BASIC BACTERIOLOGY

BACTERIA CELL ENVELOPE

FORMS DEFINITION Gram (+) Gram (-)
Coccus THICK peptidoglycan THIN peptidoglycan
(+) Lipoteichoic acid (-) Lipoteichoic acid
in pairs with both - Polymer of glycerol units
Diplococci
apposing sides - Major cell surface antigens
2 layers 3 layers
Cell Membrane Inner Membrane
Tetrads groups of four Cell Wall Periplasmic Layer
Outer Membrane
Antigenic lipopolysaccharide (O-polysaccharide)
Streptococci in chains Toxic lipopolysaccharide (Lipid A)

Staphylococci in grape-like clusters

Bacillus

Coccobacillus very short

CELL WALL CELL MEMBRANE

Filamentous long filaments Peptidoglycan layer Lipid bilayer
Peptido portion: short strings of amino acids Does not differ from human cell membrane

Contains
Fusiform tapered ends Glycan Portion: alternating monosaccharide subunits
-N-acetylglucosamine (NAG) Porins - channels
-N-acetylmuramic acid (NAM) Permeases - carriers for specific substances
club-shaped, comma-
Vibrioid
shaped

Spiral

Spirila helical

PREVENT CELL FROM EXPLODING IN HYPOTONIC

Spirochete corckscrew
SOLUTIONS

TARGET OF ANTIBACTERIAL AGENTS

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 MEDICAL SIGNIFICANCE OF CELL WALL APPENDAGES
Drug Action Flagella Pili or Fimbriae
Inhibit enzymes (Penicillin-binding proteins) that catalyze long, semi-rigid, helical, hollow, tubular
B-Lactam antibiotics Shorter and thinner
transpeptidation and carboxy peptidation reactions structures
Bactiracin Inhibits recycling of bactoprenol phosphate for MOTILITY attachment organs for cell-cell contact
- lipid synthesized by three different species of lactobacilli responds to chemotactic stimulus -Neisseria gonorrhea
Inhibits synthesis of D-ala-D-ala dipeptide that provides two terminal attached to cell wall or membrane by a basal
Cycloserine residues of pentapeptide -E. coli
body that rotates the flagellum
-It is involved in the transpeptidation that crosslinks the peptide side Highly antigenic (K-antigen)
chains of peptidoglycan strands
Blocks incorporation of the NAG-NAM-PEP repeat unit into the growing
Vancomycin peptidoglycan chain

GLYCOCALYX
Capsule Slime Layer
Firmly associated polysaccahride Loosely associated glycoprotein
Thick Thin
Tightly bound to cell wall Loosely bound to cell wall
Types of
Well-organized layer Unorganized layer
Flagella Definition Structure Example
Difficult to be washed off Easily washed off
Aids in adherence and protects cell from Monotrichous single flagellum at ONE POLE V. cholera
Helps evade phagocytosis
dehydration and nutrient loss
Bartonela
Lopotrichous tuft of flagella at ONE POLE bacilliformis

tuft of flagella at BOTH ENDS Spirillum

Amphitrichous serpens
of the cell

distributed ALL
Petritrichous E. coli
THROUGHOUT the cell

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 CYTOPLASMIC STRUCTURES
Cytoplasm Nucleoid Plasmids Ribosomes Inclusions Endospores Mesosomes
mixture of substances for POLYMERATION of Invaginations of
Fibrous mass of Small fraction of DNA Accumulations of food Dormant form within
enclosed by the cell amino acids into bacterial cell
genetic material molecules reserves vegetative cell
envelope proteins membrane
(DNA with singular involved in
consists of amorphous Exists and replicates Can be released as free
chromosome and 70s (50s and 30s) -polysaccharides CHROMOSOME
aqueous fluid AUTONOMOUSLY spores
RNA) SEPARATION
related ot nuclear
Where suspended or No discernible Caryy genes for a Rich in CALCIUM
-lipids envelope of
dissolved are: membrane enclosure variety of functions DIPICOLINATE
eukaryotes
have function similar
to MITOCHONDRIA
NOT ESSENTIAL for
enzymes -polyphosphates Resistant to (membranous support
bacterial viability
for respiratory
enzymes)
Membrane-bound or -Heat (survive
ions Enhance survival by
scattered boiling)
-conveying antibiotic
metabolites -UV light
resistance
-enhancing mating Bacteriocidal
storage granules
ability chemical agents
-making possible Destroyed by
DNA, RNA, proteins
toxin production AUTOCLAVING
plasmids

SPORULATION MEDICAL SIGNIFICANCE OF SPORULATION

 Most notorious pathogens are spore-formers

 Bacillus anthracis (Anthrax)
 Costridium botulinum (Botulism)
 Clostridium perfringens (Gas gangrene)
 Spores can remain viable for many years

SPORE GERMINATION
 Return of spore to vegetative state
 Occurs in nutritionally rich environment
 Process:
1. Destruction of cortex by lytic enzymes
2. Uptake of water
3. Release of dipicolinate from the cell
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 STAGES OF BACTERIAL GROWTH CYCLE VARIOUS WAYS TO MEASURE BACTERIAL GROWTH
Microscopic counting
 Ruled slide

Colony counting
 Agar surface growth

Quantitating turbidity
 Culture in liquid medium

1. Lag Phase
 Bacterial cells acquire nutrients and replicate genetic material
 Preparation for cell division

2. Log Phase or Exponential Phase

 Bacterial reproduction through binary fission
 Exponential increase
 Doubling Time (20mins – several days)
 E. coli – 10M cells in 8 days

3. Stationary Phase
 Growth slow down
 Nutrients are depleted, toxic products accumulate, and
environmental pH is changed
 Multiplication = Cell Death
 Formation of endospores for spore-forming organisms

4. Phase of Decline
 No more cell multiplication
 Release of spores for spore-forming organisms

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 BACTERIAL GROWTH REQUIREMENTS
Nutrient Temperature Oxygen
Facultative Obligate
Heterotrophs Autotrophs Psychrophilic Mesophilic Thermophilic Aerobes
Anaerobes Anaerobes
Obligated
Microaerophilics
aerobes
lack the enzyme
PEROXIDASE and
SUPEROXIDE
Utilize CO2 for utilize oxygen as grow in
All clinically DISMUTASE
the synthesis of Grow best at -5 Grow between Grow between final electron lower oxygen PRESENCE or
important required for
organic to 30deg C 10 to 45deg C 50 to 60deg C acceptor in ATP tension ABSENCE of
bacteria elimination of
metabolites production oxygen
oxygen
metabolism by-
products
Obtain sugar
Includes MOST
from the
BACTERIA
environment M. tuberculosis Neisseriae E. coli
pathogenic to
they are in
human
Require organic
carbon for
growth

Nitrogen, sulfur, phosphorus,

potassium, calcium magnesium, and
iron. Growth factors

GROWTH REQUIREMENTS
Energy pH
Aerobic respiration Anaerobic respiration Fermentation Acidophiles Neutrophiles Alkalinophiles
Molecular oxygen serves as final Inorganic compounds are final
Alternative anaerobic process
electron acceptor electron acceptor <.5 6 to 8 >8.5
Organic metabolic intermediate
Alternative to aerobic respiraion for dervied from a "fermentable"
Oxygen is reduced to water
facultatives substrate us the final electron
acceptor
Used by ALL AEROBIC BACTERIA

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 CULTURE MEDIA
Enrichment Selective Differential
only allow growth of specific bacterial species from
promotes NON-SELECTIVE growth of any bacteria Allows differentiation of organisms by the
specimens that normally contain large bacterial
present in a clinical sample variation of growth patterns
population (stool, genital tract discharges, sputum)

MacConkey - differentiates lactose fermenters

useful in growing FASTIDIOUS ORGANISMS
Contain BLOOD, YEAST, EXTRACT, BRAIN OR HEART
INFUSIONS
Sheep BA - most organisms from human sources
Chocolate agar - lysed RBCs release hemin and NAD
required by fastidioud organisms
Selective growth is due to inhibitory substances
from non-lactose fermenters
MacConkey - most Gram(-) rods, inhibits Gram(+)s
Sheep BA - hemolytic reactions
and fastidious Gram(-)s
Thayer-Martin - for pathogenic Neisseriae
SS - for Salmonella and Shigella

BACTERIAL GENOME
Chromosome Plasmids
Single circular double-stranded DNA Small circular DNA
Contains ESSENTIAL and NON ESSENTIAL genes size is less than 1/10 of chromosome
2000-4000 1500-120000
Replicate INDEPENDENT of the chromosome
May exist as one or many copies within
Hemophilus influenzae (1995) bacterium
1,743 genes (1,830,137 base pairs) may carry GENE FOR TOXINS
May contain mobile DNA sequences called
141 genes - protein synthesis TRANSPOSONS which are repository for many
antibiotic resistance genes
105 - energy metabolism
123 - regulate passage of materials across the cell
membrane

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BACTERIOPHAGE
 Virus that replicates within bacterial cell TYPES OF BACTERIOPHAGE
 Consists of piece of nucleic acid encapsulated in protective protein coat
 May be DNA or RNA type, double or single-stranded
Virulent Temperate
Bacterial infection result in FUSION OR
 Induction
Infection of bacterial cell results in CELL INTEGRATION OF PHAGE DNA INTO HOST
 Emergence of virus from latent to prophage state
DEATH CHROMOSOME (prophage state)
 Lysogenic conversion Expression of prophage gene is repressed by
There is a release of newly replicated phage
 Acquisition of new properties by bacteria due to presence of prophage repressor protein
particles
 Examples 1 phage -> hundreds of progeny phage in
 Corynebacterium diphtheria No new phage particles are formed
20mins
 Spreptococcus pyogene Generally phage infection of bacterial
No host cell lysis occurs
species has SPECIFICITY
Phage DNA is replicated as part of host
Replicative cycle of bacteriophages genome
LYSOGENIC BACTERIA
 CARRIER OF PROPHAGE
 Phenomenon is called LYSOGENY
 Destabilization of prophage-bacteria
 Occurs in exposure to UV light
 Damages host DNA
 Lifting of gene repression
 Leads to production of progeny phage particles and host cell lysis

MECHANISMS OF ACQUIRED ANTIBIOTIC RESISTANCE

Decreased uptake of antibiotic

 by LIMITING THE NUMBER OF PORINS in the outer membrane

Alteration of antibiotic target site

 Exemplified by ALTERATION OF PENICILLIN-BINDING PROTEINS

Acquisition of ability to destroy or modify antibiotic

 Genetic information for B-lactamases, acetyltransferases, and esterases

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Conjugation
cell-cell contact
Requires DONOR and RECEPIENT CELLS
Sex pilus (cytoplasmic bridge) allows DNA transfer
Conjugative plasmids carry genes
-that promote their own transfer to other cells
-that direct synthesis of sex pili
-for antibiotic resistance
GENE TRANSFER
Transduction Transformation
via phage vector by means of naked DNA
-recepient vector is transduced when phage depends on transitory physiologic state that
vector infects and injects content of bacterial gene allows DNA to cross the cell membrane
NO CELL-CELL CONTACT Its discovery provided the first evidence that
DNA IS THE GENETIC MATERIAL INVOLVED (1944)

Capsules
Usually polymers of
polysaccharides
Acquired via TRANSDUCTION
PREVENT PHAGOCYTOSIS
Prevent binding of Ab and C3
on bacterial membrane to
phagocyte receptors
VIRULENCE FACTORS
Adhesins Invasive factors Exoenzymes Toxins
Exotoxins Endotoxins
Collagen and fibrin lysis -
collagenases, hyaluronidases, Capability may be due to
Pili (finbriae) Pili and enzymes fibrinolysins LPS of gram negative bacteria
Invasion plasmid protein of Cell material lysis - proteases,
1. encoded in the genome
LPS Shigella lecithinases Not actively secreted
M protein of S. pyogenes -
-facilitate attachment and 2. from plasmids and Cause SEPSIS and SEPTIC
require receptors in target Antibiotic modification/
subsequent endocytosis bacteriophages SHOCK
cells inactivation - B lactamases
Structure:
1. binding site
2. active domaine
Functional classification
1. Enterotoxins - causes
increased epithelial cell
secretion
2. Neurotoxins - botulinum
toxin and tetanospasmin
3. Cytotoxins - inhibit
cellular metabolism

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