ANAEROBIC

BACTERIA/dra.sison/nie2013

CHARACTERISTICS
o
No enzymes for rotection against toxic effect of molecular O
o
Require envt w/ low oxidation-reduction potential
o
Lack cytochrome which use O metabolism
o
Lack catalase, superoxide dismutase & peroxisdase
CLASSIFICATION
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Obligate

Lack superoxide dismutase and catalase

Susceptible to lethal effects of O

o
Facultative/Aerobe

Grow better under anaerobic condition than under aerobic

Can rapidly consume all available O & change to aerobic

metab once a facultative anaerobe enter site of infection thus
allowing anaerobic bacteria that are present to grow & produce
dse
o
Aerotolerant

Can grow in the presence of O atmospheric level(poorly)

o
Microaerophilic

Require low levels of O for growth (1-3% p O)

Cant tolerate the level present in the atmosphere

ANATOMICAL SITES FOR ENDOGENEOUS ANAEROBE

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Mucosal surfaces: high concentration in mouth & GIT
o
Respiratory Tract: 90% of anerobic bacteria present in mouth

Bad breath

Aspiration pneumonia if (+) infection in oral cavity

Skin is frequently the source of contaminated bld culture specimen

o
GUT: Rarely cause infection

Cervical & Vaginal areas have 50% anaerobe

o
GIT: 2/3 of all bacteria are in the stool, only cultured anaerobically;
Clostridium difficile suspected

FACTORS PREDISPOSE PATIENTS FOR ANAEROBIC INFECTION

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Trauma to mucosal membranes or skin
o
Interruption of blood flow
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Tissue necrosis
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Decrease in redox potential in tissues

o
o

o
o

Aspirates

Tissue

Blood

Not acceptable d/t possibility of contamination of anaerobic

microbes w/ the microbiota presence on the skin
Collected w/ needle & syringe
Excess air expressed from syringe
Specimen injected into O free transport tube or vial
Placed in an O free transport bag or vial
Aerobic & anaerobic bottles are collected for blood culture

PROCESSING CLINICAL SAMPLE FOR CULTURE

Placed in an anaerobic chamber or holding device while awaiting processing
Procedures

Macroscopic exam of specimen

Gram stain (methanol fixation instead of heating)

Inoculation in an anaerobic media

Anaerobic incubation

ESTABLISHING OF ANAEROBIC CONDITION

o Candle Jar microaerophilic, facultative

Candle flame will consume most of the O in the jar

Produce an elevated level of CO

These
o Anaerobic jar w/ atmosphere generator (foil envelopes) release hydrogen
& CO + catalyzer ( should be regenerated at regular interval) after
addition of water

Consist of polycarbonate jar, a lid w/ a gasket to prevent

air flow, a strip infused w/ methylene blue. & a pouch w/
Na borohydride, Na bicarbonate, citric acid & a
palladium catalyst
o Anaerostat bottle w/ anaerobic atmosphere
o Vinyl Anaerobic Chambers provide a strict anaerobic atmosphere using a
palladium catalyst & hydrogen gas to produce 5% CO

ANAEROBIC INCUBATION
o Anaerobic chambers
o Anaerobic bags or pouches
o Anaerobic jars

Gas-Pak envelops generate CO & H w/c combines

w/ O

H is explosive, palladium catalyst must be

used

CLINICAL MANIFESTATION OF AEROBIC INFECTION

o
Purulent discharge
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Close proximity to a mucosal surface
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Infection persist despite antibiotic therapy
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Foul/Putrid odor
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Presence of large quantities of gas (bubbling or cracking sound when tissue is
palpated)
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Black color or brick red fluorescence
o
Sulfur granules
INFECTIONS PRODUCED
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Brain abcess
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Oropharyngeal, Pleuropulmonary Inraabdominal, Female genital tract i
o
Bacteremia

o
Endocarditis
o
Skin, soft tissue & bone infection

TYPICAL ANAEROBIC MEDIA

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Anaerobic blood agar (BRU/BA)
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Bacteroides bile esculin agar (BBE)
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Kanamycin-vancomycin-laked blood agar (KVLB) permits
gowth of facultative anaerobes
o
Phenyl-ethyl alcohol agar (PEA)
o
Anaerobic broth like thioglycollate (THIO) or chopped meat
METHODS FOR EXCLUDING OXYGEN
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Fluid Thioglycollate Media (FTM) supports aerobic & anaerobic
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Sodium Thioglycollate removes O from the medium
o
Resazurin as an oxidation reduction indicator dye that turns pink in
presence of O

Anaerobic jar

SPECIMEN FOR ANAEROBIC CULTURE

o
Swabs

Anaerobic

Aerobic
incubator
-

ANAEROBIC
BACTERIA/dra.sison/nie2013
Facultatively
anerobic
Aerobic

Foul odor when opending anaerobic jar

Colonies are present on the anaerobic incubated media but
non on aerobic media
Good growth on BBE
Coloneis oon KVLB that are pigmented of fluorescent
Double zone of hemolysis on blood agar

PRESUMPTIVE IDENTIFICATION OF ANAEROBES

o
Aerolorence; Fluorescence
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Special-potency antimicrobial disks
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Catalase test; Spot indole Test; Motility test
o
Lecithinase & Lipase reactions
o
Presumpto Plates ang dame ayoko na

FREQUENTLY ENCOUNTERED ANAEROBES

Gram-positive spore forming anaerobic bacilli

Clostridium
most from exogenous sources
C. diffcile is most often detected via direct stool antigen
ex: tetanus, gas gangrene, botulism, food poisoning

Cocci

Anaerobic cocci endogenous

a) Peptostreptococcus gram positive
b) Veilonella spp gram negative

GUT (lumina)
Urethra (distal): Gram (-) bacilli
o
Bacteroides spp

Gram (-)
Bacteroides
Prevotella
Porphyromonas
Fusobacteria

Veilonella

Gram (+)
Actinomyces
Lactobacillus
Propronibacterium
Eubacterium, bifidobacterium,
Arachnia
Clostridium
Peptostreptococcus

PATHOGENESIS

Prevotella bivia & Prevotella disiens dominate

among gram (-) rods

Skin (sebaceous glands)

Propionibacterium spp acne genesis; infect plastic shunt
Peptostreptococci spp
Indigenous anaerobes play a role in maintaining health
Anaerobes provide a natural barrier to prevent colonization of unwanted
organisms including pathogens
W/in the GIT they provide a source of vitamins, fatty acids & cofactors
They degrade toxic or potentially cancer producing compounds
Role in maturation of immune system of neonates through stimulation of
antibody

Bacill
i

Anaerobic gram-negative bacilli

endogenous
a) Bacteroides fragilis
b) Porphyromonas spp
c) Prevotella spp
d) Fusobacterium spp

ANAEROBIC COMMENSAL FLORA OF MAN

Intestine (lumina) major site: 95-99% total bacterial mass
Bacteroides spp
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most common genus of normal indigenous
anaerobic flora
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Bacteroides thetaioitaomicron -most prevalent
species found

o
Prevotella soo
o
Fusobacterium spp
Female genital tract
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Lactobacillus low ph
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Peptostreptococci spp
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Clostridium perfringens
o
Non sporeforming bacilli & gm (-)
o

Infections are due to combination of bacteria that function in synergistic

pathogenecity
B fragilis
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single most important pathogen among anaerobes
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Virulence: capsular polysaccharide intraabdominal abcess
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Produces superoxide dismutase
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Can survive in presence of oxygen for days
o
Clindamycin & Metronidazole for treatment
Some produce collagenase & heparinase damage tissue
Lipopolysaccharide endotoxic activity

GOD BLESS US ALL!!