Anaerobes of Clinical Importance

Objectives:
1/ Describe the oxygen relationship with bacteria.
2/ Differentiate obligate, moderate, and aerotolerant anaerobes.
3/ Describe how anaerobes of normal microbiota initiate & establish
infection.
4/ Name the endogenous anaerobes commonly involved in human
infections and the types of infections with which they are associated.
5/ Give the manifestations of infections caused by Clostridium spp. and
classify the genus based on their toxin activities.
6/ Identify the most probable anaerobic causative agents of the
following: wound botulism, tetanus, gas gangrene, actinomycosis, acne,
pseudomembranous colitis.

Võ Thị Chi Mai

 Effect of Oxygen on Bacterial Growth

• Obligate aerobes
• Facultative anaerobes
• Microaerophiles
• Obligate anaerobes: yielding energy
through metabolite fermentation.

Types of anaerobes
▪ Strict anaerobic bacteria
▪ Moderate obligate anaerobes
▪ Aerotolerant anaerobes.
Metabolism in the presence of O2
Intermediate metabolites comprised of toxic forms of oxygen:
• O2- (superoxide)and H2O2 are (hydrogen peroxide) strong
oxidizers.
• OH* (hydroxyl radical) is extremely reactive.

Enzymes neutralizing toxic forms

Catalase, Peroxidase are porphyrin-

containing proteins, & Superoxide
dismutase is metal-containing protein.
These enzymes destroy toxic
intermediate metabolites.
Obligate anaerobes:
- Sensitive to toxic products of oxygen metabolism.
- Require reducing environment low Eh.

 Deficiency or lack of
• superoxide dismutase
• peroxidase
• catalase

 Main enzymes active in reduced condition.

 Anaerobic infection
• Abscess of any organ
• Actinomycosis
• Aspiration pneumonia, necrotizing pneumonia, empyema
• Complication of appendicitis or cholecystitis
• Dental & periodontal infection
• Endocarditis, bacteremia, septicemia
• Meningitis, usually following brain abscess
• Otitis media, sinusitis
• Necrotizing fasciitis, cellulitis, myonecrosis
• Osteomyelitis
• Peritonitis, intra abdominal infections.
 Endogenous anaerobic infections
1/ Gram-neg. bacilli: penicillin-resistant, metronidazole-susceptible
Bacteroides: growth in bile, pigmented colonies
Bacteroides, Prevotella, Porphyromonas
Fusobacterium: very sensitive to oxygen.

Bacteroides fragilis Fusobacterium nucleatum

Pigmented Bacteroides
(Koneman)
 Endogenous anaerobic infections
2/ Gram-pos. cocci: usually penicillin-susceptible
Peptostreptococcus

3/ Non-sporeforming G+ bacilli: (Koneman)

Slow growth. Penicillin-S.Metronidazole-R.

Cutibacterium, Actinomyces; Bifidobacterium, Lactobacillus
 Endogenous anaerobic infections

4/ G- cocci:
Veillonella: putrescin, cadaverin as required growth factors

Veillonella dispar
Koning - thesis
 Pathogenesis
• Trauma: Decreased oxygenation of tissue.
• Bacterial synergism.
• Increased resistance of anaerobes, access to deeper tissues.
• Consequences of therapy: X-ray, radiation, immunosuppressive...

Indications of involvement of anaerobes in infections

• Infection in close proximity to a mucosal surface.
• Presence of foul odor, large quantity of gas.
• Presence of black color / brick red fluorescence.
• Presence of “sulfur granules”.
• No growth in aerobic culture.
Endogenous anaerobic infections

Noma

Actinomycosis
Ludwig’s angina

Diabetic foot
Cellulitis

(Upjohn)
 Exogenous anaerobic infections
(Prescott)

Genus Clostridium:
Spore-forming G+ bacilli, producing exotoxins.
1/ neurotoxins
2/ toxins causing gas gangrene
3/ antibiotic-associated colitis

Clostridium botulinum Clostridium tetani

(Hart)
 Exogenous anaerobic infections
1a/ Clostridium botulinum: botulism
• botulinum toxin attaches to the neuromuscular junction of nerves.
• prevents the release of acetylcholine  flaccid paralysis & death.

1b/ Clostridium tetani: tetanus

• tetanospasmin binds to gangliosides.
• blocks inhibitory impulses to the motor neurons  Spastic type of
paralysis.
 Exogenous anaerobic infections
2/ Clostridium perfringens:
produces various exotoxins:
• -toxin → cellulitis, myonecrosis
• -toxin → necrotizing enteritis
• enterotoxin → food poisoning (diarrhea).
And other clostridia.
 Bauman

Newborn tetanus Risus sardonicus

Infantile botulism

Cellulitis Myonecrosis (gas gangrene)

Necrotizing enteritis
Mims
 Exogenous anaerobic infections

3/ Clostridioides (Clostridium) difficile:

• Toxin A (TcdA = enterotoxin) + toxin B (TcdB = cytotoxin)
• Antibiotic-associated colitis
• Pseudomembranous colitis

Yellow plaques (sigmoidoscope)

 Anaerobic diagnosis in Lab
Bacteriological diagnosis: Pouch
• Morphology
Axonomat
• Culture – Toxin assay
• Biochemical characteristics for identification
• Antibiotic susceptibility testing
• Gas-liquid chromatography
• Molecular methods: PCR
• MALDI-TOF Glove box

Treatment
• Surgical procedure – Antimicrobial – Hyperbaric oxygen.
• Antitoxins (tetanus & botulism).
• Fecal microbiota transplant.
 Summary

• Endogenous anaerobic infections: opportunistic inf.

• Exogenous anaerobic infections: due to exotoxins.