver, and
2 The bacteria infect the
kidney damage in the second (immune)
bloodstream and are carried
phase
throughout all the body tissue
causing fever, intense pain. Incubation period Usually about 10 days (range, 2 to
2 30 days)
3 Signs and symptoms resolve
and bacteria disappear from Causative agent Leptospira interrogans, a spirochete
blood and tissues, except with many serotypes
kidneys. 4 Pathogenesis The bacteria penetrate mucous
3
4 Biphasic illness associated with 4 membranes or breaks in the skin,
severe damage to liver and 5 multiply in the bloodstream, and
kidneys. are carried to all parts of the body.
Septicemic phase: severe pain with
5 Complete recovery occurs
penetration of body tissues, but little
if kidney failure can be
or no tissue damage. Immune phase:
effectively treated.
damage to cells that line small blood
6 Excretion of L. interrogans vessels and clotting of blood. Causes
continues in urine. 6 severe damage to the liver, kidneys,
heart, brain, and other organs.
Epidemiology Worldwide distribution. Wide range
of animal hosts chronically excrete
the bacteria in their urine, causing
contamination of natural waters and
soils. Organisms remain infectious
1 under warm, moist, neutral or alkaline
conditions for long periods of time.
Treatment Treatment: various antibacterial
and prevention medications useful in treatment of
leptospirosis but only if given early in the
disease. Prevention: avoiding contact
with animal urine. Vaccines prevent
disease in domestic animals, may not
prevent urinary carriage. Tetracycline
antibiotics preventive in epidemics.

25.3 ■ Genital System Diseases Signs and Symptoms

BV is characterized by a thin, grayish-white, slightly bubbly vagi-
Learning Outcome nal discharge that has a characteristic strong fishlike smell. The
4. Compare and contrast bacterial vaginosis (BV), vulvovaginal bacteria associated with BV may spread to the uterus or fallopian
candidiasis (VVC), and staphylococcal toxic shock syndrome. tubes, causing pelvic inflammatory disease (PID), which can
lead to sterility. About half of BV cases are asymptomatic.
The genital tract is the portal of entry for many infectious diseases,
both non-sexually and sexually transmitted. This section discusses Causative Agent
some genital system diseases that are not generally transmitted The cause or causes of BV are unknown. Because most cases
sexually. show a significant decrease in vaginal lactobacilli, conditions
that suppress lactobacilli or promote the growth of other micro-
biota are thought to play a causative role. The discharge that
Bacterial Vaginosis (BV) characterizes BV contains large numbers of bacteria, including
In the United States, bacterial vaginosis (BV) is the most common aerotolerant Gardnerella vaginalis, anaerobic species of the gen-
vaginal disease of women in their childbearing years. It is termed era Mobiluncus and Prevotella, Mycoplasma sp., and anaerobic
vaginosis rather than vaginitis because there are no inflammatory streptococci. Although these species are generally present in
changes. In the United States, BV is common in pregnant women, women with BV, women voluntarily inoculated with cultures of
and puts them at risk for premature deliveries. these organisms do not always develop the disease. Also, each
618 Chapter 25 Genitourinary Tract Infections

of these species can occur in vaginal secretions of healthy women,

TABLE 25.3 Bacterial Vaginosis
although typically in much smaller numbers than in women show-
ing symptoms of BV. However, since discharge from women with Signs and Gray-white vaginal discharge and
BV can cause the disease in healthy women, it is suggested that symptoms unpleasant fishy odor
although these species do not necessarily cause BV, they could Incubation period Unknown
play a role in developing the disease. Causative agent Unknown
Pathogenesis Uncertain. Marked change in normal
Pathogenesis microbiota composition. Increased
Women with BV have characteristic changes in the vagina, sloughing of vaginal epithelium in the
absence of inflammation. Odor due to
including a loss of acidity of the vaginal secretions (normally
metabolic products of anaerobic bacteria.
pH 3.8–4.2), disruption of the normal microbiota, and substantial May cause complications of pregnancy,
increase in the numbers of clue cells. Clue cells are epithelial including premature births.
cells that have sloughed off the vaginal wall and are covered with Epidemiology Associated with many sexual partners or a
bacteria (figure  25.4). There is no inflammation unless another, new partner, but can occur in the absence
concurrent vaginal infection is present. The strong fishy odor is of sexual intercourse. No evidence that it is
caused by metabolic products of the anaerobic bacteria and is used a sexually transmitted infection.
for diagnosis in the whiff test. Treatment Treatment with metronidazole is effective.
and prevention No proven preventive measures.
Epidemiology
The epidemiology of BV is not well understood because the caus-
ative agent is not known. The disease is most common among
sexually active women and sometimes occurs in children who important in pregnant women because BV may cause premature
have been sexually abused. Pregnant women are at increased risk birth. Studies on using yogurt vaginally to restore lactobacilli have
of BV. Women who wear thongs, douche, have multiple sex part- given conflicting results.
ners, have sex with other women, have a new sex partner, or use BV can be prevented by abstinence, limiting the number
an intrauterine device (IUD) also have an increased risk. There is of sex partners, and avoiding douching and the use of thongs.
no proof that bacterial vaginosis is sexually transmitted. However, Treatment of the male sex partners of patients with BV does not
women with BV are at higher risk of getting other STIs such as prevent recurrences. The main features of BV are summarized in
gonorrhea, HIV, or chlamydia. Virgins seldom get BV. table 25.3.

Treatment and Prevention

Vulvovaginal Candidiasis (VVC)
Most cases of BV respond quickly to treatment with antibiot-
ics such as metronidazole or clindamycin, which can be given Vulvovaginal candidiasis (VVC), a fungal infection, is the second
orally or vaginally. The disease can recur. Treatment of BV is most common cause of vaginal symptoms after BV. Like BV, it
seems to occur after a disruption of the normal microbiota. As the
name indicates, VVC often involves not only the vagina, but the
vulva as well.

Signs and Symptoms

The most common signs and symptoms of VVC are constant,
intense itching and burning of the vagina or vulva. Typically, there
is a large amount of thick, clumpy whitish or whitish-gray vaginal
discharge. The vaginal mucosa is usually red and somewhat swol-
len, and may have patches of cottage cheese–appearing clumps
attached to it.

Causative Agent
VVC is caused by Candida albicans, a yeast that is part of the
normal microbiota of the vagina in about a third of all women
(figure 25.5). Because C. albicans is a fungus, it has a eukaryotic
cell structure. yeast, p. 283
Clue cell

FIGURE 25.4 Clue Cell in an Individual with Bacterial

Pathogenesis
Vaginosis The cells in the photograph are stained epithelial cells Normally, vaginal colonization by Candida albicans causes no
that have sloughed from the vaginal wall, one of which, the clue symptoms. The growth of the organism is usually limited by the
cell, is a dark color. immune system and the normal vaginal lactobacilli that occupy
? Why does the clue cell appear darker in color than other cells? the same niche and compete for nutrients. When the normal
 Part IV Infectious Diseases 619

Budding
yeast cell TABLE 25.4 Vulvovaginal Candidiasis
Signs and Itching, burning, thick, white vaginal
symptoms discharge, redness and swelling
Incubation period Usually unknown. Generally 3 to 10 days
when associated with antibacterial
medications
Causative agent Candida albicans, a yeast
Pathogenesis Inflammatory response to overgrowth of
the yeast, which is often present among
the normal microbiota
Epidemiology Not contagious. Usually not sexually
transmitted. Associated with antibacterial
therapy, use of oral contraceptives,
pregnancy, and uncontrolled diabetes,
but most cases have no identifiable
predisposing factor.
Pseudomycelium Cell Epithelial 20 μm
Treatment Intravaginal antifungal medications such as
nucleus cell and prevention clotrimazole usually effective. No proven
FIGURE 25.5 Candida albicans Vaginal discharge of a woman preventive measures.
with vulvovaginal candidiasis, showing C. albicans.
? What usually prevents VVC from occurring?

been removed from the market (figure 25.6). The term toxic shock
microbiota balance is disturbed—as occurs during menstruation syndrome was used to describe the signs and symptoms of the ill-
or pregnancy, or when using oral contraceptives or antibiotics— ness. Now that we know its cause, it is called staphylococcal toxic
C. albicans can multiply freely, causing an inflammatory response. shock syndrome. This is not a new disease, but one that emerged in
The signs and symptoms of VVC occur within about 10 days. a new form and became much more common as a result of changes
in technology and human behavior. emerging diseases, p. 448
Epidemiology
Factors that predispose a woman to Candida infection are late Signs and Symptoms
pregnancy, poorly controlled diabetes, and the use of oral contra- Staphylococcal toxic shock syndrome is characterized by the sud-
ceptives or antibiotics. Hormone replacement therapy may also den development of high temperature, headache, muscle aches,
increase the risk of VVC. However, most patients with VVC have bloodshot eyes, vomiting, diarrhea, a sunburnlike rash, and confu-
no known predisposing factors. The disease does not spread from sion. Typically, the skin peels about a week after the development
person to person.

Treatment and Prevention

Intense publicity; suspect Total
Intravaginal treatment of C. albicans infections with antifun- 1,400 tampon taken off the market. Menstrual
gal medicines such as nystatin, clotrimazole, or fluconazole is Non-menstrual
1,200
usually effective. Fluconazole (Diflucan) taken by mouth is Absorbency lowered
generally safe and effective, although it may cause side effects 1,000
Number of cases

such as headache and nausea, and it can interact with other

800 1982–FDA* requires
medications, causing rare, but serious reactions. Self-diagnosis tampon labeling.
and treatment with over-the-counter medications can lead to 600
development of drug-resistant organisms.
Prevention of VVC depends on minimizing the use and 400 FDA standardizes absorbency labeling.
duration of antibacterial medications and on effective treat-
200
ment of underlying conditions such as diabetes. The main
features of VVC are presented in table 25.4. 0
1980 1982 1984 1986 1988 1990 1992 1994 1996 2010
Year
Staphylococcal Toxic *FDA, Food and Drug Administration
Shock Syndrome
FIGURE 25.6 Staphylococcal Toxic Shock Syndrome, United States
Toxic shock syndrome was described in the late 1970s in A sharp drop in cases occurred when a brand of high-absorbency tampon
several children with staphylococcal infections. In 1980, it was taken off the market. Since 1996, only six or fewer total cases have
became epidemic in young, healthy, menstruating women who generally been reported per 100,000 population each year.
were using a brand of high-absorbency tampon that has since ? How does tampon use increase the risk of toxic shock syndrome?
620 Chapter 25 Genitourinary Tract Infections

of the disease. Without treatment, the blood pressure can drop, Staphylococcal Toxic
leading to multiorgan failure, coma, and sometimes death. TABLE 25.5
Shock Syndrome
Causative Agent Signs and Fever, vomiting, diarrhea, muscle aches, low
symptoms blood pressure, and a rash that peels
Staphylococcal toxic shock syndrome is caused by strains of
Staphylococcus aureus that produce toxic shock syndrome Incubation period 3 to 7 days
toxin-1 (TSST-1) or other related exotoxins. Staphylococcus Causative agent Certain toxin-producing strains of
aureus, p. 524 exotoxins, p. 391 Staphylococcus aureus
Pathogenesis Toxin (TSST-1 and others) produced by certain
Pathogenesis strains of S. aureus; toxins are superantigens,
causing cytokine release and drop in blood
Tampon-associated toxic shock syndrome usually begins 2 to 3 pressure.
days after the start of menstruation when tampons are used. The
Epidemiology Associated with certain high-absorbency
staphylococci grow in the blood-soaked tampon. The bacteria tampons, leaving tampons in place for long
rarely spread throughout the body, but as they multiply they periods of time, and abrasion of the vagina
produce TSST-1 or other exotoxins. Staphylococcal toxic shock from tampon use. Also as a result of infection
syndrome results from absorption of these toxins into the blood- by certain toxin-producing S. aureus strains in
stream. The toxins are superantigens that cause activation of large other parts of the body.
numbers of helper T cells, leading a massive release of cytokines Treatment Antimicrobial medication effective against
(cytokine storm). This in turn causes a drop in blood pressure and and prevention the causative S. aureus strain; intravenous
fluids. Awareness of symptoms. Prompt
multiorgan failure—the most dangerous aspect of the potentially treatment of S. aureus infections; frequent
fatal illness. TSST-1 causes 75% of all toxic shock syndrome change of tampons by menstruating women.
cases. superantigen, p. 393

Epidemiology
Staphylococcal toxic shock syndrome can occur after infection
with any strain of Staphylococcus aureus that produces one of the MicroAssessment 25.3
responsible exotoxins. The syndrome can occur after infection of Bacterial vaginosis (BV), the most common vaginal disease of women
surgical wounds, infections associated with childbirth, and other in their childbearing years, is characterized by a significant change
types of staphylococcal infections. It does not spread from person in the composition of the normal vaginal microbiota. Vulvovaginal
to person. Using tampons increases the risk of staphylococcal toxic candidiasis (VVC) often occurs as a result of antibacterial therapy
shock, and the higher-absorbency tampons may pose a greater suppressing normal vaginal microbiota, but many other cases arise
risk. Use of intravaginal contraceptive sponges also increases risk. for unknown reasons. Staphylococcal toxic shock syndrome is caused
by certain strains of Staphylococcus aureus that produce exotoxins,
People who have recovered from the disease are not always immune
which are absorbed into the bloodstream, causing the massive release
to it. Since 1990, there has been a slow, steady decline in the inci- of cytokines responsible for shock.
dence of staphylococcal toxic shock syndrome, now estimated to be
7. What is the causative agent of bacterial vaginosis (BV)?
six or fewer total cases per 100,000 people per year (figure 25.6).
8. Why was the incidence of staphylococcal toxic shock syndrome
higher in menstruating women than in other people during the
Treatment and Prevention early 1980s?
Staphylococcal toxic shock syndrome is a severe disease and 9. Why would using antibiotics predispose a woman to vulvovaginal
requires hospitalization. It can be effectively treated with anti- candidiasis (VVC)? +
bacterial medication active against the infecting S. aureus strain,
intravenous fluid, and other measures to prevent shock and kidney
damage. The source of the infection should be removed if pos-
sible. Although most people recover fully in 2 to 3 weeks, the 25.4 ■ Sexually Transmitted
disease can be fatal within a few hours. Infections: Scope of
Toxic shock syndrome associated with the use of menstrual
tampons can be prevented by the appropriate use of tampons, the Problem
including washing hands thoroughly before and after inserting
Learning Outcomes
a tampon, using tampons with the lowest practical absorbency,
changing tampons about every 6 hours and using a pad instead of 5. Discuss behaviors that increase the risk of acquiring STIs.
a tampon while sleeping. It is also important to avoid trauma to the 6. List three ways that STIs can be prevented.
vagina when inserting tampons, to recognize the signs and symp-
toms of staphylococcal toxic shock syndrome, and to remove any
tampon immediately if symptoms occur. Women who have had MicroByte
Despite spending billions each year for STI control, an estimated 19
staphylococcal toxic shock syndrome previously should not use million Americans are infected annually, almost half in individuals
tampons. Table 25.5 describes the main features of staphylococ- 15 to 24 years old.
cal toxic shock syndrome.
 Part IV Infectious Diseases 621

FIGURE 25.7 The Possible Risk of Acquiring STIs

in People Having Unprotected Sex Each partner
had two previous sexual partners, and each of these
partners had two previous partners, and so on. This
risk of contracting an STI rises with the number of
sexual partners. (HSV-2—herpes simplex virus type 2,
HIV—human immunodeficiency virus, HPV—human
papillomavirus)
Each of these agents
can give asymptomatic ? What measures can be taken to avoid getting an STI?
infections:
HSV-2
Neisseria gonorrhoeae
HIV acquiring an STI, although does not absolutely guarantee
HPV
Chlamydia trachomatis
protection.
Treponema pallidum The list of diseases that can be transmitted sexu-
ally is very long and includes some that have non-sexual
modes of transmission as well—for example, shigellosis,
giardiasis, scabies, and viral hepatitis. The epidemiology can
be obscure, such as when a drug abuser contracts hepatitis B
from sharing a contaminated needle, remains symptom-free
for years, and then unknowingly transmits the virus to another
person during sexual intercourse. Table 25.7 lists some common
STIs discussed in this chapter. shigellosis, p. 588 giardiasis, p. 602
hepatitis, pp. 599–601
A person who gets one STI may have acquired others without
knowing it, so he or she needs to be tested for that possibility.
The risk of acquiring an STI increases sharply with the number of
partners with whom an individual has unprotected sex, and with Signs and Symptoms
TABLE 25.6 That Suggest an STI
the numbers of sexual partners of those partners (figure 25.7).
Table 25.6 lists some often-overlooked signs and symptoms 1. Abnormal discharge from the vagina or penis
that can indicate the possibility of an STI. These symptoms
2. Pain or burning sensation with urination
require clinical evaluation even if they go away without treatment,
especially if they appear within a few weeks of sexual intercourse 3. Sore or blister (painful or painless) on the genitals or nearby;
swellings in the groin
or other intimate sexual contact with a new partner. A number of
STIs can cause few or no signs or symptoms, and yet they can 4. Abnormal vaginal bleeding or unusually severe menstrual cramps
have serious effects and are transmissible to other people. 5. Itching in the vaginal or rectal area
Simple measures are highly effective in preventing STIs. 6. Pain in the lower abdomen in women; pain during sexual
These include abstaining from sexual intercourse or having a intercourse
monogamous relationship with a non-infected person. Using latex 7. Skin rash or mouth lesions
or polyurethane condoms also significantly reduces the risk of

TABLE 25.7 Common Sexually Transmitted Infections

Disease Cause Comment

Bacterial
Gonorrhea Neisseria gonorrhoeae Average reported cases per year—340,000. True incidence much higher.
Chlamydial infections Chlamydia trachomatis Average reported cases per year—800,000. True incidence much higher.
Syphilis Treponema pallidum Average reported cases (primary and secondary) per year—6,600.
Chancroid Haemophilus ducreyi Average reported cases per year—50. True incidence much higher.
Viral
Genital herpes Herpes simplex virus (HSV) Not reportable. Estimated 45 million Americans infected; about 85%
HSV, type 2.
Papillomavirus infections Human papillomavirus (HPV) Not reportable. Estimated 40 million Americans infected.
AIDS Human immunodeficiency virus (HIV) Average reported cases per year—40,000.
Protozoal
Trichomoniasis (“trich”) Trichomonas vaginalis Not reportable. Estimated 5 million Americans infected per year.
622 Chapter 25 Genitourinary Tract Infections

MicroAssessment 25.4
An individual who gets an STI may also have unknowingly acquired
others as well. The chance of a person acquiring an STI increases
significantly with the number of his or her sexual partners, and with
the number of sexual partners of those partners. Signs and symptoms
of an STI can sometimes easily go unnoticed or be ignored. Simple
measures are highly effective for preventing STIs.
10. Name two diseases that have both sexual and non-sexual modes
of transmission.
11. Discuss two ways in which STIs can be avoided.
12. Why should an individual with an STI be checked for other
STIs even if he or she has no other signs or symptoms? +

25.5 ■ Bacterial STIs

Learning Outcomes
7. Compare and contrast gonorrhea and chlamydial genital system
infections.
8. Compare and contrast syphilis and chancroid.

Most of the bacteria that cause STIs survive poorly in the environ-
ment. Because of this, transmission from one person to another
usually requires intimate physical contact and is highly unlikely
to occur by a handshake or from a toilet seat. Most STI pathogens FIGURE 25.8 Urethral Discharge in a Man with Gonorrhea
are adept at avoiding both innate and adaptive immunity. While symptoms of gonorrhea in men are noticeable when present,
the disease is often asymptomatic.

Gonorrhea ? What is the risk of asymptomatic infection?

Gonorrhea is the second most commonly reported STI in the

United States. The name originates from the Greek words gonos tissue blocks the tubes that carry the sperm, or if testicular tissue
meaning “seed” and rhoia meaning “to flow,” probably because is destroyed by the infection, infertility can result.
the pus produced during infection looks like semen. If gonorrhea is untreated in women, the infection can progress
Gonorrhea is not a new problem and was originally believed upward through the uterus into the fallopian tubes, causing pelvic
to be a symptom of syphilis, another common STI. In the mid– inflammatory disease (PID) and sometimes infertility. Scarring of
nineteenth century, gonorrhea was recognized as a specific disease. a fallopian tube can also lead to ectopic pregnancy, in which the
Today, it is still common, and its increasing resistance to antibacte- embryo develops in the fallopian tube or even in the abdominal
rial treatment is a cause for concern. cavity outside the uterus. Ectopic pregnancy can lead to life-
threatening internal hemorrhaging. Occasionally, the infection
Signs and Symptoms spreads from the fallopian tubes into the abdominal cavity, where
The incubation period of gonorrhea is usually only 2 to 5 days. it can affect the liver or other abdominal organs. It is unclear
Infections are often asymptomatic in both men and women. If geni- how N. gonorrhoeae (which is non-motile) can move through the
tal signs and symptoms occur in men, they include urethritis, pain uterus to reach the fallopian tubes, but it is possible that the organ-
during urination, and a thick, pus-containing discharge from the isms are carried on sperm, to which the bacteria attach.
penis (figure 25.8). These symptoms are noticeable and unpleas- Occasionally, the causative agent of gonorrhea can produce
ant, and men who have them usually go immediately for treatment. disseminated gonococcal infection (DGI). DGIs are character-
Gonorrhea follows a different course in women. Genital signs and ized by fever, rash, and arthritis caused by growth of the pathogen
symptoms if present include painful urination and vaginal dis- within the joint spaces. They can also cause endocarditis and men-
charge. The causative agent grows well in the cervix and fallopian ingitis. DGIs are not usually preceded by urogenital symptoms
tubes, in glands in the vaginal wall, and in other areas of the genital and are caused by strains of the organism that are serum-resistant
tract, causing inflammation, pain, and possible scarring. (see Causative Agent). endocarditis, p. 672 meningitis, p. 643
Untreated gonorrhea in either men or women can lead to Ophthalmia neonatorum is a destructive N. gonorrhoeae
complications. In men, an inflammatory reaction to the infection infection of the eyes of newborn babies whose mothers have
can cause scar tissue formation that partially obstructs the urethra, symptomatic or asymptomatic gonorrhea. The bacteria are trans-
slowing urination and predisposing the man to UTIs. The infection mitted during the infant’s passage through the infected birth canal.
may spread to the prostate gland and testes, producing prostatic Signs and symptoms include irritation of the conjunctiva and
abscesses and orchitis (inflammation of the testicles). If scar production of thick pus.
 Part IV Infectious Diseases 623

Pathogenesis
Neisseria gonorrhoeae has many mechanisms that allow it to
avoid host defenses. The organism is a human-specific pathogen
that survives poorly in the environment but is well adapted for col-
onization and proliferation in the host. Infection begins when the
organism selectively attaches by means of pili (fimbriae) and Opa
(outer membrane) proteins to receptors on certain non-ciliated
epithelial cells of the body, including those of the urethra, uterine
cervix, mouth, pharynx, anus, and conjunctiva. GC cannot colo-
nize squamous epithelium, such as that lining the adult vagina,
or ciliated cells. The bacteria enter the epithelial cells by endocy-
tosis, mediated by a porin protein in the LOS membrane. LOS
and peptidoglycan released by autolysis of cells cause comple-
ment activation, triggering an inflammatory response that leads to
most of the symptoms of gonorrhea. They also stimulate the pro-
duction of pro-inflammatory cytokines such as tumor necrosis fac-
Neutrophil Neisseria 15 μm tor (TNF) that cause the release of enzymes such as proteases and
gonorrhoeae phospholipases, leading to cell and tissue damage. porin, p. 60
FIGURE 25.9 Neisseria gonorrhoeae Stained smear of pus from GC is very good at avoiding both innate and adaptive immune
the urethra showing N. gonorrhoeae. responses. For example, it (1) produces proteases that destroy IgA
? What is the morphology of N. gonorrhoeae? found on mucosal surfaces; (2) uses host sialic acid to make a
capsulelike structure that allows the organism to resist phagocyto-
sis; and (3) produces an outer membrane porin protein that allows
Causative Agent it to survive within phagocytes by preventing phagolysosome
Gonorrhea is caused by the gonococcus (GC) Neisseria formation. Furthermore, LOS is also antigenically similar to red
gonorrhoeae—a fastidious Gram-negative diplococcus that blood cells, and this similarity to “self” cells prevents an effective
requires a rich medium such as chocolate agar for cultivation immune response.
(figure 25.9). Some strains of the organism produce pili (fimbriae) N. gonorrhoeae avoids adaptive immunity by antigenic and
for attachment to host cells (figure 25.10). The outer membrane phase variation. It can express several antigenic types of LOS and
of GC has the general structure of a typical Gram-negative LPS Opa proteins (antigenic variation), or it may not produce any at
membrane. However, the lipid that makes up the membrane has all (phase variation). The organism can change the type of LOS
a highly branched oligosaccharide structure, and is referred to as they express by an unknown mechanism. A single strain of GC
lipooligosaccharide (LOS). LOS plays a role in GC virulence and can also express many different kinds of pili, brought about by
pathogenicity. N. gonorrhoeae is very efficient at obtaining iron chromosomal rearrangements within the pili genes. Furthermore,
(needed for bacterial invasion) from the host during growth. It does the organisms may produce pili or not. Finally, GC Opa proteins
this by producing several membrane proteins that take iron from allow the organism to attach specifically to receptors on T helper
transferrin and lactoferrin under low iron conditions. cells, preventing their activation and proliferation. antigenic
variation, p. 178 phase variation, p. 178

Epidemiology
Gonorrhea is among the most common of the STIs. In the United
States, its incidence is the highest of any reportable bacterial
disease other than Chlamydia infection. The number of cases has
been declining since the emergence of HIV/AIDS, however, prob-
ably due to increased condom use.
N. gonorrhoeae infects humans only, living mainly on the
mucous membranes of the host. Most strains are susceptible to UV
light, cold and desiccation and so do not survive well outside the
host. For this reason, gonorrhea is transmitted almost exclusively
by direct contact. Because the bacteria mainly live in the genital
tract, this contact is almost always sexual. However, gonorrhea
can be transmitted by vaginal, oral, or anal sex. It can also be
passed from mother to baby during vaginal childbirth. Factors that
Pili 0.5 μm influence the incidence of gonorrhea include:
FIGURE 25.10 Pili on Single Coccus of Neisseria ■ Birth control pills. Oral contraceptives without use of a
gonorrhoeae (EM) condom offer no protection against STIs and may increase
? What is the function of the pili? susceptibility to them. Use of oral contraceptives leads to
624 Chapter 25 Genitourinary Tract Infections

migration of gonorrhea-susceptible epithelial cells from the and fluoroquinolones. A single class of antibiotics, the cephalo-
cervical lumen onto more exposed areas of the outer cervix. sporins, is currently recommended for treatment of gonorrhea.
Oral contraceptives also tend to increase both the pH and the Ciprofloxacin is commonly used.
moisture content of the vagina, favoring infection with GC Prevention of gonorrhea depends on abstinence, monogamous
and other agents of STIs. Besides being more susceptible to relationships, and consistent, correct use of condoms. Rapid iden-
gonorrhea, women taking oral contraceptives are also more tification and treatment of sexual contacts also prevents gonorrhea
likely to develop serious complications from the disease. by decreasing the overall number of infected individuals. There is
■ Asymptomatic infection. Male and females who have no vaccine for preventing gonorrhea because the antigenic varia-
asymptomatic gonorrhea can unknowingly transmit the infec- tion of the causative organism and the lack of an animal model for
tion over months or even years. Up to 20% of men and 60% the disease have made it impossible to develop one.
of women with gonorrhea are asymptomatic. Ophthalmia neonatorum is prevented by putting an antibiotic
ointment such as erythromycin directly into the eyes of all new-
■ Lack of immunity. There is little or no immunity following
born infants within 1 hour of birth. Some mothers who feel certain
recovery from the disease. Individuals can contract gonorrhea
that they do not have gonorrhea have challenged this practice, but
repeatedly.
because gonorrhea is frequently asymptomatic and the disease can
cause blindness in infants, this prophylactic treatment of a baby’s
Treatment and Prevention eyes is required by law. As a result, ophthalmia neonatorum is
N. gonorrhoeae is now frequently resistant to many antibacterial now unusual in the United States. Table 25.8 describes the main
medications, including penicillins, tetracyclines, sulfonamides, features of the disease.

TABLE 25.8 Gonorrhea

1 Neisseria gonorrhoeae enters Signs and Men: pain on urination,

the body through mucous symptoms discharge; complications include
membranes of the genitalia, impaired urinary flow, sterility,
mouth, or anus. It can also 1 or arthritis. Women: pain on
enter the eyes; serious urination, discharge, fever,
infections leading to loss of pelvic pain; sterility, ectopic
vision are likely in newborns. pregnancy, arthritis can occur.
2 The cervix is the usual site of Men and women may be
primary infection in women. asymptomatic.
6 Incubation period 2 to 5 days
3 The outer covering of the liver
is infected when gonococci Causative agent Neisseria gonorrhoeae (GC), a
enter the abdominal cavity 3 Gram-negative diplococcus
from infected fallopian tubes.
Pathogenesis Organisms attach to certain
4 Prostatic gonococcal abscesses non-ciliated epithelial cells
may be difficult to eliminate. by pili; phase and antigenic
5 Infection of the fallopian tubes 5 variation in surface proteins
results in scarring, which can 2 allows attachment to
cause infertility or ectopic 1 different host cells and escape
pregnancy. 7 from immune mechanisms.
Inflammation, scarring; can
6 Organisms carried by the spread by bloodstream.
bloodstream infect the heart
valves and joints. Epidemiology Transmitted by sexual contact.
7 4 Asymptomatic carriers. No
7 Urethral scarring from 2 immunity.
gonococcal infection can
predispose to urinary infections 8 Treatment Treatment: ceftriaxone.
by other organisms. and prevention Prevention: abstinence,
monogamous relationships,
8 Scarring of testicular tubules condoms, early treatment of
can cause sterility. sexual contacts.
 Part IV Infectious Diseases 625

Chlamydial Infections cell-mediated immune response. The cell wall of C. trachomatis

lacks peptidoglycan but instead has cysteine-rich proteins. This
Chlamydial infections are among the most common STIs world-
wall inhibits the formation of a phagolysosome, allowing the
wide. These infections resemble gonorrhea because they can cause
organism to survive within phagocytes. endocytosis, p. 72
urethritis and may lead to testicle and fallopian tube damage that
Chlamydial infection usually involves the urethra in both
result in infertility. Worldwide, they are the leading cause of infer-
men and women. In men, it spreads to the epididymis, causing
tility. There are several different antigenic types of the causative
acute pain and swelling. It may lead to infertility. In women, the
agent that lead to distinct diseases. Chlamydia, p. 277
infection commonly involves the cervix, making it bleed easily,
especially with sexual intercourse. The uterus also often becomes
Signs and Symptoms infected, leading to pain and bleeding. From the uterus, the infec-
The signs and symptoms of Chlamydia trachomatis infection gen- tion is carried by sperm to the fallopian tubes, where it can lead
erally appear 7 to 14 days after exposure. In men, the main symp- to PID (pelvic inflammatory disease), causing the risk of ectopic
tom is a thin, gray-white discharge from the penis, sometimes pregnancy or infertility (figure  25.11). Like N. gonorrhoeae,
with painful testes. Some men also develop pain on urination, and C. trachomatis can move from the fallopian tubes to the abdomi-
fever. Women with genital chlamydia most commonly develop an nal cavity and infect the surface of the liver.
increased vaginal discharge, sometimes accompanied by painful
urination, abnormal vaginal bleeding, upper or lower abdominal Epidemiology
pain, and pain during sexual intercourse. Many infections of men Chlamydial genital infections are the most common of all notifiable
and women are asymptomatic—nearly 75% of women and 50% bacterial infectious diseases. The number of reported cases has been
of men with chlamydia do not develop signs or symptoms and rising, probably because of increased awareness of the disease and
can have the disease for months or years before being diagnosed. better diagnostic tests. Non-sexual transmission of this agent also
Some strains of C. trachomatis can cause lymphogranuloma occurs, for example, in non-chlorinated swimming pools. Newborn
venereum, a rare disease in which the lymph nodes of the groin babies of infected mothers often develop chlamydial ophthalmia
swell and drain pus. Enlarged nodes, called buboes, are commonly neonatorum and pneumonia from C. trachomatis infection con-
painful. Repeated draining and healing of buboes eventually leads tracted during passage through the birth canal.
to fibrosis, which can obstruct lymphatic vessels, resulting in
gross swelling of the genitalia after several years. The long-term Treatment and Prevention
effect of this symptom may be genital elephantiasis.
Several antibiotics can be used to treat chlamydial infections.
MicroByte Azithromycin can be given as a single dose, whereas tetracyclines
A type of chlamydial infection of the eyes, called trachoma, can and erythromycin require more doses but are less expensive alter-
cause blindness if left untreated. natives. Sexual partners are also treated. Chlamydial ophthalmia
neonatorum is treated with oral erythromycin.
Chlamydial infections can be prevented by abstinence,
Causative Agent monogamous relationships, and condoms used correctly. All sexu-
Chlamydia trachomatis is a spherical, obligate intracellular Gram- ally active people are advised to get tested for Chlamydia each
negative bacterium. During replication, the organism causes
inclusion bodies containing a glycogen-like material to form in
the cytoplasm of host cells. These inclusions stain with iodine and
can provide a fast way of identifying C. trachomatis infections.
Microvilli
Different antigenic types of C. trachomatis cause distinct
diseases. Approximately eight types are responsible for most
C. trachomatis STIs. Three other types cause lymphogranuloma
venereum and four others cause trachoma, a serious eye infection. C. trachomatis
glycogen, p. 32

Pathogenesis
The infectious form of C. trachomatis, called an elementary
body, attaches specifically to receptors on the surface of the
host epithelial cell, inducing the cell to take in the bacterium by
endocytosis. Once inside the endosome, the elementary body
interacts with glycogen inclusions, enlarges, and becomes a non-
infectious reticulate body. The reticulate body divides repeat- 2 μm

edly by binary fission, resulting in many new elementary bodies. FIGURE 25.11 Chlamydia trachomatis Scanning electron
These are released when the host cell lyses, and infect nearby micrograph showing C. trachomatis attached to fallopian tube
cells. The infected cells produce cytokines that cause an intense mucosa.
inflammatory reaction. Much of the tissue damage results from the ? Is Chlamydia limited to infections of the genital mucosa?
626 Chapter 25 Genitourinary Tract Infections

Chlamydial Genital FIGURE 25.12

TABLE 25.9 Syphilitic Chancre
System Infections A chancre develops
at the site where
Signs and Men: thin, gray-white penile discharge,
Treponema pallidum
symptoms painful testes. Women: vaginal discharge,
entered the body, as
vaginal bleeding, lower or upper
on the foreskin of this
abdominal pain
uncircumcised penis.
Incubation period Usually 7 to 14 days
? What is the risk
Causative agent Chlamydia trachomatis, an obligate associated with
intracellular bacterium, certain serotypes an open ulcer, as
Pathogenesis Elementary body attaches to specific shown here?
receptors on the epithelial cell, causing
endocytosis; becomes reticulate body in the
endosome; repeated replication by binary
fission and differentiation into elementary
bodies; host cell bursts, releasing
elementary bodies to infect adjacent cells;
release of cytokines results in inflammatory
response; cell-mediated immune response
against infection causes extensive damage;
scar tissue forms, causing ectopic pregnancy
and infertility.
■ Primary syphilis. The stage is characterized by a painless,
Epidemiology The leading reportable bacterial infection red ulcer called a hard chancre (pronounced “shanker”) that
in the United States. Large numbers of
asymptomatic men and women carriers.
appears at the site of infection about 3 weeks after exposure
Non-sexual transmission can occur in non- (figure  25.12). The chancre usually develops on the genitalia
chlorinated swimming pools. but may occur anywhere on the body. The local lymph nodes
Treatment Treatment: azithromycin and other enlarge. Primary syphilis chancres often go unnoticed in women
and prevention antibacterial medications. Prevention: and men who have sex with men because they are hidden from
abstinence, monogamous relationship, view in the vagina or anus and are painless. Sometimes no chan-
condom use. Test sexually active people at cre develops, only a pimple small enough to go unnoticed.
least once yearly to rule out asymptomatic
infection. ■ Secondary syphilis. The symptoms of secondary syphilis
usually appear 2 to 10 weeks (or longer) after the primary
stage. The infection has spread systemically by this time, and
causes many diverse signs and symptoms, the most common
year, or twice yearly if they have multiple partners or if their part- of which is a rash that includes the palms and soles, and white
ner has multiple partners. The main features of chlamydial genital patches on the mucous membranes (figure 25.13). Other signs
infections are summarized in table 25.9. and symptoms include runny nose and watery eyes, aches and
pains, sore throat, fever, malaise, weight loss, and headache.
Syphilis Mucosal patch
During the first half of the twentieth century, syphilis was a major
cause of mental illness and blindness, and a significant cause of heart
disease and stroke. The disease was almost eradicated by the mid-
1950s through a successful program aimed at locating and treating
all people with syphilis and their sexual contacts. Unfortunately, the
number of cases then increased in the 1990s due to factors including
inner-city poverty, prostitution, and drug use. This trend was reversed
through renewed efforts in education, case finding, and treatment
that caused a dramatic drop in the number of new cases. The AIDS
epidemic added urgency to syphilis control efforts, because syphilis,
like other STIs that cause genital sores, promotes the spread of AIDS
by increasing the risk of HIV infection. By the end of 1998, the syphilis
rate was at the lowest level since it became a notifiable disease. Rates
have risen since then, particularly in women (see Perspective 25.1).

Signs and Symptoms FIGURE 25.13 Secondary Syphilis Secondary syphilis lesions
Syphilis causes so many different signs and symptoms that it is such as these oral mucous patches contain numerous Treponema
easily confused with other diseases and is often called “the great pallidum organisms and are highly infectious.
imitator.” Its symptoms occur in defined clinical stages. ? What causes the manifestations of secondary syphilis?
 Part IV Infectious Diseases 627

Occasionally, hepatitis (liver disease) and renal disease occur.

The person is most infectious at this stage.
■ Latent syphilis. During this stage, the person does not have
any signs and symptoms of syphilis, although they have
antibodies to the causative agent. A quarter of people in this
stage recover; another quarter remain in this stage. The rest
progress to the next stage of the disease.
■ Tertiary syphilis. After a latent period that can last for many
years, signs and symptoms of tertiary syphilis sometimes
occur. The symptoms of this stage can be grouped into three
categories. In gummatous syphilis, localized areas of tis-
sue damage develop as a result of prolonged inflammatory
responses. These lesions, called gummas, are granulomas
similar to the tubercles seen in tuberculosis. Gummas are
chronic and may occur anywhere in the body (figure 25.14). FIGURE 25.15 Hutchinson Teeth Notice the notched, deformed
incisors, a late manifestation of congenital syphilis.
Cardiovascular syphilis is characterized by aneurysms that
form in the ascending aorta, caused by chronic inflammation ? What are some other manifestations of congenital syphilis?
of the arterioles supplying this vessel. Neurosyphilis occurs
in a small number of cases and is characterized by a pat- include runny nose, rash, and liver and spleen enlargement. Late
tern of symptoms including personality change, emotional symptoms occur after 2 years. The bones, cartilage, and teeth can
instability, delusions, hallucinations, memory loss, impaired become deformed as the child develops, causing cleft palate, sabre
judgment, abnormalities of the pupils of the eye, and speech shins (bent tibias), saddle nose (sunken nasal bridge), mulberry
defects. Other signs and symptoms of this stage include blind- molars (many cusps), and Hutchinson teeth (notched incisors)
ness, vertigo, insomnia, stroke, and meningitis. Although (figure 25.15). Eye inflammation and deafness often occur.
neurosyphilis symptoms are usually seen during the tertiary
stage, they can also be experienced during the first and sec- MicroByte
ondary stages of the disease. tubercles, p. 504 Evidence suggests that T. pallidum is directly involved in aiding
HIV infection and progression. Stopping syphilis would decrease the
During pregnancy, T. pallidum easily crosses the placenta and number of new HIV cases.
infects the fetus, causing congenital syphilis. Fetal infections
can occur in the absence of any signs or symptoms of syphilis
in the mother and at any stage of pregnancy, but damage to the Causative Agent
fetus does not generally happen until the fourth month. Common Syphilis is caused by Treponema pallidum, a very slender, motile
outcomes of fetal infection include spontaneous abortion, still- spirochete that is difficult to see unless dark-field microscopy is
birth, and neonatal death. Almost all infected infants that survive used (figure  25.16). All spirochetes are highly motile, and their
birth develop characteristic signs and symptoms. Early signs and endoflagella—which propel them in a corkscrewlike motion—
symptoms resemble those of severe adult secondary syphilis and allow them to penetrate a wide variety of tissues and organs. The

5 μm
FIGURE 25.14 A Gumma of Tertiary Syphilis Gummas are
formed by an inflammatory mass which can perforate tissue, as in FIGURE 25.16 Treponema pallidum This organism must be
this example in the nose. viewed with dark-field microscopy.
? Where in the body do gummas occur? ? Why is dark-field microscopy needed to observe T. pallidum?
628 Chapter 25 Genitourinary Tract Infections

PERSPECTIVE 25.1
The Death of Syphilis?
For 40 years (1932 to 1972), the U.S. Public Forty wives became infected, and there were Syphilis is a good target for eradication
Health Service conducted a study (now known 19 children born with congenital syphilis. because it has no animal reservoir, infectious
as the Tuskegee Syphilis Experiment) of Nothing of scientific value was learned, and cases can generally be treated with a single
399 poor Alabama black men with advanced many people to this day remain bitter and injection of penicillin, and the incubation
syphilis. The study was done to assess the distrustful of the U.S. Public Health Service. period is long enough that sexual contacts of
natural progression of the disease in black In 1997, President Clinton, speaking for the an infected person can be found and treated
men. The men were not told their diagnosis government, formally apologized to the sur- before they spread the disease any further.
nor educated about the nature of their condi- viving eight subjects. Moreover, instead of being widely endemic,
tion and its transmission. For the first half Fortunately, in more recent years, major most syphilis cases are now concentrated in a
of the study, there were no effective drugs governmental efforts to conquer the disease relatively few areas and certain populations.
against syphilis, and so the men remained have met with some success. In 2009, only Also, new techniques are now available for
untreated. However, even after effective ther- 13,997 cases of primary and secondary syphilis rapid diagnosis, treatment, and epidemiologi-
apy became available, the study was allowed were reported, down from 50,223 in 1990. The cal tracing of the disease. Finally, control of
to continue without giving treatment, in a bla- Centers for Disease Control and Prevention syphilis is closely linked to AIDS control
tant mix of bad science and racism. Twenty- (CDC) has even considered the possibility that efforts because the risk of HIV transmission
eight of the men died of the disease, and transmission of the disease could be eliminated is six times as great if either sex partner has
another hundred died from its complications. in the United States. syphilis.

organism is fragile and easily killed by heat, cold, desiccation, year, and then gradually subsides. About 50% of untreated cases
and soap. It is therefore transmitted almost exclusively by sex- never progress past the secondary stage. After a latent period of
ual or oral contact. In nature, T. pallidum infects only humans. from 5 to 20 years or even longer, however, some people with the
dark-field microscopy, p. 43 disease develop tertiary syphilis. immune complexes, p. 395
T. pallidum must be grown in the testicles of laboratory rab- Tertiary syphilis signs and symptoms occur from hypersensi-
bits to study its cell division and pathogenesis, because it does not tivity reactions to small numbers of T. pallidum that grow and per-
replicate in vitro. The organism lacks metabolic abilities, and does sist in the tissues. In this stage, the patient is no longer infectious.
not have enzymes needed for the TCA cycle or electron transport The organisms may be present in almost any part of the body,
chain, so cannot generate much ATP. It is therefore thought to get and the signs and symptoms of tertiary syphilis depend on where
most of its essential macromolecules from the host, instead of syn- the hypersensitivity reactions occur. If they occur in the skin,
thesizing them itself. The sequence of nucleotides in its genome bones, or other areas not vital to existence, the disease is not life-
is now known, which should give scientists a better understanding threatening. If, however, they occur within the walls of a major
of its virulence, why it cannot be cultivated in vitro, and how to blood vessel such as the aorta, the vessel may become weakened
make an effective vaccine. TCA cycle, p. 142 and even rupture, resulting in death. The main characteristics of
the stages of syphilis are summarized in table 25.10.
Pathogenesis
Treponema pallidum easily penetrates mucous membranes and dam- Epidemiology
aged skin. The infectious dose is very low—less than 100 organisms. Syphilis is usually transmitted by sexual intercourse. However,
In primary syphilis, T. pallidum multiplies in a localized area of the infection can occur from kissing a person with secondary syphilis,
genitalia, spreading from there to the lymph nodes and bloodstream.
The hard chancre is caused by an intense inflammatory response to
the bacteria, which are present in high numbers in the lesion. The TABLE 25.10 Stages of Syphilis
chancre disappears within 2 to 6 weeks, even without treatment, and
Stage of
patients may mistakenly believe that they have recovered from the Disease Main Characteristics Infectious?
disease. However, the organism avoids destruction by the body’s
defenses and progression of the disease can continue for years. Few Primary Firm, painless ulcer (hard Yes
virulence factors have been discovered for T. pallidum, and the outer chancre) at site of infection;
lymph node enlargement
membrane of the cells lack LPS, the molecule that normally triggers
an immune response. There are also few proteins in the outer mem- Secondary Rash, aches, and pains; rash Yes
and mucous membrane
brane and therefore few targets for opsonizing antibodies to bind.
lesions
Many of the signs and symptoms of secondary syphilis are
Latent No signs or symptoms; Early—yes
due to immune complexes that form as specific antibodies bind to
serologically positive Late—no
circulating T. pallidum. By this time, the spirochetes have become
Tertiary Gummas; damage to large No
systemic, and infectious lesions occur on the skin and mucous
blood vessels, eyes, nervous
membranes in various locations, especially in the mouth. The system; insanity
secondary stage lasts for weeks to months, sometimes as long as a
 Part IV Infectious Diseases 629

or by contact with a primary ulcer infected with T. pallidum.

Because there is no animal reservoir, eradicating this disease is
possible. To do this, however, cases must be identified and treated,
particularly in high-risk groups such as prostitutes, men who
have sex with men, and jail inmates. Infected individuals can be
detected using a simple blood test.

Treatment and Prevention

Primary and secondary syphilis are easily treated with an antibiotic
such as penicillin—the dose and drug type depend on the stage of
the disease. Treatment is more difficult for tertiary syphilis, because
most of the organisms are not actively multiplying and penicillin
acts against dividing cells. Congenital syphilis can be prevented
by diagnosing and treating the mother’s syphilis before the fourth
month of pregnancy, when T. pallidum starts to affect the fetus.
There is no vaccine for syphilis. Abstinence, monogamous
relationships, effective and correct use of condoms, and other
(a) (b)
safer sex practices decrease the risk of contracting the disease.
Quick identification and treatment of sexual contacts are impor- FIGURE 25.17 Chancroid (a) Lesions of the penis. These
tant in limiting spread of the disease. Table 25.11 summarizes the ulcerations are soft and painful. (b) Swollen groin lymph nodes.
The nodes are tender, often pus-filled, and may break open
main features of syphilis. and drain.

Chancroid ? How are chancroid ulcers different from chancres?

Chancroid is another bacterial STI that, like syphilis, causes geni-

tal sores, thereby increasing the risk of contracting HIV. The dis-
ease is common in developing countries, usually associated with Signs and Symptoms
commercial sex workers. The reported incidence of chancroid in Chancroid is characterized by one or more painful genital sores
the United States is low, but it often goes unreported or misdiag- called soft chancres (figure  25.17a). Typically, these begin as a
nosed, and may therefore be more common than recorded. small pimple at the site of bacterial entry through the skin, but

TABLE 25.11 Syphilis

1 Treponema pallidum enters the Signs and Chancre, fever, rash, stroke,
body through a microscopic 6 symptoms nervous system deterioration;
abrasion or mucous membranes, can imitate many other diseases
usually genitalia, mouth, or
1 Incubation period 10 to 90 days
rectum.
2 4 Causative agent Treponema pallidum, a non-
2 A chancre develops at site of
culturable spirochete
entry.
3 Organisms multiply locally and Pathogenesis Primary lesion, or chancre,
appears at site of inoculation,
spread throughout the body by
heals after 2 to 6 weeks;
the bloodstream. 6
T. pallidum invades the blood
4 Infectious mucous patches and 3 vessel system and is carried
skin rashes of secondary syphilis throughout the body, causing
appear. A fetus may become fever, rash, mucous membrane
infected, resulting in miscarriage lesions; damage to brain,
or a live-born infant with 4 arteries, and peripheral nerves
congenital syphilis. appears years later.
5 An asymptomatic latent period Epidemiology Sexual contact with infected
occurs. T. pallidum disappears partner; kissing; transplacental
from blood, skin, and mucous passage
4
membranes. 1 Treatment Treatment: penicillin. Prevention:
6 After months or years, 2 and prevention abstinence, monogamous
symptoms of tertiary syphilis relationships, use of condoms,
appear: heart and blood vessel safer sex practices, treatment of
defects, gummas, strokes, sexual contacts, reporting cases.
eye abnormalities, general
neurological symptoms, insanity.
630 Chapter 25 Genitourinary Tract Infections

they then ulcerate and quickly get bigger. The lymph nodes in the MicroAssessment 25.5
groin also enlarge and become tender (figure 25.17b). Sometimes
the nodes are pus-filled and may rupture, discharging the pus. In Transmission of bacterial STIs usually requires direct person-to-
person contact. Unsuspected STIs (such as gonorrhea and syphilis)
women, chancres typically occur on the labia or thighs, causing of pregnant women are a serious risk to fetuses and newborn babies.
pain during urination and sexual intercourse. Even asymptomatic infections can cause genital tract damage
(including PID) and can be spread to other people.
Causative Agent 13. Can a baby have congenital syphilis without its mother ever
Chancroid is caused by Haemophilus ducreyi, a fastidious, pleo- having had signs and symptoms of syphilis? Explain.
morphic, Gram-negative coccobacillus that, like N. gonorrhoeae, 14. What, if any, is the relationship between chancroid and
can only be cultivated on a rich medium such as chocolate agar. development of AIDS?
Haemophilus, p. 276 15. Why does scarring of a fallopian tube increase the risk
of an ectopic pregnancy? +
Pathogenesis
Haemophilus ducreyi infection causes an intense inflammatory
response, with accumulation of neutrophils and macrophages in the 25.6 ■ Viral STIs
area. The bacteria survive by producing proteins that prevent phago-
cytes from engulfing them. In addition, proteins in the outer mem- Learning Outcomes
brane protect the cells from activated complement proteins. The 9. Compare the infections caused by genital herpes simplex virus
mechanism of these protective proteins is still being investigated. and human papillomavirus.
10. Outline the relationship between HIV infection and AIDS.
Epidemiology
Epidemics in American cities associated with prostitution some- Viral STIs are probably as common or more common than the
times occur. In some tropical countries, chancroid is second only bacterial diseases, and they are incurable. Their effects can be
to gonorrhea in prevalence of the STIs. severe and long-lasting. Genital herpes can cause recurrent signs
and symptoms for years, papillomavirus infections can lead to
Treatment and Prevention cancer, and untreated HIV infections usually result in AIDS.
Chancroid can usually be treated with a variety of antibiotic
options, including erythromycin, azithromycin, or ceftriaxone.
Genital Herpes
Some strains, however, are resistant to multiple antibacterial
medications. Effectiveness of therapy is also significantly reduced Genital herpes is among the top three or four most common STIs.
if the patient has AIDS. An estimated 45 million Americans are infected with the causative
Chancroid can be prevented by abstinence from sexual virus, and about 500,000 new cases occur each year.
intercourse, monogamous relationships, correct use of condoms,
and safer sex practices. Table  25.12 gives the main features of Signs and Symptoms
chancroid. Genital herpes begins 2 to 20 days (usually about a week) after
exposure, with genital itching and burning, and in some cases,
severe pain. Groups of small, red bumps appear on the genitalia
or anus, depending on the site of infection. These bumps become
TABLE 25.12 Chancroid blisters surrounded by redness (figure 25.18). The blisters break
Signs and One or more painful, gradually enlarging,
in 3 to 5 days, leaving an ulcerated area that slowly dries and
symptoms soft chancres on or near the genitalia; large, becomes crusted. This eventually heals without a scar. Less com-
tender regional lymph nodes mon signs and symptoms include discharge from the penis or
Incubation period 3 to 10 days
Causative agent Haemophilus ducreyi, a pleomorphic,
fastidious Gram-negative rod requiring special
growth media FIGURE 25.18
Genital Herpes
Pathogenesis A small pimple appears first, which ulcerates on the Shaft of
and gradually enlarges; multiple lesions may the Penis
join together; lymph nodes enlarge, liquefy,
and may discharge to the skin surface. ? Can genital
herpes be
Epidemiology Sexual transmission. Common in prostitutes;
transmitted
fosters the spread of AIDS.
when there are
Treatment Treatment: several antibacterial medications no visible signs
and prevention effective. Resistance can be a problem. or symptoms of
Prevention: abstinence, monogamous the infection?
relationships; avoiding sexually promiscuous
partners; correct use of condoms.
 Part IV Infectious Diseases 631

vagina, headache, fever, muscle pain, and in the case of women, because herpes lesions can occur on parts of the genitalia that a
painful urination. Signs and symptoms are the worst in the first 1 condom does not cover.
to 2 weeks and disappear within 3 weeks. Genital herpes signs and Genital herpes can be a serious risk to newborn babies. If
symptoms often recur, however, because the virus becomes latent. the mother has a primary infection near the time of delivery, the
The recurrent signs and symptoms are usually not as severe as baby has about a 30% risk of acquiring the infection. The baby
those of the first episode, and recurrences usually occur less often often dies from the infection or is permanently disabled by it. To
with time. Some individuals have recurrences throughout their prevent contact with the infected birth canal, these babies must be
life, whereas others never have recurrence. latent infections, p. 383 delivered by cesarean section. If the mother has recurrent disease,
the risk to the baby is very low, presumably because transplacental
Causative Agents anti-HSV antibody from the mother protects the baby, but doctors
Genital herpes is usually caused by herpes simplex virus type 2 will often do a cesarean section to minimize the risk. The main
(HSV-2), an enveloped, double-stranded DNA virus. Herpes sim- features of genital herpes are presented in table 25.13.
plex virus type 1, the cause of “cold sores” (“fever blisters”) can
also cause genital herpes. The two virus types look the same, and
their genomes are about 50% homologous. Either virus can infect
Papillomavirus STIs: Genital Warts
the mouth and the genitalia, but HSV-2 causes more severe genital and Cervical Cancer
lesions with a greater frequency of recurrence. HSV-1, p. 582 Sexually transmitted human papillomavirus (HPV) strains
are among the most common of the STI agents, infecting
Pathogenesis an estimated 40 million Americans. Some HPV strains cause
Lesions begin with infection of a group of epithelial cells that papillomas—warty growths of the external and internal genitalia.
lyse following viral replication, creating small, fluid-filled blisters
(vesicles) containing large numbers of infectious virions. The
vesicles burst, producing painful ulcers. During initial infection TABLE 25.13 Genital Herpes
and for as long as 1 month after, the virus is found in genital secre-
Signs and Itching, burning pain at the site of
tions. In recurrence, the virus is usually present in large numbers symptoms infection, painful urination, tiny blisters
for less than a week. with underlying redness. The blisters break,
Latency of the disease is not completely understood but prob- leaving a painful superficial ulcer, which
ably depends on cell-mediated immunity as well as viral products. heals without scarring. Recurrences are
common.
Most of the time, the viral DNA exists within nerve cells in a
circular, non-infectious form, causing no symptoms. In this state, Incubation Usually 1 week (range, 2 to 20 days)
period
only a single gene is expressed, and it encodes for a small segment
of RNA called a microRNA (miRNA), that probably causes Causative agent Usually herpes simplex virus type 2. The
latency of the virus. At times, however, the entire viral chromo- cold sore virus, herpes simplex type 1, can
also be responsible. Herpesviruses are
some can be transcribed and complete infectious virions produced. enveloped and contain double-stranded
These reinfect the area supplied by the nerve and cause symptom DNA.
recurrence.
Pathogenesis Lysis of infected epithelial cells results in
fluid-filled blisters containing infectious
Epidemiology virions. Vesicles burst, causing a painful
HSV-2 can survive for short periods on fomites or in water, but ulcer. The acute infection is controlled by
immune defenses; genome persists within
non-sexual transmission is rare. Sexual transmission of HSV is
nerve cells in a non-infectious form beyond
most likely to occur during the first few days of symptomatic dis- the reach of immune defenses. Replication
ease, but it can happen in the absence of signs or symptoms of the of infectious virions can occur and cause
disease. Once infected, an individual is forever at risk of transmit- recurrent symptoms in the area supplied
ting the virus to another person. HSV, like other ulcerating genital by the nerve. Newborn babies can contract
fatal generalized herpes infection if their
diseases, promotes the spread of AIDS by increasing the risk of
mother has a primary infection at the time
HIV infection. Neither HSV-1 nor HSV-2 has animal reservoirs. of delivery.
Epidemiology No animal reservoirs. Transmission by
Treatment and Prevention sexual intercourse, oral-genital contact.
There is no cure for genital herpes, although anti-HSV medica- Transmission risk greatest first few days of
tions such as acyclovir and famciclovir can decrease the severity active disease. Transmission can occur in
of the first attack and the incidence of recurrences. the absence of symptoms. Herpes simplex
increases the risk of contracting HIV.
Genital herpes can be prevented by abstaining from sexual
intercourse or having a monogamous relationship with a genital Treatment and No cure. Medications help prevent
prevention recurrences, shorten duration of symptoms.
herpes-free person. In addition, avoiding sexual intercourse dur-
Prevention: abstinence, monogamy, and
ing active signs or symptoms and using condoms with a spermi- correct use of condoms help prevent
cide that inactivates HSVs help prevent contracting the disease. transmission.
However, using a condom does not always give full protection
632 Chapter 25 Genitourinary Tract Infections

Other strains cause non-warty lesions of mucosal surfaces such Causative Agents
as the uterine cervix, and these are a major factor in the develop- Human papillomaviruses are naked, double-stranded DNA viruses
ment of cervical cancer. of the papovavirus family. There are more than 100 types of
HPVs, and the different types are tissue- and species-specific. For
MicroByte
Between 75% and 80% of sexually active Americans will have an
example, HPVs that cause the common warts of the hands and
HPV infection at some point during their life. feet generally do not infect the genitalia. Similarly, the HPVs that
cause genital warts may infect the cervix but do not cause cancer.
At least 40 HPV types are transmitted by sexual contact.
Signs and Symptoms Approximately 15 of these are strongly associated with cancer of
Most people who have HPV infections clear them without ever the cervix, penis, anus, vagina and throat. These high-risk HPVs
developing signs or symptoms. In fact, they often do not know include HPVs 16, 18, 31, and 45.
that they are infected. Some types of HPV do cause symptoms,
however, and warts are the most easily recognized of these. Pathogenesis
Warts usually appear about 3 months after infection (range,
HPVs are thought to enter and infect the deeper layers of epi-
3 weeks to 8 months), developing on the head or shaft of the penis
thelium through microscopic abrasions. The genome of low-risk
(figure  25.19), at the vaginal opening, or around the anus.
(wart-causing) HPV types exists in infected cells as extrachromo-
Different kinds of lesions develop depending on the virus type
somal, closed DNA circles, but the mechanism by which warts
and the location of the infection. Lesions can be flat or raised,
arise is unknown. In contrast, the genome of high-risk (cancer-
cauliflowerlike, or hidden within the epithelium. Occasionally,
associated) HPV types can integrate into the chromosome of the
warts become inflamed and bleed. After warts are removed, HPV
host cell; they are oncogenic because they code for a protein that
persists in surrounding normal-looking epithelium and can cause
allows excessive cell growth. Oncogenic types of HPV tend to
additional warts. Warts sometimes partly block the urethra or, if
cause infections that persist longer than other HPV types, and
very large, the birth canal. Newborn infants can become infected
to cause precancerous lesions. It takes on average 20 years for
with HPV at birth and develop warts that block their respiratory
the infection to cause invasive cancer. A cancer-associated HPV
tract, a serious condition that occurs occasionally (less than one
is present in almost 100% of cervical cancers, but only a small
out of every 100,000 births). Certain HPV strains can cause cervi-
percentage of infections by these viruses result in cancer. This
cal cancer. Precancerous lesions are generally asymptomatic and
indicates that other unknown factors must be present for cancer to
can be detected only by examining the cervical tissues. The same
develop. oncogenes, p. 323
strains can also cause other cancers including oral, penile, vaginal,
and anal cancers. Accumulating evidence indicates that oral sex is
a risk factor for oral cancers. A CDC study implicated HPV as the Epidemiology
cause of up to 60% of mouth and throat cancers. HPV strains are easily spread by sexual intercourse. Asymptomatic
individuals infected with HPV can transfer the virus to others.
HPV infection is the most common reason for an abnormal
Papanicolaou (Pap) test in teenage women (see next section). A
history of having multiple sex partners is the most important risk
FIGURE 25.19
Genital Warts factor for acquiring genital HPV infection.
Penis with warts,
a manifestation
of human
Treatment and Prevention
papillomavirus Warts often regress over time, but several treatments are available
infection. to remove them or hasten their clearance. Unfortunately, none of
? Would these methods cure the infection, so the warts may recur. Dermal
condom warts can be removed by laser treatment, freezing with liquid
use always nitrogen, or surgical excision. External genital warts can be treated
prevent the with creams and ointments, including (1) imiquimod (Aldara
transmission
of HPV?
Cream), an immune modifier that induces cytokine production;
(2) podofilox, which prevents cell division of infected cells; and
(3) sinecatechin-containing ointments, which cause wart regres-
sion by an unknown mechanism.
HPV infections can be prevented by abstinence, having
monogamous relationships, or using condoms appropriately. As
with HSV, condoms do not provide complete protection against
HPV because the virus can be transmitted by exposure to areas
not covered by the condom. Women should have a Pap smear
every 12 months. This test involves removing cells from the cer-
vix, staining them on a microscope slide, and examining them for
 Part IV Infectious Diseases 633

Abnormal cells
TABLE 25.14 Papillomavirus STIs
Signs and Often asymptomatic. Warts of the external
symptoms and internal genitalia the most common
symptom. Symptoms are strain-specific.

Incubation Usually 3 months (range, 3 weeks to

period 8 months)
Causative agents Human papillomaviruses, many types,
naked DNA viruses of the papovavirus
family. Different types infect different
tissues and produce different lesions.

Pathogenesis Virus enters epithelium through abrasions,

infects deep layer of epithelium; establishes
latency; cycles of replication occur when
host cell begins maturation; cancer-
associated viral types can integrate into
the host cell chromosome and can cause
FIGURE 25.20 Abnormal Papanicolaou (Pap) Smear The pink precancerous lesions.
and blue objects are squamous epithelial cells; abnormalities include
doubling of the nuclei and a clear area around them. Epidemiology Asymptomatic individuals can transmit the
disease; multiple sex partners the greatest
? What is the most frequent cause of an abnormal Pap smear in risk factor; warts can be transmitted to
young women? the mouth with oral sex, and to newborn
babies at birth.
Treatment Treatment: wart removal by multiple
and prevention techniques, but does not cure the infection.
any abnormal cells (figure  25.20). Cervical cancer is generally Prevention: latex condoms advised to
minimize transmission and avoiding sexual
preceded by precancerous lesions—areas of abnormal cell growth contact with people having multiple sex
that can be detected by this test. The lesion can be removed, partners. Pap tests at least yearly for sexually
thereby preventing development of a cancer. active women. Vaccines protect against
Two vaccines—Gardasil and Cervarix—are currently avail- some cancer-causing strains.
able that protect against infection by HPV types 16 and 18, which
are responsible for about 70% of cancers. Gardasil also protects
against HSV types 6 and 11, which cause more than 90% of
Signs and Symptoms
genital warts. Either vaccine is given in a three-dose series, and is
recommended for girls and young women ages 11 to 26. Although Six days to 6 weeks after contracting HIV, some individuals
not included in the recommended vaccine schedule, Gardasil is develop flulike signs and symptoms—fever, head and muscle
available for girls as young as 9, and for boys and men ages 9 aches, and enlarged lymph nodes. These signs and symptoms are
through 26. Cervarix is given to females only. Table 25.14 sum- often mild, and go away by themselves. Many individuals have no
marizes some important features of papillomavirus STIs. signs or symptoms at all. HIV infection then persists unnoticed for
almost 10 years before immunodeficiency develops, with certain
malignancies and unusual microbial infections that often attack
HIV/AIDS the lungs, intestines, skin, eyes, or the central nervous system.
Acquired immunodeficiency syndrome (AIDS) is unquestionably
the most important sexually transmitted infection of the past cen- Causative Agents
tury. It is covered briefly here and more fully in chapter 28. The Most AIDS cases are caused by human immunodeficiency virus
AIDS epidemic was first recognized in the United States in 1981. type 1 (HIV-1), an enveloped, single-stranded RNA virus of the
Over the first 15 years, 500,000 Americans developed AIDS, and retrovirus family. Each virion contains the enzyme reverse tran-
300,000 died of the disease. With major advances in prevention scriptase and two copies of the viral genome. In parts of western
and treatment of the disease, and expenditures of huge amounts Africa, another retrovirus, HIV-2, is a common cause of AIDS; it
of money, the epidemic shows signs of leveling off. Nevertheless, is rarely a cause in the United States. retroviruses, p. 320
in 2009, an estimated 33.3 million people around the world were
infected with human immunodeficiency virus (HIV), the caus- Pathogenesis
ative agent of AIDS, and more than 2 million died of AIDS. An HIV can attack a variety of cell types in the human body, but most
estimated 1.7 million new infections were in sub-Saharan Africa critically affects the helper T (TH) cells. These cells have the CD4
alone. The advances in prevention and treatment of AIDS are not surface protein to which the virus attaches, and they are referred to
uniformly available to many of those around the world who suffer as CD4 + cells. In order for the virus to enter the cell, it must also
because of the disease. AIDS, p. 695 attach to certain cytokine receptors, such as CCR5 or CXCR4 on
634 Chapter 25 Genitourinary Tract Infections

the host cell surface. After the virus enters the CD4+ cell, it makes are also successful. Supplying injected-drug abusers with sterile
a DNA copy of its RNA genome using its reverse transcriptase. needles and syringes in exchange for used ones decreases trans-
The DNA copy then integrates into the cell’s genome using the missions of the virus without encouraging addiction. Educational
virally encoded enzyme integrase. Once integrated, the virus is programs aimed at minimizing high-risk sexual practices have also
inaccessible to antiviral chemotherapy. shown short-term success in decreasing HIV spread. In addition,
If the CD4+ cell becomes activated, the viral genes are tran- identifying and treating other STIs and promoting consistent use
scribed, creating RNA copies. These serve dual roles, function- of latex condoms decreases the spread of HIV.
ing both as mRNA molecules that can be translated and RNA The World Health Organization has reported that circumci-
genomes for new viral particles. The new viral particles bud sion (surgical removal of the penis foreskin) can reduce HIV
out of the host cell, ultimately leading to the death of that cell. incidence by up to 60% and should also be included in prevention
Despite the body’s ability to replace hundreds of billions of CD4+ strategies. Circumcision changes the anatomy of the penis, for
lymphocytes, the number of these lymphocytes slowly declines example, reducing the number of Langerhans cells, which are HIV
over many months or years. Infection of macrophages, which are targets. It also changes the normal microbiota populations under
also CD4+ cells, adds to the decline in cell-mediated immunity. the foreskin, reducing the number of certain anaerobic species,
Although macrophages are generally not killed by HIV, their some of which increase the risk of HIV infection by enhancing
function is impaired both by the viral infection and the lack of virus stability. Another promising prevention strategy is pre-
interaction with TH lymphocytes. Eventually, the immune system exposure prophylaxis, in which ARVs are given before sexual
becomes so impaired it can no longer respond to infections or intercourse in an effort to reduce transmission (see chapter 28).
cancers. helper T cells, p. 356 macrophages, p. 340 About 25% of the estimated 900,000 people in the United
States infected with HIV are unaware of their infection and more
Epidemiology than half of new HIV infections in the country are caused by those
HIV is present in blood, semen, and vaginal secretions in unaware of their HIV status. Individuals who, since the 1980s,
symptomatic and asymptomatic infections. The virus spreads have engaged in injected drug use or risky sexual behavior, and
horizontally mainly by sexual intercourse and sharing hypodermic anyone having had unprotected sexual intercourse with them,
needles. It is not highly contagious, and most transmissions of this should get a blood or saliva test to rule out HIV infection. People
kind can be prevented by changes in human behavior. HIV can who learn that their HIV test is positive can receive optimum treat-
also be transmitted vertically—from mother to newborn—during ment sooner and also prevent transmission of the virus to others.
pregnancy, during passage through the birth canal, and through Table 25.15 presents the main features of HIV disease and AIDS.
breast-feeding. horizontal and vertical transmission, p. 440
TABLE 25.15 HIV Disease and AIDS
Treatment and Prevention
Signs and No symptoms, or flulike symptoms early in
Treatment of HIV disease is designed to block replication of the symptoms the illness; an asymptomatic period typically
virus, but it does not affect viral nucleic acid already integrated in lasting years; symptoms of lung, intestine, skin,
the genome of host cells. Four groups of antiretroviral medications eyes, brain, and other infections, and certain
are currently available—those that (1) block reverse transcriptase cancers
activity, (2) act against viral protease, (3) block viral integrase, Incubation About 6 days to 6 weeks for flulike symptoms;
and (4) interfere with viral attachment to host cells. The first three period many months or years for cancers and unusual
infections
block enzymes essential at different stages of viral replication.
The fourth type is represented by a drug that interferes with HIV Causative agents Generally human immunodeficiency virus type
1 (HIV-1)
attachment to the host cell cytokine receptor CCR5. Viral mutants
resistant to a single medication, however, quickly arise. To help Pathogenesis The virus infects CD4+ lymphocytes and
macrophages, thereby slowly destroying the
prevent the selection of resistant variants, a “cocktail” of several ability of the immune system to fight infections
medications, each acting in a unique way, is given. When combi- and cancers.
nations of antiviral drugs are given, the treatment is referred to as Epidemiology HIV present in blood, semen, and vaginal
highly active antiretroviral therapy (HAART). In many cases, secretions in symptomatic and asymptomatic
this therapy can clear the patient’s blood of detectable viral nucleic infections; spread usually by sexual intercourse,
acid, stop the progress of the disease, and even allow partial recov- sharing of needles by injected-drug abusers,
ery of immune function. Therapy is expensive and the medications and from mother to infant at childbirth. Other
STIs foster transmission.
often have serious side effects, two features that preclude use of the
regimen in many of the world’s HIV disease sufferers. Although Treatment Treatment: reverse transcriptase and protease
and prevention inhibitors in combination (HAART). Prevention:
not curative, chemotherapy can significantly prolong and improve abstinence from sexual intercourse and drug
the quality of life. HIV replication, p. 699 HAART, p. 703 abuse; monogamy; consistent use of latex
Prevention of HIV disease is aimed at education and safer sex condoms; avoidance of sexual contact with
practices. One of the most important developments in AIDS pre- injected-drug abusers, those with multiple
vention was the finding that vertical transmission from mother to partners or history of STIs. Anti-HIV medication
for pregnant women and their newborn infants.
newborn can be stopped in about two-thirds of the cases by using Circumcision and pre-exposure prophylaxis.
antiretroviral medications (ARVs). Needle-exchange programs
 Part IV Infectious Diseases 635

MicroAssessment 25.6 some cases, there is burning pain with urination. The vulva and
vaginal wall are red and slightly swollen, and may develop pin-
Viral STIs are as common as bacterial STIs, but so far they are point hemorrhages. Symptomatic men have penile discharge, burn-
incurable. Genital herpes is common and, like most other STIs,
can be transmitted in the absence of signs or symptoms. Human
ing pain with urination, painful testes, or a tender prostate gland.
papillomaviruses cause genital warts; some play an important role in
cancer of the cervix and probably cancers of the vagina, penis, anus, Causative Agent
and throat. HIV disease generally ends in AIDS, but treatment can Trichomoniasis is caused by Trichomonas vaginalis, a motile
significantly improve longevity and quality of life. protozoan that has four anterior flagella and a posterior flagellum
16. What are the possible consequences of sexually transmitted attached to an undulating membrane (figure 25.21). It also has a
papillomavirus infections for men, women, and babies? slender, posteriorly protruding, rigid rod called an axostyle that is
17. What kinds of sex partners present a high risk of transmitting used for attachment. T. vaginalis can be identified by its unmistak-
HIV? able, jerky motility on microscopic examination of the vaginal
18. Why should a person be concerned about genital herpes—is drainage. Unlike most other pathogenic protozoa, T. vaginalis
it not just a cold sore on the genitals? Explain. + lacks a cyst form to aid its survival in the environment away from
the host’s body. The organism is a eukaryote, but it does not have
any mitochondria. Instead, it has interesting cytoplasmic organelles
called hydrogenosomes for glucose metabolism and respiration.
25.7 ■ Protozoal STIs Enzymes within these double membrane-bound organelles remove
Learning Outcome the carboxyl group (COOH) from pyruvate and transfer electrons
to hydrogen ions, producing hydrogen gas. mitochondria, p. 74
11. List the distinctive characteristics of the organism that causes
trichomoniasis.
Pathogenesis
A number of protozoan diseases can be transmitted sexually. The pathogenesis of trichomoniasis is not fully understood.
Most, however, are intestinal infections that can be transmitted T. vaginalis produces a variety of adherence factors (including
through oral-anal contact during sexual activity. Trichomoniasis is adhesins, microtubules, and microfilaments) specific for vaginal
a sexually transmitted protozoan disease that involves the genital epithelium, allowing it to colonize the vulva and vagina walls.
system. protozoa, p. 291 The signs and symptoms of trichomoniasis are thought to be due
to mechanical trauma by the axostyle of the moving protozoa,
but toxins or enzymes such as cysteine proteinases might also be
Trichomoniasis (“Trich”) involved. The frothy discharge is probably due to the gas pro-
Trichomoniasis ranks third after bacterial vaginosis and vulvo- duced by the organisms.
vaginal candidiasis among the diseases that commonly cause vagi-
nal signs and symptoms. Men can also be infected. An estimated Epidemiology
7.4 million Americans contract this STI each year. Trichomonas vaginalis is distributed worldwide as a human
parasite and has no other reservoirs. It is easily killed by drying,
Signs and Symptoms so transmission is usually by sexual contact. The organism can
Trichomoniasis is often asymptomatic, especially in men. survive for a time on moist objects such as towels and bathtubs,
Symptomatic infections in women are characterized by itching of however, so it can occasionally be transmitted non-sexually.
the vulva and inner thighs, itching and burning of the vagina, and Nevertheless, T. vaginalis infections in children should at least
a frothy, sometimes smelly, yellowish-green vaginal discharge. In raise the question of sexual abuse and possible exposure to other

Anterior flagella FIGURE 25.21 Trichomonas

vaginalis T. vaginalis is a common
cause of vaginitis.
? What is unusual about the
Undulating metabolism of this organism?
membrane
Nucleus

Axostyle

5 μm
636 Chapter 25 Genitourinary Tract Infections

STIs. Newborn infants can contract the infection from infected TABLE 25.16 Trichomoniasis
mothers at birth. The high percentage of asymptomatic infections,
Signs and Women: itching, burning, swelling, vaginal
especially in men, helps transmission of the disease. Infection
symptoms redness; frothy, sometimes malodorous,
rates are highest in men and women with multiple sex partners. yellow-green discharge, and burning on
Infection with T. vaginalis causes tissue damage, so people with urination. Men: discharge from penis,
this disease are at higher risk of contracting HIV. burning on urination, painful testes,
tender prostate. Many women, most men
MicroByte asymptomatic
An African study indicated that trichomoniasis can cause a two- to
Incubation 4 to 20 days
threefold increase in HIV transmission. period
Causative agent Trichomonas vaginalis, a protozoan with
Treatment and Prevention four anterior flagella, and a posterior
flagellum attached to an undulating
Most strains of T. vaginalis respond quickly to metronidazole membrane; a rigid rodlike structure
treatment, but a few are resistant. Transmission is prevented by called an axostyle protrudes posteriorly;
abstinence, monogamy, and the use of condoms. Table  25.16 unmistakable jerky motility; no
gives the main features of trichomoniasis. mitochondria; hydrogenosomes
are present
The key features of the diseases covered in this chapter are high- Pathogenesis Unexplained. Inflammatory changes and
lighted in the Diseases in Review 25.1 table. pinpoint hemorrhages suggest mechanical
trauma from the motile organisms.
MicroAssessment 25.7 Epidemiology Worldwide distribution; asymptomatic
carriers foster spread; easily killed by drying
Trichomoniasis is a protozoan STI that involves the genital tract. The
due to lack of cyst form, transmission by
causative agent is a flagellated protozoan that does not form cysts intimate contact; high rate of infection
and lacks mitochondria. Organelles called hydrogenosomes produce with multiple sex partners. Newborn
hydrogen gas from pyruvate. infants of infected mothers can acquire the
19. What significance does the lack of a cyst form of Trichomonas infection at birth.
vaginalis have on the epidemiology of trichomoniasis? Treatment Treatment: metronidazole. Prevention:
20. What is the relationship between being “easily killed by drying” and prevention abstinence, monogamy, and consistent use
and “transmission usually by sexual contact”? + of condoms.

FUTURE CHALLENGES 25.1

Getting Control of Sexually Transmitted Infections
Few problems are as complicated as getting to therapeutic agents helps in choosing prompt perhaps using new polymers or antimicrobial
control of STIs because of the psychological, treatment. vaginal gels.
cultural, religious, and economic factors that Using condoms is an effective method Effective vaccines could block the spread
are involved—factors that vary from one pop- for interruption of STI transmission, but in of STIs, but their development has been
ulation and culture to another. Gaining control practice there are a number of problems with extremely problematic and remains a long-
means focusing on diagnosis, interruption of using them. When infected college students term goal. Sequencing the genomes of the
transmission, education, and treatment. The are asked why they did not use a condom, they causative agents is helping to identify appro-
challenge is to develop innovative approaches often state that “it takes away the romance,” “it priate antigens to include in vaccines, but
in each of these areas and to apply them world- decreases pleasurable sensation,” or “that’s for using them to stimulate a protective immune
wide on a sustained basis. sissies.” Also, many individuals are allergic response is a continuing challenge.
In the area of diagnosis, the challenge to latex, and the alternatives, except for poly- Educational efforts focusing on groups at
is to use molecular biological techniques to urethane, are unreliable for preventing disease high risk for STIs have had mixed success.
identify and produce specific antigens and transmission. Worldwide, as many as one out Education can be a useful tool for gaining
antibodies that can be used to quickly identify of three condoms is defective, and people in control of STIs, and the challenge is to better
causative agents and determine the extent of the poorest countries cannot even afford them. understand the reasons for its failures.
their spread. The development of nucleic acid The challenge is to develop cheaper, more
probes to identify genes coding for resistance reliable, and more acceptable barrier methods,
Diseases in Review 25.1
Genitourinary Infections

Disease Causative Agent Comment Summary Table

UROGENITAL INFECTIONS

Cystitis Usually uropathogenic Most common in women because of their relatively short urethra; Table 25.1, p. 615
(bladder Escherichia coli characterized by painful urination; can progress to kidney
infection) infection.
Leptospirosis Leptospira interrogans Spread by water contaminated with the urine of infected animals; Table 25.2, p. 617
enters via mucous membranes or breaks in skin, and then spreads
to all tissues, including eyes.
Bacterial Unknown, possibly Characterized by thin, gray-white discharge and fishy odor. Most Table 25.3, p. 618
vaginosis (BV) bacterial common in sexually active women and pregnant women.

Staphylococcal Toxin-producing strains Toxin is a superantigen and causes cytokine release, leading to Table 25.5, p. 620
toxic shock of Staphylococcus drop in blood pressure; an epidemic occurred as a result of
aureus high-absorbency tampon use.

BACTERIAL STIS

Gonorrhea Neisseria gonorrhoeae Infections are often asymptomatic, particularly in women. Signs Table 25.8, p. 624
and symptoms include painful urination and pus (men) or vaginal
discharge; can progress to PID.
Chlamydial Chlamydia trachomatis Most common STI in the United States; infections are often Table 25.9, p. 626
infections asymptomatic; can progress to PID.
Syphilis Treponema pallidum Primary syphilis is characterized by a painless chancre, secondary Table 25.10, p. 628
by a widespread rash, and tertiary by cardiovascular and nervous Table 25.11, p. 629
system damage. Congenital infections also occur.
Chancroid Haemophilus ducreyi Characterized by painful genital sores and enlarged groin lymph nodes. Table 25.12, p. 630

VIRAL STIS

Genital herpes Herpes simplex virus, Characterized by itching, burning, painful blisters. Virus becomes Table 25.13, p. 631
usually type 2 (HSV-2) latent, so infections are lifelong.
Papillomavirus Human papillomaviruses Some strains cause genital warts, and others cause cancers. Warts Table 25.14, p. 633
infections (HPVs) removed using lasers, liquid nitrogen, or immune modifier creams.
Vaccines protect against some cancer-causing strains.
HIV/ AIDS Human immunodeficiency HIV disease usually progresses to AIDS. Virus infects helper T cells, Table 25.15, p. 634
virus, usually type 1 (HIV-1) resulting in immunodeficiency. HAART slows disease progression.

FUNGAL DISEASE

Vulvovaginal Candida albicans Characterized by intense itching and thick, white discharge. Table 25.4, p. 619
candidiasis Associated with antibiotics or other conditions that disrupt the
(VVC) normal vaginal microbiota, allowing the overgrowth of C. albicans.

PROTOZOAL STI

Trichomoniasis Trichomonas vaginalis Sexually transmitted; characterized by itching, discharge, and Table 25.16, p. 636
(“trich”) urinary discomfort.
638 Chapter 25 Genitourinary Tract Infections

Summary
25.1 ■ Anatomy, Physiology, and Ecology Staphylococcal Toxic Shock Syndrome (table 25.5)

The Urinary System (figure 25.1) Staphylococcal toxic shock syndrome became widely known with
a 1980 epidemic in menstruating women who used a certain kind
The urinary system is composed of the kidneys, ureters, bladder, and
of vaginal tampon that has since been removed from the market
urethra. Frequent urination with complete bladder emptying is an
(figure  25.6). Signs and symptoms include sudden fever, headache,
important defense mechanism against UTI. Infections occur more fre-
muscle aches, bloodshot eyes, vomiting, diarrhea, a sunburnlike rash
quently in women than in men because of the shortness of the female
that later peels, and confusion. The blood pressure drops, and without
urethra and its closeness to the opening of the intestinal tract.
treatment, kidney failure and death may occur.
The Genital System (figure 25.2)
The fallopian tubes, which are open on both ends, provide a passageway 25.4 ■ Sexually Transmitted Infections:
for infection to enter the abdominal cavity. The uterine cervix is a fre- Scope of the Problem (tables 25.6, 25.7)
quent site of infection and a place where cancer can develop. The vagina In the United States, 19 million new STIs occur each year, almost half
is a portal of entry for a number of infections. In men, the prostate can in individuals 15 to 24 years old. Simple measures exist for controlling
enlarge and partially obstruct urinary flow. STIs: abstinence from sexual intercourse, a monogamous relationship
The lower urethra is inhabited by various microorganisms, some- with an uninfected person, and consistent and correct use of latex or
times including potential pathogens. The normal vaginal microbiota is polyurethane condoms (figure 25.7).
affected by estrogen hormones, which cause deposition of glycogen in
the cells lining the vagina. Vaginal lactobacilli metabolize the glycogen 25.5 ■ Bacterial STIs
and release lactic acid and hydrogen peroxide, thereby protecting the Most of the bacteria that cause STIs survive poorly in the environment
vagina from colonization by pathogens. and require intimate contact for transmission.
Gonorrhea (table 25.8)
25.2 ■ Urinary Tract Infections Gonorrhea, caused by Neisseria gonorrhoeae (figures  25.9, 25.10), is
Any condition that prevents a person from urinating normally increases one of the most commonly reported bacterial diseases. Men usually
the risk of infection. The urinary system usually becomes infected by develop painful urination and thick pus draining from the urethra
organisms ascending from the urethra, but it can also be infected from (figure  25.8). Women may have similar signs and symptoms, but these
the bloodstream. tend to be milder and are often unnoticed. Inflammatory reaction to
the infection causes scarring, which can partially obstruct the urethra
Bacterial Cystitis (“Bladder Infection”) (table 25.1)
or cause infertility in men and women. Expression of different surface
Most UTIs in healthy people are caused by Escherichia coli or antigens allows attachment of N. gonorrhoeae to different types of cells.
other Enterobacteriaceae members from the person’s own normal Antigenic variation has prevented development of a vaccine.
intestinal microbiota. Healthcare-associated UTIs are common and
are caused by Pseudomonas aeruginosa, Serratia marcescens, and Chlamydial Infections (table 25.9, figure 25.11)
Enterococcus faecalis, which are often resistant to many antibiotics. Chlamydial genital infections, caused by Chlamydia trachomatis, are
Kidney infection (pyelonephritis) may complicate untreated bladder reported more often than any other bacterial disease. Signs and symp-
infection when pathogens ascend through the ureters to the kidneys. toms and complications of chlamydial infections are very similar to
those of gonorrhea, but are milder. Asymptomatic infections are com-
Leptospirosis (table 25.2)
mon and easily transmitted. Sexually active people should be tested at
In leptospirosis, the urinary system is infected by Leptospira interrogans least once a year so that transmission can be stopped and complications
from the bloodstream (figure  25.3). This biphasic illness causes fever, prevented.
bloodshot eyes, and pain in the septicemic phase. Improvement occurs,
then new signs and symptoms emerge, with damage to multiple organs Syphilis (tables 25.10, 25.11)
during the immune phase. Many species of animals are chronically Syphilis, caused by Treponema pallidum, (figure  25.16), is called “the
infected with L. interrogans and excrete the organism in their urine. great imitator” because its many signs and symptoms can resemble
other diseases. Primary syphilis is characterized by a painless, firm
25.3 ■ Genital System Diseases ulceration called a hard chancre; the organisms multiply and spread
throughout the body (figure  25.12). In secondary syphilis, skin and
Bacterial Vaginosis (BV) (table 25.3, figure 25.4) mucous membranes show lesions that have large numbers of the organ-
Bacterial vaginosis is the most common cause of vaginal signs and isms (figure 25.13); a latent period of months or years occurs between the
symptoms that include a gray-white discharge from the vagina and a secondary and tertiary phases of the disease. Tertiary syphilis is not con-
strong fishy odor; there is no inflammation. The causative agent or tagious. It causes damage to the eyes and the cardiovascular and central
agents are unknown, but BV occurs with a disruption of the normal nervous systems. An inflammatory, necrotizing mass called a gumma
microbiota. can involve any part of the body (figure  25.14). Syphilis in pregnant
women can spread across the placenta to involve the fetus, resulting in
Vulvovaginal Candidiasis (VVC) (table 25.4)
congenital syphilis (figure 25.15).
Vulvovaginal candidiasis is second among causes of vaginal disorders. Signs
and symptoms include itching, burning, vulvar redness and swelling, and a Chancroid (table 25.12)
thick, white discharge. The causative agent is a yeast, Candida albicans, Chancroid is another widespread bacterial STI, but it is not com-
that is commonly part of the normal vaginal microbiota (figure 25.5). Anti- monly reported because of difficulties in making a bacterial diagnosis.
bacterial treatment, uncontrolled diabetes, and oral contraceptives are Epidemics in the United States have been associated with prostitution.
predisposing factors, but in most cases no such factor can be identified. Chancroid is caused by Haemophilus ducreyi, and is characterized by
 Part IV Infectious Diseases 639

single or multiple soft, tender genital ulcers, and enlarged, painful groin (figure 25.20). Several
types of HPV, including HPV 16 and 18, are asso-
lymph nodes (figure  25.17). Some strains of H. ducreyi are resistant to ciated with cancer of the cervix, vagina, penis, anus, or throat.
various antimicrobial drugs, making treatment more difficult.
HIV/AIDS (table 25.15)
AIDS is the end stage of disease caused by human immunodeficiency
25.6 ■ Viral STIs virus (HIV). Worldwide, millions of new HIV infections and deaths
Viral STIs are at least as common as bacterial STIs, but they are not from AIDS occur each year. HIV disease is usually first manifested
yet curable. as a flulike illness that develops about 6 days to 6 weeks after an indi-
vidual contracts the virus. An asymptomatic interval follows that typi-
Genital Herpes (table 25.13) cally lasts almost 10 years, during which the immune system is slowly
Genital herpes is a very common disease, important because of the and progressively destroyed. Unusual cancers and infectious diseases
discomfort and emotional trauma it causes, its potential for causing indicate the start of AIDS. No vaccine or medical cure is yet available,
death in newborn infants, and the increased risk it poses of transmitting but HIV transmission can be slowed significantly by consistent use of
HIV. Signs and symptoms may include a group of vesicles (figure 25.18) condoms and use of sterile needles by injected-drug abusers. A sig-
with itching, burning, or pain; these break, leaving painful ulcers. nificant reduction in mother-to-newborn transmission can be achieved
Local lymph nodes enlarge. Many infections have few or no signs or with medication. Pre-exposure prophylaxis with antiretroviral medica-
symptoms; some have painful recurrences. The virus establishes a latent tion also shows promise in reducing infection rates.
infection in sensory nerves and cannot be cured. It can be transmitted in
the absence of signs or symptoms, but the risk is greatest when lesions 25.7 ■ Protozoal STIs
are present.
Trichomoniasis (“Trich”) (table 25.16)
Papillomavirus STIs: Genital Warts and Cervical Cancer Signs and symptoms include itching, burning, swelling, and redness of
(table 25.14) the vagina; with frothy, sometimes smelly, yellow-green discharge; and
Papillomavirus STIs are probably more common than any other kind burning on urination. Men may have discharge from the penis, burning
of STI. The causative agents, human papillomaviruses (HPVs), are on urination, sometimes accompanied by painful testes and a tender
small DNA viruses. Genital infection symptoms are warts on or near prostate gland. Many women and most men are asymptomatic. The
the genitalia (figure 25.19). Some HPV types cause precancerous lesions causative agent is Trichomonas vaginalis, a protozoan with four anterior
that are asymptomatic and can be detected only by medical examina- flagella, and a posterior flagellum attached to an undulating membrane
tion. HPVs are the main cause of abnormal Pap smears in young women (figure 25.21).

Review Questions
Short Answer 2. Choose the one correct statement about leptospirosis.
1. Name two substances released by lactobacilli
ctobacilli that help protect the a) Humans are the only reservoir.
vagina from potential pathogens. b) Most infections produce severe symptoms.
2. List four things that predispose to thee development of infection of c) Transmission is by the fecal-oral route.
the urinary bladder. d) It can lead to unnecessary abdominal surgery.
3. Name two genera of bacteria that infect the kidneys from the e) Effective vaccine is generally available for preventing human
bloodstream. disease.
4. What possible danger can be found in a spot on the ground where 3. Which one of the following statements about bacterial vaginosis
an animal has urinated 1 week earlier? r? is false?
5. What is a clue cell? a) It is the most common vaginal disease in women of
6. What is ophthalmia neonatorum? childbearing age.
7. List three diseases caused by differentt antigenic types of Chlamydia b) In pregnant women, it is associated with a sevenfold increased
trachomatis. risk of obstetrical complications.
8. Why is dark-field microscopy used to o view Treponema pallidum? c) Inflammation of the vagina is a constant feature of the
9. Give two ways in which the chancre of chancroid differs from the disease.
chancre of syphilis. d) The vaginal microbiota shows a significant decrease in
10. What is the relationship between AIDS DS and HIV disease? lactobacilli and a marked increase in anaerobic bacteria.
e) The cause is unknown.
Multiple Choice 4. Pick the one false statement about vulvovaginal candidiasis.
1. Which of the following about bacterialial cystitis is false? a) It often involves the external genitalia.
a) About one-third of all women will have it at some time during b) It is readily transmitted by sexual intercourse.
their life. c) It is caused by a yeast present among the normal vaginal
b) Catheterization of the bladder markedly
rkedly increases the risk of microbiota in about one-third of healthy women.
contracting the disease. d) It is associated with prolonged antibiotic use.
c) Individuals who have a bladder catheter
theter in place indefinitely e) It involves increased risk late in pregnancy.
risk bladder infections with multiple
ple species of intestinal
bacteria at the same time.
d) Bladder infections occur as often in men as they do in women.
e) Bladder infections can be asymptomatic.
omatic.
640 Chapter 25 Genitourinary Tract Infections

5. All of the following statements about staphylococcal toxic shock 10. All of the following are true of “trich” (trichomoniasis) except
are true except a) It can cause burning pain on urination and painful testes in men.
a) It can quickly lead to kidney failure. b) It occurs worldwide.
b) The causative organism usually does not enter the bloodstream. c) Asymptomatic carriers are rare.
c) It occurs only in vaginal tampon users. d) Transmission can be prevented by proper use of condoms.
d) Almost one-third of victims of the disease will suffer a e) Individuals with multiple sex partners are at high risk of
recurrence sometime after recovery. contracting the disease.
e) Person-to-person spread does not occur.
6. Which of the following statements about gonorrhea is false? Applications
a) The incubation period is only a few days. 1. Religious restrictions of a small North African community are
b) Disseminated gonococcal infection (DGI) is almost invariably preventing a World Health Organization project from reducing the
preceded by prominent urogenital symptoms. incidence of gonorrhea. The community will not permit the testing
c) DGI can result in arthritis of the knee. of females for the disease. They can be treated, however, if they
d) Phase variation helps the causative organism evade the immune show outward evidence of the disease. Only males are allowed
response. to participate fully in the project, with testing for the disease and
e) Pelvic inflammatory disease (PID) is common in untreated treatment. The village elders argue that eradicating the disease
women. from males would eventually remove it from the population. What
would be the impact of these restrictions on the success of the
7. Which one of these statements about chlamydial genital infections
project?
is false?
2. Former President Ronald Reagan once commented at a press
a) The incubation period is usually shorter than in gonorrhea.
conference that the best way to combat the spread of AIDS in the
b) Infected cells develop inclusion bodies. United States was to prohibit everyone from having sexual contact
c) Pelvic inflammatory disease (PID) can be complicated by for 5 years. What would be the success of such a program if it were
infection of the surface of the liver. possible to carry it out?
d) Tissue damage largely results from cell-mediated immunity.
e) Fallopian tube damage can occur in the absence of symptoms. Critical Thinking +
8. Which symptom is least likely to occur as a result of tertiary 1. The middle curve of figure 25.6 shows the occurrence of staphy-
syphilis? lococcal toxic shock syndrome in menstruating women from 1979
a) Gummas b) White patches on mucous membranes to 2010. What aspect of these data argues that high-absorbency
c) Emotional instability d) Stroke tampons were not the only cause of staphylococcal toxic shock
syndrome associated with menstruation?
e) Blindness
2. In early attempts to identify and isolate the cause of syphilis, vari-
9. During the first 15 years of the AIDS epidemic, approximately
ous bacteria in the discharge from syphilitic lesions in experimen-
how many Americans died of the disease?
tal animals were isolated in pure culture. None of them, however,
a) 10,000 b) 50,000 would cause the disease when used in attempts to infect healthy
c) 300,000 d) 5 million animals. Why was it considered a critical step to have the culti-
e) 50 million vated bacteria reproduce the disease in the healthy animals?
26 Nervous System Infections
KKE EYYT TE ERRMMS S
Arbovirus Arthropod-borne RNA
virus, carried by vectors such as
mosquitoes.
Blood-Brain Barrier Cells that
Meninges Membranes covering
the brain and spinal cord.
Meningitis Inflammation of the
meninges.
work together to restrict exchange Peripheral Nervous System
between the bloodstream and (PNS) Division of the nervous
the brain. system that carries information to
Central Nervous System (CNS) and from the CNS.
Brain and spinal cord. Transmissible Spongiform
Cerebrospinal Fluid (CSF) Fluid Encephalopathy (TSE) Chronic
produced in the brain that flows degenerative brain disease caused
within and around the CNS. by prions; characterized by a spongy
appearance to brain tissue.
Encephalitis Inflammation of the
brain.

finally closed and converted to a military-style academy for high school

dropouts in 1999.
TEM of West Nile virus, a virus introduced into the United States in 1999.
Because the word leprosy carries centuries of dark overtones, many
people prefer to use the term Hansen’s disease, a name that honors the
discoverer of the causative bacterium.
A Glimpse of History

N
 
Today it is hard to appreciate the fear and loathing once attached to ervous system infections are frightening. They threaten
leprosy. The Bible refers to several disfiguring skin diseases, including a person’s ability to move, feel, or even think. Consider
leprosy (lepros, meaning “scaly”), and people suffering from the diseases
poliomyelitis, which can result in a paralyzed limb or
are portrayed as filthy, outcast, or condemned by God for sin. Moses
the inability to breathe without mechanical assistance. Hansen’s
called lepers “unclean” and proclaimed they must live away from others.
In the Middle Ages, lepers attended their own symbolic burial before disease (leprosy) can result in loss of fingers or toes or deformity
being sent away. of the face. Infections of the brain or its covering membranes can
Gerhard Henrik Armauer Hansen (1841–1912) was a burly Nor- render a child deaf or intellectually disabled. Before the discovery
wegian physician with many interests, ranging from science to religion of antibiotics, bacterial infections of the nervous system were
to polar exploration. When he was 32 years old, Hansen went into often fatal. Fortunately, CNS infections are uncommon.
medical research, and was named assistant to Dr. Daniel C. Danielson,
a leading authority on leprosy. Danielson believed that leprosy was a
hereditary disease of the blood, and considered the idea that the disease
was contagious as a “peasant superstition.” Hansen, however, disproved
26.1 ■ Anatomy, Physiology,
Danielson’s hypothesis in careful studies conducted over a number of and Ecology
years. He found a unique bacterium associated with the disease in every
leprosy patient he studied. His 1873 report of the findings marked the first Learning Outcomes
time that a specific bacterium was linked to a disease—almost a decade 1. Describe how information flows through and between neurons.
before Koch’s proof of the cause of tuberculosis.
2. Differentiate between the central nervous system and the
In the United States, even during the first half of the twentieth cen-
peripheral nervous system.
tury, persons diagnosed with leprosy risked having their houses burned
to destroy the source of infection. Their names were changed to avoid 3. Explain how bone, cerebrospinal fluid, meninges, and the
embarrassing their family, and they were whisked to a leprosarium such blood-brain barrier protect the central nervous system.
as the one at Carville, Louisiana, surrounded by a 12-foot fence topped
with barbed wire. Sufferers were separated from spouses and children and Nerve cells work together, transmitting electrical impulses
denied the right to marry or vote. Those who attempted to escape were throughout the body like a highly sophisticated circuit board.
captured and brought back in handcuffs. The Carville leprosarium was Each nerve cell, or neuron, has different regions with distinct
641
642 Chapter 26 Nervous System Infections

functions (figure 26.1a). Branching projections called dendrites evaluating the resulting loss of function. Generalized inflamma-
receive information. They convey that information to the cell tion or infection of the brain is called encephalitis.
body, the command center that contains the cell nucleus. A long, The peripheral nervous system (PNS) is made up of nerves
thin extension called an axon then transmits the information from composed of bundles of axons that carry information to and from
the cell body to another cell. the CNS. Axons of motor neurons carry messages from the CNS
Most neurons communicate using neurotransmitters, chem- to different parts of the body and cause them to respond; axons
icals produced in the cell body and stored in vesicles at the end of sensory neurons transmit sensations like touch, heat, light, and
of the axon. When neurotransmitters are released, the molecules sound to the central nervous system. Most nerves are mixed, car-
diffuse across the short distance to a neighboring cell. That cell rying both sensory and motor information throughout the body.
has receptors for the neurotransmitter, allowing it to receive and The cell bodies of sensory neurons from the skin are located in
respond to the signal. The region where transmission of infor- clusters called ganglia (singular: ganglion) near the vertebral col-
mation takes place between the two cells is called a synapse. umn; the cell bodies of motor neurons are located within the CNS.
Although movement of neurotransmitter molecules is normally The central nervous system is much better protected than
away from the cell body to the synapse, some viruses and toxins the peripheral nervous system. Both the brain and spinal cord are
can move through the axon toward the cell body in a process enclosed by bone—the brain by the skull and the spinal cord by
called retrograde transport. The rabies virus, for example, moves the vertebral column. Nerves can be damaged if the bones are
from a bite on the skin toward the brain or spinal cord where it infected at sites where the PNS penetrates this protective covering.
multiplies, ultimately killing the infected person. rabies, p. 658 Only rarely do infections in the bone surrounding the CNS spread
The brain and spinal cord (figure 26.1b) make up the central to the brain or spinal cord. Sometimes, however, they extend
nervous system (CNS). The brain is a very complex structure; through bone from the sinuses, mastoid air cells, or middle ear.
distinct parts have different functions. If an individual has a Skull fractures commonly predispose a person to recurring infec-
brain abscess, physicians can sometimes determine its location by tion of the CNS. mastoid air cells, p. 486 middle ear, p. 492

Central nervous system (CNS)

Brain is surrounded by meninges.
Encephalitis Peripheral nervous system (PNS)
Meningitis carries information to and from the CNS.
Rabies
African sleeping sickness
Spongiform encephalopathies
Ganglia

Dendrites
Synapse
Spinal cord
Meningitis
Poliomyelitis
Axon

Nerves
Leprosy
Botulism

Cell body
(a)

FIGURE 26.1 The Nervous System (a) Neurons receive

information via dendrites and transmit impulses over axons.
(b) The central nervous system is composed of the brain and
spinal cord. Peripheral nerves carry information to and from
the CNS; ganglia contain cell bodies of neurons. Nervous
system infections are shown in red.
? What part of a neuron is found in the peripheral nerves? (b)
 Part IV Infectious Diseases 643

(a)
Cerebrospinal fluid
reabsorbed into
bloodstream
Spinal cord
Subarachnoid
Subarachnoid Cerebrospinal space
space fluid formed
(b)

Pia Lumbar
mater vertebra
Dura mater
Venous
sinus Arachnoid
Cerebrospinal mater
Arachnoid fluid
mater Pia mater

(b)

Dura
mater
Ventricles
FIGURE 26.2 Meninges and Cerebrospinal
Fluid (a) Three layers of meninges—the dura
mater, arachnoid mater, and pia mater—surround
Spinal cord the CNS. CSF is formed in the ventricles, flows
within the subarachnoid space, and is reabsorbed
into the bloodstream. (b) A sample of cerebrospinal
fluid can be withdrawn and examined.
(a) ? What is the purpose of a spinal tap?

Deep inside the brain are four fluid-filled cavities called ven- structure, and a sample of CSF is withdrawn (figure 26.2b). The agent
tricles. The fluid within them, cerebrospinal fluid (CSF), is con- causing the infection can then be examined and cultivated for
tinually produced within the walls of ventricles, and spreads over identification. Other characteristics of the CSF are also examined,
the surface of the brain and spinal cord. CSF is reabsorbed into including glucose levels, protein levels, and white blood cell
the bloodstream at specialized sites within a venous sinus (figure counts. If bacteria are growing in CSF, glucose levels decrease
26.2a). CSF provides cushion and support for the brain and also and protein levels increase. Neutrophils also accumulate. Viral
transports nutrients and other materials throughout the CNS. meningitis is characterized by increased numbers of lymphocytes.
Three layers of membranes called meninges cover the surface The bloodstream is the chief source of CNS infections.
of the brain and spinal cord. The outer dura mater is tough and However, it is difficult for infectious agents to cross from the
provides a barrier to the spread of infection from bones surround- bloodstream to the brain because of the blood-brain barrier.
ing the central nervous system. It adheres closely to the skull This barrier depends on special cells lining capillaries in the CNS.
and vertebrae, and in some parts of the brain encloses a venous These cells, only a single layer thick, are so close to each other
sinus. The two inner membranes, the arachnoid mater and the pia that except for essential nutrients actively taken up, most mole-
mater, are separated by the subarachnoid space within which the cules cannot pass through them. The blood-brain barrier generally
cerebrospinal fluid flows (figure 26.2a). The innermost pia mater prevents pathogens from entering nervous tissue except when a
adheres to the nervous tissue. Inflammation or infection of these rare high concentration of the infectious agent circulates for a long
membranes is called meningitis. When both the meninges and the time in the bloodstream. Unfortunately, the barrier also prevents
brain are infected, the condition is meningoencephalitis. many medications, including penicillin, from crossing into the
The nervous system lies entirely within body tissues and has CNS unless their concentrations in the blood are very high.
no normal microbiota. CSF is generally sterile, so the presence of
microbes indicates an infection. To diagnose meningitis, a sample MicroByte
of CSF is obtained using a procedure called a spinal tap or lumbar Infections on the face can move through bone to veins on the brain
surface. A good rule is not to pop pimples on the upper part of
puncture. A needle is inserted into the subarachnoid space between
the face.
lumbar vertebrae where the spinal cord has tapered to a threadlike
644 Chapter 26 Nervous System Infections

MicroAssessment 26.1 Signs and Symptoms

Information within a neuron typically flows from dendrites to the
Bacterial meningitis usually begins like a mild cold. This is
cell body to the axon. Neurotransmitter molecules are released from followed by the sudden onset of a severe, throbbing headache;
the axon at a synapse where they bind to receptors on a neighboring fever; pain and stiffness of the neck and back; nausea; and vomit-
cell, initiating a response. The brain and spinal cord make up the ing. Deafness, confusion, loss of consciousness, and coma may
central nervous system (CNS). Nerves in the peripheral nervous develop. Death may occur rapidly due to inflammation and shock.
system (PNS) convey sensory and motor information to and from
the CNS. The CNS is protected by bone, by circulating cerebrospinal Causative Agent
fluid (CSF), and by membranes called meninges. Inflammation of the
meninges is termed meningitis. Infection can reach the CNS through Streptococcus pneumoniae, often called pneumococcus, is a
the bloodstream, by axonal transport, or by contact with infected Gram-positive lancet-shaped coccus, often seen in pairs (figure
bone. The blood-brain barrier prevents passage of most pathogens 26.3). Many strains are protected from phagocytosis by a polysac-
from the bloodstream into the CNS, but it also can prevent access to charide capsule. Antigenic differences in the capsules produce
the CNS by antimicrobial medications. over 90 recognized serotypes. When the pathogen is part of the
1. What vital molecules are carried from the cell body to the normal respiratory microbiota, the mucociliary escalator usually
synapse for release? prevents it from entering the lungs, where it could cause pneumo-
2. List the components of the CNS and of the PNS. nia or enter the bloodstream. mucociliary escalator, p. 337
3. Why can encephalitis usually be detected in CSF obtained from
the lower back? + Pathogenesis
Streptococcus pneumoniae is a common cause of otitis media,
sinusitis, and pneumonia, any of which can precede pneumococ-
26.2 ■ Bacterial Diseases cal meningitis. Encapsulated strains resist phagocytosis and may
of the Nervous System enter the bloodstream of an infected individual, bind to receptors
on meningeal cells, and pass into the cerebrospinal fluid, causing
Learning Outcomes meningitis. The damage associated with meningitis is largely due
4. Compare and contrast the various types of bacterial meningitis. to the inflammatory response. Fluids that accumulate in the area
cause brain swelling, and the clots that form in the capillaries can
5. Explain how Hansen’s disease can cause peripheral nerve damage.
block the blood supply, leading to tissue death. Inflammation can
6. Explain how botulism affects the nervous system.
also obstruct the normal outflow of cerebrospinal fluid, causing
the brain to be squeezed against the skull by the buildup of internal
Bacteria can infect peripheral neurons, but they most often infect
the meninges surrounding the central nervous system—causing
meningitis. Early symptoms are like a mild cold, but they can
quickly escalate. Bacterial meningitis can kill within hours, so
antibiotics are often given while the cause of the inflammation is
still being determined. The case-fatality rate for untreated bacte-
rial meningitis can approach 100%. Even with proper treatment,
the rate is still relatively high (about 10–20%). Survivors some-
times suffer permanent disabilities including deafness, blindness,
paralysis, or mental impairment.
The three most common species that cause meningitis—
Streptococcus pneumoniae, Neisseria meningitidis, and
Haemophilus influenzae—are often part of the normal microbiota
of the upper respiratory tract. They are transmitted through respi-
ratory droplets and routinely cause common diseases like sinusitis
or conjunctivitis. In a small percentage of infected people, how-
ever, the organisms invade the bloodstream and then spread to
the CNS. People with lowered immunity such as newborns and
the elderly are at most risk for meningitis. Young adults living
in crowded, confined environments such as military barracks
or college dormitories are also at increased risk. Streptococcus
pneumoniae, p. 497 conjunctivitis, p. 492 20 μm

FIGURE 26.3 Streptococcus pneumoniae (Pneumococci)

Pneumococcal Meningitis This Gram-stained smear shows large numbers of the Gram-positive
diplococci in the CSF of a person with pneumococcal meningitis;
Pneumococcal meningitis is caused by Streptococcus pneumoniae, the large red objects are polymorphonuclear leukocytes that have
part of the normal microbiota in the throat and nasopharynx of entered the spinal fluid in response to the infection.
many healthy individuals. Although best known as a cause of pneu- ? How would a Gram stain distinguish this organism from
monia, it is the leading cause of meningitis in adults. Neisseria meningitidis?
 Part IV Infectious Diseases 645

pressure. Mortality and neurological damage are more common

with this type of meningitis than with others. otitis media, p. 492
pneumococcal pneumonia, p. 496

Epidemiology
When the mucociliary escalator is impaired, perhaps by respira-
tory infections such as influenza, Streptococcus pneumoniae cells
can enter the lungs from the throat or nasopharynx, where they
are part of the normal microbiota. Bacteremia is seen in most
cases of pneumococcal meningitis, but trauma that allows contact
between the CSF and the nasopharynx can also cause infection.
The pathogen is spread by respiratory droplets, but very few who
are infected will develop meningitis. The case-fatality rate is about
20% in adults and much higher in the elderly. bacteremia, p. 384
FIGURE 26.4 Petechiae of Meningococcal Disease These
Treatment and Prevention skin lesions are characteristic of meningitis caused by Neisseria
Penicillin remains the drug of choice for susceptible strains of meningitidis.
Streptococcus pneumoniae, but some strains show antibiotic ? What causes petechiae?
resistance. For those cases of pneumococcal meningitis, a cepha-
losporin such as ceftriaxone or cefotaxime is administered, with
the possible addition of vancomycin.
Vaccines against S. pneumoniae offer protection from pneu-
monia and otitis media as well as meningitis, but are not effective most common in Africa and Asia. Like Neisseria gonorrhoeae,
against all of the different capsular antigens. A vaccine of the 23 N. meningitidis can vary some of its antigens and easily acquires
most common capsular antigens has been developed for use in DNA through horizontal gene transfer. serotype, p. 247 antigenic
vulnerable adults. This vaccine is not effective in children because variation, p. 390 horizontal gene transfer, p. 200
polysaccharide antigens are T-cell-independent and children under
two respond poorly to them. A different vaccine was specifically Pathogenesis
developed for this age group. It consists of seven of the capsular Meningococci inhaled in airborne droplets attach by pili to
antigens attached to a bacterial protein, forming an effective T-cell- mucous membranes and multiply. Specific proteins in their outer
dependent conjugate vaccine. Since the vaccine was licensed in membranes allow the bacterial cells to pass through epithelial
2000, childhood cases of pneumococcal meningitis caused by these cells lining the respiratory tract and into the bloodstream to the
seven strains have dropped significantly. However, incidence of meninges, although this is rare. Circulating Neisseria meningitidis
cases caused by other strains has increased. A new 13-valent pneu- release blebs of their outer membranes, and the endotoxin causes
mococcal conjugate vaccine was licensed in 2010; it will provide vasodilation and capillary leakage, leading to a drop in blood
protection against infections caused by six additional pneumococcal pressure. Endotoxic or septic shock results when blood pressure
serotypes. T-independent antigen, p. 367 conjugate vaccines, p. 424 becomes so low that circulation cannot adequately supply O2 to
vital body tissues. Capillary damage causes the petechiae associ-
ated with meningococcal meningitis. pili, p. 65 endotoxin, p. 394
Meningococcal Meningitis The capsule helps the cells avoid phagocytosis and protects
Neisseria meningitidis, often called meningococcus, is frequently them from the lethal actions of the complement system. Large
responsible for epidemics of meningitis. Although humans are numbers of neutrophils enter the CSF in response to the infection,
the only host, meningococcal meningitis is difficult to eliminate but the bacteria multiply faster than they can be destroyed. As
because N. meningitidis is commonly part of the normal respira- in pneumococcal meningitis, tissue damage and cerebral edema
tory microbiota of healthy individuals. often result from inflammation. complement system, p. 344

Signs and Symptoms Epidemiology

Symptoms of meningococcal meningitis are similar to those of Transmission of Neisseria meningitidis can occur when someone
pneumococcal meningitis but often include purplish spots on the is exposed to a person with the disease or to an asymptomatic car-
skin called petechiae (figure 26.4). Septic (endotoxic) shock can rier. Although most cases are sporadic, meningococcal meningitis
lead to rapid death. septic shock, pp.  394, 671 causes epidemics if it spreads via respiratory droplets through
crowded and stressed populations. Neisseria meningitidis can
Causative Agent cause meningitis at any age. As the incidence of cases in babies
Meningococcal meningitis is caused by Neisseria meningitidis, a has decreased due to immunization, a higher percentage of cases
Gram-negative encapsulated diplococcus. The most serious infec- is reported in young adults. In the United States, it is a relatively
tions are due to serotypes A, B, C, Y, and W135. Groups B and rare disease (fewer than 3 cases per 100,000 people annually).
C are most common in the United States and Europe; type A is The highest incidence worldwide occurs in the “meningitis belt”
646 Chapter 26 Nervous System Infections

extending between Senegal and Ethiopia in sub-Saharan Africa, Treatment and Prevention
where the population is about 300 million (figure 26.5). During If treated in time, meningococcal meningitis can usually be cured
the dry season between December and June, incidence may reach with penicillin or ceftriaxone. Most patients recover without
500 cases per 100,000 people. permanent nervous system damage. The case-fatality rate is less
than 10% in treated cases.
A vaccine composed of purified A, C, Y, and W135 capsular
polysaccharides has been available for years to control epidemics
and to immunize people at high risk. A conjugate vaccine was
approved in 2005 for the same four types, and an additional option
was licensed in 2010. A conjugate vaccine is recommended for all
11- to 18-year-olds, and for those 2 to 55 years of age who are at
increased risk, including college freshmen who will live in dormi-
tories. Group B strains possess a poorly immunogenic polysaccha-
ride, and no vaccine is yet available for their control. Antibacterial
Mauritania Mali Niger
medications, however, can be useful in controlling epidemics
Senegal Chad Eritrea in confined groups such as people in jails, nursing homes, and
Burkina Sudan
Faso schools. People intimately exposed to cases of meningococcal dis-
Guinea
Nigeria ease are routinely given prophylactic treatment with the antibiotic
Central Ethiopia
African Republic rifampin. Table 26.1 gives the main features of this disease.
Guinea-Bissau Benin Cameroon Uganda
Gambia Togo Democratic Kenya Haemophilus influenzae
Côte D'ivoire Ghana Republic
Of The Congo Meningitis
Haemophilus influenzae once caused meningitis in about 1 out of
FIGURE 26.5 The Meningitis Belt Meningitis is most prevalent 200 children under 5 years of age and was the leading cause of
in Mali, Burkina Faso, Niger, Chad, Sudan, Ethiopia, and northern meningitis in that age group. Hib vaccine, introduced in the late
Nigeria. 1980s, is a great success story as it has decreased incidence of this
? Why would the incidence of meningitis be seasonal in this region? type of meningitis in children by over 99%.

TABLE 26.1 Meningococcal Meningitis

1 Neisseria meningitidis inhaled, 4 Signs and Mild cold followed by headache, fever,
infects upper airways. 6 symptoms pain, stiff neck and back, vomiting,
2 1 petechiae
Bacteria enter the bloodstream
and are circulated throughout Incubation period 1 to 7 days
the body.
Causative agent Neisseria meningitidis, the
3 The bacteria damage meningococcus; a Gram-negative
skin capillaries and cause 7 diplococcus
petechiae.
Pathogenesis Meningococci adhere by pili, colonize
4 Bacteria infect the meninges 2 upper respiratory tract, enter
causing meningitis. 5 bloodstream; carried to meninges and
5 spinal fluid; inflammatory response
Lysing bacteria in the
obstructs normal outflow of fluid;
circulation release endotoxin,
increased pressure caused by obstructed
producing shock.
flow impairs brain function; damage
6 Inflammatory response in to motor nerves produces paralysis;
meninges can damage nerves endotoxin release causes shock.
of hearing, causing deafness,
and obstruct the flow of Epidemiology Close contact with a case or carrier;
inhalation of infectious droplets;
cerebrospinal fluid, causing
crowding and fatigue predispose to the
increased pressure inside the
disease.
brain.
3
7 Bacteria exit with respiratory Treatment Rifampin given to those exposed.
secretions. and prevention Penicillin, ceftriaxone for treatment.
Conjugate vaccine against serotypes A,
C, Y, and W135 used to immunize ages
2–55 years.
 Part IV Infectious Diseases 647

CASE PRESENTATION
The patient was a 31-month-old girl admitted 3. What age group is most susceptible to this antibodies are lacking and adaptive
to the hospital because of fever, headache, illness? immunity has not yet fully developed.
drowsiness, and vomiting. She had been previ- 4. Compare the pathogenesis of this disease Since 1987, vaccines consisting of type
ously well until 12 hours before admission, in children and adults. b capsular antigen conjugated with a pro-
when she developed a runny nose, malaise, tein, such as the outer membrane protein
and loss of appetite. Discussion of Neisseria meningitidis or diphtheria
Her birth and development were normal. 1. The patient had bacterial meningitis toxin, have been used to immunize infants
There was no history of head trauma. Her rou- caused by Haemophilus influenzae sero- and thus eliminate the immunity gap. As a
tine immunizations had been neglected. type b. The fact that she had been healthy result, meningitis caused by H. influenzae
On examination, her temperature was prior to her illness makes it unlikely that type b is now rare.
40°C (104°F), her neck was stiff, and she did other pathogens could be responsible. 4. Haemophilus influenzae type b strains are
not respond to verbal commands. 2. With treatment, the fatality rate is approx- referred to as “invasive” strains because
Her white blood cell count was elevated imately 5%. Formerly, ampicillin was they establish infection of the upper respi-
and showed a marked increase in the per- effective in most cases, but beginning in ratory tract, pass the epithelium, and enter
centage of polymorphonuclear neutrophils 1974, an increasing number of strains pos- the lymphatic vessels and bloodstream. In
(PMNs). Her blood sugar was in the normal sessed a plasmid coding for β-lactamase. this way, they gain access to the general
range. So far, however, these strains have been circulation and are carried to the central
A spinal tap was performed, yielding susceptible to the newer cephalosporin- nervous system. Most strains of H. influ-
cloudy cerebrospinal fluid under increased type antibiotics. Unfortunately, about one- enzae are non-invasive. Although they can
pressure. The fluid contained 18,000 white third of those who are treated and recover cause infections of respiratory epithelium,
blood cells per microliter (normally, there from the infection are left with permanent they usually do not enter the circulation.
are few or none), a markedly elevated protein damage to the nervous system, such as Most adults are immune to type b strains,
level, and a markedly low glucose level. Gram deafness or paralysis of facial nerves. but some adults develop meningitis from
stain of the fluid showed many tiny, Gram- Prompt diagnosis and correct choice non-invasive H.  influenzae strains that
negative coccobacilli, most of which were of antibacterial treatment minimize the gain access to the nervous system because
outside the white blood cells. chance of permanent damage. of a skull fracture or by direct extension
1. What is the diagnosis, and what is the 3. The peak incidence of this disease is to the meninges from an infected sinus or
causative agent? in the age range of 6 to 18 months, cor- middle ear.
2. What is the prognosis in this case? responding to the time when maternal

Signs and Symptoms and enter capillaries. The capsule helps them resist phagocytosis.
As with other types of meningitis, symptoms usually begin with Bacteremia can result in meningitis or other infections such as
a mild cold and progress to severe headache, fever, and vomiting. epiglottitis or cellulitis. Unencapsulated strains generally do not
Older children may report a stiff neck. Infants may show a bulging enter the bloodstream but can cause local infections such as sinus-
fontanelle (the gap between the bones of an infant’s skull). The itis and otitis media.
disease may progress rapidly to coma and death.
Epidemiology
Causative Agent Unlike meningococcal meningitis, Haemophilus influenzae usu-
Haemophilus influenzae, a Gram-negative non-motile rod, was ally does not cause epidemics. Healthy adult carriers are often
named because it was thought to have caused an influenza epi- the source of sporadic infection by H. influenzae type b. Children
demic in the late 1890s. Although it did not cause the epidemic, rarely carry Hib due to the high rate of immunization. The patho-
it was and is often isolated from influenza patients. Encapsulated gen is spread by respiratory droplets, but it rarely causes disease
strains labeled a through f cause most disease in children; bacteria except in unimmunized infants. Case-fatality rate when untreated
with capsular antigens of type b, abbreviated Hib, cause the most is about 90%. Even with treatment, about 5% die and 10–30% of
serious disease. Unencapsulated strains are part of the normal children have lasting neurological damage.
microbiota of the nasopharynx in many healthy individuals, and
some people also carry Hib. In 1991, H. influenzae earned the Treatment and Prevention
distinction of being the first free-living organism to have its small Treatment with cefotaxime or ceftriaxone is usually successful
genome completely sequenced. against Haemophilus influenzae. People who have been in close
contact with an infected individual may be given rifampin to help
Pathogenesis prevent the disease.
Encapsulated Haemophilus influenzae in the upper respiratory The conjugate Hib vaccine contains the type b polysaccharide
tract can bind to and penetrate the epithelium with the aid of pili antigen attached to a protein such as diphtheria toxoid. The protein
648 Chapter 26 Nervous System Infections

Other cases are caused by Listeria monocytogenes from the blood-

stream of an infected mother. Lancefield grouping, p. 487
25
Pathogenesis
20 Infection of the meninges is usually preceded by bacteremia in
the newborn. Inflammation increases intracranial pressure that
can block CSF flow, causing hydrocephalus. Disruption of blood
15 flow may damage nervous tissue. Infection may also lead to
Incidence

brain abscess.
10
Epidemiology
Neonates usually acquire these infections from the mother’s
5 genital tract shortly before or during birth. Premature or low birth
weight babies are at most risk for meningitis. Neonatal meningitis
causes death in 6–20% of affected newborns. Those who survive
0
1990 1992 1994 1996 1998 2000 2002 2010 often have long-lasting consequences such as hearing loss or
Year mental disability.

FIGURE 26.6 Rate of Serious Haemophilus influenzae Treatment and Prevention

Disease per 100,000 Children Less than Age 5, United States, Treatment of neonatal meningitis includes intravenous dosage
1990–2010 Before the availability of conjugate vaccines in late
1987, H. influenzae type b was the most common cause of bacterial with a mixture of antibacterial drugs such as ampicillin and gen-
meningitis in preschool children. tamicin effective against both group B streptococci and E. coli.
? What are the components of the conjugate Hib vaccine?
Other drugs may be added after the causative agent is identified.
The Centers for Disease Control and Prevention (CDC)
recommends that the vagina and rectum of pregnant women be
tested for group B streptococci late in pregnancy. Women with
component makes the vaccine effective in children under age two, positive cultures can then be treated with an appropriate antibac-
who otherwise respond poorly to polysaccharide antigens. Infants terial medication shortly before or during labor. Screening and
in the United States are routinely vaccinated against Hib begin- subsequent treatment can decrease the incidence of serious group
ning at 2 months of age (figure 26.6). This vaccine is relatively B streptococcal disease by more than 75%.
expensive and must be administered more than once, making
it less practical in developing countries, where Hib infections
are still responsible for most deaths due to bacterial meningitis.
Listeriosis
conjugate vaccines, p. 424 Meningitis is the most common result of listeriosis, a foodborne
disease caused by Listeria monocytogenes. This organism gener-
ally causes only a small percentage of meningitis cases in the
Neonatal Meningitis United States, but epidemics may occur.
Most cases of neonatal meningitis (during the first month of life)
are caused by bacteria that colonize the mother’s birth canal. The Signs and Symptoms
common causes of meningitis in adults (Neisseria meningitidis, Listeria monocytogenes infections are generally asymptomatic or
Streptococcus pneumoniae, and Haemophilus influenzae) seldom mild in most healthy people. Symptomatic listeriosis is usually
cause meningitis in newborns because most mothers have antibod- characterized by fever and muscle aches, and sometimes nausea or
ies against them. These antibodies cross the placenta and protect diarrhea. Most of the cases requiring medical attention have men-
the baby until it is about 6 months old. ingitis with fever, headache, stiff neck, and vomiting. Pregnant
women who become infected often miscarry or deliver terminally
Signs and Symptoms ill premature or full-term infants.
Meningitis symptoms in an infant may be vague, as it is difficult
to tell if a baby has a headache or a stiff neck. Signs of meningitis Causative Agent
in a newborn may include fever and vomiting, irritability, poor Listeria monocytogenes is a motile, non-spore-forming, faculta-
feeding, lethargy, and bulging fontanels. tively anaerobic, Gram-positive rod that can grow at 4°C. The
organism can grow in refrigerated foods even if vacuum-packaged.
Causative Agents
The most common cause of meningitis in newborn infants is Pathogenesis
Streptococcus agalactiae, a Lancefield group B streptococcus The mode of entry in sporadic cases of listeriosis is usually unclear,
that colonizes the vagina of many women. Certain Gram-negative but during epidemics it is generally via the gastrointestinal tract.
rods, such as encapsulated strains of E.  coli, originate from the Gastrointestinal symptoms may or may not occur, but the bacteria
mother’s intestinal tract and also can cause neonatal meningitis. promptly penetrate the intestinal mucosa—through the M cells
 Part IV Infectious Diseases 649

and into the Peyer’s patches—and then enter the bloodstream. The
resulting bacteremia is the source of meningeal infection. In preg-
nant women, L. monocytogenes crosses the placenta and produces
widespread abscesses in tissues of the fetus. Babies infected at the
time of birth usually develop meningitis after an incubation period
of 1 to 4 weeks. M cells, p. 358 Peyer’s patches, p. 358

Epidemiology
Listeria monocytogenes is widespread in natural waters and
vegetation, and can be carried in the intestines of asymptomatic
humans and other animals. Pregnant women, the elderly, and those
with underlying illnesses such as immunodeficiency, diabetes,
cancer, and liver disease are especially susceptible to listeriosis.
Epidemics have resulted from L. monocytogenes contaminating
foods including coleslaw, non-pasteurized milk, pork tongue in
jelly, some soft cheeses, and hot dogs. Because the organisms can
grow in commercially prepared food stored at refrigeration tem-
peratures, thousands of infections have originated from a single
food-processing plant.

Treatment and Prevention

Most strains of L. monocytogenes remain susceptible to antibac-
terial medications such as penicillin. Even though the disease is
often mild in pregnant women, prompt diagnosis and treatment are
important to protect the fetus.
Listeria monocytogenes can be killed by thoroughly cook-
ing poultry, pork, beef, and other meats. To reduce the risk of
cross-contamination, uncooked meats should not be kept with FIGURE 26.7 Bacteriophage in Food Safety When sprayed
other foods; countertops and utensils should be cleaned after onto ready-to-eat foods, bacteriophage preparations may reduce the
food preparation; and raw vegetables should be thoroughly risk of listeriosis.
washed before eating. Pregnant women and others at high risk ? What is the source of most listeriosis infections?
are advised to avoid soft cheeses, refrigerated meat spreads, and
smoked seafood. They should also heat cold cuts and hot dogs
before eating them and avoid the fluids that may be in the pack- its presence is recorded on the food label. Use of this bacterio-
aging. In 2006, the U.S. Food and Drug Administration approved phage additive has increased the safety of these foods. The main
a food additive consisting of a mixture of bacteriophage strains causes of acute bacterial meningitis are compared in table 26.2.
that lyse L.  monocytogenes (figure 26.7). The additive can be Some of the main features of listeriosis are presented in
sprayed on a variety of meats during the production process, and table 26.3. bacteriophages, p. 304

TABLE 26.2 Main Causes of Acute Bacterial Meningitis Compared

Agent Characteristics Source Vaccine? Age Group Usually Affected

S. pneumoniae Gram-positive diplococci Respiratory system Yes, multiple Mostly late teens and adults
types
N. meningitidis Gram-negative diplococci Respiratory system Yes, four Mostly ages 2–20 years; can cause epidemics
types
H. influenzae Gram-negative coccobacilli Respiratory system Yes, type b Young children

S. agalactiae Gram-positive cocci in chains Colon, vagina No Mostly neonates; others with underlying diseases

E. coli Gram-negative rods; usually Colon No Mostly neonates

a specific encapsulated type
L. monocytogenes Gram-positive rods; multiply Environment; No Pregnant women, neonates; elderly
at refrigerator temperatures contaminated cheeses,
cold cuts, other foods
650 Chapter 26 Nervous System Infections

TABLE 26.3 Listeriosis

1 Causative organism Listeria Signs and Fever and muscle aches, with or without
monocytogenes is ingested symptoms gastrointestinal symptoms; headache
4
with food such as soft cheeses, and stiff neck mark the onset of
non-pasteurized milk, hot meningitis
dogs, or smoked fish. Incubation
1 A few days to 2 to 3 months; in
2 The bacteria rapidly penetrate period newborn babies, 1 to 4 weeks
the intestinal epithelium
Causative Listeria monocytogenes, a
and establish bacteremia,
agent non-spore-forming Gram-positive
especially in pregnant
rod able to grow at 4°C
women, the elderly, and the
immunodeficient. Pathogenesis Ingested L. monocytogenes cells
2 penetrate the intestinal epithelium and
3 In pregnant women,
enter the bloodstream; the resulting
circulating L. monocytogenes
bacteremia spreads to the meninges,
cross the placenta and fatally
3
causing meningitis.
infect the fetus; bacteria
transmitted to the baby at Epidemiology Epidemics from contaminated soft
2
birth cause meningitis in 1 to cheeses, non-pasteurized milk,
4 weeks. The mother usually coleslaw, hot dogs. Pregnant women
does not have a serious illness. get bacteremia, fetal infection,
miscarriage. Infants contract infection
4 In older people and those
at birth, develop meningitis in the first
with underlying diseases,
month of life. Elderly and those with
L. monocytogenes attacks
immunodeficiency, cancer, and diabetes
brain and meninges, causing
also at high risk.
meningitis, brain abscesses.
Treatment Antibacterial medications such as
and prevention penicillin, given promptly, are effective
treatment. Care in handling, cooking
of raw meats; thorough washing of
vegetables; reheating of cold cuts, hot
dogs, and refrigerated leftovers.

Hansen’s Disease (Leprosy) Causative Agent

Hansen’s disease, also known as leprosy, is an ancient dis- Mycobacterium leprae is aerobic, rod-shaped, and acid-fast
ease appearing throughout written history from about 600 B.C. (figure  26.9). It grows very slowly with a generation time of
Disfigurement, loss of limbs, and blindness can result from skin about 12 days, and prefers the slightly cooler temperatures of the
and peripheral nerve involvement. The number of new cases of body’s extremities. Hansen’s disease is usually diagnosed based
Hansen’s disease has decreased dramatically since the introduc- on clinical findings, but skin biopsies that show acid-fast rods
tion of multidrug therapy; today it is most often seen in tropical can provide an early indication of nerve invasion. Despite many
or developing countries, such as India, Mozambique, and Brazil. attempts, M. leprae has not been grown in the absence of living
Still, millions of people continue to suffer from the residual effects cells. It can, however, grow in the footpads of mice, in armadillos,
of their disease and the stigma associated with it. In the United and in mangabey monkeys. Moreover, a gene library of M. leprae
States, about 200 cases are reported annually. This number has has been made and expressed in E.  coli, providing large quanti-
increased in recent years, mostly in states receiving immigrants ties of the organism’s antigens for study. acid-fast stain, p.  48
Mycobacterium, p. 264 DNA library, p. 221
from countries where the disease is endemic.

Signs and Symptoms Pathogenesis

Hansen’s disease begins gradually, usually with the onset of Mycobacterium leprae is the only known human pathogen that
pigmentation changes and increased or decreased sensation in preferentially infects peripheral nerves. From there, the course
certain areas of skin. These areas may thicken—losing hair, sweat of the infection depends on the immune response of the host. In
glands, and all sensation. The nerves of the arms and legs may most cases, cell-mediated immunity develops against the invad-
become visibly enlarged with accompanying pain, later changing ing bacteria, and activated macrophages limit their spread. The
to numbness. Loss of nerve activity can lead to muscle wasting, chronically infected nerve cells, however, are progressively dam-
ulceration, and finally loss of fingers or toes due to unnoticed or aged by attacking immune cells, leading to disabling deformities,
untreated injury (figure 26.8). In more severe cases, changes are resorption of bone, and skin ulcerations that characterize the
most obvious in the face, with thickening of the nose and ears disease. The disease often spontaneously stops progressing, and
and deep wrinkling of the facial skin. Collapse of the supporting the nerve damage, although permanent, does not worsen. This lim-
structure of the nose leads to congestion and bleeding. ited type of Hansen’s disease, in which cell-mediated immunity
 Part IV Infectious Diseases 651

FIGURE 26.8 Effects of

Leprosy (a) An early symptom
of leprosy is a change in skin
pigmentation. (b) Notice the absence
of fingers and sunken nose as a
result of severe disease.
? Why are the fingers, toes, and
nose often affected by leprosy?

(a) (b)

successfully stops the proliferating bacteria, is called tuberculoid Epidemiology

leprosy (also called paucibacillary Hansen’s disease). People with Transmission of Mycobacterium leprae is by direct human-to-
tuberculoid leprosy rarely, if ever, transmit the disease to others. human contact. The source of the organisms is mainly nasal
cell-mediated immunity, p. 356 macrophage, pp. 340, 348 secretions of lepromatous cases, which can transport M. leprae to
When cell-mediated immunity to M. leprae fails to develop or mucous membranes or skin abrasions of other individuals. Even
is suppressed, unrestricted growth of M. leprae occurs, leading to a then, the disease develops in only a tiny minority, being controlled
form of Hansen’s disease called lepromatous leprosy (also called by immune defenses in the rest. Natural infections with M. leprae
multibacillary Hansen’s disease). The bacteria first multiply in the also occur in wild nine-banded armadillos. Although armadillos
cooler tissues of the body, notably in skin macrophages and peripheral have not been considered an important source of human leprosy,
nerves, and later throughout the body. The tissues and mucous mem- a recent study suggests that contact with the animals may indeed
branes contain billions of M. leprae, but there is almost no inflam- transmit the disease.
matory response to them. Mucus of the nose and throat contains high Eradication of Hansen’s disease, defined as less than 1 case
numbers of the pathogen, which can easily be transmitted to others. per 10,000 individuals, has been achieved in most countries where
the disease was endemic. It is difficult to completely eradicate,
however, due to the long generation time of M. leprae. The long
generation time results in an incubation period of about 3 years
(range, 3 months to 20 years), during which time the disease can
remain undetected. generation time, p. 83

MicroByte
During much of history, Hansen’s disease was so feared that
“lepers” were forced to carry a bell or horn to warn others of
their presence.

Treatment and Prevention

Early treatment can keep Hansen’s disease from progressing.
Since 1995, the World Health Organization has provided free
multidrug therapy to patients, radically reducing the number
M. leprae 10 μm
of cases. Tuberculoid leprosy can be successfully treated by a
FIGURE 26.9 Mycobacterium leprae in a Biopsy Specimen combination of dapsone and rifampin administered for 6 months.
The red areas are dense masses composed of millions of the bacteria, Lepromatous leprosy is generally treated for a minimum of 2 years,
a typical finding in lepromatous leprosy. with addition of a third drug, clofazimine, to the treatment regimen.
? In what human cells would you expect to find these bacteria? Multiple drug therapy is required to prevent drug-resistant strains
652 Chapter 26 Nervous System Infections

TABLE 26.4 Hansen’s Disease (Leprosy) Causative Agent

Botulism is caused by the strictly anaerobic, Gram-positive, spore-
Signs and Skin lesions that lack sensation, deformed forming, rod-shaped bacterium Clostridium botulinum (figure
symptoms face, loss of fingers or toes
26.10). The endospores generally survive boiling and can then
Incubation period 3 months to 20 years; usually 3 years germinate to form vegetative cells if the environment is favor-
Causative agent Mycobacterium leprae, an acid-fast rod able (nutrient-rich, anaerobic conditions, a pH above 4.5, and
that has not been grown in the absence of a temperature above 4°C). Several types of a neurotoxin called
living cells botulinum toxin are produced by different strains of C. botulinum,
Pathogenesis Invasion of small nerves of skin; and all cause paralysis. Types A, B, and E are responsible for most
multiplication in macrophages; course human cases. neurotoxin, p. 392
of disease depends on immune response
of host; activated macrophages limit
growth of bacterium; attack of immune Pathogenesis
cells against infected nerve cells produces Vegetative cells of Clostridium botulinum growing in food release
nerve damage, leading to deformity; in botulinum toxin, one of the most powerful poisons known. A few
lepromatous leprosy, lymphocytes fail to
react to the bacteria, allowing unrestrained
milligrams could kill the entire population of a large city. Indeed,
growth of M. leprae. cases of botulism have resulted from eating a single, contaminated
Epidemiology Direct contact with M. leprae from mucous
string bean, or from licking a finger when tasting a food contami-
membrane secretions nated with botulinum toxin. When a person eats the contaminated
Treatment Treatment: dapsone plus rifampin for
food, the toxin passes through the stomach and into the small
and prevention months or years; clofazimine added for intestine. Once absorbed into the bloodstream, the toxin can cir-
lepromatous disease. No vaccine. culate for 3 weeks or more.
Circulating botulinum toxin attaches to motor neurons, block-
ing transmission of signals to the muscles, producing paralysis.
Botulinum toxin is an A-B toxin. The B portion binds to specific
from developing. No proven vaccine to control Hansen’s disease
receptors on motor nerve endings, and the A portion enters the
is yet available. Some features of Hansen’s disease are summa-
nerve cell, where it inactivates proteins that regulate the release of
rized in table 26.4.
the neurotransmitter. Unlike tetanus toxin, which blocks inhibitory
neurons and results in spastic muscle contraction, botulinum toxin
Botulism blocks muscle contraction resulting in flaccid paralysis. Because
Botulism is not a nervous system infection, but a severe form it blocks nerve transmission to muscles, botulinum toxin can be
of intoxication, usually foodborne. The name “botulism” comes used to treat people with certain chronic, spastic conditions such
from botulus, meaning “sausages,” chosen because some of the as crossed eyes. Commercially available botulinum toxin, or
earliest recognized cases occurred in people who ate contaminated Botox, is used in a cosmetic treatment that relaxes facial muscles,
sausages. Like other clostridia, Clostridium botulinum produces
heat-resistant endospores. The endospores themselves do not
cause disease, but they germinate in nutrient-rich anaerobic envi-
ronments and produce a powerful exotoxin that causes the charac-
teristic symptoms. Botulism is considered in this chapter because
its key symptom is paralysis.
On a global basis, foodborne botulism is the most common
type of this disease. It typically occurs when inadequate canning
practices fail to destroy all endospores, and those surviving germi-
nate and grow in the food. Some botulism cases occur when endo-
spores are ingested and then germinate and colonize the intestine
(intestinal botulism). Others result from endospore contamination
of a wound where they then germinate and multiply (wound botu-
lism). foodborne botulism, p. 757

Signs and Symptoms

Symptoms of foodborne botulism usually begin 12 to 36 hours
after ingesting toxin-contaminated food. Most cases begin with
Endospores 10 μm
dizziness, dry mouth, and blurred or double vision, indicating eye
muscle weakness. Abdominal symptoms, including pain, nausea, FIGURE 26.10 Clostridium botulinum Notice the lighter-colored
vomiting, and diarrhea or constipation can also occur. Progressive endospores in this Gram stain. These are not reliably killed by the
paralysis generally involves all voluntary muscles; respiratory temperature of boiling water.
paralysis is the most common cause of death. ? What is an endospore?
 Part IV Infectious Diseases 653

resulting in fewer wrinkles (see Perspective 31.1). A-B toxin, p. 392 Treatment and Prevention
neurotransmitter, p. 642 tetanus, p. 555 Foodborne botulism is treated by administering antitoxin intrave-
Intestinal botulism occurs when C. botulinum colonizes the nously as soon as possible after the diagnosis. The antitoxin, how-
intestine. This primarily happens in infants, which is why it is ever, only neutralizes toxin circulating in the bloodstream. The
also known as infant botulism. When C. botulinum endospores nerves already affected recover slowly, over weeks or months.
are ingested, they can germinate, and the resulting vegetative cells Artificial respiration with a mechanical ventilator may be required
may grow in the intestine, where they produce toxin. This rarely for prolonged periods until affected nerve endings can regener-
happens in adults, presumably because the normal intestinal micro- ate. Most intestinal botulism patients recover without receiving
biota successfully compete with the germinating C. botulinum antitoxin treatment, although respiratory support and tube feeding
cells. Exceptions occur, however, particularly in immunodeficient may be needed until the pathogens are replaced by normal intes-
patients whose normal microbiota has been suppressed by anti- tinal microbiota. For wound botulism, surgical removal of dirt
biotic treatment. Intestinal botulism is characterized by constipa- and dead tissue helps to eliminate any organisms and unabsorbed
tion followed by generalized paralysis that can range from mild toxin. Enemas and gastric washing are used to do the same for
lethargy to respiratory insufficiency. intestinal botulism.
Clostridium botulinum can also colonize dirty wounds, espe- Prevention of foodborne botulism depends on proper steril-
cially those containing dead tissue. The bacteria do not invade, but ization and sealing of food at the time of canning. Contamination
they can multiply in dead tissue. Botulinum toxin then diffuses does not always result in a spoiled smell, taste, or appearance,
into the bloodstream. but fortunately, the toxin is heat-labile (destroyed by heat). Heat-
ing home-canned, low-acid foods (pH over 4.5) to 100°C for
Epidemiology 15 minutes just prior to serving should ensure that they are safe.
Clostridium botulinum endospores are widely distributed in Immunity does not develop in response to the low levels of toxin,
soils and aquatic sediments worldwide. Still, fewer than 30 so a person may get botulism more than once. The main features
cases of foodborne botulism per year are typically seen in the of botulism are presented in table 26.5.
United States. Most of these are from eating preserved fish or
improperly home-canned foods. Strict controls on commercial
TABLE 26.5 Botulism
canners have drastically reduced the number of botulism out-
breaks. Today, in the United States, intestinal (infant) botulism Signs and Blurred or double vision, weakness, nausea,
is more common than foodborne botulism, but there are gener- symptoms vomiting, diarrhea; generalized paralysis
ally fewer than 100 cases per year. Ingestion of honey has been and respiratory insufficiency
implicated in infant botulism cases, leading to the recommenda- Incubation period Usually 12 to 36 hours
tion that honey—a source of C. botulinum spores—not be given Causative agent Clostridium botulinum, an anaerobic,
to children under 2 years old. Most cases of wound botulism in Gram-positive, spore-forming, rod-shaped
the United States are due to wounds caused by abuse of injected bacterium
drugs (figure 26.11). Pathogenesis Clostridium botulinum endospores
germinate in food and release neurotoxin.
Toxin is ingested, absorbed, and is carried
by the bloodstream to motor nerves; toxin
acts by blocking the transmission of nerve
Foodborne signals to the muscles, producing paralysis;
110 Infant C. botulinum can also colonize intestines or
Wound wounds and cause generalized weakness or
100
paralysis.
90
Epidemiology Ingestion of toxin produced by vegetative
80
Clostridium botulinum cells growing in
Number of cases

70 a food, often an improperly processed

home-canned, low-acid food. Endospores
60
widespread in soil, aquatic sediments, and
50 Baked potatoes (TX) dust. Organisms can colonize the intestines
40 of adults and infants with deficiencies in
normal microbiota, and wounds containing
30 dirt and dead tissue, including those caused
20 by injected-drug abuse.
10 Fermented fish/ Treatment Treatment: enemas and stomach washing
seafood (AK) Chili sauce (TX)
and prevention to remove toxin, cleaning infected wounds
0
of dirt and dead tissue, intravenous
1990 1995 2000 2005 2009
Year administration of antitoxin, and artificial
respiration. Education in proper home-
canning methods; heating food to boiling
FIGURE 26.11 Botulism in the United States,1987–2009 for 15 minutes just prior to serving.
? What is the most common form of botulism in the United States?
654 Chapter 26 Nervous System Infections

MicroAssessment 26.2 cases of viral meningitis. Of these, the most common offenders
are coxsackie viruses, which can cause throat or chest pain, and
Bacterial infections of the nervous system are uncommon, but have echoviruses, which can cause a rash. picornaviruses, p. 308
a relatively high case-fatality rate. Bacteria commonly carried in
the upper respiratory tract (Streptococcus pneumoniae, Neisseria
meningitidis, and Haemophilus influenzae) cause most cases of Pathogenesis
bacterial meningitis. Organisms from the mother’s birth canal are Enteroviruses characteristically infect the throat, intestinal epi-
frequent causes in infants less than 1 month old. Meningitis is a
thelium, and lymphoid tissue, and then spread to the bloodstream
common complication of listeriosis, usually a foodborne disease
caused by Listeria monocytogenes. Hansen’s disease (leprosy) is causing viremia. This can result in meningeal infection. The
caused by Mycobacterium leprae, which preferentially invades inflammatory response differs from bacterial meningitis in that
peripheral neurons. Botulism is a toxin-mediated disease characterized fewer cells usually enter the cerebrospinal fluid, and more are
by flaccid paralysis that is caused by Clostridium botulinum. monocytes rather than neutrophils. Viral meningitis is typically
4. Why are most newborn babies unlikely to contract less severe than bacterial meningitis and causes little lasting neu-
pneumococcal, meningococcal, or Haemophilus meningitis? rological damage. viremia, p. 384

5. Differentiate between the two types of Hansen’s disease.

6. Why are so many infant botulism cases associated with Epidemiology
honey? + Enteroviruses are relatively stable in the environment, some-
times even persisting in chlorinated swimming pools. Enteroviral
meningitis is transmitted by the fecal-oral route, unlike bacterial
26.3 ■ Viral Diseases of meningitis that is usually transmitted by respiratory droplets.
The feces of infected individuals often contain viruses for
the Nervous System weeks.
Learning Outcomes
Treatment and Prevention
7. Compare and contrast viral meningitis with viral encephalitis.
No specific treatment is available. Handwashing and avoidance
8. Explain the history of poliomyelitis and why it has been targeted
for global elimination. of crowded swimming pools are reasonable preventive measures
when cases of aseptic meningitis are present. There are no vac-
9. Outline the steps in the pathogenesis of rabies in wild animals and
explain how it can be prevented in humans. cines against coxsackie and echoviruses. The main features of
viral meningitis are presented in table 26.6.
A wide variety of viruses can infect the nervous system causing
meningitis or encephalitis. Poliomyelitis, now targeted for global
elimination, and rabies, a disease normally found in animals,
result from viral infection of neurons. Many viruses that typically
TABLE 26.6 Viral Meningitis
affect other body systems can occasionally infect the CNS. These
include the Epstein-Barr virus; the mumps, rubeola, varicella- Signs and Abrupt onset, fever, severe headache, stiff
zoster, and herpes simplex viruses; and, most commonly, human symptoms neck, often vomiting; sometimes sore throat,
enteroviruses. large parotid glands, rash, or chest pain
Incubation period Usually 1 to 2 weeks for enteroviruses, 2 to 4
weeks for mumps
Viral Meningitis Causative agents Most cases: small non-enveloped RNA
Viral meningitis is more common and much milder than bacterial enteroviruses of the picornavirus family,
meningitis. It usually does not require specific treatment, and indi- usually coxsackie or echoviruses. Mumps
virus common in unimmunized populations.
viduals generally recover in 7 to 10 days. Viruses are responsible
for most cases of “aseptic meningitis,” meaning that a lumbar Pathogenesis Viremia from primary infection spreads
to the meninges. Fewer leukocytes enter
puncture reveals no microorganisms in the cerebrospinal fluid.
cerebrospinal fluid than with bacterial
infections, and many are mononuclear,
Signs and Symptoms usually no decrease in CSF glucose.
The onset of viral meningitis is typically abrupt, with fever and Epidemiology Enteroviruses transmitted by the fecal-oral
severe headache above or behind the eyes, sensitivity to light, route, mumps by respiratory secretions and
and a stiff neck. Nausea and vomiting are common. In addition, saliva. Enteroviruses transmission mainly
summer and early fall; mumps in fall and
depending on the causative agent, there may be a sore throat, chest
winter.
pain, swollen parotid salivary glands, or a skin rash.
Treatment No specific treatment. Handwashing,
and prevention avoiding crowded swimming pools during
Causative Agents enterovirus epidemics; mumps vaccine for
Non-enveloped RNA viruses—members of the enterovirus sub- mumps prevention.
group of picornaviruses—are responsible for at least half of the
 Part IV Infectious Diseases 655

Viral Encephalitis bloodstream that mosquitoes can transmit the virus from one host
to another. Humans are an accidental dead-end host—they can
Although viral meningitis is usually mild, viral encephalitis is
acquire the virus from a mosquito bite but do not develop suffi-
more likely to cause death or permanent disability. Viral encepha-
cient viremia to transmit it back to the arthropod vector.
litis can be sporadic, resulting in a few widely scattered cases
The LaCrosse virus infects Aedes mosquitoes, which pass it
occurring routinely, or it can be epidemic. Sporadic encephalitis
directly from one mosquito to another in semen. They feed on and
is usually due to activation of latent herpes simplex viruses. Most
infect squirrels and chipmunks, where the virus is amplified and
people recover from the disease but are left with permanent dam-
spread to uninfected mosquitoes feeding on the rodents’ blood.
age such as epilepsy, paralysis, deafness, or mental impairment.
The West Nile virus, introduced into New York from the
herpes simplex viruses, p. 582
Middle East in the summer of 1999, has a bird reservoir. Migrating
Signs and Symptoms birds quickly spread the virus across the country (figure 26.12).
Some arboviruses have a broad host range and cause disease
The onset of viral encephalitis is usually abrupt, with fever, head-
in horses as well as humans. This gives rise to the names eastern
ache, and vomiting, as well as possible disorientation, localized
and western equine encephalitis.
paralysis, deafness, seizures, or coma.

Causative Agents Treatment and Prevention

Epidemic viral encephalitis is usually caused by arboviruses There is no proven antiviral therapy for epidemic viral encephalitis.
(arthropod-borne viruses), a group of enveloped, single-stranded The blood of sentinel chickens in cages with free access to mos-
RNA viruses transmitted by insects, mites, or ticks. The leading quitoes is tested periodically for evidence of arbovirus infection. A
causes of epidemic encephalitis in the United States are all trans- positive test would trigger an encephalitis alert, like this St. Louis
mitted by mosquitoes. These include LaCrosse encephalitis virus, encephalitis alert issued in Florida: “Avoid outdoor activity during
St. Louis and West Nile encephalitis viruses, and eastern and west- evening and night, the peak hours of biting for the Culex mosquito
ern equine encephalitis viruses (table 26.7). arthropods, p. 298 vector. If outdoors, wear long sleeves and pants. Make sure win-
dows and porches are properly screened. Use insect repellents and
Pathogenesis insecticides.” Equine encephalitis viruses generally infect horses
Viruses multiply at the site of a mosquito bite and in local lymph 1 or 2 weeks before the first human cases appear, so these cases pro-
nodes, producing mild and brief viremia. Relatively few infected vide a warning to increase protection against mosquitoes. A vaccine
individuals develop encephalitis. Some develop viral men- against eastern equine encephalitis is approved for use in horses
ingitis, or mild fever and headache. If viruses infect cells
of the blood-brain barrier, they can enter the brain and
replicate in neurons, causing destruction of brain tis-
sue. Neutralizing antibodies stop disease progression.
Case-fatality rates range from about 2% with LaCrosse
encephalitis to 35–50% with eastern equine encephali-
tis. Emotional instability, epilepsy, blindness, or paral-
ysis may occur in 5–50% of those who recover. The
likelihood of disability depends largely on the kind of
virus and the age of the patient. As with many diseases,
the very young and the elderly are most affected.

Epidemiology
The common types of epidemic viral encephalitis are all
zoonoses with a natural reservoir in birds or other small ani-
mals. Natural hosts can develop such high levels of virus in the

Arboviruses That Cause

TABLE 26.7 Encephalitis in the United States Per 100,000 population
0.00
Virus Family 0.01–0.24
0.25–0.49
LaCrosse encephalitis virus Bunyavirus 0.50–0.99
≥1.00
St. Louis encephalitis virus Flavivirus
West Nile encephalitis virus Flavivirus
Eastern equine encephalitis virus Togavirus FIGURE 26.12 Areas Reporting West Nile Virus Infection,
Western equine encephalitis virus Togavirus United States, 2010
? What is an arbovirus?
656 Chapter 26 Nervous System Infections

TABLE 26.8 Epidemic Viral Encephalitis

1 Infected mosquito introduces 3 Signs and Abrupt onset, fever, headache,

encephalitis virus. 4 symptoms vomiting, disorientation, paralysis,
2 seizures, deafness, coma
Virus multiplies locally,
establishes brief low-level Incubation period First symptoms within a few days;
viremia. encephalitic symptoms often within
3 Virus crosses blood-brain 2 the first week
barrier and preferentially Causative agent One of five arboviruses, LaCrosse,
attacks the brain. St. Louis, West Nile, western equine,
2
4 Destruction of brain tissue or eastern equine
causes death or permanent Pathogenesis Replication of virus at the site of
disabilities such as emotional 1 the mosquito bite, replication in
instability, mental disability, lymph nodes, then viremia invasion
paralysis of face, arm, leg. of brain tissue. Nerve cells in the
4
5 Due to brief viremia, there brain destroyed. Process halted by
is no exit for the virus, thus neutralizing antibody.
humans are the final host.
Epidemiology Viruses transmitted to humans from
birds or rodents by mosquitoes.
4
Treatment No accepted treatment for arboviral
and prevention encephalitis. Chicken sentinels
to warn of arbovirus epidemics.
Insecticides and other anti-mosquito
preventive measures.

and has also been used to protect the emu—a large, domesticated, syndrome is probably due to the death of these nerve cells after
meat-producing bird susceptible to this virus. The main features of doing double duty for many years.
epidemic viral encephalitis are presented in table 26.8.
Causative Agent
Poliomyelitis Poliomyelitis is caused by three types of polioviruses—
The characteristic feature of poliomyelitis (also called polio, or designated 1, 2, and 3—distinguished using different antisera.
infantile paralysis) is destruction of motor neurons resulting in These non-enveloped, single-stranded RNA viruses are members
paralysis of a group of muscles, such as those of an arm or leg. of the enterovirus subgroup of the picornavirus family. They can
The two individuals most responsible for control of this terrifying be grown in vitro in cell cultures, where they cause cell destruc-
disease will not see its imminent elimination from the world— tion to form plaques. plaque assay, p. 317
Albert Sabin died in 1992; Jonas Salk in 1995. The two men
were bitter rivals, both of whom expected, but did not receive, the Pathogenesis
Nobel Prize. Polioviruses enter the body orally, infect the throat and intestinal
tract, and then invade the bloodstream. Usually, symptoms are
Signs and Symptoms mild or absent, the immune system conquers the infection, and
Poliomyelitis usually begins with symptoms of meningitis: head- recovery is complete. Only rarely does the virus bind to specific
ache, fever, stiff neck, and nausea. In addition, muscle pain and receptors on motor neurons, replicate, and destroy the cells when
spasm generally occur, followed by paralysis. Over the next the mature virus is released. Since most people infected with
weeks and months, muscles shrink and bones do not develop nor- poliovirus do not develop nervous involvement, a single case of
mally in the affected area. In severe cases, the respiratory muscles poliomyelitis means that the virus is rampant.
are paralyzed, and air must be pumped in and out of the lungs by
an artificial respirator. Those who survive this acute stage of the Epidemiology
illness recover some function. Sensory neurons are not affected. In areas where sanitation is poor, the polioviruses are endemic,
People who survive poliomyelitis sometimes develop post- transmitted by the fecal-oral route. Newborns in these nations
polio syndrome. This condition is characterized by muscle pain, are partially protected for 2 to 3 months until antibodies received
increased weakness, and muscle degeneration 15 to 50 years from their mothers begin to wane. During this time, the infants are
after recovering from acute poliomyelitis. Post-polio syndrome is usually exposed to poliovirus because of crowding and unsanitary
not due to a resurgence of polioviruses and sometimes involves conditions. They develop mild infections of the throat and intes-
muscles not obviously affected by the original illness. Instead it tine, but the maternal antibodies generally prevent the virus from
is thought to be a secondary effect of the initial damage. During spreading to the motor neurons. Thus, infants develop lifelong
recovery from acute poliomyelitis, surviving nerve cells branch immunity to the virus during the time when the maternal antibod-
out to take over the functions of the killed nerve cells. Post-polio ies protect them from paralysis.
 Part IV Infectious Diseases 657

Poliomyelitis has been most devastating in countries with

100 20

Cases reported
good sanitation. In these situations, polioviruses generally dis- Inactivated
virus (Salk) 15
appear from the community because they cannot spread to sus- vaccine 10
ceptible people. Then, when poliovirus is reintroduced, a high
10 5
incidence of paralysis results because people lack immunity. This

Cases per 100,000 population

occurred in the United States in the 1950s, resulting in many cases 0
1990 1995 2000 2005
of respiratory paralysis and death (figure 26.13). With most peo- Year
ple now routinely immunized against the disease, however, and 1
the likelihood of imported disease rapidly waning, this scenario
will hopefully no longer occur. Attenuated
0.1 oral (Sabin)
Treatment and Prevention vaccine

There is no treatment for polio, but individuals receive supportive

care such as rest, intravenous fluids, and pain medications. If the 0.01
respiratory muscles are paralyzed, use of a ventilator or “iron
lung” is required. Exercise therapy may be helpful to those with
post-polio syndrome. 0.001
Like other enteroviruses, polioviruses are quite stable under 1951 1955 1960 1965 1970 1975 1980 1985 1990
natural conditions, and can often be found in swimming pools. Year
They can be inactivated by pasteurization and by chlorinating
drinking water. Control of poliomyelitis using vaccines represents FIGURE 26.14 Incidence of Poliomyelitis in the United
States, 1951–2007 Ironically, since 1980, all cases acquired in
one of the greatest success stories in the battle against infectious the United States have been caused by the oral attenuated (Sabin)
diseases (figure  26.14). Ironically, all cases of paralytic polio vaccine. Endemic poliomyelitis was eliminated from the United States
acquired in the United States since 1980 were caused by Sabin’s by 1980, and the entire Western Hemisphere by 1991.
oral, attenuated polio vaccine, which was introduced in 1961. ? Why is the Sabin vaccine against poliomyelitis the preferred
These cases arise because in rare instances the vaccine strain vaccine to use in countries where polio is still prevalent?
mutates and becomes virulent.
The small risk of developing paralytic poliomyelitis from where transmission of the wild virus still occurs. It produces bet-
Sabin’s vaccine virus (approximately one case per 2.4 million ter mucosal immunity in the throat and intestine, can spread from
doses given) led the United States to return to the routine use of person to person, does not require an injection, provides herd
the inactivated Salk vaccine in 1999. However, the Sabin oral immunity, and is less expensive than the Salk vaccine. In the years
vaccine remains key to controlling polio in areas of the world following 1988, when the World Health Organization resolved to
eradicate poliomyelitis, the number of countries with endemic dis-
ease has been reduced from 125 to only four (Afghanistan, India,
Nigeria, and Pakistan). Poliomyelitis is summarized in table 26.9.
herd immunity, p. 421 campaign to eliminate poliomyelitis, p. 424

TABLE 26.9 Poliomyelitis

Signs and Headache, fever, stiff neck, nausea, pain,
symptoms muscle spasm, followed by paralysis
Incubation period 7 to 14 days
Causative agents Polioviruses 1, 2, 3, members of the
picornavirus family
Pathogenesis Virus infects the throat and intestine,
circulates via the bloodstream, and enters
some motor nerve cells of the brain or spinal
cord; infected nerve cells lyse upon release of
mature virus.
Epidemiology Spreads by the fecal-oral route; asymptomatic
and nonparalytic cases common

FIGURE 26.13 The Horror of Poliomyelitis These tanklike Treatment Treatment: artificial ventilation for
respirators (“iron lungs”) were used during the 1950s epidemics of and prevention respiratory paralysis; physical therapy and
poliomyelitis to keep alive people whose respiratory muscles were rehabilitation. Prevented by injecting Salk’s
paralyzed by the disease. Now we can hope to see polio forever inactivated vaccine or by Sabin’s orally
banished from the earth! administered attenuated vaccine in areas of
epidemic or endemic disease.
? How are polioviruses transmitted?
658 Chapter 26 Nervous System Infections

Rabies
Rabies is a classic zoonotic disease—one normally found in
animals—but it can be transmitted to humans. In the United
States, immunization of dogs and cats against rabies has
practically eliminated them as a source of human disease.
Still, rabies has multiple wild animal hosts. These remain
a constant threat to non-immunized domestic animals and
humans. The rabies virus typically enters the body through
a bite wound and attacks the nervous system. Rabies is
unusual in that it can be effectively prevented with a vac-
cine given shortly after exposure to the virus. Without
such treatment, however, death is almost certain. zoonotic
disease, p. 439

Signs and Symptoms

Rabies begins with fever, head and muscle aches, sore throat, 75 nm
fatigue, and nausea. The characteristic symptom is a tingling or (a)
twitching sensation at the site of viral entry, usually an animal
bite. These symptoms generally do not appear until 1 to 2 months
after infection, but they progress rapidly to agitation, confusion,
hallucinations, seizures, increased sensitivity, and encephalitis.
A  few days later, the individual typically goes into a coma and
dies of respiratory failure or cardiac arrest.
The later stages of rabies are often characterized by increased
salivation and difficulty in swallowing. This causes the “frothing
at the mouth” classically associated with rabid animals and “mad
dogs.” Swallowing, or even the sight of fluids, often leads to
severe spasms of the throat and respiratory muscles. The common
name for rabies is “hydrophobia” or fear of water.

Causative Agent
20 μm
The cause of rabies is the rabies virus (figure 26.15a), a member (b)
of the rhabdovirus family. This virus has a striking bullet shape, FIGURE 26.15 Rabies Virus (a) Color-enhanced transmission
is enveloped, and contains single-stranded, negative-sense RNA. electron micrograph of rabies virus. Notice the bullet shape.
(b) Stained smear of brain tissue from a rabid dog. A Negri body
Pathogenesis (see arrow) represents a site of rabies virus replication.
After the rabies virus is introduced into the body, it typically ? Why can an animal that appears well still transmit the
multiplies in cells at the site of infection for several weeks before rabies virus?
entering a sensory neuron. It then travels by retrograde transport
up the axon to the spinal cord and eventually to the brain. The Epidemiology
length of time before symptoms occur depends on the location of The primary mode of transmission of rabies to humans is via the
the bite, the amount of virus introduced, and the condition of the saliva of a rabid animal introduced into bite wounds of the skin.
host. Individuals with head wounds, for example, tend to show It has been reported, but not documented, that individuals can
symptoms sooner than those with leg wounds. also contract rabies by inhaling aerosols containing the virus,
Once in brain tissue, the virus multiplies extensively, caus- such as from bat feces. Because of the extensive rabies vaccina-
ing the symptoms of encephalitis. Characteristic inclusion bodies tion program for dogs in the United States, the main reservoir for
called Negri bodies, made up of viral nucleocapsids, are found rabies is wild animals such as raccoons, bats, skunks, and foxes
in most rabies cases (figure  26.15b). From the brain, the virus (figure 26.16). Over 5,000 wild animal cases are reported in
spreads outward via the nerves to various body tissues, notably the the United States each year, representing an enormous reservoir
salivary glands, eyes, and fatty tissue under the skin, as well as to from which infection can be transmitted to domestic animals and
the heart and other vital organs. Rabies can be diagnosed before humans. Raccoons lead the list of wildlife cases, but almost all
death by identifying the virus in stained smears collected from the human cases are due to contact with infected bats. Human rabies
surface of the eyes. is rare in the United States with only 1 to 3 cases reported per year.
Although rabies is rare in the United States, about 40,000-
MicroByte
The only documented cases of human-to-human transmission of rabies 70,000 people worldwide die of rabies. Most of these are due to
have been in patients receiving transplants from infected donors. dog bites in areas where dogs are not routinely vaccinated. A per-
son bitten by a dog with rabies virus in its saliva has about a 30%
 Part IV Infectious Diseases 659

PERSPECTIVE 26.1
Rabies Survivors!
In October 2004, a 15-year-old girl was admit- after she returned home. So far, attempts to Survival after the onset of rabies symp-
ted to the hospital with typical signs and cure other patients using the treatment used in toms is known to have occurred in only five
symptoms of rabies. A month earlier, while this case have been unsuccessful. individuals prior to these cases, and all five had
in church, she picked up a bat that had fallen In February 2009, a 17-year old girl reported received anti-rabies vaccine either before they
to the floor. The bat bit her left index finger, to the emergency room with a severe headache were exposed to rabies or before they showed
and she carried it outside and released it. The that had lasted for 2 weeks, vomiting, neck signs or symptoms of the disease. All but one
small bite wound was cleaned with hydrogen pain, and other symptoms of nervous system suffered persisting neurological damage, unlike
peroxide and healed uneventfully. In the hos- infection. On March 6, she was admitted to the these patients, who made complete recoveries.
pital, a tube was placed in the girl’s trachea hospital with suspected infectious encephalitis Over the years, many treatments have been
and hooked to a respirator because most rabies but did not respond to treatment. Two months tried without benefit, including human rabies
deaths result from respiratory failure. She was earlier she had entered a cave in Texas and immune globulin, anti-rabies vaccines, and
given medication to put her into a coma to rest several flying bats had hit her body, although interferon. What is to be learned from the sur-
her nervous system, and she was given the she did not notice any scratches or bite wounds vival of these patients? Perhaps their survival
antiviral medication ribavirin, according to an at the time. On March 11, she tested positive had nothing to do with their treatment—but
experimental protocol. After 7 days, the coma- for rabies and was given hyperimmune human was due to their own immune systems fighting
inducing medication was reduced and the girl rabies globulin and one dose of anti-rabies a small infectious dose or a rabies viral strain
was allowed to wake up. A little more than a vaccine. She remained in the hospital with of low virulence. Is there some clue in their sur-
month after her illness began, her breathing supportive care, but never entered intensive vival that will perhaps lead to the first effective
tube was removed, she regained speech, solved care and was discharged on March 22. After an rabies treatment in the 3,000 years the disease
mathematics problems, and walked with assis- emergency room visit on April 3 for headache has been recognized?
tance. Slow, steady improvement continued and vomiting, she did not return for follow-up.

risk of developing rabies. Most rabies cases are in Asia, followed simply stops eating, becomes inactive, and suffers paralysis of
by Africa. Rabies does not exist in Australia or New Zealand. throat and leg muscles. Obviously, one should not try to remove a
Most rabid dogs excrete the virus in their saliva, sometimes foreign body from the throat of a sick, choking, unvaccinated dog!
even a few days before they get sick. Therefore, if an unvaccinated
dog bites a person, the animal should be confined for 10 days to Treatment and Prevention
see if it develops symptoms of rabies. Some dogs become irritable A person who has been bitten by an animal should immediately
and hyperactive with the onset of rabies, produce excessive saliva, wash the wound thoroughly with soap and water, and then apply
and attack people, animals, and inanimate objects. Perhaps more an antiseptic to avoid any kind of infection. If there is a possibil-
common is the “dumb” form of rabies, in which an infected dog ity that the animal is rabid, human rabies immune globulin (anti-
rabies antibody) is injected at the wound site and intramuscularly
to provide passive immunity. The individual should then receive
FIGURE 26.16 four injections of inactivated vaccine, the first as soon as possible
Rabies Carrier?
Skunks are a main
after exposure, and at 3, 7, and 14 days after the first. The vaccine
reservoir for rabies. provokes an immune response that neutralizes free virus and kills
infected cells during the long incubation period before the virus
? What other
animals most enters the neurons. passive immunity, p. 420
commonly carry There is no effective treatment for rabies once symptoms
the rabies virus? appear. Only two people are known to have recovered from the
disease without receiving the vaccine (Perspective 26.1). Active
immunization after a bite can usually generate antibody in time to
protect against the disease. In the United States, about 30,000 peo-
ple annually receive inactivated rabies vaccine after a bite from a
suspected rabid animal. Individuals at high risk for rabies, such as
veterinarians or animal control personnel, should be immunized
before exposure.
It is impossible to eradicate a zoonotic disease without clear-
ing the animal hosts. Routine vaccination of domestic dogs and
cats in the United States has dramatically reduced the number of
cases in domestic pets. A DNA vaccine has been developed that
may make it more practical to vaccinate dogs in developing coun-
tries with high incidence of rabies. Vaccination of wild animals
is more difficult since it is impossible to catch, immunize, and
660 Chapter 26 Nervous System Infections

Inhalation of fungal cells found in soil and bird droppings seldom

TABLE 26.10 Rabies
causes serious lung disease, but phagocytic cells that contain these
Signs and Fever, headache, nausea, vomiting, sore cells may carry them from the lungs via the bloodstream to the
symptoms throat, cough at onset; later, spasms of the brain. This can lead to inflammation of the brain and meninges.
muscles of mouth and throat, coma, and
death
Incubation Usually 30 to 60 days; sometimes many Cryptococcal Meningoencephalitis
period months or years
Cryptococcal meningoencephalitis was uncommon until the
Causative agent Rabies virus, single-stranded RNA,
onset of the AIDS epidemic. Now the disease is among the most
rhabdovirus family; has an unusual bullet
shape important HIV-related opportunistic infections. Because it is dif-
ficult to clear without T-cell involvement, AIDS patients with
Pathogenesis During incubation period, virus multiplies
at site of bite, then travels via nerves to this infection are sometimes maintained indefinitely on antifungal
the central nervous system; it multiplies medications. More recently, the incidence of cryptococcal menin-
and spreads outward via multiple nerves to goencephalitis has increased among healthy individuals in the
infect heart and other organs. western United States and Canada due to an emerging pathogen,
Epidemiology Bite of rabid animal, usually a bat. Cryptococcus gattii. opportunistic infection, p. 382
Inhalation is another possible mode.
Treatment Effective post-exposure measures: Signs and Symptoms
and prevention immediately wash wound with soap and In apparently healthy people, symptoms of cryptococcal meningo-
water and apply antiseptic; inject rabies
vaccine and human rabies antiserum as
encephalitis develop gradually and generally consist of difficulty
soon as possible. No effective treatment in thinking, dizziness, intermittent headache, and possibly slight
once symptoms begin. Avoid suspect fever. After weeks or months, vomiting, weight loss, paralysis,
animals; immunize pets. seizures, and coma may appear. In people with immunodefi-
ciency, the disease generally progresses much faster; without
treatment, death can occur in as little as 2 weeks.
release all of them. Programs are underway to administer an oral
vaccine to widespread wild populations by placing it into a bait Causative Agent
food, such as peanut butter or fish meal. Many eastern states have Cryptococcal meningoencephalitis is an infection of the meninges
participated in air-drop programs to vaccinate raccoons; Texas has and brain by either Cryptococcus neoformans or Cryptococcus
a similar program targeting foxes and coyotes. Some features of gattii. The organisms are small, spherical yeasts generally 3 to
rabies are summarized in table 26.10. 20 μm in diameter surrounded by a thick capsule that resists the
immune response (figure  26.17). Cryptococcus neoformans is
MicroAssessment 26.3
an opportunistic pathogen that usually causes disease only in the
Many different kinds of viruses can attack the nervous system, but immunocompromised. C. gattii, on the other hand, causes disease
they generally do so in only a small percentage of infected people. in healthy individuals.
At least half of viral meningitis cases are caused by the enterovirus
subgroup of picornaviruses, which are generally spread by the fecal-
oral route. Viral meningitis is usually mild, but viral encephalitis
often causes permanent disability. Arboviruses maintained in nature
in a mosquito-bird or mosquito-rodent cycle are a leading cause of
epidemic viral encephalitis. Sporadic viral encephalitis is usually due
to herpes simplex virus. Poliomyelitis, characterized by paralysis of
one or more muscle groups, is caused by three other enteroviruses.
Rabies is a widespread zoonosis, almost uniformly fatal for humans,
usually transmitted by animal bites.
7. Explain why epidemic viral encephalitis is called a zoonosis.
8. What are the advantages and disadvantages of the Sabin polio
vaccine?
9. Why is rabies now rare in humans when it is still so common
in wildlife? +

26.4 ■ Fungal Diseases

of the Nervous System
20 μm
Learning Outcome
FIGURE 26.17 Cryptococcus neoformans Note the capsule
10. Compare and contrast the development of cryptococcal around the organism.
meningoencephalitis in AIDS patients and in healthy individuals.
? How might a capsule benefit a yeast cell?
 Part IV Infectious Diseases 661

Pathogenesis TABLE 26.11 Cryptococcal Meningoencephalitis

Infection is first established in the lung, usually producing mild or
no symptoms. Immune defenses of healthy people usually elimi- Signs and Headache, vomiting, confusion, and weight
symptoms loss; slight or no fever; symptoms may
nate the infection, but phagocytic killing is slow and inefficient. progress to seizures, paralysis, coma, and
In some cases, particularly in immunocompromised individuals, death
the organisms multiply, enter the bloodstream, and are distributed Incubation period Widely variable, few to many weeks
throughout the body. Meningoencephalitis is the most common
Causative agent Cryptococcus neoformans, Cryptococcus
infection outside of the lung, but organisms in the bloodstream
gattii—encapsulated yeasts
can also infect skin, bones, and other body tissues. The capsule
Pathogenesis Infection starts in lung; encapsulated
inhibits the antibody response and phagocytosis of the organisms.
organisms multiply, enter bloodstream, and
Capsular material can be detected in spinal fluid and urine, aiding are carried to various parts of the body;
diagnosis. In meningoencephalitis, the organisms typically cause phagocytosis inhibited; meninges and
thickening of the meninges, sometimes hindering the flow of cere- adjacent brain tissue become infected.
brospinal fluid and increasing pressure within the brain. They also Epidemiology Inhalation of material contaminated with
invade brain tissue, producing multiple abscesses. the fungus; other sources; most people
resistant to the disease
Epidemiology Treatment Treatment: amphotericin B with flucytosine
and prevention followed by fluconazole. No preventive
Cryptococcus neoformans is distributed worldwide in soil and
measures.
vegetation contaminated with bird droppings. Symptomatic infec-
tion is sometimes the first indication of AIDS. Cryptococcus
gattii was once associated with eucalyptus trees in tropical or toxic effects. Because amphotericin B does not reliably cross
subtropical regions of the world, but emerged in 1999 in British the blood-brain barrier, the drug may be administered through a
Columbia, Canada. Pathogenic strains have since spread in the plastic tube inserted through the skull into a lateral ventricle of
U.S. Pacific Northwest and Canada (figure 26.18). Appearance of the brain. Treatment is successful in about 70% of cases except
more virulent strains has increased the case-fatality rate from 5% in AIDS patients who respond poorly to treatment, most likely
to about 25%. because they lack T-cell-dependent killing that normally assists
The organisms enter the body by inhalation of spores; infec- the action of the antifungal medications. Unless their T-cell func-
tion can occur in humans as well as cats, dogs, and other animals. tion can be restored by treatment, AIDS patients are rarely cured
Subsequent onset of disease is rare. Person-to-person transmission of their infection. antifungal medicines, p. 476
of the disease does not occur. There is no vaccine or other preventive measure available.
The main features of cryptococcal meningoencephalitis are sum-
MicroByte marized in table 26.11.
Several Dall’s porpoises have been found washed up on the shores in
British Columbia since 2000; all were positive for C. gattii infection.
MicroAssessment 26.4
Cryptococcal meningoencephalitis occurs opportunistically in
Treatment and Prevention immunocompromised individuals such as AIDS patients. The
Treatment with the antibiotic amphotericin B is often effective, causative organism is a small yeast with a large capsule, often found
particularly if given with flucytosine (5-fluorocytosine) followed in soil contaminated with pigeon droppings. The disease has recently
emerged among healthy individuals in the Pacific Northwest.
by the oral medicine, fluconazole. Amphotericin B must be given
intravenously and the dose carefully regulated to minimize its 10. Why is it difficult to cure AIDS patients of a fungal infection?
11. Why are AIDS patients so vulnerable to cryptococcal
meningoencephalitis?
FIGURE 26.18 12. What might cause the spread of new strains of C. gattii in the
British Columbia Spread of northwest United States? +
Mainland Cryptococcus
gattii in
Vancouver the Pacific
Island
Northwest 26.5 ■ Protozoan Diseases
in 2008
of the Nervous System
Washington ? How does
Cryptococcus Learning Outcome
gattii enter
the body? 11. Compare and contrast African sleeping sickness and primary
amebic meningoencephalitis.

Infection of the nervous system by protozoans is quite rare, but

Oregon usually results in fatal disease when it occurs. Protozoan diseases
are difficult to treat because medications that affect eukaryotic
cells may also affect fragile neurons. protozoa, p. 291
662 Chapter 26 Nervous System Infections

African Sleeping Sickness Pathogenesis

African sleeping sickness—also known as African The protozoan enters the bite wound in the saliva of an infected
trypanosomiasis—is transmitted by the tsetse fly, its biological tsetse fly. The parasite multiplies at the skin site and eventually
vector. Sickness may persist for years, producing the symptoms enters the lymphatics and blood circulation. Immune responses
for which it is named. The disease is important because it can of fever and antibody production lessen early symptoms. Within
be contracted by residents and visitors in a wide area across the about a week and at roughly weekly intervals thereafter, however,
middle of the African continent. biological vector, p. 442 the number of parasites in the blood increases. Each burst coin-
cides with the appearance of a new glycoprotein on the surface of
Signs and Symptoms the trypanosomes. The parasite is a master at avoiding the immune
response, with more than a thousand different genes coding for
African trypanosomiasis can be chronic or acute. A tender nodule
variations of that surface glycoprotein, but only one is expressed
develops at the site of the bite within a week after a person is bit-
at a time. Each time a new gene is expressed, a new immune
ten by an infected tsetse fly. Regional lymph nodes might enlarge,
response is needed to produce the appropriate antibody. The recur-
but symptoms can disappear spontaneously. In the chronic form of
rent cycles of parasitemia and antibody production continue until
the disease, recurrent fevers develop that can continue for months
the patient is treated or dies.
or years. Involvement of the central nervous system is marked
In T. brucei rhodesiense infections, disease can progress
by gradual loss of interest in everything, decreased activity,
rapidly. The parasite enters the heart and brain within 6 weeks
and indifference to food. The eyelids droop, the individual falls
of infection. Irritability, personality changes, and mental dullness
asleep even while eating or standing, and speech becomes slurred.
result from brain involvement, but the patient usually dies from
Eventually the person becomes comatose and dies. In acute cases,
heart failure within 6 months. With T. brucei gambiense, progres-
symptoms develop over weeks or months, and neurological symp-
sion of the chronic infection is much slower. Years may pass
toms develop much more rapidly.
before death occurs, often from secondary infection. Much of the
damage to the host is due to immune complexes formed when high
Causative Agent
levels of protozoan antigen are bound by antibodies that then react
African sleeping sickness is caused by the flagellated protozoan, with complement system proteins. immune complex, p. 395
Trypanosoma brucei. These organisms are slender and have a
wavy, undulating membrane and an anteriorly protruding fla- Epidemiology
gellum (figure  26.19). Two subspecies are morphologically
African sleeping sickness occurs on the African continent within
identical—T. brucei rhodesiense and T. brucei gambiense. The
about 15° of the equator, with 10,000 to 20,000 new cases each
rhodesiense subspecies causes acute disease and occurs mainly in
year, some in tourists. The acute Rhodesian form of the disease
the cattle-raising areas of East Africa; the gambiense subspecies
is a zoonosis, and the main reservoirs are wild animals. Humans
causes chronic disease and occurs mainly in forested areas of
are the main reservoir for the chronic Gambian form, and human-
Central and West Africa. Both are transmitted by tsetse flies, bit-
to-human transmission is more common. The presence of animal
ing insects of the genus Glossina.
hosts makes the Rhodesian form more difficult to control. Less
than 5% of the tsetse fly vectors are infected. reservoir, p. 438

Treatment and Prevention

As with other eukaryotic pathogens, treatment is problem-
atic because of toxic side effects of the available medications.
Nevertheless, treatment of infected people helps reduce the proto-
zoan’s reservoir. A single, intramuscular injection of the medica-
tion pentamidine prevents the Gambian form of the illness for a
number of months, although the infection could progress later.
Suramin can be used if the disease has not progressed to involve
the central nervous system; melarsoprol and eflornithine cross the
blood-brain barrier and can be used when the central nervous sys-
tem is involved. Early diagnosis and treatment of the Rhodesian
form is important to prevent rapid damage to the nervous system.
antiprotozoan medications, p. 478
Actions directed against tsetse fly vectors include use of
Trypanosomes Leukocyte 25 μm
insect repellents and protective clothing to prevent bites, use of
FIGURE 26.19 Trypanosoma brucei in a Blood Smear Notice traps containing bait and insecticides to reduce the vector popu-
the slender protozoa among the blood cells of an individual with lation, and clearing of brush to reduce breeding habitats for the
African sleeping sickness (African trypanosomiasis). flies. The main characteristics of African sleeping sickness are
? Name the biological vector of Trypanosoma brucei. presented in table 26.12.
 Part IV Infectious Diseases 663

TABLE 26.12 African Sleeping Sickness

Signs and Tender nodule at site of tsetse fly bite;
symptoms fever, enlargement of lymph nodes; later,
involvement of the central nervous system,
uncontrollable sleepiness, headache, poor
concentration, unsteadiness, coma, death
Incubation period Weeks to several years
Causative agent Trypanosoma brucei, a flagellated protozoan
Pathogenesis The protozoa multiply at site of a tsetse
fly bite, then enter blood and lymphatic
circulation; as new cycles of parasites are
released, their surface protein changes and
the body is required to respond with a new
antibody.
Epidemiology Bites of infected tsetse flies transmit the
trypanosomes through fly saliva; wild animal FIGURE 26.20 Naegleria fowleri Notice the suckerlike hooks
reservoir for T. brucei rhodesiense. used in phagocytosis. Keep Your Head Above Water sign: Courtesy of Jay
Stahl-Herz, MD
Treatment Treatment: suramin; when central
and prevention nervous system is involved, melarsoprol or ? By what route does Naegleria fowleri enter the brain?
eflornithine. Protective clothing, insecticides,
clearing of brush where flies breed,
pentamidine. Epidemiology
Naegleria fowleri may be carried in the normal microbiota, but
it rarely causes disease. For every case of Naegleria meningoen-
cephalitis, many millions of people are exposed to the organism
without harm. PAM is usually acquired when individuals swim
Primary Amebic or dive in warm natural fresh water. It is not found in seawater,
Meningoencephalitis (PAM) and chlorination in swimming pools will kill the organism.
Contaminated drinking water cannot spread the pathogen, and it
Naegleria fowleri, the cause of primary amebic meningoencepha- is not transmitted from person to person.
litis (PAM), is commonly found in warm fresh water and soils.
Nevertheless, fewer than 200 cases of PAM have been reported MicroByte
worldwide. About three people per year become infected after A person is over 10,000 times more likely to die from drowning in
swimming or diving in natural waters in the United States, making natural waters than from PAM.
it a rare event, but one that is almost certainly fatal.
Treatment and Prevention
Signs and Symptoms
The antifungal drug amphotericin B has been used in these infec-
Symptoms of PAM are similar to those of bacterial meningitis. tions, but with little success. There is no vaccine. The main char-
Early symptoms include headache, fever, stiff neck, and vomit- acteristics of primary amebic meningoencephalitis are presented
ing. Once neurological symptoms appear—such as confusion or in table 26.13.
seizures—death quickly follows.

Causative Agent
Primary Amebic
Naegleria fowleri is one of only a few free-living protozoa TABLE 26.13 Meningoencephalitis (PAM)
pathogenic for humans. This frightening-looking organism with
sucker-like structures used in phagocytosis can literally “eat your Signs and symptoms Headache, fever, stiff neck, and
brain” (figure 26.20). The ameboid trophozoite gives rise to vomiting, confusion, seizures
flagellated forms and spherical cysts (see figure 12.14). It prefers Incubation period Less than a week
warm temperatures. Causative agent Naegleria fowleri
Pathogenesis Destroys brain tissue
Pathogenesis Epidemiology Rare incidence after swimming or
Naegleria fowleri penetrates the skull along the olfactory nerves diving in warm, natural waters
serving the nasal mucosa. It multiplies and migrates to the brain, Treatment Unsuccessful treatment with
where it destroys nervous tissue, especially in the frontal lobes. and prevention amphotericin B; no vaccine
Hemorrhage, coma, and death occur within a week.
664 Chapter 26 Nervous System Infections

MicroAssessment 26.5 spongiform encephalopathy to humans who eat them. chronic

wasting disease, p. 328
Residents and visitors to a wide swath of tropical Africa are at risk
of contracting African sleeping sickness, caused by the flagellated
protozoan parasite Trypanosoma brucei, and transmitted by a Transmissible Spongiform
biting insect, the tsetse fly. These protozoa can circulate in the Encephalopathy in Humans
bloodstream for extended periods by changing their surface antigens
to escape the host’s antibodies. Eventually they penetrate the CNS, Transmissible spongiform encephalopathy (TSE) is rare in
causing indifference, sleepiness, coma, and death. Only on rare humans, occurring in only 0.5 to 1 case per million people.
occasions can free-living amebas such as Naegleria fowleri cause Most cases occur as Creutzfeldt-Jakob disease (CJD), which
meningoencephalitis. affects individuals over 45 years of age and sometimes runs
13. How likely is it that a person who swims in warm fresh water in families. The disease acquired by eating affected animals is
will contract primary amebic meningoencephalitis? distinctively different—but has the same result and is identified
14. How can one explain repeated, abrupt increases in T. brucei in as a variant of CJD, termed vCJD. Cases of CJD and vCJD are
the blood of African sleeping sickness victims? + invariably fatal.

MicroByte
26.6 ■ Diseases Caused by Prions Another human TSE, kuru, is associated with cannibalism in New
Guinea, where natives ate deceased relatives as a sign of respect.
Learning Outcomes
12. Explain how prions differ from other infectious agents. Signs and Symptoms
13. Describe how prions can cause disease. Early symptoms of TSE include vague behavioral changes, anxi-
ety, insomnia, and fatigue. Symptoms progress to characteristic
A rare and mysterious group of chronic, degenerative brain muscle jerks, lack of coordination, memory loss, and dementia.
diseases caused by prions has been seen in wild animals (mink, Although the incubation period may last for years, once symptoms
elk, and deer), domestic animals (sheep, goats, and cattle), and appear, death generally occurs within a year.
humans. Affected brain tissue has a spongy appearance, which is
why these diseases are referred to as spongiform encephalopathies Causative Agent
(figure 26.21). The causative agents of spongiform encephalopathies are called
Scrapie, a disease of sheep and goats, was named because proteinaceous infectious particles, or prions. Much smaller than a
affected animals had difficulty standing and “scraped” along virus, prions (PrP) appear to be a misfolded form of a normal cel-
fences for support. Cattle, presumably fed meat and bone meal lular protein (PrPc). PrP is encoded by a normal human gene and
from infected sheep, developed bovine spongiform encepha- modified after transcription. The misfolding results in a protein
lopathy, or “mad cow disease.” These diseases assumed new that is protease-resistant, whereas the normal protein is protease-
prominence in the 1990s when an outbreak of a spongiform sensitive. prions, p. 328
encephalopathy in humans in the United Kingdom was related
to an earlier outbreak of mad cow disease. Brain and other tis- Pathogenesis
sues from affected animals can transmit the disease to normal
Prions increase in quantity during the incubation period of the
animals, even of different species. Although it has not been
disease as the misfolded protein (PrP) acts as a template that
documented, there is some concern that wild animals with
promotes misfolding of the normal cellular protein (PrPc) on
chronic wasting disease caused by prions may also transmit
the surface of neurons (see figure 13.27). Prions aggregate in
insoluble masses called plaques in the brain, causing tissue
damage. They may be taken up by neurons or phagocytic cells,
but cannot be degraded by cellular proteases. Death of neurons
produces the spongy appearance of affected brain tissue. Not
all prions, however, act the same. They differ in host range,
incubation period, and the areas of the nervous system attacked.
Transmissible spongiform encephalopathies can be differenti-
ated from encephalitis because they typically do not evoke an
immune response. This may be because the plaques consist of
host protein.

Epidemiology
Normal Infected Creutzfeldt-Jakob disease (CJD) generally occurs in individuals
FIGURE 26.21 Appearance of Brain with Spongiform older than 45 years. It has been transmitted from human to human
Encephalopathy Notice the spongelike appearance of the infected through corneal transplants, contaminated surgical instruments,
brain on the right compared to the normal brain on the left. and injections of human hormone replacements. It can be trans-
? What animals are affected by spongiform encephalopathies? mitted experimentally to chimpanzees.
 Part IV Infectious Diseases 665

Transmissible Spongiform individuals with vCJD is only 28 years. When mad cow disease
TABLE 26.14 Encephalopathy is suspected, radical measures may minimize its transmission to
humans. In 2005, for example, the United States banned impor-
Signs and Behavioral changes, anxiety, insomnia, tation of cattle from Canada when the disease was identified in
symptoms fatigue, progressing over weeks or months to several animals from there.
muscle jerks, lack of coordination, dementia
Incubation period Usually many years Treatment and Prevention
Causative agents Proteinaceous infectious particles known as There is no treatment for the spongiform encephalopathies, and
prions; lack nucleic acids; identical amino they are always fatal. It is important to avoid eating any animals
acid sequence to a normal protein, but
that show neurological symptoms. Prions are highly resistant to
folded differently, and relatively resistant to
proteases; resistant to heat, radiation, and disinfectants, including formaldehyde. They also are resistant to
disinfectants heat and to ultraviolet and ionizing radiation. They can be inacti-
Pathogenesis Prions increase in quantity by converting vated by extended autoclaving in 1M (molar) sodium hydroxide.
normal protein to more prions; transmission The main features of transmissible spongiform encephalopathies
to the brain; aggregation into masses outside are presented in table 26.14.
the nerve cells; cell malfunction and death.
Epidemiology Human-to-human transmission by corneal The key features of the diseases covered in this chapter are high-
transplantation and by contaminated surgical lighted in the Diseases in Review 26.1 table.
instruments; probable transmission of cattle
prions to humans by eating contaminated
beef; sporadic Creutzfeldt-Jakob disease MicroAssessment 26.6
in those over age 45 years; median age of Transmissible spongiform encephalopathies are degenerative nervous
variant Creutzfeldt-Jakob cases only 28 years.
system diseases that occur in a variety of wild and domestic animals.
Treatment No treatment, invariably fatal. Prions are They are rare in humans, but disease can be transmitted from animals
and prevention inactivated by autoclaving in concentrated via ingested tissues. These diseases appear to be caused by prions—
sodium hydroxide. infectious agents consisting only of protein and highly resistant to
inactivation by heat, radiation, and disinfectants. The diseases are
always fatal.
Spongiform encephalopathy of sheep (scrapie) has been 15. What is a prion?
known for more than two centuries, without any evidence that it
16. What is the best way to prevent transmissible spongiform
is directly transmissible to humans. Current evidence indicates encephalopathies?
that cattle prions can be transmitted to humans and cause a vari-
17. If you were an eye surgeon, would you rather the donor for
ant Creutzfeldt-Jakob disease (vCJD) marked by differences in a cornea transplant be under 35 or over 45 years of age? +
symptoms, brain pathology, and age of onset. The median age of

Diseases in Review 26.1

Nervous System Diseases

Summary
Disease Causative Agent Comment Table
BACTERIAL NERVOUS SYSTEM DISEASES

Pneumococcal meningitis Streptococcus Leading cause of meningitis in adults; 90 serotypes with vaccine to Table 26.2,
pneumoniae 23 of the most common. p. 649
(pneumococcus)
Meningococcal Neisseria May lead to endotoxic shock; formation of petechiae due to capillary Table 26.1,
meningitis meningitidis damage; associated with meningitis epidemics; vaccine against the p. 646
(meningococcus) common serotypes is available, but occurs in unvaccinated children
and young adults.
Haemophilus influenzae Haemophilus Once the most common cause of infant meningitis, now largely Table 26.2,
meningitis influenzae controlled by a conjugate vaccine. p. 649

(continued)
Diseases in Review 26.1 (Continued )
Nervous System Diseases

Summary
Disease Causative Agent Comment Table
BACTERIAL NERVOUS SYSTEM DISEASES (Continued)
Neonatal meningitis Streptococcus Newborns can acquire causative agent from the mother’s birth canal; Table 26.2,
agalactiae U.S. pregnant women are routinely screened for the organism before p. 649
delivery and if positive treated with antibiotics.
Listeriosis Listeria Contaminated foods have caused epidemics; multiplies at Table 26.3,
monocytogenes refrigeration temperatures; infected pregnant women may miscarry; p. 650
may cause abscesses in the fetus.
Hansen’s disease Mycobacterium Infects peripheral nerves, causing immune system to attack them; Table 26.4,
(leprosy) leprae course of disease determined by cell-mediated immune response; p. 652
long incubation period; loss of limbs, blindness.
Botulism Clostridium Botulinum toxin causes flaccid paralysis; foodborne intoxication is Table 26.5,
botulinum most common form worldwide; also occurs when organism grows in p. 653
wounds or in intestines of infants.

VIRAL NERVOUS SYSTEM DISEASES

Viral meningitis Usually enteroviruses Aseptic meningitis; more common and much milder than bacterial Table 26.6,
meningitis; often transmitted by fecal-oral route. p. 654
Viral encephalitis Arboviruses Transmitted in the United States by mosquitoes; more likely to cause Table 26.8,
death or disability than viral meningitis; case-fatality rate varies from p. 656
2–50%, depending upon type of virus.
Poliomyelitis Polioviruses Destruction of motor neurons leads to paralysis; fecal-oral Table 26.9,
transmission, but infection rarely leads to disease; vaccination has p. 657
eliminated the disease in most parts of the world, and the disease is
now targeted for eradication.
Rabies Rabies virus Zoonotic disease; virus travels from bite wound through sensory Table 26.10,
neurons to CNS, causing a generally fatal infection; vaccination p. 660
shortly after exposure prevents disease.

FUNGAL NERVOUS SYSTEM DISEASES

Cryptococcal Cryptococcus Caused by inhalation of fungal cells carried by phagocytic cells to Table 26.11,
meningoencephalitis neoformans and the brain; can be an early sign of AIDS, but now also seen in healthy p. 661
C. gattii individuals where C. gattii is found.

PROTOZOAN NERVOUS SYSTEM DISEASES

African sleeping sickness Trypanosoma brucei Chronic (Gambian) or acute (Rhodesian); two subspecies of protozoa Table 26.12,
transmitted by tsetse fly; loss of interest, drooping eyes, coma, death; p. 663
early treatment is key in acute form.
Primary amebic Naegleria fowleri Swimming or diving in water containing the organism transmits this Table 26.13,
meningoencephalitis rare but deadly disease; travels to brain via olfactory neurons where it p. 663
(PAM) damages tissue; no vaccine or effective treatment.

PRION DISEASES

Transmissible Prions Caused by an accumulation of destruction-resistant misfolded Table 26.14,

spongiform proteins, resulting in death of neurons; may come from contaminated p. 665
encephalopathy animal tissues; no treatment; always fatal.
 Part IV Infectious Diseases 667

FUTURE CHALLENGES 26.1

Eradicate Polio: Then What?
It is hoped that poliomyelitis will be among to immunize individuals missed earlier. The vac- viruses for months or years). Also laboratory
the next ancient scourges to follow smallpox cine viruses then disappear from the population freezers around the world hold stocks of the
down the road to eradication. However, for a within a few months. Although a small percent- viruses, as well as fecal specimens that could
variety of political and scientific reasons, the age of the population suffers paralytic illness harbor them. It is even possible to synthesize
causative virus of smallpox still exists many from the vaccine, the population is protected a poliovirus in the laboratory. Lastly, vaccine
years after the last naturally acquired case of from virulent polioviruses until new generations viruses can change genetically and acquire full
the disease. Will it be any easier to rid the are born. If there were no polioviruses left in the virulence, as occurred in the Hispaniola polio
world of the polioviruses after the last case of world, there would be no need to immunize the epidemic of 2000–2001.
poliomyelitis occurs? new generations. The challenge is to have a continuous,
Dramatic progress toward polio eradication Potential sources of virulent polioviruses reliable, global polio surveillance system,
has mainly been accomplished using “immu- include wild viruses circulating in remote and maintain immunizations and strategically
nization days,” when the entire population in a populations, individuals with mild or atypi- located stockpiles of vaccine during what is
given area is immunized using attenuated (Sabin) cal illness, and long-term carriers (especially likely to be a long time after the last case of
vaccine, often with one or more follow-up days those with immunodeficiency, who excrete the paralytic polio occurs.

Summary
26.1 ■ Anatomy, Physiology, and Ecology (figure 26.1) Listeriosis (tables 26.2, 26.3)
The brain and spinal cord make up the central nervous system Listeriosis is caused by Listeria monocytogenes, a non-spore-forming,
(CNS); the peripheral nervous system (PNS) is composed of Gram-positive rod usually associated with foodborne illness. The
nerves and ganglia. Cerebrospinal fluid (CSF) is produced in bacterium is widespread, commonly contaminates foods such as non-
ventricles in the brain and flows out over the brain and spinal cord pasteurized milk, cold cuts, and soft cheeses, and can grow in refriger-
(figure  26.2). Meninges are the membranes that cover the brain and ated foods (figure 26.7). The bacteria readily penetrate the gastrointesti-
spinal cord. Infectious agents most often reach the CNS through the nal mucus membranes, enter the bloodstream, and infect the meninges.
bloodstream when they penetrate the blood-brain barrier.
Hansen’s Disease (Leprosy) (figure 26.8; table 26.4)
Hansen’s disease is characterized by invasion of peripheral nerves
26.2 ■ Bacterial Diseases of the Nervous System
by the acid-fast rod Mycobacterium leprae, which has not been cul-
Bacteria most often infect the membranes surrounding the brain, caus-
tivated in vitro (figure  26.9). The disease occurs in two main forms—
ing meningitis. Bacterial meningitis is uncommon; immunization has
tuberculoid and lepromatous, depending on the immune status of the
greatly decreased the incidence among children. In most cases, the caus-
individual.
ative bacterium is part of the normal respiratory microbiota.
Botulism (table 26.5)
Pneumococcal Meningitis (table 26.2)
Botulism is not a nervous system infection, but an intoxication, usu-
Streptococcus pneumoniae, or pneumococcus, is the most common
ally foodborne, that causes severe generalized paralysis. The causative
cause of meningitis in adults (figure 26.3). Symptoms are similar to other
bacterium, Clostridium botulinum, is an anaerobic, Gram-positive rod
forms of meningitis: cold symptoms followed by abrupt onset of fever,
that forms heat-resistant endospores (figure  26.10). Endospores that
severe headache, pain and stiffness of the neck and back, nausea, and
survive canning or other heat treatment of foods germinate, and the
vomiting.
bacteria multiply, releasing a powerful toxin into the food. Because
Meningococcal Meningitis (tables 26.1, 26.2) of strict controls on food processing, intestinal or infant botulism
Meningococcal meningitis caused by Neisseria meningitidis is asso- is now the most common form of the disease in the United States
ciated with meningitis epidemics. Small hemorrhages in the skin (figure  26.11). Wound botulism, caused when C. botulinum colonizes
(figure  26.4), deafness, and coma can occur. Shock results from the dirty wounds containing dead tissue, is rare.
release of endotoxin into the bloodstream.
26.3 ■ Viral Diseases of the Nervous System
Haemophilus influenzae Meningitis (table 26.2) Most viral nervous system infections are caused by human entero-
Haemophilus influenzae, once the leading cause of childhood bacte- viruses or by the viruses of certain zoonoses. Many common viruses
rial meningitis, is largely sporadic and mostly controlled by a vaccine of humans can occasionally infect the nervous system, including those
(figure 26.6). that cause infectious mononucleosis, mumps, measles, chickenpox, and
herpes simplex (“cold sores,” genital herpes).
Neonatal Meningitis (table 26.2)
Newborns most often acquire meningitis-causing bacteria from the Viral Meningitis (table 26.6)
mother’s genital tract shortly before or during birth. Infants who survive Viral meningitis is much more common than bacterial meningitis. It is
often face long-lasting consequences of their infection. generally a mild disease for which there is no specific treatment.
668 Chapter 26 Nervous System Infections

Viral Encephalitis (table 26.8) 26.5 ■ Protozoan Diseases of the Nervous System
Viral encephalitis has a high fatality rate and often leaves survivors Only a few protozoa infect the human nervous system.
with permanent disabilities. Herpes simplex virus is the most important African Sleeping Sickness (table 26.12)
cause of sporadic encephalitis; epidemic encephalitis is usually caused
African sleeping sickness is a major health problem in a wide area
by arboviruses (figure 26.12; table 26.7).
across equatorial Africa. In its late stages, it is marked by indif-
Poliomyelitis (figures 26.13, 26.14; table 26.9) ference, sleepiness, coma, and death. The disease is caused by
Destruction of motor nerve cells of the brain and spinal cord leads to Trypanosoma brucei (figure  26.19), a flagellated protozoan transmitted
paralysis, muscle wasting, and failure of normal bone development. by its biological vector, the tsetse fly. During infection, the organism
Post-polio syndrome occurs years after poliomyelitis, and it is probably shows bursts of growth, each appearing with different surface proteins.
caused by the death of nerve cells that had taken over for those killed Primary Amebic Meningoencephalitis (PAM) (table 26.13)
by the poliomyelitis virus.
PAM is rare but fatal. Infection with Naegleria fowleri (figure 26.20) can
Rabies (figure 26.15; table 26.10) occur after swimming in warm, fresh water.
Rabies is a widespread zoonosis transmitted to humans mainly through
the bite of an infected animal (figure 26.16). Once symptoms appear in 26.6 ■ Diseases Caused by Prions
an infected person, the disease is almost always fatal. Because of the Prions—abnormal proteins that are resistant to heat, radiation, and
long incubation period, prompt immunization with inactivated vaccine disinfectants—cause the spongiform encephalopathies, rare diseases
after a rabid animal bite is effective in preventing the disease. Passive characterized by a spongelike appearance of brain tissue caused by loss
immunization given at the same time increases protection. of nerve cells (figure 26.21). They afflict a variety of wild and domestic
animals as well as humans.
26.4 ■ Fungal Diseases of the Nervous System Transmissible Spongiform Encephalopathies
Fungi are usually opportunistic but can cause disease in healthy people. in Humans  (table 26.14)
Cryptococcal Meningoencephalitis (table 26.11) Examples of transmissible spongiform encephalopathies (TSEs)
Infection begins in the lung after a person inhales spores of include “mad cow disease,” Creutzfeldt-Jakob disease (CJD), and
Cryptococcus neoformans or Cryptococcus gattii, encapsulated yeasts variant Creutzfeldt-Jakob disease (vCJD), which can be transmitted to
that resist phagocytosis (figure 26.17). C. neoformans infects immunocom- humans from infected meat. There is no treatment for these diseases,
promised individuals, but C. gattii can cause disease in healthy people and they are invariably fatal.
(figure 26.18). Treatment of these diseases is usually difficult.

Review Questions
Short Answer Multiple Choice
1. What sign would differentiate meningococcal meningitis from 1. Which is the best way to prevent meningococcal meningitis in
pneumococcal meningitis? individuals intimately exposed to the disease?
2. Name and describe the organism that is the leading cause of bacte- a) Vaccinate them against Neisseria meningitidis.
rial meningitis in adults. b) Treat them with the antibiotic rifampin.
3. What measures can be undertaken to prevent neonatal meningitis? c) Culture their throat and hospitalize them for observation.
4. Why is listeriosis so important to pregnant women even though it d) Withdraw a sample of spinal fluid and begin antibacterial
usually causes them few symptoms? treatment if the cell count is high and the glucose level is low.
5. Can botulism be spread from person to person? e) Have them return to their usual activities, but seek medical
6. Give two ways in which viral meningitis usually differs from bac- evaluation if symptoms of meningitis occur.
terial meningitis. 2. Which of these statements concerning the causative agent of liste-
7. What is the difference between sporadic encephalitis and epidemic riosis is false?
encephalitis? Name one cause of each. a) It can cause meningitis during the first month of life.
8. Explain why the biggest impact of poliomyelitis in the 1950s b) It is a Gram-positive rod that can grow in refrigerated food.
occurred in countries with good sanitation. c) It is usually transmitted by the respiratory route.
9. Why is it possible to prevent rabies with vaccine given after expo- d) Infection commonly results in bacteremia.
sure? e) It is widespread in natural waters and vegetation.
10. If you contract African sleeping sickness on a visit to central
Africa, what type do you most likely have?
 Part IV Infectious Diseases 669

3. Which of these statements concerning Hansen’s disease is false? 8. Which of these statements concerning cryptococcal meningoen-
a) It was once common in the United States. cephalitis is true?
b) An early symptom is loss of sensation, sweating, and hair in a a) It is caused by a yeast with a large capsule.
localized patch of skin. b) It is a disease of trees transmissible to humans.
c) The incubation period is usually less than 1 month. c) Typically it attacks the meninges but spares the brain.
d) Treatment should include more than one antimicrobial d) Person-to-person transmission commonly occurs.
medication given at the same time. e) It is seen only in persons who are immunocompromised.
e) The form the disease takes depends on the individual’s immune 9. Which of these statements concerning African sleeping sickness is
status. true?
4. Which of these statements concerning foodborne botulism is a) It is transmitted by a species of biting mosquito.
false? b) It is a threat to visitors to tropical Africa.
a) It is not a central nervous system infection. c) The onset of sleepiness is usually within 2 weeks of contracting
b) Only some strains of the causative agent cause disease in the disease.
humans. d) It is caused by free-living protozoa.
c) Food can taste normal but still cause botulism. e) Distribution of the disease is determined mainly by the
d) Treatment is based on choosing the correct antibiotic. distribution of standing water.
e) Control of the disease depends largely on proper food-canning 10. Which of these statements concerning Creutzfeldt-Jakob disease
techniques. (CJD) and vCJD is true?
5. Which of the following statements about viral meningitis is true? a) CJD occurs in children; vCJD occurs in adults over 45.
a) Vaccines are generally available to protect against the disease. b) CJD and vCJD are sometimes fatal.
b) The main symptom is muscle paralysis. c) CJD is caused by prions; vCJD is a viral infection.
c) Transmission is often by the fecal-oral route. d) Only humans suffer from diseases like CJD and vCJD.
d) The causative agents do not survive well in the e) Both CJD and vCJD produce a spongy appearance in affected
environment. brain tissue.
e) Recovery is rarely complete.
6. Which of these statements concerning arboviral encephalitis is Applications
false? 1. An outbreak of viral meningitis in a small eastern city was linked
a) It is likely to occur in epidemics. epidemiologically to a group who swam a non-chlorinated pool in
b) Mosquitoes can be an important vector. an abandoned quarry outside of town. What might public health
c) Epilepsy, paralysis, and thinking difficulties are among the officials surmise about the probable cause of the outbreak?
possible sequels to the disease. 2. Two microbiologists are writing a textbook, but they cannot agree
d) Use of sentinel chickens helps warn about the disease. where to place the discussion of botulism. One favored the chapter
e) In the United States, the disease is primarily a zoonosis on nervous system infections, whereas the other insisted on the
involving cattle. chapter covering digestive system infections. Where do you think
7. Which of these statements concerning poliomyelitis is false? the discussion should be placed, and why?
a) The sensory nerves are usually involved.
b) It can be caused by any of three specific enteroviruses. Critical Thinking +
c) Only a small fraction of those infected will develop the 1. A pathologist stated that it was much easier to determine the
disease. causative agent of meningitis than of an infection of the skin or
d) The disease is transmitted via the fecal-oral route. intestine. Is her statement valid? Why or why not?
e) A post-polio syndrome can develop years after recovery from 2. Why is it important to learn about rabies when only a few cases
the original illness. occur in the entire United States each year?
27 Blood and Lymphatic Infections
KEY TERMS
Bubo An enlarged, tender lymph
node characteristic of plague and
some sexually transmitted infections.
Disseminated Intravascular
Petechiae Small, purple spots on
the skin and mucous membranes
caused by hemorrhage from small
blood vessels.
Coagulation (DIC) Condition Pneumonic Referring to the lung.
in which clots form in small blood
Sepsis Acute illness caused
vessels throughout the body, causing
by pathogens or their products
organ failure.
circulating in the bloodstream.
Endocarditis Inflammation of the
Septic Shock An array of
heart valves or lining of the heart
effects that results from infection
chambers.
of the bloodstream or circulating
Lymphangitis Inflammation of endotoxin; includes fever, drop in
lymphatic vessels. blood pressure, and disseminated
intravascular coagulation.

victims that he got from British soldiers, whose job it was to bury them.
Koch, p. 548 Kitasato, p. 548
A week after he arrived in Hong Kong, Yersin reported his discov-
SEM of the tip of the mouthparts of a female mosquito. ery of a bacillus that was always present in the swollen lymph nodes of
plague victims. The bacterium could be cultivated, and it caused plague-
like disease when injected into rats. The disease was transmitted from
one rat to another.
A Glimpse of History Kitasato, working with blood from the hearts of plague victims, also
Alexandre Emile John Yersin (1863–1943) is one of the most interesting, announced soon after his arrival in Hong Kong that he had isolated the
though relatively unknown, contributors to the understanding of infectious bacterium that caused plague. It was later proved to be only a laboratory
diseases. He developed an interest in science when, as a young boy, he contaminant, but Kitasato was named co-discoverer of the plague bacillus
found a microscope and dissecting instruments that belonged to his dead because of his great prestige.
father. A local physician befriended Yersin and influenced him to study Yersin’s plague bacillus was named Yersinia pestis. It was used to
medicine. Once he had become a physician, Yersin volunteered at the make a vaccine, and later, antiserum prepared against the organism was
Pasteur Institute in Paris, where he was hired by Emile Roux, a coworker of used to cure a patient with plague—the first successful treatment of a
Louis Pasteur. Yersin became well recognized for his work at the Institute, plague victim. Not long after, another Pasteur Institute scientist proved
but was bored with research, and did not want to practice medicine because that Y. pestis is transmitted by rat fleas.
he felt it was wrong for physicians to make money from other people’s
sickness.

T
Yersin left the Pasteur Institute and was employed as a physician he circulation of blood and lymph fluid supplies nutrients
on a ship sailing from France to Vietnam. He became enchanted with and O2 to cells and carries away their waste products. The
Vietnam and the people who lived there. He made it his home for the rest circulatory system also heats and cools body tissues to
of his life, even establishing a medical school there. maintain an optimum temperature. Infection of the system can
Yersin studied the diseases that affected the Vietnamese people, be serious because infectious agents become systemic, meaning
including plague. The cause and transmission of this serious disease they can be carried to all parts of the body, producing disease in
were completely unknown at the time. It had killed millions in Europe one or more vital organs, or causing the circulatory system itself
in medieval times and had affected France as recently as 1720. In 1894, to stop functioning. When a substance is circulating in the blood-
Yersin went to study an outbreak of plague in Hong Kong. Unfortunately
stream, the condition is given a name that specifies the nature of
for Yersin, Shibasaburo Kitasato, the famous colleague of Robert Koch
(see A Glimpse of History, chapter 23), had arrived 3 days earlier with a
the substance and ends in -emia as in bacteremia, viremia, and
large team of Japanese scientists. British authorities had given Kitasato fungemia. These terms do not imply a disease state—in many
and his colleagues access to patients and laboratory facilities. Yersin cases, the circulating substance does not cause any symptoms. For
did not speak English, so it was difficult for him to communicate his example, a person is often transiently bacteremic after brushing his
requirements to the authorities. He had to set up a laboratory in a bamboo or her teeth, when mouth bacteria enter small abrasions caused by
shack and was forced to do his research on samples from dead plague the brushing. If illness results from a circulating agent or its toxins,

670
 Part IV Infectious Diseases 671

PERSPECTIVE 27.1
Arteriosclerosis: The Infection Hypothesis
Arteriosclerosis, the main cause of heart with it by the time they reach adulthood. What and strokes be tested? Some studies of coro-
attacks and strokes, is characterized by lipid- was interesting about the antibody studies was nary patients treated with antibiotics effective
rich deposits that develop in arteries and can that the higher the titer of antibody, the greater against the bacterium have suggested a benefi-
slow or stop the flow of blood. the risk of coronary disease. Then, in 1996, live cial effect, and others have not. It now appears
In 1988, researchers in Finland reported C. pneumoniae was shown to be present in that many inflammatory conditions, not just
that patients with coronary artery disease— many arteriosclerotic lesions, and other stud- C. pneumoniae infections, increase the risk of
meaning arteriosclerosis of the arteries that ies established that there was an association heart attacks and strokes. The risk correlates
supply the heart muscle—often had antibodies between the bacterium and arteriosclerotic with the level of acute-phase proteins arising
against Chlamydophila pneumoniae (previously lesions of both heart and brain arteries. from the release of pro-inflammatory cyto-
Chlamydia pneumoniae). This organism is a tiny, Does C. pneumoniae cause arterioscle- kines. Undoubtedly there will be more to come
obligate, intracellular bacterium that causes a rosis, does it worsen the effects of arterio- in determining the relationship, if any, between
variety of common respiratory illnesses includ- sclerosis, or does it merely exist harmlessly arteriosclerosis and acute-phase proteins.
ing sinusitis and pneumonia. C. pneumoniae is in the lesions? How can the hypothesis that acute-phase proteins, p. 348
widespread, and most people have been infected C. pneumoniae contributes to heart attacks

the condition is referred to as sepsis, or blood poisoning. Sepsis Blood Vessels

can cause the blood pressure to fall to such low levels that blood
Blood vessels include arteries, veins, and capillaries. Arteries have
flow to vital organs is insufficient to maintain their functioning, a
thick muscular walls to withstand the high pressure of the arter-
condition called septic shock.
ial system. Arterial blood is bright red, the color of oxygenated

27.1 ■ Anatomy, Physiology, Capillaries of head

and upper body
and Ecology
Learning Outcome
1. Describe the characteristics and functions of the heart, blood Right lung Left lung
vessels, lymphatics, and spleen. capillaries capillaries

The cardiovascular system is composed of the heart, blood ves-

sels, and blood. The lymphatic system consists of lymph, lymph
vessels, lymph nodes, and lymphoid organs, including the tonsils,
appendix, and spleen (figure 27.1). Both systems are normally
sterile. As blood flows around the body, it passes alternately
through the lungs and tissue capillaries. During each circuit, some
of the blood passes through the lymphoid organs, which contain
phagocytic cells that remove infectious agents and other foreign
material. lymphatic system, p. 357
Pericardium
Pericarditis Left atrium
Left ventricle
The Heart Heart muscle
Myocarditis
The heart—a muscular pump enclosed in a fibrous sac called the
pericardium—moves the blood around the body. It is divided into Lymphatic Lymph Valves
system node Endocarditis
right and left sides by a septum, a wall of tissue through which Lymphangitis
blood normally does not pass after birth (figure 27.1). The right
and left sides of the heart are each divided into two chambers—the Venous Arterial
system system
atria, which receive blood, and the ventricles, which discharge it.
Blood from the right ventricle flows through the lungs and into
the left atrium. From there, the blood passes into the left ventricle
and is then pumped through the aorta to the arteries and capillar- Capillaries of body
ies that supply the tissues of the body. The blood is then returned organs and other tissues
to the right atrium by the veins. Valves at the entrance and exit of FIGURE 27.1 The Blood and Lymphatic Systems Disease
each ventricle prevent the blood flow from reversing. Although conditions are indicated in red type. For simplicity, the spleen is not
not common, infections of the heart valves, heart muscle, and shown.
pericardium can be serious because they affect blood circulation. ? Which chambers of the heart receive blood, and which discharge it?
672 Chapter 27 Blood and Lymphatic Infections

hemoglobin. Infection of the arteries is unusual, although the aorta lysozyme, and interferon. An inflammatory response may cause
can be dangerously weakened in people with syphilis. Arteries may lymph and blood to clot in vessels close to areas of infection, pre-
become affected by arteriosclerosis (lipid-rich deposits that block venting microbes from spreading. leukocytes, p. 339 antimicrobial
the vessels), putting people at risk for heart attacks and strokes. substances, p. 337 inflammatory response, p. 348
Some studies suggest that bacteria and viruses play a role in arterio-
sclerosis (see Perspective 27.1). syphilis, p. 626
Venous blood is dark red, because it becomes depleted of Spleen
O2 in the capillaries. Valves in veins play an important role in The spleen is a fist-sized organ found on the left side of the
keeping the blood flowing in one direction, because the venous abdominal cavity, behind the stomach. It contains two kinds of
pressure is too low to do this. Veins are easily compressed, so the tissue—red pulp (multiple blood-filled passageways) and white
action of muscles also aids the flow of venous blood. pulp (lymphoid tissue). The spleen has several functions. The red
pulp cleans the blood by filtration, in the same way that the lymph
nodes clean the lymph. Large numbers of phagocytes in red pulp
Lymphatics (Lymphatic Vessels)
remove aging or damaged red blood cells (RBCs), bacteria, and
The lymphatic system consists of lymphatic vessels that resemble other foreign materials from the blood. It also produces new blood
blood capillaries but are larger (see figure 15.3). The vessels carry cells in rare situations where the bone marrow is unable to make
an almost colorless fluid called lymph, which comes from plasma, enough, and has a reserve of monocytes. The white pulp, which
the non-cellular portion of the blood. Lymph fluid seeps through contains both B and T lymphocytes, provides an active immune
the walls of the blood capillaries to become the interstitial fluid response to microbial invaders.
that surrounds tissue cells. This fluid bathes and feeds the tissue
cells and then enters the lymphatics (see figure 15.4). Unlike the MicroAssessment 27.1
blood capillaries, lymphatic vessels are easily permeable and take
up foreign material such as invading microbes and their products, The heart has four chambers that work together to move the blood
around the body through arteries and veins. The lymphatic system
including toxins and other antigens. Many one-way valves in the
cleans interstitial fluid and returns it to the bloodstream. Systemic
lymphatic vessels keep the flow of lymph moving away from the infections may disrupt the transport of O2 and nutrients to body
lymphatic capillaries. Lymph fluid is moved by both contraction tissues and removal of waste products. Infectious agents and their
of the vessel walls and compression by the body’s muscles. toxins can be spread throughout the body by the circulatory system.
Lymphatic vessels drain into multiple lymph nodes. These 1. What are the functions of the spleen?
nodes, which contain phagocytic cells and lymphocytes, are 2. What is the function of valves in the lymphatic and circulatory
where foreign materials such as microbial cells are trapped and systems?
destroyed. Lymph flows out of the nodes through vessels that 3. Which side of the heart—right or left—do bacteria in infected
eventually combine into one large tube—the thoracic duct. This lymph reach first? Where do they go from there? +
duct then empties into a large vein (the subclavian vein) behind the
left collarbone, and back into the main blood circulation. When a
hand or a foot is infected, a visible red streak may spread up the
limb from the infection site toward the nearest lymph node, a con- 27.2 ■ Bacterial Diseases of the
dition called lymphangitis (figure 27.2). phagocytic cells, p. 346 Blood Vascular System
Blood and lymph both carry infection-fighting leukocytes
and antimicrobial proteins including antibodies, complement, Learning Outcomes
2. Outline the events that lead to subacute bacterial endocarditis.
3. Explain the role of the inflammatory response in sepsis and
septic shock.

Bacterial infections of the vascular system can kill a person quickly,

or they can progress slowly for months, causing a gradual decline in
health. They are not common, but they are always dangerous. Usually
the bacteria access the bloodstream as they are carried by lymph flow-
ing from an infected area in the tissues. Some pathogens multiply in the
bloodstream, and they may colonize and form biofilms on structures
such as the heart valves. biofilm, p. 84
Endocarditis is the term used for infections of the heart valves
or the inner surfaces of the heart. Acute bacterial endocarditis
starts suddenly with fever and is usually caused by virulent
species such as Staphylococcus aureus and Streptococcus
pneumoniae, which can infect both normal and abnormal heart
FIGURE 27.2 Lymphangitis Notice the red streak extending up valves. These bacteria quickly destroy the valves and form abscesses
the arm. in the heart muscle, causing heart failure. By contrast, subacute
? What causes lymphangitis? bacterial endocarditis is usually caused by organisms with little
 Part IV Infectious Diseases 673

virulence, progresses much slower, and is less likely to be fatal. prophylactic antibiotic treatment before dental procedures to patients
Staphylococcus aureus, p. 524 Streptococcus pneumoniae, p. 497 with serious heart murmurs. A heart murmur is an abnormal sound
Sepsis is caused by both Gram-negative and Gram-positive the physician hears when listening to the heart, often indicative of a
bacteria, as well as other infectious agents. deformed valve or other structural abnormality.
SBE also occurs in injected-drug abusers, hospitalized patients
Subacute Bacterial Endocarditis (SBE) who have plastic intravenous catheters for long periods, and those
Subacute bacterial endocarditis (SBE) is usually localized to one with artificial heart valves. These people are usually infected with
of the valves on the left side of the heart. It commonly occurs on S. epidermidis, S. aureus, enterococci, or a wide variety of species
valves that are deformed because of a birth defect or a disease other than viridans streptococci.
such as rheumatic fever. rheumatic fever, p. 489
Treatment and Prevention
Signs and Symptoms SBE is treated with antibacterial medications chosen according
People with SBE usually have noticeable fatigue and slight fever. to the susceptibility of the causative organism. Only bactericidal
They typically become ill gradually and slowly lose energy over a medications such as penicillin and gentamicin are effective, and
period of weeks or months. They may suddenly develop a stroke. usually two or more antimicrobials are used together. Prolonged
treatment over one or more months is usually required. Infection
Causative Agent of an artificial heart valve or other foreign material is almost
SBE is usually caused by normal bacterial microbiota of the always associated with biofilm formation, and the object must
mouth or skin, such as viridans streptococci and Staphylococcus often be replaced to cure the person. Sometimes, surgery is needed
epidermidis. The infecting organisms are usually shed from to remove an infected clot or to drain abscesses.
the infected heart valve into the blood and can be identified by There are no proven methods for preventing SBE. However,
cultivating samples taken from an arm vein. However, it is not people with known or suspected heart valve abnormalities are
always possible to culture the causative agent. For example, blood commonly given an antibacterial medication shortly before den-
that has passed through the lung before reaching the arm vein is tal or other bacteremia-causing procedures. The medication is
cleared of microorganisms by lung phagocytes. Also, some patho- chosen according to the expected bacterial species and its likely
genic bacteria that cause endocarditis, such as Coxiella burnetii, susceptibility to antimicrobials. Healthcare-associated SBE can
cannot be cultured in cell-free media. In these cases, the causative be prevented by strict use of sterile technique when inserting
agent is identified by PCR and nucleic acid probes. Coxiella plastic intravenous catheters, moving the catheters to a new site
burnetii, p. 277 PCR, p. 227 nucleic acid probes, p. 232 every few days, and removing them as soon as possible. Such
precautions help prevent bacterial colonization of the catheters
Pathogenesis and consequent bacteremia that could lead to heart valve infec-
The bacteria that cause SBE enter the blood during dental proce- tion. The main characteristics of SBE are presented in table 27.1.
dures, tooth brushing, or trauma. These microbes can get trapped in healthcare-associated infections, p. 449
the thin blood clots that often form around deformed heart valves or
other areas with disturbed blood flow. They multiply there, creating
a biofilm that protects them from phagocytosis and antimicrobial TABLE 27.1 Subacute Bacterial Endocarditis
medications. The clot grows larger around the multiplying organ- Signs and Fever, loss of energy over a period of weeks
isms, gradually building up a fragile mass. Bacteria continually symptoms or months; sometimes, a stroke
wash off the mass into the circulation, and pieces of infected clot Incubation period Poorly defined, usually weeks
(septic emboli) can break off. These can block important blood ves-
Causative agents Usually oral α-hemolytic viridans streptococci
sels, leading to death of the tissue supplied by the vessel. They can
or Staphylococcus epidermidis
also cause a vessel to weaken and balloon out, forming an aneurysm.
Pathogenesis Normal microbiota enter bloodstream through
People with SBE often have high levels of antibodies against
dental procedures, other trauma; in an
the bacteria. These may be harmful because they lead to the for- abnormal heart, turbulent blood flow causes
mation of immune complexes, which may lodge in the skin, eyes, formation of a thin clot that traps circulating
and other body structures, triggering an inflammatory response. organisms; a biofilm forms, protecting the
In the kidney, the complexes cause glomerulonephritis. immune organisms from phagocytic killing; pieces of
clot break off, block important blood vessels,
complexes, p. 395 glomerulonephritis, p. 490
leading to tissue death.
Even though the bacteria that cause SBE normally have little
Epidemiology People at risk are mainly those with
invasive ability, large numbers of them growing in the heart are
congenital heart defects or with hearts
sometimes able to penetrate into heart tissue, producing abscesses damaged by disease such as rheumatic fever;
or damaging valve tissue, and resulting in a leaky valve. may develop after dental procedures or
other situations that cause bacteremia.
Epidemiology Treatment Treatment: bactericidal antibiotics given
In recent years, viridans streptococci have caused fewer cases of and prevention together, such as penicillin and gentamicin.
SBE cases than previously. Cases that do occur, however, are more Prevention: giving an antibiotic immediately
serious because the causative organisms are becoming increasingly before anticipated bacteremia, such as before
dental work.
antibiotic-resistant. This may be an unintended result of giving
674 Chapter 27 Blood and Lymphatic Infections

Sepsis and Septic Shock compounds recruit additional phagocytes from the bone mar-
row and cause the phagocytic cells to produce more TLRs. This
Sepsis, an infection-induced systemic inflammatory response, is
increases the system’s sensitivity to PAMPs, leading to an even
a common healthcare-associated illness. The disease is caused by
greater release of pro-inflammatory mediators. The phagocytic
the release of bacterial products—particularly endotoxins from
cells and dying host cells then release compounds that function
Gram-negative bacteria—that change the normally beneficial
as damage-associated molecular patterns (DAMPs). The host
inflammatory response into an excessive damaging response that
responses to PAMPs and DAMPs leads to multiple devastating
can lead to life-threatening illness. If uncontrolled, sepsis can
outcomes, including the inhibition of systems that normally control
progress to septic shock, a dramatic drop in blood pressure. The
inflammation, suppression of the adaptive immune response, and
discovery that sepsis involves pro-inflammatory cytokines and
activation of the coagulation cascade (figure 27.3). DAMPs, p. 342
not only invading microorganisms has led to advances in under-
The net result of this dysregulated inflammatory response is
standing the pathogenesis of this disease. endotoxic shock, p. 394
complex and varied. Activation of the coagulation cascade causes
pro-inflammatory cytokines, p. 341
small clots to form in the capillaries. This blocks them, cutting
MicroByte off the blood supply to tissues, which leads to tissue hypoxia
An estimated 400,000 cases of Gram-negative sepsis occur in the (low levels of O2) and subsequent tissue necrosis (death). The
United States each year. widespread clotting—disseminated intravascular coagulation
(DIC)—is often accompanied by hemorrhage, because the endo-
thelium is damaged. DIC may lead to multiorgan failure due to
Signs and Symptoms circulation disruptions. At the same time, phagocytes release
The signs and symptoms of severe sepsis include violent shak- tissue-damaging lysosomal enzymes. This is particularly impor-
ing, chills, and fever, often with rapid breathing and feelings of tant in the lungs, which are seriously and irreversibly damaged
anxiety. If septic shock develops, urine output drops, respiration by these enzymes. Lung damage often results in death even if
and pulse speed up, and the arms and legs become cool and dusky- the individual’s infection has been cured. The harmful effects of
colored. Septic patients are often referred to as “looking ill.” the inflammatory response are made worse by the hypotension
(low blood pressure) usually present. The hypotension is caused
Causative Agents by decreased muscular tone of the heart and blood vessel walls
Systemic infection by any microorganism can cause sepsis, but most
fatal cases involve Gram-negative bacteria. Recall that the outer
membrane of Gram-negative bacteria contains lipopolysaccharide
(LPS), also referred to as endotoxin (see figure 3.33). Examples
Shock
of common causes of fatal sepsis include members of the normal Impaired O2 exchange
microbiota of the large intestine—particularly facultative anaer-
obes such as Escherichia coli and other Enterobacteriaceae mem- Decreased Increased Increased leakage
muscle tone of adhesiveness of plasma from
bers—and anaerobes such as Bacteroides species. Environmental heart and arteries Fever of neutrophils blood vessels
bacteria such as Pseudomonas aeruginosa are also common causes.
lipopolysaccharide, p. 60 Enterobacteriaceae, p. 265 Pseudomonas, p. 265
Cytokines released
Pathogenesis
Macrophages
Sepsis almost always starts from an infection somewhere in the activated
body other than the bloodstream—a kidney infection, for example.
When normal body defenses are compromised by medical treat-
Endotoxin from
ments such as surgery, catheters, and some medications, microor- Gram-negative
Complement Clotting
ganisms that normally have little invasive ability are able to infect activated bacteria released into activated
the blood. bloodstream
Sepsis progresses in stages and is initially due to an overstimu-
Complement Disseminated
lation of the inflammatory response. When the toll-like receptors components intravascular
(TLRs) on macrophages and neutrophils detect endotoxin or other coagulation
pathogen-associated molecular patterns (PAMPs), the phagocytes
respond by releasing pro-inflammatory cytokines. When this occurs
systemically, an uncontrolled release of cytokines—a cytokine Leukocytes Increased Lysosomal Depletion Tissue
attracted to capillary enzymes of clotting damage
storm—results. The complement pathway is also activated, further lung tissue leakage released from proteins from clots
amplifying the inflammatory response. toll-like receptors, p.  342 of plasma leukocytes in capillaries
PAMPs, p. 342 cytokines, p. 341 complement system, p. 344
Lung tissue damage Hemorrhage
The systemic release of pro-inflammatory cytokines,
along with activation of complement, results in a widespread, FIGURE 27.3 Events in Gram-Negative Sepsis
self-stimulating inflammatory response. Various pro-inflammatory ? What causes organ failure in sepsis?
 Part IV Infectious Diseases 675

and the low blood volume that results from fluid leakage out MicroAssessment 27.2
of the blood vessels. Current evidence indicates that the pro-
inflammatory cytokines lead to septic cardiomyopathy (disorder Bacteria of low virulence can cause serious, even fatal, infections
in individuals with a structural abnormality of the heart. A systemic
of the heart muscles) and heart failure. Shock results when the infection represents failure of the body’s mechanisms for keeping
blood pressure falls so low that vital organs are no longer supplied infections localized to one area. The inflammatory response, although
with adequate amounts of blood to maintain their function. vitally important in localizing infections, can be life-threatening if
Even if the patient survives the initial stages of sepsis, the generalized. People die of Gram-negative sepsis despite antibacterial
later effects can be fatal. Neutrophils stop working properly, so therapy because of the endotoxin-mediated release of cytokines in the
bacteria are not cleared from the blood effectively. In addition, lung and bloodstream.
lymphocytes—particularly helper T cells—and dendritic cells 4. What is a systemic infection?
undergo apoptosis, affecting the adaptive immune response and 5. What is the difference between bacteremia and sepsis?
leading to immunosuppression that results in increased suscepti- 6. Why might clots on the heart valves make microorganisms there
bility to secondary infection. inaccessible to phagocytic killing? +

Epidemiology
Sepsis is mainly a healthcare-associated disease, reflecting the 27.3 ■ Bacterial Diseases of the
high incidence of bloodstream infections in hospitalized patients
with impaired host defenses. Rates of sepsis have been increasing,
Lymph Nodes and Spleen
likely due to longer life spans, antibiotic suppression of normal
Learning Outcome
microbiota, and immunosuppressive medications. Use of medical
4. Compare and contrast tularemia, brucellosis, and plague.
equipment where biofilms readily develop—such as respirators,
and catheters placed in blood vessels and the urinary system—also
Enlargement of the lymph nodes and spleen is a prominent feature of
increases the risk of sepsis.
diseases that involve the mononuclear phagocyte system. The exam-
Treatment and Prevention ples discussed in this section are now uncommon human diseases in
the United States, but they represent a constant threat because they
Treatment of sepsis is difficult, because it is a complex syn-
are widespread in animals. mononuclear phagocyte system, p. 340
drome involving several host responses. It is currently treated
with antimicrobial medications effective against the causative
organism. Bactericidal antibiotics often lyse bacterial cells, how- Tularemia (“Rabbit Fever”
ever, increasing the release of endotoxin from Gram-negative or “Deer Fly Fever”)
organisms, worsening the effects of the sepsis. The person is also Tularemia is widespread in the United States and is found in wild
treated for shock and tissue hypoxia. In addition, fluid replace- animals such as rabbits, muskrats, and bobcats. The disease was
ment and support for organ dysfunction are given. identified in Tulare County, California, following an outbreak
New treatments for sepsis are being evaluated. Monoclonal among ground squirrels in the area in 1911. Although the disease
antibodies against endotoxin or pro-inflammatory cytokines affects humans only occasionally in the United States today, it is
such as tumor necrosis factor (TNF) can have some benefit when potentially a bioterrorism threat.
given before shock develops. Another medication being evalu-
ated is drotrecogin alfa (activated human protein C produced by Signs and Symptoms
recombinant technology), which opposes the effects of certain Signs and symptoms of tularemia depend on how the causative
pro-inflammatory cytokines. Evaluation of new treatments organism enters the body—through mucous membranes or minor
is difficult because (1) many of the patients have underlying cuts and scratches in the skin, or by inhalation. Typically, signs
life-threatening illnesses in addition to their sepsis, and (2) ani- and symptoms appear 2 to 5 days after a person is bitten by an
mal models for sepsis do not accurately mimic the disease in infected tick or insect or handles an infected wild animal. The
humans. Other possible targets for new therapies include TLR4 characteristic sign is an ulcer that develops at the site where the
(the TLR that recognizes LPS), complement component C5a, organism enters the body (figure  27.4). Regional lymph nodes
and the autonomic nervous system, which plays a regulatory enlarge, and fever, chills, and achiness occur. Signs and symptoms
role in inflammation by controlling release of pro-inflammatory usually clear in 1 to 4 weeks, but sometimes they last for months.
cytokines. Between 30% to 50% of people with sepsis die even If the organism is inhaled or infects the lung from the blood-
with treatment, partly because of their other underlying illnesses. stream, pneumonia may occur. Signs and symptoms include a
monoclonal antibodies, p. 428  dry cough and pain beneath the sternum (breastbone), a result of
Sepsis can be prevented by quick identification and effec- enlarged lymph nodes. Tularemic pneumonia, although rare, is
tive treatment of localized infections, particularly in people with more serious and has a mortality rate as high as 30% if untreated.
impaired immune defenses. Also, predisposing conditions such
as bedsores and pyelonephritis—which commonly lead to sepsis Causative Agent
in patients with cancer and diabetes—can usually be prevented. Tularemia is caused by Francisella tularensis, a non-motile,
pyelonephritis, p. 613 aerobic, Gram-negative rod. It was named for Edward Francis, an
676 Chapter 27 Blood and Lymphatic Infections

Treatment and Prevention

Most cases of tularemia are effectively treated with streptomycin.
Ciprofloxacin or gentamicin can also be used.
Tularemia can be prevented by using rubber gloves and
goggles or face shields when skinning or handling wild animals.
Insect repellants and protective clothing help protect against insect
and tick vectors. The disease can also be avoided by removing
ticks found after outdoor activity. Meat from wild animals should
be cooked thoroughly. The main features of this disease are sum-
marized in table 27.2.

Brucellosis (“Undulant Fever”

or “Bang’s Disease”)
Brucellosis is often called “undulant fever,” or “Bang’s disease,”
after Frederik Bang (1848–1932), a Danish veterinary professor
FIGURE 27.4 Tularemic Ulcer of the Thumb This kind of lesion who discovered the cause of cattle brucellosis. Only about 150
is generally acquired through skinning rabbits or other wild animals.
cases of human brucellosis are reported each year in the United
? In what other ways can tularemia be transmitted? States. Many more cases, however, go unreported annually.

Signs and Symptoms

Brucellosis usually starts gradually, and the signs and symptoms
American physician who studied tularemia in the early 1900s,
are vague. Typically, patients complain of mild fever, sweating,
and Tulare County, where it was first identified. The organism is
weakness, aches and pains, enlarged lymph nodes, depression and
unusual because it requires a special medium enriched with the
weight loss. The recurrence of fevers over weeks or months in
amino acid cysteine in order to grow.
some cases gave rise to the alternative name, “undulant fever.”
Most people recover within 2 months even without treatment, and
Pathogenesis
only 15% are symptomatic for more than 3 months.
Francisella tularensis enters through breaks in the skin or mucous
membranes and is carried to the regional lymph nodes, which Causative Agent
become large and tender. The nodes may become filled with pus Brucella sp. are small, aerobic, nonmotile, Gram-negative rods with
and drain spontaneously. The organism spreads to other parts of complex nutritional requirements. The causative agent was discov-
the body via the lymphatics and blood vessels. F. tularensis is ered by Dr. David Bruce, after whom it was named. Four varieties of
ingested by phagocytic cells and grows within them. This may
explain why the disease persists in some people despite the high
antibody titers in their blood. Cell-mediated immunity effectively Tularemia (“Rabbit Fever”
rids the host of this infection. Even without treatment, over 90% TABLE 27.2
or “Deer Fly Fever”)
of infected people survive. cell-mediated immunity, p. 356
Signs and Ulcer at site of entry, enlarged lymph nodes
symptoms in area, fever, chills, achiness
Epidemiology
Incubation period 1 to 10 days; usually 2 to 5 days
Tularemia occurs among wild animals in many areas of the
Northern Hemisphere, including all the states of the United States Causative agent Francisella tularensis, an aerobic,
except Hawaii. In the eastern United States, summertime human Gram-negative rod
infections commonly result from tick bites. However, cases often Pathogenesis Organisms are ingested by phagocytic cells,
occur in the winter months as well, in people skinning rabbits grow within these cells, and then spread
throughout body.
(explaining the common name, “rabbit fever”). People also con-
tract the disease from muskrats, beavers, squirrels, deer, and other Epidemiology Present among wildlife in most states of the
United States. Risk mainly to hunters, game
wild animals. The animals are generally free of illness but carry the
wardens, and others who handle wildlife.
causative organism. In the western United States, infections mostly Acquired when the organism penetrates
result from the bites of infected ticks and deer flies (explaining the a mucous membrane or enters broken skin,
other common name, “deer fly fever”) and usually occur during as may occur when skinning rabbits,
summer. Generally, 150 to 250 cases of tularemia are reported for example; bite of infected insect
or tick.
each year from counties across the United States. Epidemics of
inhalation tularemia have occurred from dust arising from mow- Treatment Treated with gentamicin or ciprofloxacin.
and prevention Prevention: avoiding bites of insects and
ing lawns or from rodent-infested buildings. Tularemia can also
ticks; wearing rubber gloves, goggles, when
be contracted by ingesting contaminated material. In the United skinning rabbits; taking safety precautions
States, F. tularensis is listed among the category A (highest-risk) when working with organisms in laboratory.
agents of biological terrorism (see Future Challenges 19.1).
 Part IV Infectious Diseases 677

the genus Brucella cause brucellosis in humans. DNA studies show Most brucellosis in humans is associated with occupational
that all members of the genus fall into a single species, Brucella exposure—workers in the meat packing industry, veterinarians,
melitensis, but traditionally, the different varieties were assigned and slaughterhouse workers can be exposed to the causative
species names depending largely on their preferred host: B. abortus agents. People drinking unpasteurized milk or eating soft cheeses
invades cattle; B. canis, dogs; B. melitensis, goats; and B. suis, pigs. made from infected milk can also contract the disease. In the
The distinctions between the various strains are mainly useful epide- United States, hunters have acquired infections from elk, moose,
miologically and generally have little pathogenic significance. bison, caribou, and reindeer. B. melitensis is listed as a category B
(intermediate-risk) bioterrorism agent.
Pathogenesis
Brucella organisms penetrate mucous membranes or breaks in MicroByte
the skin and are disseminated via the lymphatic and blood vessels About 20% of the Yellowstone Park bison carry brucellosis, and
to the heart, kidneys, and other parts of the body. The infection over 1,000 have been killed when they wandered from the park, in
causes spleen enlargement. Like Francisella tularensis, Brucella order to prevent transmission of the disease to cattle.
sp. are not only resistant to phagocytic killing but also can grow
intracellularly in phagocytes, where they are inaccessible to anti- Treatment and Prevention
bodies and some antibiotics. The mortality rate is low and death is Brucellosis can usually be treated using doxycycline with rifampin
generally due to endocarditis. A serious but rare complication is or streptomycin for 6 weeks. In some chronic cases, extended
bone infection (osteomyelitis). treatment (sometimes up to 6 or more months) may be needed.
The most important control measures for brucellosis are pas-
Epidemiology teurization of dairy products and inspection of domestic animals
Brucellosis is typically a chronic infection of domestic animals. It for evidence of the disease. Veterinarians, butchers, and slaugh-
affects the mammary glands and the uterus, contaminating milk and terhouse workers should use protective gear such as goggles or
causing abortions in the affected animals (abortion is not a feature of face shield and rubber gloves. An attenuated vaccine effectively
human disease). Worldwide, brucellosis is a major problem in ani- controls the disease in domestic animals. Table  27.3 gives the
mals used for food, causing yearly losses of many millions of dollars. main features of brucellosis.

TABLE 27.3 Brucellosis (“Undulant Fever” or “Bang’s Disease”)

1 Brucella melitensis enters Signs and Fever, body aches, weight loss, enlargement
the body through mucous symptoms of lymph nodes; symptoms may subside
membranes, skin abrasions, 1 without treatment but then recur
or ingestion of unpasteurized 1
Incubation period Usually 5 to 21 days
milk.
Causative agent Brucella melitensis, a small, aerobic,
2 The bacteria are taken up by
Gram-negative rod
phagocytes but resist digestion
and grow within cells. Pathogenesis Organisms penetrate mucous membranes
3
4 and are carried to heart, kidneys, and
3 The bacteria enter the 5 other parts of the body via the blood and
lymphatics and bloodstream
lymphatic system; they are resistant to
and are carried throughout the
phagocytic killing and grow within these
body.
4 cells.
4 Infection is established in other
Epidemiology Main sources of human infections: domestic
body tissues, such as the heart
animals. Disease also occurs in wild animals
valves, kidneys, and bones.
such as moose, caribou, bison; butchers,
5 Osteomyelitis is the most farmers, veterinarians, those who drink
common serious complication. 1 unpasteurized milk are at risk.
Most deaths are due to 4
Treatment Treatment: doxycycline with rifampin or
endocarditis.
and prevention streptomycin. Prevention: vaccination
5 of domestic animals; pasteurization of
milk and milk products; gloves and face
protection.
678 Chapter 27 Blood and Lymphatic Infections

Plague (“Black Death”) FIGURE 27.6 Fleas

Following a Blood Meal
Plague, once known as the “black death,” was responsible for (a) Healthy flea with fresh
the death of approximately a quarter of the population of Europe blood in gut. (b) Flea with
between 1346 and 1350. Crowded conditions in the cities and a obstruction due to Yersinia
pestis infection.
large rat population played major roles in the spread of the disease.
Plague is a potential bioterrorism disease, listed as category A. ? What is the risk when (a)
a flea has a digestive
Signs and Symptoms system obstruction?

The signs and symptoms of plague depend on how the disease was
acquired. Most commonly, a person is bitten by an infected flea,
and symptoms develop 2 to 6 days later. The person characteristi-
cally develops markedly enlarged and tender lymph nodes called
buboes—hence the name bubonic plague—in the region that
receives lymph drainage from the area of the flea bite. High fever,
shock, delirium, and patchy bleeding under the skin quickly develop.
If a person inhales respiratory droplets from an infected patient (b)
or animal, they may develop pneumonic plague with signs and discharge bacterial cells in their saliva, and these can then be
symptoms of cough and bloody sputum. Pneumonic symptoms introduced into human tissue when a person scratches the flea bite.
arise more quickly than bubonic symptoms, typically in 1 to 3 days. Y. pestis has multiple virulence factors, making it extremely
If the causative organisms spread via the bloodstream, the well equipped for avoiding mammalian host defenses. Some
person may develop septicemic plague. Endotoxin released by virulence factors are encoded on the bacterial chromosome and
the causative agent results in shock and disseminated intravascular others are encoded by various plasmids. The characteristics of the
coagulation (DIC), which causes bleeding into the skin and organs, most important virulence factors are summarized in table 27.4.
leading to a red or black patchy rash. DIC, p. 674 Once Y. pestis enters via a flea bite, its protease (Pla) clears the
lymphatics and capillaries of clots and inactivates certain complement
Causative Agent system components, allowing the organisms to spread. The lymphatic
Plague is caused by Yersinia pestis, which, like other members of vessels carry the bacterial cells to the regional lymph nodes, where
the Enterobacteriaceae, is a facultatively anaerobic, Gram-negative they are taken up by macrophages. The bacteria sense the intracellular
rod. It is non-motile and grows best at 28°C. The organism resem- conditions of the macrophages and respond by turning on various
bles a safety pin when stained with certain dyes because the ends genes required for surviving in the mammalian host. Some of these
of the bacterium stain more intensely than the middle (figure 27.5). allow the organisms to resist the killing effects of the macrophages
and to multiply within them. After several days, an acute inflamma-
Pathogenesis tory reaction develops in the lymph nodes, producing the enlargement
Yersinia pestis forms biofilms in the digestive tract of infected and marked tenderness that characterizes bubonic plague.
fleas, often blocking the tract (figure  27.6). This both starves The infected macrophages eventually die and release the
the flea—thereby increasing the likelihood that it will try to bacteria, which are now “armed” to withstand host defenses. For
feed again—and causes bacteria to be regurgitated into the bite example, the Yersinia outer membrane proteins (YOPs) and capsule
wound as the flea attempts to feed. Even fleas that do not have allow Y. pestis to avoid phagocytosis, and iron acquisition proteins
an obstructed digestive tract can transmit Y. pestis because they allow it to trap hemin. The lymph nodes may become necrotic,
allowing large numbers of the bacterial cells to spread into the
bloodstream, causing septicemic plague. Endotoxin released by the
organisms causes the signs and symptoms of septicemic plague,
including shock and DIC. The dark hemorrhages and dusky color
of skin and mucous membranes from DIC probably inspired the
common name “black death.”
In 10% to 20% of the cases, the infection spreads to the lungs,
resulting in pneumonic plague. A person with pneumonic plague
can transmit the disease to others in respiratory droplets. Organisms
acquired this way are already fully virulent and, therefore, are espe-
cially dangerous.

Epidemiology
Plague is endemic in rodent populations worldwide except Australia.
In the United States, the disease mostly occurs in about 15 states
5 μm in the western half of the country. The main reservoirs are prai-
FIGURE 27.5 Yersinia pestis rie dogs and rock squirrels, as well as their fleas. However, rats,
? What is the function of the Yops proteins made by this organism? rabbits, dogs, and cats can also be hosts. Hundreds of species of
 Part IV Infectious Diseases 679

plague patients. The number of reported cases of plague is gradu-

Virulence Factors
TABLE 27.4 ally increasing as cities spread into surrounding areas where the
of Yersinia pestis
main reservoirs live. African countries have the highest number of
Factor Coded by Action cases worldwide, but the disease is endemic in 10 Chinese prov-
Pla pPCP1 Activates plasminogen; destroys
inces, where earthquakes create conditions for pneumonic plague.
(protease) plasmid C3b, C5a and clots endemic disease, p. 438 reservoir, p. 438

YOPs pCD1 Interfere with phagocytosis and

(proteins) plasmid the immune response Treatment and Prevention
V antigen pCD1 Controls type III secretion system Plague can be effectively treated with antimicrobial medications
plasmid that delivers YOPs proteins including gentamicin, ciprofloxacin, or doxycycline, especially if
given within 24 hours of the onset of signs and symptoms. Between
F1 pMT1 Forms antiphagocytic capsule at
plasmid 37°C
50% and 80% of people with bubonic plague die if not treated.
Mortality rate for untreated pneumonic plague is almost 100%.
PsaA Chromosome Role in attachment to host cells
(adhesin)
Plague epidemics can be prevented by rat control measures
such as proper garbage disposal, constructing rat-proof buildings,
Complement Chromosome Protects against lysis by activated
resistance complement
and rat extermination programs. These programs must be com-
bined with the use of insecticides to prevent the escape of infected
Iron Chromosome Traps hemin and other iron-
acquisition containing substances; stores
fleas from dead rats. There is no current vaccine for plague, but
iron compounds intracellularly new genetically engineered vaccines are being evaluated under a
joint agreement among the United States, Canada, and the United
Kingdom. The antibiotic tetracycline can be given as a preven-
fleas are able to transmit plague, and the fleas can remain infectious tive for someone exposed to plague and is useful in controlling
for a year or more. Epidemics of pneumonic plague in humans are epidemics because of its immediate effect. The main features of
caused by respiratory droplets produced by coughing pneumonic plague are presented in table 27.5.

TABLE 27.5 Plague (“Black Death”)

1 Yersinia pestis is acquired from Signs and Sudden onset of high fever, large lymph
the bite of an infected flea or symptoms nodes called buboes, skin hemorrhages;
scratching skin contaminated sometimes bloody sputum
by the flea’s feces. Incubation period Usually 1 to 6 days
2 The bacteria are carried to 7 Causative agent Yersinia pestis, a Gram-negative rod; a
regional lymph nodes. member of the Enterobacteriaceae
3 Phagocytes ingest the bacteria with multiple chromosome- and
but the intracellular conditions plasmid-coded virulence factors
activate capsule production Pathogenesis Enters the body with bite of infected
and other genes responsible 5 flea. Bacteria taken up by macrophages.
6
for virulence. Intracellular environment causes
4 Fully virulent bacteria break them to transform into encapsulated
out of the phagocytes, infect organisms capable of producing
the nodes, producing the multiple virulence factors that allow
buboes that characterize attachment to host cells, and provide
bubonic plague. 2 defense against the immune system.
5 The bacteria may be carried Epidemiology Endemic in rodents and other wild
into the bloodstream, 4 3 animals, and their fleas, particularly in
causing septicemic plague. the western states of the United States.
Bubonic plague is transmitted by fleas;
6 The lungs can become
pneumonic plague can be transmitted
infected, producing the
person to person in respiratory
highly contagious and lethal
droplets. Pneumonic plague is the most
pneumonic plague.
dangerous because Y. pestis is fully
7 Bacteria exit with coughing. virulent at the time of transmission.
Treatment Treatment: prompt diagnosis and
and prevention antibacterial treatment necessary to
1 prevent high mortality. Prevention:
currently no vaccine available; new
vaccines are under evaluation. Avoid
contact with wild rodents and their
burrows. Insecticides and rat control.
680 Chapter 27 Blood and Lymphatic Infections

MicroAssessment 27.3 It is a double-stranded DNA virus of the herpesvirus family, and

although identical in appearance to the other known herpesviruses
Generally, tularemia is a bacteremic disease of wild animals transmitted that cause human disease, it is not closely related to any of them.
to humans by exposure to their blood or tissues, or by biting ticks and
insects. Brucellosis is most commonly a disease of domestic animals MicroByte
transmitted to humans when individuals handle the animals’ flesh or EBV, the cause of infectious mononucleosis, was discovered in the
drink unpasteurized milk. Plague is endemic in rodents in the western 1960s when it was isolated from Burkitt’s lymphoma, a malignant
United States and is transmitted to humans by flea bites. Unlike tumor derived from B lymphocytes.
tularemia and brucellosis, plague has the potential to spread rapidly
from human to human by coughing, with a high fatality rate.
7. Workers in what industry are especially likely to contract
Pathogenesis
brucellosis? EBV initially infects the mouth and throat and then becomes
8. How would growth within phagocytes protect Francisella latent in another cell type (figure 27.7). After replicating in the
tularensis from destruction by cell-mediated immunity? mouth epithelium, salivary glands, and throat, the virus is carried
9. How would crowded conditions in cities favor spread to the lymph nodes. There, it infects B lymphocytes. The B-cell
of plague? + infection can be (1) productive, in which the virus replicates
and kills the B cell; or (2) non-productive, in which the virus
establishes a latent infection, maintained as either plasmid or
provirus (integrated into the host cell chromosome). For most
27.4 ■ Viral Diseases of the B cells, the infection is non-productive. The virus activates
Lymphatic and Blood these B cells, causing them to proliferate and produce immuno-
Vascular Systems globulins. The proliferating B cells are responsible for the large
numbers of mononuclear cells that give the disease its name.
Learning Outcomes Their numbers and appearance sometimes cause the incorrect
5. Describe the characteristics of infectious mononucleosis. diagnosis of leukemia—this is disproved when the patient spon-
6. Compare and contrast yellow fever, dengue fever, and viral
taneously recovers.
hemorrhagic fevers. A common consequence of B-cell infection is the appearance
of a specific type of antibody useful for diagnosing infectious
A number of viral illnesses affect the lymphatic system and mononucleosis; it has no pathological significance. This antibody
blood vessels. This section discusses infectious mononucleosis, type, called a heterophile antibody, will react with an antigen on
yellow fever, dengue fever, and viral hemorrhagic fevers, which the red blood cells of certain animal species, including sheep,
prominently involve the circulatory system. Other examples of horses, and oxen.
viral diseases that affect the lymphoid and circulatory systems are The proliferation of the lymphocytes causes lymph node
the human immunodeficiency virus (HIV), the cause of AIDS, and spleen enlargement. Occasionally, the spleen ruptures—
and cytomegalovirus, which are covered in chapter 28. human this is the main cause of the rare deaths caused by infectious
immunodeficiency virus disease, p. 696 cytomegalovirus disease, p. 711 mononucleosis and is most likely to occur within 3 to 4 weeks
of the start of illness.
T cells respond actively to the infection and destroy B cells
Infectious Mononucleosis with productive infections; they do this because these B cells
(“Mono” or “Kissing Disease”) display viral antigens on their surfaces. The abnormal-appearing
Infectious mononucleosis (“mono”) is a disease familiar to many lymphocytes seen in smears of the patient’s blood are effector
students because of its high incidence among young people. The cytotoxic T cells responding to the infected B cells (figure 27.8).
term mononucleosis refers to the fact that people with this disease The possibility that EBV plays a role in causing certain
have an increased number of mononuclear leukocytes in their malignant tumors (cancers) has been intensely investigated. The
blood. leukocytes, p. 339 EBV genome is detectable in almost all cases of Burkitt’s lym-
phoma and nasopharyngeal carcinoma, but evidence suggests that
Signs and Symptoms other factors are also involved in these cancers. EBV may also be
Typically, signs and symptoms of infectious mononucleosis a factor in some malignancies in patients with immunodeficiency
appear after a long incubation period, usually 30 to 60 days. They from AIDS or organ transplantation.
include fatigue, fever, a sore throat covered with pus, and enlarge-
ment of the spleen and lymph nodes. In most cases, the fever and Epidemiology
sore throat are gone in about 2 weeks and the enlarged lymph EBV is distributed worldwide. It infects individuals in
nodes in 3 weeks. Generally, people recover fully within 4 weeks, crowded, economically disadvantaged groups at an early age
although some suffer severe exhaustion and difficulty concentrat- without causing significant illness or symptoms of infectious
ing that affects them for months. mononucleosis. In more affluent populations, fewer people get
infected with EBV, so most lack immunity to it. Adolescents
Causative Agent and adults who lack antibody to the virus may get infectious
Infectious mononucleosis is caused by the Epstein-Barr virus mononucleosis when they are exposed to it later in life, such as
(EBV), named after its discoverers, M. A. Epstein and Y. M. Barr. when they enter college. Even then, in the age group of 15 to
 Part IV Infectious Diseases 681

Neutrophils Lymphocyte Monocyte

1 EBV
Epithelium
EBV attaches to
and infects epithelium
of the throat, where
it replicates and
causes pharyngitis.

Lymphatic vessel

Blood vessel

2 (a)
Virions enter the
lymphatic vessels and Abnormal lymphocyte
Lymphatic are carried to the
vessel lymph nodes. Some
virions escape the
lymph nodes and
are carried to the
bloodstream.
Lymph
node

3
Virions attach specifically
to B lymphocytes and
infect them, producing
either latent or
productive infections.

Viral genome

Latently infected B cell (b)

Productively FIGURE 27.8 Normal and Infectious Mononucleosis Blood

infected B cell Smears Pictures of (a) a normal blood smear and (b) an infectious
mononucleosis blood smear showing an abnormal lymphocyte.
? Why is this disease sometimes misdiagnosed as leukemia?
Activated
5 Productively 4
lymphocyte
infected B cell Plasma
Viral cells
genome

24 years, only about half of the EBV infections are symptom-

atic; the rest have few or no signs or symptoms.
EBV is present in the saliva for up to 18 months after infec-
tious mononucleosis and it occurs intermittently for life. Continuous
shedding of virus in saliva is common in people with AIDS or other
immunodeficiencies. Mouth-to-mouth kissing is an important mode
Effector Lysed
cytotoxic B cell
of transmission in young adults, leading to the name “kissing dis-
Heterophile antibodies
T cell ease.” The donor of the virus is usually asymptomatic and may have
Latently infected B cells are not been infected in the past without developing symptoms of infectious
attacked by T cells. They differen-
T cells respond to infection tiate into plasma cells and produce
mononucleosis. By middle age, most people have antibody to the
and destroy the lymphocytes virus, indicating past infection. There is no animal reservoir.
random immunoglobulins,
that have replicating EBV. including heterophile antibody.
Treatment and Prevention
FIGURE 27.7 Pathogenesis of Infectious Mononucleosis Antiviral medications such as acyclovir and famciclovir inhibit
? Why is infectious mononucleosis also called “kissing disease”? productive infection by the virus and are helpful in rare serious
682 Chapter 27 Blood and Lymphatic Infections

PERSPECTIVE 27.2
Walter Reed and Yellow Fever
Walter Reed was born in Virginia in 1851. Dr. James Carroll, developed classic yellow (2) an interval of about 12 days must elapse
After receiving two medical degrees, he fever. Dr. Jesse Lazear, another commission between the time the mosquito ingests the
entered the Army Medical Corps. He was member, noted that the yellow fever patient on blood of an infected person and the time it can
appointed professor of bacteriology at the which the mosquito had originally fed was in transmit the disease to an uninfected person;
Army Medical College in 1893, and in 1900, his second day of the disease and that 12 days (3) yellow fever can be transmitted from a per-
Reed was named president of the Yellow had elapsed before it bit Carroll. This timing son acutely ill with the disease to a person who
Fever Commission of the U.S. Army. At that was critical for the transmission of the dis- has never had the disease by injecting a small
time, the incidence of yellow fever was high ease because, as we now know, the mosquito amount of the ill person’s blood; (4) yellow
and its cause and transmission were unknown. can contract the infection only during a brief fever is not transmitted by soiled linens, cloth-
Reed suspected that an insect was the period when the patient is viremic. Then, the ing, or other items that have come into contact
vector of yellow fever. As early as 1881, mosquito can transmit the infection to another with infected persons; and (5) yellow fever is
Dr. Carlos Finlay of Havana, Cuba, had sug- individual only after the virus has replicated to caused by an infectious agent so small that it
gested that a mosquito might be the carrier of a high level in the mosquito. passes through a filter that excludes bacteria.
this disease, but he was unable to prove his Lazear was later working in a hospital Major William C. Gorgas, the chief sani-
hypothesis. Reed and others in his commis- yellow fever ward where he got bitten by a tary officer for Havana, used this information
sion proceeded with a series of experiments to mosquito, developed yellow fever and died. and started mosquito control measures that
try and prove mosquito transmission. They let The yellow fever commission had a mosquito- resulted in a dramatic reduction in yellow
laboratory-raised mosquitoes feed on patients proof testing facility constructed, called Camp fever cases. Later, Gorgas used mosquito
with yellow fever and then on members of Lazear in honor of their deceased colleague, to control in the Panama Canal Zone to allow
the commission. Initially, their experiment house volunteers for their experiments. construction of the Panama Canal. Previously,
seemed to have failed, because no one got ill. As a result of their studies, Dr. Reed and French workers had tried to build the canal but
However, 3 days after an experimental mos- his colleagues made the following conclusions: had failed because of heavy losses from yellow
quito bite, one member of the commission, (1) mosquitoes are the vectors of the disease; fever and malaria.

cases; however, they have no activity against the latent infection. Yellow Fever
Cortisone-like medication is sometimes useful in relieving airway
Yellow fever was first recognized with an epidemic in the Yucatan
obstruction from swollen tissue.
of Mexico in 1648, probably introduced there from Africa. One
Infectious mononucleosis can be prevented by avoiding
of the worst outbreaks of yellow fever in the twentieth century
the saliva of another person. This includes avoiding their tooth-
occurred in Ethiopia in the 1960s, causing 100,000 cases and
brushes, drinking glasses, and any other object that may be
30,000 deaths. In 1989, an epidemic of yellow fever occurred in
contaminated with saliva. There is no vaccine for this disease.
Bolivia among poor people who moved into the jungle to try to
Table 27.6 gives the main features of infectious mononucleosis.
make a living growing coca. There have been no outbreaks in the
United States since 1905, but the vector mosquito has reappeared
in the Southeast, raising the possibility of outbreaks of the disease
if the virus is again introduced (see Perspective 27.2).

Signs and Symptoms

Infectious Mononucleosis The signs and symptoms of yellow fever can range from very mild
TABLE 27.6 (“Mono” or “Kissing Disease” ) to severe. Symptoms of mild disease, the most common form, are
Signs and Fatigue, fever, sore throat, and
fever and a slight headache lasting a day or two. Patients suffer-
symptoms enlargement of lymph nodes ing severe disease, however, may experience a high fever, nausea,
Incubation period Usually 1 to 2 months
bleeding from the nose and into the skin, “black vomit” (from
gastrointestinal bleeding), and jaundice (hence, the name yellow
Causative agent Epstein-Barr virus (EBV), a DNA virus of the
fever). The mortality rate of severe yellow fever cases can reach
herpesvirus family
50% or more. The reasons for the wide variation in severity of
Pathogenesis Productive infection of epithelial cells of symptoms are unknown but are probably due to the size of the
throat and salivary ducts; latent infection of
B lymphocytes; hemorrhage from enlarged
infecting dose and the status of the host’s defenses.
spleen is a rare but serious complication.
Causative Agent
Epidemiology Spread by saliva; lifelong recurrent
shedding of virus into saliva of Yellow fever is caused by an enveloped, single-stranded, RNA
asymptomatic, latently infected arbovirus of the flavivirus family. The virus is transmitted by the
individuals. bite of infected Aedes aegypti mosquitoes, its biological vector.
arbovirus, p. 641
 Part IV Infectious Diseases 683

Pathogenesis Dengue Fever

The yellow fever virus is transmitted by the bite of an infected Dengue fever is a mosquito-borne viral disease similar to but
mosquito. It multiplies in local host cells, enters the bloodstream, milder than yellow fever. The disease is also known as “break-
and is carried to the liver and other parts of the body. Liver bone fever” because it causes severe joint and muscle pain. The
damage results in jaundice and decreased production of clotting disease is found in many tropical areas and is now endemic in
proteins. Injury to small blood vessels produces petechiae—tiny more than 100 countries worldwide. It is the most common vector-
hemorrhages—throughout the body. Viral replication—as well borne viral disease in the world—the World Health Organization
as the immune response to the infection—damages the circula- estimates that there are 50 million new cases every year and that
tory system, leading to bleeding in various tissues, and causing 2.5 billion people are at risk of contracting this disease.
disseminated intravascular coagulation (DIC). Kidney failure is a
common consequence of loss of circulating blood and low blood MicroByte
pressure. biological vector, p. 442 DIC, p. 674 Recently, 28 cases of dengue fever were reported in the Florida
Keys, a subtropical area of the United States.
Epidemiology
The reservoir of the yellow fever virus is mainly infected mos-
quitoes and primates living in the tropical jungles of Central and Signs and Symptoms
South America and in Africa. Periodically, the disease spreads Many cases of dengue fever are asymptomatic or subclinical. If
from the jungle reservoir to urban areas, where it is transmitted to signs and symptoms occur, they begin with a sudden onset of
humans by Aedes mosquitoes. fever that lasts for 2 to 7 days but may recur 12 to 24 hours later
(biphasic fever). Other signs and symptoms include headache,
Treatment and Prevention joint and muscle pain, pain behind the eyes and a maculopapular
There is no proven antiviral treatment for yellow fever. rash that may later become hemorrhagic, with development of
Yellow fever can be prevented in urban areas by spraying petechiae. In mild cases, the rash does not develop, often resulting
insecticides and eliminating the breeding sites of its main vec- in a misdiagnosis of influenza or other viral disease. Mild mucosal
tor, Aedes aegypti. In the jungle, yellow fever control is almost bleeding may occur, typically involving the nose or gums. Patients
impossible because the mosquito vectors live in the forest can- may also complain of sore throat. In some cases, abdominal pain,
opy and transmit the disease among monkeys. A highly effective vomiting, and diarrhea occur. maculopapular rash, p. 674

attenuated vaccine is available to immunize people who might Dengue fever is generally self-limiting and is rarely fatal.
become exposed, including travelers to endemic areas. The main However, a severe form of the disease—dengue hemorrhagic
features of yellow fever are presented in table 27.7. attenuated fever (DHF)—can occur in patients who experience a second den-
vaccine, p. 423 gue infection. DHF may lead to dengue shock syndrome (DSS),
which causes a weak, rapid pulse, moist skin, restlessness, and low
blood pressure. Petechiae, easy bruising and sometimes massive
gastrointestinal and vaginal bleeding occur. Later, widespread
blood clotting and DIC may develop. DHF is mostly found in
TABLE 27.7 Yellow Fever children under 15 years old and can be fatal in this population.
Signs and Often only headache and fever. Severe cases DIC, p. 674
symptoms characterized by high fever, jaundice, black
vomit, and hemorrhages into the skin. Causative Agent
Incubation period Usually 3 to 6 days Dengue fever is caused by a single-stranded RNA virus of the
Causative agent Yellow fever virus, an enveloped, single- flavivirus family. There are four closely related serotypes of the
stranded RNA virus of the flavivirus family virus (DENV1, DENV2, DENV3, and DENV4), which are trans-
Pathogenesis Virus multiplies locally at site of introduction mitted by the bite of an infected mosquito of the Aedes family.
by an infected mosquito; spreads to the The most common vector is Aedes aegypti, the same species that
liver and throughout the body by the
carries yellow fever. Another important vector of dengue viruses
bloodstream. Virus destroys liver cells,
causing jaundice and decreased production is Aedes albopictus, the Asian tiger mosquito, now spreading
of blood-clotting proteins. Hemorrhages worldwide. This vector also transmits the denguelike viral disease
and decreased strength of the heart result in chikungunya (discussed later). Both of these mosquito species feed
circulatory failure and kidney failure. only during the day.
Epidemiology Virus persists in forest primates and the
mosquitoes that feed on them, in Africa and Pathogenesis
Central and South America; human epidemics
occur when the virus infects household When an infected mosquito feeds on a human host, dengue viruses
mosquitoes that feed on humans. are injected into the bite wound, where they infect keratinocytes
Treatment No proven antiviral therapy is available. and epidermal dendritic cells. Some of these cells migrate to the
and prevention There is a highly effective attenuated viral lymph nodes, where monocytes and macrophages are recruited
vaccine. and also become infected. Dengue viruses multiply in the mac-
rophages and are then disseminated systemically in these cells.
684 Chapter 27 Blood and Lymphatic Infections

CASE PRESENTATION
The patients were two American boys, 7 along the Gulf of Mexico. Serious dengue infection or from the mother, attach to the
and 9 years old, living in Thailand, who epidemics occurred in Brazil and Cuba in infecting dengue virus strain and promote
developed irritated eyes and slightly runny 2002. its uptake by macrophages. The virus,
noses, progressing to fever, headache, and 2. The two individuals presented here had a instead of being killed, reproduces in the
severe muscle pain. It hurt them to move mild form of the disease that characteristi- macrophage. This results in the death of
their eyes, and they refused to walk because cally involves newcomers to an area where the macrophage and release of chemicals
of pain in their legs. The symptoms sub- the disease commonly occurs. In endemic that cause leaky capillaries and shock.
sided after a couple of days, only to recur areas, the disease is characterized by fever, T lymphocytes may also play a role in
at lesser intensity. No treatment was given, headache, muscle aches, rash, nausea, and this process in older children and adults.
and they were completely back to normal vomiting. In a small percentage of cases, Evidence also exists that the virus causes
within a week. Their illness was diagnosed as however, the effects of dengue infection a depression of the bone marrow, account-
dengue (pronounced DEN-gay), also known are much more serious, resulting in den- ing for the very low white blood cell and
as “breakbone fever.” gue hemorrhagic fever. This form of the platelet counts seen in dengue.
1. What causes dengue and where does it disease is characterized by bleeding and 4. Scientists at Mahidol University in Bang-
occur? leakage of fluid from the capillaries. An kok and other research centers around the
2. Why is dengue important? important result is that the blood pressure world are working on vaccines designed
drops and the blood thickens. With expert to bring dengue under control. At present,
3. What is known about the pathogenesis of
treatment, the mortality is about 1% to control efforts are largely directed at kill-
dengue?
2%. Dengue shock syndrome is another ing mosquitoes and their larvae and elimi-
4. What can be done to prevent dengue? potential development. It is character- nating water containers around houses
ized by profound shock and disseminated where the mosquitoes breed. Scientists at
Discussion intravascular coagulation (DIC) and has a Colorado State University have success-
1. Dengue is caused by any of four closely mortality above 40%. fully genetically engineered mosquitoes
related flaviviruses—DENV1, DENV2, 3. About 90% of the cases of hemorrhagic to make their cells incapable of repro-
DENV3, and DENV4. The disease occurs fever or shock occur in people who ducing dengue virus. This was done by
in large areas of the tropics and subtropics have previously been infected by a den- using another virus to introduce an anti-
around the world. In the Americas, the gue virus and have antibody to it. The sense segment of the dengue genome into
disease is transmitted mainly by Aedes remaining cases occur largely in infants the mosquito cells. This was a dramatic
aegypti mosquitoes, which are reappear- who still have transplacentally acquired accomplishment, but years of work remain
ing, after being almost eradicated in the maternal antibody against dengue viruses. before it can play a role in the control of
1960s. This vector now occurs year-round These antibodies, whether from earlier dengue.

Humoral and cell-mediated immune responses clear the viruses. macrophages die and release toxic products that cause blood clot-
The details of dengue fever pathogenesis are not well understood ting and DIC. Leukopenia (decrease in white blood cell number)
because there are no animal models for the disease. occurs. Blood pressure drops, sometimes to life-threatening levels.
The pathogenesis of DHF and DSS is better understood.
DHF and DSS generally occur in people who experience a second Epidemiology
dengue infection with a different strain of the virus from that of Dengue is an emerging disease and the fastest spreading
their first infection. Antibody-dependent enhancement (ADE) mosquito-borne viral disease in the world. It is found in mostly
of infection has been suggested to explain severe dengue disease. tropical and subtropical regions that have high rainfall and plenty
In this model, preexisting dengue antibodies from a primary den- of fresh water, conditions that aid in disease development because
gue infection recognize the viruses in the second infection and the mosquito vector breeds in standing water. These regions
bind to them, forming immune complexes. However, because include Southeast Asia, western Pacific, eastern Mediterranean,
the preformed antibodies are against a different serotype of the the Americas, and rural areas in parts of Africa and the Caribbean.
virus, they do not neutralize the viruses. Instead, they facilitate Geographical spread of the disease is linked to international travel,
viral entry into cells that express Fc receptors, especially macro- breakdown of vector control measures, and increase in mosquito-
phages, where the viruses then freely replicate. ADE thus leads to breeding habitats—such as used tires and water containers, in
increased numbers of infected cells and a high viral load. which water accumulates.
The infected macrophages produce pro-inflammatory cyto-
kines, resulting in a cytokine storm. This, along with activated Treatment and Prevention
complement and cytokines released by memory T cells, causes There is no specific treatment for dengue fever. Patients are given
changes in vascular permeability, leading to plasma leakage, that analgesics (pain relievers). They should avoid aspirin and non-
may result in respiratory distress and dehydration. Capillaries steroidal anti-inflammatory drugs such as ibuprofen because these
become fragile, resulting in the observed hemorrhages (petechiae, medications can worsen bleeding associated with dengue. Aspirin
easy bruising, and gastrointestinal and vaginal bleeding). Infected may also be associated with Reye’s syndrome. Oral rehydration
 Part IV Infectious Diseases 685

therapy is given to replace fluid lost from fever and vomiting. CHIK is not a new disease, there have been several recent large
Platelet or blood transfusions are given for DHF if bleeding and outbreaks of it, making it an emerging disease.
plasma leakage have occurred. Efforts are being made to develop The disease is caused by Chikungunya virus, an alphavirus of
antiviral medications for dengue. Reye’s syndrome, p.  535 oral the Togaviridae family. It is transmitted by Aedes mosquito spe-
rehydration therapy, p. 571 cies, mostly A. aegypti and A. albopictus. The signs and symptoms
Prevention and control of dengue and DHF have become are similar to those of dengue and include fever that typically
more important with their increasing geographic distribution lasts 2 to 5 days, followed by severe joint pain (especially in the
and incidence. Measures include controlling or eradicating the extremities—fingers, toes, wrists, and ankles) that can persist
mosquito population by destroying their breeding sites (stand- for weeks or months. Sometimes patients develop chronic joint
ing water). Environmental insecticide use is also important, problems—this persistent joint pain differentiates CHIK from
although increasing insecticide resistance is becoming a problem. dengue fever. Patients often develop a macular or petechial rash.
Biological control of the vectors (for example, using fish which Nonspecific symptoms include headache, conjunctivitis and pho-
eat the mosquito larvae), has been successful in some countries. tophobia, back pain, nausea, and general malaise. The disease is
Other prevention measures include educating people in the use of seldom fatal.
insecticide-treated mosquito bed nets and insect repellents. CHIK occurs mainly in Africa, Asia, and Southeast Asia,
Significant effort has been made in developing a dengue but recent epidemics have been reported in several other areas,
vaccine—several live attenuated vaccines are in advanced stages associated with returning travelers who inadvertently introduce
of development. Vaccine development is hampered because creat- A. albopictus to the affected regions. This vector is now present
ing a vaccine against all four viral serotypes (tetravalent vaccine) in Brazil, Central America, the United States, and many European
is difficult, and there is no animal model for the disease. The main countries, where it has adapted to favorable environmental con-
characteristics of dengue fever are presented in table 27.8. ditions. In 2007, the first European endemic outbreak of CHIK
occurred—Chikungunya virus was transmitted to A. albopictus in
Chikungunya a northeastern region of Italy by a man returning from India, lead-
ing to an epidemic that affected over 200 people.
Chikungunya, commonly known as CHIK, got its name from an
There is no specific treatment for CHIK. Analgesics and
African word meaning “that which bends up” because people with
non-steroidal anti-inflammatory drugs such as ibuprofen are used
the disease show bent posture due to severe joint pain. Although
to reduce the joint pain. Fluids are given to reduce dehydration
from fever. Chloroquine is increasingly being used successfully
TABLE 27.8 Dengue Fever to treat chronic cases of the disease. As with most mosquito-borne
diseases, CHIK can be prevented by effective vector control—
Signs and Often asymptomatic. Fever, headache, destroying the vector and protecting the population by use of
symptoms maculopapular rash and severe joint pain. In insect repellents and insecticide-impregnated mosquito netting. A
dengue hemorrhagic fever (DHF), bleeding
and shock can occur, as well as disseminated
vaccine for CHIK is currently in clinical trials.
intravascular coagulation (DIC).
Incubation period Usually 4 to 7 days Emerging Hemorrhagic Fevers
Causative agent Dengue viruses DENV1, DENV2, DENV3, and Several viral hemorrhagic fevers are considered emerging. These
DENV4—RNA viruses of the flavivirus family include Ebola and Marburg hemorrhagic fevers, among others.
Pathogenesis Dengue viruses multiply in macrophages
and are disseminated throughout the Ebola Hemorrhagic Fever
body. Infected macrophages produce pro-
Ebola hemorrhagic fever (EHF) is caused by the Ebola virus,
inflammatory cytokines, leading to leaky
blood vessels and hemorrhaging. In DHF, named after the Ebola River Valley in the Democratic Republic
antibody-dependent enhancement caused of the Congo (in Africa). There are five different species of Ebola
by reinfection is thought to enhance viral virus, four of which cause disease in humans. Ebola virus is one of
entry into macrophages, increasing viral load. two virus genera belonging to the Filoviridae family.
Macrophages die, causing blood clotting and
The Ebola virus causes severe, often fatal disease. Signs and
DIC, which may be fatal.
symptoms include rapid onset of fever, headache, abdominal pain,
Epidemiology Viruses transmitted by mosquitoes of the
joint and muscle pain, sore throat, macular rash, and bleeding. The
Aedes family, most commonly Aedes aegypti
but also A. albopictus. Found predominantly virus causes capillaries to become leaky and disrupts clotting—
in tropical and subtropical regions with patients bleed from mucous membranes and body orifices; bloody
high rainfall but increasing in geographical vomit and diarrhea are common. Bleeding is extensive and occurs
spread. DHF usually occurs in children under both internally and externally. The massive bleeding is fatal in
15 years old.
50% to 80% of cases; death is by multiorgan failure or shock.
Treatment No specific treatment. Analgesics for pain; EHF is found in various African countries. Social and eco-
and prevention oral rehydration therapy and blood or
nomic conditions often favor the development of epidemics in
platelet transfusions if bleeding occurs.
Prevention: controlling the vector population. these countries. Ebola virus has been reported in the United States
Vaccine development is underway. and England, associated with import of monkeys carrying the
virus. No fatal cases have occurred in these countries.
686 Chapter 27 Blood and Lymphatic Infections

There is no standard treatment for EHF. Patients are kept 27.5 ■ Protozoan Diseases
hydrated, and blood and O2 levels maintained. Plasma contain-
ing coagulation factors is given to control bleeding. Prevention Learning Outcomes
of EHF is difficult because the natural reservoir is unknown. It is 7. Outline the Plasmodium vivax life cycle.
suspected that fruit bats carry the virus. Ebola virus is transmit-
8. Discuss why malaria has been so difficult to control.
ted to humans from bats and other humans by contact with body
fluids. It has been found in other animal species, including gorillas
Protozoa infect the blood vascular and lymphatic systems of
and chimpanzees, but the high mortality in these infected animals
millions of people globally. Malaria is a widespread protozoan
makes it unlikely that they are reservoirs.
disease that is a leading cause of morbidity (illness) and mortal-
Once EHF occurs in a community, quick diagnosis and rapid
ity (death) worldwide. Other causative agents of cardiovascular
response by healthcare workers is essential to prevent spread to
and lymphatic protozoan diseases include Trypanosoma brucei,
other individuals. Healthcare workers treating infected people
a bloodstream parasite that causes African sleeping sickness in
should be stringent in their use of protective gear and infection-
humans, and T. cruzi, which causes Chagas’ disease (American
control measures. A vaccine is in clinical trials.
trypanosomiasis). The latter often manifests as a chronic heart
infection. Protozoans of the genus Leishmania cause visceral
Marburg Hemorrhagic Fever
leishmaniasis, with enormous splenic enlargement. There is more
Marburg hemorrhagic fever (MHF) is caused by the Marburg information available about leishmaniasis at the text website
virus, the second genus belonging to the family Filoviridae. The (www.mhhe.com/nester7). protozoa, p. 291
virus was named for the German town of Marburg when several
people died from MHF after handling infected African green mon-
Malaria
keys. The signs and symptoms of MHF are similar to those of EHF
and include fever, rash, abdominal pain, and nausea. However, Malaria is an ancient disease, as shown by early Chinese and
bleeding from the orifices is rare. Jaundice and liver failure may Hindu writings. During the fourth century b.c., the Greeks noticed
occur. Patients may be misdiagnosed because the disease symp- the association of malaria with swamps and began drainage proj-
toms resemble those of diseases such as malaria or typhoid fever, ects to control the disease. The Italians gave the disease its name,
as well as EHF. MHF has a fatality rate of up to 25%. malaria, which means “bad air,” in the seventeenth century. In
MHF is found in Africa. Cases in Europe and the United 1902, Ronald Ross received a Nobel Prize for demonstrating the
States have been associated with either handling infected animals life cycle of the protozoan cause of malaria.
or international travel. Treatment of MHF is comparable to that Malaria is the most common serious infectious disease world-
of EHF. Patients receive fluids and supportive therapy, as well as wide. In 1955, the World Health Organization (WHO) began a
plasma replacement and O2. Prevention is difficult because little is program for the worldwide elimination of the disease using insecti-
known about the natural reservoir and transmission, but it follows cides such as DDT to kill the mosquito vector, and diagnosing and
the guidelines given for EHF. Like EHF, the natural reservoir is treating infected patients. The program was initially successful—52
thought to be fruit bats. Transmission among humans is by body nations participated, and by 1960, 10 of them had eradicated malaria.
fluids and possibly aerosol droplet. Both Ebola virus and Marburg Unfortunately, the mosquito vectors began to develop resistance
virus are considered bioterrorism threats, and work is being car- to insecticides, and malaria began to reappear. In 1976, the WHO
ried out to develop vaccines against them. acknowledged that the eradication program was a failure. Today, over
300 million people are infected with the malarial parasite annually
MicroAssessment 27.4 worldwide, with more than 1 million deaths per year. More people are
dying of the disease than when the eradication programs first began.
Infectious mononucleosis occurs worldwide and is transmitted
from person to person by saliva. Yellow fever virus is transmitted
MicroByte
to humans by the mosquitoes of the Aedes genus. Dengue fever
An estimated 300 to 500 million people contract malaria each year,
(breakbone fever) is the most common vector-borne disease in
and a child dies of the disease every 40 seconds.
the world. It is also transmitted to humans by Aedes mosquitoes
and is similar to but milder than yellow fever. A severe form of the
disease, dengue hemorrhagic fever, can be fatal. Chikungunya is a Signs and Symptoms
newly emerging disease, recently reported in Europe. It resembles The first signs and symptoms of malaria are flulike, with fever,
dengue in symptoms—like dengue, it causes severe joint pain. headache, and pain in the joints and muscles. These generally begin
Ebola and Marburg hemorrhagic fevers are emerging viral diseases.
about 2 weeks after being bitten by an infected mosquito, but in
EHF is a severe, often fatal disease that causes death by massive
bleeding and multiorgan failure. MHF is similar to but less serious some cases they start many weeks later. After 2 or 3 weeks, the pat-
than EHF. tern changes and symptoms fall into three phases: (1) cold phase—
the patient feels cold and develops shaking chills that can last for as
10. What characteristic changes occur in the blood of patients with
infectious mononucleosis? much as an hour; (2) hot phase—the temperature then begins to rise
sharply, often reaching 40°C (104°F) or more; and (3) wet phase—
11. Compare Ebola, Marburg, and Chikungunya hemorrhagic fevers
in terms of symptoms, transmission, and prevention. the patient’s temperature falls, and drenching sweating occurs. This
cycle of intense symptoms—chills, fevers, and sweats—is called
12. Why does it take more than a week before a mosquito just
infected with yellow fever virus can transmit the disease? + a paroxysm. Except for fatigue, the patient feels well until 24 or
48 hours later, when the next paroxysm occurs.
 Part IV Infectious Diseases 687

Causative Agent The Plasmodium life cycle is complex, consisting of a

Human malaria is caused by protozoa of the genus Plasmodium, liver stage (exoerythrocytic stage; exo- means “outside of,”
which are transmitted by infected female mosquito species of the -erythrocytic refers to red blood cells) and a red blood cell
genus Anopheles. Five species of Plasmodium are involved— (erythrocytic) stage. 1 The infectious form of the protozoan
P. vivax, P.  falciparum, P.  malariae, P.  ovale, and P.  knowlesi injected into the human host is called a
(this last is a simian species that is more frequently being iden- sporozoite (figure  27.9). This form
tified as a cause of human malaria now that molecular is carried by the blood-
testing is available). These species differ in micro- stream to the liver,
5
6
scopic appearance and sometimes in life cycle. They
also cause various types of disease, which differ in severity Zygote
and treatment. 7 8 9

Oocyst
1

4
Blood channel
2

Liver cells

Cell nucleus

Sporozoites
Merozoites

1 Infected mosquito injects P. vivax sporozoites into a capillary

as it feeds.

3 2 Sporozoites are carried to the liver, where they multiply in

liver cells to form merozoites. The liver cells burst, releasing
merozoites into liver blood channels.

3 The merozoites infect and differentiate in red blood cells

(RBCs), becoming a ring form, then a trophozoite, then a
schizont. The infected RBC breaks open, releasing
merozoites. Some merozoites infect new RBCs, repeating
this cycle. Others infect new RBCs then differentiate in
them, forming male or female gametocytes.

4 Another feeding mosquito ingests RBCs with gametocytes.

5 The gametocytes are released as the RBCs are digested.

6 The gametocytes become gametes, and fertilization occurs,

forming a zygote.

7 The zygote becomes motile and penetrates the gut wall.

8 In the gut wall, the zygote forms an oocyst and

multiplies asexually.

FIGURE 27.9 Life Cycle of Plasmodium vivax 9 The oocyst releases sporozoites that infect the mosquito
salivary glands.
? Why do only female Anopheles mosquitoes transmit malaria?
688 Chapter 27 Blood and Lymphatic Infections

where it infects hepatocytes (liver cells). 2 There, each para- Generally, people who live in areas where malaria is endemic
site enlarges and divides asexually, producing thousands of develop immunity from repeated infections, giving them some
merozoites, which are then released into the bloodstream. Some protection from the disease. This immunity crosses the placenta
species of Plasmodium are able to form hypnozoites, which can and gives partial protection to the newborn. The greatest risk of
live in the hepatocytes for years before reproducing to form mero- death from malaria is to children over 6 months of age as this pas-
zoites. The merozoites released from liver cells infect red blood sive immunity decreases.
cells (RBCs), beginning the red blood cell stage. P.  falciparum infections are often very severe, probably
3 In an RBC, the merozoite develops first into a ring form, because this species can infect all RBCs (leading to high levels of
then into a larger motile trophozoite (feeding stage) and then a parasitemia), whereas other Plasmodium species infect only either
schizont (reproducing stage). The schizont gives rise to mero- young or old RBCs. The P. falciparum–infected cells become
zoites that are released when the RBC ruptures. The merozoites rigid and stick to each other and to the walls of capillaries. These
then enter new RBCs and multiply, repeating the cycle. tiny blood vessels become blocked, causing O2 deprivation in the
Some merozoites that enter RBCs develop into gametocytes tissues they supply. When this occurs in the brain—a condition
(specialized sexual forms), rather than becoming schizonts. called cerebral malaria—the result is particularly serious, but
Gametocytes differ from the other forms in both their appear- almost any organ can be severely affected.
ance and susceptibility to antimalarial medications. These sexual P. vivax and P. ovale malaria often relapse because they form
forms do not rupture the RBCs. They cannot develop further in hypnozoites (treatment-resistant forms) that survive in the liver in
the human host and are not important in causing the symptoms a dormant state. Months or even years later, hypnozoites can begin
of malaria. They are, however, infectious for certain species of growing in the liver, starting new erythrocytic cycles of infection
Anopheles mosquitoes and are thus ultimately responsible for after the earlier bloodstream infection has been cured.
transmitting malaria from one person to another.
4 When a mosquito feeds on an infected person’s blood, it Epidemiology
ingests the infected RBCs. 5 It digests the RBCs, releasing the Malaria was once common in both temperate and tropical areas of
gametocytes. Shortly after entering the intestine of the mosquito, the world, and endemic malaria was eliminated from the continen-
and stimulated by the drop in temperature, the male and female tal United States only in the late 1940s. Today, malaria is mostly a
gametocytes change in form to become gametes. 6 The male disease of warm climates, but even so, almost half of the world’s
gametocyte transforms into about six tiny, whiplike gametes that population lives in endemic areas. The reservoirs for malaria are
swim about until they unite with the female gamete (in much the infected mosquitoes and humans. Human-biting mosquito species
same way as the sperm and ovum unite in animals), forming a of the genus Anopheles are biological vectors of malaria, but only
zygote. the females transmit the disease because males do not feed on
7 The zygote transforms into a motile form that burrows blood. Malaria can also be transmitted by blood transfusions or
into the wall of the midgut of the mosquito and forms a cyst. among intravenous drug users who share syringes. Malaria con-
8 The cyst enlarges as the diploid nucleus undergoes meiosis, tracted in this manner is easier to treat because it involves only red
dividing asexually into numerous offspring. 9 The cyst ruptures blood cells and not the liver—only sporozoites from mosquitoes
into the body cavity of the mosquito, releasing sporozoites that can infect the liver. Some people of black African heritage are
then find their way to the mosquito’s salivary glands and saliva, genetically resistant to P. vivax malaria because their RBCs lack
from which they may be injected into a new human host. the receptors for the parasite. Also, people with certain genetically
determined blood diseases such as sickle cell anemia are partially
Pathogenesis protected against the disease. biological vector, p. 442

The characteristic feature of malaria—recurrent paroxysms fol-

lowed by feeling healthy again—results from the cycle of growth Treatment and Prevention
and release of merozoites from RBCs. Interestingly, the infec- Treatment of malaria is complicated by the fact that different
tions in all the millions of RBCs become nearly synchronous. stages in the life cycle of the parasite respond to different medica-
Thus, cell rupture and release of daughter protozoa occur at tions. Chloroquine, and chemically related mefloquine, are effec-
about the same time for all infected cells, and each release causes tive against the erythrocytic stages of sensitive strains, but will
a fever. not cure the liver infection with P. vivax or kill the gametocytes of
The spleen characteristically enlarges in malaria to cope P. falciparum. Primaquine or a newer derivative, tafenoquine, are
with the high levels of foreign material and abnormal RBCs that generally effective against the hypnozoites and the P. falciparum
it removes from the circulation. The spleen may rupture, which gametocytes. These same drugs can be used to prevent malaria.
can occur with or without trauma. The parasites also cause ane- Malaria prevention has become a global focus. In 1998, a
mia by destroying RBCs and converting the iron in hemoglobin new disease-fighting initiative called Roll Back Malaria was
to a form not readily recycled by the body. The large amount started (involving the WHO, the United Nations Children’s Fund
of foreign material in the bloodstream strongly stimulates the [UNICEF], the United Nations Development Program, and the
immune system. In some cases, the overworked immune system World Bank). The goal was to reduce malaria deaths by 75% by
fails and immunodeficiency results. 2015. The program included organizing and funding a sustained
 Part IV Infectious Diseases 689

used for many years, is no longer effective in many areas of the

TABLE 27.9 Malaria
world (figure  27.10). Resistance has also quickly developed to
Signs and Recurrent cycles of violent chills and fever newer medications such as mefloquine.
symptoms alternating with feeling healthy By 2004, it was generally agreed that for malaria to
Incubation period Varies with species; 6 to 37 days be controlled, more effective—although much more costly—
Causative agent Five species of protozoa of the genus medications are needed. Derivatives of artemisinin, the active
Plasmodium ingredient of an ancient Chinese herbal medicine, are now used
Pathogenesis Cell burst and release of protozoa causes in combination with another medication to minimize the risk of
fever; spleen enlarges in response to developing resistance. With better funding, these combinations,
removing large amount of foreign material called ACTs (artemisinin-based combination therapies), are
and many abnormal blood cells from the now widely available and are helping to reduce the incidence of
circulation; infected red blood cells stick
to each other and to walls of capillaries;
malaria. A number of potential vaccines are also being tested.
vessels block, depriving tissue of O2. The most promising vaccine so far gives only 30% protection
Epidemiology Transmitted from person to person by
to children. Indoor spraying of the insecticide DDT, the use of
bite of infected Anopheles mosquito. insecticide-impregnated bed nets, and elimination of mosquito
Some individuals genetically resistant to breeding areas are also being emphasized. Adequate funding of
infection. malaria control is a continuing problem. Table  27.9 gives the
Treatment Treatment: usually ACTs; other medicines if main features of malaria.
and prevention sensitivity known. Prevention: chloroquine
if in chloroquine-sensitive malarial The key features of the diseases covered in this chapter are high-
areas; doxycycline, mefloquine, or other
alternative if in chloroquine-resistant
lighted in the Diseases in Review 27.1 table.
areas; after leaving area, primaquine
is given for liver stage; ACTs or other MicroAssessment 27.5
medicines for resistant strains; eradication
of mosquito vectors; mosquito netting There are a number of different stages in the life cycles of the protozoa
impregnated with insecticide; vaccines that cause malaria, and they have differing microscopic appearance,
under development. antigenicity, and susceptibility to antimalarial medications. Malaria is
a leading serious, infectious disease worldwide, and sustained, well-
funded control measures are needed to control it.
13. Why is malaria contracted from a blood transfusion easier to
effort to improve access to medical care, strengthening local treat than that contracted from a mosquito?
health facilities, and supplying medications and long-lasting 14. What causes the recurrent paroxysms that characterize
insecticide-treated mosquito nets. Unfortunately, the rate of malaria?
malaria has continued to increase despite this initiative. This is 15. Are sporozoites diploid or haploid? +
mostly because chloroquine, the main antimalarial medication

FIGURE 27.10 Distribution of

Drug-Resistant Malaria No endemic malaria
Mefloquine-resistant
? Why has drug-resistant malaria Chloroquine-sensitive
developed? Chloroquine-resistant
Diseases in Review 27.1
Blood Infections

Disease Causative Agent Comment Summary Table

BACTERIAL INFECTIONS

Subacute bacterial Skin or mouth Results from organisms colonizing blood clots on abnormal Table 27.1, p. 673
endocarditis (SBE) normal microbiota heart valves, causing damage; characterized by progressing
fatigue and slight fever; can lead to a stroke.
Sepsis and septic Often E. coli, Characterized by fever, shaking chills, and rapid pulse
shock Bacteroides, or and respiration; due to widespread release of cytokines in
Pseudomonas response to systemic infection, particularly involving
species Gram-negative bacteria.
Tularemia Francisella tularensis Category A bioterrorism agent. Can be acquired in multiple Table 27.2, p. 676
(“rabbit fever”) ways, often by direct contact with an infected animal.
Characterized by an ulcer at entry site, fever, and enlarged
lymph nodes.
Brucellosis Brucella melitensis Characterized by recurrent fever, weakness, large lymph Table 27.3, p. 677
(“undulant fever”) nodes, and weight loss; acquired via contact with an
infected animal or foodborne; causative agent enters via
mucous membranes or breaks in the skin.
Plague (“black death”) Yersinia pestis Category A bioterrorism agent; spread by flea bites Table 27.5, p. 679
(bubonic plague) and aerosols (pneumonic plague);
endemic in rodent populations.

VIRAL INFECTIONS

Infectious Epstein-Barr virus Long incubation period, followed by fatigue that can Table 27.6, p. 682
mononucleosis (EBV) last for months, fever, sore throat, and enlarged lymph
(“mono”) nodes; transmitted in saliva; infects B cells, causing both
productive and latent infections.
Yellow fever Yellow fever virus Transmitted by mosquitoes; disease can be mild or severe; Table 27.7, p. 683
severe form characterized by fever, bleeding, black vomit,
and jaundice, and is often fatal. Vaccine available for
prevention.
Dengue fever Dengue virus Transmitted by mosquitoes; generally mild, although Table 27.8, p. 685
symptoms include severe joint pain; second infection can
result in dengue hemorrhagic fever (usually in children),
which may be fatal.
Chikungunya (CHIK) Chikungunya virus Transmitted by mosquitoes; characterized by fever and
severe joint pain, which may become chronic.
Hemorrhagic fevers Ebola viruses and Severe, often fatal diseases characterized by fever and
Marburg virus internal and external bleeding; spread person to person by
body fluids.

PROTOZOAN INFECTIONS

Malaria Plasmodium species Transmitted by mosquitoes; characterized by paroxysms Table 27.9, p. 689
of chills, fevers, and sweats; P. falciparum infections are
life-threatening, even in otherwise healthy people.
 Part IV Infectious Diseases 691

FUTURE CHALLENGES 27.1

Rethinking Malaria Control
Previous attempts at malaria control have relied than 20 countries still use it for malaria control, development, vaccines, biological control of the
heavily on the use of DDT, a non-biodegradable and more would use it if it were affordable. Anopheles vectors, and mosquito habitat altera-
insecticide. This chemical was banned in the Biodegradable insecticides are available, but tion, may prove more important in achieving
United States in 1972 because of its accumula- they are more expensive than DDT, and are sustained relief from the disease.
tion in the environment and its damaging effects also often toxic. The options for preventing The challenge for the future is to better
on birds such as the peregrine falcon and the malaria all seem bad, but a choice has to be understand the ecology of malaria as it applies
bald eagle. Many other concerns have been made of which ones are the least harmful, in each location, with the aim of minimizing
raised about its possible carcinogenicity and because malaria is such a problem worldwide. insecticide use and discovering new options
potential effects on human fetuses. Although Insecticides may be necessary short term, but for maintaining long-term control.
DDT is not generally used in agriculture, more other options, including education, economic

Summary
27.1 ■ Anatomy, and Physiology, and Ecology (figure 27.1) Tularemia (“Rabbit Fever” or “Deer Fly Fever”)
(table 27.2, figure 27.4)
The Heart
Tularemia is usually transmitted from wild animals to humans by
The left side of the heart receives oxygenated blood from the lungs and
exposure to the animal’s blood or via insects and ticks. The cause is the
pumps it into thick-walled arteries; the right side of the heart receives
Gram-negative aerobe Francisella tularensis, which is found through-
blood depleted of O2 from the veins and pumps it through the lungs.
out the United States except Hawaii.
Blood Vessels Brucellosis (“Undulant Fever” or “Bang’s Disease”)
Blood vessels include arteries, veins, and capillaries. Arteries have (table 27.3)
thick, muscular walls. The blood in veins is a dark color because it is Brucellosis, caused by Brucella melitensis, is usually acquired from cat-
depleted of O2. tle or other domestic animals, sometimes from wild animals. Hunters,
Lymphatics (Lymphatic Vessels) butchers, and those who drink unpasteurized milk or milk products are
Lymphatics—lymph vessels—take up fluid that leaks from capil- at increased risk for the disease. The organisms can infect via mucous
laries. They also take up bacteria, which are normally trapped by membranes and minor skin injuries.
lymph nodes distributed along the course of the lymphatics. Plague (“Black Death”) (table 27.5)
Spleen Plague is caused by Yersinia pestis, a member of the Enterobacteriaceae
with many virulence factors that interfere with phagocytosis and immunity
The spleen filters unwanted material such as bacteria and damaged
(table 27.4, figure 27.5). Bubonic plague is transmitted to humans by fleas
RBCs from the arterial blood. It becomes enlarged in diseases such as
(figure  27.6). Pneumonic plague is transmitted from person to person
infectious mononucleosis and malaria.
by aerosols. If the bacteria enter the bloodstream, septicemic plague
27.2 ■ Bacterial Diseases of the Blood Vascular System may occur. Untreated, bubonic plague mortality is 50% to 80%;
Bacteria circulating in the bloodstream can colonize the inside of the pneumonic plague mortality is almost 100%.
heart. These infections can kill a person quickly, as in acute bacterial 27.4 ■ Viral Diseases of the Lymphatic
endocarditis, or they can cause a gradual decline, as in subacute and Blood Vascular Systems
bacterial endocarditis.
Infectious Mononucleosis (“Mono” or “Kissing Disease”)
Subacute Bacterial Endocarditis (SBE) (table 27.1) (table 27.6)
Subacute bacterial endocarditis (SBE) is commonly caused by members Infectious mononucleosis is caused by the Epstein-Barr virus (EBV),
of the normal microbiota, including oral streptococci and Staphylococcus which establishes a lifelong latent infection of B lymphocytes. The 
epidermidis. Infection usually begins on structural abnormalities of the disease has a high incidence in young people and can cause exhaustion
heart. Prolonged treatment is usually required. that lasts for months (figures 27.7, 27.8).
Sepsis and Septic Shock (figure 27.3) Yellow Fever (table 27.7)
Sepsis, an infection-induced systemic inflammatory response, is com- Yellow fever is a zoonosis of mosquitoes and monkeys that exists
monly a healthcare-associated illness; many afflicted individuals have mainly in tropical jungles; it can become epidemic in humans where
serious underlying illnesses such as cancer and diabetes. If uncontrolled, a suitable Aedes mosquito vector is present. The disease involves the
it can progress to septic shock. Most fatal cases involve Gram-negative heart and blood vessels throughout the body and is characterized by
bacteria. fever, jaundice, and hemorrhaging. A highly effective attenuated vac-
cine is available for preventing the disease.
27.3 ■Bacterial Diseases of the
Lymph Nodes and Spleen Dengue Fever (table 27.8)
Enlargement of the lymph nodes and spleen is a prominent feature of Dengue fever is caused by different viruses that are transmitted by Aedes
diseases that involve the mononuclear-phagocyte system. mosquito species. The disease is similar to but milder than yellow fever
692 Chapter 27 Blood and Lymphatic Infections

and is characterized by joint pain, giving the common name “breakbone 27.5 ■ Protozoan Diseases
fever.” A more severe form of the disease, dengue hemorrhagic fever Malaria is the most widespread of the protozoan blood and lymphatic
(DHF), may occur in people reinfected with dengue and can be fatal. diseases, and is the most widespread of all serious infectious diseases.

Chikungunya Malaria (table 27.9)

Chikungunya (CHIK) is a newly emerging disease, recently reported Malaria is caused by five species of Plasmodium and is transmitted
in Europe. Like dengue, it causes severe joint pain, which may become from person to person by the bite of the females of certain species
chronic. of Anopheles mosquitoes, its biological vector. Now found mainly in
warm regions of the world, malaria survives despite massive eradication
Emerging Hemorrhagic Fevers programs. The life cycle is complex (figure 27.9); different forms of the
Ebola hemorrhagic fever (EHF) and Marburg hemorrhagic fever (MHF) organism invade different body cells and have different susceptibility to
are emerging viral diseases. EHF is a severe, often fatal disease that antimalarial medication (figure 27.10). Replication of the organism inside
causes death by massive bleeding and multiorgan failure. MHF is simi- RBCs results in the rupture of all infected cells almost simultaneously
lar to but less serious than EHF. and release of the progeny protozoa to infect new red cells.

Review Questions
Short Answer 4. Choose the one true statement about sepsis.
1. What is the significance of immune complex formation a) It is a rare healthcare-associated disease.
in SBE? b) The output of urine increases if shock develops.
2. What is disseminated intravascular coagulation (DIC)? c) It can be caused only by anaerobic bacteria.
3. What activities of humans are likely to expose them to d) An antibiotic that kills the causative organism can be depended
tularemia? on to cure the disease.
4. Why is brucellosis a threat to big-game hunters? e) Lung damage is an important cause of death.
5. Why might the Yersinia pestis from a patient with 5. Which of these statements about tularemia is false?
pneumonic plague be more dangerous than the same organism a) It can be contracted from muskrats and bobcats.
from fleas? b) Biting insects and ticks can transmit the disease.
6. Why might rodent burrows be a source of plague months after they c) The causative organism is closely related to E. coli.
are abandoned? d) A steep-walled ulcer at the site of entry of the bacteria and
7. What type of leukocytes does EBV infect? enlargement of nearby lymph nodes is characteristic.
8. Travelers to and from which areas of the world should have certifi- e) Without treatment, 9 out of 10 people can be expected
cates of yellow fever vaccination? to survive.
9. Why is a second infection with dengue virus more serious than the 6. Which of the following statements about brucellosis is false?
first? a) fevers that come and go over a long period of time gave it the
10. Which Plasmodium species causes the most dangerous form of name “undulant fever.”
malaria? b) the causative agent can infect via mucous membranes.
c) the causative agent is readily killed by phagocytes.
Multiple Choice d) the disease in cattle is characterized by chronic infection of the
mammary glands and uterus.
1. Which of the following infection fighters are found in lymph?
e) butchers are advised to wear goggles or a face shield to help
a) Leukocytes b) Antibodies protect against the disease.
c) Complement d) Interferon 7. Which statement about Yersinia pestis is false?
e) All of the above a) Growth conditions inside human phagocytes activate virulence
2. Which of the following statements about the spleen is false? genes.
a) It is located low on the right side of the abdomen. b) The bacterium can form biofilms in the flea digestive system.
b) It cleanses the blood of foreign material and damaged cells. c) Yops protein increases phagocytosis.
c) It provides an immune response to circulating pathogens. d) The organism resembles a safety pin in certain stained
d) It can help produce new blood cells. preparations.
e) It enlarges in a number of infectious diseases. e) It was responsible for the “black death” in Europe during the
3. Which one of the following statements about SBE is false? 1300s.
a) It is generally a chronic illness characterized by fatigue and 8. Which of the following statements about yellow fever is false?
slight fever. a) There is no animal reservoir.
b) It is usually caused by normal microbiota of the mouth b) The name “yellow” comes from the fact that many victims
or skin. have jaundice.
c) Infection typically occurs on the left side of the heart. c) Certain mosquitoes are biological hosts for the causative agent.
d) Injected-drug abuse can be a risk factor in developing the d) Outbreaks of the disease could occur in the United States
disease. because a suitable vector is present.
e) It can lead to a stroke. e) An attenuated vaccine is widely used to prevent the disease.
 Part IV Infectious Diseases 693

9. The malarial form infectious for mosquitoes is called a known to be endemic for the disease had come to the town 2 weeks
a) gametocyte. b) trophozoite. earlier to work and subsequently developed yellow fever. Several
c) sporozoite. d) schizont. coworkers reported getting mosquito bites while working with
him. Why is it important that the healthcare worker determine how
e) merozoite.
long it is since the workers were bitten by the mosquitoes?
10. Which of the following statements about malaria is true?
a) Transmission cannot occur in temperate climates. Critical Thinking +
b) Transmission usually occurs with the bite of a male Anopheles 1. The finding that there is an association between Chlamydophila
mosquito. pneumoniae infection and arteriosclerotic lesions raised hopes that
c) The disease is currently well controlled in tropical Africa. new methods to combat arteriosclerosis could be developed. An
d) P. falciparum infects only old RBCs and therefore causes investigator reviewing this research, however, stated that even a
milder disease than other Plasmodium species. perfect correlation between infection and lesion formation would
e) The characteristic recurrent fevers are associated with release not prove that infection causes arteriosclerosis. Moreover, even
of merozoites from RBCs. showing that therapeutic antibiotics could prevent infection and
lesion formation would not be definitive proof. Is the investigator
Applications justified in making this argument? Why or why not?
1. Some years ago, dentists and doctors began noticing an association 2. Even though genetically engineered mosquitoes might be devel-
between subacute bacterial endocarditis and prior dental work, oped that do not allow the reproduction of malaria protozoa,
and they began advising that an antibiotic be administered at the these mosquitoes would have little, if any, immediate effect on
time of dental procedures to those with known or suspected heart the spread of the disease. Why should this be so? What would
defects. What was the rationale for this advice? have to happen for these mosquitoes to significantly affect the
2. A healthcare worker in Honduras is concerned about a potential spread of malaria?
outbreak of yellow fever in his town. A laborer from a jungle area
28 HIV Disease and Complications
of Immunodeficiency
KEY TERMS
Acute Retroviral Syndrome
(ARS) Stage of HIV disease
following the incubation stage; often
manifests as flulike symptoms.
HIV Set Point Viral load in a
person with HIV disease, after the
immune system begins to respond to
the virus and viral numbers stabilize.
AIDS (Acquired Kaposi’s Sarcoma Tumor arising
Immunodeficiency Syndrome) from blood or lymphatic vessels.
End stage of HIV disease, manifest
Mycobacterium avium Complex
as severe immunodeficiency.
(MAC) Group of genetically related
AIDS-Related Complex (ARC) bacteria belonging to the genus
Group of symptoms (fever, fatigue, Mycobacterium that causes disease
diarrhea, and weight loss) that in people with AIDS.
indicate the onset of AIDS.
Viral Load Measure of severity
HAART Highly active antiretroviral of a virus infection; calculated by
therapy; cocktail of medications that estimating the concentration of virus
interfere with HIV replication. particles in involved body fluid.
HIV (Human Immunodeficiency
Virus) The causative agent of HIV
disease and AIDS.

Color-enhanced TEM of human immunodeficiency virus (HIV).

A Glimpse of History to use it to develop a blood test that showed that all AIDS patients were
infected with LAV. He sent a sample of this virus to Gallo. He also
In 1981, reports were made of an illness characterized by unusual infec- applied for a patent on his blood test.
tions, certain malignant tumors, and immunodeficiency in previously Meanwhile, Gallo’s laboratory also recovered a virus from AIDS
healthy, young, homosexual men. This illness came to be known as patients. Gallo was able to obtain large amounts of the virus, which he
AIDS (acquired immunodeficiency syndrome). By 1982, the CDC had named HTLV-III because it resembled human T-lymphotrophic viruses
convincing evidence that AIDS was caused by a new infectious agent, and discovered earlier. Using HTLV-III, Gallo perfected a blood test for
scientists around the world rushed to identify it. diagnosing AIDS and, like Montagnier, applied for a patent. These con-
To identify the cause of AIDS, the researchers relied on some impor- flicting patent claims caused a bitter scientific and legal battle that went
tant scientific advances of the 1960s and 1970s, including the discovery on for years. A settlement was negotiated in 1987 in which Gallo and
of lymphocyte subsets, the functions of T cells, and the role of cytokines. Montagnier were named codiscoverers of the test, and both the French
Another extremely important advance was the discovery of reverse and the Americans received royalties from it.
transcriptase by Doctors Howard Temin, David Baltimore, and Renato The settlement did not end the conflict. Genetic analysis showed that
Dulbecco, who received the Nobel Prize in 1975 for their work. This was Gallo’s HTLV-III and Montagnier’s LAV were actually the same virus.
followed in early 1978 by the identification of the first human retrovirus, It was eventually discovered that patient samples sent from Montagnier’s
human T-lymphotrophic virus (HTLV), in Dr. Robert Gallo’s laboratory at laboratory contained two different viruses, one of which was the LAV
the National Cancer Institute. Dr. Gallo went on to develop a method for and Gallo’s cultures had been accidently contaminated with this virus. In
cultivating the virus in lymphocytes. lymphocytes, p. 340 cytokines, 1994, the earlier settlement was modified.
p. 341 reverse transcriptase, p. 320 retroviruses, p. 320 The American share of the blood test royalties has generated many
In January 1983, an important breakthrough occurred in the labora- millions of dollars for the NIH over the years, but the value of the test
tory of Dr. Luc Montagnier at the Pasteur Institute in Paris. Using the is greater than any monetary figure. It helped prove that HTLV-III/LAV
retrovirus culture techniques developed earlier by Gallo, Montagnier caused AIDS. The virus was later renamed HIV for human immunodefi-
and his colleague Françoise Barré-Sinoussi isolated a new virus from ciency virus. It also showed that many infected people were asymptom-
a patient with lymphadenopathy syndrome, an AIDS-associated illness. atic, that they could transmit the virus, and that the epidemic was far more
They named the virus LAV, an acronym for lymphadenopathy virus. extensive than previously suspected. These findings helped generate sup-
Montagnier could obtain only a small amount of the virus but was able port for controlling the disease.

694
 Western and Eastern Europe/ East Asia
North America Central Europe Central Asia 82,000
70,000 31,000 130,000 770,000
1.5 million 820,000 1.6 million

North Africa/ South/

Middle East Southeast Asia
75,000 270,000
Caribbean
460,000 4.1 million
17,000
240,000
Latin America Sub-Saharan Africa
92,000 1.8 million Oceania
1.4 million 22.5 million 4,500
57,000
FIGURE 28.1 The Global HIV/AIDS Epidemic at the End of 2009 An estimated 2.6 million people were newly infected during the year (numbers
in red). Altogether, 33.3 million people (numbers in black) were living with HIV/AIDS. The arrows indicate an increase (↑) or decrease (↓) in the numbers
of cases as compared with 2001. ↔ Shows stable numbers over time period examined.
? What is one factor that has lead to the decrease in new HIV infection rates in many countries?

AIDS

 I
n the 20 years after HIV was identified, an enormous number of AIDS (acquired immunodeficiency syndrome) was first rec-
Americans died from AIDS. By 1994, it had become the lead- ognized in 1981 after an unusual number of Pneumocystis pneu-
ing cause of death among those 25 to 44 years old in the United monia cases were identified in previously healthy homosexual
States. In addition, nearly a million people had contracted HIV, and bisexual men in the United States. This type of pneumonia
and more than 40,000 were becoming infected every year. is caused by a fungus now called Pneumocystis jiroveci, which
Worldwide in this time period, an estimated 40 million people infects many people but is of such low virulence that it seldom
were living with HIV infection, and more people had died of AIDS causes disease in healthy individuals. In 1982, as other groups of
than died of the “black death” of Europe during the Middle Ages. people developed unusual opportunistic infections for no appar-
It became the primary cause of death in sub-Saharan Africa, and ent reason, the CDC began using the acronym AIDS to describe
spread rapidly into India and China. Billions of dollars have been the unexplained increased susceptibility indicative of underly-
spent trying to control the AIDS pandemic, with the result that ing immunosuppression. The various opportunistic infections
new HIV infections have declined in many areas (figure  28.1). became known as “AIDS-defining conditions”—diseases that
Nevertheless, AIDS is far from conquered. black death, p. 678 indicate a person has AIDS. These were useful in studying the
AIDS epidemic before its cause was known, and they still help
physicians by alerting them to the possibility that a patient has
AIDS (table 28.1). immunodeficiency disorders, p. 414
28.1 ■ Human Immunodeficiency AIDS is caused by the human immunodeficiency virus
Virus (HIV) Infection (HIV). The terms HIV and AIDS are not synonymous, and the
distinctions are very important. A person can be infected with
and AIDS HIV but may not be ill. AIDS is only the end stage of a complex
disease that has many signs and symptoms that precede immuno-
Learning Outcomes
deficiency. Three distinct terms are important to keep in mind as
1. Describe HIV structure and its replication strategy.
you read this chapter:
2. Outline the pathogenesis of HIV disease.
■ HIV infection indicates that the virus has entered the body and
3. Discuss the three main ways that HIV is transmitted.
is replicating, whether or not signs and symptoms are present.
4. Describe the treatment and prevention of HIV.
■ HIV disease indicates that the infection is causing signs and
5. Describe the prospects for an HIV vaccine.
symptoms.
695
696 Chapter 28 HIV Disease and Complications of Immunodeficiency

TABLE 28.1 “AIDS-Defining Conditions”

■ Cancer of the uterine cervix, invasive ■ Kaposi’s sarcoma
■ Candidiasis involving the esophagus, trachea, bronchi, or lungs ■ Lymphomas, such as Burkitt’s, or arising in the brain
■ Coccidioidomycosis, of tissues other than the lung ■ Mycobacterial diseases, including tuberculosis
■ Cryptococcosis, of tissues other than the lung ■ Pneumocystis (pneumonia due to Pneumocystis jiroveci)
■ Cryptosporidiosis of duration greater than 1 month ■ Pneumonias occurring repeatedly
■ Cytomegalovirus disease of the retina with vision loss or other ■ Progressive multifocal leukoencephalopathy (a brain disease
involvement outside liver, spleen, or lymph nodes caused by the JC polyomavirus)
■ Encephalopathy (brain involvement with HIV) ■ Salmonella infection of the bloodstream, recurrent
■ Herpes simplex virus causing ulcerations lasting a month or longer ■ Toxoplasmosis of the brain
or involving the esophagus, bronchi, or lungs ■ Wasting syndrome (weight loss of more than 10% due to HIV);
■ Histoplasmosis of tissues other than the lung also known as slim disease
■ Isosporiasis (a protozoan disease of the intestine) of more than
1 month’s duration

■ AIDS is the last stage of HIV disease, defined by CD4+ lym- lymph nodes, a condition known as lymphadenopathy syndrome
phocyte (helper T cell) levels and the presence of specific (LAS). Without treatment, AIDS is fatal. helper T cell, p.  356
opportunistic infections. opportunistic infections, p. 382 Epstein-Barr virus, p. 680

Causative Agent
HIV Disease In the United States, and in most other parts of the world, AIDS
Almost everyone who becomes infected with HIV develops HIV is typically caused by human immunodeficiency virus type 1
disease, which slowly destroys the person’s immune system and (HIV-1), a retrovirus belonging to the lentivirus subgroup of the
eventually ends in AIDS unless the individual is successfully retrovirus family (see Perspective 28.1).
treated with antiretroviral drugs. HIV-1 variants can be classified into subtypes based on their
genomes. Most belong to the group M (for “major”) group. Other
Signs and Symptoms groups include O (“outlier”), N (“non-M and non-O”) and P
The first signs and symptoms of HIV disease appear 6 days to (“pending identification of further human cases”). The M group,
6 weeks after initial infection and are similar to many viral infec- which is responsible for the current AIDS pandemic, is subdivided
tions. They are referred to as “flulike” and include fever, head- into genetically related clades (subtypes) designated A, B, C, D,
ache, sore throat, muscle aches, and enlarged lymph nodes. Some F, G, H, J, K, and CRFs (circulating recombinant forms—hybrids
people develop a generalized rash and central nervous system that arise during simultaneous infection with different clades). It
symptoms, characterized by moodiness and sometimes seizures is important to be able to recognize the different HIV-1 strains for
and paralysis. Signs and symptoms of this early stage of HIV dis- epidemiological studies.
ease, called the acute retroviral syndrome (ARS), coincide with
high levels of virus replication, and typically subside within 4 to
6 weeks. Some infected people either never develop ARS or do
not notice the signs and symptoms because they are so mild.
After the early stage of infection, virus levels drop and the dis-
ease enters an asymptomatic period that typically lasts for years.
This is often referred to as clinical latency. HIV continues to rep-
licate during this time period, however, so the virus still causes cell
damage and can be transmitted to others. Clinical latency ends as
the infected person becomes increasingly immunodeficient due to
declining helper T cell numbers. With severe immunodeficiency,
the individual contracts various opportunistic bacterial, viral, pro-
tozoan, and fungal infections that characterize AIDS. As one might
expect, the signs and symptoms at this stage of the disease vary
widely according to the kinds of infection. For example, a com-
mon sign is a fuzzy white patch on the tongue (hairy leukoplakia)
which is a result of latent Epstein-Barr virus (EBV) reactivation
(figure  28.2). Most people with AIDS suffer fever, weight loss,
fatigue, and diarrhea, referred to as the AIDS-related complex FIGURE 28.2 Hairy Leukoplakia
(ARC). The person may also suffer persistent enlargement of the ? What causes this AIDS-related condition?
 Part IV Infectious Diseases 697

Human immunodeficiency virus type 2 (HIV-2) is similar in RNA surrounded by a protein capsid (figure 28.3a). The viral
structure to HIV-1, but differs genetically from HIV-1 by more envelope originates from the host cell membrane and encloses
than 55%. Both HIV-1 and HIV-2 are thought to have arisen by the viral core. Various proteins make up the viral structure, and
cross-species transmission from different non-human primates. these are referred to by two different names: (1) an abbreviation
HIV-2 is the main cause of HIV disease and AIDS in parts of West of the functional role or location (for example, CA for capsid
Africa and India, although it has also appeared in the United States protein); and (2) a term that indicates the protein size in kDa (for
and other countries. HIV-2 transmission is generally less efficient example, CA is also called p24, being a protein that is 24 kDa
than that of HIV-1, and disease progression is slower. Otherwise, in size) (table 28.2).
the biology of HIV-2 is quite similar to that of HIV-1. For the Projecting from the envelope are proteins called Env, which
remainder of this chapter, we will use the term HIV to indicate have a complex structure made up of two types of subunits:
HIV-1. (1) a transmembrane glycoprotein called TM, or gp41, which
anchors Env in the viral envelope; and (2) a surface
HIV Structure HIV is an enveloped virus glycoprotein called SU, or gp120, which rests
that has two copies of single-stranded on the TM protein like a cap and is the part

Lipid Surface
envelope glycoprotein (SU)
gp120
Reverse
transcriptase (RT) Transmembrane
Integrase (IN) glycoprotein (TM)
gp41
Protease (PR)

Nucleocapsid (NC) Capsid protein (CA)

protein p24
RNA

MHC molecule from Matrix protein (MA)

host cell membrane p17

(a)

gag vif tat nef

pol vpr vpu rev env

5' LTR LTR 3'

gag is transcribed and tat rev

translated as a unit; the
protein is cleaved by viral
protease into MA, CA, NC
and p6. gag and pol are transcribed env is transcribed as a unit and the
and translated together; the protein mRNA is translated; resulting protein
(b)
is cleaved by viral protease into MA, is cleaved by cellular protease into
CA, NC, p6 and PR, RT and IN. SU and TM.

MA CA NC p6

SU TM

MA CA NC p6 PR RT IN

(c)

FIGURE 28.3 Human Immunodeficiency Virus Type 1 (HIV-1) (a) Diagrammatic representation of the virus showing important antigens.
(b) Map of the HIV-1 genome. (c) HIV-1 gene products. The gag and pol products must be cleaved by viral protease, and the env products must be
cleaved by host cell protease.
? What is the function of reverse transcriptase?
698 Chapter 28 HIV Disease and Complications of Immunodeficiency

PERSPECTIVE 28.1
Origin of AIDS-Causing Viruses
Where did the AIDS-causing viruses come species of large monkey, the sooty mangabey. sterilized Salk polio vaccine grown in simian
from? Genetic evidence indicates that the A likely hypothesis is that the AIDS-causing kidney cell cultures and there is no evidence
HIV-1 virus mutated to its present form fairly viruses “jumped” to humans from these sim- for the suggestion that the viruses resulted
recently, between 50 and 150 years ago. ian relatives, probably through contact with from biological warfare experiments by Russia
Although it first appeared in the United States their blood. Indeed, in Africa, chimps and or the United States. It is possible that AIDS-
in the 1970s, serological evidence indicates mangabeys are often killed for food, exposing causing viruses have existed for many years
that it was present in a few people in Africa humans to their blood. in people living in isolated African villages,
in the 1950s. Genetic comparisons of a large number of perhaps even for centuries. According to this
Viruses similar to HIV exist in a num- simian lentiviruses and AIDS-causing viruses idea, population increases and migration to
ber of wild and domestic animals, includ- from humans support the idea that the simian big, crowded cities allowed the viruses to
ing cats, dairy cattle, and monkeys. A virus viruses can jump to humans. In the case of spread rapidly, becoming more virulent in the
closely related to HIV-1 has been found in HIV-1, a jump to humans probably occurred process.
a single West African subspecies of chim- only once, between 1910 and 1950. There is The answer to the question about the ori-
panzees. The virus does not appear to harm no credible evidence to support some of the gins of AIDS-causing viruses might never be
the chimpanzees, suggesting that they have popular ideas on how HIV got into the human known precisely, but the question is intriguing
been living together for a long time. Another population; for example, it is unlikely that HIV and may lead to better understanding of the
virus, closely related to HIV-2, is present in a was transferred to humans by inadequately emergence of new infectious diseases.

that initially contacts a host cell. These SU proteins are important three important viral enzymes involved in HIV replication—
because they allow the virus to attach to and enter host cells. reverse transcriptase (RT), protease (PR), and integrase (IN).
The viral core has several components. Matrix protein (MA), RT, NC, and IN are tightly associated with the RNA. Table 28.2
or gp17, lies inside the envelope and has various functions during summarizes the major components of HIV.
different stages of the HIV replication cycle, including providing
structural support during virion assembly. The next layer in is the HIV Genome The HIV genome is simple yet functionally
protein capsid CA, or p24, the most abundant protein in the virus, complex. It typically has only nine genes, but there are multiple
which can be measured in the serum to detect early HIV infection. overlapping transcription options (figure 28.3b). Three genes—
The virion core contains nucleocapsid (NC) protein (p7), core gag (for “group antigen”), pol (for polymerase), and env (for
protein p6 (involved in budding of HIV from the host cell), and envelope)—code for the structural components just described.
The gag gene is transcribed and translated into a large polyprotein
that is then cleaved by viral protease to release MA (matrix protein),
CA (capsid protein), NC (nucleocapsid), and p6. Occasionally,
TABLE 28.2 HIV Components gag and pol are translated as a single unit (figure 28.3c). The
resulting protein is cleaved into MA, CA, NC, and p6 proteins, as
Protein Function well as the three viral enzymes—protease, reverse transcriptase,
Capsid protein (CA or p24) Coat protein and integrase. The env gene is translated to produce a precursor
Matrix protein (MA or p17) Stabilizes virion
protein—gp160—that is then cleaved by the host (not viral) pro-
tease into the SU/gp120 and TM/gp41 glycoproteins.
Transmembrane protein Stalklike portion of Env spikes,
(TM or gp41) anchoring Env to the viral
The six remaining genes, known as accessory genes, are all
envelope translated from spliced mRNA and code for the proteins Tat, Rev,
Surface glycoprotein Caplike portion of Env spikes,
Nef, Vif, Vpr, and Vpu. The first three of these, Tat, Rev, and
(SU or gp120) with which virus attaches to Nef, are regulatory proteins that interact with host cell proteins
host receptors to control HIV replication and release from the cell. The other
Nucleocapsid protein Attaches viral RNA to capsid proteins play key roles in host defense; for example, both Vif and
(NC; p7) Vpu counter host factors that otherwise inhibit HIV replication.
Core protein (p6) Involved in budding of HIV Vif also increases infected cell susceptibility to cytotoxic T-cell
from host cell killing, and Vpr stops host cell growth and induces apoptosis.
Reverse transcriptase (RT) Enzyme that makes copy of
viral RNA into DNA
Protease (PR) Enzyme involved in viral
Pathogenesis
protein processing The ability of HIV to cause a long-term and progressive infection
Integrase (IN) Enzyme that inserts viral cDNA is related to both its replication cycle and the cell types it targets.
into host genome Understanding key aspects of these has allowed scientists to
develop several medications that specifically target HIV.
 Part IV Infectious Diseases 699

Attachment and Entry Once HIV enters the body, the use both. The importance of CCR5 is illustrated by the observa-
SU/gp120 portion of Env attaches to CD4+ cells. Recall from tion that people who do not make normal amounts of CCR5 are
chapter 15 that helper T cells are CD4+, meaning they have CD4 less susceptible to HIV infection. chemokine, p. 341
protein on their surface. These are the main targets of HIV infec- When HIV encounters a CD4 molecule, it binds to it, induc-
tion. Monocytes, including macrophages and dendritic cells, are ing a conformational change in SU. This change exposes sites in
also CD4+ and they are thus targets of infection as well. Some the SU molecule that bind the co-receptor. SU then undergoes
studies suggest that cells that do not have CD4 can also be infected, another conformational change, leading to fusion of the host
but whether this contributes to HIV infection in the host is unclear, and viral membranes, a process facilitated by the TM (gp41).
and the mechanisms of viral entry are not well understood. Following fusion, the virus enters the host cell.
CD4, p. 369
In addition to needing CD4, the HIV attachment process Reverse Transcription and Genome Integration After entering
also requires a co-receptor, usually one of the chemokine recep- the host cell, the HIV-encoded reverse transcriptase makes a DNA
tors CXCR4 or CCR5 (figure 28.4). Most strains of HIV use copy of the viral RNA genome (figure 28.5). The RNA template
CCR5 as the co-receptor, but some use CXCR4 (particularly is degraded and a complementary DNA (cDNA) strand is made,
strains present at later stages of infection), and some strains can resulting in a double-stranded DNA copy of the original viral
RNA. This DNA is circularized and moves
into the host nucleus. Once in the nucleus,
1
the virally encoded enzyme integrase inserts
the DNA in a linear form into the host cell
chromosome as a provirus. No specific
Attachment.
Initial contact occurs nucleotide sequence in the host genome
when HIV gp120 (SU) is required for the insertion. Once a DNA
binds to the host cell copy of the HIV genome is integrated, the
CD4 receptor.
provirus is a permanent part of that cell’s
genome.
Reverse transcriptase makes frequent
mistakes as it copies the RNA genome
into DNA. Because the enzyme also lacks
Cell membrane proofreading ability, these mistakes are
left uncorrected. The overall result is that
the nucleotide sequence of the integrated
DNA copy of the HIV genome is gener-
2 gp41
Co-receptor binding.
ally slightly different from the parent mol-
gp120
Binding with a ecule. These changes allow HIV to evolve
Co-receptor co-receptor follows. This quickly, altering its antigens and thereby
CD4 (CXCR4 causes a conformational
or CCR5) change in gp120 that
avoiding the immune response.
facilitates gp41 (TM)
insertion into the host Replication and Its Consequences Once
cell membrane. the HIV DNA copy is integrated into the
host cell genome, it can be transcribed and
translated to produce new viral particles.
The developing virions bud from the host
3 cell, gaining their envelope as they do so.
Some host cells die as a result of the infec-
tion, but others survive. Infected helper
Fusion. T cells typically die—the loss of these
gp41 mediates
host-virus membrane cells is very important because of their
fusion and viral entry central role in the body’s adaptive immune
into the host cell. response. Recall that their cytokines nor-
mally regulate the action of cytotoxic
T cells, antibody production by B cells, and
chemotaxis of macrophages. In contrast,
infected macrophages usually survive (per-
haps because the level of virus replication
FIGURE 28.4 Attachment and Entry of HIV into a Host Cell Step 1: Initial contact in them is more moderate), and they release
occurs between gp120 (SU) and host cell CD4 receptor. Step 2: Attachment to a chemokine new virions over long periods of time.
receptor such as CCR5 then follows. Step 3: Membrane fusion, mediated by gp41 (TM).
These cells, along with a particular subset
? How do entry inhibitor medications work? of T cells called resting T cells, become
700 Chapter 28 HIV Disease and Complications of Immunodeficiency

8 Final viral assembly

and budding takes place.
Host cell
1
genome
Viral genome and
reverse transcriptase
enter cell.
7 Viral membrane proteins
Site of action are transported to host
of AZT and Host cell
nucleus cell membrane.
other reverse
transcriptase 2 DNA copy synthesized
inhibitors. by reverse transcriptase.

Site of
action of
RNA 6 Viral RNAs are protease
DNA translated, yielding inhibitors.
3 RNA degraded; second viral enzymes
DNA strand synthesized. (including protease)
and structural
proteins.

DNA Site of action

DNA of antiretroviral
drugs under
development.

5 With host cell activation, viral

4 DNA either circularizes (unintegrated DNA is transcribed, yielding
provirus) or integrase incorporates it messenger RNAs and viral
into host cell genome (integrated genome RNA.
provirus).

FIGURE 28.5 HIV Replication

? Which cells are the main target for HIV?

reservoirs for replicating HIV, protecting it from the rest of the such as viral RNA and unintegrated viral DNA inside the cyto-
immune system and providing a continuing source of infectious plasm of the infected cell, can also result in its death. cytotoxic
virus. Although infected macrophages do survive, they show T cells, p.  356 natural killer cells, p. 374 antibody-dependent cellular
impaired chemotaxis, phagocytosis, and antigen presentation, cytotoxicity, p. 361 apoptosis, p. 350
which further weaken the immune response.
The signs and symptoms of HIV disease and AIDS occur Disease Progression The number of HIV particles in the blood,
as the number of helper T cells declines, in turn affecting the measured by viral load (RNA copy number), changes during
function of the immune system as a whole. Destruction of helper the course of HIV disease. During the acute retroviral syndrome
T cells can occur via multiple mechanisms, including lysis fol- (ARS), the viral load rises to very high levels. This occurs as
lowing virus replication, attack by cytotoxic T cells and natural progeny virus particles are released from the billions of helper
killer cells, and formation of syncytia (fusion of infected cells T cells that become infected in the first few weeks of disease
with large numbers of healthy cells), which are then destroyed. (figure 28.6). Initially, the person does not have any detectable
Humoral antibody may play a role in this through antibody- antibodies against the virus. However, as the viral load increases,
dependent cell cytotoxicity. Apoptosis (programmed cell death) the person develops detectable amounts of HIV-specific antibod-
is also accelerated in some HIV-infected cells; this occurs by ies, the process of seroconversion. The viral load then decreases
a number of mechanisms, including induction by viral Tat and as the immune response destroys infected cells. The number of
production of certain cytokines. Accumulation of viral products helper T cells rapidly decreases as the infected cells are destroyed,
 Part IV Infectious Diseases 701

FIGURE 28.6 HIV

Acute Retroviral Asymptomatic Interval AIDS Disease Progression Levels
Syndrome of virus are very high during
the acute retroviral syndrome
Infection

Flulike symptoms Lymph node

may occur; HIV enlargement, fever, and at the end of the disease
infection Clinical latency weight loss, fatigue, Death when AIDS occurs. Antibody
spreads diarrhea, etc. tests for diagnosing the
throughout disease are often negative in
the body. Opportunistic the early stage of the disease
infections, even though infected people
malignant are highly infectious. The
tumors
disease steadily progresses in
CD4 lymphocytes the absence of symptoms, as
Blood levels

Circulating shown by the rising levels of

HIV RNA plasma viral RNA and falling
CD4+ cell count.

Circulating ? What are some of the

culturable HIV opportunistic infections
and malignant tumors
that people with AIDS
contract?

Months Years

but increases slowly again as the viral load goes down and the maintain high levels of anti-HIV antibody and HIV-specific CD8+
T cells are replenished. However, the immune response fails to cytotoxic T cells over many years. At the extreme end is a group
eliminate the virus completely, and the helper T-cell count typi- called elite controllers, who have very low viral set-point levels,
cally does not reach the preinfection level. seroconversion, p. 426 below the detection limit of conventional clinical assays.
During the clinically latent period, which can last for many
years, HIV continues to replicate in infected cells. The concentra- MicroByte
tion of virions in the bloodstream during this time remains rela- Tuberculosis is a leading cause of death among those with HIV
disease—in some countries, as many as 80% of people with
tively stable and is referred to as the HIV (viral) set point. This
HIV also have TB.
set point, which is measured as HIV RNA in plasma, is important
because it serves as a predictor of disease progression—the higher
the viral set point in a person, the more quickly they are likely to
develop AIDS. The observation that the viral set point predicts Epidemiology
disease progression has influenced HIV treatment, with a major HIV is transmitted by sexual contact, through blood and blood
focus now on keeping the viral set point as low as possible. This product transfusion, and vertically from mother to fetus or infant.
affects the infectiousness of the person as well, because HIV lev- It is not spread by insect bites or casual contact such as kissing and
els also correlate with transmission. sharing food, and there is no evidence that HIV is transmitted by
The continuous viral replication gradually reduces the helper sweat, urine, saliva (spitting), or tears, although it can be found in
T-cell population. In most cases, as seen in approximately 80% these fluids in infected people.
of HIV-infected people, the peripheral blood helper T-cell count
Sexual Contact Sex without condom protection is the primary
(normally about 1,000 cells per microliter) steadily falls at a rate
cause of HIV spread. The virus is present in semen, as well as
of roughly 50 cells per microliter per year. Even though the body
cervical and vaginal secretions. It is thought that HIV enters the
can normally replace over a billion CD4+ cells per day, the number
vaginal, rectal, penile, or urethral mucosa through tiny lesions that
of new cells made is not enough to replace the number dying.
occur during sexual intercourse. It can be transmitted by either
AIDS Signs and symptoms of AIDS usually appear when helper male-to-male or male-to-female sexual contact (female-to-female
T-cell counts fall below 200 cells per microliter. As these counts transmission does not appear to occur, or it is very rare). The risk
decrease and the immune system stops functioning properly, the for transmitting HIV through unprotected anal sex is greater than
viral load rises dramatically. The patient now begins experiencing the risk from vaginal sex, probably because the thin rectal mucosa
a variety of opportunistic infections, including malignant tumors is easily damaged. Although the risk of contracting the virus is
caused by infectious agents. greater for the receptive sexual partner, it can also be contracted
Half of untreated patients progress to AIDS within 9 to by the insertive partner. The risk of sexual HIV transmission
10 years. Rapid progressors (about 10% of infected individuals, increases in people who have sex with multiple partners or with
who have high viral set points) develop AIDS within a few years a person who has had multiple partners, in people who are the
of initial infection. At the other end of the spectrum are long-term receptive partner (especially receptive anal sex), and in people
non-progressors (about 5% to 10% of infected individuals, with who experience traumatic sex. It is also higher in those who
low viral set points), who show no decrease in CD4+ cells and have any genital irritation or lesion; other sexually transmitted
702 Chapter 28 HIV Disease and Complications of Immunodeficiency

infections can increase the risk of contracting HIV if genital ulcers in populations that breast feed. The situation is complicated in
or lesions are present, as is the case with syphilis, genital herpes, developing countries, because the antibodies in breast milk protect
and chlamydia, among others. Saliva is very unlikely to transmit the infant from other serious and widespread diseases in those
the disease, although there is some evidence that HIV can be areas. HIV-positive women in these countries are advised to breast
transmitted through oral sex. feed their infants if they do not have access to clean water, as the
use of dirty water for formula feeding leads to significant infant
Blood and Blood Products The next most important mode of
morbidity and mortality.
transmission of HIV is through blood and blood products. The
virus can be transmitted through whole blood, concentrated red or Treatment and Prevention
white blood cells, concentrated clotting factors, or plasma. Before
Medications used to treat HIV infections are called antiretrovirals
tests were available for screening blood products, HIV-infected
(ARVs). The most commonly used ARVs are reverse transcriptase
people unknowingly transmitted the virus to thousands of transfu-
inhibitors and protease inhibitors, but there are other classes of
sion recipients, including hemophiliacs who receive transfused
HIV ARVs, including fusion inhibitors, integrase inhibitors, and
clotting factor VIII to treat bleeding episodes. In 1985, an HIV
entry inhibitors. Reverse transcriptase inhibitors include nucleo-
screening test became available, and today blood products are
side reverse transcriptase inhibitors (NRTIs) and non-nucleoside
routinely screened for antibodies to HIV-1 and HIV-2 (by ELISA)
reverse transcriptase inhibitors (NNRTIs). These categories were
and for p24 (the capsid protein) or for HIV RNA. Blood donors
described in chapter 20 and are summarized in table 28.3. The
are also screened for self-reported risk factors. This approach has
mechanism of action of one common NRTI is illustrated in
significantly reduced the risk of transmission by blood products
figure 28.7. NRTIs, p. 476 NNRTIs, p. 476 protease inhibitors, p. 476
and organ transplants. ELISA, p. 432
integrase inhibitors, p. 476 entry inhibitors, p. 474
People who abuse injected drugs often share needles, so HIV
Antiretrovirals are usually given in combinations, an approach
transmission by blood is still a major factor in the disease pan-
referred to as highly active antiretroviral therapy (HAART).
demic. Whenever two people share a hypodermic needle, there
HAART is effective because each medication contributes to sup-
is a risk of spreading blood-borne pathogens such as HIV and
pressing the virus. Also, when the drugs are used in combination,
hepatitis B. People who get tattoos and body piercings are also at
resistant mutants are less likely to develop, despite the high muta-
risk from this route of transmission, unless universal precautions
tion rate of HIV. This is because it is unlikely that any one virion
are followed (using sterile techniques, new needles, new inks, and
will simultaneously develop resistance to all the medications at
so on). HIV can be transmitted to healthcare providers by a stick
the same time, especially when replication levels are suppressed.
from a needle used on a HIV-positive person, but the risk is very
Research indicates that early (rather than late) treatment of HIV
low—0.3% (compared with a risk of nearly 30% with a virus such
disease can significantly reduce the risk of death from it.
as hepatitis B).
Most current HAART treatments consist of three drugs: two
Vertical Transmission The third most important mode of HIV NRTIs and either a protease inhibitor or an NNRTI. Efforts are
spread is from mother to infant. HIV can be transmitted to the being made to simplify the treatment by developing fixed-dose
fetus in utero via the placenta, at childbirth, or by breast feeding. combinations that contain more than one medication. By reduc-
Without antiretroviral treatment of the mother, approximately ing the number of pills a person needs to take, patient compliance
one-third of newborns will contract HIV. However, if the mother increases. This, in turn, helps prevent resistance from developing
does not breast feed and receives antiretroviral treatment, and the and makes the treatment more effective.
infant is delivered by cesarean section—all common practices HAART does not cure HIV disease. It is used to improve the
among HIV-positive women in the United States—this risk drops quality of the person’s life, reduce the risk of other complicating
considerably (to about 1%). diseases, and lower the HIV set point. The latter is particularly
Breast feeding carries a significant risk of mother-infant important because lowering the number of replicating viral par-
transmission, contributing about 15% to 20% of infant infections ticles decreases the chance that one of them will mutate to drug

TABLE 28.3 HAART Medications

Medication Actions Examples
Entry inhibitors Interfere with viral entry into host cell Fuzeon, maraviroc
Integrase inhibitors Prevent integration of HIV cDNA into host genome Raltegravir
Maturation inhibitors Inhibit gag gene processing and virion production Bevirimat (in clinical trials)
NRTIs (nucleoside reverse Chain terminators that stop DNA replication AZT, D4T, 3TC
transcriptase inhibitors)
NNRTIs (non-nucleoside reverse Inhibit reverse transcriptase Nevirapine, delavirdine
transcriptase inhibitors)
Protease inhibitors Prevent virions from maturing Saquinavir
 Part IV Infectious Diseases 703

Normal painful peripheral nerve injury; inflammation of the pancreas;

thymine rash, mouth, and esophagus ulceration; and fever. Some of the
nucleotide AZT
protease inhibitors cause kidney stones, nausea, diarrhea, and
O O sometimes diabetes.
O O
Another major factor limiting the use of anti-HIV medica-
P O P O
tions is their cost, which can make anti-HIV medications unaf-
O O fordable in many parts of the world. Negotiated price decreases,
CH2 O
T
CH2 O
T
the approval of three generic drugs by the U.S. Food and Drug
Administration in 2005, and the provision of drugs by the U.S.
government to several highly affected countries, have made treat-
OH N3 ment available to many more HIV patients worldwide.

AZT is incorporated Preventing Infection There is no approved vaccine that prevents

into DNA chain HIV infection, although clinical trials continue. Therefore, pre-
during replication. vention efforts must be aimed at avoiding transmission.
Many people with HIV disease do not know they are infected,
and therefore all blood and blood products should be considered
as potentially containing the virus. Infectious HIV persists in
samples of blood plasma for at least a week after they are taken
from AIDS patients. HIV on objects and surfaces contaminated
O O by body fluids is easily inactivated by high-level disinfectants and
O P O O P O heat at 56°C or more for 30 minutes. Freshly opened household
sodium hypochlorite 5.25% bleach, diluted 1:10, is a cheap and
O O
effective disinfectant for general use. high-level disinfectant, p. 116
The normal nucleotide CH2 O CH2 O
in (a) ends in 3'OH, to T T Educating people about HIV and how it is transmitted is a
which another powerful weapon against the AIDS epidemic. The virus is not
nucleotide can be highly contagious, and the risk of contracting and spreading it
added for elongation O N3
of the DNA strand. can be eliminated or significantly reduced by avoiding activities
The AZT in (b) ends O P O that might transmit it (table 28.4). People who are unsure of their
in 3'N3, preventing HIV status, and especially those at increased risk of contracting
elongation. O
CH2 O
A

OH Behaviors That Help Control

TABLE 28.4
the AIDS Epidemic
(a) (b)
FIGURE 28.7 Mode of Action of Zidovudine (AZT) 1. Abstinence.
(a) Normal elongation process of DNA in which reverse transcriptase 2. Staying in a mutually monogamous relationship.
catalyzes the reaction between the OH group on the chain with
3. Avoiding sex with people at risk for HIV infection (see
the phosphate group of the nucleotide being added. (b) Reverse
table 28.5).
transcriptase catalyzes the reaction of AZT with the growing DNA
chain. Since AZT lacks the reactive OH group, no further additions to 4. Avoiding genital and rectal trauma. Small breaks in the skin
the chain can occur, and DNA synthesis is stopped. AZT is a and mucous membranes allow HIV to infect.
nucleoside. It becomes phosphorylated after entry into the cell. 5. Avoiding sex when sores from herpes simplex or other causes
? Why is AZT given to HIV-positive pregnant women and to are present. These are sites where HIV can infect.
their newborns? 6. Not engaging in unprotected anal intercourse. Receptive anal
intercourse carries a high risk of HIV transmission.
7. Using latex condoms from beginning to end of sex.
resistance. It is important to remember, however, that HAART Polyurethane condoms are a reasonable alternative for those
does not eliminate HIV provirus hidden in host cell genomes. If allergic to latex. Condoms made from other materials are not
medications are stopped, the viral set point rebounds to pretreat- reliable for disease prevention, nor are those marketed in
many countries outside the United States. Oil-based lubricants
ment levels. can not be used with latex condoms. Condoms for women are
Although HAART can be very successful in lowering the available.
viral set point, there are problems associated with this treat- 8. Postponing pregnancy indefinitely if you are a woman infected
ment. Many HIV strains are now resistant to the medications, with HIV. If you are not sure of your HIV status, have blood
and these are spreading. The medications can also have several tests to rule out HIV disease before considering pregnancy.
toxic effects. For example, zidovudine (azidothymidine, or AZT) 9. Using extreme care to avoid needles, razors, toothbrushes, etc.,
can cause anemia, low white blood cell count, vomiting, fatigue, that could be contaminated with someone else’s blood.
headache, and muscle and liver damage. Other NRTIs cause
704 Chapter 28 HIV Disease and Complications of Immunodeficiency

HIV transmission from infected mother to newborn can be

People at Increased Risk
TABLE 28.5 prevented in two-thirds of cases by giving AZT to the mother dur-
for HIV Disease
ing pregnancy and to the newborn infant for 12 weeks, or by pro-
1. Injected-drug abusers who have shared needles. viding nevirapine (NVP) to the mother at delivery and the infant
2. People who received blood transfusions or blood products within 3 days of birth. Newer, more potent combinations of AZT
between 1978 and 1985. and other antiretroviral therapies are widely used and may be more
3. Sexually promiscuous men and women, especially sex workers, effective, but their long-term safety is still being evaluated. Recent
drug abusers, and men who have sex with men. studies suggest that the use of ARVs during the breast feeding
4. People with history of hepatitis B, syphilis, gonorrhea, or
period, in the mother or the infant, or both, can reduce transmis-
other sexually transmitted infections that may be markers for sion by this route. Elective cesarean section significantly reduces
unprotected sex with multiple partners. the risk of HIV transmission to the newborn baby.
5. People who have had blood or sexual exposure to any of the Better treatment and prevention of opportunistic infections
people listed. and improved antiviral therapy against HIV have significantly
lengthened the asymptomatic stage of the disease and prolonged
life once AIDS develops (figure 28.8). The main features of HIV
disease are presented in table 28.6.
the virus, should get tested—HIV-positive individuals who know
their status can ensure that they do not spread the virus to others; HIV Vaccine Prospects
they can also receive antiretroviral treatment that lowers the viral There are no approved vaccines for preventing HIV disease,
set point and helps reduce transmissibility (table 28.5). although development of potential vaccines began soon after
As mentioned earlier, most HIV transmission occurs the discovery of HIV. A vaccine might be used in either of two
between people having unprotected sex. Use of latex condoms ways—preventive or therapeutic. A preventive vaccine would
during sex significantly reduces HIV transmission. Both male immunize uninfected individuals against the disease by inducing
and female condoms are available, although male condoms are antibodies that prevent infection. A therapeutic vaccine would
less expensive and more readily available. Studies also show boost the immunity of those already infected with HIV before they
that circumcision lowers the risk of HIV transmission to hetero- become severely immunodeficient.
sexual men by up to 60%. Several countries that have high HIV
infection rates are engaged in developing programs promoting Vaccine Challenges
circumcision as a preventive measure. Condom use is still a
HIV vaccine development has been hampered by several major
cheaper way of preventing spread of the disease, however, pro-
problems. First, HIV is genetically very variable, due to the high
viding that reliable condoms are available. Further, there is some
error rate when reverse transcriptase copies the viral genome into
concern that circumcised men may not realize that they are still
DNA. This antigenic variability of HIV enormously complicates
at risk of HIV or other sexually transmitted infections if they
the task of developing an effective vaccine against the virus—a
have unprotected sex.
vaccine that is effective against one antigenic variant may not be
An approach called pre-exposure prophylaxis, or PrEP, has
effective against another.
recently shown great promise in preventing the spread of HIV.
In one study, women using a newly developed vaginal microbi-
cide gel that included an active ARV to block HIV infection had
significantly reduced HIV infection rates. Another recent study
showed that providing two oral ARVs to high-risk, uninfected
60,000
individuals protected them against HIV infection, reducing their
risk of contracting the virus by almost half. 50,000
Since 1985, transmission by blood and blood products has
40,000
been lowered dramatically by screening potential donors for
Deaths

HIV risk factors and testing their blood for HIV and antibodies 30,000
to HIV. The risk is now estimated to be less than 1 in 1.5 mil-
lion transfusions. Screening tests have also markedly reduced the 20,000

risk of HIV transmission from artificial insemination and organ 10,000

transplantation. Also, medications are effective for preventing
AIDS acquired from HIV-contaminated instruments. Hundreds 0
1995 1997 1999 2001 2003 2005 2009
of needle and syringe exchange programs help prevent the spread Year
of HIV and other blood-borne diseases among injected-drug
abusers. In these programs, people are given sterile syringes and FIGURE 28.8 Deaths Due to AIDS, United States, 1995–2009
needles in exchange for used ones. Programs also provide drug Between 17,000 and 18,000 people still die from AIDS every year
rehabilitation efforts and education about condom use and other in this country.
safer sex practices. ? What caused the marked drop in HIV-related deaths after 1995?
 Part IV Infectious Diseases 705

A third challenge to HIV vaccine development is the fact that

TABLE 28.6 HIV Disease
HIV can avoid humoral and cell-mediated immune responses. It
Signs and Over half develop fever, sore throat, head does this by becoming latent (persisting as a provirus), and also
symptoms and muscle aches, rash, and enlarged by causing the formation of syncytia. The syncytia allow virus
lymph nodes early in the infection. After
particles to pass from cell to cell without contacting antibodies,
an asymptomatic period, symptoms result
from immunodeficiency and include cytotoxic T cells, and other immune components carried by the
unusual malignant tumors, pneumonia, bloodstream. syncytia, p. 396
meningoencephalitis, diarrhea, etc. Further complicating matters is the fact that infected individu-
Incubation Usually 6 days to 6 weeks for acute als do indeed have an immune response against the virus, but it
period symptoms; immunodeficiency symptoms is not sufficient to control the infection. Nonetheless, a vaccine
within 10 years in half the infections (10% is considered the most important way to control the HIV/AIDS
within 5 years and 90% within 17 years). pandemic.
Causative agent Human immunodeficiency virus type 1
(HIV-1), many subtypes and strains. HIV-2 Vaccine Research Because HIV does not cause AIDS in species
mainly in West Africa. other than humans, vaccine research relies heavily on the use of
Pathogenesis HIV infects various body cells, notably a model system involving related viruses in non-human primates.
those vital to specific immunity, CD4+ This includes SIV (simian immunodeficiency virus) as well as
T lymphocytes and antigen-presenting cells. chimeric viruses between HIV and SIV called SHIVs. One of the
T cells killed, numbers slowly decline until
approaches to producing an effective vaccine was to develop an
the immune system can no longer resist
infections or development of tumors. attenuated HIV strain. Unfortunately, although effective in non-
human primates, this vaccine was shown to be too risky for use
Epidemiology Three main routes of transmission of HIV:
intimate sexual contact, via transfer of blood
because HIV’s high mutation rate allows it to revert to a patho-
or blood products, and from mother to child genic state.
around the time of childbirth. Transmission There are several new approaches to vaccine development
can also occur by breast milk, and possibly currently underway. Candidate vaccines undergo an extensive
by oral-genital contact. evaluation for safety and immunogenicity in experimental ani-
Treatment Treatment: HAART therapy, consisting mals before they are tested in humans. At least 10 experimental
and prevention of combinations of several anti-HIV vaccines have been developed and tested in humans, with mostly
medications, effective for many AIDS
negative results. Vaccine research efforts that have been or are
patients, and delays progression of HIV
disease to AIDS. Not a cure, and too being evaluated include:
expensive for most of the world’s HIV ■ Recombinant vector vaccines. These are viruses that have
disease patients. No vaccine yet available.
Medications and vaccines can prevent many been engineered to carry HIV genes; for example, vaccinia
of the infections that can complicate HIV virus containing HIV envelope genes. Recently, a vaccine
disease. Anti-HIV medications and cesarean using replication-deficient adenovirus containing synthetic
section decrease mother-to-newborn HIV-1 gag, pol, and nef genes went to clinical trials (called
transmission. Effective preventive measures:
the STEP study). The trial was stopped in 2007 because it was
sex education of schoolchildren, needle
exchange programs for drug addicts, found that the vaccine did not protect recipients from acquir-
use of condoms. ing HIV, and more importantly, in some cases it actually
increased the person’s risk of becoming infected. Research
is being continued with adenovirus serotypes that are safer
for human use. A more promising trial, the RV144 trial in
Thailand, tested a canarypox virus also containing HIV genes
The lack of a good animal model further challenges vac-
env, gag, and pol. poxviruses, p. 308
cine development. Until recently, the only species that could be
infected with HIV were the great apes (such as chimpanzees); ■ Recombinant HIV-1 envelope glycoprotein vaccines.
although these animals generally do not develop AIDS when These are made by splicing genes for Env glycoproteins into
infected with HIV, their antibody response can be monitored. tumor cell lines so that these cells produce large quantities of
Development of a vaccine would require a better understanding of the glycoproteins that can be used as a vaccine (in a similar
the immunopathogenesis of HIV disease, which an animal model way to which the hepatitis B vaccine is made). However,
could provide. Different chimeric mouse models that can be used these vaccines protect only against the exact antigenic variant
to study HIV infection and pathogenesis have been developed. used to make them, and they do not cause a cytotoxic T-cell
These mice have been “humanized,” meaning that a variety of rel- response.
evant human cell types (such as helper T cells and macrophages) ■ DNA vaccines. Plasmid DNA engineered to encode HIV-1
have been introduced into them. They can be infected with HIV antigens that will be expressed in tissues is a promising
and they develop some symptoms of HIV disease, thus showing vaccine strategy. However, this approach requires multiple
potential as tools for clinical evaluations of HIV disease and to injections of high doses of DNA vaccine for an effective
assess candidate HIV vaccines. immune response. Research is focused on improving these
706 Chapter 28 HIV Disease and Complications of Immunodeficiency

CASE PRESENTATION 28.1

The patient was a young woman from West 4. Could her baby and the baby’s father have peptides specific to group O strains, and
Africa showing generalized enlargement of the disease? the virus was isolated from her blood.
her lymph nodes. She had recently moved to 5. Does this case suggest that any changes Nucleic acid sequences of the env, gag,
the United States. She had no history of drug should be made in the way HIV disease is and pol genes matched those of
abuse or of receiving blood transfusions. She diagnosed? previously isolated group O strains.
had had three sex partners in her life. Her Group O strains of HIV-1 were first found
single pregnancy the year before her arrival Discussion in Cameroon, a country on the West
was delivered by emergency cesarean section. 1. This patient could well have had HIV African coast, where they account for 6%
She and the baby’s father had routine tests for disease/AIDS because of her persistent, of HIV infections.
HIV at that time, and both tests were negative. generalized lymphadenopathy and very 4. Despite the absence of symptoms, both
The baby and the father remained well. Ten low CD4+ cell count. Her HIV strain her baby and the baby’s father could have
years before her HIV evaluation, the woman might have differed enough from the the disease. The baby’s risk of infection
had been treated for a fever by scarification pandemic group M HIV-1 strains, so that was reduced but not eliminated by the
(deliberately making superficial cuts in the the usual laboratory tests did not detect emergency cesarean section, and infection
skin) performed by a native healer; the scarifi- antibody to it. could have occurred subsequently if it
cation was done with a razor blade, the sterility were breast-fed. The mother’s very low
2. The woman could have contracted
of which is unknown to the patient, and had CD4+ cell count suggests a high level of
an AIDS-causing virus during sexual
been repeated 4 years before her evaluation. viremia and, therefore, increased risk of
intercourse, or from a blood-contaminated
Her initial test results included non- transmitting the disease. She could have
razor blade used for scarification.
diagnostic lymph node biopsies and a negative infected the baby’s father during sexual
Contracting such a virus from unsterile
HIV test (ELISA). A repeat HIV test some intercourse, or he could have infected her.
instruments during her emergency
months later was weakly positive by ELISA, 5. This patient was the first case of
cesarean is a possibility, but this seems
and the confirmatory Western blot showed group O HIV disease identified in the
less likely because of the short time
only questionable reactions of the patient’s United States. Studies indicate that
span before presenting with severe
serum with gp41 and two other HIV antigens. previous introductions, if any, were not
immunodeficiency.
A test for HIV-2 was negative. A CD4+ T lym- accompanied by spread of the virus.
phocyte cell count was very low. 3. The CDC has a global surveillance system
designed to detect cases like this because Nevertheless, federal agencies worked
1. Could this patient have HIV disease? they raise the possibility of new or rare with manufacturers of HIV tests to
If so, how could the negative tests be AIDS-causing viruses being introduced increase sensitivity of the tests to group O
explained? into a population. In the present case, strains.
2. Does the history give any possible ways samples of the patient’s blood were
in which she could have contracted HIV examined by the CDC and she was shown Source: Centers for Disease Control and Prevention. 1996.
disease? to be infected with a group O HIV-1 Morbidity and Mortality Weekly Report 45(6): 122.
3. How could the diagnosis be established? virus. The patient’s serum reacted with

vaccines. Currently a new approach, known as prime-boost, being repeatedly exposed to HIV. Potent and broadly effective
is being evaluated—in this method, a DNA vaccine primes antibodies produced against HIV by some of these people are now
the body for a response, and a second vaccine then boosts an important focus of vaccine researchers looking to identify bet-
this response. One candidate uses a DNA vaccine encoding ter antigens for vaccines.
HIV Gag protein, followed by recombinant vector vaccine.
DNA vaccines, p. 425 MicroAssessment 28.1
■ Peptide vaccines. The potential of synthetic peptides is also The signs and symptoms of people with AIDS are mainly due to the
being explored. The amino acid sequence of the surface opportunistic infections and tumors that complicate HIV disease.
proteins of HIV is known; thus, peptides that exactly mimic HIV is not highly contagious, and the AIDS pandemic could be
immunologically important segments of these proteins can stopped by changes in human behavior. Highly active antiretroviral
be synthesized and injected to induce an immune response. therapy (HAART) has given many patients with AIDS miraculous
peptide vaccines, p. 425
improvement.
1. What is the difference between HIV, HIV disease, and AIDS?
Despite the difficulties of developing an effective HIV
2. Why is it important for epidemiologists to be able to identify
vaccine, a vaccine continues to be the best hope for eventu- different HIV-1 strains?
ally controlling the HIV/AIDS pandemic. Findings that have
3. If AIDS was present in Africa in the 1950s, why did it not appear
encouraged vaccine researchers include the observation that in the United States until the 1970s? +
some sex-workers in Africa have not become infected, despite
 Part IV Infectious Diseases 707

COMPLICATIONS OF ACQUIRED IMMUNODEFICIENCIES

28.2 ■ Malignant Tumors (epidemic KS) is associated with HIV disease, and its incidence
rose dramatically with the onset of HIV disease and AIDS. It was
Learning Outcomes so common among early AIDS patients that it became an AIDS-
6. Explain how HIV disease might increase the risk of developing
defining condition, even though signs and symptoms usually
malignant tumors. appear before the person becomes severely immunodeficient.
7. Outline the relationship between Kaposi’s sarcoma and KSHV
AIDS-associated KS is a more aggressive tumor than classic KS
(human herpesvirus-8). and can involve the organs, often becoming fatal. The fourth form
of KS (iatrogenic or transplant-related KS) occurs in organ trans-
Certain malignant (cancerous) tumors are associated with HIV plant patients who receive immunosuppressant medications.
disease and other acquired immunodeficiencies. Most of these In 1994, a previously unknown herpesvirus, human
malignancies fall into one of three types: Kaposi’s sarcoma, herpesvirus-8 (HHV-8), was identified in Kaposi’s sarcomas.
lymphomas, and carcinomas in the anal or cervical epithelium. The virus can be detected in all cases of KS, whether associ-
They tend to metastasize (meaning spread to different areas of the ated with immunodeficiency or not, and it is also referred to as
body) and are difficult to treat. Evidence indicates that viruses are Kaposi’s sarcoma–associated herpesvirus (KSHV). KSHV is
involved in their development—it is thought that certain viral anti- also associated with other malignant tumors, including multiple
gens, along with cytokines, cause a host cell to multiply rapidly. myeloma, and can be detected in the saliva of patients with HIV
These cells then mutate (possibly the result of insertion of viral disease. It is present in about 25% of healthy U.S. adults.
DNA into their genome) and become malignant. The malignant In Kaposi’s sarcoma, KSHV infects the endothelial cells of
cell escapes detection and destruction by immune surveillance blood and lymphatic vessels and persists in the nucleus as a latent
because cell-mediated immunity is defective, so the cell multiplies episome (not integrated). Although no new virus particles are
unchecked. viruses and human tumors, p. 323 made, the infected cells express a few virally encoded proteins.
One of these, latency-associated nuclear antigen-1 (LANA-1),
suppresses genes needed for full viral production. Another causes
Kaposi’s Sarcoma the infected cells to become spindle-shaped. A few other viral
Kaposi’s sarcoma is an unusual tumor arising from blood or proteins are also made. These cause the host cell to release pro-
lymphatic vessels. It is characterized by papular nodules that are inflammatory cytokines that induce inflammation, and growth
red, brown, purple, or black in color (figure  28.9). The nodules factors that stimulate cell proliferation and survival, and lead
are typically found on the skin but can develop in the mouth and to formation of new blood vessels (angiogenesis). Although
respiratory and gastrointestinal tracts. typically latent, KSHV can become lytic, killing the infected cell.
Kaposi’s sarcoma (KS) occurs in four epidemiological forms. Unexpectedly, cytokines released during the lytic cycle also pro-
It was first described as a rare disease in older men of Mediter- mote tumor formation and angiogenesis.
ranean and Eastern European origin—this became known as clas- KSHV does not always cause Kaposi’s sarcoma—in
sic KS. Later, it was described among all age groups in certain immunocompetent people, infection with this virus is typically
regions of sub-Saharan Africa (endemic KS), where it is now the asymptomatic. However, almost 50% of people with HIV infec-
most commonly occurring cancer. Neither classic nor endemic tion develop KS after acquiring KSHV, indicating that HIV sig-
KS is associated with immunodeficiency. The third form of KS nificantly increases the risk of developing this malignancy. This
suggests that HIV itself may play a part in developing KS, and
research has indicated that the HIV Tat protein is important in
activating the lytic life cycle of KSHV. Other studies have started
to examine whether host genetic variants, environment, and routes
of infection also play a role.

MicroByte
Kaposi’s sarcoma was originally described by Hungarian
dermatologist Moritz Kaposi in 1872, but it became an AIDS-
defining illness in the 1980s.

B Lymphocyte Tumors
Lymphomas are a group of malignant tumors that arise
from lymphocytes. Most lymphomas affect B cells, although
T-cell lymphomas also occur. AIDS patients get B-cell lympho-
mas at a significantly higher rate than that of the general pub-
FIGURE 28.9 Kaposi’s Sarcoma Lesions lic. Epstein-Barr virus (EBV) plays a role in some types of the
? What virus is detected in all cases of KS? AIDS-associated B-cell lymphomas. This virus probably plays an
708 Chapter 28 HIV Disease and Complications of Immunodeficiency

indirect role in lymphoma formation rather than being the direct Examples of Immunizations
cause—HIV infection causes activation of latent EBV infection, TABLE 28.7 for Immunocompetent People
with release of the virus to infect new B cells. This then causes with HIV Disease
B-cell proliferation (leading to lymph node enlargement).
Malignant B-cell clones are thought to arise from this population Recommended for
of rapidly dividing cells. Epstein-Barr virus, p. 680 Recommended for All High-Risk People
EBV-associated lymphomas include cerebral (brain) lym-
Hepatitis B Hepatitis A
phoma and immunodeficiency-associated Burkitt’s lymphoma—
both types are significantly more common among AIDS patients Influenza Haemophilus influenzae type b
than in the general public. Burkitt’s lymphoma also occurs in two Pneumococcal polysaccharide Human papillomavirus (HPV)
other forms: endemic and sporadic. The endemic variant is found Diphtheria, tetanus, pertussis Meningococcal
in Africa, primarily in children who have chronic malaria, which (DTaP)
is thought to reduce their resistance to EBV. Sporadic Burkitt’s Measles-mumps-rubella (MMR)*
lymphoma resembles the other types and is also thought to be
Varicella zoster*
associated with an impaired immune system.
*Only recommended if the person has a CD4+ count above 200 μl/ml.
Cervical and Anal Carcinoma
Carcinomas (cancers) of the cervix and anus are strongly associ- people with HIV disease as early in the disease process as possible
ated with human papillomaviruses (HPVs) types 16 and 18. The (table 28.7). This section presents some examples of infectious
cells involved in these cancers are epithelial cells and therefore diseases common in patients with immunodeficiency.
differ from those in Kaposi’s sarcomas and lymphomas. HPV is
transmitted during sexual activity and infects the cervical and anal Pneumocystis pneumonia (PCP)
epithelium, appearing to cause increased replication of the cells
by blocking expression of a cellular gene responsible for control- Pneumocystis pneumonia (PCP), a severe, infectious lung dis-
ling cell growth. HPV replication increases as the host’s cellular ease, was recognized just after World War II in Europe when it
immunity declines with AIDS, organ transplantation, or other killed malnourished, premature infants in hospitals. Subsequently,
immunodeficient conditions. human papillomavirus, p. 631 occasional cases were identified among immunodeficient patients.
With the start of the AIDS epidemic, the incidence of PCP
MicroAssessment 28.2 increased rapidly, and it is still a common opportunistic infection
in AIDS patients who are not receiving preventive care.
Certain DNA viruses are strongly associated with development of
malignant tumors in patients with HIV disease. These viruses do Signs and Symptoms
not cause malignancy by themselves, but require the presence of
other conditions. The tumors all arise in a setting of increased cell
Many people are infected with the fungus that causes PCP, but
proliferation caused in part by the viruses. the infection is latent because it is kept under control by
the immune system. In immunocompromised individuals, how-
4. In what epidemiological forms does Kaposi’s sarcoma occur?
ever, the organism causes disease. The signs and symptoms of
5. What member of the herpesvirus family is associated with almost
Pneumocystis pneumonia typically begin slowly, with gradually
all of the B-cell lymphomas of the brain in AIDS patients?
increasing shortness of breath and rapid breathing. Fever is usu-
6. Would you expect mutations to arise in a population of rapidly
ally slight or absent, and only about half of the patients have a
dividing cells? +
cough, which is non-productive. As the disease progresses, a
dusky coloration of the skin and mucous membranes appears and
gradually worsens—this is caused by poor oxygenation of the
28.3 ■ Infectious Diseases blood, and can become fatal.
Learning Outcomes Causative Agent
8. Describe the significance of Pneumocystis jiroveci infection in Pneumocystis pneumonia is caused by Pneumocystis jiroveci,
people with HIV disease. a tiny yeastlike fungus (figure  28.10). The organism was for-
9. Outline the epidemiology of toxoplasmosis. merly classified as P. carinii and it is still widely known by that
10. Discuss the relationship between CMV disease and HIV disease. name (explaining the acronym PCP for this type of pneumonia).
11. Outline the pathogenesis of Mycobacterium avium complex P. jiroveci differs from many fungi in the chemical makeup of
infection in immunodeficient people. its cell wall, which makes it resistant to medications often used
against fungal pathogens. The organism forms cysts that have a
Immunodeficient people are susceptible to the same infections characteristic appearance, helping in its identification. It has not
as other people, but the disease is likely to be more severe and reliably been cultivated in vitro.
may even be fatal. Bacteria, viruses, fungi, protozoa, and even par-
asitic worms can cause life-threatening illness in those with AIDS. Pathogenesis
Preventing infections in the immunocompromised is therefore P.  jiroveci spores are easily inhaled into lung tissue. In experi-
very important. Vaccines should be given to immunocompetent mental infections, the spores attach to the alveolar walls, and the
 Part IV Infectious Diseases 709

TABLE 28.8 Pneumocystis Pneumonia

Signs and Gradual onset, shortness of breath, rapid
symptoms breathing, non-productive cough, slight or
absent fever, and dusky color of skin and
mucous membranes
Incubation 4 to 8 weeks
period
Causative agent Pneumocystis jiroveci, a tiny fungus
(previously known as P. carinii)
Pathogenesis Pneumocystis can result from reactivation of
latent infection or be newly acquired. Spores
of P. jiroveci enter the body when inhaled,
attach to alveolar walls, and multiply.
Alveoli fill with fluid, macrophages, and
P. jiroveci. The walls thicken, impairing O2
exchange.
Epidemiology P. jiroveci widespread in domestic and wild
animals as a latent lung infection, but the
30 μm source of animal and human infections is
unknown. Most humans become infected in
FIGURE 28.10 Fluorescent Antibody Stain of Pneumocystis early childhood. Disease arises in individuals
jiroveci The yellow circles are P. jiroveci cysts. with immunodeficiency; epidemics can
? Why is Pneumocystis pneumonia known by the acronym PCP? occur in hospitalized premature infants and
elderly nursing home residents.
Treatment Formerly leading cause of death in
and prevention those with AIDS, now usually prevented
alveoli fill with fluid, mononuclear cells, and masses of P. jiroveci by medication (e.g., trimethoprim-
cells in various stages of development. Later, the alveolar walls sulfamethoxazole) as soon as the CD4+
become thickened and scarred, preventing the free passage of O2. lymphocyte count drops to 200 cells/μl.
Same medication is used for treatment;
alternatives available. Medication is
Epidemiology continued for life, or until the CD4+ cell
Pneumocystis jiroveci is distributed worldwide. Most children are count rises and remains above 200 cells/μl as
infected with P. jiroveci at an early age. The infection is asymp- a result of HAART or other treatment of the
underlying immunodeficiency.
tomatic and is generally eliminated within a year. The source and
transmission of human infections are unknown. Most cases of
PCP occur in people with immunodeficiency, but it is uncertain
whether their disease is caused by reactivation of a latent infec-
tion, or new infection from inhalation of spores. Epidemics among Toxoplasmosis
hospitalized malnourished infants and elderly nursing home resi- Toxoplasmosis is a protozoan disease that rarely develops among
dents suggest airborne spread, and P. jiroveci has been detected in healthy people but can be a serious problem for those with malig-
indoor and outdoor air by using polymerase chain reaction (PCR). nant tumors, organ transplant recipients, fetuses, and people with
PCR, p. 227 HIV disease.

Treatment and Prevention Signs and Symptoms

PCP is most often treated with trimethoprim-sulfamethoxazole. Toxoplasmosis causes different signs and symptoms in immuno-
Alternative medications are given to people who cannot tolerate logically healthy people, fetuses, and those with immunodeficiency.
this medication because of its side effects—mainly rash, nausea, Most infections of immunocompetent people are asymptomatic,
and fever. For unknown reasons, people with HIV disease are but 10% to 20% develop signs and symptoms similar to those of
more likely to develop these side effects than others. Oxygen ther- infectious mononucleosis. They usually consist of sore throat, fever,
apy and steroids (to reduce inflammation that worsens symptoms) enlarged lymph nodes and spleen, and sometimes a rash. These signs
are also given and can significantly decrease mortality. After treat- and symptoms subside over weeks or months and do not require
ment for PCP, individuals with HIV disease must receive preven- treatment. Occasionally, life-threatening illness develops, affecting
tive medication indefinitely, or until they have a sustained rise in the heart or central nervous system. infectious mononucleosis, p. 680
CD4+ T-cell count to above 200 cells per microliter. Fetal toxoplasmosis can be acquired transplacentally if a
To prevent PCP, HIV-infected people are typically started on woman contracts the disease for the first time during her pregnancy.
medication as soon as they become immunodeficient, as indicated Development of the disease during the first trimester of pregnancy
by a CD4+ T-cell count below 200 per microliter, or development is the least common but most severe, often resulting in miscar-
of characteristic opportunistic diseases. The main features of riage or stillbirth. Babies born live may have serious birth defects
Pneumocystis pneumonia are presented in table 28.8. including small or enlarged heads, and lung and liver damage.
710 Chapter 28 HIV Disease and Complications of Immunodeficiency

Later, these babies can develop seizures or exhibit

intellectual disability. However, almost two-thirds of
fetal toxoplasmosis cases occur during the last trimester
of pregnancy, and the effects on the fetus are usually
less severe. Most of these infants appear normal at
birth, although later in life, retinitis—infection of the
retina, the light-sensitive part of the eye—may occur.
This manifests itself as recurrent episodes of pain, sen-
sitivity to light, and blurred vision, usually involving
(a) 15 μm
only one eye. Intellectual disability and epilepsy may
develop. Congenital toxoplasmosis does not occur in fetuses of
Animals with
women who have had previous exposure to T. gondii, most likely cysts in tissue
because of immune protection.
Toxoplasmosis in immunodeficient people is often life-
threatening. It commonly occurs as a reactivation of a latent
T. gondii infection, and it is associated with various signs and
symptoms including those similar to infectious mononucleosis.
In more than half the cases, the infected person gets encephalitis
(inflammation of the brain meninges), manifested by confusion, Immature oocyst
weakness, impaired coordination, seizures, stiff neck, paralysis,
and coma. They can also develop brain masses much like tumors, Sporocysts
Contaminated
which cause headaches and other neurological signs and symp- food ingested
toms such as unstable walking gait, seizures, sensory loss and by animals Sporozoite
weakness. Infection of the retina is also common.
Causative Agent Mature oocyst

Toxoplasmosis is caused by Toxoplasma gondii, a tiny, banana- Changing

litter box
shaped protozoan that infects many animals, including household
pets, pigs, sheep, cows, rodents, and birds (figure  28.11a). The
life cycle of T. gondii is shown in figure  28.11b. The definitive
host, in which the organism reproduces sexually, is the domestic
cat or other felines (such as bobcats). The protozoan reproduces
in the cat’s intestinal lining, resulting in many progeny. Some of
these spread throughout the body, while others differentiate into
male and female gametes (sexual forms). Gametes fuse, forming
thick-walled oocysts that are shed in the cat’s feces. Millions of Infected raw
or undercooked Humans ingest
the oocysts are released daily, generally over a period of 1 to oocytes in
meat
3 weeks. In soil, the oocysts undergo further development over contaminated
1 to 5 days into an infectious form containing two sporocysts, each food, or from
unclean hands
with four sporozoites. These can remain viable for up to a year,
contaminating soil and water and, secondarily, hands and food. (b)
The oocysts are infectious for cats and other animals, including
humans. In general, the cats recover from the acute infection and
do not shed oocysts again.
When non-feline animals ingest the oocysts, the sporozoites
emerge from the oocysts and invade the cells of the small intestine. Cyst
They do not undergo a sexual cycle. The intestinal infection spreads
by the lymphatics and blood vessels throughout the tissues of the
host, infecting cells of the heart, brain, and muscles. As host immu-
T. gondii
nity develops, multiplication slows, and a tough, fibrous capsule
forms, surrounding large numbers of a smaller form of T. gondii. Congenital infection
These capsules are called cysts (figure  28.11c), and they remain (c) 10 μm
viable for months or years. The life cycle is completed when a cat FIGURE 28.11 Toxoplasma gondii (a) Invasive forms. (b) Life
becomes infected by eating an animal with cysts in its tissues. cycle. Oocysts from cat feces and cysts from raw or inadequately
cooked meat can infect humans and many other animals. (c) Cyst in
Pathogenesis tissue.
The organisms enter the body by ingestion of oocysts or under- ? Why does eating undercooked meat increase the risk of
cooked meat containing tissue cysts. T. gondii infects any kind contracting toxoplasmosis?
 Part IV Infectious Diseases 711

of cell except red blood cells. The organism produces an enzyme

TABLE 28.9 Toxoplasmosis
that alters the host cell membrane and aids entry into the cell. The
infected cells are destroyed by the proliferating T. gondii. This pro- Signs and In healthy individuals: sore throat, fever,
cess is normally brought under control by the immune response of symptoms enlarged lymph nodes, rash; with fetal
infections: miscarriage, stillbirth, birth
the host—tissue cysts develop, and infected humans usually show
defects, epilepsy, intellectual disability,
few, if any, signs and symptoms. In patients with immunodefi- retinitis; in immunodeficient individuals:
ciency, however, infection can be widespread and uncontrolled, confusion, poor coordination, weakness,
producing many areas of tissue necrosis. The disease process can paralysis, seizures, and coma
result from a newly acquired infection, or from reactivation of Incubation Usually indeterminate
latent infection that occurs with declining immunity. period
Causative agent Toxoplasma gondii, a protozoan infectious
Epidemiology for most warm-blooded animals. Sexual
Toxoplasma gondii is distributed worldwide, but it is less com- reproduction occurs in the intestinal
epithelium of cats, the definitive hosts.
mon in cold or hot, dry climates. Human infection is widespread.
Infected cats discharge oocysts with their
Serological surveys show the infection rate increases with age. feces. Ingested organisms are released
Most infections are acquired when a person eats or drinks some- from the oocysts, multiply rapidly, spread
thing contaminated with oocysts, or inhales contaminated dust. throughout the body. As immunity develops,
Gardening in areas where young, stray cats that eat rodents and infected cells become filled with the
organisms, resulting in tissue cysts, which
birds live can result in T. gondii contamination of hands or vege-
remain viable and infectious for the lifetime
tables. Eating rare meat is risky because it can contain tissue cysts. of the animal.
Pathogenesis Organisms penetrate host cells causing
Treatment and Prevention necrosis. With development of immunity
Toxoplasmosis is treated with drugs such as pyrimethamine with cell destruction stops, tissue cysts develop.
sulfadiazine, which are related to trimethoprim-sulfamethoxazole Most healthy individuals have few or no
(used to treat PCP). Alternatives are available. All HIV-infected symptoms unless the number of ingested
organisms is very large. Organisms released
patients and those about to receive immunosuppressant medica- from tissue cysts if immunity becomes
tions are tested for antibody to T. gondii. A positive test indicates impaired.
they have latent infection, and those with positive tests are given
Epidemiology Occurs worldwide, less common in cold
prophylactic trimethoprim-sulfamethoxazole if they have fewer or dry locations. Infection acquired by
than 100 CD4+ T cells per microliter. PCP, p. 708 ingesting oocysts from cat feces, or eating
T. gondii infection can be avoided by washing hands after inadequately cooked meat.
touching raw meat, soil, or cat litter. Meat, especially lamb, pork, Treatment Treatment: pyrimethamine with
and venison (deer meat), should be cooked thoroughly. Fruits and and prevention sulfadiazine, or alternative medication.
vegetables should be washed before eating. Litter boxes should be Prevention: avoiding foods potentially
cleaned regularly, before oocysts have a chance to mature. Cats contaminated with oocysts from cat feces,
and not eating inadequately cooked meat.
should not be allowed to hunt birds and rodents, nor should they Trimethoprim-sulfamethoxazole is given
be fed undercooked or raw meat. These measures are especially to immunodeficient persons with CD4+ T
important for pregnant women and immunodeficient people. The lymphocyte counts below 100 cells/μl if
main features of toxoplasmosis are presented in table 28.9. they have antibodies to T. gondii indicating
latent infection.

Cytomegalovirus Disease
Cytomegalovirus (CMV) is a member of the herpesvirus family. characterized by jaundice, enlarged liver, anemia, eye inflamma-
Like other herpesviruses, CMV is commonly acquired early in life tion, and birth defects. The vast majority of infected infants appear
and then remains latent. With immunodeficiency, the infection healthy at birth, but up to a quarter of them develop hearing loss,
reactivates and can cause severe illness. mental handicap, or other abnormalities later in life.
In immunodeficient people, latent CMV infection can reac-
Signs and Symptoms tivate, causing a variety of signs and symptoms. Retinitis, lead-
Signs and symptoms of cytomegalovirus disease follow a ing to blindness, is one of the most common complications of
pattern similar to that of toxoplasmosis. Acute infections in reactivated CMV disease. Other signs and symptoms include
immunologically healthy people are usually asymptomatic, but fever, loss of appetite, painful joints and muscles, rapid and diffi-
adolescents and young adults sometimes develop illness that cult breathing, ulcerations of the gastrointestinal tract with bleed-
resembles infectious mononucleosis—fever, fatigue, and enlarged ing, lethargy, paralysis, dementia, coma, and death.
lymph nodes and spleen that can persist for weeks or months.
If a woman develops an acute CMV infection during Causative Agent
pregnancy, her fetus can be severely damaged. This condition Human cytomegalovirus is an enveloped, double-stranded DNA
is known as congenital cytomegalic inclusion disease and is virus that looks like other herpesviruses on electron micrographs
712 Chapter 28 HIV Disease and Complications of Immunodeficiency

but has a larger genome. The virus name (cyto for “cell” and
megalo for “large”) derives from the fact that cells infected
with it are two or more times the size of uninfected cells.
Infected cells show a large, intranuclear inclusion body
surrounded by a clear halo, inspiring its description as
an “owl’s eye” (figure 28.12). There are many different
strains of CMV, and like other herpesviruses, CMV can
lyse infected cells or can become latent with possible
later reactivation. Hemorrhage

Pathogenesis
Cytomegalovirus disease affects a wide variety of tissues,
including the eye, central nervous system, lung, and liver
(figure 28.13). Once the virus enters the host cell, the viral genome
can exist in a latent, non-infectious form or a slowly replicating form, (a)
or it can direct a productive infection. productive infection, p. 310
Control of viral gene expression depends partly on the type
of cell infected. In monocytes, low numbers of infectious viral
particles are produced. In T and B lymphocytes, some CMV genes
are expressed, but the viral genome is not replicated nor are new
viral particles produced. Integration of viral DNA into the host
cell genome probably does not occur. If CMV-infected T cells
are also infected with HIV, however, productive CMV infection
occurs. This involves a wide variety of tissues, causing necrosis. (b)
CMV can be transmitted by transplanted organs and blood FIGURE 28.13 Cytomegalovirus (CMV) Retinitis Photograph
transfusions. This indicates that virus-producing cells are present of a normal retina (a) compared with a CMV-infected retina (b).
or that non-producing cells in the blood or organ start producing ? What does CMV retinitis commonly lead to in people with AIDS?
infectious CMV under conditions of immune suppression. Cell-
mediated immunity probably plays a role in suppressing produc-
tion of infectious virus as well as in lysis of infected cells. In cells
infected with CMV, however, transfer of MHC molecules to the AIDS, organ transplants, and other immunodeficient states. Also,
cell surface is impaired and, therefore, CMV antigens are not rec- immunodeficient people are highly susceptible to newly acquired
ognized as being “foreign.” CMV infection is associated with an infection. MHC molecules, p. 369
increase in CD8+ cells and a decrease in CD4+ cells, thus enhanc-
ing the effect of HIV infection. Latent infections activate with Epidemiology
CMV is found worldwide. One U.S. study found that more than
Infected cells 50% of adults, ages 18 to 25 years, and more than 80% of people
over age 35 had been infected. Infection is lifelong. Infants born
with CMV infection and those who acquire it shortly after birth
excrete the virus in their saliva and urine for months or years. Virus
is found in saliva, semen, and cervical secretions. Unprotected sex
is a common mode of transmission in young adults. Up to 15% of
pregnant women secrete the virus, and 1% of newborn infants have
CMV in their urine. Breast milk, blood, and tissue transplants may
contain CMV and can be responsible for transmission.

Treatment and Prevention

The severity of CMV disease can be reduced by taking the antivi-
ral medications ganciclovir and foscarnet in combination. These
medications inhibit DNA polymerase, thus affecting replication
of the virus. Both medications have serious side effects, however,
mainly bone marrow suppression and kidney damage.
20 µm There is no approved vaccine for preventing cytomegalovirus
FIGURE 28.12 Cytomegalovirus Infection Infected cells are two disease. The risk of sexual transmission is effectively reduced by
to four times normal size. using condoms. Tissue and blood donors can be screened for anti-
? Why are CMV-infected cells described as having an “owl’s-eye” body to CMV; those who have anti-CMV antibody are assumed to
appearance? be infected and should not donate to those lacking CMV antibody.
 Part IV Infectious Diseases 713

The incidence of CMV retinitis in people with HIV disease can Signs and Symptoms
be reduced by half if ganciclovir is given. A ganciclovir implant The vast majority of MAC infections in immunologically healthy
designed to release the medication into the eye over a long period people are asymptomatic. However, elderly people, especially those
also delays the progression to blindness. The main features of with underlying lung disease from smoking and those with alcohol-
cytomegalovirus disease are presented in table 28.10. ism, develop a chronic, productive cough. They may also develop
lung lesions resembling those of tuberculosis. Children sometimes
Mycobacterial Diseases develop chronic lymph node enlargement on one side of their neck,
easily treated by surgical removal of the affected nodes. Patients with
In most immunocompetent people, initial exposure to AIDS and other severe immunodeficiencies have slowly progressing
Mycobacterium tuberculosis (the cause of tuberculosis) results in signs and symptoms ranging from chronic productive cough to fever,
an asymptomatic infection that is controlled by the immune sys- drenching sweats, marked weight loss, abdominal pain, and diarrhea.
tem and becomes latent. However, defects in cell-mediated immu-
nity caused by AIDS and other immunodeficiencies often result in Causative Agents
reactivation of latent tuberculosis, leading to active tuberculosis
MAC consists of groups of more than 24 strains of two closely
disease, which may be fatal. tuberculosis, p. 502
related acid-fast species—M. avium and M. intracellulare. The
M. tuberculosis is not the only mycobacterium that causes
strains are distinguishable by serological tests, optimum growth
disease in immunodeficient people, although it is the most com-
temperature, and host range. Their growth rate is almost as slow
mon. This section discusses disease caused by organisms of
as that of M. tuberculosis, but they are easily distinguished from
Mycobacterium avium complex (MAC), mycobacterial opportun-
this organism by using biochemical tests and nucleic acid probes.
ists that complicate immunodeficiency diseases.
Pathogenesis
MAC organisms enter the body via the lungs and the gastrointestinal
TABLE 28.10 Cytomegalovirus Disease
tract. They are phagocytized by macrophages, but resist destruction
Signs and Signs and symptoms rare in because they inhibit acid production in the phagosome. Surviving
symptoms immunocompetent individuals, but may organisms multiply within phagocytes and are carried by the blood-
resemble infectious mononucleosis if they
stream to all parts of the body. When cell-mediated immunity is
develop. Infection of the mother during
pregnancy can result in disease of the intact, most organisms are destroyed, and disease is localized. With
newborn. Immunocompromised individuals immunodeficiency, the disease spreads throughout the body. In these
may experience blindness, lethargy, patients, there is persistent bacteremia, occasionally with counts as
dementia, coma, and brain damage. high as 1 million bacteria per milliliter of blood. The small intestine
Incubation In immunocompetent adults, 20 to 60 days contains macrophages packed with high numbers of the bacteria
period (figure 28.14). Despite the presence of enormous numbers of the bac-
Causative agent Cytomegalovirus, a member of the teria, there is little or no inflammatory reaction, and the clinical effect
herpesvirus family. is a slow decline in the patient’s well-being rather than a quickly
Pathogenesis Many tissues susceptible to infection, lethal effect. One cause of deterioration may be activation of HIV
damage, especially eyes, brain, and liver. replication when HIV-infected macrophages are infected with MAC.
CMV latent infection can reactivate, produce
infectious virions, tissue necrosis. CD4+ T
MAC bacteria
lymphocyte count is depressed and thus can
enhance HIV disease.
Epidemiology Common worldwide; lifelong infection. More
than 50% of 18- to 25-year-olds are infected.
Infants with congenital infections and those
infected shortly after birth shed the virus
for months or years. Body fluids, including
breast milk, blood, urine, semen, and vaginal
secretions, can transmit the disease.
Treatment Treatment: ganciclovir plus foscarnet.
and prevention Prevention: No vaccine available. Condoms
decrease transmission. CMV-negative
immunodeficient people advised to wash
their hands following contact with bodily
fluids or feces. Blood and tissue transplants
are tested for CMV before being given to
CMV-negative individuals. The antiviral
medication ganciclovir is considered for
immunodeficient individuals who have 10 μm
antibody to CMV and whose CD4+ T FIGURE 28.14 MAC Infection Acid-fast stain showing
lymphocyte count falls below 50 cells/μl. massive numbers of MAC bacteria infecting cells.
? What species are involved in MAC infection?
714 Chapter 28 HIV Disease and Complications of Immunodeficiency

TABLE 28.11 MAC Disease Epidemiology

MAC organisms are widespread and have been found in food,
Signs and Usually asymptomatic in healthy people. water, soil, and dust. Some strains are important pathogens of
symptoms Children can develop chronic localized
chickens and pigs. In AIDS patients, they are the most common
enlargement of lymph nodes. Can cause
chronic cough similar to tuberculosis in the bacterial cause of generalized infection. It is not known whether
elderly. Cough, fever, sweating, marked infection in AIDS patients is mostly newly acquired or a reacti-
weight loss, abdominal pain, and diarrhea in vation of latent disease. Most infections are from environmental
those with severe immunodeficiencies. sources rather than from person-to-person spread.
Incubation period Usually indeterminate
Causative agents A group of mycobacterial strains in the
species M. avium and M. intracellulare Treatment and Prevention
Pathogenesis MAC bacteria enter the body via lungs If MAC bacteremia develops, patients must be given two or
or gastrointestinal tract, are taken up more medications together, such as clarithromycin with etham-
by macrophages. In immunocompetent
individuals, most are destroyed and infection
butol. There are no effective measures to prevent exposure to
is controlled by cell-mediated immunity. In MAC organisms. For AIDS patients whose CD4+ count is below
severe immunodeficiency the bacteria resist 50 cells per microliter, the antibacterial medication clarithromycin
phagocytosis, are carried by macrophages is recommended to help prevent MAC disease. The main features
throughout the body, and grow to enormous of MAC disease are presented in table 28.11.
numbers in tissues but cause little or no
inflammatory reaction.
The key features of the diseases covered in this chapter are high-
Epidemiology MAC organisms are widespread in food,
lighted in the Diseases in Review 28.1 table.
water, soil, and dust. MAC disease of
immunodeficient persons could result from
environmental sources, or activation of latent MicroAssessment 28.3
infection.
Infectious diseases are a major threat to individuals with
Treatment No proven measures to prevent exposure to
immunodeficiency. They get the same infectious diseases that
and prevention MAC bacteria. The antibiotic clarithromycin
afflict healthy people, but these can be life-threatening. In addition,
is used to prevent MAC disease in severe
immunocompromised people are much more susceptible to
immunodeficiency. If MAC bacteremia
develops, two or more medications, such as
reactivation of latent infections and organisms of low virulence that
clarithromycin plus ethambutol, are effective. are generally harmless to others. Prevention of infectious diseases in
AIDS patients is important for improving the quality and duration of
their lives.
MicroByte 7. What is the most common opportunistic infection of
MAC-infected people may contain 100 billion of the bacteria per immunodeficient people?
gram of tissue. 8. Which organisms are involved in MAC bacteremia? +

FUTURE CHALLENGES 28.1

AIDS and Poverty
In the United States and other economically AIDS sufferers because they cannot afford them. The humanitarian, social, and economic
advanced countries, antiviral medications have Only about 5.2 million of the 15 million people implications of uncontrolled HIV disease are
contributed significantly to reducing mother- needing anti-HIV medications are receiving enormous. Unimaginable suffering; large
child transmission of HIV, to delaying pro- them. Although HIV disease is spreading rapid- numbers of starving, uneducated street chil-
gression of HIV disease to AIDS, and to ly in India and China—two countries that dren; loss of productive work years; spread of
improving the quality and duration of life together contain more than one-third of the other infectious diseases; breakdown of public
of AIDS sufferers. Newer drugs have also world’s population—and in the Caribbean, health; and setbacks for economic develop-
decreased transmission of HIV somewhat, Southeast Asia, and Eastern Europe, the situa- ment are possible consequences that can affect
thereby slowing the progress of the epidemic. tion is the most desperate in African countries neighboring countries and the rest of the world.
The cost in the United States, while high, south of the Sahara Desert. Only 10% of the The challenge is urgent: to find effec-
amounts to less than 1% of the country’s total world’s population lives in this area, but two- tive HIV control measures, especially for the
healthcare expenditure. thirds of the people are infected with HIV. About world’s poor.
Antiviral medications, however, have 75% of the region’s young people (ages 15 to
provided little benefit to most of the world’s 24) infected with HIV are girls or young women.
Diseases in Review 28.1
HIV Disease and Complications Caused by Malignant Tumors
and Infectious Diseases

Disease Causative Agent Comment Summary Table

HUMAN IMMUNODEFICIENCY VIRUS AND AIDS

HIV disease Human Acute retroviral syndrome marks the early stage; period Table 28.6, p. 705
immunodeficiency virus of clinical latency can last for many years; AIDS begins
(HIV) as the person becomes increasingly immunodeficient;
HAART lowers viral set point, extending clinical latency;
most antiretroviral drugs target reverse transcriptase or
protease; vaccine research has been hampered by the lack
of good animal model, HIV’s high mutation rate, and HIV’s
ability to avoid humoral and cellular immune responses.

MALIGNANT TUMORS THAT COMPLICATE ACQUIRED IMMUNODEFICIENCIES

Kaposi’s sarcoma Human herpesvirus-8 These tumors arising from blood or lymphatic vessels
(HHV-8, also called are characterized by red, brown, purple, or black
KSHV) papular nodules on the skin or other body sites; in
immunocompetent people, the virus typically causes an
asymptomatic infection that becomes latent.
B lymphocyte tumors Epstein-Barr virus (EBV) Latent EBV reactivates as a result of HIV infection; the role
of EBV in tumor formation is not well understood.
Cervical and anal Certain strains of HPV replication increases with decreasing cell-mediated
carcinoma human immunity; HPV blocks normal cellular gene that controls
papillomaviruses (HPVs) cell growth.

INFECTIOUS DISEASES THAT COMPLICATE ACQUIRED IMMUNODEFICIENCIES

PROTOZOAN DISEASES

Pneumocystis Pneumocystis jiroveci The most common opportunistic infection in AIDS patients; Table 28.8, p. 709
pneumonia (PCP) in immunocompetent people, the fungus typically causes
an asymptomatic infection that becomes latent.
Toxoplasmosis Toxoplasma gondii Latent T. gondii reactivates as a result of AIDS, often Table 28.9, p. 711
resulting in encephalitis; acute infections in pregnant
women can damage the fetus, resulting in miscarriages
or severe birth defects.

VIRAL DISEASE

Cytomegalovirus Cytomegalovirus (CMV) Latent CMV reactivates as a result of AIDS, often resulting Table 28.10, p. 713
disease in retinitis, which causes blindness; infection causes a
variety of other signs and symptoms; acute infections in
pregnant women can damage the fetus.

BACTERIAL DISEASES

Mycobacterial Mycobacterium species Tuberculosis is a significant problem in AIDS patients; Table 28.11, p. 714
diseases a group of opportunists called Mycobacterium avium
complex (MAC) can cause similar signs and symptoms in
AIDS patients.
716 Chapter 28 HIV Disease and Complications of Immunodeficiency

Summary
AIDS in the brain. Strong evidence suggests that Epstein-Barr virus plays a
causative role in these tumors.
Since AIDS was first recognized in 1981, it has claimed millions of
lives in the United States, and is a leading cause of death in people 25 Cervical and Anal Carcinoma
to 44 years of age. Worldwide, over 34 million people are infected with There is an increased rate of anal, genital, and cervical carcinoma in
an AIDS-causing virus (figure  28.1). In sub-Saharan Africa, AIDS is people with HIV disease. These cancers are strongly associated with
now the number one cause of death, surpassing malaria; it is spreading human papillomaviruses (HPVs), transmitted by sexual intercourse.
rapidly in India and into China.

28.1 ■ Human Immunodeficiency Virus (HIV) 28.3 ■ Infectious Diseases

Infection and AIDS Infections that occur in healthy individuals also occur and produce more
“AIDS-defining conditions,” including serious infections by agents that severe disease in those with immunodeficiency.
normally have little virulence, usually reflect immunodeficiency and
were especially useful “markers” in early studies of the AIDS epidemic Pneumocystis pneumonia (PCP) (table 28.8, figure 28.10)
(table 28.1). Before effective preventive regimens were developed, PCP was one
of the most common causes of death among AIDS patients. Signs and
HIV Disease (tables 28.2–28.7, figures 28.2–28.7)
symptoms develop slowly, with gradually increasing shortness of breath
Acquired immunodeficiency syndrome (AIDS) is a late manifesta- and rapid breathing; patients can die from lack of O2. The causative
tion of human immunodeficiency virus (HIV) disease. “Flulike” agent, Pneumocystis jiroveci, a tiny fungus, is widespread in humans,
or infectious mononucleosis-like signs and symptoms representing generally causing asymptomatic infections that become latent in immu-
the acute retroviral syndrome (ARS) often occur 6 days to 6 weeks nocompetent people.
after infection by HIV; these subside without treatment. HIV disease
progresses asymptomatically for years, although HIV can still be Toxoplasmosis (table 28.9)
transmitted to others during this time. The asymptomatic period ends Toxoplasmosis is caused by Toxoplasma gondii, a tiny, protozoan pres-
with the onset of immunodeficiency (AIDS), marked by opportunis- ent worldwide. This organism reproduces in the intestinal epithelium of
tic or reactivated infections, and certain tumors caused by infectious cats but can infect humans and other vertebrates. People contract toxo-
agents. HIV disease and AIDS are caused by human immunodeficiency plasmosis by ingesting oocytes, which are discharged in the feces of
virus, a single-stranded RNA virus belonging to retrovirus family acutely infected cats, become infectious in soil, and contaminate food,
(table 28.2, figure  28.3). HIV disease is spread mainly by sexual inter- water, and fingers (figure  28.11). Toxoplasmosis is rare among healthy
course, by blood or blood products, and from mother to fetus or newborn people, and if it does occur is usually asymptomatic. Signs and symp-
(tables 28.4, 28.5). Different kinds of cells can be infected by HIV, nota- toms that may develop mimic those of infectious mononucleosis and
bly two kinds vital to the immune system—helper T lymphocytes and usually resolve without treatment. This disease, however, can be a seri-
macrophages (figure 28.4). Inside the host cell, a DNA copy of the viral ous problem for people with AIDS, who frequently develop encephalitis
genome is made by reverse transcriptase, a complementary DNA strand and brain masses that may be fatal.
is made, and the resultant double-stranded DNA is inserted into the host
genome as a provirus by viral integrase (figures 28.5, 28.6). Antimicrobial Cytomegalovirus Disease (table 28.10)
medications are used to prevent and treat opportunistic infections, and Cytomegalovirus (CMV) is a common cause of impaired vision in
highly active antiretroviral therapy (HAART) is used to maintain a people with AIDS (figure 28.13). It can also cause fever, gastrointestinal
low HIV set point and manage HIV disease (tables 28.3, 28.7; figure 28.7). bleeding, mental dullness, and blindness in these individuals. CMV is
an enveloped, double-stranded DNA virus; infected cells are enlarged
HIV Vaccine Prospects and have an “owl’s-eye” appearance (figure 28.12). The virus occurs in
There are no approved vaccines for HIV at this time. breast milk, semen, and cervical secretions, and in saliva and urine of
infected infants. No vaccine is available. Condoms decrease the risk of
sexual transmission. The antiviral medication ganciclovir can be given
COMPLICATIONS OF ACQUIRED to prevent CMV retinitis.
IMMUNODEFICIENCIES
Mycobacterial Diseases (table 28.11)
28.2 ■ Malignant Tumors Latent Mycobacterium tuberculosis may reactivate and cause active
Kaposi’s Sarcoma (figure 28.9) tuberculosis disease in immunocompromised people. M. avium
Kaposi’s sarcoma is a tumor that arises from blood or lymphatic ves- complex (MAC) organisms also commonly cause lung infections.
sels. The incidence of the tumor is markedly increased among immu- Immunodeficient patients may have fever, drenching sweats, severe
nodeficient individuals. Infection by human herpesvirus-8 (KSHV) weight loss, diarrhea, and abdominal pain. MAC organisms resist
appears to be involved in tumor development. phagocytic destruction and are carried to all parts of the body in macro-
phages, multiplying without restriction, and producing massive numbers
B Lymphocyte Tumors of the organisms in blood, intestinal epithelium, and other tissues (fig-
Lymphomas are malignant tumors that arise from lymphocytes. Both ure 28.14). No effective preventive measures are available. Prophylactic
B and T lymphocytes can give rise to lymphomas in immunodeficient medication is advised for severely immunodeficient patients, but it can
people, but B-cell lymphomas are more common, and these often arise fail to prevent infection.
 Part IV Infectious Diseases 717

Review Questions
Short Answer 7. Which of the following types of cells can be infected by HIV?
1. What is the main symptom of patients with lymphadenopathy syn- a) Helper T cells
drome (LAS)? b) Cytotoxic T cells
2. Which cells of the immune system are prime targets of HIV? c) B lymphocytes
3. What role do asymptomatic people with HIV disease play in the d) CD 8+ cells
epidemiology of AIDS? e) All of the above
4. Why might the infant son of a hemophiliac man develop AIDS 8. All of the following are HIV antigens except
when the son’s parents were strictly monogamous non-abusers of a) CD4. b) TM. c) RT. d) MA. e) CA.
drugs? 9. Which of the following is a cause of helper T-cell death in HIV
5. Give two reasons it is a good idea to know whether you are disease?
infected with HIV. a) Replication of HIV lyses the cell.
6. What are the three main types of malignant tumors that complicate b) Infected cells are destroyed by cytotoxic T cells (TC).
HIV disease? c) Infected cells are attacked by natural killer cells.
7. How do physicians prevent pneumocystis in AIDS patients? d) Cells are killed by fusion and syncytium formation.
8. In AIDS patients with toxoplasmosis, which part of the body is e) All of the above
affected in more than half the cases? 10. Highly active antiretroviral therapy (HAART) is less than ideal
9. Name a feared complication of cytomegalovirus infection in AIDS because
patients. a) it does not eliminate latent HIV infection.
10. Where in an AIDS patient’s surroundings might MAC organisms b) its cost is too great for the majority of AIDS sufferers.
be found? c) it often has severe side effects.
d) some HIV strains are resistant to it.
Multiple Choice
e) All of the above
1. HIV can be spread by all of the following except
a) blood products. b) hypodermic syringes. c) insect bites. Applications
d) sexual intercourse. e) organ transplants. 1. An epidemiologist from the CDC was presenting a report on the
2. All of the following signs and symptoms are characteristic of the status of AIDS to a congressional committee. In concluding her
AIDS-related complex (ARC) except remarks, she noted that from an epidemiological perspective it was
a) fever. b) fatigue. c) diarrhea. more important to focus on HIV infection than on AIDS, and urged
d) blindness. e) weight loss. that the Congress consider redirecting funding of AIDS research to
reflect this fact. What was the rationale for her request?
3. Which one of the following is true of Kaposi’s sarcoma?
2. A historian researching the influence of society on the spread of
a) KSHV is necessary for development of the tumor.
communicable disease began to speculate on what it would be like
b) HIV-1 is necessary for development of the tumor. if AIDS had appeared at a different time. What differences might
c) Both KSHV and HIV-1 are necessary for development of the one expect, for example, if AIDS had appeared in 1928 instead of
tumor. 1978?
d) KSHV alone is sufficient for development of the tumor.
e) Both KSHV and HIV-1 together are sufficient for the tumor to Critical Thinking +
develop. 1. Vaccines have effectively prevented many viral diseases. Attempts
4. All of the following are HIV accessory genes except over many years to develop an effective vaccine against HIV dis-
a) tat. b) env. c) vpr. d) rev. e) vpu. ease and AIDS, however, have so far met with little success. Why
5. When was AIDS first recognized as representing a new disease? is this so?
a) 1973 b) 1959 c) 1981 d) 1989 e) 1999 2. Why is reverse transcriptase needed in order for HIV to become a
provirus?
6. All of the following are AIDS-defining conditions except
a) influenza.
b) herpes simplex of the esophagus.
c) Pneumocystis jiroveci pneumonia.
d) invasive cancer of the uterine cervix.
e) Kaposi’s sarcoma.
29 Microbial Ecology
K
KEEY TE
Y T ER
RMM SS
Consumers Organisms that
eat primary producers or other
consumers.
Decomposers Organisms that
community characterized by
distinct layers of different groups
of microbes that together make up
a thick, dense, highly organized
structure.
digest the remains of primary
producers and consumers. Nitrogen Fixation Conversion of
nitrogen gas to ammonia.
Eutrophic An environment that
is nutrient rich, supporting the Oligotrophic A nutrient-poor
excessive growth of algae and other environment.
organisms. Primary Producers Organisms
Hydrothermal Vents Undersea that convert CO2 into organic
geysers that spew out mineral-laden compounds, sustaining other
hot water. life-forms.
Hypoxic An environment very low Rhizosphere Zone around plant
in dissolved O2. roots containing organic materials
exuded by the roots.
Microbial Mat A type of microbial

Farming relies on the activities of microorganisms.

M
 
A Glimpse of History icrobes cycle nutrients, maintain fertile soil, and decom-
pose wastes and other pollutants. Without microbial
For centuries, farmers have understood that growing the same crop on
activities, life on earth could not survive. People would
the same piece of land year after year reduces the crop yield. Allowing a
quickly become buried by the tons of wastes we generate, and
field to lie unplanted for one or more seasons lets wild plants grow, and
these appear to rejuvenate the soil. It was not until the late nineteenth cen- nutrients would be depleted, halting growth and reproduction. In
tury that scientists began to discover why this was so. They isolated soil view of the crucial functions microorganisms perform, it seems
microorganisms associated with certain plants that could take nitrogen we should know a great deal about the diverse microbial species
from the air and transform it into forms other organisms could use. This that inhabit our surroundings. Quite the opposite is true, however,
process is called nitrogen fixation, and the nitrogen is said to be “fixed.” as less than a mere 1% have been successfully grown in culture.
Although many scientists have worked for years to understand Even if all microorganisms could be cultivated in the labo-
how microorganisms fix nitrogen, one scientist from the Netherlands, ratory, the information gained might not accurately reflect their
Martinus Beijerinck, stands out as an early contributor in these studies. In role in the environment. In the laboratory, organisms are grown
the late 1880s, he isolated a bacterium from inside the nodules that form as pure cultures under controlled conditions that ensure optimal
on the roots of legumes (plants that bear seeds in pods). Russian micro-
growth. In nature, however, organisms generally grow as mem-
biologist Sergei Winogradsky then showed that the bacterium forms a
bers of mixed communities in poorly defined and often chang-
symbiotic relationship with the legumes. We now know that the bacterial
ing conditions. Nutrients are normally in short supply, limiting
cells in the nodules fix nitrogen, and they are in a group called rhizobia.
growth. Thus, with respect to environmental microbiology, results
rhizobia, p. 269
Beijerinck made major contributions to several areas of microbiol- obtained in the artificial setting of the laboratory, although useful,
ogy. He worked on yeasts, plant viruses such as tobacco mosaic virus, must be interpreted with caution. bacterial growth in nature, p. 84
and plant galls (tumors). He was described as a “keen observer” who Chapters 4 and 11 discussed prokaryotic growth in nature,
was able “to fuse results of remarkable observations with a profound and and microbial diversity. This chapter will expand on some of those
extensive knowledge of biology and the underlying sciences.” This abil- concepts, focusing on activities of microorganisms that make
ity was undoubtedly partly responsible for the great success of his work. them essential to life.

718
 Part V Applied Microbiology 719

29.1 ■ Principles of Microbial called the biosphere. Within the biosphere, ecosystems vary both
in biodiversity (number and variety of species present and their
Ecology evenness of distribution) and biomass (the weight of all organ-
isms present). Microorganisms play a major role in most ecosys-
Learning Outcomes tems, and many ecosystems host microbes unique to themselves.
1. Describe the roles of primary producers, consumers, and The role an organism plays in a particular ecosystem is called its
decomposers. ecological niche.
2. Describe how some microbes are able to grow in low-nutrient The environment immediately surrounding an individual
environments. microbe—the microenvironment—is most relevant to that cell,
3. Compare and contrast microbial competition and antagonism. but because microorganisms are so small, the microenvironment
4. Describe how environmental changes can result in alterations in a is difficult to identify and measure. The more readily measured
microbial community. gross environment—the macroenvironment—may be very dif-
5. Describe the structural organization of a microbial mat. ferent from the microenvironment. Consider a bacterial cell living
6. Describe three methods researchers use to better understand within a biofilm (see figure 4.3); growth of aerobic organisms in
complex microbial communities. the biofilm can deplete O2, creating microzones where obligate
anaerobes can grow. Fermenters can produce organic acids that
Ecology is the study of the relationships of organisms—plant may then be metabolized by other organisms in the film, and vari-
and animal—to each other and to their environment. Likewise, ous growth factors can be transferred between organisms as well.
microbial ecology is the study of the relationships of microor- Thus, certain microorganisms that might seem unexpected in a
ganisms to each other and to their environment. Living organisms given macroenvironment actually thrive there because of micro-
interact with one another in symbiotic relationships (commen- environments. biofilm, p. 84 growth factor, p. 93
salism, mutualism, and parasitism), described in chapter 16.
symbiosis, p. 381 Nutrient Acquisition
Organisms in a given area, the community, interact with each
Organisms are categorized according to their trophic level (source
other and the non-living environment, forming an ecological sys-
of food), which is intimately related to the cycling of nutrients.
tem, or ecosystem. Major ecosystems include the oceans, rivers
There are three general trophic levels (figure 29.1):
and lakes, deserts, marshes, grasslands, forests, and tundra. Each
ecosystem possesses a certain spectrum of organisms ■ Primary producers. These are autotrophs; they convert CO2
and characteristic physical conditions. The region into organic materials. Producers include both photoauto-
of the earth inhabited by living organisms is trophs (use sunlight for energy) and chemolithoautotrophs

Sun FIGURE 29.1 Trophic Levels in an Ecosystem

H2S, NH3 ? Which organisms in the diagram are autotrophs?
and other
reduced
inorganic
chemicals
Tertiary consumer

Radiant Chemical Secondary consumer

energy energy

Primary consumer

Primary producer

Decomposer
720 Chapter 29 Microbial Ecology

(oxidize inorganic chemicals for energy). Primary produc-

ers serve as a food source for consumers and decomposers.
photoautotroph, p. 93 chemolithoautotroph, p. 93

■ Consumers. These are heterotrophs that eat primary produc-

ers or other consumers. Herbivores, which eat plants or algae,
are primary consumers. Carnivores that eat herbivores are
secondary consumers; carnivores that eat other carnivores are
tertiary consumers. A chain of consumption is a food chain;
interacting food chains are a food web. Growth Transfers
■ Decomposers. These are heterotrophs that digest the remains and more
growth
of primary producers and consumers. The fresh or partially
decomposed organic matter used as a food source—including
carcasses, excreta, and plant litter—is called detritus.
Decomposers specialize in digesting complex materials such
as cellulose, converting them into small molecules that can
more easily be used by other organisms. The complete break-
down of organic molecules into inorganic molecules such as
ammonia, sulfates, phosphates, and carbon dioxide is called
mineralization. Microorganisms, particularly bacteria and
fungi, play a major role in decomposition because of their
ubiquity and unique metabolic capabilities. Coccus Rod
FIGURE 29.2 Competition The bacterium that multiplies faster
yields the larger population.
Microbes in Low-Nutrient Environments ? Why does transferring some of the culture make it easier to see
Low-nutrient environments such as lakes, rivers, and streams are the effects of competition?
common in nature, so microorganisms that can grow in dilute
aqueous solutions are widespread. Most microbial growth in these Antagonism among groups of organisms also helps deter-
settings is in biofilms, and the cells are shed from the film into the mine the makeup of a community. In the soil, for example, some
aqueous solution. microbes resort to a type of chemical warfare, producing antimi-
In addition to natural low-nutrient environments, microorgan- crobial compounds. Bacteriocins, proteins produced by bacteria
isms even grow in distilled-water reservoirs such as those found that kill closely related strains, are an example of antagonistic
in research laboratories and pulmonary mist therapy units used chemicals that play an important role in microbial ecosystems,
in hospitals. The microbes extract the trace amounts of nutrients promoting biodiversity through competition. It is tempting to
absorbed by the water from the air or adsorbed onto the biofilm. speculate that antibiotics produced by Streptomyces species share
Although the organisms grow slowly, they can reach concentra- a similar function, but their natural role is still poorly understood.
tions as high as 107 per milliliter. This cell concentration is not Recent evidence suggests that they play an important role in cell
high enough to result in a cloudy solution, so the growth usually to cell signaling. the genus Streptomyces, p. 268
goes unnoticed. This can have serious consequences for the health
of hospitalized patients and for the success of laboratory experi-
ments that depend on water purity.
Microorganisms and
Organisms that grow in dilute environments contain highly Environmental Changes
efficient transport systems for moving nutrients inside the cell. Environmental changes often result in alterations in a community.
Other mechanisms that bacteria use to thrive in dilute aquatic Those organisms that have adapted to live several inches beneath
environments are described in chapter 11. transport systems, p. 55 the surface of an untilled field will probably not be well suited to
thriving in aquatic environments, p. 269 growth in that field if it is plowed, fertilized, and irrigated. In addi-
tion to external sources of environmental change, the growth and
metabolism of organisms themselves can alter the environment
Microbial Competition and Antagonism dramatically. Nutrients may become depleted, and a variety of
Perhaps nowhere in the living world is competition more fierce waste products, many of which are toxic, may accumulate.
and the results of competition more quickly evident than among In some environments, the changing conditions bring about
microorganisms. The ability of an organism to compete success- a highly ordered and predictable succession of bacterial species.
fully for a habitat is generally related to the rate at which the An example of this occurs in unpasteurized milk, which usually
organism multiplies, as well as to its ability to withstand adverse contains various species of microbes derived mainly from the
environmental conditions. Because bacteria multiply logarithmi- immediate environment around the cow. Initially, the dominant
cally, any small differences in their generation times will result in bacterium is Lactococcus lactis, which breaks down the milk
a very large difference in the total number of cells of each species sugar lactose, forming lactic acid as a fermentation end product
after a relatively short time (figure 29.2). (figure  29.3). This sours the milk and also denatures the milk
 Part V Applied Microbiology 721

Most
acidic
Acidity
(pH)
6.8 4.0 3.0 4.0 5.5 6.8
Lactococcus Lactobacillus sp. Yeasts and Putrefying
lactis molds bacteria
Number of organisms

0 7 14
Time (days)

FIGURE 29.3 Growth of Microbial Populations in

Unpasteurized Raw Milk at Room Temperature Production
of acid causes souring and encourages growth of fungi. Eventually,
bacteria digest the proteins, causing putrefaction.
? Why does the pH of the milk first decrease and then increase?
FIGURE 29.4 A Microbial Mat A microbial mat is a thick, dense,
highly organized structure composed of distinct layers of different
proteins, causing the milk to curdle. The acid inhibits most other groups of microorganisms.
organisms in the milk, and eventually enough acid is produced
to inhibit L. lactis. Lactobacillus species can multiply in this ? Why are the middle layers of the mat reddish-pink? Why are the
lower layers black?
highly acidic environment, however, and these bacteria metabo-
lize any remaining sugar, forming more acid until their growth is
also inhibited. Yeasts and molds, which tolerate even more acid produced by the photosynthetic bacteria growing in the mat’s
conditions, then become the dominant group. They oxidize the upper layers, using sulfate as a terminal electron acceptor.
lactic acid, which raises the pH. Most of the sugar has already cyanobacteria, p.  261 photosynthetic pigments, p.  152 purple sulfur
been used at this point, so the streptococci and lactobacilli can- bacteria, p. 259 sulfate-reducers, p. 258 terminal electron acceptor, p. 130
not resume multiplication. Milk protein (casein) is still available, Although microbial mats can be found in a variety of areas,
however, and can be used by protease-producing members of the those near hot springs in Yellowstone National Park are some of
endospore-forming genus Bacillus. This breakdown of protein, the most intensively studied. The mats in these extreme areas are
known as putrefaction, yields a completely clear and foul-smelling undisturbed by grazing eukaryotic organisms and, consequently,
product. The milk thus goes through a succession of changes with provide an important model for studying microbial interactions.
time, first souring and finally putrefying. proteases, p. 150

Studying Microbial Ecology

Microbial Communities Because so few microorganisms can be successfully cultivated in
Microorganisms most often grow as biofilms attached to solid the laboratory, studying only those that have been isolated often
surfaces or at air-water interfaces. General aspects of biofilms does not give an accurate picture of what actually occurs in nature.
were described in detail in chapter 4. In this section, we focus on Molecular techniques are now complementing the traditional
a specific type of biofilm—a microbial mat. biofilms, p. 84 methods such as culture and microscopy, allowing researchers to
A microbial mat is a thick, dense, highly organized structure better understand complex microbial communities.
composed of distinct layers. Frequently they are green, reddish- Microscopic methods can now be used to examine the com-
pink, and black, which indicate the growth of different microbial position of microbial populations. For example, certain dyes
groups (figure  29.4). The top green layer is typically composed are made fluorescent by metabolic activities carried out only
of various species of cyanobacteria, and the color is due to their in living cells, and therefore can be used to selectively observe
photosynthetic pigments. Directly below the green layer is a viable cells (see figure 3.19a). A different technique, fluorescence
reddish-pink layer consisting of purple sulfur bacteria. The light- in situ hybridization (FISH), uses nucleic acid probes labeled
harvesting pigments of these anoxygenic phototrophs can use with a fluorescent molecule to observe only cells that contain
wavelengths of light not collected by the cyanobacteria. At the specific nucleotide sequences (see figure  9.20). Scanning laser
bottom is a black layer, resulting from iron molecules reacting with microscopes allow researchers to observe sectional views of a
hydrogen sulfide produced by a group of bacteria called sulfate- three-dimensional specimen such as a biofilm (see figure  3.8).
reducers. These obligate anaerobes oxidize the organic compounds fluorescent dyes, p. 49 FISH, p. 232 scanning laser microscopes, p. 44
722 Chapter 29 Microbial Ecology

Polymerase chain reaction (PCR) can be used to detect cer- Marine environments such as the oceans cover more than 70%
tain organisms and assess population characteristics. To detect of the earth’s surface. They are the most abundant aquatic habi-
a specific organism, primers are selected that amplify DNA tat, representing about 95% of the global water. The freshwater
unique to that organism (see figure  9.14). To study the compo- environments—lakes and rivers—represent only a small fraction
sition of a population, total 16S rRNA gene segments can be of the total water.
amplified. Individual fragments can then be cloned and studied. Deep lakes and oceans have characteristic zones that influ-
Alternatively, the set of amplified sequences can be separated ence the distribution of microbial populations. The uppermost
and examined using a technique called denaturing gradient gel layer—where sufficient light penetrates—supports the growth of
electrophoresis (DGGE). This procedure gradually denatures photosynthetic microorganisms, including algae and cyanobacte-
double-stranded nucleic acid during gel electrophoresis and, as ria. The organic material synthesized by these primary producers
a consequence, separates fragments of similar size according to gradually descends and is then metabolized by heterotrophs.
their melting point, which is related to the nucleotide sequence. The number of microorganisms in waters is influenced by the
Using DGGE, a mixture of 16S rRNA fragments with different nutrient content. In oligotrophic waters, meaning nutrient poor,
sequences will resolve into a distinct pattern of bands. PCR and the growth of photosynthetic organisms and other autotrophs is
DDGE studies have confirmed that standard culture techniques limited by the lack of inorganic nutrients, particularly phosphate,
can be poor indicators of the composition of natural microbial nitrate, and iron. When waters are eutrophic (nutrient rich), pho-
populations. The molecular techniques, which show the relative tosynthetic organisms flourish, often forming a visible layer on
abundance of specific nucleotide sequences, confirm that species the surface (figure 29.5). In turn, photosynthesis produces organic
predominating in laboratory culture often represent only a frac- compounds that foster the growth of heterotrophs in lower layers.
tion of the total population. polymerase chain reaction, p. 227 gel The heterotrophs consume dissolved O2 as they metabolize the
electrophoresis, p. 217 organic material. Because O2 consumption can outpace the slow
Genomics is also advancing the study of microbial ecology rate of diffusion of atmospheric O2 into the waters, the environ-
because sequence information gleaned from one species can be ment can become hypoxic (very low in dissolved O2). Insufficient
applied to others. For example, researchers found that varia- O2 leads to the death of resident fish and other aquatic animals.
tions of a gene coding for bacterial rhodopsin, a light-sensitive
pigment that provides a mechanism for harvesting the energy of Marine Environments
sunlight, are widespread in marine bacteria. This gene provides
bacteria with a mechanism for phototrophy that does not require Marine environments range from the deep sea, where nutrients
chlorophyll and might be an important mechanism for energy are scarce, to the shallower coastal regions, where nutrients may
accumulation in ocean environments. genomics, p. 184 bacterial
rhodopsin, p. 278
Scientists are also using metagenomics—the study of total
genomes in a sample—to study microbial populations in their
natural environment. metagenomics, p. 184

MicroAssessment 29.1
Microorganisms play a major role in most ecosystems. Organisms
are categorized as primary producers, consumers, or decomposers.
Competition among microorganisms in a habitat can be intense.
Microbial mats have distinct layers. Molecular techniques are
allowing researchers to better understand natural microbial
communities.
1. What are the roles of primary producers, consumers, and
decomposers?
2. Why are molecular techniques important in studying microbial
ecology?
3. How could FISH (fluorescence in situ hybridization) be used to
determine the relative proportions of archaea and bacteria in a
population? +

29.2 ■ Aquatic Habitats

FIGURE 29.5 Eutrophication in a Polluted Stream Photo-
Learning Outcome synthetic organisms flourish in the nutrient-rich water. The organic
compounds they produce permit abundant growth of heterotrophs in
7. Compare and contrast the habitats provided by marine, freshwater, lower layers.
and specialized aquatic environments.
? How would the cows in the photo contribute to eutrophication?
 Part V Applied Microbiology 723

be abundant due to runoff from the land. Seawater contains in the process. This causes a large region in the Gulf—sometimes
about 3.5% salt, compared with about 0.05% for fresh water. in excess of 7,000 square miles—to become hypoxic. Animals in
Consequently, it supports the growth of halophilic organisms, the area either flee or die. Nutrient enrichment of coastal waters
which prefer or require high salt concentrations, and halotolerant also contributes to blooms of toxin-producing algae. medical
ones. Temperatures often vary widely at the surface, but decrease importance of algae, p. 291
with depth until reaching about 2°C in the deeper waters; an
exception is the areas around hydrothermal vents, which will be
described later. halophilic, p. 92 hydrothermal vents, p. 729
Freshwater Environments
Ocean waters are typically oligotrophic, limiting the growth As with marine environments, the types and relative numbers
of microorganisms. The small amounts of organic material pro- of microbes inhabiting fresh waters depend on multiple factors
duced by photosynthetic organisms is quickly consumed as it including light, concentration of dissolved O2 and nutrients, and
descends, so few nutrients reach the sediments below. Even in the temperature.
deep sea, marine water is O2-saturated due to mixing associated Oligotrophic lakes in temperate climates may have anaerobic
with tides, currents, and wind action. layers due to thermal stratification resulting from seasonal tem-
The ecology of inshore areas is not as stable as the deep perature changes. During the summer months, the surface water
sea and can be dramatically affected by nutrient-rich runoff. An warms. This decreases the density of the water, causing it to form
unfortunate example is the dead zone—a region devoid of fish a distinct layer that does not mix with the cooler, denser water
and other marine life—that forms in the Gulf of Mexico, as well below. The upper layer, called the epilimnion, is generally O2 rich
as other areas (figure  29.6). The Mississippi River, carrying due to the activities of photosynthetic organisms. In contrast, the
nutrients accumulated as it runs through agricultural, industrial, lower layer, the hypolimnion, may be anaerobic due to the con-
and urbanized regions, feeds into the Gulf. As a consequence of sumption of O2 by heterotrophs. Separating these two layers is the
this nutrient-enrichment, populations of algae and cyanobacteria thermocline, a zone of rapid temperature change. As the weather
flourish in the spring and summer when sunlight is also plentiful. cools, the waters mix, providing O2 to the deep water.
Heterotrophic microbes then metabolize the organic compounds Rapidly moving waters, such as rivers and streams, are very
synthesized by these primary producers, consuming dissolved O2 different from lakes. They are usually shallow and turbulent,
facilitating O2 circulation, so they are generally aerobic. Light
may penetrate to their bottom, making photosynthesis possible.
Algae and cyanobacteria flourish, Sheathed bacteria such as Sphaerotilus sp. and Leptothrix sp.
using photosynthesis to produce commonly adhere to rocks and other solid structures, where they
organic compounds.
then use nutrients that flow by. sheathed bacteria, p. 269
Nutrient-rich water
Specialized Aquatic Environments
Specialized aquatic environments include salt lakes, such as the
Great Salt Lake in Utah, which have no outlets. As water in these
lakes evaporates, the salt concentrations become much higher than
that in seawater. Extreme halophiles thrive in this environment.
Organic extreme halophiles, p. 92
compounds
Other specialized habitats include iron springs that contain
large quantities of ferrous ions; these springs are habitats for spe-
Aerobic decomposition of organic
compounds by heterotrophic cies of Gallionella and Sphaerotilus. Sulfur springs support the
microbes depletes O 2. growth of both photosynthetic and non-photosynthetic sulfur bac-
teria. Other aquatic environments include groundwater, stagnant
ponds, swimming pools, and drainage ditches, each offering its
own opportunity for bacterial growth.

MicroAssessment 29.2
Aquatic habitats include marine, freshwater, and specialized
environments. Nutrient-rich waters can become hypoxic. Lakes often
Animals flee hypoxic
environment or die.
exhibit thermal stratification during summer months.
4. Compare the salt content of seawater with that of fresh water
and salt lakes.
5. Explain how nutrient-rich runoff can cause waters to become
hypoxic.
FIGURE 29.6 Dead Zone Formation 6. Why would the nutrient content of a body of water be more
homogeneous than that of a terrestrial environment? +
? Why does decomposition by microbes deplete O2?
724 Chapter 29 Microbial Ecology

29.3 ■ Terrestrial Habitats Microorganisms in Soil

The density and composition of the soil microbiota are dramati-
Learning Outcome cally affected by environmental conditions. Wet soils, for exam-
8. Describe soil as a microbial habitat. ple, are unfavorable for aerobic microbes because the spaces in
the soil fill up with water, diminishing the amount of air in the
Although microbes can adhere to and grow on a variety of objects soil. When the water content of soil drops to a very low level, as
on land, the focus in this section is soil—a critical component during a drought or in a desert environment, the metabolic activ-
of terrestrial ecosystems. Extreme terrestrial habitats, such as ity and number of soil microorganisms decrease. Many species
volcanic vents and fissures, and some of the extremophiles that of soil organisms produce survival forms such as endospores and
inhabit them are described in chapter 11. archaea that thrive in cysts that are resistant to drying. Other environmental influences
extreme conditions, p. 278 that affect soil microbes include acidity, temperature, and nutri-
One reason scientists are interested in soil is that the ent supply. For example, acidity suppresses bacterial growth,
microbes there synthesize a variety of useful chemicals. For allowing fungi to thrive with less competition for nutrients. This
example, the various species of Streptomyces produce over 500 is why mushrooms often appear in a lawn fertilized with an acid-
different antibiotic substances, at least 50 of which have useful producing fertilizer such as ammonium chloride.
applications in medicine, agriculture, and industry. The phar- Prokaryotes are the most numerous soil inhabitants. Their
maceutical industry has tested many thousands of soil micro- physiological diversity allows them to colonize all types of soil.
organisms in search of those that produce useful antibiotics. In In general, Gram-positive bacteria are more abundant in soils
addition, soil microbes are being investigated for their ability to than Gram-negative bacteria. Among the most common Gram-
degrade toxic chemicals, an application of microbiology called positive bacteria are members of the genus Bacillus. These form
bioremediation, which will be discussed in chapter 30. Probably endospores, allowing them to survive long periods of adverse
in no other habitat can one find a greater range of biosynthetic conditions such as drought or extreme heat. Streptomyces spe-
and biodegradative capabilities than are represented in the soil. cies produce conidia, which are desiccation-resistant structures.
bioremediation, p. 744 They also produce metabolites called geosmins, which give soil
its characteristic musty odor. As discussed earlier, Streptomyces
species produce many medically useful antibiotics. Other bacteria
Characteristics of Soil adapted to thrive in terrestrial environments—including myxobac-
Soil is composed of pulverized rock, decaying organic material, teria and species of Clostridium, Azotobacter, Agrobacterium, and
air, and water. It teems with life, including bacteria, fungi, algae, Rhizobium—were discussed in chapter 11. endospore-formers, p. 67
protozoa, worms, insects, and plant roots. Soil communities may the genus Streptomyces, p. 268 thriving in terrestrial environments, p. 266
contain more than 4,000 different species per gram of soil. The Although prokaryotes are the most numerous soil microbes,
top 6 inches of fertile soil may harbor more than 2 tons of bacteria the biomass of fungi is much greater. Most fungi are aerobes, so
and fungi per acre! Soil represents an environment that can change they usually grow in the top 10 cm of soil. The soil fungi degrade
abruptly and dramatically. Heavy rains, for example, can cause a complex macromolecules such as lignin (the major component
soil to rapidly become waterlogged. Trees dropping their leaves of cell walls of woody plants) and cellulose. Some soil fungi are
can suddenly enrich the soil with organic nutrients. Farmers and free-living, and others live in symbiotic relationships. The latter
gardeners rapidly change the nutrient mix by applying fertilizers. include mycorrhizas—fungi growing in a symbiotic relationship
Soil forms as rock weathers. Water, temperature changes, with certain plant roots. mycorrhizas, p. 286
windblown particles, and other physical forces gradually cause In addition to bacteria and fungi, various algae and protozoa
the rock to crack and break. Photosynthetic organisms includ- are found in most soils. Algae depend on sunlight for energy,
ing algae, mosses, and lichens growing on the surfaces of rocks so they mostly live on or near the soil surface. Most protozoa
synthesize organic compounds. Various bacteria and fungi then require O2, so they too are found near the surface, typically where
use these compounds as carbon and energy sources, producing microbes on which they feed are plentiful.
acids and other chemicals that gradually decompose the rocks.
As soil slowly forms, some plants begin to grow. When these die
and decay, the residual organic material functions as a sponge, The Rhizosphere
retaining water and thus allowing more plants to grow. Over time, The concentration of microbes, particularly Gram-negative bac-
more organic compounds accumulate, forming a slowly degrading teria, is generally much greater in the rhizosphere (the zone of
complex polymeric substance called humus. lichens, p. 286 soil that adheres to plant roots) than in surrounding soil. This is
The texture of the soil influences its porosity, which in turn because the root cells secrete organic molecules, enriching the
impacts the amount of air exchange and how much water can flow region. Particular bacterial species appear to preferentially inter-
through. Finely textured soils, such as clay soils, are more likely act with certain plants. For example, the rhizosphere of certain
to become waterlogged and anaerobic. In contrast, sandy soils that grasses can have high concentrations of Azospirillum species,
dry quickly allow water to pass through and are generally aerobic. which fix nitrogen. nitrogen fixation, p. 726
 Part V Applied Microbiology 725

MicroAssessment 29.3 in the atmosphere raises global temperatures because the gases
absorb infrared radiation and reflect it back to earth.
The density and composition of the soil are dramatically affected When studying biogeochemical cycles, it is helpful to bear
by environmental conditions. The concentration of microbes in the
rhizosphere is generally much higher than that of the surrounding soil.
in mind the role of a given element in a particular organism’s
metabolism. Elements are used for three general purposes:
7. Why are wet soils unfavorable for aerobic organisms?
8. What is the significance of the rhizosphere? ■ Biosynthesis (biomass production). All organisms require
9. How can the biomass of fungi in soil be greater considering that elements for biosynthesis. As an example, nitrogen is required
bacteria are more numerous? + to produce amino acids. Plants and many prokaryotes assimi-
late nitrogen by incorporating ammonia (NH3) to synthesize
the amino acid glutamate (see figure  6.29a). Some prepare
for this step by converting nitrate (NO3–) to ammonia. Once
29.4 ■ Biogeochemical Cycling glutamate has been synthesized, the amino group can then be
and Energy Flow transferred to other carbon compounds to produce the neces-
sary amino acids. Animals cannot incorporate ammonia and
Learning Outcomes instead require amino acids in their diet. Some prokaryotes
9. Diagram the carbon, nitrogen, sulfur, and phosphorus cycles, and can reduce atmospheric nitrogen to form ammonia—the
describe some of the important microbial contributors. process of nitrogen fixation. The ammonia can then be incor-
10. Compare and contrast energy cycling in environments with porated into cellular material. amino acid synthesis, p. 156
sunlight versus those far removed from sunlight. ■ Energy source. Reduced carbon compounds such as sugars,
lipids, and amino acids are used as energy sources by chemo-
Biogeochemical cycles are the cyclical paths that elements take organotrophs. Chemolithotrophs can use reduced inorganic
as they flow through living (biotic) and non-living (abiotic) molecules such as hydrogen sulfide (H2S), ammonia (NH3),
components of ecosystems. These cycles are important because a and hydrogen gas (H2) (see table 4.5). energy source, p. 130
fixed and limited amount of the elements that make up living cells ■ Terminal electron acceptor. In aerobic conditions, O2 is
exists on the earth and in the atmosphere. Thus, in order for an
used as a terminal electron acceptor. In anaerobic conditions,
ecosystem to sustain its characteristic life-forms, elements must
some prokaryotes can use nitrate (NO3–), nitrite (NO2–), sul-
continuously be recycled. For example, the organic carbon that
fate (SO4–), or carbon dioxide (CO2) as a terminal electron
animals use as an energy source is then exhaled as carbon dioxide
acceptor. terminal electron acceptor, p. 130
(CO2); if this inorganic form of carbon were not eventually con-
verted back to an organic form, we would run out of the organic
carbon needed for growth. The carbon and nitrogen cycles are Carbon Cycle
particularly important because they involve stable gaseous forms
(carbon dioxide and nitrogen gas), which enter the atmosphere and All organisms are composed of organic molecules such as pro-
thus have global impacts. teins, lipids, and carbohydrates. The carbon travels through the
Although elements continually cycle in an ecosystem, energy food chain as primary producers are eaten by primary consumers,
does not. Instead, energy must be continually added to an ecosys- which are then eaten by secondary consumers. Decomposers then
tem, fueling the activities required for life. use the remains of primary producers and consumers.
Understanding the cycling of nutrients and the flow of energy
is particularly important as human activities affect the environ- Carbon Fixation
ment in a major way. For example, industrial processes that A fundamental aspect of the carbon cycle is carbon fixation, the
convert nitrogen gas (N2) into ammonia-containing fertilizers defining characteristic of primary producers (figure 29.7). These
have boosted food production substantially, but they also alter the organisms all convert CO2 into an organic form, using mecha-
nitrogen cycle by increasing the amount of fixed nitrogen avail- nisms described in chapter 6. Without primary producers, no other
able. When these nitrogen sources pollute lakes and coastal areas, organisms, including humans, could exist. We depend on them
eutrophication can result. As a consequence, the dissolved O2 is to generate the organic carbon compounds we use as an energy
depleted, leading to the death of aquatic animals. The extra nutri- source and for biosynthesis. carbon fixation, p. 154
ents also lead to decreased biodiversity in terrestrial ecosystems.
Excavation and burning of coal, oil, and other carbon-rich fossil Respiration and Fermentation
fuels provides energy for our daily activities, but releases addi- When heterotrophs consume organic material, they break it down
tional CO2 and other carbon-containing gases into the atmosphere. using respiration and/or fermentation to release the energy, which
Fossil fuels, the ancient remains of partially decomposed plants is captured to make ATP. The processes usually make CO2.
and animals, are nutrient reservoirs unavailable without human The type of organic material helps dictate which species
intervention, and therefore would not normally participate in degrade it. A wide variety of organisms use sugars, amino acids, and
biogeochemical cycles. The increase of carbon-containing gases proteins as energy sources, but rapidly multiplying bacteria often
726 Chapter 29 Microbial Ecology

Organic
compounds The O2 supply has a strong influence on the carbon
Aerobic cycle. Not only does O2 allow degradation of certain com-
pounds such as lignin, it also helps determine the types of
carbon-containing gases produced. When organic matter is
Carbon fixation Aerobic respiration degraded aerobically, a great deal of CO2 is produced. When
Reduction of CO 2 Oxidation of organic the O2 level is low, however, as is the case in marshes,
as a carbon source compounds as an swamps, and manure piles, the degradation is incomplete,
for biosynthesis energy source
generating some CO2 and a variety of other products.
Methane oxidation
Oxidation of
Methanogenesis and Methane Oxidation
methane as an In anaerobic environments, CO2 is used by methanogens.
energy source These archaea obtain energy by oxidizing hydrogen gas,
using CO2 as a terminal electron acceptor, generating meth-
CH 4 CO 2
ane (CH4). Methane that enters the atmosphere is oxidized
Methanogenesis
Reduction of CO 2
by ultraviolet light and chemical ions, forming carbon mon-
as a terminal oxide (CO) and CO2. A group of microorganisms called
electron acceptor methylotrophs can use methane as an energy source, oxidiz-
ing it to produce CO2. methanogens, p. 256

Anaerobic respiration Carbon fixation

and fermentation
Oxidation of organic compounds
Reduction of CO 2
as a carbon source
Nitrogen Cycle
as an energy source; fermentation for biosynthesis Nitrogen is a component of proteins and nucleic acids. As
uses organic compounds as a
terminal electron acceptor as well consumers ingest plants and animals for carbon and energy,
Anaerobic they also obtain their required nitrogen. Prokaryotes, as
a group, are far more diverse in their use of nitrogen-
Organic containing compounds. Some use oxidized nitrogen com-
compounds pounds such as nitrate (NO3–) and nitrite (NO2–) as terminal
electron acceptors; others use reduced nitrogen compounds
FIGURE 29.7 Carbon Cycle Blue arrows represent steps where carbon
compounds are used as energy sources; red arrows represent steps where such as ammonium (NH4+) as energy sources. These meta-
carbon compounds are used as terminal electron acceptors, and green bolic activities represent essential steps in the nitrogen cycle
arrows represent steps where inorganic carbon is used in biosynthesis. (figure 29.9).
? What is the role of autotrophs in the carbon cycle?
Nitrogen Fixation
play the dominant role in decomposing these substances. In contrast, Nitrogen fixation is the process in which nitrogen gas (N2) is
only certain fungi can break down lignin, a major component of wood reduced to form ammonia (NH3), which can then be incorporated
(figure 29.8). Aerobic conditions are required for this degrada- into cellular material. The process, catalyzed by the enzyme com-
tion, so wood at the bottom of marshes resists decay. plex nitrogenase, requires a tremendous amount of energy because
N2 has a very stable triple bond.
Although the atmosphere consists of approximately 80%
N2, relatively few organisms—all of which are prokaryotes—can
reduce this gaseous form of the element. Thus, just as humans and
other animals depend on primary producers to fix carbon, they
rely on prokaryotes to convert atmospheric nitrogen to a form
they can assimilate to create biomass.
Some nitrogen-fixing prokaryotes, or diazotrophs, are free-
living, whereas others form symbiotic associations with higher
organisms, particularly certain plants. Among the free-living
examples are members of the genus Azotobacter. These hetero-
trophic, aerobic, Gram-negative rods may be the chief suppliers
of fixed nitrogen in ecosystems such as grasslands that lack plants
with nitrogen-fixing symbionts. The dominant free-living, anaero-
bic, soil diazotrophs are certain members of the genus Clostridium.
Certain cyanobacteria are diazotrophs; these photosynthetic bacte-
FIGURE 29.8 Wood-Degrading Fungus Growing on a Dead ria use both nitrogen and carbon from the atmosphere. Symbiotic
Tree These fungi thrive in wet conditions and digest lignin, the diazotrophs will be discussed later in the chapter. symbiotic
major cell wall component of woody plants. nitrogen-fixers and plants, p. 730 the genus Azotobacter, p. 267 the genus
? Dead trees submerged in marshes resist decay. Why? Clostridium, p. 258 nitrogen-fixing cyanobacteria, p. 262
 Part V Applied Microbiology 727

NO2-
(nitrite)
are then transported into the cell, and the amino groups
removed, releasing ammonium. The decomposer will assim-
Nitrification ilate much of this to create biomass. Some will be released
Oxidation of NH4+ into the environment, however, where it can be assimilated
and NO2- as by plants and other organisms. deamination, p. 150
energy sources

Nitrification
Organic Aerobic Nitrification is the process that oxidizes ammonium (NH4+)
Ammonium Nitrate
assimilation nitrogen assimilation to nitrate (NO3–). A group of bacteria known collectively as
Amination of Reduction of NO3- nitrifiers do this in a cooperative two-step process, using
organic compounds as a nitrogen source ammonium and an intermediate, nitrite (NO2–), as energy
for biosynthesis for biosynthesis
sources. Nitrifiers are obligate aerobes, using O2 as a termi-
nal electron acceptor. Consequently, nitrification does not
NH4+ NO3- occur in waterlogged soils or in anaerobic regions of aquatic
(ammonium) Ammonification (nitrate) environments. nitrifiers, p. 727
Deamination of
organic compounds Nitrification has some important consequences with
so they can be used respect to agricultural practices and pollution. Farmers often
for catabolism apply ammonium-containing compounds to soils as a source
Nitrogen of nitrogen for plants. The ammonium is retained by soils,
fixation because its positive charge causes it to adhere to negatively
Reduction of N2 Anammox Anaerobic charged soil particles. Nitrification converts the ammonium
to ammonia as a Anaerobic oxidation
nitrogen source of NH4+ as an
to nitrate, a form of nitrogen more readily used by plants, but
for biosynthesis energy source rapidly leached from soil by rainwater. To slow nitrification,
NO2-
(nitrite) certain chemicals can be added to ammonium-fertilized soils.
N2
(nitrogen gas) Denitrification Leaching of nitrate and nitrite from soil is a health con-
Reduction of NO3- cern if the compounds contaminate drinking water. Nitrite is
or NO2- as a terminal toxic because it can combine with hemoglobin of the blood,
electron acceptor
reducing blood’s O2-carrying capacity. Even nitrate, which
FIGURE 29.9 Nitrogen Cycle Blue arrows represent steps where in itself is not very toxic, can be dangerous if high levels
nitrogen compounds are used as energy sources; red arrows represent are ingested because some intestinal bacteria can use it as a
steps where nitrogen compounds are used as terminal electron acceptors,
and green arrows represent steps where inorganic nitrogen is used in
terminal electron acceptor, converting it to nitrite.
biosynthesis. Not all parts of the cycle are shown.
? Why are prokaryotes critical in the nitrogen cycle?
Denitrification
Denitrification is the process that reduces nitrate (NO3–),
converting it to gaseous forms such as nitrous oxide (N2O)
Energy-expensive chemical processes can fix nitrogen to
and molecular nitrogen (N2). This happens when prokaryotes
make fertilizers, and these are playing an increasingly larger
anaerobically respire using nitrate as a terminal electron acceptor.
role in the nitrogen cycle. In fact, fixed nitrogen sources associ-
anaerobic respiration, p. 134
ated with human intervention, including fertilizer production and
Denitrification can have negative environmental and eco-
planting crops that foster the growth of symbiotic nitrogen-fixers,
nomic consequences. Under anaerobic conditions in wet soils, for
now appear to surpass natural biological nitrogen fixation.
example, denitrifying bacteria will reduce the oxidized nitrogen
MicroByte compounds of fertilizers, releasing gaseous nitrogen to the atmo-
Nitrogenase uses approximately 16 molecules of ATP for every sphere. In some areas, this process may represent 80% of nitro-
molecule of nitrogen fixed. gen lost from fertilized soil, a considerable economic loss to the
farmer. In addition, nitrous oxide contributes to global warming.
Denitrification is not always undesirable. The process can
Ammonification be actively fostered in certain steps of wastewater treatment as a
Ammonification is the decomposition process that converts means to remove nitrate. This compound could otherwise act as
organic nitrogen into ammonia (NH3). In alkaline environments, a fertilizer in the waters to which the wastewater is discharged,
such as heavily limed soil, the gaseous ammonia may enter promoting algal growth. microbiology of wastewater treatment, p. 736
the atmosphere. In neutral environments, ammonium (NH4+) is
formed. This positively charged ion adheres to negatively charged Anammox
particles. Certain bacteria oxidize ammonium under anaerobic conditions,
Proteins, which are among the most common nitrogen- using nitrite as a terminal electron acceptor. This reaction, called
containing organic compounds, can be degraded by a wide variety anammox (for anoxic ammonia oxidation), forms N2 and might
of microbes. The microbes secrete proteolytic enzymes that break provide an economical means of removing nitrogen compounds
down proteins into short peptides or amino acids. These products during wastewater treatment.
728 Chapter 29 Microbial Ecology

Sulfur Cycle species and the largest known bacterium, Thiomargarita

namibiensis, have unusual mechanisms to cope with the fact
Sulfur is found in all living matter, chiefly as a component of the
that their energy source and terminal electron acceptor are found
amino acids methionine and cysteine. Like the nitrogen cycle, key
in two different environments. sulfur-oxidizing bacteria, p.  262
steps of the sulfur cycle depend on the activities of prokaryotes
sulfur-oxidizing, nitrate-reducing marine bacteria, p. 273
(figure 29.10).
Hydrogen sulfide and elemental sulfur are oxidized anaerobi-
cally by photosynthetic green and purple sulfur bacteria. These
Sulfur Assimilation and Decomposition
bacteria harvest energy from sunlight but require reduced mole-
Most plants and microorganisms assimilate sulfur as sulfate cules as a source of electrons to generate reducing power. Like the
(SO42-), reducing it to form biomass. Like nitrogen, organic sul- chemolithotrophs that use hydrogen sulfide and elemental sulfur,
fur is present chiefly as a part of the amino acids that make up the photosynthetic sulfur oxidizers produce sulfate. green sulfur
proteins. Decomposition of the sulfur-containing amino acids bacteria, p. 260 purple sulfur bacteria, p. 259
releases hydrogen sulfide (H2S), a gas.
Sulfur Reduction
Sulfur Oxidation
Under anaerobic conditions, sulfate generated by the sulfur-
Hydrogen sulfide (H2S) and elemental sulfur (S0) can both oxidizers can then be used as a terminal electron acceptor by
serve as an energy source for certain chemolithotrophs. Sulfur- certain organisms. The sulfur- and sulfate-reducing bacteria and
oxidizing prokaryotes, including species of Beggiatoa, Thiothrix, archaea use sulfate in the process of anaerobic respiration, reduc-
and Thiobacillus, oxidize these molecules to sulfate (SO42–). ing it to hydrogen sulfide (H2S). In addition to its unpleasant odor,
Certain prokaryotes in anaerobic marine environments can oxidize the H2S is a problem because it reacts with metals, resulting in
elemental sulfur, using nitrate as a terminal electron acceptor. As corrosion. sulfur- and sulfate-reducing bacteria, p. 258
discussed in chapter 11, these organisms, including Thioploca

Phosphorus Cycle and Other Cycles

S0
(elemental sulfur) Phosphorus is a component of several critical biological com-
pounds including nucleic acids, phospholipids, and ATP. Most
Sulfur oxidation
Oxidation of H2S and plants and microorganisms can take up phosphorus as ortho-
Aerobic
elemental sulfur as phosphate (PO43–), incorporating it into biomass. From there,
energy sources the phosphorus is passed along the food web. When plants and
animals die, decomposers convert organic phosphate back to the
inorganic form.
In many aquatic habitats, growth of algae and cyanobacteria—
H2S the primary producers—is limited by low concentrations of
(hydrogen Organic sulfur SO42- phosphorus. When phosphates are added from sources such as
sulfide) (sulfate) agricultural runoff, phosphate-containing detergents, and waste-
Decomposition Sulfate
water, eutrophication can result.
Release of H2S from assimilation Other important elements, including iron, calcium, zinc,
organic compounds so Reduction of SO42- manganese, cobalt, and mercury, are also recycled by microor-
they can be used for as a sulfur source
catabolism for biosynthesis
ganisms. Many prokaryotes contain plasmids coding for enzymes
that carry out oxidation of metallic ions.
Anaerobic

Energy Sources for Ecosystems

S0 All chemotrophs harvest the energy trapped in chemical bonds to
(elemental sulfur) generate ATP. This energy cannot be totally recycled, however,
Sulfate reduction because a portion is always lost as heat when bonds are broken.
Reduction of SO42- Thus, energy is continually lost from biological systems. To com-
or elemental sulfur pensate, energy must be added to ecosystems.
as a terminal electron
acceptor Photosynthesis, carried out by chlorophyll-containing plants
and microorganisms, converts radiant energy (sunlight) to chemi-
FIGURE 29.10 Sulfur Cycle Blue arrows represent steps where
sulfur compounds are used as energy sources; red arrows represent
cal energy in the form of organic compounds, which can be used
steps where sulfur compounds are used as terminal electron by chemoorganotrophs. The requirement for radiant energy has
acceptors, and green arrows represent steps where inorganic sulfur traditionally been used to explain why life is not equally abundant
is used in biosynthesis. everywhere. However, the discovery of different types of com-
? Organisms that oxidize sulfur use it for what purpose in their munities far removed from sunlight, including near hydrothermal
metabolism? vents and within rocks, has dramatically altered this idea. These
 Part V Applied Microbiology 729

communities rely on chemolithoautotrophs, which harvest the FIGURE 29.11

energy of reduced inorganic compounds and use it to form organic Hydrothermal Vent
compounds. chemolithoautotroph, p. 93
Community (a) This
diverse community
A number of hydrothermal vents have been discovered, is supported by the
some thousands of meters below the ocean surface. These vents metabolic activities of
form when water seeps into cracks in the ocean chemolithoautotrophs.
floor and becomes heated by the molten rock, (b) Water escaping
from the vent is rich in
finally spewing out in the form of mineral-
reduced compounds,
laden undersea geysers. The hydrogen sulfide including hydrogen
discharged supports thriving deep-sea com- sulfide, which can serve
munities, oases in the otherwise desolate ocean as energy sources.
floor (figure 29.11). Large numbers of sulfur- ? Photosynthetic
oxidizing chemolithoautotrophs (bacteria and organisms are
archaea) are found in and around the vents. Many normally considered
are free-living but some live in symbiotic association with the the most important
primary producers;
large tube worms and clams that inhabit the areas. The chemo- which organisms are
lithoautotrophs obtain energy by oxidizing hydrogen sulfide, and primary producers in
they fix CO2, providing the animals with both a carbon and energy hydrothermal vent
source. communities?
Microbial populations have been found almost 3 km under
the ground and in iron-rich volcanic rocks from nearly a thousand
meters below the surface of the Columbia River (see Perspective (a)
4.1). These organisms gain energy from hydrogen (H2) produced
in the subsurface. It has been estimated that if (and it is a big “if” H 2S
H 2S H 2S
at this point) most similar rocks contain microbes, there could H 2S
be as much as 2 × 1014 tons of underground microorganisms—
H2S
equivalent to a layer 1.5 meters thick over the entire land surface
of the earth! H 2S
H 2S + O2 + CO 2

MicroAssessment 29.4 Terminal

H 2S Energy Carbon
electron
Elements are recycled as organisms incorporate them to produce source source
acceptor
biomass, oxidize reduced forms as energy sources, and reduce
oxidized forms as terminal electron acceptors. Carbon fixation uses
atmospheric CO2 to produce organic material; the CO2 is regenerated Bacteria
during respiration and some fermentations. Prokaryotes are essential H 2S and archaea
for several steps of the nitrogen cycle including nitrogen fixation,
nitrification, denitrification, and anammox. Prokaryotes are also SO 42- + Organic
essential for several steps of the sulfur cycle, including sulfur compounds
reduction and sulfate oxidation.
10. What are the three general roles of carbon-containing
compounds in metabolism?
11. Why do farmers try to prevent nitrification?
12. Although chemoautotrophs serve as the primary producers near
hydrothermal vents, animals there still ultimately depend on the
photosynthetic activities of plants and cyanobacteria. Why? +

29.5 ■ Mutualistic Relationships (b)

Between Microorganisms
and Eukaryotes
As described in chapter 16, mutualism is a symbiotic association
Learning Outcome
in which both partners benefit. A variety of other ecologically
11. Describe the mutualistic relationships between fungi and plant
important symbiotic relationships exist, but mutualistic relation-
roots, symbiotic nitrogen-fixers and plants, and microorganisms
and herbivores. ships highlight the vital role of microorganisms to life on this
planet. symbiosis, p. 381
730 Chapter 29 Microbial Ecology
Ectomycorrhizal
Root cells of plants fungus
Mycorrhizas (cross-section)

Mycorrhizas are fungi growing in symbiotic relationships with

plant roots (figure  29.12). They enhance the competitiveness of
plants by helping them take up phosphorus and other substances
from the soil. In turn, the fungi gain nutrients for their own growth
from root secretions. It is estimated that over 85% of vascular
plants (plants with specialized water and food conducting tissues)
have mycorrhizas.
There are two common types of mycorrhizal relationships:
■ Endomycorrhizas. The fungi penetrate root cells, growing
as coils or tight, bushlike masses within the cells. These are
by far the most common mycorrhizal relationships and are
found in association with most herbaceous plants. Relatively
few species of fungi are involved, perhaps only 100 or so, and
most appear to be obligate symbionts. The relationship for
some plants is also obligate; for example, most orchid seeds Endomycorrhizal fungus
will not germinate without the activities of a fungal partner. (a) Diagram of Mycorrhizas
■ Ectomycorrhizas. The fungi grow around the plant cells,
forming a sheath around the root. These fungi mainly associ-
ate with certain trees, including conifers, beeches, and oaks.
Over 5,000 species of fungi are involved in ectomycorrhizal
relationships but many are restricted to a single type of plant.

MicroByte
Chanterelles and truffles are examples of commercially valuable
ectomycorrhizal fungi.

Symbiotic Nitrogen-Fixers and Plants

Although some free-living bacteria can add fixed nitrogen to the
soil, symbiotic nitrogen-fixing organisms are far more significant
in benefiting plant growth and crop production. They are impor-
tant in both terrestrial and aquatic habitats.
(b) Endomycorrhiza

Rhizobia
Members of a diverse group of genera, including Rhizobium,
Bradyrhizobium, Sinorhizobium, and Azorhizobium—collectively
referred to as rhizobia—are the most agriculturally important
symbiotic nitrogen-fixing bacteria. These grow as endosymbi-
onts within specialized organs called nodules on the roots of
legumes (plants that bear seeds in pods) including alfalfa, clover,
peas, beans, and peanuts (figure  29.13a). The input of soil
nitrogen from rhizobia may be about 10 times the annual rate of
nitrogen fixation by non-symbiotic organisms. To foster plant
growth, farmers often add the appropriate symbionts to the seeds
of certain legumes. endosymbiont, p. 76
The plant cells foster the nitrogen-fixation of the endosymbi-
onts by synthesizing a protein called leghemoglobin. This binds
to O2 and regulates its concentration in the nodule, protecting the (c) Ectomycorrhiza
O2-sensitive nitrogenase. The plant also provides various nutrients
FIGURE 29.12 Mycorrhizas (a) Diagram illustrating the two
to the endosymbionts. Meanwhile, the endosymbionts provide types of mycorrhizas. (b) In an endomycorrhizal relationship,
fixed nitrogen to the plant. the fungi penetrate root cells growing within the cells. (c) In an
Nodule formation involves extensive chemical communica- ectomycorrhizal relationship, the fungi grow around the plant
tion between the rhizobia and legume partners (figure  29.13b). root cells, forming a fungal sheath around the root.
First, plant root secretions attract the appropriate rhizobial species, ? Which type of mycorrhizal relationship is common in herbaceous
which then colonizes the roots. In the most well-characterized plants?
 Part V Applied Microbiology 731

(a)

Chemical signals Bacteroids Mature

produced by within nodule
plant symbio-
Infection somes
thread
Nod factors
(NFs) produced Symbiosome
by rhizobia

Plant-bacterial communication Root hairs curl Bacteria pass through Nitrogen-fixing nodule
Chemicals produced by the plant NFs cause the root hairs to curl infection thread Rhizobia within the
cause rhizobia to colonize the around the bacterial cells and The root hair’s plasma membrane symbiosomes differentiate to
roots and produce Nod factors induce plant cell division. invaginates, forming an infection become nitrogen-fixing
(NFs). thread through which the rhizobia bacteroids.
pass. The dividing plant cells
engulf the rhizobia, enclosing
them within symbiosomes.
(b)
FIGURE 29.13 Root Nodules (a) Appearance. (b) The major steps leading to nodule formation by Rhizobium species.
? The pinkish color of the nodules is due to leghemoglobin. What is the role of this protein?

nodulation systems, some of the chemicals cause the bacteria to

produce Nod factors (NFs), which induce a series of events that Other Nitrogen-Fixing Symbionts
cause the root hair to curl, trapping the bacterial cells. The NFs Several genera of non-leguminous trees, including alder and
also cause underlying plant cells to divide. The plasma membrane gingko, have nitrogen-fixing root nodules at some stages of their
of the curled root hair then invaginates, forming a tubelike struc- life cycle. The bacteria involved in the symbiosis are members of
ture called an infection thread. The rhizobia cells travel through the genus Frankia.
this structure to underlying plant tissues, and are then engulfed In aquatic environments, the most significant nitrogen-fixers
by the dividing plant cells, forming membrane-bound structures are cyanobacteria. They are especially important in flooded soils
called symbiosomes. Within a symbiosome, a rhizobial cell such as rice paddies. In fact, rice has been cultivated successfully
changes in shape and function to become a specialized nitrogen- for centuries without the addition of nitrogen-containing fertilizer
fixing cell called a bacteroid. because of the symbiotic relationship between the cyanobacterium
Although the relationship between the plant and bacterium Anabaena azollae and the aquatic fern Azolla. The bacterium
is not obligate, it offers a competitive advantage to both partners. grows in specialized sacs in the leaves of the fern, providing
The rhizobia do not fix nitrogen in soils lacking legumes, and they nitrogen to the fern. Before planting rice, the farmer allows the
compete poorly with other microbes, slowly disappearing from flooded rice paddy to overgrow with Azolla ferns. Then, as the
soils in which legumes are not grown. Likewise, legumes compete rice grows, it eventually crowds out the ferns. As the ferns die and
poorly against other plants in heavily fertilized soils. decompose, their nitrogen is released into the water.
732 Chapter 29 Microbial Ecology

Microorganisms and Herbivores After being digested in the rumen, the food mass enters
another compartment (the omasum), eventually reaching the acidic
Another mutualistic relationship occurs between microbes and
true stomach (abomasum). There, more organic acids are absorbed.
certain herbivores. In order to subsist on grass and other plant
In addition, lysozyme is secreted, allowing the animal to lyse and
material, herbivores such as cattle and horses rely on a microbial
then digest members of the microbial population, providing even
community that inhabits a specialized digestive compartment. The
more nutrients. A critical feature of ruminants is that the microbial
microbes digest cellulose and hemicellulose, two of the major
population gets the first opportunity to use the ingested nutrients.
components of plant material, releasing compounds that can then
The animal then uses the metabolic end products as well as the
serve as a nutrient source for the animal. The specialized diges-
microbial cells themselves.
tive compartment in ruminants such as cattle, sheep, and deer is
The cecum of non-ruminant herbivores serves a function similar
called a rumen, and it precedes the true stomach. Non-ruminant
to a rumen. Microbes in the cecum cannot be used as a food source,
herbivores such as horses and rabbits have a compartment called
however, because of the cecum’s location relative to the stomach.
a cecum, which serves a similar purpose; it lies between the small
The benefit of the location is that the animal can digest and absorb
intestine and the large intestine. cellulose, p. 32
readily available nutrients without competition from microbes.
The rumen functions as an anaerobic fermentation vessel to
which nutrients in the form of plant materials are intermittently
added. In some cases, the animal produces over 150 liters of saliva MicroAssessment 29.5
per day. In addition to providing water, the saliva also contains
bicarbonate, a buffer, which helps maintain the pH. A remarkably Mycorrhizal fungi gain nutrients from plant root secretions while
helping plants take up substances from soil. Symbiotic nitrogen-fixers
complex variety of microorganisms degrade the ingested mate-
provide plants with a source of usable nitrogen while being provided
rial, releasing sugars that are then fermented, producing various with an exclusive habitat. Microorganisms in the rumen and cecum of
organic acids. Each milliliter of rumen content contains approxi- herbivores digest cellulose and hemicellulose, allowing the animal to
mately 1010 bacteria, 106 protozoa, and 103 fungi. Of the over 200 subsist on plant material.
species identified in the rumen, no single one accounts for more 13. Describe the differences between an endomycorrhiza and
than 3% of the total microbiota. The organic acids released during ectomycorrhiza.
fermentation are absorbed by the cells that line the rumen, provid- 14. Describe the differences between a rumen and a cecum.
ing the animal with an energy and nutrient source. Large quanti- 15. Gardeners sometimes plant clover between productive growing
ties of gas—produced as a result of fermentation—are discharged seasons. Why would this practice be beneficial? +
when the animal belches.

Summary
29.1 ■ Principles of Microbial Ecology Studying Microbial Ecology
Ecosystems vary in their biodiversity and biomass. The microenviron- Microbial ecology has been difficult to study because so few envi-
ment is most relevant to a microorganism’s survival and growth. ronmental prokaryotes can be successfully grown in the laboratory.
Molecular techniques, including fluorescence in situ hybridization
Nutrient Acquisition
(FISH), polymerase chain reaction (PCR), denaturing gradient gel elec-
Primary producers convert CO2 into organic material; consumers trophoresis (DGGE), and DNA sequencing are allowing researchers
use the organic materials produced by plants; decomposers digest the to better understand complex microbial communities. Soil and water
remains of primary producers and consumers (figure 29.1). metagenomes are being studied.
Microbes in Low-Nutrient Environments
29.2 ■ Aquatic Habitats
Microorganisms capable of growing in dilute aqueous solutions are
Oligotrophic waters are nutrient poor; eutrophic waters are nutrient
common in nature; often they grow in biofilms.
rich (figure 29.5). Excessive growth of aerobic heterotrophs may cause an
Microbial Competition and Antagonism aquatic environment to become hypoxic, resulting in the death of fish
Microorganisms in the environment vie for the same limited pool of and other aquatic animals.
nutrients (figure  29.2). A species can competitively exclude others, or Marine Environments
produce compounds that inhibit others.
Ocean waters are generally oligotrophic and aerobic, but inshore areas
Microorganisms and Environmental Changes can be dramatically affected by nutrient-rich runoff (figure 29.6).
Environmental changes are common, and often result in different spe- Freshwater Environments
cies becoming dominant (figure 29.3). Oligotrophic lakes may have anaerobic layers due to thermal
stratification. Shallow, turbulent streams are generally aerobic.
Microbial Communities
A microbial mat is a thick, dense, highly organized biofilm com- Specialized Aquatic Environments
posed of distinct colored layers of different groups of microbes Salt lakes and mineral-rich springs support the growth of microbes
(figure 29.4). specifically adapted to thrive in these specialized environments.
 Part V Applied Microbiology 733

29.3 ■ Terrestrial Habitats Phosphorus Cycle and Other Cycles

Characteristics of Soil Most plants and microorganisms take up orthophosphate, incorporating

it into biomass. Iron, calcium, zinc, manganese, cobalt, and mercury are
Soil represents an environment that can change abruptly and dramatically.
recycled by microorganisms.
Microorganisms in Soil Energy Sources for Ecosystems
The environmental conditions affect the density and composition of the Photosynthetic organisms convert radiant energy to chemical bond
soil microbiota. energy in the form of organic compounds. Chemolithoautotrophs har-
vest energy from reduced inorganic chemicals (figure 29.11).
The Rhizosphere
The concentration of microbes in the rhizosphere is generally much 29.5 ■ Mutualistic Relationships Between
higher than that of the surrounding soil. Microorganisms and Eukaryotes
29.4 ■ Biogeochemical Cycling and Energy Flow Mycorrhizas
Organisms use elements in biosynthesis to produce biomass, as sources Mycorrhizas help plants take up phosphorus and other substances from
of energy, and as terminal electron acceptors. soil; in turn the fungal partners gain nutrients for their own growth
(figure  29.12). Endomycorrhizal fungi penetrate root cells; ectomycor-
Carbon Cycle (figure 29.7)
rhizal fungi grow around root cells.
One of the fundamental aspects of the carbon cycle is carbon fixation.
As consumers and decomposers degrade organic material, respiration Symbiotic Nitrogen-Fixers and Plants
and some fermentations release CO2. Rhizobia reside as nitrogen-fixing endosymbionts in nodules on
legume roots (figure  29.13). Frankia species fix nitrogen in nodules of
Nitrogen Cycle (figure 29.9) alder and gingko. A species of cyanobacteria fixes nitrogen in special-
The steps of the nitrogen cycle include nitrogen fixation, ized sacs in the leaves of the Azolla fern.
ammonification, nitrification, denitrification, and anammox.
Microorganisms and Herbivores
Sulfur Cycle (figure 29.10) In order to subsist on grass and other plant material, herbivores rely on
Certain steps of the sulfur cycle—sulfur reduction and sulfate a community of microbes that inhabit a specialized digestive compart-
oxidation—depend on the activities of prokaryotes. ment, either a rumen or a cecum.

Review Questions
Short Answer 2. Which of the following is false?
1. Describe why a microbial mat has green, reddish-pink, and black a) Culture techniques are an accurate way of determining which
layers. members in a microbial community are most common.
2. Why do lakes in temperate regions stratify during the summer b) Fluorescence in situ hybridization (FISH) can be used to
months? distinguish subsets of prokaryotes that contain a specific
3. Why is there a high concentration of microbes in the rhizosphere? nucleotide sequence.
c) Polymerase chain reaction (PCR) can be used to distinguish
4. What dictates whether a form of an element is suitable for use as
subsets of prokaryotes based on their 16S rRNA sequences.
an energy source versus a terminal electron acceptor?
d) Denaturing gradient gel electrophoresis (DGGE) can be used to
5. Why does wood resting at the bottom of a bog resist decay?
separate PCR products.
6. What is the importance of nitrogen fixation? e) Studying the genome of one organism can give insights into the
7. Describe the relationship between ammonia oxidizers and nitrite characteristics of another.
oxidizers. 3. Which of the following pairs that relate to aquatic environments
8. How do hydrothermal vents support thriving communities of does not match?
microbes, clams, and tube worms? a) Oligotrophic—nutrient poor
9. Give examples of free-living and symbiotic nitrogen-fixing micro- b) Hypoxic—oxygen poor
organisms. Are these prokaryotic or eukaryotic? c) Epilimnion—O2 poor
10. Describe the steps that lead to the formation of the symbiotic rela- d) Hypolimnion—lower layer
tionship between rhizobia and legumes.
e) Eutrophic—nutrient rich.
4. Adding high levels of nutrients to a lake or inshore area would
Multiple Choice have all of the following effects in that environment except
1. Cyanobacteria are a) death of clams and crabs.
a) primary producers. b) consumers. b) increased growth of heterotrophic microbes.
c) herbivores. d) decomposers. c) increased growth of photosynthetic organisms.
e) more than one of the above. d) increased levels of dissolved O2.
734 Chapter 29 Microbial Ecology

5. Which of the following pairs that relate to terrestrial environments soil bacteria. The ad claimed that soil bacteria were responsible
does not match? for most crop losses. The farmer called the agricultural extension
a) Soil—minimal biodiversity office at a local university for advice. Explain what the extension
b) Bacillus—endospores office adviser most likely told the farmer about the usefulness of
c) Streptomyces—geosmin production the product.
d) Fungi—lignin degradation 2. Recent reports suggest that human activities, such as the generous
e) Rhizosphere—soil that adheres to plant root use of nitrogen fertilizers, have doubled the rate at which elemental
nitrogen is fixed, raising concerns of environmental overload of
6. Atmospheric nitrogen can be used
nitrogen. What problems could arise from too much fixed nitrogen,
a) directly by all living organisms. and what could be done about this situation?
b) only by aerobic bacteria.
c) only by anaerobic bacteria. Critical Thinking +
d) in symbiotic relationships between rhizobia and plants. 1. Each colony growing on an agar plate arises from a single cell (see
e) in photosynthesis. photo). Colonies growing close together are much smaller than
7. Which process converts ammonium (NH4+) into nitrate (NO3–)? those that are well separated. Why would this be so?
a) Nitrogen fixation b) Ammonification c) Nitrification
d) Denitrification e) Anammox
8. Energy for ecosystems can come from
a) sunlight via photosynthesis.
b) oxidation of reduced inorganic chemicals by chemoautotrophs.
c) both a and b.
9. Mycorrhizas represent associations between plant roots and micro-
organisms that
a) are antagonistic.
b) help plants take up phosphorus and other nutrients from soil.
c) involve algae in the association with plant roots.
d) form nodules on the plant’s leaves.
e) lead to the production of antibiotics.
10. In symbiotic nitrogen fixation by rhizobia and legumes 2. An entrepreneur found an economically feasible way of collect-
a) the amount of nitrogen fixed is much greater than by non- ing large amounts of sulfur from underwater hot vents in the
symbiotic organisms. Pacific Ocean. The sulfur will be harvested from the microorgan-
b) neither the bacteria nor the legume can exist independently. isms found in the vent areas. A group of ecologists argued that
c) the bacteria enter the leaves of the legume. the project would destroy the fragile ecosystem by depleting it
of usable sulfur. The entrepreneur argued that the environment
d) the bacteria operate independently of the legume.
would not be harmed because the vents produce more than
Applications enough sulfur for the clams and tube worms in the area. Explain
who is correct.
1. A farmer who was growing soybeans, a type of legume, saw an
Internet site advertising an agricultural product for safely killing
30 Environmental Microbiology:
Treatment of Water, Wastes, and Polluted Habitats

KKEEYYTTEERRMMS S
Advanced Treatment Any
physical, chemical, or biological
purification process beyond
secondary treatment of wastewater.
Sanitary Landfill A site used for
disposal of non-hazardous solid
wastes in a manner that minimizes
damage to human health and the
environment.
Anaerobic Digestion Process
that uses anaerobic microbes to Secondary Treatment
degrade the sludge obtained during A biological process in which
wastewater treatment. microbial growth is actively
encouraged in wastewater, allowing
Biochemical Oxygen Demand
the microbes to convert suspended
(BOD) The amount of O2 required
solids to inorganic compounds and
for the microbial decomposition of
removable cell mass.
organic matter in a sample.
Septic System An individual
Bioremediation Process that uses
wastewater treatment system in
microorganisms to degrade harmful
which the sludge settles out in a
chemicals.
large tank where it is degraded by
Composting The natural microorganisms and the effluent
decomposition of organic solid percolates through a drain field.
material.
Sludge The solid portion of
Effluent The liquid portion of wastewater that settles to the
treated wastewater. bottom of sedimentation tanks
Indicator Organisms Microbes during primary and secondary
commonly found in the intestinal treatment.
A pristine mountain lake.
tract whose presence in other Wastewater Material that flows
environments suggests fecal from household plumbing systems;
contamination. municipal wastewater also includes
A Glimpse of History Primary Treatment A physical business and industrial wastes and
storm water runoff.
process designed to remove materials
Delivering fresh water to urban areas and removing human wastes have that will settle out of wastewater.
been practiced at least since Roman times. The ruins of aqueducts that
delivered fresh water long distances can be seen today in many parts of
Europe. Removing human wastes from cities has been more difficult,
however, and the sewers that were used until the mid-nineteenth century
were not much more than large, open cesspools.
Long before the discovery of the microbial world, people recog- the lines envisioned by Chadwick. New York City did the same in
nized that some diseases are associated with water supplies. As early as 1866, again in response to a threatened cholera epidemic. By the end of
330 b.c., Alexander the Great had his armies boil their drinking water, the nineteenth century, most large European and U.S. cities had sewer
a habit that probably contributed to his huge successes. Certainly, many systems to remove and treat waste materials as well as water systems
battles have been lost over the years as a result of waterborne diseases to deliver safe drinking water. Cholera in the industrialized nations of
that sickened or killed the soldiers. Years before Vibrio cholerae was Europe and North America virtually disappeared.
identified as the cause of cholera, people recognized that cholera epidem-
ics were associated with drinking water. The desire for clean, clear water

M
 
led to sand filtration systems being used in London and elsewhere in the ost people living in developed countries take for granted
early nineteenth century. Late in that century, Robert Koch showed that that their tap water is safe to drink, their wastes will
this kind of filtration yielded clear water and also removed nearly all reliably disappear into sewers or landfills for proper
bacteria from the water. disposal, and pollutants (substances that are harmful or injurious)
As early as the 1840s, Edwin Chadwick, an English activist, cham- will not accumulate in the environment. They seldom consider the
pioned a new idea on how wastes could be removed. His idea was to
role that microbes play in these essential aspects of modern life.
construct a system of narrow, smooth ceramic pipes through which water
Microorganisms are important in the treatment processes
under pressure could be flushed along with solid waste materials. This
system would carry the wastes away from the inhabited part of the city described in this chapter for two very distinct reasons. First, we
to a distant collection site. There, he hoped to collect them and produce benefit from the fact that microbes are the ultimate recyclers,
fertilizer to sell to farmers. playing an essential role in the decomposition of our wastes. At
In 1848, concerned about cholera, the Board of Health in England the same time, pathogens must be eliminated from sewage before
started widespread reforms and began installing a sewage system along it is discharged and from drinking water for it to be potable (safe

735
736 Chapter 30 Environmental Microbiology

for human consumption). Recreational waters such as swimming indicate that large amounts of degradable materials are present,
pools, water parks, lakes, rivers, and shorelines are also monitored resulting in correspondingly large amounts of O2 being consumed
to ensure they do not have harmful levels of certain pathogens. during its biological degradation. The BOD of raw sewage is
Treatment of water, waste, and polluted habitats is a signifi- approximately 300 to 400 mg/liter, which could easily deplete the
cant challenge, particularly in densely populated areas. Consider dissolved O2 in the receiving water. The dissolved O2 content of
that every day the average American uses about 150 gallons of natural waters is generally 5 to 10 mg/liter.
water, and generates 120 gallons of wastewater and 5 pounds To determine the BOD, the O2 level in a well-aerated sample
of trash. This means that a city with only 1 million inhabitants of microbe-containing test water is first measured. The sample is
is faced with the disposal of approximately 44 billion gallons of then incubated in a sealed container in the dark under standard
wastewater and a million tons of trash each year! conditions of time and temperature, usually 5 days at 20°C. The
O2 level is then determined again. The difference between the dis-
solved O2 at the beginning of the test and at the end reflects the
30.1 ■ Microbiology of BOD of the sample. In many cases, the sample must be diluted
first in order to accurately determine the BOD.
Wastewater Treatment
Learning Outcomes Municipal Wastewater
1. Describe the concept of BOD. Treatment Methods
2. Compare and contrast primary treatment, secondary treatment, Large-scale wastewater treatment plants in the United States
advanced treatment, and anaerobic digestion. use a series of two processes—primary and secondary
3. Describe how septic systems function. treatment—as required by the 1972 Federal Water Pollution
Control Act, now known as the Clean Water Act. Once treated,
Wastewater, or sewage, is composed of all the material that flows the effluent (the liquid portion) can be discharged into the
from household plumbing systems, including washing and bathing receiving water. Additional steps are used to treat sludge
water and toilet wastes. Municipal wastewater also includes busi- (the solid portion).
ness and industrial wastes. In many cities, storm water runoff that
flows into street drains enters the system as well. Primary Treatment
The most obvious reason that wastewater must be treated Primary treatment is a physical process designed to remove
before discharge is that pathogenic microbes—including those that materials that will settle out, removing approximately 50% of the
cause diarrheal diseases and hepatitis—can be transmitted in feces. solids and 25% of the BOD. Raw sewage is first passed through
If untreated sewage is released into a river or lake that is then used a series of screens to remove large objects such as sticks, rags,
as a source of drinking water, disease can easily spread. If marine and trash (figure  30.1). Skimmers then remove scum and other
waters become contaminated in a similar manner, eating the local floating materials. The sewage then sits in a sedimentation tank,
shellfish can result in disease. Shellfish are filter feeders and they allowing solids to settle out. Once the settling period is complete,
concentrate microbes from the waters in which they live. the sludge is removed and the primary treated wastewater that
The high nutrient content of wastewater can also be damag- remains is sent for secondary treatment.
ing to the receiving water. When any nutrient-rich substance is
added to an aqueous environment, microorganisms quickly use the Secondary Treatment
compounds as energy sources, breaking them down in metabolic
Secondary treatment is chiefly a biological process that converts
pathways such as glycolysis and the TCA cycle (see figure 6.10).
most of the suspended solids to inorganic compounds and cell
As a result, microbes that aerobically respire consume available
mass that can then be removed, eliminating as much as 95% of the
O2 in the water, using it as a terminal electron acceptor (see figure
BOD. Microbial growth is actively encouraged during secondary
6.20). The amount of dissolved O2 in lakes and rivers is limited
treatment, allowing aerobic organisms to oxidize the biologically
and can easily be depleted during the microbial breakdown of
degradable organic material to CO2 and H2O. Because secondary
nutrients. Fish and other aquatic animals in the environment die
treatment relies on the metabolic activities of microorganisms, the
because they require O2 for respiration (see figure 29.6). Thus,
processes could be devastated if too much toxic industrial waste
effective wastewater treatment must decrease the level of organic
or hazardous household materials were dumped into wastewater
compounds substantially, in addition to eliminating pathogens and
systems, killing the microbial population.
pollutants. aerobic respiration, p. 134
Methods used for secondary treatment include:
■ Activated sludge process. This common system relies on
Biochemical Oxygen Demand (BOD) mixed populations of aerobic microbes that grow as flocs
An important goal of wastewater treatment is to decrease the envi- (suspended biofilms). Although the organisms are often
ronmental impact by reducing the biochemical oxygen demand naturally present in wastewater, large numbers are added
(BOD)—the amount of O2 required for the microbial decom- by introducing a small portion of leftover sludge from the
position of organic matter in a given sample. High BOD values previous load of treated wastes. Plenty of O2 is supplied
 Part V Applied Microbiology 737

Primary Treatment—
physical removal of organic material
Anaerobic Sludge Digestion—
anaerobic microbial degradation of organic material
Sedimentation tank
Screens
Raw
sewage

Effluent

Sludge

Drying bed
Anaerobic digester

Disposal, incineration, or
use as a soil enhancer
Secondary Treatment—
microbial degradation of organic material

Activated sludge process

Option 1
Advanced Treatment—
additional purifying treatments
Air or O2
Aeration tank (chemical, physical, or microbial)
Sedimentation tank
Option A
To sludge Treatments vary, but may include precipitation
digestion of phosphates, removal of ammonia, and
Trickling filter system biological degradation of nitrates.

Option 2

Trickling filter Sedimentation tank

To sludge
digestion Option B Disinfection—
Lagoons destruction of pathogens and other microbes

Option 3 Treatments such as chlorination, UV light, ozone

Constructed wetlands Discharge to

receiving water

Option 4

FIGURE 30.1 Municipal Wastewater Treatment The processes consist of primary treatment, secondary treatment, advanced treatment
(optional), disinfection, and anaerobic sludge digestion.
? What is the purpose of secondary treatment?
738 Chapter 30 Environmental Microbiology

by mixing the wastewater in an aerator. As

2 Rotating arm
the microbes multiply, the organic matter is Distributes effluent
converted into both biomass and waste prod-
ucts such as CO2. Following the aeration, the
wastewater is again sent to a sedimentation
tank. There, most of the flocs settle and the
3 Filter media
resulting sludge is removed; a portion of this Biofilm-coated plastic pieces
sludge is introduced to a new load of waste- or coarse gravel and rocks
water to act as an inoculum. A complication
of the activated sludge process occurs when 4 Effluent trickles
through.
filamentous bacteria such as Thiothrix species
overgrow in the wastewater during treatment,
creating a buoyant mass that does not settle.
This problem, called bulking, interferes with
the separation of the solid sludge from the 5 Outlet pipe
liquid effluent. biofilm, p. 84 Thiothrix, p. 262 Effluent from the filter goes to
1 Inlet pipe
a sedimentation tank before
■ Trickling filter (TF) system. This method is Effluent from primary advanced treatment or disinfection.
frequently used in smaller wastewater treat- treatment enters
ment plants. The TF has a rotating arm that
distributes the effluent over a bed of plastic FIGURE 30.2 Trickling Filter Wastewater is channeled into the rotating arm,
and then trickles through holes in the bottom of the arm onto plastic or a gravel
pieces or coarse gravel and rocks (figure 30.2). and rock bed.
The surfaces of these materials become coated
with a biofilm—a mix of bacteria, fungi, ? What role do biofilms play in trickling filters?
algae, protozoa, and nematodes—that aerobi-
cally degrades the organic material as it passes. The rate of Advanced Treatment
wastewater flow can be adjusted for maximum degradation.
As water supplies are becoming scarce—and in order to comply
■ Lagoons. The wastewater is channeled into shallow ponds, or with discharge standards designed to protect the environment—
lagoons, where it remains for several days to a month or more, many communities are finding advanced treatment necessary.
depending on the design of the lagoon. Algae and cyanobac- This includes any purification process beyond secondary treatment;
teria that grow at the surface provide O2, allowing aerobic it may involve physical, chemical, or biological processes, or any
organisms in the ponds to degrade the organic materials. combination of these. Advanced treatment is expensive, however,
■ Constructed wetlands. These follow the same principles as and has not been common in the past.
lagoons, but their more advanced designs make them suitable Advanced treatment is often designed to remove ammo-
habitats for birds and other wildlife (figure 30.3). For exam- nia, nitrates, and phosphates—compounds that foster growth
ple, the wastewater treatment processes in Arcata, California, of algae and cyanobacteria in receiving waters. The concen-
use a series of marshes trations of these nutrients, which are very low in unpolluted
that now attract a variety waters, normally limit the growth of the photosynthetic organ-
of shorebirds and serve isms. Consequently, if the nutrients are added, photosynthetic
as a wildlife sanctuary. microbes proliferate, often leading to buoyant masses of cells

Aquatic plants FIGURE 30.3 Constructed

Wetland (a) Photograph.
Effluent (b) General components.
inlet
? What advantage does a
Effluent constructed wetland have
outlet over a lagoon?

Gradual slope to outlet

Soil or gravel Watertight membrane

(a) (b)
 Part V Applied Microbiology 739

PERSPECTIVE 30.1
What a Gas!
Inlet for wastes
In rural areas of China, fuel
for cooking and heating is
Gas delivery
often in short supply; as a
consequence, the hay that
should go for animal feed
must be used for fuel. To Fermentation tank
help solve this problem,
many of the farmers build
methane-producing tanks on
Overflow
their farms (figure 1). Near the tank
family house is an underground
cement tank connected to the Gas
latrine and pigpen. Human and
animal wastes along with water
and some other organic materi-
als such as straw are added to the Slurry
tanks. As the natural process of
decomposition occurs, methane gas
(CH4) is produced. This gas rises to
the top of the tank and is connected to the
house with a hose. Enough gas is produced
to provide lights and cooking fuel for the farm
family. By producing its own gas, a family also saves
money because it does not need to buy coal for cooking.
The effluent slurry that accumulates as the wastes decompose can
be removed and used as fertilizer. FIGURE 1 Methane-Producing Tank

that create a surface scum (see figure  11.7). Accumulations of Anaerobic Digestion
photosynthetic organisms also provide a source of carbon for Sludge removed during the sedimentation steps of primary and sec-
other microbes, increasing the BOD and, consequently, threaten- ondary treatment is transferred to a tank for anaerobic digestion.
ing other forms of aquatic life. There, various anaerobic microbes act sequentially, ultimately
Ammonia can be removed by a process called ammonia strip- converting much of the organic material to methane:
ping, which liberates gaseous ammonia from the water. Nitrates
can be removed using denitrifying bacteria. These organisms use ■ Organic compounds → organic acids, CO2, H2
nitrate as a terminal electron acceptor during anaerobic respira- ■ Organic acids → acetate, CO2, H2
tion, forming nitrogen gas. This gas is inert, non-toxic, and easily
■ Acetate, CO2, H2 → methane (CH4)
removed. The discovery of anammox bacteria, which use ammo-
nia as an energy source and nitrite as a terminal electron acceptor, Many wastewater treatment plants are equipped to use the meth-
provides an alternative means of removing inorganic nitrogen- ane generated, thereby avoiding the cost of other sources of
containing compounds. Phosphates are eliminated using chemi- energy to run their equipment (see Perspective 30.1).
cals that combine with phosphates, causing them to precipitate. After anaerobic digestion, water is removed from the remain-
denitrification, p. 727 anammox, p. 727 ing sludge, generating a nutrient-rich product called stabilized
sludge. This can be incinerated or disposed of in landfills but
Disinfection may also be used to improve soils and promote plant growth. The
Before discharge into the receiving water, the effluent is disin- sludge generated by the city of Milwaukee, Wisconsin, is used to
fected with chlorine, ozone, or UV light to decrease the numbers produce Milorganite, a fertilizer for lawns, gardens, golf courses,
of microorganisms and viruses. If chlorine is used, the disinfected and playfields. An increasing number of wastewater treatment
water can then be dechlorinated to avoid releasing excessive facilities are finding similar ways to recycle their treated sludge.
amounts of the toxic chemical into the environment. chlorine, Concerns exist, however, about heavy metals and other pollutants
p. 118 ozone, p. 120 UV light, p. 115 that can sometimes be concentrated in the product.
740 Chapter 30 Environmental Microbiology

MicroAssessment 30.1
Individual Wastewater
Primary treatment is a physical process that removes material that will
Treatment Systems settle out. Secondary treatment is chiefly a biological process that converts
Rural dwellings customarily rely on septic systems for waste- suspended material into inorganic compounds and microbial biomass.
water treatment (figure  30.4). Household wastewater is first Advanced treatment is often designed to remove ammonia, phosphates,
directed to a large tank, where much of the solid material settles and nitrates. Anaerobic digestion converts much of the organic matter to
methane. Septic systems are individual wastewater treatment systems.
and is degraded by anaerobic microorganisms. The effluent—
which has a high BOD—then flows to a series of perforated pipes, 1. What does the term BOD mean? What is its significance?
where it percolates (slowly passes) through a gravel-containing 2. What is the advantage of removing phosphates and nitrates
drain field. The drain field is designed to allow aerobic micro- from wastewater?
organisms to oxidize the organic material in the same manner 3. Why is denitrification unlikely to occur during
described for the trickling filter. secondary treatment? +
Septic systems must be properly designed and monitored
to ensure they work adequately. Circumstances that prevent
adequate drainage—such as a clay soil under a drain field— 30.2 ■ Drinking Water Treatment
allow anaerobic conditions to develop, preventing the organic and Testing
material from being oxidized adequately. In addition, toxic
materials can inhibit microbial activity in the system. Drainage Learning Outcomes
from a septic tank may contain pathogens; therefore, the tank 4. Describe how drinking water is typically treated.
must never be allowed to drain where it can contaminate water 5. Describe why and how drinking water is tested for total coliforms.
supplies.

FIGURE 30.4 Septic System (a) The components of a septic Inspection access Access for cleaning
system. (b) Septic tank, where anaerobic degradation takes place. Inspection access
? Why is it important that the drainage fields are aerobic?
Wastewater
from house

Scum
Outlet to
drain field

Effluent

Sludge

(b)

Household
wastewater Distribution box
Drain field
Septic tank

Vent pipe
Perforated pipe
Gravel

(a)
 Part V Applied Microbiology 741

Large cities generally obtain their drinking water from surface

waters such as lakes or rivers. Because surface water may serve Water Treatment Processes
as the receiving water for another city’s wastewater effluent, Community drinking water treatment is designed to eliminate
drinking water treatment is intimately connected to wastewater pathogenic microbes as well as harmful chemicals. First, water
treatment. The quality of the surface water is also affected by flows into a reservoir and is allowed to stand long enough for the
the characteristics of the watershed (the land over which water particulate matter to settle out (figure  30.6). The water is then
flows into the river or lake). Even pristine rivers are likely con- transferred to a tank where it is mixed with a chemical that causes
taminated with feces of animals that inhabit the watershed. suspended materials to coagulate (flocculate); an example of a
Smaller communities often use groundwater—pumped from coagulant is alum (aluminum sulfate). The mixture then flows to a
a well—as a source of drinking water. Groundwater is in aqui- sedimentation tank, where the coagulated materials are allowed to
fers (water-containing layers of rock, sand, and gravel) and is slowly sink to the bottom. As they settle, some microbes and other
replenished as water from rain and other sources seeps through substances are trapped and thereby removed as well.
the soil. Because aquifers are not directly exposed to animals and Following the removal of the coagulated materials, the water
the atmosphere, they are somewhat protected from contamination. is filtered—often through a thick bed of sand and gravel—to
However, poorly located or maintained septic systems and sewer remove various microbes including bacteria and protozoan cysts
lines, as well as sludge or other fertilizers, can lead to groundwater and oocysts. Organic chemicals that may be harmful or give
contamination (figure 30.5). undesirable tastes and odors can be removed by additional filtra-
Public water systems in the United States are regulated under tion. This is done using an activated charcoal filter, which adsorbs
the Safe Drinking Water Act of 1974, amended in 1986 and dissolved chemicals. An added benefit of filtration is that micro-
1996. This gives the Environmental Protection Agency (EPA) the organisms growing in biofilms on the filter materials use carbon
authority to set standards in order to control the level of contami- from the water as it passes. This lowers the organic carbon content
nants in drinking water. Standards are modified in response to new of the water, resulting in less microbial growth in pipes delivering
concerns; for example, regulations now govern the maximum lev- the water. filtration, p. 114
els of Cryptosporidium oocysts, Giardia cysts, and enteric viruses Finally, the water is treated with chlorine or other disinfec-
in drinking water. Cryptosporidium, p. 604 Giardia, p. 602 tants to kill or inactivate harmful bacteria, protozoa, and viruses

Rainfall

Wastes Irrigation
Drinking
water
well
Sludge
Fertilizer
Septic tank
Soil

Sewer line

Defective
well
lining

Groundwater

FIGURE 30.5 Groundwater Contamination Poorly located or maintained septic tanks and sewer lines, as well as sludge or other fertilizers,
can lead to groundwater contamination.
? How is water in the aquifer replenished?
742 Chapter 30 Environmental Microbiology

coliforms—lactose-fermenting members of the fam-

1 Settling ily Enterobacteriaceae, including Escherichia coli.
Large materials settle out.
The group is functionally defined as facultatively
Raw water reservoir anaerobic, Gram-negative, rod-shaped, non-spore-
forming bacteria that ferment lactose, forming acid
Coagulant added and gas within 48 hours at 35°C.
2 Coagulation
Although total coliforms are routinely present
Alum and other added chemicals in the intestinal contents of warm-blooded animals,
Flocculation tank combine with suspended material certain species can also thrive in soils and on plant
to form clumps.
material. Because of this, their presence does not
necessarily imply fecal pollution. To compensate for
3 Sedimentation this shortcoming, fecal coliforms, a subset of total
The coagulated materials coliforms more likely to be of intestinal origin, are
settle to the bottom.
also used as indicator organisms. The most common
Sedimentation tank fecal coliform is E. coli. Note that although some
E. coli strains can cause intestinal disease, the spe-
cies is used in water testing merely to indicate fecal
4 Filtration pollution.
Sand and gravel filters remove microorganisms Methods used to detect total coliforms in a water
including protozoan cysts and oocysts. Activated
Filtration unit
charcoal filters removed dissolved chemicals.
sample include:
Disinfectant ■ ONPG/MUG test. A water sample is added
added
to a medium containing ONPG (o-nitrophenyl-
5 Disinfection and Storage
Chlorine, ozone, or other chemical β-D-galactopyranoside) and MUG (4-methyl-
disinfectants are added. UV irradiation umbelliferyl-β-D-glucuronide). Lactose-fermenting
may also be used to destroy microbes. bacteria hydrolyze ONPG, generating a yellow
Reservoir compound; thus, all coliforms turn the medium yel-
low (figure 30.7). E. coli produces an enzyme that
hydrolyzes MUG, making a fluorescent compound.
Because of ONPG and MUG, a sample can be
tested simultaneously for both total coliforms and
Consumer use
E. coli.
FIGURE 30.6 Steps in the Treatment of Municipal Water Supplies ■ Presence/absence test. A 100-ml water sample
? What methods are used to disinfect municipal water supplies? is added to a lactose-containing broth that con-
tains a vial to trap gas. If gas is produced, the
broth is then tested to confirm that coliforms are
that might remain. A concern with using chlorine, however, is present.
that some of the disinfection by-products might be carcinogenic. ■ Most probable number (MPN) method. This statistical
In response to this concern, ultraviolet irradiation and ozone are assay of cell numbers uses successive dilutions to deter-
increasingly being used as alternatives, but a small amount of mine the most probable number of bacteria in a sample (see
chlorine must still be added to prevent problems associated with figure 4.20). To test drinking water, the broth used is similar
post-treatment contamination. Note that disinfection of waters to that in the presence/absence test. Positive tubes are further
with a high organic content requires more chlorine because organic tested to confirm that they contain coliforms.
compounds consume free chlorine. chlorine, p. 118 disinfection ■ Membrane filtration. A water sample is passed through a
by-products, p. 110 ultraviolet irradiation, p. 116 ozone, p. 120 filter that retains bacteria (see figure 4.19), concentrating the
bacteria from a known volume of water. The filter is then
placed on a lactose-containing selective and differential agar
Water Testing medium. selective media, p. 95 differential media, p. 95
A primary concern regarding the safety of drinking water is the
possibility that it might be contaminated with any of a wide The total coliform rule establishes a maximum number of
variety of intestinal pathogens, such as those discussed in chapter positive samples (100 ml) permitted in water samples. That limit
24. It is not feasible to test for all of the pathogens, however, so depends on how many samples are routinely collected for moni-
indicator organisms function as surrogates. These microbes are toring, which relates to the size of the population served by the
routinely found in feces, survive longer than intestinal pathogens, water system. When at least 40 samples per month are collected,
and are relatively easy to detect. The most common group of the system is in violation if more than 5% test positive for total
bacteria used as indicator organisms in the United States is total coliforms in a month. If fewer samples are collected, the system
 Part V Applied Microbiology 743

In addition to ridding our environment of wastes in water, we

must dispose of the solid wastes (garbage) generated each day.
Eliminating these has become an increasingly complex problem.

Sanitary Landfills for Solid

Waste Disposal
Sanitary landfills are widely used to dispose of non-hazardous
solid wastes in a manner that minimizes damage to human health
and the environment. Before sanitary landfills were developed,
solid wastes were often piled up on the ground in open-burning
dumps, attracting insects and rodents and causing aesthetic and
public health problems.
Federal standards dictate that sanitary landfills must be
located away from wetlands, earthquake-prone faults, flood
plains, or other sensitive areas. The excavated site is lined with
FIGURE 30.7 ONPG/MUG Test Coliforms hydrolyze ONPG,
plastic sheets or a special membrane on top of a thick layer of clay
yielding a yellow-colored compound. E. coli hydrolyzes MUG, to prevent contaminates from seeping out into the surrounding
generating a blue fluorescent compound. environment. A layer of sand with drainage pipes is placed on top
? What is the significance of finding coliforms in a drinking of this. When wastes are added to the site, they are compacted and
water supply? covered with a layer of soil every day. Once a landfill is full, it
is covered with soil and plants and can be used for recreation and
eventually as a site for construction. Methane and other gases are
vented, and the methane is burned or recovered for use.
is in violation if more than one sample tests positive per month. Sanitary landfills have several disadvantages with respect
If a sample tests positive, repeat samples within 24 hours are to waste management. For one thing, only a limited number
mandated. Samples positive for total coliforms are also tested of sites are available for use near urban and suburban areas. In
for either fecal coliforms or E. coli. If a water system exceeds addition, the methane gas must be removed as the organic waste
the monthly total coliform limit, the state and the public must be material anaerobically decomposes, a process that can take more
notified. Notification is also required if either of two sequential than 50 years. If buildings are constructed before the methane is
samples that test positive for total coliforms also test positive for removed, disastrous gas explosions can occur. Pollutants such as
fecal coliforms or E. coli. heavy metals and pesticides can leak from landfill sites into the
Because total coliform and fecal coliform assays cannot underground aquifers. It is very difficult to purify these aquifers
always predict contamination with protozoan cysts and oocysts, once they have become contaminated.
alternatives are being explored. Other microbes that can be Sanitary landfills have traditionally been a low-cost method
used as indicators of fecal pollution include enterococci, some of handling large quantities of solid waste. Because of increased
Clostridium species, and certain types of bacteriophages. costs and decreased availability of land, however, many cities are
looking for ways to decrease the amount of solid waste dumped
MicroAssessment 30.2 in landfills. In some cities, the fees charged to people for gar-
bage collection are based on the size of the container collected.
Drinking water may be obtained from surface water or groundwater.
Treatment of drinking water is designed to eliminate pathogens and The smaller the can, the lower the cost. This is intended to raise
harmful chemicals. In the United States, total and fecal coliforms are people’s awareness of how much solid waste they are generating
the most commonly used indicator organisms. as well as offer an incentive to recycle. Programs to recycle paper,
4. What is the purpose of coagulation in drinking water treatment? plastics, glass, and metal are being implemented in many cities
and counties with great success. Through these programs, landfill
5. Describe two methods of water testing.
areas will be available for a longer period of time.
6. Which would be more likely to cause illness and why—a water
sample that tested positive for fecal coliforms or one that tested
positive for E. coli O157:H7? +
Municipal and Backyard Composting—
Alternative to Landfills
30.3 ■ Microbiology of Solid Composting is the natural decomposition of organic solid mate-
Waste Treatment rial. Municipal and home composting programs are becoming
popular in many areas and are succeeding in reducing the amount
Learning Outcome of organic wastes added to landfills. As an added benefit, the black
6. Compare and contrast sanitary landfills and composting programs. organic material generated is commercially valuable and can be
used to improve garden soils.
744 Chapter 30 Environmental Microbiology

Backyard composting usually starts with a supply of organic

material such as leaves, grass clippings, and kitchen wastes
(figure 30.8). Composting of meats and fats is generally not rec-
ommended because they attract rodents and other pests. Often,
some soil and water are added to facilitate the process. As a
result of microbial metabolism, the inside of the pile heats up.
At 55°C to 66°C, pathogens are killed but thermophilic organ-
isms are not affected. If the pile is frequently aerated, which
can be done by physically stirring and turning it, and it is kept
moist, the composting can be completed in as little as 6 weeks.
thermophile, p. 90
Composting on a large scale offers cities a way to reduce
the amount of garbage sent to their landfills. In some cities, yard
wastes are collected separately from the main garbage. These are
then processed using machinery such as grinders that help make
the composting more efficient (figure 30.9). FIGURE 30.9 Municipal Composting
? What is the purpose of grinding the yard waste before putting it
in piles?

MicroAssessment 30.3
Sanitary landfills are used to dispose of non-hazardous solid wastes.
Composting, the natural decomposition of organic solid material,
dramatically reduces the need for large landfills.
7. What are the advantages and disadvantages of landfills?
8. List the steps in successful composting.
9. Why would soil and water be added to a compost pile? +

30.4 ■ Microbiology
of Bioremediation
Learning Outcomes
7. Describe why pollutants are a problem, and why some persist in
(a)
the environment.
8. Describe how two bioremediation strategies remove pollutants.

Bioremediation is the use of microorganisms to degrade or detox-

ify pollutants in a given environment. It generally takes advantage
of organisms already present in the polluted environment, but in
some cases specific organisms are added to the environment.

Pollutants
Most naturally occurring organic compounds are biodegradable,
meaning they can be degraded by one or more species of micro-
organisms. As oil spills dramatically demonstrate, however, some
natural materials can cause devastating effects before they are
degraded.
Synthetic compounds are more likely to be biodegradable if
their chemical composition is similar to that of naturally occur-
(b) ring substances. In contrast, xenobiotics (synthetic compounds
FIGURE 30.8 Backyard Composting (a) A home compost bin. quite different from any in nature) are more likely to persist in the
(b) Garden debris and many kitchen organic wastes can be environment because microorganisms are unlikely to have suit-
composted. able enzymes to break them down. Such enzymes would not give
? Why is it important that the compost heap be turned frequently? a microbe a competitive advantage in a natural situation.
 Part V Applied Microbiology 745

Means of Bioremediation
Many factors influence the degradation rate of pollutants. As a
Relative herbicide concentration

general rule, any practice that favors multiplication of microor-

100 ganisms will increase the rate of degradation. Thus, providing
2,4,5-T adequate nutrients, maintaining the pH near neutrality, raising the
75 H O H O temperature, and providing an optimal amount of moisture are all
H C C OH H C C OH likely to promote pollutant degradation. environmental factors that
O O influence microbial growth, p. 89 nutritional factors that influence microbial
50 1 1
2 2 growth, p. 92
6 Cl 6 Cl
5 5 There are two general bioremediation strategies—
25 3 3
2,4-D 4 Cl 4 biostimulation and bioaugmentation. Biostimulation enhances
Cl Cl growth of local microbes in a contaminated site by providing addi-
0 tional nutrients. Petroleum-degrading bacteria are naturally present
5 10 20 25
in seawater, but they degrade oil at a very slow rate because the
Time (days)
low levels of certain nutrients, including nitrogen and phosphorus,
FIGURE 30.10 Comparison of the Rates of Disappearance of
limit their growth. To enhance bioremediation of oil spills, a fertil-
Two Structurally Related Herbicides, 2,4-D and 2,4,5-T izer containing these nutrients—and which adheres to oil—was
developed. When this fertilizer is applied to an oil spill, microbial
? Why might the additional chlorine atom in 2,4,5-T molecule result
in slower degradation of the molecule? growth is stimulated, leading to at least a threefold increase in the
speed of degradation (figure  30.11). Bioaugmentation relies on
Relatively slight molecular changes significantly alter the activities of microorganisms added to the contaminated material,
biodegradability of a compound. Perhaps the best-studied example complementing the resident population. The activated sludge pro-
involves the herbicides 2,4-dichlorophenoxyacetic acid (2,4-D) cess used during secondary treatment of wastewater is a form of
and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T). The only dif- bioaugmentation. A great deal of research is underway to develop
ference between these two compounds is the additional chlorine microbial strains suited for bioaugmentation. One example, the
atom on the latter. When 2,4-D is applied to the soil, it disappears bacterium Burkholderia (Pseudomonas) cepacia, is capable of
within a period of several weeks, as a result of its degradation by growth on 2,4,5-T and has been used successfully to remove this
microbes in the soil. When 2,4,5-T is applied, however, it is often chemical from soil samples in the laboratory. However, microbes
still present more than a year later (figure 30.10). The additional that thrive under laboratory conditions may not compete well in
chlorine atom of 2,4,5-T blocks the enzyme that makes the initial natural habitats. activated sludge process, p. 736
attack on 2,4-D. Successful bioremediation may also involve controlling meta-
Most herbicides and insecticides not only are toxic to their bolic processes by manipulating the availability of O2 and specific
target, but also have deleterious effects on fish, birds, and other growth substrates. For example, anaerobic degradation of trichlo-
animals. For example, the pesticide DDT accumulates in the fat roethylene (TCE), a solvent used to clean metal parts, results in the
of predatory birds through biological magnification (table 30.1).
Small amounts of the pollutant that contaminate water are con-
centrated in minute plankton, which are eaten by minnows, accu-
mulating even more in the fish. When large birds eat the fish, the
amount of chemical in tissues is tremendously magnified. The
continuing ingestion of DDT, which accumulates in fat, results
in an ever-greater concentration of the DDT as it passes upward
through the food chain. DDT interferes with the reproductive
process of birds, leading to the production of fragile eggs, which
break before the young can hatch. Although banned in the United
States, DDT is still used in other countries, particularly to control
mosquitoes that transmit malaria.

TABLE 30.1 Biological Magnification of DDT

Parts per Million DDT Source
FIGURE 30.11 Oil Spill Bioremediation Bioremediation was
0.00005 Water used to clean the shoreline contaminated by oil spilled from the
0.04 Plankton Exxon Valdez. The growth of local oil-degrading prokaryotes was
stimulated by addition of nutrients. Comparing the uncleaned
0.23 Minnow
rocks on the left to the rocks cleaned by bioremediation on the
3.57 Heron right shows the dramatic efficiency of bioremediation.
22.8 Merganser (fish-eating duck) ? Which kind of bioremediation is this—biostimulation or
bioaugmentation?
746 Chapter 30 Environmental Microbiology

accumulation of vinyl chloride, a compound more toxic than TCE. Both nutrients and O2 may be added to facilitate microbial growth
Aerobic conditions are important to prevent this buildup. Some pol- and metabolism, while the slurry is agitated to ensure that the
lutants are degraded only when specific substrates are made available microbes remain in contact with the contaminants. A slower pro-
to the microbes. This phenomenon, called co-metabolism, occurs cess involves mounding the contaminated soil over a layer that
because the enzyme produced by the microbe to degrade the additional traps seeping chemicals. To provide O2, the soil can be turned
substrate degrades the pollutant as well. As an example, the enzymes occasionally or air forced through.
produced by some microbes to degrade methane also degrade TCE. In
this case, adding methane enhances the degradation of TCE. MicroAssessment 30.4
Bioremediation may be done either in situ (“in place”) or Bioremediation uses microorganisms to degrade pollutants.
off-site. In situ bioremediation generally relies on biostimulation Biostimulation and bioaugmentation are two methods employed.
and is less disruptive. Oxygen (O2) can be added to contaminated 10. Why do xenobiotics often persist in the environment?
groundwater and soil either by injecting hydrogen peroxide, 11. How is biostimulation different from bioaugmentation?
which rapidly decomposes to liberate O2 and water, or pumping
12. In treating an oil spill, why might biostimulation be
air into soil. Off-site processes may be performed using a bio- preferred over bioaugmentation? +
reactor, a large tank designed to accelerate microbial processes.

FUTURE CHALLENGES 30.1

Better Identification of Pathogens in Water and Wastes
One of the most important challenges in the Cysts of Giardia and oocysts of Viruses can also be detected by PCR. The
field of water and waste treatment is the devel- Cryptosporidium have been detected by ampli- viruses are concentrated by filtration onto
opment of new and better methods to detect fying specific regions of their DNA by PCR. membranes. Many different viruses can be
waterborne contaminants in both drinking In fact, using this method, it is possible to detected simultaneously by combining gene
water and environmental water samples. This detect a single cyst of Giardia and to distin- probes from various groups of viruses. A
would make it possible to follow the occur- guish between species that are pathogenic for problem is that viruses inactivated by disin-
rence and persistence of pathogens in water humans and those that are not. But problems fection procedures are still detected by PCR.
supplies with greater accuracy, and aid in bet- arise in using these techniques with environ- To overcome this, viruses can be put into cell
ter reporting of waterborne illnesses. Methods mental samples that contain substances that cultures to allow them to replicate, indicating
being developed, but not yet perfected for use inhibit the reaction. In addition, PCR detects that they are not inactivated, and the PCR is
in this field, include the polymerase chain DNA from dead organisms as well as living. then performed on the infected cell cultures.
reaction (PCR) and a variety of fluorescence Studies are needed to make these techniques
techniques and radioactivity labeling methods. feasible for use in water testing.

Summary
30.1 ■ Microbiology of Wastewater Treatment Water Testing

Biochemical Oxygen Demand (BOD) Total coliforms and fecal coliforms are used as indicator organisms;
their presence suggests the possible presence of pathogens.
An important goal of wastewater treatment is the reduction of the
biochemical oxygen demand (BOD).
30.3 ■ Microbiology of Solid Waste Treatment
Municipal Wastewater Treatment Methods (figures 30.1–30.3)
Sanitary Landfills for Solid Waste Disposal
Primary treatment is a physical process designed to remove materials
that sediment out. Secondary treatment is chiefly a biological process Landfills are used to dispose of solid wastes near towns and cities.
designed to convert most of the suspended solids to inorganic com- Municipal and Backyard Composting—Alternative
pounds and microbial biomass, removing most of the BOD. Advanced to Landfills
treatment is often designed to remove ammonia, nitrates, and phos-
Composting reduces the amount of garbage sent to landfills
phates. Biosolids that result from anaerobic digestion of sludge can be
(figures 30.8, 30.9).
used to improve soils and promote plant growth.
Individual Wastewater Treatment Systems 30.4 ■ Microbiology of Bioremediation
Rural dwellings often use septic systems for wastewater treatment
(figure 30.4). Pollutants
Synthetic compounds are more likely to be biodegradable if they have a
30.2 ■ Drinking Water Treatment and Testing chemical composition similar to that of naturally occurring compounds
(figure 30.10).
Water Treatment Processes
Community drinking water is treated to remove particulate and sus- Means of Bioremediation
pended matter, various microorganisms, and organic chemicals (figure 30.6). Biostimulation can be used to increase the effectiveness of oil degrada-
Chlorine or other disinfectants are then used to destroy harmful microbes. tion by naturally occurring prokaryotes (figure 30.11).
 Part V Applied Microbiology 747

Review Questions
Short Answer c) The media used test for the ability to ferment lactose.
1. Describe how the BOD of a water sample is determined. d) A positive test indicates that pathogens are definitely present in
2. Which step of wastewater treatment removes most of the BOD? the sample.
e) All coliforms hydrolyze ONPG and MUG.
3. Compare and contrast the activated sludge process and the trick-
ling filter system used in secondary treatment of wastewater. 8. Landfills are often used to dispose of
4. Why is it beneficial to remove nitrates and phosphates in wastewater? a) household wastewater. b) commercial wastewater.
5. How does a septic system work? c) solid wastes. d) petroleum wastes.
6. What is an aquifer? e) wastewater effluent.
7. Why do water-testing procedures look for coliforms rather than 9. Backyard composting is an excellent way to dispose of
pathogens? a) cooking fats. b) garden debris. c) spoiled meats.
8. How does the ONPG/MUG test allow a sample to be assayed d) insecticides. e) cleaning supplies.
simultaneously for the presence of both total coliforms and E. coli? 10. Synthetic compounds are most likely to be biodegradable
9. What aspect of 2,4,5-T makes it more likely to persist in the envi- if they
ronment than 2,4-D? a) are totally different from anything found in nature.
10. Describe the use of bioremediation in the cleanup of oil spills. b) have three chlorine atoms per molecule.
c) are plastics.
Multiple Choice d) are present in very large amounts.
1. A marked decrease in BOD during secondary treatment indicates e) are chemically similar to naturally occurring substances.
a) lack of oxidation during treatment.
b) effective aerobic decomposition during treatment. Applications
c) effective anaerobic decomposition during treatment. 1. A developer is interested in building vacation homes on 150 acres
d) removal of all pathogenic bacteria. of oceanfront property. A priority is to retain as much natural
e) removal of all toxic chemicals. beauty of the area as possible. Safe and effective wastewater treat-
2. Advanced treatment is often designed to remove ment must be part of the plan. What advantages and disadvantages
of each of the following options must the developer consider
a) BOD. b) nitrates and phosphates. c) bacteria.
before selecting one?
d) protozoa. e) methane.
a) Individual septic systems for each home
3. Which of the following is not a matching pair?
b) Trickling filter system
a) Potable water—presence of pathogens
c) Constructed wetlands
b) High BOD—high organic content
2. A public health official is investigating waterborne diseases in
c) Stabilized sludge—fertilizer
Illinois. She notes that over half of the cases of waterborne diseases
d) Primary treatment—removal of material that settles
originating from drinking water were caused by Giardia lamblia.
e) Bulking—growth of filamentous bacteria Other data showed that most cases of gastroenteritis attributed to
4. Which of the following is false? exposure to recreational waters were caused by Cryptosporidium
a) Bulking interferes with trickling filter systems. parvum. What does this suggest about controlling waterborne
b) Artificial wetlands provide a habitat for wildlife. diseases?
c) Removal of nitrates by microorganisms requires anaerobic
conditions. Critical Thinking +
d) Methane is a by-product of anaerobic digestion. 1. Why is oil not degraded when in a natural habitat underground yet
5. Which of the following is not a matching pair? is susceptible to bioremediation in an oil spill?
a) Surface water—watershed 2. The accompanying figure shows the effects of different treatments
b) Groundwater—aquifer of drinking water on the incidence of typhoid fever in Philadelphia,
1890–1935. If filtration of drinking water caused such a dramatic
c) Sand and gravel filters—removes organic chemicals
decrease in the disease incidence, was it necessary to introduce
d) Alum—causes suspended material to coagulate
chlorination a few years later? Why or why not?
e) Disinfection—chlorine, ozone, or ultraviolet light
6. Septic tanks should be placed
Number of typhoid cases
per 10,000 population

a) as close to the well as possible. 700

Filtration of
b) at least 500 feet from the house. 600 drinking water
c) under the house. 500
d) in deep clay soil. 400
300 Chlorination of
e) where the outflow cannot contaminate any water supply. drinking water started
200
7. Which of the following about coliform testing methods is true?
100
a) All determine the number of E. coli present in a sample. 0
1890 1900 1910 1920 1930
b) The MPN procedure precisely indicates the concentration of
Year
coliforms.
31 Food Microbiology
KKE EYYT TE ERRMMS S
Fermented Foods Foods
intentionally altered during
production by encouraging the
activity of bacteria, yeasts, or molds.
product contaminated with microbes
that colonize the host to cause
disease.
Foodborne Intoxication An
Food Preservation Increasing illness that results from consuming
the shelf life of foods by preventing an exotoxin produced by a
the growth and activities of microorganism growing in a food
microorganisms. product.
Food Spoilage Undesirable Water Activity (aw) The relative
biochemical changes in foods. amount of water available for
microbial growth; pure water has an
Foodborne Infection An illness
aw of 1.0.
that results from consuming a food

an ecosystem. Microbes compete for available nutrients, and the

most successful ones predominate.
Microorganisms on foods are not necessarily undesirable.
Sometimes, their growth results in pleasant flavors or textures.
Refrigeration is a method of food preservation. Foods intentionally altered during production by carefully
controlling the activity of bacteria, yeasts, or molds are called
fermented foods (figure  31.1). For example, food manufac-
turers purposely encourage certain microorganisms to grow in
A Glimpse of History milk in order to produce foods such as sour cream and cheese.
Alice Catherine Evans, the first female president of the Society of Alcoholic beverages, such as beer and wine, and many Asian
American Bacteriology (now the American Society for Microbiology), food condiments, such as soy sauce and miso, also rely on
helped show that unpasteurized milk could be a source of disease in microbial metabolism for their production. Strictly speaking, the
humans. A graduate of both Cornell University and the University of
term fermentation refers to only those metabolic activities that
Wisconsin, Evans worked for the U.S. Department of Agriculture, seek-
use pyruvate or another organic compound as an electron accep-
ing out the sources of microbial contamination of dairy products. In 1917,
Evans reported that cases of human brucellosis were related to finding tor, with the result that alcohols and acids are produced. Food
Brucella abortus in cows’ milk. brucellosis, p. 676
scientists, however, use the term more generally, to encompass
Evans’s conclusion that B. abortus could be transmitted from cows any desirable change that a microorganism imparts to food.
to humans through milk conflicted with the common view of a number of fermentation, p. 147
prominent scientists, including Robert Koch. In 1900, Koch had declared Undesirable biochemical changes in foods are called spoil-
that bovine tuberculosis and brucellosis could not be transmitted to age (figure  31.2). The processes that cause spoilage are often
humans. As a result, many scientists and dairy workers would not accept the same ones involved in fermentation of foods. In fact, a food
the increasing evidence that diseases were being transmitted from cows product considered by one cultural population to be fermented
to humans through milk. In the late 1930s, after a number of children of may be considered spoiled by another. Sour milk and moldy bread
dairy workers had died of brucellosis, the problem was finally acknowl- are examples of foods considered spoiled as a result of microbial
edged. Today, milk is routinely pasteurized, and only very small amounts
growth. The souring of milk, however, involves the same micro-
of unpasteurized milk are sold in the United States.
bial processes that cause the agreeable acidic flavor of sour cream,
and the mold growing on bread may be related to the one that

W
hen you prepare a meal, you also invite microorgan- causes the blue veining in Gorgonzola cheese.
isms to dinner. Practically all the food we purchase Growth of pathogens in food can result in foodborne illness,
or grow—fruit, vegetable, meat, or dairy product— but generally does not cause noticeable changes in food quality.
harbors a variety of microbes. This is not surprising considering Depending on the type of pathogen, the illness may result from
that bacteria and fungi are ubiquitous and especially plentiful in consuming either the living organisms or the toxins they have
soil and around animals. From a microbial perspective, food is produced during growth.

748
 Part V Applied Microbiology 749

31.1 ■ Factors Influencing the

Growth of Microorganisms
in Foods
Learning Outcome
1. Describe five intrinsic factors and two extrinsic factors that
influence the growth of microorganisms in foods.

Understanding the factors that influence microbial growth is

essential to maintaining food quality when producing fermented
foods or prolonging the shelf life of perishable foods. As a general
rule, bacteria will predominate in fresh meats and other moist,
pH-neutral, nutrient-rich foods. Yeasts and molds can also grow
in these foods, but the more rapid increase of bacteria overwhelms
the competitors. When conditions such as lack of moisture or high
acidity restrict the growth of bacteria, fungi predominate despite
their relatively slow growth.

Intrinsic Factors
The inherent conditions in the food—such as water availability,
acidity, and nutrient level—are called intrinsic factors. They
FIGURE 31.1 Fermented Foods affect which microbes predominate on the product.
? What makes a food “fermented”?
Water Availability
Foods vary in terms of how much water is accessible to microor-
Suppressing or limiting microbial growth can preserve the ganisms. Fresh meats and milk, for example, have plenty of water
quality of foods and prevent foodborne illnesses. Foods can be to support the growth of many microbes. Bread, nuts, and dried
canned, pasteurized, or irradiated to eliminate or decrease the foods, on the other hand, are relatively dry. Jams, jellies, and some
numbers of microorganisms. Alternatively, microbial growth can other sugar-rich foods are seemingly moist, but most of that water
be inhibited by storing food at cold temperatures, or by adding is chemically interacting with the sugar, making it unavailable for
preservatives (growth-inhibiting ingredients). The end products of use by microbes. Highly salted foods, for similar reasons, have
some fermentation processes can preserve food by preventing the little available moisture.
growth of many undesirable microorganisms. The term water activity (aw) is used to indicate the amount
of water available in foods. By definition, pure water has an aw of
1.0. Most fresh foods have an aw above 0.98, whereas ham has an
aw of 0.91, jam has an aw of 0.85, and some cakes have
FIGURE 31.2 Examples of Spoiled Food an aw of 0.70. Most bacteria require an aw above 0.90
(a) Rotten apples. (b) Moldy bread. (c) Ear of corn and for growth, which explains why fresh, moist foods
a lemon after several weeks in the refrigerator.
spoil more quickly than dried, sugary, or salted
? How is a spoiled food different from a fermented foods. Fungi can grow at an aw as low as 0.80, so
food?

(a) (b) (c)

750 Chapter 31 Food Microbiology

forgotten bread, cheese, jam, and dried foods often become moldy. Extrinsic Factors
Staphylococcus sp., which are adapted to grow on the dry, salty
Environmental conditions, such as the storage temperature and
surfaces of skin, can grow at an aw of 0.86, lower than the mini-
atmosphere, are called extrinsic factors. The extent of microbial
mum required by most common spoilage bacteria. Staphylococcus
growth varies greatly, depending on the conditions under which a
sp. normally do not compete well with other bacteria, but on salty
food is stored. Microorganisms multiply rapidly in warm, O2-rich
products such as ham and other cured meats, they can multiply
environments such as meats stored at room temperature.
with little competition. Ham is a common vehicle for S. aureus
food poisoning. Staphylococcus aureus foodborne illness, p. 393
Storage Temperature
The storage temperature affects the growth rate of microorgan-
pH isms. At low temperatures above freezing, many enzymatic reac-
Many bacterial species, including most pathogens, are inhibited by tions are either very slow or non-existent, with the result that
acidic conditions and cannot grow at a pH below 4.5. An excep- microorganisms multiply slowly, if at all. Microorganisms that
tion is the lactic acid bacteria, which can grow at a pH as low as grow on refrigerated foods are most likely psychrophiles or psy-
3.5. These bacteria produce lactic acid as a result of fermentative chrotrophs, such as some members of the genus Pseudomonas. At
metabolism and are used to make yogurt, sauerkraut, and some freezing temperatures, water becomes crystalline and inaccessible,
other fermented foods. They are also prime causes of spoilage of stopping microbial growth. psychrophiles, p. 89 psychrotroph, p. 89
unpasteurized milk and other foods. lactic acid bacteria, p. 258
Fungi can grow at a lower pH than most spoilage bacteria, Atmosphere
so some acidic foods eventually become moldy. For example, the The presence or absence of O2 affects the type of microbial popu-
pH of lemons is approximately 2.2, which inhibits the growth of lation able to grow in food. Obligate aerobes cannot grow in foods
bacteria, including the lactic acid group. However, some fungi can stored under conditions that exclude their required O2. Keeping O2
grow at this low pH. out of food, however, may permit other bacteria to grow, includ-
The pH of a food product can also determine whether tox- ing the obligate anaerobe Clostridium botulinum. A case of botu-
ins can be produced. Clostridium botulinum (the bacterium that lism was traced to the consumption of a thick, homemade stew
causes botulism) does not grow or produce toxin below pH 4.5, that had been slowly cooked and then left at room temperature
so it is not considered a danger in highly acidic foods. This is overnight. The cooking process did not destroy the endospores of
why the canning process for acidic fruits and pickles is less strin- C. botulinum and had driven off the O2, thereby creating anaero-
gent than that for foods with a higher pH. Some newer varieties bic conditions in which the organism germinated, multiplied, and
of tomatoes are less acidic than older types, requiring that acid produced toxin. oxygen requirements, p. 90
be added if they are to be safely canned using the less stringent
procedures. MicroAssessment 31.1
Intrinsic factors (such as available moisture, pH, and the presence
Nutrients of antimicrobial chemicals) and extrinsic factors (including storage
An organism requiring a particular vitamin cannot grow in a temperature and atmosphere) influence the type of microorganisms
that grow and predominate in a food product.
food lacking that vitamin. A microbe capable of synthesizing that
vitamin, however, can grow if other conditions are favorable. 1. Why is Staphylococcus aureus more likely to be found in high
Members of the genus Pseudomonas often spoil foods because numbers on ham than on fresh meat?
they can synthesize essential nutrients and can multiply in various 2. Which is important to refrigerate: fresh stew or bread? Why?
environments, including refrigeration. 3. Why would the cooking process create anaerobic conditions? +

Biological Barriers
Rinds, shells, and other coverings help protect foods from inva- 31.2 ■ Microorganisms in Food
sion by microorganisms. Eggs, for example, retain their quality
much longer with intact shells. Whole lemons keep longer than and Beverage Production
slices. Even so, microorganisms will eventually break down these
Learning Outcomes
coverings and cause spoilage.
2. Explain why lactic acid bacteria are important in food and
beverage production.
Antimicrobial Chemicals
3. Compare and contrast the production of cheese, yogurt, and
Some foods contain natural antimicrobial chemicals that help acidophilus milk.
prevent spoilage. Egg white, for instance, is rich in lysozyme. 4. Describe the production of pickled vegetables and fermented meat
If lysozyme-susceptible bacteria breach the protective shell of products.
an egg, they are destroyed by lysozyme before they can cause 5. Compare and contrast the production of wine, beer, distilled
spoilage. Other examples of naturally occurring antimicrobial spirits, and vinegar.
chemicals are benzoic acid in cranberries and allicin in garlic. 6. Describe the production of soy sauce.
lysozyme, p. 62
 Part V Applied Microbiology 751

Fermented foods, such as yogurt, cheese, and pickled vegetables, Cheese, Yogurt, and Other
are perceived as pleasant tasting. In addition, their acids inhibit the Fermented Milk Products
growth of many spoilage organisms as well as foodborne patho- Milk is sterile in a cow’s udder, but rapidly becomes contaminated
gens. Thus, fermentation historically has been, and continues to with a variety of microorganisms during milking and handling.
be today, an important method of food preservation, particularly Various species of lactic acid bacteria are inevitably introduced
when modern conveniences such as refrigeration are lacking. because they commonly reside on the udder. If the milk is not
refrigerated, these bacteria grow and ferment lactose—the main
Lactic Acid Fermentations sugar in milk—producing lactic acid. The combined effect of
removing the primary carbohydrate as a nutrient source and accu-
by the Lactic Acid Bacteria mulating lactic acid inhibits the growth of many other microbes.
The tart taste of yogurt, pickles, sharp cheeses, and some Aesthetic features of the milk change as well because lactic acid
sausages is due to the production of lactic acid by one or lowers the pH, which in turn causes the milk proteins to coagulate
more members of a group of bacteria known as the lactic acid or curdle, and sours the flavor. milk spoilage, p. 721
bacteria (table  31.1). These bacteria—including species of Today, with high quality control standards and the use of pas-
Lactobacillus, Lactococcus, Streptococcus, Leuconostoc, and teurized milk, the commercial production of fermented milk prod-
Pediococcus—are obligate fermenters that characteristically ucts does not rely on naturally present lactic acid bacteria. Instead,
produce lactic acid as an end product of their metabolism. starter cultures containing one or more strains of lactic acid
Some also produce flavorful and aromatic compounds that bacteria are added to the milk. These strains are carefully selected
contribute to the overall quality of fermented foods. lactic to produce the most desirable flavors and textures. Precious starter
acid bacteria, p. 258 obligate fermenters, p. 90 cultures must be carefully maintained and protected against con-
tamination, particularly by bacteriophages, which can damage or
destroy them. bacteriophage, p. 304
Cheese Cottage cheese is one of the simplest cheeses to make.
Foods Produced Using Lactic Pasteurized milk is inoculated with a starter culture, usually con-
TABLE 31.1
Acid Bacteria taining Lactococcus cremoris and L. lactis, and then incubated
Food Characteristic until fermentation products cause the proteins in milk to coagu-
late. The coagulated proteins, or curd, are heated and cut into
Milk Products
small pieces to make it easier to drain the liquid waste portion.
Cheese Uses a starter culture usually containing Unlike most cheeses that undergo further microbial processes
(unripened) Lactococcus cremoris and L. lactis
called ripening or curing, cottage cheese is unripened.
Cheese (ripened) Uses rennin and a starter culture The initial steps of ripened cheese production are the same
containing Lactococcus cremoris and
L. lactis; ripened for weeks to years;
as those of cottage cheese, except the enzyme rennin is added to
other bacteria and/or fungi may be the fermenting milk to speed protein coagulation (figure  31.3).
added to enhance flavor development After the proteins coagulate, the whey (the liquid waste portion)
Yogurt Uses a starter culture containing is removed. The curds are then salted, pressed, and shaped into
Streptococcus thermophilus and the traditional forms, usually bricks or wheels. The cheese is then
Lactobacillus delbrueckii subspecies ripened, resulting in characteristic textural and flavor changes due
bulgaricus to the metabolic activities of naturally occurring or starter lactic
Sweet acidophilus Lactobacillus acidophilus added for acid bacteria. Depending on the type of cheese, ripening can take
milk possible health benefits from several weeks to years. Longer ripening creates more acidic,
Vegetables sharper cheeses.
Sauerkraut Cabbage; succession of naturally Some cheeses are inoculated with other bacteria or fungi that
occurring bacteria including Leuconostoc give characteristics particular to the kind of cheese. For example,
mesenteroides, Lactobacillus brevis, and the bacterium Propionibacterium shermanii ripens Swiss cheese
Lactobacillus plantarum
and gives it the characteristic holes and a nutty flavor. This
Pickles Cucumbers; naturally occurring bacteria bacterium ferments organic compounds to produce propionic acid
Poi Taro root; naturally occurring bacteria; and CO2. The CO2 gas causes the holes in the cheese, while the
Hawaii propionic acid gives the typical flavor. Propionic acid also inhibits
Olives Green olives spoilage organisms. Roquefort, Gorgonzola, and Stilton cheeses are
Kimchee Cabbage and other vegetables; Korea ripened by the fungus Penicillium roquefortii. Growth of the fungus
Meats along cracks in the cheese gives these cheeses the distinctive bluish-
green veins. Brie and Camembert are ripened by a white fungus
Dry and semidry Uses a starter culture containing species
sausages of Lactobacillus and Pediococcus; meat is such as P. candidum or P. camemberti inoculated on the surface of
stuffed into casings, incubated, heated, the cheese. As the fungal cells grow into the cheese, they produce
and then dried enzymes that alter texture and flavor. Limburger cheese is made in
a similar manner, but with the bacterium Brevibacterium linens.
752 Chapter 31 Food Microbiology

is added immediately before packaging. The bacteria are simply

included for their possible health benefits. Some evidence sug-
gests they may aid in the digestion of lactose as well as prevent
and reduce the severity of some diarrheal illnesses, but the role
they play in the complex interactions of the human intestinal tract
is not clear. Unlike most lactic acid bacteria used as starter cul-
tures, L. acidophilus can potentially colonize the intestinal tract.

MicroByte
The enzyme rennin is found in calves’ stomachs, where it helps digest
the mother’s milk. Today, genetically engineered microbes make it.
Coagulation— Lactic acid production and rennin activity
cause the milk proteins to coagulate. The coagulated
mixture is then cut so that the liquid whey will start
separating from the solid curd. Pickled Vegetables
Another fermentation process known as pickling originated as
a way to preserve vegetables such as cucumbers and cabbage.
Today, pickled products such as sauerkraut (cabbage), pickles
(cucumbers), and olives are valued for their flavor. Fermentation
of most vegetables uses naturally occurring lactic acid bacteria
from the vegetables rather than starter cultures.
One of the most well-studied natural fermentations is the pro-
duction of sauerkraut. The cabbage is first shredded and layered
with salt. The layers are firmly packed to provide an anaerobic
environment. The salt draws water and nutrients from the cab-
bage, creating a brine that inhibits most microbes other than the
Separation of curds from whey— The curd is heated lactic acid bacteria. Under the correct conditions, natural succes-
and cut into small pieces. The liquid whey is removed
by draining.
sions of lactic acid bacteria grow. These bacteria—Leuconostoc
mesenteroides, Lactobacillus brevis, and Lactobacillus
plantarum—produce lactic acid, which lowers the pH, further
inhibiting undesired microbes. The lactic acid and other fermen-
tation end products give sauerkraut its characteristic tangy taste.
When the desired flavor has developed, usually after 2 to 4 weeks at
room temperature, the sauerkraut is often canned. Similar processes
are used to make some pickles, olives, and other vegetable products.

Fermented Meat Products

Fermented meat products—such as salami, pepperoni, and sum-
mer sausage—were traditionally produced by allowing the small
Aging— Curds are salted and pressed into blocks or numbers of lactic acid bacteria naturally present to multiply to
wheels for aging. the point of dominance. Relying on the natural fermentation of
FIGURE 31.3 Commercial Production of Cheese meat is inherently risky, however, because the incubation condi-
? Why does a longer ripening process give rise to a sharper cheese?
tions can potentially support the growth and toxin production
of pathogens such as Staphylococcus aureus and Clostridium
botulinum. Starter cultures are now used because they ensure that
Yogurt To produce yogurt, pasteurized milk is concentrated lactic acid is rapidly produced, thereby inhibiting the growth of
slightly by evaporation and then inoculated with a starter cul- pathogens and improving flavor development. Starter cultures
ture containing Streptococcus thermophilus and Lactobacillus used by U.S. sausage-makers typically contain Lactobacillus
delbrueckii subspecies bulgaricus. The mixture is incubated at and/or Pediococcus species.
40°C to 45°C for several hours, during which time these thermo- To make fermented sausages, meat is ground and combined
philic bacteria grow rapidly. They produce lactic acid and other with a starter culture and other ingredients including sugar, salt,
end products that contribute to the flavor. Carefully controlled and nitrite. The sugar serves as a substrate for fermentation,
incubation conditions favoring the balanced growth of the two because meat does not naturally contain enough fermentable
species ensure the proper levels of acid and flavor compounds. carbohydrate to produce adequate lactic acid. Salt and nitrite
contribute to the flavor and also inhibit the growth of spoilage
Acidophilus Milk Traditional acidophilus milk is the product microorganisms. More importantly, they inhibit C. botulinum.
of fermentation by Lactobacillus acidophilus. The more readily After thorough blending, the mixture is stuffed into a casing and
available sweet acidophilus milk retains the flavor of fresh milk incubated from one to several days. The product can then be
because it is not fermented. Instead, a culture of L. acidophilus smoked or otherwise heated to kill bacteria. Finally, it is dried.
 Part V Applied Microbiology 753

Alcoholic Fermentations by Yeast

Some yeasts, such as members of the genus Saccharomyces,
1 Crushing
ferment simple sugars to produce ethanol and CO2. They are
Grapes are crushed, and the
used to make alcoholic beverages as well as vinegar and bread resulting must (solids and juice)
(table 31.2). is collected.

Wine
Wine is the product of the alcoholic fermentation of naturally
occurring sugars in the juices of grapes or other fruits. One of the Yeast
most important variables in wine is the variety and quality of fruit
used. The growing conditions and ripeness as well as other factors
2 Fermentation
affect the content of sugar, acids, and various organic compounds,
A specially selected strain of
which in turn critically influences the final product. Saccharomyces cerevisiae is added
The commercial production of wine begins by crushing to ferment the must (red wines) or
grapes in a machine that removes the stems and collects the result- clear juices (white wines).
ing solids and juices, or must (figure 31.4). If white wine is to be
made, only the clear juices are fermented. For red wine, the entire
must of red grapes is put into the fermentation vat. The color and
complex flavors of these wines are derived from components of 3 Settling
the grape skin and seeds. The solids are removed during fermenta- The fermented material is transferred
tion once the desired amount of color and flavor compounds have to a settling tank so that the solids
can be removed.
been extracted. Rose wines obtain their light pink color from the
entire crushed red grape fermented for about 1 day, after which the
juice is removed and fermented alone.
Fermentation starts when a specially selected strain of Solids
Saccharomyces cerevisiae is inoculated. Sulfur dioxide (SO2) is
generally added to inhibit the growth of the natural microbial pop-
ulation of the grape, especially acetic acid bacteria. These bacteria 4 Aging
Red wines and some white wines are
convert alcohol to acetic acid (vinegar) and are a common cause aged in oak barrels.
of wine spoilage. The S. cerevisiae strains used to make wine are

Foods and Beverages Produced 5 Bottling

TABLE 31.2 Using Alcoholic Fermentation The wine is clarified by filtration and
by Yeast then bottled.

Product Characteristic

Alcoholic Beverages FIGURE 31.4 Commercial Production of Wine

Wine Sugars in grape juice are fermented by ? How is the initial step of making white wines different from that
Saccharomyces cerevisiae. of making red wines?
Sake Amylase from mold (Aspergillus oryzae)
converts the starch in rice to sugar,
which is then fermented by S. cerevisiae. more resistant to the antimicrobial action of SO2 and produce a
Beer Enzymes in germinated barley convert higher alcohol content than naturally occurring yeasts.
starches of barley and other grains Fermentation is carried out at a carefully controlled tem-
to sugar, which is then fermented by
perature, which varies with the type of wine, for a period ranging
S. cerevisiae.
from a few days to several weeks. During fermentation most of
Distilled spirits Sugars, or starches that are converted to
the sugar is converted to ethanol and CO2, generally resulting in a
sugars, are fermented by S. cerevisiae;
distillation purifies the alcohol. final alcohol content of less than 14%. Dry wines result from the
complete fermentation of the sugar, whereas sweet wines contain
Vinegar Alcohol produced by fermentation is
oxidized to acetic acid by species of residual sugar.
Gluconobacter or Acetobacter. In addition to the alcoholic fermentation of the grape sugars,
Breads S. cerevisiae ferments sugar; expansion a distinctly different type of fermentation, called malolactic fer-
of CO2 causes the bread to rise; alcohol mentation, can occur during wine production. Lactic acid bacteria,
evaporates during baking. primarily species of Leuconostoc, convert malic acid to the less
acidic lactic acid. Red wines made from grapes grown in cool
754 Chapter 31 Food Microbiology

regions tend to have high levels of malic acid, and their flavor is
mellowed by this fermentation.
After fermentation, the wine is transferred to a settling tank, 1 Milling
where fermented solids can settle out. Most red wines and some Malt (germinated barley) is cracked
white wines are then aged in oak barrels, contributing to the com- open.
plexity of the flavor. Finally, wine is clarified by filtration and Mill
bottled. The CO2 produced during fermentation is usually released
before the wine is bottled, resulting in a “still” (non-carbonated)
wine. Other processes are used to prepare carbonated wines such
as champagne. 2 Mashing
The ingredients of mash—malt, water,
The Japanese wine sake depends on several microbial fer- and sometimes adjuncts—are mixed.
mentation reactions. First, cooked rice is inoculated with the Enzymes in the mash convert
fungus Aspergillus oryzae. The fungus produces the enzyme starches into fermentable sugars.
Mash bin Spent grains are then removed.
amylase, which degrades the rice starch to sugar. Then, a strain of
Hops
Saccharomyces cerevisiae is added to convert the sugar to alcohol
and CO2. Lactic acid bacteria add to the flavor by producing lactic Spent grains
acid and other fermentation end products.
3 Boiling
Hops are added and the mixture is
Beer boiled to extract the flavor, concentrate
Beer production is a multistep process designed to break down the wort, inactivate enzymes, kill most
the starches of grains such as barley to produce simple sugars, microbes, and precipitate proteins.
Brew kettle
Spent hops are then removed.
which are then fermented by yeast (figure  31.5). Yeasts alone
cannot convert grain to alcohol because they lack the enzymes Wort
Yeast Spent hops
that degrade starch, the primary carbohydrate of grain. Sprouted
or germinated barley, however, known as malted barley or malt,
naturally contains these and other important enzymes. The malt is
dried and milled (ground) in preparation for the brewing process. 4 Fermentation
In a process called mashing, the malt is mixed with adjuncts Yeast is added. When fermentation is
(starches, sugars, or whole grains such as rice, corn, or sorghum), complete, excess yeast cells are then
Fermenting
and then soaked in warm water. During mashing, enzymes of the removed.
tank
malt act on the starches, converting them to fermentable sugars.
The final characteristics of the beer, such as color, flavor, and
Yeast
foam, are derived entirely from compounds in the malt. The
adjuncts simply serve as less expensive sources of carbohydrates 5 Aging
for alcohol production. The beer is aged, allowing time for
After mashing, the spent grains (residual solids) are removed flavors to develop.
Lagering tank
to yield the sugary liquid called wort. Hops, the flowers of the
vinelike hop plant, are added to the wort to impart a desirable
bitter flavor to the beer and contribute antibacterial substances. 6 Bottling
The mixture is boiled to extract the flavor components of hops, The beer is filtered or pasteurized and
concentrate the wort, inactivate enzymes, kill most microbes, and then bottled.
precipitate proteins, facilitating their removal. The wort is then
centrifuged to remove the solids and cooled before being trans- FIGURE 31.5 Commercial Production of Beer
ferred to the fermentation tank. ? How are the yeasts used to make ales different from the ones
Special strains of brewer’s yeasts (as opposed to baker’s used to make lagers?
yeasts) are commonly used in beer-making. A group of brewer’s
yeasts, called “bottom yeasts”—which includes a Saccharomyces The fermentation process generates beer with an alcohol
cerevisiae strain referred to as S. carlsbergensis—tend to form content ranging from 3.4% to 6%. Most of the yeasts settle out
clumps that sink to the bottom of the fermentation vat. These following fermentation and are removed. These yeasts can be sold
yeasts ferment best at temperatures between 6°C and 12°C and as flavor and dietary supplements. The beer is then aged, during
usually take 8 to 14 days to complete fermentation. In contrast, which time residual, unwanted flavor compounds are metabo-
“top-fermenting yeasts”—other strains of S. cerevisiae—are lized by remaining yeast cells or settle out. Cask-conditioned
distributed throughout the wort but are carried to the top of the beer undergoes a second fermentation, which generates CO2 in
vat by the rising CO2. They ferment at higher temperatures (14°C the cask. Other beers must be carbonated to replace the CO2 that
to 23°C) and over a shorter period (5 to 7 days). Most American escapes during fermentation. After aging, beer is clarified by
beers are lagers produced by the bottom yeasts. Ales, porters, and filtration, microorganisms are removed or killed using membrane
stouts are made using top-fermenting yeasts. filtration or pasteurization, and the product is packaged.
 Part V Applied Microbiology 755

Distilled Spirits FIGURE 31.6 Bread

The manufacturing of distilled spirits such as scotch, whiskey, and ? What creates the air
gin is initially similar to that of beer, except the wort is not boiled. pockets in bread?
Consequently, enzymes in the wort continue breaking down the
starches during fermentation. When fermentation is complete, the
ethanol is collected by distillation.
Different types of spirits are made in different ways. For
example, rum is made by fermenting sugar cane or molasses. Malt
scotch whiskey is the product of the fermentation of barley that is
then aged for several years in oak sherry casks. The wood and the
residual sherry contribute both flavor and color to the whiskey as
it ages. Lactic acid bacteria are used to produce lactic acid in grain
mash for making sour-mash whiskey. The yeast S. cerevisiae sub-
sequently ferments the sour mash to form alcohol. The distilled
spirit tequila is traditionally made from the fermentation of juices
from the agave plant using the bacterium Zymomonas mobilis.
This bacterium ferments sugars to ethanol and CO2 via a pathway
similar to the yeast alcoholic fermentation pathway.

Vinegar
Vinegar, an aqueous solution of at least 4% acetic acid, is Changes Due to Mold Growth
the product of the oxidation of ethanol by the acetic acid
Molds contribute to the flavor and texture of some cheeses, as
bacteria—Acetobacter and Gluconobacter species. Acetic acid
already discussed. In addition, many traditional dishes and condi-
bacteria are strictly aerobic, Gram-negative rods, character-
ments used throughout the world are produced by encouraging the
ized by their ability to carry out a number of oxidations. They
growth of molds on food (table  31.3). Successions of naturally
can tolerate high concentrations of acid as they oxidize alcohol
occurring microorganisms are often involved. The microbiologi-
to acetic acid.
cal and chemical aspects of many of these foods have not been
Alcohol is commercially converted to vinegar using pro-
extensively studied.
cesses that provide O2 to speed the oxidation reaction. One
method uses a vinegar generator, which sprays alcohol onto
Soy Sauce
loosely packed wood shavings that harbor a biofilm of acetic
acid bacteria. As the alcohol trickles through the bacteria-coated Soy sauce is made by inoculating equal parts of cooked soybeans
shavings, it is oxidized to acetic acid. In principle, the vinegar and roasted cracked wheat with a culture of either Aspergillus
generator operates much like the trickling filter used in waste- oryzae or A. sojae. The mixture, called koji, is allowed to stand for
water treatment by providing a large surface area for aerobic several days, during which time carbohydrates and proteins in the
metabolism. Another method uses a submerged culture reactor,
an enclosed system that continuously pumps small air bubbles
into alcohol that has been inoculated with acetic acid bacteria.
TABLE 31.3 Foods Produced Using Molds
trickling filter, p. 738 biofilm, p. 84
Food Characteristic
Bread Soy sauce Koji is produced by inoculating soybeans
Yeast bread rises through the action of baker’s yeast—strains and cracked wheat with a starter culture of
Aspergillus oryzae or A. sojae; the mixture
of Saccharomyces cerevisiae carefully selected for the com- is then added to a brine and incubated for
mercial baking industry. The CO2 produced during fermenta- many months.
tion causes the bread to rise, producing the spongy texture Tempeh Soybeans are fermented by lactic acid bacteria
characteristic of yeast breads (figure  31.6). The alcohol and then inoculated with a species of the
evaporates during baking. mold Rhizopus; Indonesia.
Yeast bread is made from a mixture of flour, sugar, salt, milk Miso Rice, soybeans, or barley are inoculated with
or water, yeast, and sometimes butter or oil. Packaged baker’s Aspergillus oryzae; Asia.
yeast that can be reconstituted in warm water is readily available Cheeses
as pressed cakes or dried granules. An excess of yeast is added Roquefort, Curd is inoculated with Penicillium
to allow adequate production of CO2 in a time period too short to Gorgonzola, roquefortii.
permit multiplication of spoilage bacteria. and Stilton
Sourdough bread is made with a combination of yeast and Brie and Wheels of cheese are inoculated with selected
lactic acid bacteria. Lactic acid is produced, as well as alcohol and Camembert species of Penicillium.
CO2, giving the bread its sour flavor.
756 Chapter 31 Food Microbiology

soybeans are broken down, producing a yellow-green liquid con- and Lactobacillus, all produce lactic acid. Anyone who has unex-
taining fermentable sugars, peptides, and amino acids. After this, pectedly consumed sour milk knows that this can be disagree-
the mixture is put into a large container with an 18% NaCl solu- able. Bacillus and Clostridium sp. are particularly troublesome
tion (brine). Salt-tolerant microorganisms then grow and produce causes of food spoilage because their heat-resistant endospores
flavor changes over an extended period. Microorganisms involved survive cooking and, in some cases, canning. B. coagulans and
in this stage of fermentation include lactobacilli, pediococci, and B. stearothermophilus spoil some canned foods. glycocalyx, p. 62
yeasts. After the brine mixture is allowed to ferment for 8 to
12 months, the liquid soy sauce is removed. The residual solids
are used as animal feed. Common Spoilage Fungi
A wide variety of fungi, including species of Rhizopus, Alternaria,
MicroAssessment 31.2 Penicillium, Aspergillus, and Botrytis, spoil foods. Because fungi
grow readily in acidic as well as low-moisture environments,
Lactic acid bacteria are used to produce a variety of foods including
cheese, yogurt, sauerkraut, and some sausages. Yeasts are used to fruits and breads are more likely to be spoiled by fungi than by
produce alcoholic beverages and breads. Some cheeses and many bacteria. Aspergillus flavus grows on peanuts and other grains,
traditional foods owe their characteristics to changes caused by molds. producing aflatoxin, a potent carcinogen monitored by the Food
4. Describe how the metabolism of lactic acid bacteria differs from and Drug Administration.
that of most other microorganisms that can grow aerobically.
5. How does the use of starter cultures improve the safety of MicroAssessment 31.3
fermented meat products? The metabolites of microbes can spoil foods by imparting undesirable
6. How could cottage cheese be produced without bacteria? + flavors, odors, and textures.
7. What characteristics of Pseudomonas species allow them to spoil
such a wide variety of foods?
31.3 ■ Food Spoilage 8. Why do fungi most commonly spoil breads and fruits? +

Learning Outcome
7. Distinguish between fermented foods and spoiled foods. 31.4 ■ Foodborne Illness
Food spoilage encompasses any undesirable changes in food. Learning Outcome
Spoilage microorganisms produce metabolites that have undesira- 8. Distinguish between foodborne intoxication and foodborne
ble tastes and odors. Although these are aesthetically disagreeable, infection, and give two examples of each.
they are generally not harmful. This is not surprising when micro-
bial growth requirements are considered. Most human pathogens Foodborne illness, commonly referred to as food poisoning,
grow best at temperatures near 37°C, whereas most foods are occurs when a pathogen, or a toxin it produced, is consumed in a
usually stored at temperatures well below the normal body tem- food product. Food production is carefully regulated in the United
perature. Similarly, the nutrients available in fruits, vegetables, States to prevent foodborne illness. Federal, state, and local agen-
and other foods are generally not suitable for the optimum growth cies cooperate in inspections to help enforce protective laws. In
of human pathogens. As a result, the non-pathogens can easily spite of strict controls, millions of cases of food poisoning are
outgrow the pathogens when competing for the same nutrients. estimated to occur each year from foods prepared either commer-
Spoiled foods are considered unsafe to eat, however, because high cially, at home, or in institutions such as hospitals and schools.
numbers of spoilage organisms indicate that foodborne pathogens The vast majority of these cases could have been prevented with
may be present as well. proper storage, sanitation, and preparation. Table 31.4 lists some
bacteria that cause foodborne illness in the United States.
To more accurately determine the burden of foodborne ill-
Common Spoilage Bacteria ness in the United States, a government program called FoodNet
Numerous types of bacteria are important in food spoilage. (Foodborne Disease Active Surveillance Network) now collects
Pseudomonas species can degrade a wide variety of compounds, data on laboratory-confirmed cases of diarrheal illness in 10
and grow on and spoil many different kinds of foods, including states, covering approximately 15% of the population (www.cdc
meats and vegetables. Psychrotrophic species are notorious for .gov/foodnet). By gaining a better understanding of the epide-
spoiling refrigerated foods. Members of the genus Erwinia pro- miology of foodborne diseases, these diseases can hopefully be
duce enzymes that degrade pectin, and so they commonly cause prevented more easily.
soft rot of fruits and vegetables. Acetobacter species transform
ethanol to acetic acid, the principal acid of vinegar. Although
this is very beneficial to commercial producers of vinegar, it Foodborne Intoxication
presents a great problem to wine producers. Milk products are Foodborne intoxication is an illness that results from consum-
sometimes spoiled by Alcaligenes sp. that form a glycocalyx, ing an exotoxin produced by a microorganism growing in a food
causing strings of slime, or “ropiness,” in raw milk. The lactic product. It is the toxin that causes illness, not the living organisms
acid bacteria, including species of Streptococcus, Leuconostoc, (figure 31.7). Staphylococcus aureus and Clostridium botulinum
 Part V Applied Microbiology 757

TABLE 31.4 Common Foodborne Illnesses

Organism Symptoms Foods Commonly Implicated

Intoxication

Clostridium botulinum Weakness; double vision; progressive inability to speak, Low-acid canned foods such as vegetables and
swallow, and breathe meats
Staphylococcus aureus Nausea, vomiting, abdominal cramping Cured meats, creamy salads, cream-filled pastries
Infection
Campylobacter species Diarrhea, fever, abdominal pain, nausea, headache Poultry, raw milk
Clostridium perfringens Intense abdominal cramps, watery diarrhea Meats, meat products
Escherichia coli O157:H7 Severe abdominal pain, bloody diarrhea; sometimes Ground beef, raw vegetables, unpasteurized
hemolytic uremic syndrome juices
Listeria monocytogenes Influenza-like symptoms, fever, may progress to sepsis, Raw milk, cheese, meats, raw vegetables
meningitis
Salmonella species Nausea, vomiting, abdominal cramps, diarrhea, fever Poultry, eggs, milk, meat
Shigella species Abdominal cramps; diarrhea with blood, pus, or mucus; Salads, raw vegetables
fever; vomiting
Vibrio parahaemolyticus Diarrhea, abdominal cramps, nausea, vomiting, Fish and shellfish
headache, fever, chills

are two examples of bacteria that cause foodborne intoxication. compete well with most spoilage organisms, but it thrives in
exotoxin, p. 391 moist, rich foods in which other organisms have been killed or
their growth inhibited. For example, it can grow with little com-
Staphylococcus aureus petition on unrefrigerated salty products such as ham (aw of 0.91).
Many strains of Staphylococcus aureus produce a toxin that Creamy pastries and starchy salads stored at room temperature
causes nausea and vomiting when ingested. S. aureus does not also offer ideal conditions for the growth of this pathogen, because
most competing organisms were killed as
the ingredients were cooked.
Staphylococcus aureus Clostridium botulinum
The source of S. aureus is usually a
human carrier who has not followed
adequate hygiene procedures—such as
handwashing—before preparing the food.
If the organism is inoculated into a food
that supports its growth, and the food is
left at room temperature for several hours,
S. aureus can grow and produce the toxin.
Unlike most exotoxins, S. aureus toxin is
• Most bacteria that normally compete • Clostridium botulinum endospores, heat-stable, so cooking the food will not
with Staphylococcus aureus are either common in soil and marine sediments, destroy it. Staphylococcus aureus, p. 524
killed by cooking or inhibited by high contaminate many different foods.
salt conditions.
• Endospores survive inadequate
• A food handler inadvertently transfers canning processes. Canned foods Botulism
S. aureus onto food. are anaerobic. Botulism is a deadly paralytic disease caused
• S. aureus grows and produces toxin • Surviving C. botulinum endospores by ingesting a neurotoxin produced by the
when food is allowed to slowly cool or germinate, grow, and produce toxin anaerobic, spore-forming, Gram-positive rod
is stored at room temperature. in low-acid canned foods.
Clostridium botulinum (see Perspective
• A person ingests the toxin-containing • A person ingests the toxin-containing 31.1). Unfortunately, growth of the organism
food. Symptoms of staph food poisoning— food. Symptoms of botulism, including
nausea, abdominal cramping, weakness, double vision, and pro-
and toxin production may not result in any
and vomiting—begin after 4 to 6 hours. gressive inability to speak, swallow, noticeable changes in the taste or appearance
and breathe, begin in 12 to 36 hours. of the food. botulism, p. 652 neurotoxin, p. 392
Canning processes for low-acid foods
are specifically designed to destroy the endo-
FIGURE 31.7 Typical Events Leading to Foodborne Intoxications by spores of C. botulinum. If the endospores
Staphylococcus aureus and Clostridium botulinum are not destroyed or are introduced post-
? Why is botulism primarily a problem in canned foods rather than fresh ones? processing, they can germinate in these
758 Chapter 31 Food Microbiology

PERSPECTIVE 31.1
Botox for Beauty and Pain Relief
The exotoxin produced by Clostridium years the toxin, known as Botox, has become muscles in the neck and shoulders contract
botulinum is one of the most powerful poisons useful in treating various conditions. involuntarily, causing jerky movements, mus-
known. These anaerobic soil bacteria are endo- Botox is often used as a cosmetic treatment cle pain, and tremors. Injections of Botox
sporeformers found naturally on many foods. to remove facial lines, such as frown lines. directly into the affected areas give relief for
They survive usual cooking methods and inad- Extremely dilute Botox is injected directly into 3 to 4 months, after which the treatment can
equate canning procedures. Should conditions the area, paralyzing the muscles that are caus- be repeated. Another example is Parkinson’s
become anaerobic in the food, toxin can be ing the frown or other lines. Although the lines disease, a condition in which certain nerve
produced, and if ingested it causes botulism. are erased, the effect is temporary, and the treat- cells are lost, resulting in tremor, impaired
A few milligrams of this exotoxin is sufficient ment must be repeated after several months. movement, and in some cases, dystonia. Botox
to kill the entire population of a large city. The Botox is also used to relieve a num- injected into the affected muscles can give
botulinum toxin blocks transmission of acetyl- ber of very painful and disabling conditions dramatic, although temporary, relief. So, in
choline nerve signals to the muscles, resulting involving muscle contractions such as dysto- spite of its powerful and dangerous properties,
in paralysis and often in death. This is a very nia (severe muscle cramping). For example, botulinum toxin, when used with great care,
dangerous toxin. Yet surprisingly, in recent cervical dystonia is a painful disease in which can be a useful therapeutic agent.

foods. The resulting vegetative cells can then grow and produce which raw chicken was cut is then immediately used to cut
toxin. Manufacturing errors are rare in commercially canned foods, up vegetables for a salad, the salad can become contaminated
so most cases of botulism are due to improperly processed home- with Salmonella or Campylobacter species. Salmonella, p.  592
canned foods. As an added safety measure, low-acid, home-canned Campylobacter, p. 593
foods should be boiled for at least 15 minutes immediately before
serving it. The toxin is heat-labile (sensitive), so the heat treatment Escherichia coli O157:H7
will destroy any toxin that may have been produced. Escherichia coli O157:H7, a common serotype of Shiga toxin–
Cans of foods that are damaged should be discarded, because producing E. coli (STEC), causes bloody diarrhea. Infection
they might have small holes through which C. botulinum could sometimes results in hemolytic uremic syndrome (HUS), a
enter. Likewise, bulging cans indicate gas production, which life-threatening condition. hemolytic uremic syndrome, p.  598
could indicate microbial growth, and should be discarded. STEC, p. 590

Foodborne Infection Salmonella, Campylobacter E. coli O157:H7

Unlike foodborne intoxication, foodborne
infection requires the consumption of liv-
ing organisms. The symptoms of the illness,
which usually do not appear for at least 1
day after eating the contaminated food, usu-
ally include diarrhea. Thorough cooking of
food immediately before consuming it will
kill the organisms, thereby preventing infec-
tion. Escherichia coli O157:H7, Salmonella
sp., and Campylobacter sp. are examples of • Incomplete cooking fails to kill all • Incomplete cooking fails to kill all
pathogens. Surviving Salmonella and/or pathogens. Even low numbers of surviving
organisms that cause foodborne infection Campylobacter can multiply as food is E. coli O157:H7 can cause illness.
(figure 31.8). cooled slowly or stored at room
temperature. • Live organisms are ingested. They multiply
in the intestinal tract and cause disease.
Salmonella and Campylobacter • Live organisms are ingested. They multiply Symptoms include severe abdominal pain
in the intestinal tract and cause disease. and bloody diarrhea.
Salmonella and Campylobacter are two Symptoms include diarrhea, abdominal
genera commonly associated with poul- pain, and nausea.
try products such as chicken, turkey, and
eggs. Inadequate cooking of these prod-
ucts can result in foodborne infection. FIGURE 31.8 Typical Events Leading to Foodborne Infections by Salmonella,
Cross-contamination of other foods can Campylobacter, and E. coli O157:H7
result in the transfer of pathogens to those ? Why are ground meats more common than steaks as a source of E. coli O157:H7
foods. For example, if a cutting board on infection?
 Part V Applied Microbiology 759

The bacterium often colonizes the intestinal tract of healthy ■ Pasteurization. Heating foods under controlled conditions
cattle and other livestock and is then shed in their feces. Because at high temperatures for short periods of time destroys non-
of this, meats can easily become contaminated. Although the spore-forming pathogens and reduces the numbers of spoilage
initial contamination normally occurs on the surface, and the bac- organisms without significantly altering the flavor of food.
terial cells are easily destroyed by searing the exterior of meats pasteurization, p. 112
such as steaks, grinding the meat to create beef patties distributes ■ Cooking. Cooking, like pasteurization, can destroy non-
the pathogen throughout the product. Prevention then involves spore-forming organisms. Cooking obviously alters the char-
more thorough cooking so that enough heat reaches the center to acteristics of food, however. Heat distribution may be
kill all the E. coli cells, which is why hamburgers are a particularly uneven, resulting in survival of organisms in inadequately
troublesome source of infection. Outbreaks have also been linked heated regions.
to unpasteurized milk and various kinds of produce that were con-
■ Refrigeration. Refrigeration preserves food by slowing
taminated with animal manure.
the growth rate of microbes. Many organisms, including
MicroByte most pathogens, are unable to multiply at low temperatures.
21.7 million pounds of frozen ground beef patties were recalled in 2007 temperature requirements, p. 89
after they were linked to a multistate outbreak of E. coli O157:H7. ■ Freezing. Freezing stops microbial growth because water in
the form of ice is unavailable for biological reactions. Some
of the microbial cells will be killed by damage caused by ice
MicroAssessment 31.4
crystals, but those remaining can grow and spoil food once it
Foodborne intoxication results from consuming toxins produced is thawed.
by microbes growing in a food. Foodborne infection results from
■ Drying/reducing the aw. Drying foods or adding high
consumption of living organisms.
concentrations of sugars or salts inhibits microbial growth
9. How does boiling a home-canned food immediately prior to
by decreasing the available moisture. Eventually, however,
serving it prevent botulism?
molds may grow. drying food, p. 122
10. Which foodborne pathogen can cause hemolytic uremic syndrome?
■ Lowering the pH. Lowering the pH, either by adding acids
11. Why would a large number of competing microorganisms in a
food sample result in lack of sensitivity of culture methods for or encouraging fermentation by lactic acid bacteria, inhibits a
detecting pathogens? + wide range of spoilage organisms and pathogens.
■ Adding antimicrobial chemicals. Organic acids such as
propionic acid, benzoic acid, and sorbic acid are naturally
31.5 ■ Food Preservation occurring antimicrobial chemicals that are added to a vari-
ety of foods to inhibit fungal growth. Nitrates are added to
Learning Outcome cured meats to inhibit growth of Clostridium botulinum and
9. Describe the methods used to preserve foods. other organisms. Wine, fruit juices, and other products are
preserved by the addition of sulfur dioxide. chemical preserva-
Preventing the growth and metabolic activities of microorganisms tives, p. 121
that cause spoilage and foodborne illness preserves the quality of ■ Irradiation. Gamma radiation destroys microorganisms
food. Some methods of food preservation, such as drying and without significantly altering the flavor of foods such as
salting, have been known throughout the ages, whereas others spices and meats. irradiation, p. 115
have been discovered or developed more recently. The major
methods of preserving foods—high-temperature treatment, low-
temperature storage, addition of antimicrobial chemicals and
MicroAssessment 31.5
irradiation—are briefly summarized here and described in more
detail in chapter 5. Food spoilage can be eliminated or delayed by destroying
microorganisms or altering conditions to inhibit their growth.
■ Canning. The canning process destroys all spoilage and patho-
12. Why are the process temperatures for canning low-acid foods
genic organisms capable of growth at normal storage tempera- higher then ones for acidic foods?
tures. Low-acid foods are processed using steam under pressure
13. Why are nitrates added to cured meats?
(autoclaving) in order to reach temperatures high enough to
14. Microorganisms are often grouped according to their optimum
destroy the endospores of Clostridium botulinum. Acidic foods
growth temperatures. Which groups are most likely to spoil
do not require such high heat because C. botulinum cannot grow refrigerated foods? +
and produce toxin in those foods. canning, p. 113
760 Chapter 31 Food Microbiology

FUTURE CHALLENGES 31.1

Using Microorganisms to Nourish the World
To feed the world’s steadily increasing popu- Yeasts are considered the most promis- To a lesser extent, the use of bacteria as
lation, our limited natural resources must ing, large-scale source of single-cell pro- SCP is also being explored. The cyanobacte-
be used efficiently and new protein supplies tein. They multiply rapidly, are larger than rium Spirulina maxima can be cultivated in
must be developed. One potential solution bacteria, and are more readily acceptable alkaline lakes and then harvested and dried.
that addresses both these needs is to cultivate as a potential food. The most suitable type Unfortunately, this requires adequate sunlight
microorganisms as a protein source, using of yeast depends on the growth medium and warmth, making large-scale, year-round
industrial by-products currently considered employed, because different genera use dif- production impractical in most parts of the
wastes as the growth medium. fering carbohydrate sources and conditions world.
The term single-cell protein (SCP) was for growth. For example Kluyveromyces One of the chief concerns regarding the
coined in the 1960s to describe the use of uni- marxianus can be grown on whey, a by- consumption of microorganisms as a protein
cellular organisms such as yeasts and bacteria product of cheese-making; Saccharomyces source is their high concentration of nucleic
as a protein source. Today, the term generally cerevisiae can grow on molasses, a by- acid, primarily RNA. Yeast and bacteria contain
encompasses the use of multicellular microor- product of the sugar industry; and Candida as much as eight times more RNA per gram than
ganisms as well, and might more accurately utilis can multiply on cellulose-containing does meat. High levels of nucleic acid in the diet
be called microbial biomass. Although whole by-products of the pulp and paper industry. cause an increase in uric acid in the blood, which
cells can be consumed, a more palatable Most of these wastes must be supplemented can lead to gout and kidney stones. Chemical
alternative is to extract proteins from those with a nitrogen source, as well as various and enzymatic methods are being developed to
cells and use them to form textured protein vitamins and minerals, to support microbial decrease the nucleic acid in SCP without alter-
products. growth. ing the nutritional value of the protein.

Summary
31.1 ■ Factors Influencing the Growth Alcoholic Fermentations by Yeast (table 31.2)
of Microorganisms in Foods Wine is the product of the fermentation of sugars in fruit juices by spe-
Intrinsic Factors cially selected strains of Saccharomyces cerevisiae (figure  31.4). Beer
production is a multistep process designed to break down the starches
Bacteria require a high aw. Fungi often grow when the aw is too low
of grains such as barley to produce simple sugars, which can then serve
to support bacterial growth. Many bacterial species, including most
as a substrate for alcoholic fermentation by yeast (figure 31.5). Distilled
pathogens, are inhibited by acidic conditions. The nutritional content
spirits are produced using distillation to collect the alcohol generated
of a food determines the kinds of organisms that can grow in it. Rinds,
during fermentation. Vinegar is the product of the oxidation of alcohol
shells, and other coverings aid in protecting some foods from invasion
by the acetic acid bacteria. In bread-making, the CO2 produced by yeast
by microorganisms. Some foods contain natural antimicrobial chemi-
causes bread to rise; the alcohol is lost to evaporation (figure 31.6).
cals that help prevent spoilage.
Changes Due to Mold Growth (table 31.3)
Extrinsic Factors
Some cheeses and other foods are produced by encouraging the growth
Low temperatures halt or inhibit the growth of most foodborne micro-
of molds on foods. Soy sauce is made by allowing an Aspergillus
organisms. Psychrophiles and psychrotrophs, however, grow at refrig-
species to degrade a mixture of soybeans and wheat, which is then
eration temperatures. The presence or absence of O2 affects the type of
fermented in brine.
microbial population able to grow in a food.
31.3 ■ Food Spoilage
31.2 ■Microorganisms in Food Food spoilage is most often due to the metabolic activities of microor-
and Beverage Production ganisms as they grow and use the nutrients in the food.
Not only are fermented foods perceived as pleasant tasting, the Common Spoilage Bacteria
acids inhibit the growth of many spoilage organisms and foodborne Pseudomonas, Erwinia, Acetobacter, Alcaligenes, lactic acid bacteria,
pathogens. and bacteria that form endospores are important causes of food spoilage.
Lactic Acid Fermentations by the Lactic Acid Bacteria Common Spoilage Fungi
(table 31.1)
Fungi grow readily in acidic as well as low-moisture environments;
The tart taste of yogurt, pickles, sharp cheese, and some sausages is due therefore, fruits and breads are more likely to be spoiled by fungi than
to the metabolic products of the lactic acid bacteria. Starter cultures, by bacteria.
and sometimes rennin, are added to pasteurized milk to make cheese.
Other bacteria or fungi are sometimes added to cheese to give char- 31.4 ■ Foodborne Illness (table 31.4)
acteristic flavors or textures (figure  31.3). Pickling relies on naturally
occurring lactic acid bacteria. Commercial sausage production uses Foodborne Intoxication
starter cultures to rapidly decrease the pH and prevent the growth of Foodborne intoxication results from consuming a toxin produced by a
pathogens. microorganism growing in a food product (figure 31.7). Many strains of
 Part V Applied Microbiology 761

Staphylococcus aureus produce a toxin that, when ingested, causes nau- and Campylobacter species are commonly associated with poultry
sea and vomiting. Botulism is caused by ingestion of a neurotoxin pro- products. Some outbreaks of E. coli O157:H7 have been traced to
duced by the anaerobic, spore-forming, Gram-positive rod Clostridium undercooked contaminated hamburger patties and produce contami-
botulinum. As an added safety measure, low-acid home-canned foods nated with manure.
should be boiled at least 15 minutes immediately before serving it to
destroy any botulinum toxin that could be present. 31.5 ■ Food Preservation
Food spoilage can be eliminated or delayed by destroying microorgan-
Foodborne Infection isms or altering conditions to inhibit their growth. Methods used to
Foodborne infection requires the consumption of living organ- preserve foods include canning, pasteurization, cooking, refrigeration,
isms (figure  31.8). Thorough cooking of food immediately before freezing, reducing the aw, lowering the pH, adding antimicrobial chemi-
eating it will kill bacteria, thereby preventing infection. Salmonella cals, and irradiation.

Review Questions
Short Answer 8. Which group of organisms most commonly spoils breads, fruits,
1. What is the purpose of rennin in cheese-making? and dried foods?
2. What causes the bluish-green veins to form in blue cheese? a) Acetobacter b) Fungi
3. What causes the holes to form in Swiss cheese? c) Lactic acid bacteria d) Pseudomonas
4. What is the difference between traditional acidophilus milk and e) Saccharomyces
sweet acidophilus milk? 9. Which of the following organisms cause foodborne intoxication?
5. What is the purpose of the mashing step in beer-making? a) E. coli O157:H7 b) Campylobacter species
6. Explain how Alcaligenes species cause “ropiness” in raw milk. c) Lactobacillus species d) Salmonella species
e) Staphylococcus aureus
7. Explain the significance of Aspergillus flavus in grain products.
10. Canned pickles require less stringent heat processing than canned
8. Explain the typical sequence of events that lead to botulism.
beans, because pickles
9. Explain the typical sequence of events that lead to staphylococcal
a) contain fewer nutrients.
food poisoning.
b) are more acidic.
10. How does canning differ from pasteurization?
c) have a lower aw.
Multiple Choice d) contain antimicrobial chemicals.
e) are less likely to be contaminated with endospores.
1. The aw of a food product reflects which of the following?
a) Acidity of the food Applications
b) Presence of antimicrobial constituents such as lysozyme
1. A small cheese-manufacturing company in Wisconsin is looking
c) Amount of water available
for ways to reduce the costs of disposing of whey, a cheese by-
d) Storage atmosphere product. As a food microbiologist, what would you suggest that the
e) Nutrient content company do with the thousands of liters of whey being produced
2. Most spoilage bacteria cannot grow below an aw of per month so the company can actually profit from it?
a) 0.3. b) 0.5. c) 0.7. d) 0.9. e) 1.0. 2. A microbiologist is troubleshooting a batch of home-brewed ale
3. What is a generally minimum pH for growth and toxin production that did not ferment properly. She noticed that the alcohol content
by Clostridium botulinum and other foodborne pathogens? was only 2%, well below the desired level. Microscopic examina-
a) 8.5 b) 7.0 c) 6.5 d) 4.5 e) 2.0 tion showed numerous yeast cells. Chemical analysis indicated low
levels of sugar, high levels of CO2, and large amounts of protein
4. Benzoic acid is an antimicrobial chemical naturally found in which
in the liquid. What did the microbiologist conclude as the probable
of the following foods?
cause of the beer not coming out properly?
a) Apples b) Cranberries c) Eggs
d) Milk e) Yogurt Critical Thinking +
5. Which of the following is often added to wine to inhibit growth of 1. It has been argued that the nature of the growth of fungi in
the natural microbial population of grapes? Roquefort cheese, indicated by the appearance of bluish-green
a) Benzoic acid b) Lactic acid c) Carbon dioxide veins, is evidence that these fungi require O2 for growth. How does
d) Sulfur dioxide e) Oxygen this evidence lead to the conclusion?
6. In the brewing process, the sugar and nutrient extract obtained by 2. In the production of sauerkraut, a natural succession of lactic acid
soaking germinated grain in warm water is called bacteria is observed growing in the product. What causes the suc-
a) baker’s yeast. b) hops. c) malt. cession? What does this tell you about the optimal growth condi-
tions of the different species of lactic acid bacteria?
d) must. e) wort.
7. Which of the following genera is used in bread, wine, and beer
production?
a) Lactobacillus b) Pseudomonas c) Saccharomyces
d) Streptococcus e) Staphylococcus
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Appendix I
Microbial Mathematics

Because prokaryotes are very tiny and can multiply to very large The same prefix designations can be used for weights. The
numbers of cells in short time periods, convenient and simple ways basic unit of weight is the gram, abbreviated g. Approximately
are used to indicate their numbers without resorting to many zeros 450 grams are in a pound.
before or after the number. This is one reason why it is important
1 milligram = 1 mg = 0.001 gram = 10–3 g
to understand the metric system, which is used in scientific mea-
surements. 1 microgram = 1 μg = 0.000001 gram = 10–6 g
The basic unit of measure is the meter, which is equal to about 1 nanogram = 1 ng = 0.000000001 gram = 10–9 g
39 inches. All other units are fractions of a meter: 1 picogram = 1 pg = 0.000000000001 gram = 10–12 g

1 decimeter is one tenth = 0.1 meter Note that the number of zeros before the 1 is one less than the
1 centimeter is one hundredth = 0.01 meter exponent.
The value of the number is obtained by multiplying the base
1 millimeter is one thousandth = 0.001 meter
by itself the number of times indicated by the exponent.
Because prokaryotes are much smaller than a millimeter, Thus, 101 = 10 × 1 = 10
even smaller units of measure are used. A millionth of a meter is a
102 = 10 × 10 = 100
micrometer = 0.000001 meter, and is abbreviated μm. This is the
most frequently used size measurement in microbiology, because 103 = 10 × 10 × 10 = 1,000
bacteria are in this size range. For comparison, a human hair is When the exponent is negative, the base and exponent are
about 75 μm wide. divided into 1.
Because it is inconvenient to write so many zeros in front
of the 1, an easier way of indicating the same number is through For example, 10–2 = 1/10 × 1/10 = 1/100 = 0.01
the use of superscript, or exponential, numbers (exponents).
When multiplying numbers having exponents to the same
One hundred dollars can be written 102 dollars. The 10 is called
base, the exponents are added.
the base number and the 2 is the exponent. Conversely, one
hundredth of a dollar is 10–2 dollars; thus the exponent is nega- For example, 103 × 102 = 105 (not 106)
tive. The base most commonly used in biology is 10 (which is
designated as log10). The above information can be summarized When dividing numbers having exponents to the same base,
as follows: the exponents are subtracted.

1 millimeter = 1 mm = 0.001 meter = 10–3 meter For example, 105 ÷ 102 = 103
1 micrometer = 1 μm = 0.000001 meter = 10–6 meter In both cases, only if the bases are the same can the exponents
1 nanometer = 1 nm = 0.000000001 meter = 10–9 meter be added or subtracted.

A–1
 Appendix II
Pronunciation Key for Bacterial,
Fungal, Protozoan, and Viral Names
A Corynebacterium diphtheriae (koh-ryne-nee-bak-teer-ee-um dif-theer-
ee-ee)
Acetobacter (a-see'-toe-back-ter) Coxsackievirus (cock-sack-ee-vi-rus)
Achromobacter (a-krome-oh-back-ter) Cryptococcus neoformans (krip-toe-cock-us knee-oh-for-manz)
Acinetobacter (a-sin-et-oh-back-ter) Cytophaga (sigh-taw-fa-ga)
Actinomyces israelii (ak-tin-oh-my-seez iz-ray-lee-ee)
Actinomycetes (ak-tin-oh-my-seats)
Adenovirus (ad-eh-no-vi-rus) D
Agrobacterium tumefaciens (ag-rho-bak-teer-ee-um too-meh-faysh- Desulfovibrio (dee-sul-foh-vib-ree-oh)
ee-enz)
Alcaligenes (al-ka-li-jen-ease)
Amoeba (ah-mee-bah)
E
Arbovirus (are-bow-vi-rus) Eikenella corrodens (eye-keh-nell-ah kor-roh-denz)
Aspergillus niger (ass-per-jill-us nye-jer) Entamoeba histolytica (en-ta-mee-bah his-toh-lit-ik-ah)
Aspergillus oryzae (ass-per-jill-us or-eye-zee) Enterobacter (en-ter-oh-back-ter)
Azolla (aye-zol-lah) Enterococcus faecalis (en-ter-oh-kock-us fee-ka-liss)
Azotobacter (ay-zoh-toe-back-ter) Enterovirus (en-ter-oh-vi-rus)
Epidermophyton (eh-pee-der-moh-fy-ton)
Epulopiscium (ep-you-low-pis-se-um)
B Escherichia coli (esh-er-ee-she-ah koh-lee)
Bacillus anthracis (bah-sill-us an-thra-siss)
Bacillus cereus (bah-sill-us seer-ee-us) F
Bacillus coagulans (bah-sill-us coh-ag-you-lans)
Flavivirus (flay-vih-vi-rus)
Bacillus fastidiosus (bah-sill-us fas-tid-ee-oh-sus)
Flavobacterium (flay-vo-back-teer-ee-um)
Bacillus subtilis (bah-sill-us sut-ill-us)
Francisella tularensis (fran-siss-sell-ah tu-lah-ren-siss)
Bacillus thuringiensis (bah-sill-us thur-in-jee-en-sis)
Bacteroides (back-ter-oid-eez) Frankia (frank-ee-ah)
Baculovirus (back-you-low-vi-rus) Fusobacterium (fu-zoh-back-teer-ee-um)
Bdellovibrio (del-o-vib-re-oh)
Beggiatoa (beg-gee-ah-toe-ah) G
Beijerinckia (by-yer-ink-ee-ah) Gallionella (gal-ee-oh-nell-ah)
Bordetella pertussis (bor-deh-tell-ah per-tuss-iss) Gardnerella vaginalis (gard-nee-rel-lah va-jin-al-is)
Borrelia burgdorferi (bor-real-ee-ah berg-dor-fir-ee) Geobacillus stearothermophilus (gee-oh-bah-sill-us steer-oh-ther-
Bradyrhizobium (bray-dee-rye-zoe-bee-um) maw-fill-us)
Brucella abortus (bru-sell-ah ah-bore-tus) Giardia lamblia (jee-are-dee-ah lamb-lee-ah)
Gluconobacter (glue-kon-oh-back-ter)
C Gonyaulax (gon-ee-ow-lax)
Gymnodinium breve (jim-no-din-i-um brev-eh)
Campylobacter jejuni (kam-peh-low-back-ter je-june-ee)
Candida albicans (kan-did-ah al-bi-kanz)
Caulobacter (caw-loh-back-ter) H
Ceratocystis ulmi (see-rah-toe-sis-tis ul-mee) Haemophilus influenzae (hee-moff-ill-us in-flew-en-zee)
Chlamydia trachomatis (klah-mid-ee-ah trah-ko-ma-tiss) Helicobacter pylori (he-lih-koh-back-ter pie-lore-ee)
Chlamydophila pneumoniae (klah-mid-o-fil-ah new-moan-ee-ee) Hepadnavirus (hep-ad-nah-vi-rus)
Claviceps purpurea (kla-vi-seps purr-purr-ee-ah) Hepatitis virus (hep-ah-ti-tis vi-rus)
Clostridium acetobutylicum (kloss-trid-ee-um a-seat-tow-bu-till- Herpes simplex (her-peas sim-plex)
i-kum) Herpes zoster (her-peas zoh-ster)
Clostridium botulinum (kloss-trid-ee-um bot-you-line-um) Histoplasma capsulatum (his-toh-plaz-mah cap-su-lah-tum)
Clostridium difficile (kloss-trid-ee-um dif-fi-seal) Hyphomicrobium (high-foh-my-krow-bee-um)
Clostridium perfringens (kloss-trid-ee-um per-frin-gens)
Clostridium tetani (kloss-trid-ee-um tet-an-ee)
Coccidioides immitis (cock-sid-ee-oid-eez im-mi-tiss) I
Coronavirus (kor-oh-nah-vi-rus) Influenza virus (in-flew-en-za vi-rus)

A–2
 Appendix II Pronunciation Key for Bacterial, Fungal, Protozoan, and Viral Names A–3

K Propionibacterium shermanii (proh-pee-ah-nee-bak-teer-ee-um

sher-man-ee-ee)
Klebsiella pneumoniae (kleb-see-ell-ah new-moan-ee-ee) Proteus mirabilis (proh-tee-us mee-rab-il-us)
Pseudomonas aeruginosa (sue-dough-moan-ass aye-rue-gin-o-sa)
L
Lactobacillus brevis (lack-toe-ba-sil-lus bre-vis) R
Lactobacillus bulgaricus (lack-toe-ba-sil-lus bull-gair-i-kus) Rabies virus (ray-bees vi-rus)
Lactobacillus casei (lack-toe-ba-sil-us kay-see-ee) Retrovirus (re-trow-vi-rus)
Lactobacillus plantarum (lack-toe-ba-sil-us plan-tar-um) Rhabdovirus (rab-doh-vi-rus)
Lactobacillus thermophilus (lack-toe-ba-sil-us ther-mo-fil-us) Rhinovirus (rye-no-vi-rus)
Lactococcus lactis (lack-toe-kock-us lak-tiss) Rhizobium (rye-zoh-bee-um)
Legionella pneumophila (lee-jon-ell-ah new-moh-fill-ah) Rhizopus nigricans (rise-oh-pus nye-gri-kanz)
Leptospira interrogans (lep-toe-spire-ah in-ter-roh-ganz) Rhizopus stolon (rise-oh-pus stoh-lon)
Leuconostoc citrovorum (lew-kow-nos-tok sit-ro-vor-um) Rhodococcus (roh-doh-koh-kus)
Listeria monocytogenes (lis-tear-ee-ah mon-oh-sigh-to- jen-eze) Rickettsia rickettsii (rik-kett-see-ah rik-kett-see-ee)
Rotavirus (row-tah-vi-rus)
M Rubella virus (rue-bell-ah vi-rus)
Rubeola virus (rue-bee-oh-la vi-rus)
Malassezia (mal-as-seez-e-ah)
Methanobacterium (me-than-oh-bak-teer-ee-um)
Methanococcus (me-than-oh-ko-kus) S
Microsporum (my-kroh-spore-um) Saccharomyces carlsbergensis (sack-ah-row-my-sees karls-berg-
Mobiluncus (moh-bi-lun-kus) en-siss)
Moraxella catarrhalis (more-ax-ell-ah kah-tah-rah-liss) Saccharomyces cerevisiae (sack-ah-row-my-sees sara-vis-ee-ee)
Moraxella lacunata (more-ax-ell-ah lak-u-nah-tah) Salmonella enterica (sall-moh-nell-ah en-ter-i-kah)
Mucor (mu-kor) Serratia marcescens (ser-ray-sha mar-sess-sens)
Mycobacterium leprae (my-koh-bak-teer-ee-um lep-ree) Shigella dysenteriae (shig-ell-ah diss-en-tair-ee-ee)
Mycobacterium tuberculosis (my-koh-bak-teer-ee-um too-ber-kew-loh- Spirillum volutans (spy-rill-um vol-u-tanz)
siss) Sporothrix schenckii (spore-oh-thrix shenk-ee-ee)
Mycoplasma pneumoniae (my-koh-plaz-mah new-moan-ee-ee) Staphylococcus aureus (staff-ill-oh-kok-us aw-ree-us)
Staphylococcus epidermidis (staff-ill-oh-kok-us epi-der-mid-iss)
N Streptobacillus moniliformis (strep-tow-bah-sill-us mon-ill-i-form-is)
Streptococcus agalactiae (strep-toe-kock-us a-ga-lac-tee-ee)
Neisseria gonorrhoeae (nye-seer-ee-ah gahn-oh-ree-ee) Streptococcus cremoris (strep-toe-kock-us kre-more-iss)
Neisseria meningitidis (nye-seer-ee-ah men-in-jit-id-iss) Streptococcus mutans (strep-toe-kock-us mew-tanz)
Neurospora sitophila (new-rah-spor-ah sit-oh-phil-ah) Streptococcus pneumoniae (strep-toe-kock-us new-moan-ee-ee)
Streptococcus pyogenes (strep-toe-kock-us pie-ah-gen-ease)
O Streptococcus salivarius (strep-toe-kock-us sal-ih-vair-ee-us)
Streptococcus thermophilis (strep-toe-kock-us ther-moh-fill-us)
Orthomyxovirus (or-thoe-mix-oh-vi-rus)
Streptomyces griseus (strep-toe-my-seez gree-see-us)
Oscillatoria (os-sil-la-tor-ee-ah)

P T
Thiobacillus (thigh-oh-bah-sill-us)
Papillomavirus (pap-il-oh-ma-vi-rus)
Torulopsis (tore-you-lop-siss)
Parainfluenza virus (par-ah-in-flew-en-zah vi-rus)
Treponema pallidum (tre-poh-nee-mah pal-ih-dum)
Paramecium (pair-ah-mee-see-um)
Trichomonas vaginalis (trick-oh-moan-as vag-in-al-iss)
Paramyxovirus (par-ah-mix-oh-vi-rus)
Trichophyton (trick-oh-phye-ton)
Parvovirus (par-vo-vi-rus)
Trypanosoma brucei (tri-pan-oh-soh-mah bru-see-ee)
Pasteurella multocida (pass-ture-ell-ah mul-toe-sid-ah)
Pediococcus soyae (ped-ih-oh-ko-kus soy-ee)
Penicillium camemberti (pen-eh-sill-ee-um cam-em-bare-tee) V
Penicillium roqueforti (pen-eh-sill-ee-um rok-e-for-tee) Varicella-zoster virus (var-ih-sell-ah zoh-ster vi-rus)
Peptostreptococcus (pep-to-strep-to-ko-kus) Vibrio cholerae (vib-ree-oh kahl-er-ee)
Phytophythora infestans (fy-toe-fy-thor-ah in-fes-tanz) Vibrio fischeri (vib-ree-oh fish-er-i)
Picornavirus (pi-kor-na-vi-rus)
Plasmodium falciparum (plaz-moh-dee-um fall-sip-air-um)
Plasmodium malariae (plaz-moh-dee-um ma-lair-ee-ee) W
Plasmodium ovale (plaz-moh-dee-um oh-vah-lee) Wolbachia (wol-bach-ee-ah)
Plasmodium vivax (plaz-moh-dee-um vye-vax)
Pneumocystis jiroveci (new-mo-sis-tis yee-row-vet-ze)
Poliovirus (poe-lee-oh-vi-rus) Y
Polyomavirus (po-lee-oh-mah vi-rus) Yersinia enterocolitica (yer-sin-ee-ah en-ter-oh-koh-lih-tih-kah)
Propionibacterium acnes (proh-pee-ah-nee-bak-teer-ee-um ak-neez) Yersinia pestis (yer-sin-ee-ah pess-tiss)
 Appendix III
Metabolic Pathways

FIGURE III.1 The Entner-Doudoroff Pathway CH2O P

O
H H
Glucose 6- H
phosphate OH H
HO OH

H OH
+
NADP
Glucose 6-
phosphate
dehydrogenase
NADPH

CH2O P
O
H
6-phosphoglucono- H
O
d-lactone OH H
HO

H OH

H2O
Lactonase

COO-

H C OH

HO C H
6-phospho-
gluconate
H C OH

H C OH

CH2O P

6-phosphogluconate
dehydrase H2O

COO-

C O

CH2
2-keto-3-deoxy-6-
phosphogluconate H C OH

H C OH

CH2O P
KDPG aldolase
CHO COO-
Glyceraldehyde
3-phosphate H C OH C O

CH2O P CH3
Pyruvate

A–4
 Appendix III Metabolic Pathways A–5

Prokaryotes Eukaryotes
CH2OH CH2OH
O O
H H H H
H H
OH H OH H
HO OH HO OH

H OH H OH
Glucose Glucose
PEP ATP
Pyruvate Hexokinase
Group ADP
transport CH2O P
enzymes O
H H
Glucose 6-
OH H phosphate
HO OH

H OH
Phospho-
hexose
isomerase
O
P OH2C CH2OH
Fructose 6-
H HO
H OH phosphate

OH H
ATP Phosphofructo-
kinase
ADP

O
P OH2C CH2O P

OH Fructose 1,6-
H
H OH bisphosphate

OH H

Fructose bisphosphate
aldolase

CHO CH2O P
Dihydroxy-
Glyceraldehyde
H C OH C O acetone
3-phosphate
phosphate
CH2O P CH2OH

Pi

NAD
+ Glyceraldehyde-
3-phosphate
+ dehydrogenase
NADH + H

COO P ADP ATP COO- COO-

2-phospho-
H C OH H C OH H C O P
Phosphoglycerate Phosphoglycerate glycerate
CH2O P kinase CH2O P mutase CH2OH
1,3-bisphospho- 3-phospho-
glycerate glycerate
Enolase
H2O
COO- ATP ADP COO-

C O C O P
Pyruvate
CH3 kinase CH2
FIGURE III.2 The Embden-Meyerhof-Parnas Pathway Pyruvate
Commonly called glycolysis or the glycolytic pathway. Phosphoenolpyruvate
A–6 Appendix III Metabolic Pathways

COO-

CH2O P CH2O P H C OH
+
O NADP NADPH O H2O
H H H HO C H
H H
O
OH H OH H H C OH
HO OH Glucose 6- HO
phosphate H C OH
H OH dehydrogenase H OH
Glucose 6- 6-phosphoglucono- CH2O P
phosphate d-lactone 6-phospho-
gluconate
+
6-phosphogluconate NADP
dehydrogenase
NADPH
CO2
CH2OH
Phosphopentose
C O epimerase

H C OH

Phosphopentose H C OH Phosphopentose
isomerase epimerase
CH2O P
CHO CH2OH
Ribulose 5-
H C OH phosphate C O

H C OH HO C H

H C OH H C OH

CH2O P CH2O P
Phosphoglucoisomerase

Ribose 5- Xylulose 5-
phosphate Transketolase phosphate

CHO CH2OH

H C OH C O

CH2O P HO C H
Glyceraldehyde
H C OH
3-phosphate
H C OH

Transaldolase H C OH
CH2OH
CH2O P
C O Sedoheptulose
CH2OH
7-phosphate
HO C H
CHO C O
H C OH
H C OH HO C H
H C OH
H C OH H C OH
CH2O P
CH2O P CH2O P
Fructose 6- Erythrose 4- Xylulose 5-
phosphate phosphate phosphate
CH2OH
Transketolase
C O
CHO
HO C H
H C OH
H C OH
CH2O P
H C OH Glyceraldehyde
3-phosphate
CH2O P
Fructose 6-
FIGURE III.3 The Pentose Phosphate Pathway phosphate
 Appendix III Metabolic Pathways A–7

CH3 C S CoA
Acetyl CoA

O
Malate C COO-
dehydrogenase Citrate synthase
CH2 COO- CoA SH
+ +
Oxaloacetate H2O
NADH + H CH2 COO-
HO CH COO- NAD
+

HO C COO-
CH2 COO-
L-Malate CH2 COO-
Citrate

Aconitase
Fumarase Fe
2+

H2O H2O

CH2 COO-
H C COO-
-OOC C COO-
C H
Fumarate CH COO-
Cis-aconitate

FADH2

Succinate H2O
dehydrogenase
Aconitase
2+
FAD Fe

CH2 COO-

CH2 COO- CH2 COO-

Succinate
CH COO-
GTP
HO CH COO-
CoA SH Isocitrate

Succinyl- GDP + Pi
CoA +
NAD
synthase
CH2 COO-

CH2 NADH + H
+

+
Isocitrate
O C S CoA NADH + H dehydrogenase
Succinyl-CoA +
CO2
NAD
a-ketoglutarate CH2 COO-
dehydrogenase complex
CH2 2+
Mn
CoA SH
O C COO-
CO2 a-ketoglutarate

FIGURE III.4 The Tricarboxylic Acid Cycle (TCA Cycle)

Also referred to as the Krebs cycle or the citric acid cycle.
 Appendix IV
Answers to Multiple Choice Questions

Chapter 1 5. B Chapter 10 5. C Chapter 19 5. A Chapter 28

1. C 6. C 1. E 6. A 1. A 6. C
1. C
7. E 7. E 7. D
2. C 2. B 2. B 2. D
8. D 8. A 8. C
3. E 3. E 3. A 3. A
9. C 9. C 9. E
4. B 4. B 4. B
10. A 10. D 10. B 4. B
5. D 5. D 5. E
5. C
6. A Chapter 6 6. B Chapter 15 6. B Chapter 24
7. C 7. C 7. B 6. A
1. D 1. E 1. C 7. A
8. A 8. D 8. E
2. C 2. E 2. A
9. A 9. B 9. C 8. A
3. D 3. D 3. B
10. D 4. E 10. E 4. D 10. E 4. D 9. E
5. B 5. B 5. A 10. E
Chapter 2 Chapter 11 Chapter 20
6. A 6. D 6. D
1. C 1. E 1. D Chapter 29
7. D 7. D 7. C
2. A 2. B 2. C 1. A
8. D 8. A 8. D
3. E 3. E 3. A
9. D 9. B 9. C 2. A
4. A 4. B 4. B
10. E 10. D 10. E 3. C
5. B 5. D 5. A
6. C Chapter 7 6. A Chapter 16 6. B Chapter 25 4. D
7. B 7. D 7. D 5. A
1. B 1. C 1. D
8. D 2. A 8. B 2. A 8. D 2. D 6. D
9. E 3. A 9. C 3. D 9. A 3. C 7. C
10. A 4. A 10. D 4. D 10. C 4. B 8. C
Chapter 3 5. C Chapter 12 5. A Chapter 21 5. C 9. B
6. C 6. C 6. B 10. A
1. A 7. C 1. C 7. C 1. E 7. A
2. A 8. B 2. A 8. D 2. C 8. B Chapter 30
3. E 9. D 3. C 9. D 3. E 9. C 1. B
4. B 10. D 4. A 10. A 4. A 10. C
5. E 5. B 5. B 2. B
6. D Chapter 8 6. C Chapter 17 6. A Chapter 26 3. A
7. A 1. C 7. A 1. A 7. A 1. B 4. A
8. C 2. A 8. A 2. E 8. C 2. C 5. C
9. D 3. D 9. C 3. A 9. D 3. C 6. E
10. A 4. A 10. B 4. D 10. D 4. D 7. C
5. B 5. E 5. C 8. C
Chapter 4 Chapter 13 Chapter 22
6. D 6. C 6. E 9. B
1. D 7. A 1. C 7. A 1. E 7. A
2. A 2. A 2. E 10. E
8. B 8. C 8. A
3. B 9. A 3. C 9. A 3. A 9. B Chapter 31
4. B 10. B 4. D 10. A 4. E 10. E
5. B 5. E 5. E 1. C
6. E Chapter 9 6. C Chapter 18 6. A Chapter 27 2. D
7. C 1. C 7. D 1. C 7. C 1. E 3. D
8. A 2. C 8. A 2. D 8. A 2. A 4. B
9. C 3. E 9. A 3. D 9. D 3. C 5. D
10. B 4. B 10. D 4. E 10. D 4. E 6. E
5. B 5. D 5. C
Chapter 5 Chapter 14 Chapter 23 7. C
6. C 6. A 6. C
1. C 7. B 1. B 7. C 1. B 7. C 8. B
2. D 8. B 2. E 8. D 2. E 8. A 9. E
3. A 9. B 3. A 9. A 3. C 9. A 10. B
4. B 10. D 4. B 10. C 4. A 10. E
A–8
This page intentionally left blank
Glossary

abscess A localized collection of pus within a tissue. ADCC (antibody-dependent cellular cytotoxicity) alpha (α) hemolysis Type of hemolysis characterized
A-B toxin Exotoxin composed of an active subunit Killing of antibody-coated target cells by natural by a zone of greenish clearing around colonies grown
(A subunit) and a binding subunit (B subunit). killer cells, granulocytes, or macrophages. on blood agar.
accessory pigments Photosynthetic pigments such as adenosine diphosphate (ADP) The acceptor of alternative pathway Pathway of complement
carotenoids that capture light energy not absorbed by free energy in cells; that energy is used to add an activation initiated by the binding of a complement
chlorophylls. inorganic phosphate (Pi) to ADP, generating ATP. protein (C3b) to cell surfaces.
acid-fast An organism that retains the primary stain adenosine triphosphate (ATP) The energy currency alternative sigma factor A sigma factor that
in the acid-fast staining procedure. of cells. Hydrolysis of its unstable phosphate bonds recognizes promoters controlling genes needed only
acid-fast staining A procedure used to stain certain can be used to power endergonic (energy-consuming) in non-routine situations.
microorganisms, particularly Mycobacterium reactions. Ames test A test that screens for potential
species, that do not readily take up dyes. adhesin Component of a microorganism that is used carcinogens by measuring the ability of a substance to
acidophiles Organisms that grow optimally at a pH to bind to surfaces. increase the mutation frequency in a bacterial strain.
below 5.5. adhesion molecule Molecule on the surface of a cell amino acids Subunits of a protein molecule.
acquired resistance Development of antimicrobial that allows that cell to adhere to other cells. ammonification The decomposition process that
resistance through spontaneous mutation or adjuvant Substance that increases the immune converts organic nitrogen into ammonia (NH3).
acquisition of new genetic information. response to antigen. amphibolic pathways Metabolic pathways that play
actin filaments Cytoskeletal structures of eukaryotic ADP Abbreviation for adenosine diphosphate. roles in both catabolism and anabolism.
cells that allow movement within the cell. advanced treatment Any physical, chemical, or amylases Enzymes that digest starches.
actinomycetes Filamentous bacteria; many are biological purification process beyond secondary anabolism Cellular processes that synthesize and
valuable because they produce antibiotics. treatment of wastewater. assemble the subunits of macromolecules, using the
activated macrophages Macrophages stimulated by aerobic respiration Metabolic process in which energy of ATP; biosynthesis.
cytokines to enlarge and become metabolically active, electrons are transferred from the electron transport anaerobe container A specialized container that can
with greatly increased capability to kill and degrade chain to molecular oxygen (O2). maintain anaerobic conditions; a chemical reaction in
intracellular organisms and materials. aerosol Material dispersed into the air as a fine mist. a packet generates those conditions.
activated sludge method A method of sewage aerotaxis Movement toward or away from O2. anaerobic Without molecular oxygen (O2).
treatment in which wastes are degraded by complex aerotolerant anaerobe Organism that can grow in anaerobic chamber An enclosed compartment
populations of aerobic microorganisms. the presence of O2 but never uses it as a terminal maintained in an anaerobic environment; a special
activation energy Initial energy required to break a electron acceptor; an obligate fermenter. port is used to add or remove items.
chemical bond. affinity maturation The “fine-tuning” of the fit anaerobic digestion Process that uses anaerobic
activator In gene regulation, a protein that enhances the of an antibody molecule for an antigen; it is due to microbes to degrade the sludge obtained during
ability of RNA polymerase to initiate transcription. mutations that occur as activated B cells multiply. wastewater treatment.
activator-binding site Nucleotide sequence that aflatoxin Potent toxin made by Aspergillus flavus. anaerobic respiration Metabolic process in
precedes an ineffective promoter. agar Polysaccharide extracted from marine algae that which electrons are transferred from the electron
active immunity Protective immunity produced by is used to solidify microbiological media. transport chain to a terminal electron acceptor
an individual in response to an antigenic stimulus. agar plate A Petri dish that contains a solidified other than O2.
active site Site on an enzyme to which the substrate culture medium. analytical study In epidemiology, a study done to
binds; also known as the catalytic site. agarose Highly purified form of agar used in gel identify risk factors associated with developing a
active transport Energy-consuming process by electrophoresis. certain disease.
which cells move molecules across a membrane and agglutination reaction In immunological testing, anammox Anoxic ammonium oxidation.
against a concentration gradient. clumping together of cells or particles by antibody anaphylaxis (See systemic anaphylaxis.)
active tuberculosis disease (ATBD) A chronic molecules. anion Negatively charged ion.
illness caused by Mycobacterium tuberculosis and AIDS Acquired immunodeficiency syndrome. anneal To form a double-stranded duplex from two
characterized by slight fever, progressive weight loss, AIDS-related complex (ARC) A group of complementary strands of nucleic acid.
night sweating, and persistent cough, often producing symptoms—fever, fatigue, diarrhea, and weight anoxic Lacking O2.
blood-streaked sputum. loss—that mark the onset of AIDS. anoxygenic phototrophs Photosynthetic bacteria that
acute bacterial endocarditis Acute infection of the alga (plural: algae) A primitive photosynthetic use H2S or organic compounds rather than water as a
internal surfaces of the heart. eukaryotic organism. source of electrons for reducing power.
acute infection Illness characterized by signs and alkalophiles Organisms that grow optimally at a pH antagonistic In antimicrobial therapy, a combination
symptoms that develop quickly but last a relatively above 8.5. of antimicrobial medications in which the action of
short time. allele One form of a gene. one interferes with the action of the other.
acute inflammation Short-term inflammatory allergen Antigen that causes an allergy. antenna pigments Pigments of photosynthetic
response, marked by a prevalence of neutrophils. allergy Hypersensitivity, especially of the IgE- organisms that make up a complex that acts as a
acute retroviral syndrome (ARS) Stage of HIV mediated type. funnel, capturing light energy and transferring it to
disease following the incubation stage; often includes allograft Organ or tissue transplanted between reaction-center chlorophyll.
flulike symptoms. genetically nonidentical members of the same species. antibacterial drug Chemical used to treat bacterial
adaptive immunity Protection provided by host allosteric Refers to an enzyme or other protein that infections.
defenses that develop throughout life; involves B cells contains a site to which a small molecule can bind antibiogram Antibiotic susceptibility pattern; used to
and T cells. and change the protein’s activity. distinguish between different bacterial strains.

The Glossary contains definitions for the majority of bold terms in the text.

G–1
G–2 Glossary

antibiotic Chemical produced by certain molds and response to the slightest force between the tip and the beta-(β) lactam drugs Group of antimicrobial
bacteria that kills or inhibits the growth of other sample. medications that inhibit peptidoglycan synthesis and
microorganisms. ATP Abbreviation for adenosine triphosphate. have a shared chemical structure called a β-lactam
antibiotic-associated diarrhea Diarrhea that occurs ATP synthase Protein complex that harvests the ring.
as a complication of taking antimicrobial medications. energy of a proton motive force to synthesize ATP. beta-(β) lactamase Enzyme that breaks the β-lactam
antibody Immunoglobulin protein produced by the attaching and effacing (A/E) lesions Intestinal ring of a β-lactam drug, thereby inactivating the
body in response to a substance; it reacts specifically damage characterized by pedestals that form medication.
with that substance. under bacterial cells as a result of induced actin bile Yellow-colored fluid produced by the liver that
anticodon Sequence of three nucleotides in a tRNA rearrangement in the intestinal cell. aids in the absorption of nutrients from the intestine.
molecule that is complementary to a codon in mRNA. attack rate The percentage of individuals developing binary fission Asexual process of reproduction in
antigen Molecule that reacts specifically with an illness in a population exposed to an infectious agent. which one cell divides to form two daughter cells.
antibody or lymphocyte. attenuated vaccine Vaccine composed of a binomial system System of naming an organism
antigen-antibody complex Antibodies bound to weakened form of a pathogen. with two Latin words that indicate the genus and
antigen; also called an immune complex. autoantibodies Antibodies that bind to “self” species.
antigen presentation Process in which animal cells molecules. biochemical oxygen demand (BOD) Measure of the
display antigen on MHC molecules for T cells to autoclave Device that uses steam under pressure to amount of biologically degradable organic material
inspect. sterilize materials. in water.
antigen-presenting cells (APCs) Cells such as autoimmune disease Disease produced as a result of biodiversity Diversity in the number of species
B cells, macrophages, and dendritic cells that can an immune reaction against one’s own tissues. inhabiting an ecosystem and their evenness of
present exogenous antigen to helper T cells. autoradiography Method that uses film to detect a distribution.
antigenic Induces an immune response. radioactive molecule. biofilm Polymer-encased microbial community.
antigenic drift Slight changes in a viral surface autotroph Organism that uses CO2 as its main carbon bioinformatics Developing and using computer
antigen that render antibodies made against the source. technology to store, retrieve, and analyze nucleotide
previous version only partially protective. auxotroph A microorganism that requires an organic sequence data.
antigenic shift Major change in a viral surface growth factor. biological vector Organism that transmits a pathogen
antigen that render antibodies made against the aw Abbreviation for water activity. and within which the pathogen can multiply to high
previous version ineffective. axon The long thin extension of a nerve cell. numbers.
antigenic variation Process by which routine bioluminescence Biological production of light.
changes occur in a microbial surface antigen. bacillus (plural: bacilli) Cylindrical-shaped biomass Total weight of all organisms in any
anti-human IgG antibodies Antibodies that bind the bacterium; also referred to as a rod. particular environment.
constant region of human IgG molecules. bacteremia Bacterial cells circulating in the bioremediation Process that uses microorganisms to
antimicrobial drug Chemical used to treat microbial bloodstream. degrade harmful chemicals.
infections; also called an antimicrobial. Bacteria One of the two domains of prokaryotes; all biosphere The sum of all the regions of the earth
antiparallel Describes opposing orientations of the medically important prokaryotes are in the domain where life exists.
two strands of DNA in the double helix. Bacteria. biosynthesis Cellular processes that synthesize and
antiretrovirals (ARVs) Medications used to treat bactericidal Kills bacteria. assemble the subunits of macromolecules, using the
HIV infections. bacteriochlorophyll Type of chlorophyll used by energy of ATP.
antiseptic A disinfectant that is non-toxic enough to purple and green bacteria; absorbs wavelengths of biotechnology The use of microbiological and
be used on skin. light that penetrate to greater depths and are not used biochemical techniques to solve practical problems
antiserum A preparation of serum containing by other photosynthetic organisms. and produce valuable products.
protective antibodies. bacteriocins Proteins made by bacteria that kill bioterrorism The intentional use of microbes or their
antitoxin An antibody preparation that protects certain other bacteria. toxins to cause harm.
against a given toxin. bacteriophage A virus that infects bacteria; often biotype A group of strains that have a characteristic
antiviral drug A medication that interferes with the abbreviated to phage. biochemical pattern different from other strains; also
infection cycle of a virus. bacteriostatic Inhibits the growth of bacteria. called a biovar.
apoptosis Programmed cell death. baker’s yeast Selected strains of Saccharomyces biovar (See biotype.)
apicomplexans A group of protozoa that penetrate cerevisiae that are used to make yeast bread. blood agar Type of nutrient-rich agar medium that
host cells by means of a structure called an apical balanced pathogenicity Host-parasite relationship in contains red blood cells and can be used to detect
complex. which the parasite persists in the host while causing hemolysis.
aquaporins Pore-forming membrane proteins that minimal harm. blood-brain barrier Property of the central nervous
specifically allow water to pass through. base (See nucleobase.) system blood vessels that restricts passage of
arbovirus Arthropod-borne virus. One of a large base analog Compound that structurally resembles a infectious agents and certain molecules (such as
group of RNA viruses carried by insects and mites nucleobase closely enough to be incorporated into a medications) into the brain and spinal cord.
that act as biological vectors. nucleotide in place of the natural nucleobase. BOD Abbreviation for biochemical oxygen demand.
Archaea One of the two domains of prokaryotes; base-pairing The hydrogen bonding of adenine (A) boil Painful localized collection of pus within the skin
many archaea grow in extreme environments. to thymine (T) and cytosine (C) to guanine (G). and subcutaneous tissue; a furuncle.
arthropod Classification grouping of invertebrate base substitution A mutation in which the wrong bone marrow Soft material that fills bone cavities
animals that includes insects, ticks, lice, and mites. nucleotide has been incorporated. and contains hematopoietic stem cells.
Arthus reaction Hypersensitivity reaction caused by basement membrane Thin layer of fibrous material botulinum toxin Toxin produced by Clostridium
immune complexes and neutrophils. that underlies epithelial cells. botulinum that can cause a fatal paralysis.
artificially acquired immunity Immunity acquired basophil Leukocyte with large dark-staining granules brewer’s yeast Specially selected strains of Saccharomyces
through artificial means such as vaccination or that contain histamine and other inflammatory cerevisiae that are used in beer-making.
administration of immune globulin. mediators; receptors on its surface bind IgE. bright-field microscope Type of light microscope
aseptic technique Use of specific methods and sterile batch culture A culture system such as a tube, flask, that illuminates the field of view evenly.
materials to exclude contaminating microbes from an or agar plate in which nutrients are not replenished broad host range plasmid A plasmid that can
environment. and wastes are not removed. replicate in a wide variety of unrelated bacteria.
asexual Reproduction not preceded by the union of B-cell receptor (BCR) Membrane-bound derivative broad-spectrum antimicrobials Antimicrobials that
cells or exchange of DNA. of the antibody that a B cell is programmed to make; inhibit or kill a wide range of microorganisms, often
asthma Immediate respiratory allergy resulting from it allows the B cell to recognize a specific epitope. including both Gram-positive and Gram-negative
the release of mediators from mast cells in the lower B cells Lymphocytes programmed to produce bacteria.
airways. antibody molecules. Bt-toxin Protein crystal naturally produced by
astrobiology Study of life in the universe. B lymphocytes (See B cells.) Bacillus thuringiensis as it forms endospores; toxic to
atom The basic unit of all matter. beta-(β) hemolysis Type of hemolysis insect larvae that consume it.
atomic force microscope Type of scanning probe characterized by a clear zone around a colony bubo Enlarged, tender lymph node characteristic of
microscope that has a tip mounted so it can bend in grown on blood agar. plague and some sexually transmitted infections.
 Glossary G–3

bubonic plague Form of plague that typically catabolism Cellular processes that harvest the energy chemiosmotic theory The theory that a proton gradient
develops when Yersinia pestis is injected during the released during the breakdown of compounds such as is formed by the electron transport chain and then used to
bite of an infected flea. glucose, using it to synthesize ATP. power ATP synthesis.
budding (1) Asexual reproductive technique that catabolite Product of catabolism. chemoautotrophs Organisms that use chemicals as
involves a pushing out of a part of the parent cell that catalase Enzyme that breaks down hydrogen peroxide a source of energy and CO2 as the major source of
eventually gives rise to a new daughter cell. (2) A (H2O2) to produce water (H2O) and oxygen gas (O2). carbon.
process by which some viruses are released from host catalyst Substance that speeds up the rate of a chemoheterotrophs Organisms that use chemicals
cells. chemical reaction without being altered or depleted in as a source of energy, and organic compounds as a
buffer Substance in a solution that acts to prevent the process. source of carbon.
changes in pH. catheter A flexible plastic or rubber tube inserted chemokine Cytokine important in chemotaxis of cells
bulking Overgrowth of filamentous microorganisms into the bladder or other body space, in order to drain of the immune system.
in sewage at treatment facilities; interferes with the it or deliver medication. chemolithoautotrophs Organisms that obtain
separation of the solid sludge from the liquid effluent. cations Positively charged ions. energy by oxidizing reduced inorganic compounds
burst size Number of newly formed virus particles CD markers Abbreviation for cluster of such as hydrogen gas (H2), and use CO2 as a source
released when the infected host cell lyses differentiation markers. of carbon.
CD4 lymphocytes T lymphocytes bearing the CD4 chemolithotrophs Organisms that obtain energy
Calvin cycle Metabolic pathway used by many markers; helper T cells are CD4 cells. by oxidizing reduced inorganic chemicals such as
autotrophs to incorporate CO2 into an organic form; CD8 lymphocytes T lymphocytes bearing the CD8 hydrogen gas (H2); in general, chemolithotrophs are
also called the Calvin-Benson cycle. markers; cytotoxic T cells are CD8 cells. chemolithoautotrophs.
cAMP Abbreviation for cyclic AMP. cDNA DNA obtained by using reverse transcriptase to chemoorganoheterotrophs Organisms that obtain
cancer Abnormally growing cells that can spread synthesize DNA from an RNA template in vitro; lacks both energy and carbon from organic compounds.
from their site of origin; malignant tumors. introns that characterize eukaryotic DNA. chemoorganotrophs Organisms that obtain
candidiasis Fungal diseases caused by Candida cell culture (or tissue culture) Cultivation of animal energy by oxidizing organic compounds such
albicans. or plant cells in the laboratory. as glucose; in general, chemoorganotrophs are
CAP Abbreviation for catabolite-activating protein, an cell envelope The layers surrounding the contents chemoorganoheterotrophs.
activator involved in carbon catabolite repression. of the cell; includes the cytoplasmic membrane, cell chemostat Device used to grow bacteria in the
capnophiles Organisms that require increased wall, and capsule (if present). laboratory that allows nutrients to be added and
concentrations of CO2 (5% to 10%) and cell wall Strong barrier that surrounds a cell, keeping waste products to be removed continuously.
approximately 15% O2. the contents from bursting out; in prokaryotes, chemotaxis Directed movement of an organism in
capped In eukaryotic gene expression, adding a peptidoglycan provides strength to the cell wall. response to a certain chemical in the environment.
methylated guanine derivative to the 5 end of the cell-mediated immunity (CMI) Immunity due a chemotherapeutic agent Chemical used as a
mRNA. T-cell response; also called cellular immunity. therapeutic medication to treat a disease.
capsid Protein coat that surrounds the nucleic acid of cellular immunity (See cell-mediated immunity.) chemotrophs Organisms that obtain energy by
a virus. cellulose Polymer of glucose subunits; principal oxidizing chemical compounds.
capsule A distinct thick gelatinous material that structural component of plant cell walls. chickenpox Disease characterized by widespread
surrounds some types of microorganisms; sometimes Centers for Disease Control and Prevention (CDC) itchy fluid-filled blisters; caused by varicella-zoster
required for an organism to cause disease. The U.S. government agency charged with the task of virus (VZV), a herpesvirus.
capsule stain A staining method used to observe controlling and preventing diseases and injuries. chlorophylls The primary light-absorbing pigments
capsules. central metabolic pathways Glycolysis, the TCA used in photosynthesis.
carbapenems Group of antimicrobial medications cycle, and the pentose phosphate pathway; the chloroplasts Organelles in photosynthetic eukaryotic
that interferes with peptidoglycan synthesis; very transition step is often considered a part of the TCA cells that harvest the energy of sunlight and use it
resistant to inactivation by β -lactamases. cycle. to synthesize ATP, which is then used to fuel the
carbapenem-resistant Enterobacteriaceae (CRE) central nervous system (CNS) Brain and spinal synthesis of organic compounds.
Members of the Enterobacteriaceae that are cord. chocolate agar Type of agar medium that contains
resistant to carbapenems, as well as nearly all other cephalosporins Group of antimicrobial medications lysed red blood cells; used to culture fastidious
antimicrobial drugs. that interfere with peptidoglycan synthesis. bacteria.
carbohydrate Compounds containing principally carbon, cerebrospinal fluid (CSF) Fluid produced within cholesterol Sterol found in animal cell membranes;
hydrogen, and oxygen atoms in a ratio of 1:2:1. the brain that surrounds the central nervous provides rigidity to eukaryotic membranes.
carbon catabolite repression A regulatory system. chromosome Structure that carries an organism’s
mechanism that allows cells to prioritize their use of cestode Tapeworm. hereditary information.
carbon/energy sources. chain terminator A dideoxynucleotide; when this chronic infections Infections that develop slowly and
carbon fixation Process of converting inorganic molecule is incorporated into a growing strand of persist for months or years.
carbon (CO2) to an organic form. DNA, no additional nucleotides can be added, so chronic inflammation Long-term inflammatory
carbuncle Painful infection of the skin and elongation of the strand stops. response, marked by the prevalence of macrophages,
subcutaneous tissues; manifests as a cluster of boils. challenge In immunology, to give an antigen to giant cells, and granulomas.
carcinogen A chemical or radiation that causes cancer. provoke an immunologic response in a subject chronic periodontitis A destructive inflammatory
cariogenic Causing dental caries, tooth decay. previously sensitized to the antigen. response that progressively damages the structures
carotenoids Type of accessory pigment that chancre Sore resulting from an ulcerating infection; that support the teeth.
increases the efficiency of light capture by the “hard chancre” of primary syphilis is typically cilium (plural: cilia) Short, projecting hairlike
absorbing wavelengths of light not absorbed by firm and painless. structure of locomotion in some eukaryotic cells,
chlorophylls. channel Pore-forming membrane protein that allows similar in function to a flagellum.
carrier (1) Type of protein found in cell membranes specific ions to diffuse into and out of a cell. cirrhosis Scarring of the liver that interferes with
that transports certain compounds across the chaperone Protein that helps other proteins fold normal liver function.
membrane; may also be called a permease or properly. clade Subtype of a virus such as HIV, distinguished
transporter protein. (2) A human or other animal that chemical bond Force that holds atoms together to by similar amino acid sequences of envelope proteins.
harbors a pathogen without noticeable ill effects. form molecules. class In classification, a collection of similar orders; a
case-control study In epidemiology, a study that chemically defined media A culture medium collection of several classes makes up a phylum.
compares activities of people who developed a composed of exact quantities of pure chemicals; class switching The process that allows a B cell
disease with those of people who did not. generally used for specific experiments when to change the antibody class it is programmed to
case-fatality rate Percentage of people dying of a nutrients must be precisely controlled. make.
specific disease within a specific time period. chemiosmotic gradient Accumulation of protons classical pathway Mechanism by which immune
caseous necrosis Type of localized tissue death immediately outside the cell or mitochondrial complexes activate the complement system.
resulting in a cheeselike consistency, characteristic matrix due to ejection of protons by the electron classification The arrangement of organisms into
of tuberculosis and certain other chronic infectious transport chain; also called the proton motive similar or related groups, primarily to provide easy
diseases. force. identification and study.
G–4 Glossary

clinical latency In HIV disease, the time period community All of the living organisms in a given contrast In microscopy, the number of different
during which the virus actively replicates without area. visible shades in a specimen.
causing obvious signs or symptoms of disease. competent In horizontal gene transfer, physiological convalescence Period of recuperation and recovery
clonal selection Process in which a lymphocyte’s condition in which a bacterial cell is capable of taking from an illness.
antigen receptor binds to an antigen, potentially up DNA. convergent evolution Process of evolution when two
allowing the lymphocyte to proliferate and then competitive inhibition Type of enzyme inhibition genetically different organisms independently develop
differentiate into an effector cell. that occurs when the inhibitor competes with the similar structures.
clone Group of cells derived from a single cell. normal substrate for binding to the active site. core genome DNA sequences found in all strains of a
closed system A system such as a tube, flask, or complement system Series of serum proteins particular species.
agar plate in which nutrients are not replenished involved with innate immunity; they can be rapidly corepressor Molecule that binds to an inactive
and wastes are not removed as microorganisms activated, contributing to protective outcomes repressor, thereby allowing it to function as a
grow. including inflammation, lysis of foreign cells, and repressor.
clumping factor Fibrinogen-binding virulence factor opsonization. coryneforms Gram-positive cells that are club-
of Staphylococcus aureus that serves as an identifying complementary In DNA structure, the nucleobases shaped and arranged to form V shapes and palisades;
characteristic. that characteristically hydrogen bond to one another; resembles the typical microscopic morphology of
clusters of differentiation (CD) markers Surface A (adenine) is complementary to T (thymine), and G Corynebacterium species.
molecules that allow scientists to distinguish subsets (guanine) is complementary to C (cytosine). co-stimulatory molecules Surface proteins expressed
of T cells and other white blood cells. complex medium Medium for growing bacteria by antigen-presenting cells (APCs) when the cell
CMI Abbreviation for cell-mediated immunity. that has some ingredients of variable chemical senses molecules that signify an invading microbe or
CO2 fixation Process of converting inorganic carbon composition. tissue damage; they help dendritic cells activate naive
(CO2) to an organic form. composite transposon A transposon composed of T cells.
coagulase Plasma-clotting virulence factor of one or more genes flanked by insertion sequences. Coulter counter An instrument that counts cells in a
Staphylococcus aureus that serves as an identifying composting The natural decomposition of organic suspension as they pass through a narrow channel.
characteristic. solid material. counterstain In a differential staining procedure, the
coccobacillus Rod that is so short it can be mistaken compound lipid Molecules that contain fatty acid and stain applied to impart a contrasting color to bacteria
for a coccus. glycerol in addition to an element other than carbon, that do not retain the primary stain.
coccus (plural: cocci) Spherical-shaped bacterial cell. hydrogen, or oxygen. covalent bond Strong chemical bond formed by the
codon Set of three adjacent nucleotides that encode compound microscope Microscope that uses sharing of electrons between atoms.
either an amino acid or the termination of the multiple magnifying lenses, thereby visually CRISPR system Mechanism by which bacterial cells
polypeptide. enlarging an object by a factor equal to the product of maintain a historical record of phage infections, and
coenzyme Non-protein organic compound that assists the magnification of each lens. thereby become immune to subsequent infections
some enzymes, acting as a loosely bound carrier of condenser lens Lens used to focus the illumination of by the same phages; the system also protects against
small molecules or electrons. a microscope; it does not affect the magnification. other types of foreign DNA.
cofactor Non-protein component required for th