ANAEROBIC BACTERIA

ANEROBIC BACTERIA Marker for anaerobiosis

- Lacks Superoxide Dismutase & Catalase

Collection: Needle Aspiration (NEVER SWAB)

Media:
1. Schaedler Agar

2. Lombard Dowell Agar (LD)

3. Wilkins-Chalgren broth and agar - AST media for

anerobes

4. Anaerobic PEA - gram positive anaerobes

5. THIO - enriched broth with hemin and Vit K to

enhance growth of anaerobes.
- Aerobes grow at TOP
- Strict Anaerobes grow at BOTTM
- Facultative anaerobes grows throughout
- Store at room temperature. Boil and cool before
use.
6. Bacteroides Bile Esculin Agar (BBB) - B. fragilis
- Gentamicin - inhibits AEROBES
- 20% bile - inhibits most ANAEROBES
- Laked-blood: hemolyzed by freezing and thawing
- (+) blackening due to bile and esculin
Dos and Don’ts in Anerobic Culture
- Ideal specimen is whole tissue or aspiration via needle
and syringe, remove needle ff: aspiration
- SWAB - least favorable method for collection.
- Specimens on swabs can dry out during transportation
and expose anaerobes to room air, not to be accepted
unless transported in proper anaerobic environment
- Have special nutritional requirements for HEMIN,
VITAMIN K and YEAST EXTRACT

7. Anaerobic Kanamycin Vancomycin Blood Agar

(AKV-BA) - gram negative anerobes
- Kanamycin - inhibits MOST FACULTATIVE
GRAM (-) BACILLI
- Vancomycin - inhibits to gram (+) organism and
some porphyromonas

8. Cycloserine-Cefoxitine Fructose Agar (CCFA) - for

Clostridium difficile

9. Egg yolk agar - detection of lecithinase and lipase

production by Clostridium & Fusobacterium.

Methods to Promote ANAEROBIOSIS

1. Gas Pak Jar or Mcintosh Fildes Jar, Brewer,
Torbal Jar - ANAEROBIC JARS

2. Cooked meat medium/chopped cooked meat

medium

3. Anaerobic glove box and chamber

4. Pre-reduced Anaerobically Sterilized (PRAS)

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 GRAM POSITIVE ANAEROBIC RODS GRAM NEGATIVE ANAEROBIC RODS
• Actinomyces spp. • Propionibacterium • Bacteroides • Prevotella
• Bifidobacerium • Eubacterium • Fusobacterium • Porphyromonas
• Lactobacillus • Mobiluncus
Bacteroides fragilis
Actinomyces spp. - Predominant bacteria if GIT (colon flora) - has greater
- Fungus like bacteria concentration in GIT than E. coli
- Ray fungus Branching and filamentous - Non-motile, non-hemolytic
- ACTINOMYCOSIS - disease affecting SOFT - Needs 20% bile, black on BBE (Bacteroides Bile
TISSUES Esculin), CAPSULATED, Catalase positive
- Resistant to penicillin, kanamycin, vancomycin and
Species colistin
1. Actinomyces bovis - LUMPY JAW
Fusobacterium spp.
2. Actinomyces Israeli - draining sinus tract with 1. Fusobacterium nucleatum
SULFUR GRANULES, MOLAR TOOTH - Predominant in oral cavity, may cause
COLONY (old/aged colonies), catalase negative ASPIRATION PNEUMONIA
- Colonies with “RANCID BUTTER ODOR” due
Bifidobacterium to production of Butyric Acid; Biochemically
- Highly pleomorphic rod with BIFID ends; resembles inert
bone-shaped dog biscuits - Breadcrumb colonies/Ground glass or
- Normal flora of the GIT; rare cause of pulmonary speckled appearance
infection. - Fusiform rod, spindle shape, bacilli with pointed
or tapers ends.
Lactobacillus
- Dorderlein bacillus aka Lactobacillus acidophilus 2. Fusobacterium necrophorum
- High during pregnancy - Vincent’s angina and Lemierre’s disease
- Inhibits G. vaginalis - POSITIVE FOR CHARTREUSE
- Promote C. albicans FLUORESCENCE
- Catalase negative - More virulent than F. nucleatum
- Lactobacillus casei strain Shirota (LcS) - on Yakult
Prevotella melaninogenica
Propionibacterium acnes - Bile Sensitive-Black Pigment
- Cutibacterium acnes - new name - Vancomycin Resistant, saccharolytic
- Causative agent of Acne; skin flora - Produces BRICK-RED on UVL
- Blood culture contamination
- RBC contamination Porphyromonas asaccharolytica
- Forms the so-called LOG-JAM ARRANGEMENT - Bile-Sensitive-Black pigment, red fluorescence on
- Appears as clubbed, pleomorphic rod in palisades that UVL
mimics ANAEROBIC DIPTHEROID - Vancomycin Sensitive, assacharolytic
- Catalase and indole positive
Differential test for Gram-Negative Anaerobes
Eubacterium spp.
- Pleomorphic rod usually in pairs and chains
- Normal flora of mouth and GIT
- Metabolic product: BUTYRIC ACID

Mobiluncus
- Causes Vaginitis

Differential test for GRAM POSITIVE ANAEROBIC

RODS

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 GRAM POSITIVE ANAEROBIC COCCI
1. Peptostreptococcus anaerobius - SPS sensitive -
anticoagulant for blood culture bottle, indole negative,
catalase negative and with SWEAT ODOR.
▪ Peptostreptococcus assacharolyticus - catalase
negative
2. Peptococcus - ANAEROBIC Staphylococcus; non-
hemolytic, appears in cluster, catalase (+)
▪ Peptococcus niger - Staphylococcus - like,
catalase positive.
3. Ruminococcus
4. Coprococcus

GRAM NEGATIVE ANAEROBIC COCCI

1. Veillonella parvula - fluorescence red on UVL, jaw
surgery, mouth flora, HIGHLY-SENSITIVE to
oxygen
2. Megasphera
3. Acidaaminococcus