Review on General

Bacteriology
Prof. Dr. Ripon Barua
MBBS, M.Phil (Microbiology), FCPS (Medicine)
MPH (Epidemiology), MMEd, BCS (health)
Professor of Microbiology (cc)
Chittagong Medical College, Chattogram 4203, Bangladesh.
Cell Phone and WhatsApp: +8801629881825
E-mail: riponbarua38cmc@gmail.com
ORCID: https://orcid.org/0000-0001-9898-7306
 Characteristic Prokaryotic Eukaryotic
Bacterial Human
Cells Cells
Nuclear membrane No Yes
Sterol in cell membrane No Yes
Mitotis No Yes
DNA + Histones No Yes
Chromosome number 1 >1
Membrane-bound No Yes
(mitochondria,lysosomes)
Size of ribosome 70S 80S
Peptidoglycan in cell wall Yes No
CLASSIFICATION OF BACTERIA:
A. Rigid thick-walled cell
I. Free living (extracellular bacteria)
II. Obligate intracellular bacteria

B. Flexible, thin walled cell

C. Wall-less cell
Classification contd.

Free living (extracellular bacteria)

• GRAM POSITIVE
• COCCI -- Streptococcus
-- Staphylococcus
• BACILLI
-- Sporulating - Aerobe: Bacillus
- Anaerobe: Clostridium
-- Non-sporulating
Corynebacterium, Actinomyces

• GRAM NEGATIVE
• COCCI -- Neisseria
• BACILLI -- Escherichia, Salmonella, Pseudomonas
• Acid-fast bacilli
Mycobacterium
Classification contd.

Obligate intracellular bacteria

1. Rickettsia
2. Chlamydia
Classification contd.

Flexible, thin walled cell

(spirochetes)

1. Treponema
2. Borrelia
3. Leptospira
Classification contd.

Wall-less cell

Mycoplasma
Bacterial Shapes

8
 Essential structures of Bacteria:
⚫ Cell wall
⚫ Cytoplasmic
membrane
⚫ Ribosome
⚫ Nucleoid
⚫ Mesosome
⚫ Periplasm
Non-essential structures of Bacteria:
⚫ Capsule
⚫ Appendages-
Pili, Flagella
⚫ Spores
⚫ Plasmid
⚫ Granule
⚫ glycocalyx
Cell envelop
 Peptidoglycan structure: Escherichia coli (A) has a different cross-link from that of Staphylococcus aureus (B). In E. coli, c is
cross-linked directly to d, whereas in S. aureus, c and d are cross-linked by five glycines. However, in both organisms the
terminal D-alanine is part of the linkage. M, muramic acid; G, glucosamine; a, L-alanine; b, D-glutamic acid; c, diaminopimelic
acid (A) or L-lysine (B); d, D-alanine; x, pentaglycine bridge.
 Functions of Cell Wall
⚫ Maintaining the cell's characteristic shape
⚫ Countering the effects of osmotic pressure
⚫ Providing attachment sites for bacteriophages
⚫ Providing a rigid platform for flagella, pili
⚫ Play an essential role in cell division
⚫ Major antigenic determinants of the cell
surface
⚫ Resistance of Antibiotics
Medically Important Bacteria that
Cannot Be Seen in the Gram Stain:
1. Mycobacteria, including M. tuberculosis
2. Treponema pallidum
3. Mycoplasma pneumoniae
4. Legionella pneumophila
5. Chlamydiae, including C. trachomatis
6. Rickettsiae
Functions of Cell Membrane:
1. Active transport of molecules into the cell
2. Energy generation by oxidative
phosphorylation
3. Synthesis of precursors of the cell wall
4. Secretion of enzymes and toxins
5. Bear receptor for chemotaxis
 Capsule
• polysaccharide

Anti phagocytic

it forms part of
biofilms.
Importance of Capsule:
1. Virulence factor

2. Identification of micro organism

3. Active immunization

4. Role in bacterial pathogenesis

Capsulated Bacteria:
⚫ Streptococcus pneumoniae
⚫ Neisseria meningitidis
⚫ Haemophilus influenzae
⚫ Klebsiella pneumoniae
⚫ Salmonella typhi
⚫ Yersinia pestis
⚫ Bordetella pertussis
⚫ Bacillus anthracis
⚫ Pseudomonas aeruginosa
 Ordinary pili
Organ of attachment

Sex pili
⚫ Takes part in genetic transfer
during conjugation

Antigenic
Bacteria with Pili:
⚫ Neisseria
⚫ EPEC
⚫ Salmonella
⚫ Shiigella
⚫ Klebsiella
⚫ Proteus
⚫ Streptococcus
Mechanisms of bacterial
genetic material transfer:
- Conjugation
- Transduction
- Transformation
Process of Conjugation
Transduction-bacteriophage
Life cycles –
lytic - lysis of the cell- virulent phage
Transduction-bacteriophage
Life cycles-
lysogenic - not lysis - phage DNA is integrated - temperate
phage - after many generation - induction, conversion
Lysogenic Conversion
Phage DNA itself is the new genetic element.
Bacteriophages – 2 Types of life cycle
⚫ Lytic or virulent cycle – progeny viruses
build up inside host bacterium, which rupture
to release them
⚫ Temperate or nonlytic or lysogenic cycle
– host bacterium is unharmed.
Transformation (Griffith, 1928)
⚫ Transfer of genetic information by
free DNA. i.e. by direct uptake of
donor DNA by the recipient DNA.
 Flagella
⚫ Thread like appendiges composed of flagellin protein
⚫ Organ of locomotion
⚫ 1-20/cells
⚫ Originate in protoplasm
⚫ On the basis of arrangement:
Monotrichous

Lophotrichous

Amphitrichous

Peritrichous
 Monotrichous (Vibrio) Amphitrichous (Alkaligens faecalis)

Lophotrichous (H. pylori) Peritrichous (E. coli) 30

 Spore
⚫ A resting state of bacteria
produced during
unfavourable condition &
is highly resistant to –
⚫ desiccation
⚫ heat &
⚫ Chemical agents

Marked resistance is due to-

•Dehydrated state
•Calcium dipicolinate
•Stabilization of spore enzymes
•Very low metabolic activity
Bacterial Spore
 Endospores (spores)

• Central spore: Bacillus anthracis

• Subterminal spore: Clostridium perfringens
• Terminal spore: Clostridium tetani
Sporulation
Bacterial Growth – Binary fission
 Generation time/ (Doubling time)
⚫ The time required for a population of
bacteria to double in number.
⚫ Escherichia coli (E. coli): 20 minutes
⚫ Mycobacterium tuberculosis: 20 hours
⚫ Mycobacterium leprae: 20 days
 Clinical significance

Synthesis of Exotoxin,
antibiotic, metach granule,
start of sporulation

Endotoxin,
Diph toxin,
Penicillin & sporulation
other CW
synthesis
inhibitors

Detergent
Oxygen and growth
Staining and Cluture
media
 REAGENTS USED IN GRAM STAIN
❖ Primary stain----- CRYSTAL VIOLET 1 min

❖ Mordanting-------- GRAM IODINE 1 min

❖ Decolorizer--------Acetone + Methanol
❖ Counter stain-----Diluted Carbol fuchsin 30 sec
Gram Positive
⚫ Have thick layer of
peptidoglycan

⚫ Violet/ purple

41
Gram Positive Cocci
Gram Positive Rods (Bacillus)
Gram Negative Bacteria
⚫ Thin layer of peptidoglycan

⚫ Red

44
Gram stain of urethral pus

45
 Shigella dysenteriae (gram –ve
bacilli)
 Ziehl-Neelsen staining
❖ Primary stain-----Fuming Carbol fuchsin 5 min

❖ Decolorizer--------20% H2SO4
❖ Counter stain-----Methylene blue 2-5 min

red, rod, beaded, slightly curved (AFB)

Mycobacterium tuberculosis (acid fast bacilli)
 Albert Staining

Corynebacterium diphtheriae
Types of culture media
❖ Based on their consistency
a) solid medium--- BA, CA,MCA, Nutrient agar
b) liquid medium – Trypticase soy broth, Nutrient broth
c) semi solid medium– TSI/KIA, MIU
❖ Special media
⚫ Enriched media- BA, CA
⚫ Selective media- MCA, L-J
⚫ Indicator media – MCA, BA, MIU
⚫ Transport media – Cary-Blair media, Amies media,
Alkaline peptone water
Blood agar Chocolate agar
Beta Hemolytic Streptococci
- secrete hemolysins that cause the complete lysis of RBC’s
Lactose Non-Lactose
Fermenter Fermenter

MacConkey Agar
Potassium Tellurite media LJ media
MIU
media
Urease medium
 Blood Cultures: Methods
1. Conventional method

2. Lytic-centrifugation method

3. Automated method

Liquid media
1. Conventional blood culture method

Media : Trypticase soya broth media /

Tryptone soya broth media

Content of Media: (broth +SPS+PABA)

Media Incubated for : 10 days – 2 weeks

SPS (sodium polyanethol sulfonate) ; SPS-antibotic neutralizer, PABA-enrichment

Blood culture bottles (Trypticase soya broth media )
2. Lytic-centrifugation blood culture method
3. Automated blood culture method

Media: Bactec Blood culture broth

Detect CO2
Types of Media:
1. Aerobic media (30 ml)
2. Anaerobic media (25 ml)

FAN BOTTLE
Automated Methods
 Antimicrobial
susceptibility testing
(AST)
 Types of AST
Conventionally performed phenotypic
method
• Disk diffusion Method: Kirby-Bauer’s disk
diffusion (DD) test, Stokes method
• Dilution tests:
a) Broth dilution method: Broth tube microdilution and broth microdilution
b) Agar dilution
• Diffusion and dilution method: Epsilometer or E-test
Automated AST
VITEK/ Phoenix/ Microscan (Principle: Broth microdilution)
Current methods for antimicrobial susceptibility testing and turnaround time (created with BioRender.com,
accessed on 27 February 2022. Reproduction of this figure requires permission from BioRender.com)
Kirby-Bauer’s Disk Diffusion Test
⚫ The method is based on placing different
antibiotic-impregnated disks on previously
inoculated agar with bacterial suspension.
⚫ Media: Mueller Hinton Agar (MHA)
⚫ The antibiotic diffuses radially outward through the
agar medium, producing an antibiotic concentration
gradient.
⚫ After the inhibition zones are established within 24
h of incubation at 35 ― 1◦C, the zone diameters of
each tested antibiotic are measured by the naked
eye or using an automated system.
Antibiotic sensitivity testing
Antibiotic susceptibility testing:

⚫Sensitive
⚫Intermediate sensitive
⚫Resistance
⚫ MIC
The lowest concentration (in μg/mL) of an
antibiotic that inhibits the growth of a
given strain of bacteria.

⚫ MBC
The minimum concentration of an
antimicrobial drug that is bactericidal. It is
determined by re-culturing (subculturing)
broth dilutions that inhibit growth of
bacteria (i.e., those at or above the MIC).
Antimicrobials

2
 Antibiotics
• Definitions: An antibiotic is a product
produced by microorganisms or a
similar substance produced by
chemical synthesis, which in low
concentration, kills or inhibits the
growth of microorganisms.
⚫Bactericidal drugs

⚫Bacteriostatic drugs

⚫Selective toxicity:
A drug should selectively kill or
prevent growth of a pathogen, but not
of host cells.
 Mechanism of action:
Antibiotics
⚫ Inhibition of cell wall synthesis
⚫ Inhibition of protein synthesis
⚫ Inhibition of nucleic acid synthesis
⚫ Alteration of cell membrane function
⚫ Others
CELL WALL SYNTHESIS INHIBITORS
β-Lactam Antibiotics
⚫ Penicillins (Source: Penicillium notatum)
⚫ Cephalosporins
⚫ Carbapenems
⚫ Monobactams

Other groups
⚫ vancomycin
⚫ Bacitracin
Classification of Penicillin
❖ Natural Penicillins
⚫ Benzylpenicillin (penicillin G) – I/M, I/V
Phenoxymethylpenicillin (penicillin V) -- Oral

❖ Semisynthetic Penicillins
Penicillinase-resistant penicillins
⚫ cloxacillin, flucloxacillin, methicillin
Aminopenicillins
⚫ amoxicillin, ampicillin
Anti pseudomonal penicillins
⚫ piperacillin, ticarcillin, carbenicillin, azlocillin
Penicillins:Mechanism of Action
Penicillin

Inhibit transpeptidase

Inhibit cross linking between pentapeptide

Inhibition of peptidoglycan synthesis of

bacterial cell wall

Bactericidal action
 Classification of Cephalosporins
Generation of cephalosporin antibiotics
Route
first second third fourth fifth

Oral Cephradin Cefaclor Cefixime - -

Cephalexin Cefuroxime
Cefadroxil

Inj Cephradin Cefuroxime Ceftriaxone Cefepime Ceftaroline,

Cephalexin Ceftazidime Cefpirome Ceftobiprole
Cefadroxil Cefotaxime

G+ve +++ ++ + ++ ++
G-ve +/- + +++ +++ +++
Carbapenems
Imipenem
Meropenem
Ertapenem

Monobactams
Aztreonam
 Vancomycin
M/A: Bind directly to the D-Ala—D-Ala
end of the peptidoglycan 🡪 Inhibits
bacterial cell wall synthesis
Uses:
•To treat MRSA
• Pseudomembranous colitis
 Inhibition of protein synthesis
⚫ Chloramphenicol,
Erythromycin, Clindamycin,
Linezolid (inhibit 50s)

⚫ Tetracyclines
Aminoglycosides (inhibit 30s)

Tetracyclines: Bacteriostatic
Complications of aminoglycosides:Ototoxicity,Nephrotoxicity,Neurotoxicity
⚫ Inhibition of Nucleic acid synthesis
❖ Sulfonamides, trimethoprim

❖ Quinolones, e.g. ciprofloxacin (inhibit DNA gyrase)

❖ Rifampicin (inhibit mRNA synthesis)
 Metronidazole
⚫ It act as an electron sink. By accepting
electrons, the drug deprives the organism of
required reducing power.

⚫ Damages DNA. When electrons are

acquired, the drug ring is cleaved and a toxic
intermediate is formed that damages DNA.
AWaRe Classification

2
 Chemoprophylaxis
⚫ antimicrobial agents used to prevent
diseases from occurring-chemoprophylaxis.
 Antimicrobial
resistance (AMR)

2
 Mechanism of Antibiotic
Resistance
⚫Genetic basis
* Chromosome-mediated
* Plasmid-mediated
* Transposon-mediated
⚫Non genetic basis
Genetic mechanism of AMR

2
Specific Mechanisms of Resistance
(Genetic basis)
Penicillins & Cephalosporins:
⚫ β-lactamases
⚫ Mutation in PBP
⚫ Poor permeability by mutation in porin proteins
⚫ Tolerance
 Vancomycin:
change in the peptide component of peptidoglycan
from D-alanyl-D-alanine, which is the normal
binding site for vancomycin, to D-alanine-D-lactate.

Aminoglycosides:
1. Mutation to ribosomal binding site
2. Decreased uptake of antibiotic
3. Enzymatic modification of the antibiotic.
 Tetracyclines
⚫ Reduction of the uptake of the drug or
⚫ Enhance its transport out of the cell.

Erythromycin
• Efflux pumps
• Methylation of a specific adenine on the 23S
• Decreases binding affinity of erythromycin
Sulfonamides
mutation of dihydropteroate synthetase

Trimethoprim
mutations of dihydrofolate reductase

Quinolones
mutations of bacterial DNA gyrase
Non genetic mechanism of AMR
Misuse and overuse of antimicrobials
Lack of access to clean water, sanitation and
hygiene (WASH) for both humans and animals
Poor IPC in health-care facilities and farms
Poor access to quality, affordable medicines,
vaccines and diagnostics
Unnecessary use of antibiotics in agriculture
Lack of awareness and knowledge
Lack of enforcement of legislation
(Source: WHO AMR Fact Sheet 2020; US-CDC) 2
Selection Pressure

2
Cross-resistance:
microbes resistant to a certain drug
may also be resistant to other drugs
that share a mechanism of action.
(e.g., different aminoglycosides,
macrolides, and lincomycins)
Impact
of
AMR:

https://antimicrobialsworkinggroup.
org/antimicrobial-resistance/
A 10 point
plan to
combat
AMR

Source: Review
on Antimicrobial
Resistance 2014
Place your logo in
white here

World AMR Awareness Week

(WAAW)

18-24 November, 2023

2
MDR, XDR, PDR
 MDR, XDR, PDR

TABLE 1. Staphylococcus aureus; antimicrobial categories and agents used to define MDR, XDR and PDR (worksheet for categorizing isolates)
 MDR, XDR, PDR
⚫ MDR: Non-susceptibility to at least one
agent in three or more antimicrobial
categories.

⚫ XDR: Non-susceptibility to at least one

agent in all but two or fewer antimicrobial
categories (i.e. bacterial isolates remain
susceptible to only one or two categories).

⚫ PDR: Non-susceptibility to all agents in

all antimicrobial categories.
Based on expert consensus, CDC and ECDC gave us a widely used set of ideas in Magiorakos et al. (CMI 2012)
 Multidrug-resistant
tuberculosis (MDR)
• Resistance to at least rifampicin
and isoniazid, with or without other
drug resistance.

Ref: Davidson’s Medicine 24th ed, 2022

 Extensively drug-resistant
tuberculosis (XDR)
•Resistance to at least rifampicin and
isoniazid, in addition to any
quinolone and at least one injectable
second-line agent
Ref: Davidson’s Medicine 24th ed, 2022
 Sterilization
The process of destruction or
removal of all microorganisms
including spores.

⚫ Sterilant
 Disinfection
The procedure which inactivates all
recognized pathogenic
microorganisms but not necessarily
all microbial life form, i.e- highly
resistant bacterial endospores.

⚫ Disinfectant
 Antisepsis
The procedure which inhibits or
destroys microorganisms in living
tissue used in skin and mucous
membrane.

⚫ Antiseptic
 Sterilant Disinfectant Antiseptic

Procedure Sterilization Disinfection Antisepsis

Act upon Non living Non living/ Living/

Object inanimate animate
surface surface
⚫Bactericide:
An agent that kills bacteria.
Most do not kill Endospores.

⚫ Sporicide:
An agent that kills spores.
 Physical Methods
⚫ Heat:
1. Dry heat: Red heat, Flaming, Hot Air Oven
2. Moist heat:
At 100°C– Steaming (Tyndallization)
> 100°C – Autoclaving
⚫ Radiation: Ionizing radiation, UV radiation
⚫ Filtration: Berkefeld filter, Asbestos filter etc.
 Uses of Hot Air Oven:
⚫ Glasswares - Syringe, petridishes,
Test tube, Flask, Glass pipette.

⚫ Swab stick.

⚫ Liquid paraffin, fats, grease,

Ointment, Oils, Waxes, Powders.
 Tyndallization
⚫ Fractional / Intermittent sterilization
⚫ Steaming at 100°C for 20 min
followed by incubation at 37°C
overnight for 3 successive days
⚫ Use: Heat labile media containing
sugar, egg, serum, gelatin etc.
 Autoclaving
⚫ The process of sterilization by steam
under high pressure above 100°C
⚫ Principle:
15 lb/inch2
121°C
15-20 min
⚫ Condensation of heat
Autoclaving
Use:
⚫ Culture media
⚫ Medical & surgical instruments
except blunt instruments
⚫ Suture materials except catgut
⚫ Dressing
⚫ Gown
Monitoring of autoclaves

⚫ Biological Indicator:
B. stearothermophilus

⚫ Chemical Indicator:
⚫ Autoclave tape
⚫ Browne’s tube
it consists of heat sensitive
chemical that changes color at
the right temperature and
exposure time.
 Pasteurisation
⚫ Used to kill pathogens in milk
⚫ Not sterilization
⚫ Method Temp Time
o
Holding (Batch) 63 C 30 min
o
Flash 72 C 15 sec
Ultrahigh-temperat 134ºC 1 sec
ure (UHT)
Milk borne pathogens:
⚫ Mycobacterium bovis
⚫ Salmonella spp
⚫ Escherichia coli
⚫ Brucella spp
⚫ Listeria spp
⚫ Coxiella burnetti
⚫ Polio virus
Use of Ɣ-Ray
⚫ Disposable syringe, Needle
⚫ Transfusion equipments
⚫ Catgut
⚫ Nylon sutures
⚫ Gloves
⚫ Adhesive dressing
⚫ Bone & Tissue graft
 Use of UV-Ray
⚫ Air disinfection in biosafety cabinet
⚫ Operation Theatre (OT)
⚫ Microbiological Lab
⚫ Hospitals, Schools
⚫ Pharmaceutical industries
 Chemical Methods
⚫ Disruption of cell membrane:
- Detergents, Alcohol
- Phenol (Cresol/Lysol, Chlorhexidine)
⚫ Protein denaturation:
- Benzoic acid, L. acid, Citric acid
⚫ Modification of Protein & NA:
- Ethyline oxide, Formaldehyde
- Gluteraldehyde, Halogens (Cl2, I2)
- Hypochorite, H2O2, Heavy metal, Dye
 Alcohol
⚫ 70% Ethyl alcohol
⚫ 70-90% Isopropyl alcohol
⚫ Use:
- Disinfection of clinical thermometer
- Disinfection of the skin: Venupuncture
 Phenol
⚫Cresol
Use: Sharp instruments
Floor of ward, OT
⚫Chlorhexidine
Use: Skin antisepsis

Benzoic acid: Weak antiseptic

Use of Ethylene oxide gas
⚫Heart-Lung machine
⚫Plastic & rubber goods-
Ambu bag, Catheter
⚫Polyethylene tube, DPS
⚫Prosthetic heart valve
⚫Medical equipments
Use of Formaldehyde (40%)
⚫Rooms
⚫Beddings
⚫Clothings
⚫Hides & wools
 Gluteraldehyde
⚫ 2% Gluteraldehyde for 20 min
--- High level disinfection
⚫ 2% Gluteraldehyde for 3-12 hrs
--- Chemical sterilization
Use of Gluteraldehyde
⚫Endoscopic instruments
⚫Transducers
⚫Dialysis system
⚫Anaesthesia euipments
⚫Ventlation system
Use of Hypochlorite (Bleach)
⚫ Dialysis system
⚫ Hydrotheraphy tanks
⚫ Dental appliances
⚫ CPR equipments
⚫Toilets
Heavy metals
⚫1% AgNO3
Use: Prophylaxis of Ophthalmia
neonatorum

⚫Silver sulphadiazine
Use: Burn patients
Common methods of Sterilization
⚫Autoclave
⚫Hot air oven
⚫Ɣ radiation
⚫Gluteraldehyde
⚫Ethylene oxide
⚫Tyndallization
 Normal human Microbiota
(Normal flora / Commensals)

Various bacteria and fungi that are

permanent residents of certain body
sites, especially the skin, oropharynx,
colon, and vagina.

149
Location Important Less Important Organisms
Organisms
Skin Staph Staph aureus, diphtheroids
epidermidis various streptococci,
Pseudo aeruginosa,
anaerobes
(Propionibacterium),
yeasts (C albicans)

Nose Staph aureus S. epidermidis,

Coryne (diphtheroids),
various strepto
150
Location Important Organisms Less Important
Organisms
Mouth Viridans Various strepto,
streptococcus Eikenella corrodens

Dental Streptococcus Prevotella intermedia,

plaque mutans Porphyromonas
gingivalis

Gingival Various anaerobes,

crevices e.g., Bacteroides,
Fusobacterium,
streptococci,
Actinomyces
151
Throat Viridans Various streptococci (Strepto
streptococcus pyogenes & Strepto
pneumoniae),
Neisseria species,
Haemophilus influenzae,
S. Epidermidis

Colon Bacteroides Bifidobacterium,

fragilis, Eubacterium,
E coli Fusobacterium,
Lactobacillus, various aerobic
gram-negative rods,
Enterococcus faecalis and
other streptococci, Clostridium

152
Vagina Lactobacillus, Various streptococci,
E. Coli, various gram-negative
group B rods
streptococcus B. fragilis,
Corynebacterium
(diphtheroids),
C. albicans
Urethra S. epidermidis,
Corynebacterium
(diphtheroids), various
streptococci, various
gram-negative rods,
e.g., E. coli

153
Benefits of Normal Flora
⚫ Colonization resistance
⚫ Mucosal maturation
⚫ Stimulate Immune System
⚫ Synthesis of vitamins (B, K)
by E. coli & Bacteroids
⚫ Digestion
⚫ Probiotics

154
 Colonization resistance
⚫ The ability of members of the normal
flora to limit the growth of pathogens.

⚫ For example, antibiotics can reduce the

normal colonic flora that allows Clostridium
difficile, which is resistant to the antibiotics, to
overgrow and cause pseudomembranous
colitis.

155
Probiotics:
⚫ Living organisms administrated orally
to promote health
⚫ Mechanism speculative: competition with other
bacteria; stimulation of nonspecific immunity
⚫ Species specific: adherence & growth (tropism)
⚫ Example: Lactobacillus rhamnosus
Saccharomyces boulardii

156
Harmful effects of Normal Flora
⚫ Opportunistic pathogens

⚫ Production of carcinogen

157
 Opportunistic flora
⚫ Some normal flora become opportunistic
pathogens
⚫ (Staphylococcus aureus, Streptococcus mutans,
Enterococcus faecalis, Streptococcus
pneumoniae, Pseudomonas aeruginosa, etc.)
⚫ Breach of skin/mucosal barrier: trauma, surgery,
burns
⚫ Bacterium at one site may be commensal, but
might be pathogenic at another site

158
⚫ Production of carcinogen
Some normal flora may modify, through their
enzymes, some chemicals in our diets into
carcinogens.
e.g. Artificial sweeteners may be enzymatically
modified into bladder carcinogens

159
Pathogenesis of
bacterial diseases

160
Typical Stages of an Infectious Disease:

⚫ Incubation period which is the time between the acquisition of the organism
(or toxin) and the beginning of symptoms (this time varies
from hours to days to weeks, depending on the organism).

⚫ Prodrome period during which nonspecific symptoms such as fever, malaise,

and loss of appetite occur.

⚫ Specific-illness period overt characteristic s/s of the disease occur.

⚫ Recovery / Convalescence period

161
 Stages of Bacterial Pathogenesis:
1. Transmission from an external source into the portal of entry.
2. Evasion of primary host defenses such as skin or stomach
acid.
3. Adherence to mucous membranes, usually by bacterial pili.
4. Colonization by growth of the bacteria at the site of
adherence.
5. Disease symptoms caused by toxin production or invasion
accompanied by inflammation.
6. Host responses, both nonspecific and specific (immunity),
during steps 3, 4 & 5.
7. Progression or resolution of the disease.

162
Important Modes of Transmission:

Human Non
human

Human

163
 Human to Human transmission:

⚫ Horizontal transmission: person-to-person

⚫ Vertical transmission : mother to offspring

164
 Route of entry in Horizontal transmission:

⚫ Respiratory route

⚫ Gastrointestinal route

⚫ Skin

⚫ Genital route

⚫ Parenteral route

165
 Vertical Transmission:
⚫ Transplacental: Treponema pallidum, Listeria
monocytogenes, Toxoplasma gondii.

⚫ Within birth canal/at the time of birth:

Streptococcus agalactiae(group B streptococci),
Escherichia coli, Chlamydia trachomatis,
Neisseria gonorrhoeae.

⚫ Breast milk: Staphylococcus aureus

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 Transmission of Waterborne Diseases:

1. Ingestion of drinking water

⚫ Salmonella
⚫ Shigella
⚫ Campylobacter jejuni
2. Ingestion of water while swimming
⚫ Leptospira interrogans

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Bacterial Diseases Transmitted by Foods:
I. Diarrheal diseases
⚫ Staphylococcus aureus
⚫ Bacillus cereus
⚫ Clostridium perfringens
⚫ Escherichia coli
⚫ Salmonella enteritidis
⚫ Shigella species
⚫ Vibrio cholerae

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Bacterial Diseases Transmitted by Foods:
II. Nondiarrheal diseases
⚫ Clostridium botulinum
⚫ Listeria monocytogenes
⚫ Mycobacterium bovis
⚫ Brucella species
⚫ Francisella tularensis

169
Bacterial Zoonotic Diseases:
(Animal source to human)

⚫ Bacillus anthracis --- Anthrax

⚫ Listeria monocytogenes --- Neonatal sepsis
⚫ Bartonella henselae --- Cat-scratch disease
⚫ Brucella species --- Brucellosis
⚫ Campylobacter jejuni --- Diarrhoea
⚫ Escherichia coli O157:H7 -- Hemorrhagic colitis
⚫ Yersinia pestis --- Sepsis
⚫ Mycobacterium bovis --- Intestinal tuberculosis

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 Bacterial Virulence factor:
⚫ Polysaccharide capsule:
Streptococcus pneumoniae, Neisseria
meningitides, Haemophilus
influenzae, Klebsiella pneumoniae
⚫ Polypeptide capsule: Bacillus anthracis
⚫ M protein: Streptococcus pyogenes
⚫ Protein A: Staphylococcus aureus

171
 Bacterial Virulence factor:
⚫ Pili: Neisseria gonorrhoeae, E. coli
⚫ Glycocalyx: Staphylococcus epidermidis
certain viridans streptococci
⚫ Adhesins (Biofilm production)- Pseudomonas
⚫ Curli: E. coli, Salmonella spp.

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Several enzymes for invasiveness:

⚫ Collagenase, hyaluronidase:
Strep. pyogenes
⚫ Coagulase: Staph. aureus
⚫ IgA protease: N.gonorrhoeae,
Haemophilus influenzae,
Streptococcus pneumoniae
⚫ Streptokinase, Streptodornase:
Streptococcus pyogenes

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 Bacterial Toxins:

⚫Exotoxins Gram-positive bacteria

Gram negative bacteria

⚫Endotoxins Gram-negative bacteria

174
 Property Exotoxin Endotoxin
Source Certain species of Cell wall of
gram-positive and gram-negative
gram-negative bacteria bacteria
Secreted Yes No
from cell
Chemistry Polypeptide LPS

Location of Plasmid or Bacterial

genes bacteriophage chromosome
Toxicity High (fatal dose on the Low (fatal dose
order of 1 µg) on the order of
100 µg)
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 Property Exotoxin Endotoxin
Mode of Various modes Includes TNF and
action interleukin-1
Antigenicity Induces high-titer Poorly antigenic
antibodies called
antitoxins
Vaccines Toxoids used as No toxoids formed
vaccines and no vaccine
available
Heat Destroyed rapidly at Stable at 100°C for
stability 60°C (except 1 hour
staphylococcal enterotoxin)
Typical Tetanus, botulism, Meningococcemia,
diseases diphtheria sepsis
176
Important Bacterial Exotoxins:
⚫ Diphtheria toxin:
Corynebacterium diphtheriae
⚫ Tetanus toxin (tetanospasmin):
Clostridium tetani
⚫ Botulinum toxin: Clostridium botulinum
⚫ TSST, Enterotoxin, Exfoliatin:
Staph. aureus

177
 Important Bacterial Exotoxins:
⚫ Erythrogenic toxin: Streptococcus
pyogenes
⚫ Enterotoxin (choleragen):
Vibrio cholerae
⚫ Enterotoxin (LT, ST), Shiga toxin
(verotoxin): E. coli

178
Important exotoxins having A–B subunit
⚫ Diphtheria toxin
⚫ Tetanus toxin
⚫ Botulinum toxin
⚫ Cholera toxin
⚫ Enterotoxin of ETEC

179
ADP-ribosylation
⚫ Diphtheria toxin
⚫ Cholera toxin
⚫ Enterotoxin of E. coli
⚫ Pertussis toxin
⚫ Pseudomonas exotoxin A

180
Mechanism of Action of Diphtheria toxin:

Diphtheria toxin (A-B)

EF-2 + NAD EF-2- ADP-ribose + Nicotinamide

Inactivation of EF-2

Inhibition of protein synthesis

181
Mechanism of Cholera toxin :
Subunit B attaches to the ganglioside at the
brush border of epi. cells of small intestine
Facilitates the entry of subunit A Subunit A
dissociates and penetrate cell membrane
addition of ADP-ribose (ADP-R) to the GS regulatory
protein activates adenyle cyclase cAMP

1.prolonged hypersecretion of water

2.Increase Na dependent Cl secretion
3.inhibits the reabsorption of Na and Cl
4.gut lumen is distended with fluid
5.hypermotility and diarrhea occur
182
Mechanism of LT of ETEC
Subunit B

attaches to the ganlioside at the brush border

of epi. cell of small intestine Facilitates
the entry of subunit A ADP-ribosylation
of Gs protein activates adenyle cyclase
increases cAMP prolonged
hypersecretion of water &chloride and inhibits
the reabsorption of sodium gut lumen is
distended with fluid, hypermotility and
diarrhea occur.
183
 Endotoxins
⚫ Endotoxins are integral parts of the cell
walls of both gram-negative rods and
cocci
⚫ LPS in nature

184
185
186
 Bacterial Infections associated with
Cancer / Oncogenic bacteria

⚫Helicobacter pylori
Gastric carcinoma and gastric
mucosal-associated lymphoid tissue
(MALT) lymphoma
⚫Campylobacter jejuni
MALT lymphoma of the small intestine
(alpha-chain disease)

187
Koch's postulates (Robert Koch,1877):
1. The organism must be isolated from every
patient with the disease.
2. The organism must be isolated free from all
other organisms and grown in pure culture in
vitro.
3. The pure organism must cause the disease in a
healthy, susceptible animal.
4. The organism must be recovered from the
inoculated animal.

188
Limitation of Koch’s postulates
⚫ Non-culturable microbial diseases-
Syphilis, Leprosy
⚫ Microorganisms with out having any
known susceptible animals -
N. gonorrhoea
⚫ Genetic diseases- Thalassaemia
⚫ Auto-immune diseases- SLE

189