Shree Swaminarayan College of Pharmacy.

2nd year Pharm.D

Pharmaceutical Microbiology

CH: 2
By: Anjali Luhana
 Different methods of classification of
microbes.
• Classification allows the orderly grouping of micro-organisms.
• At present no standard classification of bacteria is universally
accepted and applied, although Bergey's Manual of
Determinative Bacteriology is widely used as an authoritative
source.
• Methods of classification:
1) Adansonian or numerical classification: In most systems of
bacterial classification, the major groups are distinguished by
fundamental characters such as cell shape, Gram-stain reaction
and spore formation; genera and species are usually
distinguished by properties such as fermentation reactions,
nutritional requirements and pathogenicity.
• 2) DNA composition: The hydrogen bonding between guanine and
cytosine (G–C) base pairs in DNA is stronger than that between
adenine and thymine (A–T). Thus, the melting or denaturation
temperature of DNA (at which the two strands separate) is determined
primarily by the G + C content. At the melting temperature, the
separation of the strands brings about a marked change in the light
absorption characteristics at a wavelength of 260 nm, and this is
readily detected by spectrophotometry.
• 3) DNA homology: Another approach to classification is to arrange
individual organisms into groups on the basis of the homology of their
DNA base sequences. This exploits the fact that double strands re-form
(anneal) from separated strands during controlled cooling of a heated
preparation of DNA. This process can be readily demonstrated with
suitably heated homologous DNA extracted from a single species
• 4) Ribosomal RNA sequencing:
• The structure of ribosomal RNA (rRNA) appears to have been
highly conserved during the course of evolution, and close
similarities in nucleotide sequences reflect phylogenetic
relationships. Advances in technology have made nucleotide
sequencing relatively simple, and the rDNA sequences (and
other genes) of most medically important bacterial species are
available from a number of internet sites.
 Bacteria
• Bacteria are unicellular ,free –living ,microscopic microorganisms
capable of performing all the essential functions of life.
• Bacteria are prokaryotic microorganisms that do not contain
chlorophyll.

• Size of Bacteria:
• Bacteria are very small microorganisms which are visible under
the light microscope.
• Generally,cocci are about 1µm in diameter and bacilli are 1to 8µm
in length and 0.1 to 0.5 µm in width.
• Bacteria are stained by staining reagents and then visualized under
power of magnifications of compound microscope.
 Shape of bacteria

• On the basis of shape bacteria are classified as follows:

• 1`) Cocci: Cocci are small, spherical or oval cells.e.g.
Micrococcus.
• 2) Bacilli: It is derived from the Greek word ‘Bacillum’
meaning ‘stick’.
• There are rod shaped cells.e.g. Bacillus anthracis.
• In some of the bacilli the length of the cells may be equal to
width. such bacillary forms are known as coccobacilli.e.g.
Bracella.
• 3) Vibrios: They are comma shaped ,curved rods e.g. Vibrio
comma.
• 4) Spirilla : These are longer rigid rods with several curves or
coils. they have helical shape and rapid bodies e.g. Spirillum
ruprem.
• 5) Spirochetes : They are slender and flexuous spiral forms.
• 6) Actinomycetes: They are branching filamentous bacteria.
The characteristic shape is due to presence of a rigid cell wall
e.g. Streptomyces species.
• 7) Mycoplasmas: They are cell wall deficient bacteria and
hence do not possess a stable morphology. they occur as round
or oval bodies with interlacing filaments.
 Arrangement of bacterial cells
• The most common method of reproduction among bacteria is asexual
binary fission. In this process, each cell splits by forming two new
cells.
• Cocci appear in several characteristic arrangements or grouping.
• 1) Diplococci: Cocci that split along one plane only, tend to arrange
themselves in pairs.e.g. Diplococcus pneumoniae.
• 2) Streptococci: These cells divide in one plane and remain
attached ,to form chains.e.g. Streptococcus lactis.
• 3) Tetracocci : Tetracocci divide in two planes and live in groups of
four.e.g. Gaffyka tetragena.
• 4) Staphylococci: Cocci cells divide in three planes in an irregular
pattern. These cells produce bunches of cocci as in grapes e.g.
Staphylococcus aureus.
• 5) Sarcinae: Sarcinae cells divide in three planes in a regular
pattern .these cells produces a cubodial arrangement of group of a
eight cells.

• Structure of Bacterial cell:

• The outer layer or cell envelope consists of two components such as
cell wall and a cytoplasmic or plasma membrane.
• Inside the plasma membrane ,there is protoplasm comprising the
cytoplasm, cytoplasmic inclusions such as
ribosome ,mesosomes,granules,vacuoles and nuclear body.
• The cell may be enclosed in a viscid layer ,which may be a loose
slime layer or organized as capsule.
• Many bacteria also posses flagella which are organs of locomotion.
 Flagella
• Flagella are long ,slender ,thin hair-like cytoplasmic
appendages ,which are responsible for the motility of bacteria.
• They are 0.01 to 0.02 µm in diameter,3 to 20 µm in length and
are found in both gram –positive and gram negative bacteria.
• These are organs of locomotion .
• Flagella can be seen by an ordinary light microscope by special
techniques in which their thickness is increased by mordanting.
• The number and arrangement of flagella are characteristic of
each bacteria.
• 1) Monotrichous: these bacteria have single polar flagellum e.g.
Vibrio cholera, Spirillum.
• 2) Lophotrichous: Bacteria have two or more flagella only at one end
of the cell.e.g. Pseudomonas fluorescens.
• 3) Amphitrichous: Bacteria have single polar flagella at both poles e.g.
Alcaligenes fecales.
• 4) Peritrichous:Several flagella present all over the surface of
bacteria .e.g. Escherichia coli, Salmonella typhi.
• Flagella are made up of a protein similar to keratin or myosin and it is
responsible for bacterial motility.
• A flagellum has three basic parts 1) Filament,2) Hook,3) Basal body.
• Filament is the thin,cylindrical,long outermost region with a constant
diameter.
• The protein in the filament is made up of monomers called ‘Flagellin’
with molecular weight ranging between 20,000 to 40,000.
• The filament is attached to a slightly wider hook, consisting of
a different protein.
• The basal body is composed of a small central rod inserted
into a series or rings.
• Gram –negative bacteria contain four rings as L-ring,P-ring,S-
ring,M-ring.
• The L-ring is embedded in lipo-polysaccharide layer of outer
membrane-ring in peptidoglycan layer,S-ring in just above
cytoplasmic membrane and M-ring within cytoplasmic
Membrane.
• Gram positive bacteria have only S&M rings in basal body.
 Pili or Fimbriae
• Pili are hair –like microfibrils,0.5 to 2µm in length and 5 to 7
nm in diameter.
• They are thinner, shorter and more numerous than flagella.
• They are present only at Gram- negative cells.
• Fimbriae are composed of protein known as pillin and its
molecular weight is 18,000 Daltons.
• They are unrelated to motility and are found on motile as well
as non-motile cells.
• They are best developed in freshly isolated strains and in
liquid cultures.
• Fimbriae or pili, these two terms are used interchangeably but they can
be distinguished.
• Fimbriae can be evenly distributed over the entire surface of the cell or
they occur at the poles of the bacterial cell.
• Pili are usually longer than fimbriae,they join bacterial cells in
preparation for the transfer of DNA from one cell to another cell and
hence, these pili are called sex pili or fertility pili.( F-Pili).
• Functions of pili:
• 1) pili play a major role in attachment to surfaces.
• 2) sex pili is used to transfer of genetic material from the donor to the
recipient cell.
• 3) many fimbriated bacteria form surface pellicles in liquid media.
• 4) Fimbriae are antigenic.
 Capsule
• Many prokaryotic microorganisms synthesize amorphous organic
exopolymers which are deposited outside the cell wall called capsules.
• The term capsule refers to the layer tightly attached to the cell wall .
• Capsule layer may be thin of a size less than 0.2µm called microcapsule
and thick layer of size more than 0.2µm to 10µm called macrocapsule.
• Capsulated bacteria produce smooth colonies on surface of agar media.
• Development of capsule is dependent on the existence of favorable
environmental conditions such as sugar concentration, blood serum or
growth in a living host.
• Capsules may composed of a complex polysaccharide or
polypeptide .water is the main component of bacterial capsule.
 Function of capsule
• 1) Capsules protect the bacteria from antibacterial
agents such as lytic enzymes.
• 2)They inhibit phagocytosis and contribute to the
virulence of pathogenic bacteria.
• 3)They may block attachment of bactriophages.
• 4)They may promote attachment of bacteria to
surfaces.
 Cell wall
• Cell wall is a rigid structure which gives definite shape to the cell wall,
situated between the capsule and cytoplasmic membrane.
• It is about 10-20 nm in thickness and constituents 20-30% of the dry
weight of the cell.
• The cell wall of a bacteria contains diaminopimelic
acid(DAP) ,Muramic acid and teichoic acid.
• These substances give rise to a complex polymeric structure known as
peptidoglycan or murein or mucopeptide.peptidoglycan consists of three
parts:
• 1) a backbone ,composed of alternating N-acetyl glucosamine and N-
acetyl muramic acid.
• 2) A set of tetrapeptide side chains attached to N-acetylmuramic acid.
• 3) A set of pentapeptide cross-bridges.
• Peptidoglycan is the major constitute of the cell wall of gram –
positive bacteria whereas in the gram-negative bacteria cell wall its
presence is only 5-10%.
• Gram positive cell wall also contain teichoic acid and
polysaccharides.
• Gram negative cell walls are more complex and the cell wall
contains lipopolysaccharide.
• The outermost layer of gram negative bacterial cell wall is called
the outer membrane, which contains various proteins known as
outer membrane proteins.
• Lysozyme enzyme present in many tissue fluids causes lysis of
bacteria .if gram positive cell is treated with lysozyme and the cell
wall is completely removed, the cell is called protoplast.
 Functions of cell wall
• Cell wall is involved in growth and cell division of
bacteria.
• It gives shape to the cell
• It gives protection to the internal structure and acts as
a supporting layer.
• It provides attachment to complement.
• It contains receptor sites for phages and colicin.
• It shows resistance to the harmful effects of
environment.
 Cytoplasmic membrane
• The cytoplasmic membrane is a thin layer lining the inner surface of the cell
wall and separating it from the cytoplasm.
• It is composed of phospholipids and proteins.
• The phospholipids form a bilayer in which most of the proteins are
tenaciously held and are called integral proteins.
• The proteins which are loosely attached and can be removed by mild
treatment such as osmotic shock are called peripheral proteins.
• Functions :
• 1) it acts as a semipermeable membrane controlling the inflow and out flow
of metabolites to and from the protoplasm.
• 2) it provides mechanical strength to the bacterial cell.
• 3) it helps in DNA replication.
• 4) It contains the enzyme ,permease ,which plays an important role in the
passage of selective nutrients through membranes.
 Cytoplasm
• The bacterial cytoplasm is a suspension of organic ,inorganic
solutes in a viscous water solution.
• The cytoplasm of bacteria differs from that of higher eukaryotic
microorganisms in not containing endoplasmic reticulum,golgi
apparatus, mitochondria and lysosomes.
• It contains the nucleus,ribosomes,proteins and other water
soluble components and reserve material.
• RIBOSOMES:
• The most notable structures in the bacterial cytoplasm are the
ribosomes.
• They are involved in protein synthesis.
• The bacterial ribosomes are referred to as 70S ribosomes .
• These ribosomes when placed in a low concentration of
magnesium, dissociate into two components as 50S &30S
particles.
• Each 50S particle contains one molecule of 23S-RNA,one
molecule of 5S-RNA and 32 different proteins.
• The 30S subunit contains one molecule of 16 r-RNA and 21
different proteins.
• NUCLEUS:
• Bacterial nucleus can be demonstrated by acid or ribonuclease
hydrolysis.
• They appear as oval or elongated bodies, generally one per cell.
• Bacterial nucleus does not possess nuclear membrane,
nucleolus and deoxyribonucleoprotein.
• The bacterial chromosome is haploid and replicates by simple
fission instead of mitosis as in an eukaryotic cell.
• A bacterial cell may possess extra nuclear genetic elements
consisting of DNA. These cytoplasmic carriers of genetic
information are termed as plasmid or episomes.
• SPORES:
• Many bacterial species produce spores inside the cell
(endospores) as well as outside the cell (exospores) e.g.
Bacillus anthracis,Clostridium tetani,Streptomyces.
• Endospores are thick-walled ,highly refractile bodies .
• Each bacterial spore, on germination forms a single vegetative cell.
• Therefore ,sporulation in bacteria is a method of preservation and not reproduction.
• Spores are extremely resistant to dessication ,staining, disinfecting chemicals and heat.
• All endospores contain large amount of Dipicolinic acid with 10 to 15 percent of the
spores being dry weight.
• The endospore consists of a core or envelope or protoplast.
• The spore envelope consists of the inner membrane ,cortex, outer membrane and spore
coat.
• The spore coat is formed outside the outer membrane and is made up of keratin-like
protein.
• The spores may be round, oval or elongated occupying a terminal ,sub terminal or
central position.
• The process of endospore formation is known as sporulation and it may take 4 to 8
hours in a vegetative cell.
 VIRUS
• Viruses are infectious agents so small that they can only be seen at
magnifications provided by the electron microscope.
• They are 10 to 100 times smaller than most bacteria.
• Virus are referred to as obligate intracellular parasites.
• Viruses largely lack metabolic machinery of their own to generate
energy or synthesize proteins.
• The viral genetic material is either DNA or RNA ,but the virus
does not have both.
• Nucleic acid is enclosed in a highly specialised protein coat.
• The structurally complete, mature and infectious virus is called the
‘virion’.
• They multiply by a complex process and not by binary fission.
• The medical importance of viruses lie in their ability to cause
a very large number of human diseases.
• Viral diseases range from minor ailments such as the common
cold to highly fatal diseases such as AIDS or yellow fever.
• Viruses multiply only in cells of particular species and thus
they are divided into three main classes:
• 1) Bacterial viruses 2) Plant viruses 3) Animal viruses.
 Classification of Viruses
• When not much was known about the structure or chemical
properties of viruses, they were classified by where they were
found.
• Thus, they have been classified as bacterial ,plant and animal
viruses.
• They have also been classified as
dermotropic,neurotropic,viscerotropic and pneumotropic
depending upon organs that they infect.
• The structure of viruses at a chemical level, a classification
has been developed based upon physiochemical properties.
Such properties are summarized as follow:
 A) Primary characteristics
• 1) Chemical nature of the nucleic acid: RNA&DNA ,single or
double stranded, single or segmented genome
strand ,molecular weight.
• 2) Structure of virion: Helical,icosahedral or complex ,naked
or enveloped,complexity,number of capsomers for icosahedral
virions,diameter of nucleocapsids for helical viruses.
• 3) Site of Replication: Nucleus or cytoplasm.
• B) Secondary Characteristics.
• 1) Host range: Host species ,specific host tissues or cell types.
• 2) Specific surface structures: e.g. antigenic properties.
• 3) Mode of transmission:e.g. feces.
 Size of Viruses
• Viruses are much smaller than bacteria. it was their small size
& filterability that led to their recognition as separate class of
infectious agents.
• As they were too small to be seen under light microscope, they
were called ultramicroscopic.
• Some of the larger viruses such as pox viruses can be seen
under the light microscope when suitably stained.
• Some of the larger viruses are about the same size of small
bacteria.
• The smallest viruses measure about 20nm.
 Structure of viruses
• A virion is a complete, fully developed viral particle composed of
nucleic acid surrounded by a coat that protects it from the environment
and serve as a vehicle of transmission from one host cell to another
host cell.
• Viruses are not cellular and therefore do not have a nucleus, cytoplasm
or cell membrane.
• Nucleic acid:
• Virus contains a single kind of nucleic acid, either DNA or
RNA ,which is the genetic material.
• The percentage of nucleic acid in relation to protein is about 1% for
the influenza viruses and about 50% for certain bacteriophages.
• The nucleic acid of a virus can be single-stranded or double stranded.
• Depending on the viruses, the nucleic acid can be linear or circular.
 Capsid & Envelope
• The nucleic acid of viruses is surrounded by a protein coat called the capsid.
• The structure of capsid is ultimately determined by the viral nucleic acid.
• Each capsid is composed of protein subunits called ‘capsomeres’.
• In some viruses, the proteins composing the capsomers are of a single type.
• In some viruses the capsid is covered by an envelope ,which usually consists of
some combination of lipids ,proteins and carbohydrates.
• Depending on the virus, envelopes may or may not be covered by spikes,
which are carbohydrate protein complexes that project from the surface of the
envelope.
• Some viruses attach to host cells by means of spikes.
• Viruses whose capsids are not covered by an envelope are known as naked
viruses .
 Morphology of viruses
• Viruses may be classified into different types, depending on
the capsid structure.
• 1) Helical viruses: helical viruses resemble long rods that may
be rigid or flexible.
• The viral nucleic acid is found within a hollow, cylindrical
capsid, which has a helical structure.
• The genetic material is generally single-stranded RNA bound
into protein helix by interactions between the negatively
charged nucleic acid and positive charges on the protein.
• E.g. Rabies virus, Tobacco mosaic virus.
• 2) Polyhedral viruses: many bacterial ,plant and animal viruses are many-
sided or polyhedral .
• The capsid of most polyhedral viruses have the shape of an
icosahedrons ,a regular polyhedron with 20 triangular faces and 12
corners.
• In icosahedral capsids the nucleic acid ,together with certain proteins, is
tightly packed in a central core, forming a pool of parallel loops around a
cylindrical hole.e.g. Adenovirus, Poliovirus.
• 3) Enveloped viruses: they are roughly spherical but highly pleomorphic
in nature.
• When helical or polyhedral viruses are enclosed by envelopes,they are
called enveloped helical and enveloped polyhedral viruses.
• The viral envelope is responsible for protection of viruses from enzymes
and certain chemicals.
• 4) Complex viruses: some viruses ,particularly bacterial viruses
have very complicated structures .these are called complex viruses.
• Certain bacterial viruses ( bacteriophages) that infect E.coli are
called T even phages .
• They are tadpole shaped, with a hexagonal head and a cylindrical
tail.
• The head consists of a tightly packed core of nucleic acid
surrounded by a protein coat or capsid.
• The size of head varies in different phages from 28nm to 100nm.
• The tail is composed of hollow core,a contractile sheath
surrounding the core and a terminal base plate which has attached
to it prongs, tail fibers or both.
 Life cycle of bacteriophages
• Bacteriophages exhibit two different types of life cycles.
• 1) Lytic cycle.
• 2) Lysogenic cycle.
In lytic cycle there is intracellular multiplication of phages
followed by lysis and release of progeny virus.
In lysogenic cycle the phage DNA becomes integrated with the
bacterial genome, replicating synchronously without any cell
lysis.
 Lytic cycle
• The replication of virulent phage was initially studied using
the T-even numbered phages of E.coli .
• The multiplication cycle of these phages like that of all
viruses, occurs in five stages.
• 1) Attachment or Adsorption
• 2) Penetration
• 3) Biosynthesis
• 4) Maturation
• 5) Release of progeny phage particles
 Attachment
• The first step in infection of a host bacterial cell by a phage is
adsorption .
• A phage attaches to the surface of a susceptible bacterium by
its tail.
• Adsorption depends on the presence of complementary
chemical groups on the receptor sites of the bacterial surface
and on the terminal base plate of the phage.
• Host specificity of phages is determined at the level of
adsorption.
 Penetration
• Attachment is followed by injection of DNA into the bacterial
cell.
• The phage DNA is injected into the bacterial body through the
hollow core .
• Penetration may be facilitated by the presence of the phage tail
of lysozyme which breaks a portion of the bacterial cell wall
for the entry of the phage DNA.
• After penetration of DNA the empty head and tail of the phage
remain outside the bacteria called ghost.
 Biosynthesis
• After infection and penetration of DNA transcription of a part of the
viral genome produces ‘early’ mRNA molecules, which are
translated into a set of early proteins.
• These serve to switch off host cell macromolecular
synthesis ,degrade the host DNA and start to make the components
of viral DNA.
• The viral DNA replicates and also start to produce a batch of mRNA
• Maturation:
• Phage DNA ,head and tail protein are synthesized separately in the
bacterial cell . The DNA is considered into a compact polyhedron
and packaged into the head and finally, the tail structure are added.
The process of assembly of the phage from its component is called
maturation.
 Release of progeny phage particle
• The release of progeny phage particles takes place by sudden
explosion or brusting of the bacterial cell wall.
• Lysozyme synthesized within the cell causes the bacterial cell
wall to break down and the newly produced bacteriophages
are released from the host cell.
• The released bacteriophages infect other susceptible bacterial
cells.
• Each cycle of phage reproduction may require 20 to 60 mins .
• The sequence of events by the injection of the phage nucleic
acid and release of newly synthesized virions is called
‘replication cycle’
 Lysogenic cycle
• In contrast to virulent phages, temperate phage don’t cause
lysis of the host cell.
• Following entry into the host cell, the temperate phage nucleic
acid become integrated with the bacterial chromosome.
• The integrated phage nucleic acid is known as the prophage.
• The prophage behaves like a segment of the host chromosome
and replicates synchronously in the bacterial cell. this is called
lysogeny and a bacterium that carries a prophage within its
genome is called lysogenic bacterium.
• The prophage confers certain new properties on the lysogenic
bacterium. This is known as ‘lysogenic conversion’.
• A lysogenic bacterium is resistant to
reinfection by the same or related phages .
• This is known as ‘superinfection immunity’
 Fungi
• Fungi are a group of non-motile eukaryotic organisms which
exist as saparophytes,parasites or commensals.
• They posses differentiated nuclei surrounded by a nuclear
membrane and reproduce either by budding or by forming
spores.
• They have rigid chitinous cell wall.
• All fungi are chemoheterotrophs,requiring organic compounds
for energy and carbon.
• Fungi are aerobic and facultative anaerobic.
• The majority of fungi are saprophytes in soil and water.
• The study of fungi is called mycology.
 Classification of Fungi
• Fungi are placed in the phylum Thallophyta which contains the
irregular plant masses that lack definite root, stem and leaf
structures.
• Depending on cell morphology ,fungi can be divided into 4 classes:
1)moulds,2)yeasts,3)yeast like fungi and 4)dimorphic fungi.
• 1) Moulds:
• Fungi which form mycelia are called moulds .
• The thallus of a mould consists of long filaments of cells joined
together, these filaments are called hyphae.
• In most mould ,the hyphae contain crosswalls called septa,which
divide the hyphae into distinct,uninucleate or multinucleate cell-
like units.
• These hyphae are called septate hyphae.
• In a few classes of fungi ,the hyphae does not contain septa and
appear as long,continous cells with many nuclei. these hyphae
are called coenocytic or non-septate hyphae.
• When environmental conditions are suitable ,the hyphae grow
and form a mass called mycellium.
• E.g. : Aspergillus niger,Penicillum notatum
• 2) Yeasts:
• Yeasts are round,oval,or e4longated unicellular fungi.
• Most of them reproduce by an asexual process called budding in
which the cell develops a protuberance which enlarges and
eventually separates from the parent cell.
• On culture, they form smooth, creamy colonies e.g.
Saccharomyces cerevisae,Cryptococcus neoformans.
• Difference between Mould and yeast:
 3) Yeast like fungi
• In some yeasts like Candida albicans ,the bud remains attached to
the mother cell and elongates, followed by repeated
budding ,forming chains of elongated cells known as pseudohyphae.
• These can be differentiated from true hyphae because they have a
constriction at the septa are also present at the branching point.
• 4) Dimorphic fungi: some fungi, mainly pathogenic species exhibit
dimorphsim,i.e. two forms of growth.
• Such fungi can grow either as a mould or as a yeast.
• The mould like forms produce vegetative and aerial mycelium.
• The yeast like forms reproduce by budding.
• At 37 degree C ,the fungus grows yeast like and at 25 degree C it
shows mould like growth.eg. Mucor rouxii.
• The systematic classification of fungi: Based on their sexual
spore formation, fungi are divided into 4 classes.
• A) Phycomycetes: they are fungi having non-septate hyphae.
• Phycomycetes also produce sexual spores known as oospores
and zygospores e.g. Mucor,Rhizopus etc.
• B) Ascomycetes: they form sexual spores within a sac and are
called ascopers.
• The sac is called ascus.they form septate hyphae.e.g.
Histoplasma,Candida etc.
• C) Basidiomycetes : they reproduce sexually and form septate
hyphae. These basidiospores are borne at the tip of the basidium
e.g. Cryptococcus neoformans.
• D) Fungi imperfecti : they are also called Deuteromycetes or
Hyphomycetes .they consist of groups of fungi whose sexual
phases have not been identified.
• Many imperfect fungi form septate hyphae and asexual conidia.
• A majority of the pathogenic moulds,yeasts,yeast-like fungi and
dimorphic fungi belong to the group fungi ‘imperfecti’ e.g.
Trichophyton,Epidermophyton.
• Importance of fungi:
• 1) Fungi are important source of antibiotics. E.g. Penicillin.
• 2) Yeast and moulds are good sources of different enzymes e.g.
amylase produced from Aspergillus species.
• 3) fungi have been used to alter the texture, improve the
flavour.
• 4) edible wild or domesticated varieties of mushrooms are
important as food sources.
• 5) Yeast are used for fermentation purposes in production of
beverages and juices.
• 6) Fungi have capability to breakdown com
Reproduction of Fungi
 Rickettsia
• Rickettsia are small ,gram-negative bacilli which have
properties in between bacteria and viruses.
• They are primarily parasites of arthropods such as fleas,
lice ,mites and ticks.
• They infect the vascular endothelium and reticuloendothelial
cells in vertebrates ,including human beings.
• The family Rickettsiaceae is named after Howard Taylor
Ricketts.
• Who discovered the spotted fever rickettsia in 1909.
 General characters
• 1) the organisms of Rickettsiae are small coccobacilli .
• 2) they are obligate intracellular parasites.
• 3) the multiply by binary fission
• 4) they possess both DNA and RNA.
• 5) they are sensitive to lysozyme and antibiotics.
• 6) they contain muramic acid in the cell wall.
• 7) they produce only endotoxin and exotoxin.
• 8)they require an arthropod vector as a part of their natural
cycle.
• 9) they possess a trilaminar cytoplasmic membrane.
 Cultural characteristics
• They are unable to grow in cell free media except Rochalimaea
quintana which grows on blood agar.
• Growth generally takes place in the cytoplasm of infected cells.
but in the case of spotted fever rickettsiae ,growth may take
place in the nucleus.
• They are cultivated in the yolk sac of a developing chick
embryo.
• They may also grow on chorioallantoic membrane.
• Optimum temperature of incubation for rickettsiae is 35 degree
C and for those of the spotted fever group is 33 degree C.
• After incubation ,the eggs are harvested and the yolk sac
membrane is removed.
• They also serve as an excellent source of organisms for
diagnostic antigens and vaccines.
• Rickettsial growth is enhanced in the presence of sulfonamides
and rickettsial diseases are made more serve by these drugs.
• Para-amino benzoic acid inhibits the growth of rickettsial
organisms.
• Pathogenesis:
• Rickettsiae are transmitted to man by the bite or faeces of an
infected arthropod vector.
• On entry into a human body,they multiply locally and enter the
blood stream followed by multiplication of the organisms in the
endothelial cells lining of the small blood vessels.
• Rickettsiae exhibit two types of toxicity .
• They possess an endotoxin that kills mice following an
intravenous injection.
• Endotoxin activity is specifically neutralized by antiserum.
• Diseases caused by various rickettsial species are typhus fever,
spotted fever,scrub typhus,Q-fever and trench fever.
 Spirochetes
• The spirochetes form one of the major phyla of the kingdom of
Eubacteria.
• The length between 3-500 pm and diameters around 0.09 to 3 pm.
• The spirochetes are flexible and are of either spiral or helical curved
shaped.
• Spirochetes are heterotrophic organisms i.e they do not prepare their
own food material and depend on other organic matter for their
survival.
• They are motile but without flagella. The motility is due to the
presence of axial fibres or internal pseudo flagella within the cell
wall.
• Spirochetes are very minute and are invisible even in normal
microscope.
• Dark film microscopy and phase contrast microscopy are used to
observe spirochetes.
• The cell wall is thin and two layered –the outer membrane and the inner
P- layer ( Peptidoglycan layer)
• In between these two layers a periplasmic area is present in which axial
fibres or internal pseudo flagella are present.
• These reproduce through asexual transverse binary fission. Spirochetes
are free living, anaerobic organisms organisms and pathogenic in nature.
• The taxonomical classification for spirochetes is they belong to the
order Spirochaetales and families Brachyspiraceae,Leptospiraceae and
Spirochaetaceae.
• The pathogenic diseases caused by spirochetes are Leptospirosis,Lyme
disease and relapse fever and Syphilis .
 IMP Questions

• 1) Different methods used for identification of

microbes
• 2) Draw the ultra structure of bacteria and explain it
in brief.
• 3)Life cycle of virus (Lytic & Lysogenic)
• 4) Classification of fungi .( short que)
• 5) Reproduction of fungi.
• 6) Rickettsia ( only read once ,if they ask)