Kochs Postulates

The organism must be isolated in all cases

of a designated disease and not from
healthy animals
The organism must be isolated from the
infected person (or animal) and grown in
pure culture
The organisms from the pure culture must
reproduce the disease when inoculated in a
susceptible animal
The organism then must be isolated from
the
Canexperimentally
you think of someinfected
shortcomings?
animal

Lipopolysaccharide=
lipid+polysaccharide

O-antigen required
for virulence,
protects against
phagocytosis, killing
by complement

Lipid A = endotoxin
Saturated fatty acids
pack tightly to decrease
permeability to
hydrophobic agents

Porin forms trimer in bacterial outer membrane.

Pore is ~10A, and allows passage of H2O,
amino acids, monosaccharides, disaccharides
Monomer(barrel)
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trimer
May
exclude
antibiotics,
toxic
chemicals,
lysozyme
Contributes to function of outer membrane as size exclusion barrier

Periplasmic Space
Only in gram negatives
A compartment
Filled with gel of peptidoglycan and water

Degredative enzymes found here

Amylases, nucleases
Break down large nutrients

Detoxifying enzymes found here

Beta-lactamases

Cytoplasmic membrane

Site of metabolic pathways

Sensing the environment

Chemotaxis, signal transduction

Nutrient transport;

Electron transport (respiration)

Proton gradient used for ATP synthesis, flagellar
motion
Cell wall biosynthesis (peptidoglycan, LPS)

Diffusion: passive
Facilitated: specific pore
Symporter or antiporter: exchange
Active transport: uses ATP

Efflux of macromolecules

Peritrichous bacteria: flagellae coalesce into a tail

during one direction of rotation, opposite rotation leads to tumbling

Bacterial Metabolism
Obligate anaerobes: killed by oxygen
Clostridium perfringens: gas gangrene

Obligate aerobes: require oxygen

Mycobacterium tuberculosis: lung TB

Facultative: can grow with or without

oxygen
Escherichia coli

Gram Stain and Culture

Results

Anaerobic cultures
sent
intraoperatively
grew Prevotella

Strict anaerobe

Anaerobes: Clostridium,
Bacteroides, Prevotella,
Fusobacterium
http://pathmicro.med.sc.edu/ghaffar/brucell03-2.jpg
http://www.hull.ac.uk/mouthcare/cal-mouth/mouthcare/images/fig9.jpg

Plasmids and Transposons

B. Transposons
1. Small genetic elements capable of mediating their own
movement from one chromosomal (or plasmid) location to
another
2. Compound transposons carry accessory genes, such
as antibiotic resistance genes

C. Clinical Relevance
1. Mechanism of spread of antibiotic resistance among grampositive and gram-negative bacteria

Conjugation
A. Plasmid-mediated process of DNA transfer
from one cell to another that requires:
1. Cell-to-cell contact and formation of a conjugation
bridge via a sex pilus
2. A conjugal plasmid capable of transfer replication

B. Plasmid transfer by conjugation

1. Conjugative plasmid in donor (male) cell encodes
sex pili to initiate cell-cell contact
2. Transfer replication creates a single-stranded copy
of the plasmid that is transferred to the recipient
(female). Transfer replication provides the
driving force for transfer and is absolutely
required for conjugation.
3. Single-stranded plasmid is converted to a circular
duplex molecule in the recipient (now male)

Bacterial Conjugation
C. Transfer of chromosomal genes by conjugation
1. Happens when the conjugal plasmid is integrated in the host
cell chromosome
2. Transfer replication is initiated within the plasmid sequences
but additional flanking chromosomal DNA is also
conjugated to the recipient
3. The transferred DNA cannot be maintained as an
autonomous plasmid so homologous recombination is
required if the incoming DNA is to be maintained

D. Clinical Relevance
1. Spread of antibiotic resistance from plasmid and
chromosomal loci in both gram-positive and gram-negative
bacteria

Transformation
A. Historical Evidence
1. Griffith: avirulent pneumococcus can be transformed into a
virulent bacterium using a heat-killed extract of the virulent
donor strain.
2. Avery, MacLeod and McCarty: transforming principle is
DNA, because transformation is DNAse-sensitive.
Transformation does not require cell-to-cell contact.

B. Mechanism
1. Development of competence
2. DNA binds to the cell surface
3. DNA enters the recipient cell
4. DNA integrates into host chromosome by homologous
recombination. Plasmid molecules can replicate
extrachromosomally

Bacterial Transformation
C. Clinical Relevance
1. Examples of capsule exchange among pneumococci
2. Most significant is acquisition of genes involved in resistance
to antibiotics
a. Genes that encode antibiotic binding proteins (e.g.,
penicillin binding proteins, gyrase/topoisomerase IV
binding proteins)
b. Genes that encode enzymes that inactivate antibiotics
(e.g., -lactamase)
c. Genes that encode exporters of antibiotics (e.g.,
tetracycline efflux)

Transduction lytics vs lysogenic

D. Transduction
1. Result of occasional packaging of host DNA during lytic infection
and subsequent transfer to a recipient via phage injection
2. Steps in the transduction process
a. Infection of donor strain
b. Donor DNA packaging
c. Infection of recipient
d. Recombination of donor DNA with the recipient chromosome

E. Clinical Relevance
1. Toxin genes can be carried on bacteriophage, e. g.
a. Diphtheria toxin in Corynebacterium diphtheriae
b. Streptococcal erythrogenic toxin
c. Botulinum toxin (Clostridium botulinum)
d. Cholera toxin (Vibrio cholerae)

Mechanisms Conferring Resistance