ANAEROBES AND PSEUDOMONAS -

OPPORTUNISTIC INFECTIONS

Faculty: Dr. Alvin Fox

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 MAJOR POINTS

Overview of anaerobic bacteriology

Anaerobic non-spore formers

Anaerobic spore formers

(clostridia)

Pseudomonas (a strict aerobe)

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 KEY WORDS
• Obligate anaerobes • C. perfringens
• Strict anaerobes • Lecithinase
• Polymicrobic (mixed) (phospholipase, α toxin)
infection • C. perfringens
• Spore formers enterotoxin
• Non-spore formers • C. botulinum
• Bacteroides • Botulinum toxin
• B. fragilis • C. difficile
• Clostridium tetani • C. difficile enterotoxin
• Tetanospasmin • Pseudomonas aeruginosa
• Pyocyanin
• Fluorescein
• Toxin A 3
 Obligate anaerobes
• no oxidative phosphorylation
• fermentation
• killed by oxygen
• lack certain enzymes:
superoxide dismutase
O2-+2H+ H2O2
catalase
H2O2 H20 + O2
peroxidase
H2O2 + NADH + H+ 2H20 + NAD

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Polymicrobic anaerobic infection
• Many species in human flora
• Many grow simultaneously - opportunistic
conditions
• opportunistic growth
–injured tissue
*limited blood/O2

• no growth
– healthy tissues
*high O2 content
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 Polymicrobic anaerobic infection

• Simultaneous infection with facultative anaerobe

–diminishes O2 supply further
– aids growth of obligate anaerobes

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 Endogenous versus exogenous
infection
• Two sources
– normal human flora
• endogenous
– environment (e.g. soil)
• exogenous

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 Source of spore-formers and
non-spore formers
• Spore-formers (clostridia)
– exotoxins
– common in the environment (e.g. soil)
– found in normal flora
• Non - spore-formers
– no exotoxins
– mostly normal flora

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 Sites of anaerobes in
normal flora

• intestine
– major site
– 95-99% total bacterial mass
• mouth
• genitourinary tract

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 Bacteroides fragilis
• minor component of gut flora
• most common (strict) anaerobic
infection after abdominal
surgery

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• Enterobacteriaceae (facultative anaerobes)
– commonly cause disease
– low numbers gut flora

• Strict anaerobes
– much less commonly cause disease
– high numbers gut flora

.
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 Strict anaerobe infectious
disease
• Sites throughout body
• Muscle, cutaneous/sub-cutaneous
necrosis
• abscesses

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 Problems in identification of
anaerobic infections
• air in sample (sampling, transportation)
– no growth

• identification takes several days or longer

– limiting usefulness

• often derived from normal flora

– sample contamination can confuse

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 LABORATORY
IDENTIFICATION
• BIOCHEMICAL KITS
- e.g. substrate utilization

• GAS CHROMATOGRAPHY
– volatile fermentation products

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ANAEROBIC NON-SPORE-FORMERS
OF CLINICAL IMPORTANCE
Gram-negative rods:Bacteroides e.g. B. fragilis
Fusobacterium,
Porphyromonas, Prevotella
Gram-positive rods:
Actinomyces, Bifidobacterium, Eubacterium
Lactobacillus, Mobiluncus, Propionibacterium
Gram-positive cocci: Peptostreptococcus and Peptococcus

Gram-negative cocci: Veillonella

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 Bacteroides fragilis
• Major disease causing strict anaerobic
non-spore-former
• Prominent capsule
– anti-phagocytic
– abscess formation

• Endotoxin
– low toxicity
– structure different than other
lipolysaccharide 16
ANAEROBIC SPORE-FORMERS
(CLOSTRIDIA)

• Gram-positive rods
– human intestine
– soil

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 Clostridium tetani

spore

vegetative
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 Clostridium tetani
• Non-invasive

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 Tetanospasmin
• disseminates systemically
• binds to ganglioside receptors
– inhibitory neurones in CNS
• muscles keep on working
• spastic (rigid) paralysis
• glycine
– neurotransmitter
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A severe case of tetanus.
muscles, back and legs are rigid
muscle spasms can break bones 21
can be fatal (e.g respiratory failure)
 Vaccination

• infant
• DPT (diptheria, pertussis, tetanus)
• tetanus extremely uncommon in US
• tetanus toxoid
– antigenic
– no exotoxic activity

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 C. perfringens
• soil, fecal contamination
• war
• gas gangrene
– swelling of tissues
– gas release
* fermentation products
• wound contamination

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 Pathogenesis
• tissue degrading enzymes
– lecithinase [α toxin]
– proteolytic enzymes
– saccharolytic enzymes

• destruction of blood vessels

• tissue necrosis
• anaerobic environment created
• organism spreads

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Without treatment death
occurs within 2 days

 effective antibiotic therapy

 debridement
 anti-toxin
 amputation & death is rare

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 Laboratory identification

• lecithinase production

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 Food poisoning
• enterotoxin producing strains

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C. botulinum

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 Botulism
• food poisoning
– rare
– fatal

• germination of spore
• inadequately sterilized canned food
– home
• not an infection

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 Botulinum toxin
• binds peripheral nerve receptors
– acetylcholine neurotransmitter
• inhibits nerve impulses
• flaccid paralysis
• death
– respiratory
– cardiac failure

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 Infection with C. botulinum
• Neonatal botulism
– uncommon
– the predominant form of botulism
– colonization occurs
• no normal flora to compete
• unlike adult

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 Wounds

– extremely rare
– an infection

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 Botulinum toxin
• Bioterrorism
– not an infection
– resembles a chemical attack

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 Treatment
• anti-toxin
• antibiotic therapy (if infection)

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 C. difficile
• After antibiotic use
• intestinal normal flora
– greatly decreased
• colonization occurs
• enterotoxin secreted
• pseudomembanous colitis

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 Therapy
• discontinuation of initial
antibiotic (e.g. ampicillin)

• specific antibiotic therapy (e.g.

vancomycin)

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PSEUDOMONAS AERUGINOSA

Gram negative rod

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 Pseudomonas
• Aerobic
• Gram-negative rod
• majority of human infections
– P. aeruginosa

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 Common in the environment
• water
• air
• soil

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P. aeruginosa and compromised host
• Burns and wounds
– destruction of blood vessels
– phagocyte access limited
•cancer
– cytotoxic drugs
*destroy the immune system
•cystic fibrosis
– altered respiratory epithelium
– pneumonia

. 40
 Identification
• Pigments
– pyocyanin (blue-green)
– fluorescein (green-yellow, fluorescent)

• biochemical reactions

• cultures have fruity smell

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 Pathogenesis
• Slime layer is anti-phagocytic

• Toxin A - ADP ribosylates EF2

– similar to diphtheria toxin

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