This document provides an overview of anaerobic microorganisms. It discusses their typical habitats including the skin, oral cavity, gastrointestinal tract, and urogenital tract. It outlines their classification and important genera such as Bacteroides, Prevotella, and Clostridium. Virulence factors and mechanisms of pathogenesis are described. The laboratory diagnosis of anaerobic infections including specimen collection and culture techniques are explained. Common clinical infections associated with different anaerobic bacteria and their treatment conclude the document.
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3. • Anaerobes
Microorganisms that grow only in complete or nearly
complete absence of molecular oxygen.
Non-sporing Anaerobes – These do not form spores
and are those which usually form the Normal flora Of
Human beings and usually present in Skin, Oral cavity,
GIT, Genitourinary tract and are opportunistic in
nature.
Introduction
4. HABITAT
• Normal flora of skin, mucosal surfaces, mouth,
respiratory tract, GIT, genital tract
• Outnumber aerobes in many habitats
– mouth and skin -10 to 30 times > aerobes
– Intestines-1000 times >aerobes
• Estimated no of anaerobes in:
– Saliva – 108/ ml
– Small intestine – 105/ ml
– Colon – 1011/ gm
4
8. VIRULENCE FACTORS
Actinomyces species Including
A.israeli, A.meyeri
A.naeslundii
A.odontolyticus
Not well characerised.
Infections usually require disruptions of
protective mucosal surface of the oral cavity,
respiratory tract, GIT
Propionibacterium species
Bifidobacterium species
Eubacterium species
Mobiluncus species
No definitive virulence factors are known
Bacteroides species including
B.fragilis
B.gracilis
B.ureolyticus
Provotella species
Porphyromonas species
Fusobacterium nucleatum
These produce capsules and succinic acid,
which inhibits phagocytosis and various
enzymes that mediate cell damage
9.
10. ANAEROBIC COCCI
• Peptococcus & Peptostreptococcus – usually produce
mixed infections along with Clostridia or anaerobic
gram negative bacilli
– Puerperal sepsis & other genital infections
– Wound infections
– Gangrenous appendicitis
– UTI
– Osteomyelitis
– Abscesses in brain, lungs & other internal organs
10
12. GRAM POSITIVE BACILLI
A,israelii,A.meyeri
A.naeslundii
A.odontolyticus
species
Usually involved in mixed oral or
cervicofacial, thoracic,pelvic and
abdominal infections caused by patients
endogenous strains
Propionibacterium species Usually involved in mixed oral or
cervicofacial, thoracic, pelvic and
abdominal infections caused by patients
endogenous strains
Bifidobacterium species
Eubacterium species
Usually encountered in mixed infections of
pelvis or genito urinary tract
Mobiluncus species Usually encountered in mixed infections of
pelvis or genito urinary tract
17. PREDISPOSING FACTORS
• Trauma, Tissue necrosis, Impaired circulation, hematoma
formation or the presence of foreign bodies
• Diabetes, Malnutrition, Malignancy or prolonged
treatment with antibiotics.
• They are usually polymicrobial in nature, more than one
anaerobe can be responsible besides aerobic bacteria.
• When the infection is usually localised, it gets generalised
by hematogenous route causing Bacteremia.
18. Sequence of the events
Trauma to the sites of protective barriers allow
anaerobes of indigenous flora to gain access to
deeper tissues.
Vascular stasis
Growth & multiplication of anaerobes
Anaerobic infection
20. Polymicrobial nature of the anaerobic infection
• Contamination of the tissue by the normal flora of
the mucosa of the mouth, pharynx, GIT or genital
tract.
• Multiple species are present including other
anerobes and facultative anaerobes.
• Aerobic bacteria may also be present.
21. CHARACTERISTICS SUGGESTING ANAEROBIC
INFECTIONS
Foul smelling discharge
Infection in close proximity to mucosal surface.
Tendency to form closed space infections either as
discrete abscess (lung, brain, pleura) or by burrowing
through tissue layers
Polymicrobial nature.
Infection associated with necrotic tissue (poor blood
supply).
22. Gas formation in tissues
Failure to isolate organisms from pus (sterile pus) &
negative aerobic cultures.
Lack of response to usual antibiotic therapy.
Infection of human or animal bite wounds.
Detection of sulphur granules in pus .
Gram negative bacteraemia is more common.
26. Collection and transport of specimen
•Aspirate collection by syringe
can be sent by plugging the
needle with a cork.
•Other specimen can be
transported in robertson
cookedmeat media or in
anaerobic transport media
27. Transport of samples
• Enriched thioglycollate
broth
• Oxygen free systems
gassed with CO2 or N2
• Anaerobic pouches
28. Anaerobic specimen processing and
identification
SPECIMEN
1.Gross examination
Purulence , necrosis,
foul odour, sulphur
granules, fluoresence .
2. Gram stain
•Methanol fixed (30 sec )
smear.
• Pale , pleomorphic
filamentous Gram
negative bacteria with or
with out vacuoles ,
irregular staining
bacteriodes species.
• Pale gram negative
cocco bacilli , pigmented
prevotella species or
porphyromonas species .
3. inoculation into culture
media
• Brucella blood agar
with 5% sheep blood ,
neomycin , hemin and
vitamin k.
• Phenyl ethyl blood agar.
•Robertson cooked meat
media or enriched
thioglycolate media with
vitamin k and hemin.
29. •Thin pale gram negative
bacteria with tapering
ends(fusiform).
Fusobacterium nucleatum.
•Very small gram negative
cocci
Veillonella species.
• Kanamycin-vancomycin
laked blood agar if pale
gram negative cocco
bacilli found on gram
stain.
•Bacteroides bile esculin
agar for bacteriodes
fragils.
34. Anaerobic Jar
• Tightly sealed container in
which oxygen is completely
eliminated by hydrogen and
a catalyst.
• palladium coated aluminum
pellets acts as a catalyst.
• Methylene blue anaerobic
indicator.
• Anaerobic charge activated
with H2SO4.
35. • Colony morphology Identification
• Agar pitting Bacteroides ureolyticus
• Black or tan pigmentation Porphyromonas
• Brick red fluorescence Prevotella
• Fried egg Fusobacterium necrophorum
• Greening of medium Fusobacterium varium
• Molar tooth Actinomyces
• Speckled or bread-crumb Fusobacterium nucleatum
38. Bile Resistance
• Bacteroides resistance
to bile on bacteroides
bile esculin agar.
• Confirmatory test for
bacteroides.
39. Spot Indole test
• Growth obtained from a single,pure culture on a
blood agar plate is smeared on filter paper that
has been saturated with 1%
paradimethylaminocinnamaldehyde in 10%
concentrated HCl.
• Immediate formation of a blue color indicates
positive reaction.
43. Antibiotic resistance
• Most of bacteroides fragilis,prevotella and
porphyromonas species are resistant to
penicillins and to many cephalosporins due to
production of beta-lactamase.
• The resistance is overcome by treating with
high concentration of piperacillin,imipenem
along with beta-lactamase inhibitors.
• Bacteroides show plasmid mediated resistance
to clindamycin.
44. Treatment
• Surgical –
– Drainage of pus from abscess
– Wound debridement
– Curettage & removal of necrotic tissues
• Antibiotics –
– Metronidazole
– Penicillin
– Clindamycin
– Cephalosporins
– chloramphenicol
44
