Residency Preparation

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 Mentor

Dr. Fahim Uddin Ahmed

MBBS, MD (Microbiology)

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 Today’s Topic

Topic: Basic Bacteriology

FCPS, Residency (MD/MS), Diploma, MRCP পরীক্ষার ��িত িনেত

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Important topics of General bacteriology
• Structure of bacteria

• Bacterial growth

• Classification of Medically important bacteria

• Human Microbiome

• Pathogenesis and host defenses

• Antibacterial drugs and resistance

• Bacteria vaccine
Important topics-Structure of bacterial cell
• Important characteristics or difference of eukaryotic and prokaryotic cell-Selective
toxicity

• Essential and non-essential structure of bacterial cell with their functions.

• Composition and function of cell wall of gram positive and gram negative bacteria

• What are the cell wall acting antibiotics and how they became resistance

• Function and clinical significance of Periplasmic space, capsule, pilli , flagella –

Example of these structural presence among bacteria.

• Plasmid-clinical significance

• Spore- structure, significance, example.

Important topics- Bacterial growth, classification, Pathogenesis

• Importance of growth curve

• Example of Gram positive, Gram negative, non-gram stain bacteria

• Example of normal flora in different anatomical location

• Example of exotoxin, endotoxin, superantigen with their

characteristics

• Example of fastidious bacteria

• Example of facultative intracellular bacteia

Important topics-Sterilization, Antibiotics, Vaccine

• Methods of sterilization

• Procedure of different material sterilization

• High level, intermediate level and low level disinfectant

• Different mechanism of antibiotic action

• Example of antibiotics with different mechanism of action

• Mechanism of drug resistance

• Example of Bacterial vaccine

Eukaryotes Vs Prokaryotes
Structure of bacterial cells
Structure of bacterial cells
Cell wall of Bacteria
 Cell wall of Bacteria

(A) A gram-positive bacterium has a thick peptidoglycan layer that contains teichoic and lipoteichoic acids. (B) A gram-negative bacterium has a thin
peptidoglycan layer and an outer membrane that contains lipopolysaccharide, phospholipids, and proteins. The periplasmic space between the cytoplasmic and
outer membranes contains transport, degradative, and cell wall synthetic proteins. (C) Mycobacterial cell walls confer acid-fast staining to the bacteria. They
have a complex structure with a lipid rich waxy outer layer of mycolic acids with porins that permeate that layer.
Comparison of Cell Walls of Gram-Positive
and Gram-Negative Bacteria
Cell Walls of Gram-Negative Bacteria
Bacterial cell wall: Peptidoglycan (murein
and mucopeptide)
Clinical importance of peptidoglycan layer : Action of antibiotics

• Several of drugs, such as penicillins, cephalosporins, and vancomycin,

inhibit the synthesis of peptidoglycan by inhibiting the transpeptidase

that makes the cross-links between the two adjacent tetrapeptides.

Clinical importance of peptidoglycan layer:
Action of antibiotics
Protoplast, spheroplast, L-form
 Cell wall component: Lipopolysaccharide
• The lipopolysaccharide (LPS) of the outer membrane of the cell wall of
gram-negative bacteria is endotoxin.

• It is responsible for many of the features of disease, such as fever and

shock (especially hypotension).
Structure of LPS
• The LPS is composed of three distinct units :

• (1) A phospholipid called lipid A, which is responsible for the toxic effects.

• (2)A core polysaccharide of five sugars linked through

ketodeoxyoctulonate (KDO) to lipid A.

• (3) An outer polysaccharide consisting of up to 25 repeating units of three

to five sugars.
Structure of LPS
Structure of LPS
• This outer polymer is the important somatic, or O, antigen of several
gram negative bacteria that is used to identify certain organisms in
the clinical laboratory.

• Some bacteria, notably members of the genus Neisseria, have an

outer lipooligosaccharide (LOS) containing very few repeating units
of sugars.
Cell wall component: Teichoic Acid
• Teichoic acids are fibers located in the outer layer of the gram-
positive cell wall.
 Importance of teichoic acid
• The medical importance of teichoic acids lies in their ability to induce

inflammation and septic shock when caused by certain gram-positive

bacteria; that is, they activate the same pathways as does endotoxin

(LPS) in gram-negative bacteria.

Difference between gram positive vs gram
negative bacteria
 Previous year question
• Following are essential components of Bacterial structure

A) Cytoplasmic Membrane

B) Plasmid

C) Nucleoid

D) Mesosome

E) Flagella

T F T T F
Previous year question
• Cell wall of gram positive bacteria contains

A) Peptidoglycan

B) Periplasmic space

C) Lipopolysaccharide

D) Teichoic acid

E) Porin protein

T F F T F
Gram stain procedure
Medically Important Bacteria That Cannot Be Seen
in the Gram Stain
Previous year question
• Bacteria that are not visualized microscopically by gram stain includes

A) Borrelia

B) Mycobacteria

C) Yersinia

D) Chlamydia

E) Acinetobacter

T T F T F
Cytoplasmic Membrane
Cytoplasmic Membrane

• Composed of a phospholipid bilayer similar in microscopic appearance

to that in eukaryotic cells.

• They are chemically similar, but eukaryotic membranes contain sterols ,

whereas prokaryotes generally do not EXCEPT Mycoplasma.
Cytoplasmic Membrane

• The membrane has four important functions:

• (1) active transport of molecules into the cell,

• (2) energy generation by oxidative phosphorylation,

• (3) synthesis of precursors of the cell wall, and

• (4) secretion of enzymes and toxins.

Cytoplasm
Cytoplasm (Ribosomes)

• Bacterial ribosomes are the site of protein synthesis as in eukaryotic cells,

• But they differ from eukaryotic ribosomes in size and chemical

composition
Cytoplasm (Granules)

• Serve as storage areas for nutrients and stain characteristically with

certain dyes.

• For example, volutin is a reserve of high energy stored in the form of

polymerized metaphosphate. It appears as a “metachromatic” granule
found in Cornebacterium diptheriae.
Cytoplasm (Granules)
Nucleoid

Eukaryotic cell Bacterial cell

 Nucleoid

• Bacterial nucleoid contains no nuclear membrane, no nucleolus, no

mitotic spindle, and no histones and no introns
Plasmids
• Plasmids are extra chromosomal, double-stranded, circular DNA molecules
that are capable of replicating independently of the bacterial chromosome.

• Although plasmids are usually extra chromosomal, they can be integrated

into the bacterial chromosome.
 Types of Plasmids

• Plasmids occur in both gram-positive and gram negative bacteria, and

several different types of plasmids can exist in one cell:

• Transmissible plasmids can be transferred from cell to cell by conjugation

• Non-transmissible plasmids
Transmissible Plasmids
Function of Plasmids
• Plasmids carry the genes for the following functions and structures of medical
importance:

 (1) Antibiotic resistance, which is mediated by a variety of enzymes,

(2) Exotoxins,

(3) Pili (fimbriae),

(4) Resistance to heavy metals

(5) Resistance to ultraviolet light,

 (6) Bacteriocins
Previous year question
• Plasmid

A) Is chromosomal DNA

B) Contain linear DNA

C) Replicate independently of bacterial chromosome

D) Can be integrated into the bacterial chromosome

E) Present only in gram negative bacteria

F F T T F
Transposons/Jumping gene
• Transposons are pieces of DNA that move readily from one site to
another either within or between the DNAs of bacteria, plasmids, and
bacteriophages.

• Transposons can code for drug-resistant enzymes, toxins, or a variety of

metabolic enzymes and can either cause mutations in the gene into which
they insert or alter the expression of nearby genes
Transposons/Jumping gene
Structures Outside the Cell Wall- Capsule
• It is typically composed of polysaccharide.

• The sugar components of the polysaccharide vary from one species of

bacteria to another and frequently determine the serologic type
(serotype) within a species.

• For example, there are 91 different serotypes of Streptococcus

pneumoniae.
Capsule
• The capsule is important for four reasons:

• 1) It is a determinant of virulence of many bacteria since it limits the

ability of phagocytes to engulf the bacteria.

• 2) Specific identification of an organism can be made by using antiserum

against the capsular polysaccharide. (Quellung reaction)
Capsule
• 3) Capsular polysaccharides are used as the antigens in certain vaccines
because they are capable of eliciting protective antibdies.

• 4) The capsule may play a role in the adherence of bacteria to human

tissues, which is an important initial step in causing infection.
Capsule- Quellung reaction
Example of encapsulated bacteria
Most important encapsulated bacteria and fungi (YES Some Nasty Killers Have Pretty Big Capsules)

• Yersinia pestis

• Escherichia coli-meningeal strains only

• Salmonella typhi

• Streptococcus pneumoniae and Streptococcus pyogenes

• Neisseria meningitidis

• Klebsiella pneumoniae

• Haemophilus influenzae type B (B polysaccharide)

• Pseudomonas aeruginosa

• Bordetella pertussis and Bacillus anthracis (contains poly D-glutamate capsule)

• Cryptococcus neoformans- only encapsulated fungal pathogen

Previous year question
• Organism possess capsule are

A) Haemophillus influenza

B) Cryptosporidium spp.

C) Streptococcus pneumoniae

D) Staphylococcus saprophyticus

E) Klebsiella pneumoniae

T F T F T
 Flagella

• Flagella are long, whiplike appendages that move the bacteria toward

nutrients and other attractants, a process called chemotaxis.

• Flagella is composed of many subunits of a single protein, flagellin.

Importance of flagella
• Flagella are medically important for two reasons:

• (1) Some species of motile bacteria (e.g., E. coli and Proteus species) are
common causes of urinary tract infections.

• (2) Some species of bacteria (e.g., Salmonella species) are identified in the
clinical laboratory by the use of specific antibodies against flagellar proteins
Pili (Fimbriae)
• Pili are composed of subunits of pilin. They are found mainly on gram-
negative organisms.

• Two classes can be distinguished

Ordinary pili

 Sex pili
Pili (Fimbriae)
Importance of pili
• Pili have two important roles:

• (1) They mediate the attachment of bacteria to specific receptors on the

human cell surface.

• (2) A specialized kind of pilus, the sex pilus, forms the attachment between
the male (donor) and the female (recipient) bacteria during conjugation
Previous year question
• Bacterial pilli are involved in

A) Adherence to host cells

B) Transfer of genetic material

C) Resistance to heat

D) Endotoxin activity

E) Motility

T T F F F
Glycocalyx (Slime Layer)
• The glycocalyx is a polysaccharide coating that is secreted by many bacteria. It
covers surfaces like a film and allows the bacteria to adhere firmly to various
structures (e.g., skin, heart valves, prosthetic joints, and catheters).

• The glycocalyx is an important component of biofilms.

• Psudomonas aeruginosa, Staphylocoocus epidermidis, and viridans

streptococci are medically important bacteria producing biofilm.
Biofilms
 Bacterial Spores/Endospore
• These highly resistant structures are formed in response to adverse
conditions by two genera of medically important gram-positive rods:

• The genus Bacillus, which includes the agent of anthrax, and

• The genus Clostridium, which includes the agents of tetanus and

botulism
Important Features of Spores and Their Medical
Implications
Previous year question
• Bacterial spores are

A) Antibiotic sensitive

B) Thermostable

C) Reproductive stage in bacterial multiplication

D) Easily destroyed by gamma radiation

E) Sensitive to antiseptics

F T F T F
Previous year question
Endospore formation is seen in
• Clostridium diflicile
• Bacillus subtilis
• Brucella melitensis
• Coxiella burnetti
• Bacteroides fragilis

Answer: TTFFF
Bacterial growth curve
Importance of different phases of growth cycle
lag phase :

Membrane acting antibiotics , soap and detergent act better in this phase

log (logarithmic) phase :

Cell wall inhibiting drug act on this phase

Stationary phase :

Release of exotoxin and spore formation starts, Gram variability occurs.

Death phase:

Sporulation, Development of L-form occurs in this stage

Obligate intracellular growth
• Most bacterial pathogens of humans are capable of growing on artificial
media in the clinical laboratory

• However, certain bacterial pathogens of humans, notably Chlamydia and

Rickettsia and Ehrlichia and Anaplasma can only grow within living cells
and are referred to as obligate intracellular pathogens.
Facultative intracellular bacteria
• Facultative intracellular bacteria are capable of living and reproducing in or outside of host cells. Example:

 Bartonella henselae

 Francisella tularensis

 Listeria monocytogenes

 Salmonella Typhi

 Brucella

 Legionella

 Mycobacterium

 Nocardia

 Neisseria

 Yersinia
Previous year question
• The obligate intracellular bacteria are

A) Chlamydia

B) Rickettsia

C) Legionella

D) Mycobacterium

E) Mycoplasma

T T F F F
Previous year question
• Facultative intracellular organisms are

A) Rickettsia

B) Legionella

C) Chlamydia

D) Brucella

E) Yersinia

F T F T T
 Previous year question
Facultive intracellular microorganisms are
• Salmonella paratyphi
• Bacillus cereus
• Mycobacterium bovis
• Vibrio cholerae
• E.coli

T F T F F

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 Aerobic & Anarobic Growth

• Because the use of oxygen generates two toxic molecules, hydrogen

peroxide (H2O2) and the free radical superoxide (O2 – ), bacteria require
two enzymes to detoxify these molecules when oxygen is used
 Aerobic & Anarobic Growth
• The first is superoxide dismutase, which catalyzes the following reaction:

2O2 – + 2H+ → H2O2 + O2

• The second is catalase, which catalyzes the following reaction:

2H2O2 → 2H2O + O2
Culture media
Previous year question
• Anaerobic bacteria cannot grow in presence of oxygen because

A) Lack of metabolic activity

B) Lack of superoxide dismutase

C) Lack of catalase

D) lack of oxidase

E) High content of oxygen

F T T F F
Classification of Medically Important Bacteria
Previous year question
• Following are gram negative bacteria

a) Pseudomonas aeruginosa

b) Treponema pallidum

c) Yersinia enterocolitica

d) Helicobacter pylori

e) Vibrio cholera

T F T T T
Human microbiome
• Members of the microbiome are considered permanent
residents of the associated body sites, such as the skin,
oropharynx, colon, and vagina

• These microbes are often referred to as commensals.

Anatomical location of normal flora
Anatomical location of normal flora
Summary of the Members of Normal Flora
and their Anatomic Locations
Medically Important Members of the Normal Flora
Previous year question
• Normal vagianal flora includes

A) Anaerobic streptococci

B) E.coli

C) Staphylococcus aureus

D) Lactoibacillus

E) Streptococcus pyogens

T T F T F
Infection Control Precautions and Practices
 Sterilization, Disinfection
• STERILIZATION:

Is defined as a process by means of which an article, surface or medium is

made free from all living microorganisms-including spores.

• Disinfection:

Is a process of destruction of vegetative forms of pathogenic infection but not

necessarily resistant to spores.
Methods of sterilization
• A) Physical methods
I) Sunlight
II) Drying: Spores remain uneffected
III)Heat:
 Dry heat
-Red heat: Metallic objects
-Flaming : Glass slide
-Incineration : Soiled dressing, bedding, pathological materials.
-Hot air oven : Glassware like Petri dishes, test tubes
 Methods of sterilization
Moist heat

1. At temperature below 100 C

A) Vaccine bath:

B) Pasturization

C) Inspissator

2. At temperature of 100 C

A) Boiling

B) Stem at atmospheric pressure

C) Tyndalization

3. Steam under pressure at temperature above 100 C

A) Autoclave
Methods of sterilization
IV) Filtration

V) Radiation

VI) Ultrasonic and sound vibrations

 Methods of sterilization
B) Chemical methods:

• I) Alcohols

• II) Aldehydes

• III) Phenolic compounds

• IV) Quaternary ammonium compounds

• V) Halogens

• VI) Oxidizing agents

• VII) Salts of heavy metals

• VII) Dyes

• VIII) Gas vapour sterilization

Disinfection
• High-level disinfectants are used for items involved with invasive
procedures that cannot withstand sterilization procedures (e.g., certain
types of endoscopes and surgical instruments with plastic or other
components that cannot be autoclaved).

• Disinfection of these and other items is most effective if cleaning the

surface to remove organic matter precedes treatment.

• Examples of high-level disinfectants include treatment with moist heat and

use of liquids such as glutaraldehyde, hydrogen peroxide, peracetic acid,
and chlorine compounds.
 Disinfection
• Intermediate-level disinfectants (i.e., alcohols, iodophor compounds,
phenolic compounds) are used to clean surfaces or instruments on which
contamination with bacterial spores and other highly resilient organisms is
unlikely.

• These have been referred to as semicritical instruments and devices and

include flexible fiberoptic endoscopes, laryngoscopes, vaginal specula,
anesthesia breathing circuits and other items.
 Disinfection
• Low-level disinfectants (i.e., quaternary ammonium compounds) are
used to treat noncritical instruments and devices, such as blood
pressure cuffs, electrocardiogram electrodes, and stethoscopes.

• Although these items come into contact with patients, they do not
penetrate through mucosal surfaces or into sterile tissues.
Clinical Use of Disinfection and Sterilization
Sterilization of surgical instruments and heat-
sensitive materials, such as endoscopes
• Ethylene Oxide (EtO) Gas Sterilization:

• Low-Temperature Hydrogen Peroxide Gas Plasma:

• Peracetic Acid Sterilization:

• Ozone Sterilization:

• Ultraviolet (UV) Light Sterilization:

Each method has its specific applications, advantages, and limitations. The
choice of method depends on the type of equipment, its material, the nature of
the medical procedure, and the institutional protocols.
Germicidal Properties of Disinfectants and Antiseptic
Agents
Previous year question
• Disposable syringes are sterilized by

A) Ethylene oxide

B) Formaldehyde

C) Ionizing radiation

D) UV radiation

E) Hexachloride

T F T F F
Previous year question
• Endoscopes are sterilized by
A) Plasma sterilization
B) Formaldehyde solution
C) Hydrogen peroxide
D) Boiling
E) 2% glutaradehyde solution

F T T F T
Previous year question
• Following are high level disinfectants

A) Plasma sterilization

B) Formaldehyde solution

C) Hydrogen peroxide

D) Boiling

E) 2% glutaraldehyde solution

F T T F T
Previous year question
• Physical methods of sterilization include

A) Radiation

B) Sterilization by ethylene oxide

C) Filtration

D) Boiling

E) Pasturization

T F T T T
Antimicrobial drug stewardship
• The basic principles of good stewardship are threefold:

• (1) reduce inappropriate use of antibiotics,

• (2) encourage targeted treatment with narrow-spectrum drugs, and

• (3) limit adverse effects

Basic Principles of Antimicrobial Drug Stewardship
Principles of antimicrobial therapy

• The most important concept underlying antimicrobial therapy is selective

toxicity.

• Broad-spectrum antibiotics (tetracyclines )

• Narrow-spectrum (vancomycin)
Bactericidal & bacteriostatic activity

• A bactericidal drug kills bacteria, whereas a bacteriostatic drug inhibits their

growth but does not kill them.

• The salient features of the behavior of bacteriostatic drugs are that

• (1) the bacteria can grow again when the drug is withdrawn, and

• (2) host defense mechanisms, such as phagocytosis, are required to kill the
bacteria.
Mechanism of Action of Important Antibacterial Drugs
Model of typical bacterial cell showing sites of action
of important antibacterial drugs
Model of typical bacterial cell showing sites of action
of important antibacterial drugs
Previous year question
Antibiotics act by inhibiting cell wall synthesis are -
• Gentamicin
• Vancomycin
• Meropenem
• Streptomycin
• Cefixim
Answer: FTTFT
Previous year question
Antimicrobial agents that act by inhibition of protein synthesis are

a) Streptogramins

b) Nalidixic acid

c) Vancomycin

d) Clarithromycin

e) Clindamycin

T F F T T
Mechanism of penicillin
Mechanism of penicillin
Mechanism of penicillin
• Two additional factors are involved in the action of penicillin:

(1) Penicillin binds to a variety of proteins in the bacterial cell membrane and
cell wall, called penicillin-binding proteins(PBPs).

(2) Autolytic enzymes (murein hydrolases) are activated in penicillin-treated

cells and degrade the peptidoglycan. Some bacteria (eg. Stains of
S.aureus) are tolerant to the action of penicillin.
Mechanism of penicillin
Mechanism of penicillin
• Penicillin is bactericidal, but it kills cells only when they are growing.
Penicillins are therefore more active during the log phase.
Forms of penicillin
• Penicillins (and cephalosporins) are called β-lactam drugs because of the
importance of the β-lactam ring
Modification of penicillin structure
Modification of penicillin structure

• Access of the enzyme to the β-lactam ring is blocked by modification of the

side chain with the addition of large aromatic rings containing bulky methyl
or ethyl groups (methicillin, oxacillin, nafcillin).

• Another defense against β-lactamases is inhibitors such as clavulanic acid,

tazobactam, sulbactam, and avibactam.
Activity of Selected Penicillins
Drug Clinically Useful Activity

Gram-positive cocci, gram-positive rods, Neisseria, spirochetes such as Treponema pallidum, and many anaerobes
(except Bacteroides fragilis) but none of the gram-negative rods listed below
Penicillin G

Ampicillin or Certain gram-negative rods, such as Haemophilus influenzae, Escherichia coli, Proteus, Salmonella, and Shigella but not
Pseudomonas aeruginosa or Klebsiella pneumoniae
amoxicillin

Ticarcillin P. aeruginosa, especially when used in synergistic combination with an aminoglycoside

Piperacillin Similar to ticarcillin but with greater activity against P. aeruginosa and Klebsiella pneumoniae

Nafcillin or dicloxacillin
Penicillinase-producing Staphylococcus aureus
 Adverse effect of penicillin

• The major disadvantage of these compounds is hypersensitivity, with a reported

prevalence of 1% to 10% of patients.

• The immunoglobulin (Ig) E–mediated hypersensitivity reactions include

anaphylactic shock, bronchospasm, and urticarial rash.

• IgG and cell-mediated hypersensitivity reactions include nonurticarial skin rashes,

hemolytic anemia, nephritis, and drug fever.
Adverse effect of penicillin
How to prevent penicillin allergy
Inhibition of cell wall synthesis-Cephalosporins
Activity of Selected Cephalosporins
 Adverse effect of cephalosporins

• Cephalosporins are effective against a broad range of organisms, are

generally well tolerated, and produce fewer hypersensitivity reactions than
do the penicillins.

• Despite the structural similarity, a patient allergic to penicillin has only

about a 10% chance of being hypersensitive to cephalosporins also.
 How to prevent drug inactivation

• The inactivation of cephalosporins by β-lactamases (cephalosporinases) is

an important clinical problem.

• β-Lactamase inhibitors such as tazobactam and avibactam are combined

with certain cephalosporins to prevent inactivation of the cephalosporin.
Inhibition of cell wall synthesis-Carbapenems
Carbapenems are β-lactam drugs that are structurally different from
penicillins and cephalosporins.
 Inhibition of cell wall synthesis-Carbapenems
It has excellent bactericidal activity against many gram-positive, gram-
negative, and anaerobic bacteria.

 It is effective against most gram-positive cocci (e.g., streptococci and

staphylococci),

 most gram-negative cocci (e.g., Neisseria),

 many gram-negative rods (e.g., Pseudomonas, Haemophilus, and

members of the family Enterobacteriaceae such as E. coli),

 and various anaerobes (e.g., Bacteroides and Clostridium).

 Inhibition of cell wall synthesis-Carbapenems

• It is especially useful in treating infections caused by gram-negative rods

that produce extended-spectrum β-lactamases(ESBL) that make them
resistant to all penicillins and cephalosporins.

• Carbapenems are often the “drugs of last resort” against bacteria resistant
to multiple antibiotics and are thus reserved for hospital settings.
 Inhibition of cell wall synthesis- Monobactams
• Monobactams (eg.Aztreonam) are also β-lactam drugs .

• It is very useful in patients who are hypersensitive to penicillin

because there is no cross-reactivity.
Inhibition of cell wall synthesis-Vancomycin

• Vancomycin is a glycopeptide that inhibits cell wall peptidoglycan

synthesis by blocking transpeptidation
Inhibition of cell wall synthesis-Vancomycin
• Vancomycin binds directly to the d-alanyl-d-alanine portion of the
pentapeptide, which blocks the transpeptidase from binding,
whereas the β-lactam drugs bind to the transpeptidase itself.
Inhibition of cell wall synthesis-Vancomycin

• Vancomycin is a bactericidal agent effective against certain gram-

positive bacteria.

• Its most important use is in the treatment of infections caused by S.

aureus strains that are resistant to the penicillinase-resistant
penicillins such as nafcillin and methicillin (e.g., methicillin-resistant
S. aureus [MRSA]
Inhibition of protein synthesis
Inhibition of protein synthesis
Mode of Action of Antibiotics That Inhibit Protein
Synthesis
Spectrum of Activity of Antibiotics That Inhibit
Protein Synthesis
Inhibition of nucleic acid synthesis
• Inhibition of Precursor Synthesis - Sulfonamides

• Sulfonamides are structural analogues of p-aminobenzoic acid

(PABA)

• Sulfonamides compete with PABA for the active site of the enzyme
dihydropteroate synthetase
Inhibition of nucleic acid synthesis
Inhibition of nucleic acid synthesis
• Inhibition of DNA Synthesis – Fluoroquinolones

• Fluoroquinolones are bactericidal drugs that block bacterial DNA

synthesis by inhibiting DNA gyrase (topoisomerase)
 Inhibition of nucleic acid synthesis
• Inhibition of mRNA Synthesis- Rifampin

• Rifampin is used primarily for the treatment of tuberculosis in combination

with other drugs.
Mode of Action and Activity of Selected Nucleic
Acid Inhibitors
Alteration of cell membrane function
• Polymyxins

• Daptomycin
Chemoprophylaxis
Principles of antibiotic resistance-mechanisms of
drug resistance
Principles of antibiotic resistance
• Most drug resistance is due to a genetic change in the organism,
either a chromosomal mutation or the acquisition of a plasmid or
transposon.

• high-level resistance

• Low-level resistance
Medically Important Bacteria That Exhibit Significant
Drug Resistance
Genetic basis of resistance

Chromosome-Mediated Resistance
Genetic basis of resistance-plasmid-mediated
resistance
Plasmid-mediated resistance is very important from a clinical point of
view for three reasons:

• (1) It occurs in many different species, especially gram-negative rods.

• (2) Plasmids frequently mediate resistance to multiple drugs.

• (3) Plasmids have a high rate of transfer from one cell to another,
usually by conjugation.
Resistance plasmids (resistance factors, R factors)
Genetic basis of resistance-
• Transposon-Mediated Resistance
 Specific mechanisms of resistance
Penicillins & Cephalosporins:

Cleavage by β-lactamases (penicillinases and cephalosporinases)

Changes in the penicillin-binding proteins (PBPs)

Poor permeability of the drug

Tolerance
 ESBL
• Extended-spectrum β-lactamases (ESBLs) inactivate extended-
spectrum cephalosporins (second- and thirdgeneration cephalosporins)

• Carbapenems, such as imipenem, are the drug of choice to treat

infections caused by ESBL-producing bacteria.
Nongenetic basis of resistance
• Walled off within an abscess

• Resting state

• Protoplasts

• The presence of foreign bodies

• Administration of the wrong drug or the wrong dose

Selection of resistant bacteria by overuse & misuse
of antibiotics
• Use multiple antibiotics unnecessarily.

• Antibiotics are used in animal feed.

Previous year question
• The following statements regarding mechanism of development of resistance are
correct

A) By increasing uptake

B) By conformational alteration in the binding site

C) By synthesizing enzyme

D) By forming new cell wall

E) By developing efflux mechanism

F T T F T
 Principles of pathogenesis
• Pathogen is capable of causing disease.

• Opportunistic pathogens are those that rarely causes disease.

• Virulence is a quantitative measure of pathogenicity and is measured by the number of organisms

required to cause disease.

• The infectious dose of an organism required to cause disease varies greatly among the
pathogenic bacteria. It depends on their virulence factors (eg: pilli, toxin production, capsule,
survival ability against host environment).
Principles of pathogenesis
• Carrier: A person or animal with asymptomatic infection that can be transmitted to
another susceptible person or animal.

• Infection: Multiplication of an infectious agent within the body.

• Invasion: The process whereby bacteria, animal parasites, fungi, and viruses enter
host cells or tissues and spread in the body

• Microbiota: Microbial flora harbored by normal, healthy individuals.

 Principles of pathogenesis
• Nonpathogen: A microorganism that does not cause disease; may be part of the
normal microbiota.

• Superantigens: Protein toxins that activate the immune system by binding to

major histocompatibility complex (MHC) molecules and T-cell receptors (TCR)
and stimulate large numbers of T cells to produce massive quantities of
cytokines
Principles of pathogenesis
Why do people get infectious diseases?

• People get infectious diseases when microorganisms overpower our

host defenses.

• From the organism’s perspective, the two critical determinants in

overpowering the host are the number of organisms to which the host,
or person, is exposed and the virulence of these organisms.
TYPES OF BACTERIAL INFECTIONS

• Bacteria cause disease by two major mechanisms:

 toxin production and

 invasion and inflammation

Identifying bacteria that cause disease

In 1884, robert koch proposed a series of postulates that have been applied broadly
to link many specific bacterial species with particular diseases.

• The microorganism should be found in all cases of the disease in question, and its
distribution in the body should be in accordance with the lesions observed.

• The microorganism should be grown in pure culture in vitro (or outside the body of
the host) for several generations.

• When such a pure culture is inoculated into susceptible animal species, the typical
disease must result.

• The microorganism must again be isolated from the lesions of such experimentally
produced disease
Koch’s postulates
Limitation of koch’s postulate
• Asymptomatic or subclinical infection carriers are now known to be a
common feature of many infectious diseases, especially viral diseases such
as polio, herpes simplex, HIV/AIDS, and hepatitis C.

• The particular bacteria cannot be “grown in pure culture” in the laboratory.

Treponema pallidum (Syphillis)

Mycobacterium leprae ( leprosy)

Virus and rickettsial organism

 Limitation of koch’s postulate
• Not all organisms exposed to an infectious agent will acquire the
infection

• Polymicrobial infection

• Single organism can cause multiple disease

Stages of bacterial pathogenesis

• A generalized sequence of the stages of infection is as follows:

(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, and 5.

(7) Progression or resolution of the disease.

Determinants of bacterial pathogenesis

• Transmission:

Interrupting the chain of transmission is an excellent way to prevent

infectious diseases.

What is vertical transmission and horizontal transmission?

Important Modes of Transmission
Vertical Transmission of Some Important Pathogens
Portals of Entry of Some Common Pathogens
Bacterial Diseases Transmitted by Foods
Zoonotic Diseases Caused by Bacteria
Adherence to Cell Surfaces

• Certain bacteria have specialized structures (e.g., pili)

• or produce substances (e.g., capsules or glycocalyces) that allow

them to adhere to the surface of human cells, thereby enhancing
their ability to cause disease.

• The various molecules that mediate adherence to cell surfaces

are called adhesins.
Pilli
Capsule
Biofilm
• After the bacteria attach, they often form a protective matrix called a
biofilm consisting of various polysaccharides and proteins in
prosthetic surface.

• Biofilms protect bacteria from both antibiotics and host immune

defenses such as antibodies and neutrophils.

• Quorum sensing…
Biofilm
 Invasion, Inflammation, & Intracellular Survival

• Bacteria causes disease by

 Invasion of tissue followed by inflammation.

toxin production

Immunopathogenesis
Invasion
Several enzymes secreted by invasive bacteria play a role in
pathogenesis. Like

• Collagenase and hyaluronidase.

• Coagulase

• Immunoglobulin proteases.
Coagulase, Collagenase and
hyaluronidase.
Limiting host defence and phagocytosis

• Capsule

• M protein and protein A.

• Leukocidins.
Types of bacterial infection

• Bacteria can cause two types of inflammation:

• Pyogenic and granulomatous.

Surface Virulence Factors Important for Bacterial Pathogenesis
Toxin production
• Exotoxins are produced by several gram-positive and gram negative
bacteria

• Endotoxins, which are present only in gram-negative bacteria.

• The essential characteristic of exotoxins is that they are secreted by

the bacteria, whereas endotoxin is a component of the cell wall.
Toxin
Exotoxin

• Exotoxins are poly peptides whose genes are frequently

located on plasmids or

• Lysogenic bacterial viruses (bacteriophages).

Bacteriophages
Bacteriophages
exotoxins encoded by bacteriophage DNA are

• diphtheria toxin,

• cholera toxin, and

• botulinum toxin.
Exotoxin vs endotoxin
Important Bacterial Exotoxins
Previous year question

• Toxins act as superantigen are

A) Erythrogenic toxins

B) Toxic shock syndrome toxin

C) Staphylococcal enterotoxin

D) Botulinum toxin

E) Cholera toxin

T T T F F
Important Mechanisms of Action of Bacterial
Exotoxins
Previous year question
• Following are toxin mediated disease

A) Scarlet fever

B) Plague

C) Dengue

D) Staphylococcal food poisoning

E) Diptheria

T F F T T
Mode of action of Escherichia coli and Vibrio cholerae enterotoxins
 Endotoxins
• The toxicity of endotoxins is low in comparison with that of
exotoxins.

• All endotoxins produce the same generalized effects of fever

and shock

• Endotoxins are weakly antigenic.

Mechanism of endotoxin
Mechanism of endotoxin
 Mechanism of endotoxin

Many activities of lipopolysaccharide (LPS). This bacterial endotoxin activates almost every immune
mechanism, as well as the clotting pathway, which together make LPS one of the most powerful immune
stimuli known. DIC, Disseminated intravascular coagulation
Mechanism of endotoxin
The biologic effects of endotoxin
• Fever due to the release by macrophages of IL-1 (endogenous
pyrogen) and IL-6

• (2) Hypotension, shock, and impaired perfusion of essential

organs due to nitric oxide–induced vasodilation, TNF-induced
increased capillary permeability, bradykinin-induced vasodilation,
and increased capillary permeability
The biologic effects of endotoxin
• Disseminated intravascular coagulation (DIC) due to activation of the
coagulation cascade.

• 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.
The biologic effects of endotoxin
• The end result of the above five processes is called the
systemic inflammatory response syndrome, or SIRS. The most
common clinical signs of SIRS are fever, hypotension,
tachycardia, tachypnea, and leukocytosis.

• Damage to the vascular endothelium plays a major role in both

the hypotension and DIC seen in septic shock.
Previous year question
• Endotoxin is

A) Composed of glycoprotein

B) Firmly bound to the cell surface

C) Heat labile

D) Responsible for causing fever and hypoglycemia

E) Released after death of the bacteria

F T F F T
Beneficial and Harmful Effects of TNF
Different Strains of Bacteria Can Cause Different
Diseases
Host defense

Host defenses are composed of two complementary, frequently

interacting systems:

(1)Innate (nonspecific) defenses, which protect against

microorganisms in general; and

(2) Adaptive (specific) immunity, which protects against a particular

microorganism
Host defense

• Innate defenses can be classified into three major categories:

• (1) physical barriers, such as intact skin and mucous membranes;

• (2) phagocytic cells, such as neutrophils, macrophages, and natural

killer cells; and

• (3) proteins, such as complement.

Host defense
Adaptive immunity is mediated by

• Antibodies and

• T lymphocyte
Host defense
There are two main types of host defenses against bacteria:

• the pyogenic response and the

• granulomatous response.
Host responses to bacterial infection
Inflammation
Essential Host Defense Mechanisms Against Bacteria
 Previous year question
 Cell mediated immunity in the main host defense against -

• Klebsiella spp.

• Pneumocystitis Jiroveci

• Mycobacterium tuberculosis

• Candida albicans

• Streptococcus pyogenes

F T T T F
 Failure of host defenses predisposes to infections
• Certain diseases and anatomic abnormalities also predispose to infections
For example, patients with diabetes often have S. aureus infections

• Patients with sickle cell anemia often have Salmonella osteomyelitis.

• Patients with certain congenital cardiac defects or rheumatic valvular

damage are predisposed to endocarditis caused by viridans streptococci.

• Patients with reduced host defenses often have a muted response to

infection
Conditions That Predispose to Infections
Blood Cultures
• Blood cultures are performed most often when sepsis, endocarditis,
osteomyelitis, meningitis, or pneumonia is suspected.

• The bacteria most frequently isolated from blood cultures are two gram-
positive cocci, Staphylococcus aureus and Streptococcus pneumoniae, and
three gram-negative rods, Escherichia coli, Klebsiella pneumoniae, and
Pseudomonas aeruginosa.

• Certain pathogenic fungi including yeast (Candida species and Cryptococcus

neoformans) and molds can also be isolated from blood cultures
Blood Cultures
Previous year question
• Blood culture may be positive in the following conditions

A) Bacillary dysentery

B) Meningococcal meningitis

C) Primary syphilis

D) Enteric fever

E) Rheumatic fever

F T F T F
Sputum Cultures

• Sputum cultures can be performed to determine infectious etiologies

of pneumonia or to test for active pulmonary tuberculosis.

• The most frequent bacterial cause of community-acquired

pneumonia is S. pneumoniae, whereas S. aureus and gram-negative
rods, such as K. pneumoniae and P. aeruginosa, are common
causes of hospital-acquired pneumonias.
 Cerebrospinal Fluid Cultures
• Cerebrospinal fluid (CSF) cultures can be performed primarily when a

neurologic infection such as meningitis, meningoencephalitis, or transverse

myelitis is suspected. Sample should be sent immediately to laboratory.

• The most important causes of acute bacterial meningitis are three

encapsulated organisms: Neisseria meningitidis, S. pneumoniae, and

Haemophilus influenzae.
Cerebrospinal Fluid Cultures
• If meningitis from acid-fast bacteria such as Mycobacterium
tuberculosis is suspected, acid fast stains of CSF should be performed.

• The fungus Cryptococcus neoformans, a cause of meningitis,

particularly in human immunodeficiency virus–infected patients, can
also be cultured from CSF.
Urine Cultures
• Urine cultures are performed primarily when pyelonephritis or cystitis is
suspected.

• By far the most frequent cause of urinary tract infections is E. coli. Other
common agents are Enterobacter, Proteus, and Enterococcus faecalis.

• To avoid contamination of organisms, a midstream specimen, voided after

washing the external orifice, is used for urine cultures.

• In special situations, suprapubic aspiration or catheterization may be used.

 Urine Cultures
• It is commonly accepted that a bacterial count of at least 100,000/mL must
be found to conclude that significant bacteriuria is present (in asymptomatic
persons). There is evidence that a bacterial count as low as 1000/mL is
significant in symptomatic patients.

• For this determination to be made, quantitative or semiquantitative cultures

are performed.
Culture-Based Methods
 Bacterial vaccine
• Immunity to certain bacterial diseases can be induced either by immunization
with bacterial antigens (active immunity) or by administration of preformed
antibodies (passive immunity).

• Active Immunity
Active immunity can be achieved by vaccines consisting of
(1) bacterial capsular polysaccharides, toxoids, whole bacteria (either killed or live,
attenuated) or
(2) purified proteins isolated from bacteria
 Bacterial vaccine
• Vaccines containing capsular polysaccharide as the immunogen are directed
against
S. pneumoniae,
H. influenzae,
N. meningitidis, and
S. typhi.
• The capsular polysaccharide in the pneumococcal vaccine, the meningococcal
vaccine, and the H. influenzae vaccine is conjugated to a carrier protein to
enhance the antibody response.
Bacterial vaccine
• Two vaccines contain toxoids as the immunogen, the vaccines against
diphtheria and tetanus.

• A toxoid is an inactivated toxin that has lost its ability to cause disease but
has retained its immunogenicity

• Three vaccines contain purified bacterial proteins as the immunogen. The

most commonly used is the acellular pertussis vaccine, which in
combination with diphtheria and tetanus toxoids is recommended for all
children
Current Bacterial Vaccines
Previous year question
• Vaccines are available against following bacteria

• A) Hemophillus influenza type-B

• B) Listeria monocytogens

• C) Streptococcus pneumonia

• D) Staphylococcus aureus

• E) Streptococcus pyogens

• TFTFF
Vaccines Recommended for Children Age 0–6 Years
 Passive Immunity and Passive–Active Immunity

• Passive immunity in the form of antitoxins is available for the

prevention and treatment of tetanus, botulism, and diphtheria.

• This involves providing both immediate (but short-term) protection in

the form of antibodies and long-term protection in the form of active
immunization.

• An excellent example of the use of passive–active immunity is the

prevent of tetanus in an unimmunized person who has sustained a
contaminated wound.