ANAEROBES

Objectives
At the end of the lecture the students should have
understood and be able to:

 Describe the general characteristics of anaerobes

 Describe their simple classification into sporing
and non-sporing anaerobes
 List the organisms, their infections and diseases
 Describe the laboratory investigations
 Discuss the treatment options and prevention
General Characteristics
 Intolerant of molecular oxygen
 Lack catalase
 Lack of superoxide dismutase
 Require low oxidation-reduction
potential or redox potential (Eh) -
mV
 Uniform resistance to
aminoglycosides
 Nearly all are sensitive to
 Classification
 Sporing:
– Gram-positive e.g clostridia
 Non-sporing:
– Gram-positive e.g. Bifidobacterium,
Propionibacterium
– Gram-negative e.g. Bacteroides,
Fusobacterium, Porphyromonas,
Prevotella,
 CLOSTRIDIA
 Gram-positive spore-forming
anaerobic bacilli
 All produce spores
 Biochemically active, -
saccharolytic and proteolytic
properties
 Neurotoxins - C. tetani and C.
botulinum
 Aggressins - gangrene - C.
perfringens soft tissue.
Clostridium perfringens
 Large Gram-positive bacilli
 Capsulated
 Non-motile
 Identified by Nagler reaction
 Produces double zone haemolysis on
blood agar
 Heat-resistant spores - food
poisoning
 Heat-labile spore - gas gangrene)
 α-toxin - a phospholipase C.
 Gas gangrene
 Oedema, myositis, necrosis of
tissue, gas production and severe
toxemia

 Source - animal and human faeces

 Impairment of blood supply,

reduction in O2 supply and
colonization
 Treatment
 Surgery - debridement
 Remove all sutures, and excision
 Penicillin, metronidazole and
aminoglycoside in combination.
 Clindamycin plus an aminoglycoside or
imipenem are good alternatives
 Hyperbaric oxygen
 Polyvalent antiserum (C. perfringens, C.
septicum and C. novyi antitoxins).
 Food poisoning
 Heat-resistant spore
 “Heat-shock”
 Sporulation in the intestine –
enterotoxin
 Abdominal cramps 8-12 hours after
ingestion, followed by diarrhoea
 Symptoms usually subside within a day
or two
 Source - a precooked meat food.
 Diagnosis - isolation of similar strains in
the faeces of victims and the food itself
 Clostridium tetani
 Straight, slender Gram-positive bacilli
with rounded ends (drumstick
appearance)
 Strict anaerobe
 Motile
 Produces a thin spreading film on
enriched BA
 Spore resistant to boiling, dry heat, 5%
phenol
 Toxins – Tetanolysins &
 Tetanus (Lockjaw)
 Wound – RTA - C. tetani spore
 Toxin - to central nervous system
via motor nerve:
– Specifically by the alpha-motor fibres.
– Some via the blood
 Inhibition of motor impulses
– produced by the upper motor neuron
over the lower,
– producing an increase in tonus and
tonic spasms.
 Tetanus (Lockjaw)
 Acute neuromuscular disease
– Tetanospasmin
– Painful muscular contractions –
masseter and neck muscles
– Spasms
– Rigid paralysis
– Respiratory failure
– Death
 Incubation period: 3 – 21 days
Other Forms of Tetanus
 Tetanus neonatorum
– Follows infection of the umbilical
wound of newborn infants

  Postoperative tetanus
– Imperfectly sterilized catgut,
dressings or glove powder
– Dust-borne infection of the wound at
operation.
Prevention and control
 Prompt and adequate wound toilet
 Proper surgical debridement
 Penicillin
 Antitoxin
– human tetanus immunoglobulins
(HTIG; homologous antitoxin;
dose 250-500 units)
– equine antitoxin (ATS;
heterologous)
– Specific immunization with tetanus
Prevention and control
 Active
immunization with DTP
vaccine

 Booster
– 10 years interval
– After wound or accident

 HTIG to non-immunized persons

following puncture wound
Clostridium botulinum

 Motile
 Sporeoval and subterminal
 Saprophyte
– soil, vegetables, fruits, leaves,
silage, manure, muds
 Produces potent neurotoxin in
food.
 Botulism
• Food poisoning with neurotoxic
effects
• Associated with a variety of foods:
 preserved ham, large sausages,
home-preserved meats and
vegetables, canned products such as
liver paste, fish, hazelnut puree
• Preformed toxin in the food is
absorbed from the GIT
 affects the cholinergic system,
blocking the release of acetylcholine
in the peripheral nervous system
 Clostridium difficile
 Motile
 Normal flora GIT of neonates and
infants
 Hardly found normal healthy adults.
 Toxins: Toxin A (an enterotoxin)
and toxin B (a cytotoxin).
 C. difficile-associated diarrhoea
(CDAD)
– Antibiotic-induced diarrhoea,
– Antibiotic-associated colitis
– Pseudomembranous colitis.
 C. difficile-associated
diarrhoea (CDAD)
 Antibiotic-induced diarrhoea
– Mild, self-limiting
 Antibiotic-associated colitis
– Moderately severe
 Pseudomembranous colitis.
– Very severe
– Bloody
– Associated with loss of protein
– High mortality
 C. difficile-associated
diarrhoea (CDAD)
 Affects the elderly and geriatric patients
 Patients in the ICUs, geriatric and surgical
wards
 Follows use of antibiotics e.g. clindamycin,
cephalosporins, etc
 Treatment
– Metronidazole
or
– Vancomycin
 Clostridium difficile
Predisposing factors
 Previous antibiotic use:
– clindamycin, cephalosporins, ampicillin and
nearly all antibiotics may cause both
diseases.
 Surgery
 Nasogastric tube
 Hospitalized patients particularly long-
stay and geriatric patients
 NON-SPORING
ANAEROBES (NSA)
 Most frequently encountered
anaerobes in clinical medicine.
 No spores
 Part of the normal flora.
 Found in the oropharynx,
gastrointestinal tract, female
genital tract
 Colon - 1012 anaerobes per gram
of faeces
 NON-SPORING
ANAEROBES
In the colon
– Anaerobes:aerobes = 1000:1.
In the mouth
– Anaerobes:aerobes = 100:1.
In the vagina
– Anaerobes:aerobes = 100-1000:1.
 Theyusually occur in mixed flora
in many infections
Non-sporing anaerobes (NSA)
Predisposing factors:
 Immunological impairment
 Decrease in host defense systems
 Tissue damage
 Devitalization,
 Surgery
 low Eh.
Clinical signs and clues
 Foul smelling pus, discharge or
lesion
 Large amount of pus (abscess
formation)
 Proximity of lesion to mucosal
surface or portal of entry
 Infection associated with necrotic
tissues
 Deep-seated abscesses
 Gas formation in tissues
 Examples of NSA
 Bacteroides species, - B. fragilis
 Fusobacterium species, - F. nucleatum
 Prevotella species (majority are black
pigmented), - Prev. melaninogenica
 Porphyromonas spp. (all are black
pigmented) - Porph. gingivalis
 Peptostreptococcus spp., - P. micros
 Actinomyces spp., - A. israelii
 Colonial Morphology
 Bacteroides species, -
B. fragilis
 Fusobacterium
species, -
F. nucleatum
 Black pigmented anaerobes
 Porhyromonas
gingivalis
 All Porhyromonas spp
 Prevotella intermedia
 Prevotella
melaninogenica
 Non-Sporing Anaerobic
Infections
 Brain abscess
 Oro-dental abscesses
 Post-op infection
 Aspiration pneumonia
 Lung abscesses
 Intra-abdominal abscesses
 Liver abscess
 Soft tissue infection –
diabetic ulcers
 B. fragilis, Ano2 cocci
 Porph.gingivalis, Fusobact
 B. fragilis
 Oral Anaerobe, B. fragilis
 Oral Anaerob, Ano2 cocci
 B. fragilis
 B. fragilis, Prev. intermed
 B. fragilis, Clostridia
 Treatment
 Excision of abscess and necrotic tissue,
drainage of pus.
 They are all resistant to
aminoglycosides
 Metronidazole is the drug of choice
 Other anti-anaerobic drugs:
– Clindamycin
– Meropenem
– Imipenem
– Piperacillin-tazobactam
Case 1
A 20 year old man sustained an open fracture of
the tibia as a result of road traffic accident. Two
weeks later he developed clonic and tonic spasm
of the jaw and back muscles. Gram-positive
slender bacilli with terminal spores were isolated
from his infected leg wound.

1. What is the likely diagnosis?

2. What is the causative organism?
3. What initial treatment should he have received?
4. How would you treat this patient?
5. What vaccination regimen is available?
 Case 2
Four days after admission to the intensive care unit
(ICU), a 58-year old man developed profuse watery
diarrhoea following therapy with clindamycin. He
was mildly to moderately dehydrated and was
being fed through a nasogastric tube.

7. What disease syndrome does he have?

8. Which organism is responsible for this disease?
9. Name its main virulence factor
10. Name 2 predisposing factors for the disease
11. How should this patient be treated?