Aerobic gram positive bacilli

and their pathogenetic

properties
Learning objectives

 Know the pathogenic aerobic

Gram positive bacilli
 Define virulence factors of agents
 Know the principal clinical
features and pathology
 the basis of diagnosis
Aerobic gram positive bacilli

“big fat rods” Bacillus

Small rods Corynebacterium

Listeria
 Aerobic and facultatively anaerobic
 Spore forming gram-positive bacilli
 ~50 Bacillus species

( Bergey’s Manual of Systematic

Bacteriology)

 Bacillus anthracis and Bacillus cereus

are significant human pathogens
Bacillus species and their diseases

Organism Diseases
Bacillus anthracis Anthrax
Bacillus cereus Gastroenteritis
emetic form
diarrheal form
Panophtalmitis
Opportunistic infections
Other Bacillus species Opportunistic infections
 Your patient is a 30-year-old man with a 2-cm lesion
on his arm. It began as a painless papule that
enlarged and, within a few days, ulcerated and
formed a black crust (eschar). He works in an
abattoir where his job is removing the hide from the
cattle. A Gram stain of fluid from the lesion reveals
large gram-positive rods. Which one of the following
bacteria is likely to be the cause?
(A) Bacillus anthracis
(B) Clostridium botulinum
(C) Clostridium perfringens
(D) Clostridium tetani
(E) Listeria monocytogenes
anthrax
 derives its name from the Greek word for coal,

anthracis,

 cause black, coal-like cutaneous eschars

 zoonotic disease

 a part of biological weapons program by

several countries
Bacillus anthracis
 1 by 3 mm
 usually is straight
 may be slightly curved
 ends of the bacilli are
truncated, not rounded
 tend to form into long
chains
 may appear similar to
streptobacilli
Bacillus anthracis
 catalase-positive
 nonmotile
 grow optimally in an
atmosphere of
enhanced carbon
dioxide
 Bacillus anthracis can switch
back and forth between two
states:

 the active “vegetative” form

 the dormant “spore” form

 form oval central spores
 nonswollen sporangium
 Spores are destroyed by
steam sterilization or
burning
 Resistant to detergents
mercury chloride-1/1000, 72h
potassium permanganate
4/100, 30 min Gram stain.
Hydrogen peroxide 3/100, 1h The spores are highly refractile to
light and resistant to staining.
Bacillus anthracis;

 virulent strains form a

single antigenic type of
capsule
poly-D-glutamate
polypeptide
B. anthracis capsule
 R variants are relatively avirulent
 Capsule production depends on a
plasmid, pX02;
 The poly-D-glutamyl capsule is itself
nontoxic,
 functions to protect the organism
against phagocytic engulfment
Anthrax Toxin
 a diffusible exotoxin plays a major role
in the pathogenesis of anthrax
 production of the anthrax toxin is
mediated by a temperature-sensitive
plasmid, pX01
 The toxin consists of three distinct
antigenic components.
Components of Exotoxin
Factor I: edema factor (EF)
 a calmodulin-dependent adenylate cyclase,
causes the elevation of intracellular cAMP
 responsible for the severe edema usually seen
in anthrax
Factor II: protective antigen (PA),
Factor III: known as the lethal factor (LF) t i vity
gical ac
b i ol o
i g ni f i cant
i b i t s nos
ac t o r exh
each f
Epidemiology 1
 It is carried by wild and domestic animals in
Asia, Africa and parts of Europe
 Most infections are reported in Iran, Turkey,
Pakistan and Sudan
 Anthrax primarily is a disease of herbivores
(cattle, sheep, goats, horses).
 Humans may be accidentally infected:
Epidemiology 2
anthrax is not a contagious disease
 That means it can’t be passed from one
person to another
 The only way to be infected is to come into
direct contact with anthrax spores
acquired by one of three routes:
inoculation, inhalation or ingestion
Cutaneous anthrax
(most common form)
 1-7 days after skin exposure and penetration of
B anthracis spores
 most commonly affects the upper extremities
 to a lesser extent, the head and neck.
 Hematogenous dissemination in 5-10% of
untreated cases.
 Your patient is a 30-year-old man with a 2-cm lesion
on his arm. It began as a painless papule that
enlarged and, within a few days, ulcerated and
formed a black crust (eschar). He works in an
abattoir where his job is removing the hide from the
cattle. A Gram stain of fluid from the lesion reveals
large gram-positive rods. Which one of the following
bacteria is likely to be the cause?
(A) Bacillus anthracis
(B) Clostridium botulinum
(C) Clostridium perfringens
(D) Clostridium tetani
(E) Listeria monocytogenes
Intestinal anthrax
 Ingesting B anthracis spores
 in 2-5 days severe abdominal pain,
hematemesis, bloody diarrhea, and fever.
 multiple ulcerative lesions throughout the GI
tract
 secondary to hematogenous spread
 difficult to recognise, and shock and death
may occur 2-5 days after onset.
Inhalational anthrax 1
Inhalational anthrax usually is fatal
 begins abruptly,

 in 1-3 days

 initially nonspecific symptoms,

a low-grade fever

a nonproductive cough
Inhalational anthrax 2

 substernal discomfort early in the illness

 Patients may improve temporarily

 then progresses rapidly with high fever,

 severe shortness of breath, tachypnea,

 cyanosis, hematemesis, and chest pain,

(mimic an acute myocardial infarction)

Lab Studies 1
 for cutaneous anthrax,

staining the ulcer

exudate with methylene

blue or Giemsa stain.

 B anthracis readily

grows on blood agar,

Lab Studies 2
suggesting extracutaneous
spread,
 blood culture.
 Serological diagnosis

specific antibodies
 PCR
TREATMENT
Penicillin, Ciprofloxacin
• IMMUNIZATION
Animals
> Live spore vaccine (Sterne strain)
Workers at Risk of Exposure
>Anthrax Vaccine Absorbed (AVA)
“Alum precipitated toxoid”
Corynebacterium-1

Club-shaped, Gram-positive rods

Arranged in V forms or Palisades

(Chinese letters)
Corynebacterium-2
 Facultative and NON spore
forming

• Nonmotile Rods
• Catalase +ve

 Most are relatively non fastidious

but give small colonies
Corynebacterium-3
 Most are non pathogenic and normal
flora on skin & mucous membranes
 Some are pathogens (rare) and
 Some are opportunistic (rare).
 Most encounters with
Corynebacterium spp. in the
laboratory are with “diphtheroids”
Diphtheria
 Toxin-producing strains of Corynebacterium
diphtheriae
• Pleomorphic Bacilli
• Cells Contain Metachromatic Granules.
 Transmission
 Direct person- to-person transmission by
intimate respiratory and physical contact.
 Cutaneous lesions are important in
transmission.

 Risk Groups
 In the pre-vaccine era, children were at
highest risk for respiratory diphtheria.
 Recently, diphtheria has primarily affected
adults in the sporadic cases reported
Incidence

1200 80

70
1000

ölüm sayısı
vaka sayısı

vaka 60
800
50
ölüm
600 40

30
400
20
200
10

0 0

yıllar
 PATHOGENICITY

corynebacteriophage ß,
which carries tox
 PATHOGENICITY
 Organism does not produce a systemic infection
 Diphtheria is a Toxaemia
 Exotoxin: Molecular wt 62000
 Toxin consisits 2 types of polypeptide
 One binds to host cells;
 the other then becomes internalized and inhibits
protein synthesis
 Has Special Affinity for Certain Tissues
"Mycocardium, Adrenals, Nerve Endings"
 causes Pseudomebrane
Complications
 Myocarditis, polyneuritis, and airway
obstruction are common complications
of respiratory diphtheria
 death occurs in 5%-10% of respiratory
cases
 Complications and deaths are much less
frequent in cutaneous diphtheria
 LABORATORY DIAGNOSIS

Specific Treatment Should beStarted

Immediately on Suspicion of Diphtheria

Specimen: Two Swabs from the Lesion

 Preparing slide
 Culture:

Loeffler's medium
Tellurite Blood Agar
Corynebacterium diphtheriae forms black
colonies on tellurit agar (left),
on blood agar colonies appear white (right)
Production of exotoxin can be determined by
in vivo or in vitro tests

• In-vivo Tests:
i. Subcutaneous Test:

ii. Intracutaneuos Test:

• In-vitro Test:
 Elek Test ( Gel Precipitaion Test)
 EIA
 PCR

• Tissue Culture:
 TREATMENT
> Moderate Cases:
i. Antitoxin:
ii. Antibiotic:
> Severe Cases:
i. Active Immunization,DPT
(diphtheria, pertussis, tetanus) vaccine
ii. Passive Immunization:
iii. Combined:
PREVENTION
Other Corynebacterium spp.

Corynebacterium jeikeium (JK)

- Opportunistic and Ab resistant
- Grown from sterile sites
Corynebacterium urealyticum
-Has a particular specialty of
alkaline encrusted cystitis
Each of the following statements concerning
Corynebacterium diphtheriae iscorrect EXCEPT:

(A) C. diphtheriae is a gram-positive rod that does

not form spores.
(B) Toxin production is dependent on the
organism’s being lysogenized by a
bacteriophage.
(C) Diphtheria toxoid should not be given to
children younger than 3 years because the
incidence of complications is too high.
(D) Antitoxin should be used to treat patients with
diphtheria
 Your patient in the pediatric intensive care unit is a 2-
week-old boy with a high fever and the signs of
meningitis. Gram stain of the spinal fluid reveals
small gram-positive rods. Colonies on blood agar
show a narrow zone of β-hemolysis.
 Which one of the following is the most likely cause of
his neonatal meningitis?

(A) Bacillus anthracis

(B) Bacillus cereus
(C) Clostridium perfringens
(D) Corynebacterium diphtheriae
(E) Listeria monocytogenes
Listeria monocytogenes
 Aerobic to Microaerophilic
 G+ve Coccobacilli, grows in Short Chains
 Has the listerial LPS
molecule chemically and biologically similar
to the classical lipopolysaccharide
 beta hemolytic colonies on blood agar plates
 Motile at 25C, BUT NOT at 37C
 a facultative intracellular
Listeria monocytogenes
Virulence Properties
 β-hemolysin (listeriolysin O),
lecithinase and phospholipase
 produced by virulent strain only
 oxygen labile
 antigenic
 disrupts phagocytic vacuole
 Virulence Properties
 Intracellular Growth
 bacterium escapes from host vacuole
 undergoes rapid division
 becomes encapsulated by short actin
filaments
 facilitates movement to periphery with
protrusion and then penetration to
neighboring cells
 never exposed to humoral immunity
 Listeriosis
two forms:
Food
Mammals products
Birds
Direct  neonatal
Fish
contact
Insect disease
Environment  adult disease.
Adult Disease
Normal adult are immunosuppressed
Resistant individuals
 Infection results in self- HIV infected and other
resolving flu-like immunosuppressed
symptoms patients with cancer
 and/or mild and in renal transplant
gastrointestinal recipients
disturbance  Gastroenteritis
 Chills and fever are due  Septicemia
to bacteremia  Meningitis
Neonatal disease
Mother usually has no symptomatic
illness

two forms:
 early onset disease, acquired
transplacetally in utero
 late onset disease acquired at birth
or soon after birth
 Neonatal Disease
 Granulomatosis infantiseptica - infection of
fetus in utero
 can lead to abortion, disease at birth (sepsis,
pneumonia, fetal distress, seizures, rash,
abscesses, and granulomas
 High mortality rate
 Exposure on vaginal delivery
 can result in meningitis or meningo-
encephalitis with sepsis within 2-3 weeks
LABORATORY DIAGNOSIS

Specimen: Depend on Syndrome:

Blood, CSF, Genital Tract Secretions

 monocytosis in the peripheral blood

 Gram-positive rods in a Gram stain
of smears of the cerebrospinal fluid
 Culture
 Control
Hygienic food processing and storage
Avoid uncooked food

Treatment
Penicillin or ampicillin alone or in
combination with gentamicin