Dr.

Nurjahan Begum
Assistant Professor
Department of Microbiology
DCIMC
Bacterial pathogenesis
Determinants of pathogenicity
Endotoxin, Exotoxin
Koch’s postulates
 Basic terms frequently used in describing
aspects of pathogenesis
Infection: Multiplication of an infectious agent within the
body (Multiplication of the bacteria that are part of normal
flora of gastrointestinal tract, skin, etc, is generally not
considered an infection).

Pathogenicity islands: Pathogens can be distinguished

from their avirulent counterparts by the presence of specific
gene clusters in the genome known as pathogenicity islands.

Pathogenicity: The ability of an infectious agent to cause

disease.
 Basic terms frequently used in describing
aspects of pathogenesis
Virulence: The quantitative ability of an agent to cause disease
and is measured by the number of organism require to cause
disease.
Toxigenicity: The ability of a microorganism to produce a toxin
that contributes to the development of disease.
Invasion: The process whereby bacteria, parasites, fungi and
viruses enter the host cells or tissues and spread in the body.
Pathogen: A microorganism capable of causing disease.
Non-pathogen: A microorganism that does not cause disease. It
may be part of the normal flora.
 Basic terms frequently used in describing
aspects of pathogenesis
Opportunistic pathogen: An agent capable of causing
disease only when the host’s resistance is impaired (e.g. the
patient is immunocompromised).
Colonization: Refers to the presence of a new organism that
is neither a member of normal flora nor the cause of
symptoms.
50% Lethal dose (LD50):The number of organism needed to
kill half of the hosts is called LD50.
50% infectious dose (ID50): The number of organism
needed to cause infection in half of the hosts is called ID50.
 Basic terms frequently used in describing
aspects of pathogenesis
Endemic: The infection that occurs at a persistent,
usually low level in a certain geographical area is
called endemic.

Epidemic: The infection that occurs at a much higher

rate than usual is known as epidemic.

Pandemic: Infection that spreads rapidly over large

areas of the world is known as pandemic.
 Types of infections
Primary infection: This condition denotes an initial
infection with an organism in a host.
Reinfection: This condition denotes subsequent infection
with the same organism in the same host.
Secondary infection: This condition denotes an infection
with a new organism in a host whose body resistance is
already lowered by a pre-existing infectious disease.
Cross-infection: This condition denotes an infection with a
new organism from another host or another external source
in a patient who is already suffering from a disease.
 Types of infections contd…
Nosocomial infection: Cross-infections acquired in hospitals
are called hospital-acquired or nosocomial infections.
Iatrogenic infection: This condition denotes a physician
induced infection as a result of therapy with drugs or
investigation procedures.
Inapparent/Subclinical infection: Inapparent clinical
infections are called subclinical infections.
Latent infections: This denotes a condition in which some
organisms may retain in a latent or hidden stage in host and
subsequently they multiply to produce clinical disease when
host resistance is lowered.
 Pathogenesis
Mechanism of development of disease is called pathogenesis.
Stages of pathogenesis:
 Transmission of infection
 Evasion of primary host defense
 Adherence to cell surface
 Colonization by growth of the bacteria at the site of adherence.
 Disease symptoms caused by toxin production or by invasion
accompanied by inflammation.
 Host responses both specific & non-specific.
 Progression or resolution of disease.
 Transmission of infection

There are three important components that play an

important role in successful transmission of microbial
disease. These are
Reservoir
Mode of transmission
Susceptible host
 Transmission of infection contd…
Reservoir: Reservoirs are human, animal, plant, soil, or
inanimate matter in which organisms usually live, multiply,
and cause the infections with or without overt clinical
manifestations.
Carrier: A carrier is a person who harbors pathogenic
microorganisms without showing any signs and symptoms
of disease.
Zoonotic infections: Infections transmitted from animals
to humans are called zoonotic infections. e.g. Plague,
Anthrax, Intestinal TB, Brucellosis, Leptospirosis, Lyme
disease, Psittacosis, Rocky Mountain spotted fever, Q fever,
Tularemia.
 Determinants of pathogenicity
(virulence factors)
1. Mode of transmission
2. Adherence factors
3. Invasiveness
4. Antiphagocytic factors
5. Intracellular servival
6. Antigenic heterogenicity
7. Toxins
 Mode of transmission
Horizontal Pathogen Disease
transmission
Faeco-oral route Shigella dysenteriae Bacillary dysentery
Salmonella typhi Typhoid fever
Vibrio cholerae Cholera

Respiratory route Mycobacterium tuberculosis Tuberculosis

Neisseria meningitidis Meningitis
Streptococus pneumoniae Pneumonia
Haemophilus influenzae Meningitis
Skin Clostridium tetani Tetanus

Sexual contact Treponema pallidum Syphilis

Neisseria gonorrhoeae Gonorrhoea
Chlamydia trachomatis Urethritis
 Mode of transmission
contd…
Vertical Pathogen Disease
transmission

Transplacental Treponema pallidum Congenital syphilis

Listeria monocytogens Neonatal sepsis & meningitis

During passage Streptococcus agalactiae Neonatal sepsis & meningitis

through birth Neisseria gonorrhoeae Ophthalmia neonatorum
canal Chlamydia trachomatis Pneumonia or conjunctivitis

Breast milk Staphylococcus aureus Skin or oral infection

 Adherence factors
Bacterial adherence to the cell surface is mediated by
various molecules known as “adhesins”. These are
Pili
Capsule
Glycocalyx/Slime layer
Lipoteichoic acid
 Invasiveness
Refers to the ability of an organism to invade the host
cells after establishing infection.
Invasiveness is facilitated by several bacterial
enzymes such as
Coagulase
Collagenase
Hyaluronidase
IgA protease
Streptokinase (fibrinolysin)
Lecithinase
 Enzymes
Coagulase: Produced by staphylococcus aureus. It is of
two types
▪ Bound coagulase
▪ Free coagulase

 Bound coagulase causes deposition of fibrin on the

surface of Staphylococci. Thus protect the bacteria from
phagocytosis.
 Free coagulase forms fibrin barrier surrounding the
bacteria thus prevent the entry of phagocytic cells & also
causing localized infection.
 Enzymes

Hyaluronidase: Continue…..
 Produced by Streptococci, Staphylococci & anaerobes.
 It hydrolyses hyaluronic acid which is a ground substance
of connective tissue, thus helping in spreading of infection.

Streptokinase:
 Produced by Streptococcus pyogenes.
 Activate proteolytic enzyme of plasma which dissolves
coagulated plasma & probably aid in the rapid spread of
Streptococci.
 Streptokinase has been used in the treatment of acute MI to
dissolve fibrin clots.
 Enzymes
Collagenase:
Continue…..
 Produced by Clostridium perfringens
 Degrades collagen & promotes spread of infection.
Lecithinase:
 Produced by Clostridium perfringenes
 Hydrolyzes lecithin in the cell membrane, resulting in destruction
of the membrane and widespread cell death.
IgA1 protease:
 Produced by Strep. pneumoniae, N. gonorrhoeae, N. meningitidis,
H. influenzae.
 They split IgA1 & inactivate its antibody activity, thus hampering
mucosal immunity.
 Antiphagocytic factors
Capsule: Prevent phagocytosis of bacteria by prevent C3b
deposition on the surface of bacteria. Also negative charges on
capsular polysaccharide repel the negatively charged
neutrophil.
Protein A: Bind with the Fc portion of IgG at the complement
binding site. So prevent complement activation. No C3b is
produced & prevention of opsonization & phagocytosis.
M protein: Interfere with the deposition of complement
component C3b on to the Streptococcal cell surface. So,
activation of alternative complement pathway & opsonization
of Streptococcus is inhibited.
Pili: They prevent phagocytosis by antigenic variation.
Capsule
 Intracellular survival
A few mechanism that are suggested for intracellular survival
of bacteria include
 Inhibition of fusion of phagosome with lysosome
 Inhibition of acidification of the phagosome
 Escape from phagosome into cytoplasm.

Example:
 Facultative intracellular: Mycobacterium, legionella, listeria,
Brucella.
 Obligate intracellular: Chlamydia, Rickettsia.
 Antigenic heterogenicity
Some bacteria have ability to make frequent shift in
antigenic form thus help to evade immune system. e.g.
Neisseria gonorrhoeae.
 Toxins
Toxin produced by bacteria are generally divided into
two groups:

a. Exotoxins
b. Endotoxins
Properties Exotoxin Endotoxin

Source Gram (+) ve & Gram (-)ve bacteria Cell wall of Gram (-)ve bacteria

Nature Protein (Polypeptide) Lipopolysaccharide

Location of genes Plasmid or bacteriophage Chromosome

Nature of Actively secreted by bacteria Not secreted by the bacteria

secretion

Toxicity High Low

Heat stability Heat labile (except Staphylococcal Heat stable

enterotoxin & heat stable toxin of E.
coli)
Mode of action Mostly enzyme like action Mediated by IL-1 & TNF

Clinical effects Specific pharmacological effect Non-specific (fever, shock etc)

Antigenicity Highly antigenic Poorly antigenic

Vaccines Toxoid used as vaccine No toxoid, no vaccine

 There are 3 types of Exotoxins
1. Cytotoxins: kill cells e.g. Diphtheria toxin

2. Neurotoxins: interfere with normal nerve impulses.

e.g. Botulinum Toxin

3. Enterotoxins: effect cells lining the GI Tract. e.g.

Cholera toxin or choleragen.
 Mechanism of exotoxins

A. By activation of adenylate cyclase

B. By blocking protein synthesis
C. By acts as superantigen
D. By acts as protease
 Mechanism of exotoxins
A. By activation of adenylate cyclase
↑cAMP →as a consequence cAMP dependent protein
kinase activity increased, an enzyme that
phosphorylates ion transporters in the cell membrane
→ as a result fluid & electrolytes comes out from the
cell & also prevent reabsorption of some electrolytes
& H2O.
e.g. Cholera toxin
Heat-labile toxin of E. coli
Pertussis toxin
Edema factor of anthrax toxin
M/A of Cholera toxin
 Mechanism of exotoxins cont…
B. By blocking protein synthesis
 By ADP ribosylation of EF2 → No elongation of peptide
chain → Block protein synthesis → Cell death.
e.g. Diphtheria toxin, Exotoxin A of Pseudomonas

 By removing adenine base from 28S rRNA at position

4324 in the 60S unit of human ribosome → irreversible
inactivation of 60S ribosome → Block protein synthesis
→ Cell death.
e.g. Shiga toxin, Verotoxin/Shiga-like toxin producing E.
coli.
 Mechanism of exotoxins
cont…
C. By acts as a superantigen
Toxin acts as a superantigen → Directly binds with many T-
cell at a time without processing → T-cell activated →
Produce cytokines.
e.g.
TSST of Staphylococcus aureus & Streptococcus pyogens
Staphylococcus aureus enterotoxin
Streptococcus pyogens erythrogenic toxin
Bacillus cereus enterotoxin
 Mechanism of exotoxins
cont…
D. By acts as Protease
e.g. Exfoliatin/epidermolytic toxin of Staph. aureus
Tetanus toxin
Botulinum toxin
Lethal factor of anthrax toxin

Exfoliatin: is a protease that cleave despmoglein in

desmosomes of skin, as a result detachment of
epidermis at the granular cell layer.
 Mechanism of exotoxins
cont…
Tetanus toxin: Light chain is a protease that cleave the
proteins responsible for release of inhibitory
neurotransmitters (GABA, glycine).
Botulinum toxin: Light chain is a protease that cleave
the proteins involved in acetylcholine release.
Lethal factor of anthrax toxin: is a protease that cleave
phosphokinase required for signal transduction
pathway that controls cell growth & consequent cell
death.
 Name of some exotoxins released from
Gram positive bacteria
Name of toxin Mechanism of action Disease

Staphylococcal Acts as superantigen, causes polyclonal T cell Food

enterotoxin activation thus releasing large amount of IL poisoning
which stimulate the enteric nervous system to
activate the vomiting center in the brain leading
to profound vomiting.

Staphylococcal Acts as superantigen, causes polyclonal T cell Toxic shock

TSST activation thus releasing large amount of IL syndrome
which causes S/S of toxic shock.

Exfoliatin of Protease in nature, cleaves desmoglein in Scalded

staph. aureus desmosomes, leading to the separation of skin
The epidermis at the granular cell layer. syndrome
Name of toxin Mechanism of action Disease

Erythrogenic toxin Acts as superantigen (M/A is similar to that of Scarlet fever

of Strep. pyogens TSST of Staph. aureus)

Diphtheria toxin of It has two domains A & B Diphtheria

C. diphtheriae B causes binding of exotoxin to specific
receptors on the membrane of human cell.
A is the active or toxic part which causes ADP
ribosylation of EF2.
EF2 + NAD→EF2-ADP-ribose + Nicotinamide
This reaction causes cessation of protein
synthesis of eukaryotic cell leading to cell death.

Tetanospasmin of Acts as neurotoxin. It has 2 chain. Heavy chain Tetanus

Clostridium tetani bind to ganglioside receptor of neuron, light
chain is a protease that degrades the protein
responsible for release of inhibitory
neurotransmitters leading to convulsive
contraction of voluntary muscles.
Name of toxin Mechanism of action Disease

Botulinum Acts as neurotoxin. It has 2 units. Botulism

toxin of One for binding & one for toxicity.
Clostridium Toxic part is protease in nature which
botulinum degrades protein responsible for release of
acetylcholine at peripheral nerve synapses,
leading to flaccid type of paralysis.

Alpha toxin of Lecithinase in nature. It hydrolyzes lecithin Gas gangrene

Clostridium in the cell membrane resulting in destruction
perfringens of the membrane leading to cell death.

Exotoxins of Exotoxin A is an enterotoxin that causes Pseudomembranous

Clostridium watery diarrhoea. Exotoxin B is a cytotoxin colitis
difficile that damage colonic mucosa and produce
pseudomembrane. Both exotoxin A &
exotoxin B glycosylate signal transduction
proteins resulting disaggregation of actin
filaments in cytoskeletal integrity, apoptosis
& death of enterocytes.
Name of toxin Mechanism of action Disease

Exotoxins of Protective antigen: Binds with cell Anthrax

Bacillus surface & forms pore through which
anthracis edema factor & lethal factor enter into the
cell.

Edema factor: is an adenylate cyclase

that raises cAMP within cell, leads to loss
of chloride ion & water and consequent
edema formation in tissue.

Lethal factor: Protease in nature that

degrades phosphokinase required for
signal transduction. Thus hampering cell
growth and consequent cell death.
Name of Mechanism of action Disease
toxin
Exotoxins of Two enterotoxins. Food poisoning
Bacillus One enterotoxin acts same as cholera
cereus toxin which causes ADP ribosylation of
stimulatory coupling protein (Gs protein)
leading to stimulation of adenylate cyclase
that increases caMP, resulting in excretion
of chloride & H2O from cell & inhibition of
Na absorption.

Another enterotoxin acts as superantigen

resembles that of staphylococcal
enterotoxin (causes polyclonal T cell
activation thus releasing large amount of IL
which stimulate the enteric nervous system
to activate the vomiting center in the brain
leading to profound vomiting.
 Name of some exotoxins released from
Gram negative bacteria
Name of Toxin Mechanism of action Disease
Heat labile toxin ADP ribosylation of stimulatory coupling Watery diarrhoea
of E. coli protein (Gs protein) leading to stimulation of
adenylate cyclase that increase cAMP
resulting in excretion of chloride & H2O
from enterocyte into the lumen & inhibition
of Na absorption.

Heat stable toxin Stimulate guanylate cyclase thus increase Watery diarrhoea
of E. coli concentration of cyclic GMP which inhibit
reabsorption of Na ion.
Shiga toxin B subunit bind to the ganglioside receptors Bacillary
of enterocyte. A subunit inhibit protein dysentery
synthesis by removing adenine base from
28S rRNA at position 4324 in the 60S unit
of human ribosome
Name of Mechanism of action Disease
toxin
Cholera ADP ribosylation of stimulatory coupling protein Cholera
toxin of (Gs protein) leading to stimulation of adenylate
Vibrio cyclase that increase cAMP, resulting in excretion
cholerae of chloride & H2O from cell & inhibition of Na
absorption.

Pertussis ADP ribosylation of inhibitory G protein (Gi Pertussis

toxin of protein) that controls adenylate cyclase activity
Bordetella leading to stimulation of adenylate cyclase activity
pertussis resulting in increased amount of cAMP causing
edema.

Another effect is inhibition of signal transduction,

resulting inability of lymphocytes to migrate to and
enter lymphoid tissue. So, increase lymphocyte in
blood and causes marked lymphocytosis.
 Mechanism of action of exotoxins
at a glance
Mechanism of Exotoxin
action
ADP-ribosylation Diphtheria toxin (of EF2), Cholera toxin (of Gs protein),
E. coli heat-labile toxin (of Gs protein), Pertussis toxin
(of Gi protein)

Superantigen Toxic shock syndrome toxin of Staph. aureus & Strep.

Pyogenes, staphylococcal enterotoxin, erythrogenic
toxin, enterotoxin of B. cereus & enterotoxin of Cl.
Perfringens
Protease Exfoliatin, scalded skin toxin, tetanus toxin, botulinum
toxin, lethal factor of anthrax toxin

Lecithinase Clostridium perfringens alpha toxin

 Exotoxins that increase cAMP/Stimulate
adenylate cyclase activity
Bacteria Exotoxin Mode of action

Vibrio cholerae Cholera toxin ADP ribosylates Gs protein, which

activates it, thereby stimulating
adenylate cyclase.

E. coli Heat labile toxin Same as cholera toxin

Bordetella Pertussis toxin ADP-ribosylates Gi protein, which

pertussis inactivates it, therby stimulating
adenylate cyclase

Bacillus Edema factor of Is an adenylate cyclase

anthracis anthrax toxin
Toxins encoded by lysogenic bacteriophage
Enterotoxin of Staphylococcus aureus
Erythrogenic toxin of Streptococcus pyogenes
Exotoxin of Corynaebacterium diphtheriae
Exotoxin of Clostridium botulinum
Exotoxin (enterotoxin) of Vibrio cholerae
Exotoxin (enterotoxin) of Shigella dysentery
 Lysogenic conversion is the term applied to the new
properties that a bacterium acquires as a result of
expression of the integrated prophage genes.
 Endotoxin
It is an integral part of cell wall
of Gram (-)ve bacteria.
It is located on the outer
membrane of cell wall of gram
(-)ve bacteria.
Chemical nature is
lipopolysaccharide
Encoded by genes on the
bacterial chromosome
Toxicity retain in the lipid part.
Mode of action of endotoxin
 Biologic effects of endotoxin
Fever due to the release of endogenous pyrogen
(interleukin-1,6) from macrophage which acts on the
hypothalamic temperature-regulatory center
Hypotension, shock, and impaired perfusion of essential
organs owing to bradykinin-induced vasodilation, TNF-
induced increased vascular permeability, and decreased
peripheral resistance (nitric oxide, a potent vasodilator,
also causes hypotension)
 Biologic effects of endotoxin
Disseminated intravascular coagulation (DIC) due to
activation of the coagulation system through Hageman factor
(factor XII), resulting in thrombosis, a petechial or purpuric
rash, and tissue ischemia, leading to failure of vital organs
Activation of the alternative pathway of the complement
cascade, resulting in inflammation and tissue damage.
Activation of macrophages, increasing their phagocytic
ability, and activation of many clones of B lymphocytes,
increasing antibody production. (Endotoxin is a polyclonal
activator of B cells, but not T cells.)
 Biologic effects of endotoxin
Clinical findings Mediator or mechanism

Fever Interleukin 1,6

Hypotension Bradykinin and nitric oxide

(shock)

Inflammation Alternative pathway of complement (C3a, C5a)

Coagulation Activation of Hageman factor

(DIC)
 Septic shock Vs Endotoxic shock
Septic shock Endotoxic shock

Bacteria present in blood. Toxin present in blood

Blood cultures are usually Blood cultures are usually

positive negative
 Stages of an infectious disease
A. Incubation period: The time between the acquisition of
the organism (or toxin) and the beginning of symptoms.

B. Prodrome period: During which non-specific symptoms

such as fever, malaise and loss of appetite occur.

C. Specific illness period: During which the overt

characteristic signs and symptoms of disease occur.

D. Recovery period: During which the illness subsides and

the patient returns to the healthy state.
It can be difficult to show that a specific bacterial species
is the cause of a particular disease.

In 1884, Robert Koch proposed a series of postulates.

These postulates establishes the etiological relationship

between an organism and a particular diseases.
Koch´s postulates are summarized as follows:
1. The microorganism should be constantly associated with
the disease and should be always found in the diseased
person and not in healthy ones.

2. The microorganism should be grown in pure culture from

the lesion.

3. When such a pure culture is inoculated into healthy

person/susceptible animal species, the typical disease must
result.

4. The same microorganism must again be isolated from the

lesions of inoculated animal or person.
 Limitations of Koch’s postulates
1. Genetic, auto immune and noncurable microbial diseases.
Because in this cases causative agents cannot be
identified.

2. Some organisms cannot be cultured in artificial media,eg:

Treponema pallidum, Mycobacterium leprae etc.

3. Some organisms cannot produce disease in experimental

animals.eg: Neisseria gonorrhoeae

4. Some diseases may be caused by mixed infection with

more than one microbes.
 Molecular Koch’s postulates
The phenotype or property should be associated with
the pathogenic strains of microorganisms. Not with
the non pathogenic strain.

The specific inactivation of the gene associated with

virulence should lead to decrease in pathogenicity.

 Replacement of mutated gene with wild type gene

should restore the full pathogenicity to the organism.