Anaerobic bacteria

Ni Nengah Dwi Fatmawati

Department of Clinical Microbiology
Faculty of Medicine Udayana University/
Sanglah General Hospital

Classification of bacteria based on

oxygen requirement
1. Obligate/ strict aerobic bacteria: at the top of the test tube in
order to absorb maximal amount of oxygen that Is needed
for metabolism (15-21% oxygen)
2. Capnophilic bacteria needs 5-10% of CO2 for their growth.
3. Obligate or strict anaerobic bacteria gather at the bottom to
avoid oxygen (0% oxygen)
4. Facultative bacteria: mostly at the top, since aerobic
respiration is advantageous; but as lack of oxygen does not
hurt them, they can be found all along the test tube.
5. Microaerophiles gather at the upper part of the test tube but
not at the top. They require oxygen, but at a lower
concentration (5% oxygen)
6. Aerotolerant bacteria are not affected at all by oxygen, and
they are evenly spread along the test tube.

Anaerobic bacteria
Definition:
Bacteria that is able to replicate in the
absence of oxygen
Oxygen is toxic to anaerobic bacteria
After celullar metabolism, there is
production of superoxide anion (O2-) and
hydrogen peroxide (H2O2). Also OH-

4O2- + 4H+
2H2O2

Superoxide
dismutase
Catalase

2H2O2+ O2
2H2O+ O2

Sources of Anaerobic Infections

Endogenous microbiota (Actinomyces,
Clostridium difficile, Bacteroides spp, etc)
Intestinal anaerobes
Oral anaerobes

Exogenous
Clostridium tetani (tetanus)
Clostridium botulinum (botulism)

Spores
Spore positioning and species
Terminal spore
Central spore

C. tetani
C. perfringens

Subterminal spore C. septicum, novyi, histolyticum

difficile, botulinum

Indications of Involvement of
Anaerobes in Infectious Processes
Infections is in close proximity to a mucosal
surface
Presence of foul odor
Presence of large quantity of gas
Presence of black colour or brick-red
fluorescence
Presence of sulfure-granules
Distinct morphologic characteristics in Gramstained preparation

Predisposing Factors
Human or animal bite wounds
Aspiration of oral contents into the lungs after vomiting
Tooth extraction, oral surgery, traumatic puncture of the
oral cavity
Gastrointestinal tract surgery or traumatic puncture of
the oral cavity
GI tract surgery or traumatic puncture of the bowel
Genital tract surgery or traumatic puncture of the genital
tract
Introduction of soil into a wound

Potential pathogens by anatomic site

Oral cavity
Saliva
Fusobacterium nucleatum
Prevotella melaninogenica
Prevotella oralis group
Bacteroides ureolyticus group
Peptostreptococcus spp.
GI tract
Terminal ileum and colon: Bacteroides spp.
(principally members of the B. fragilis group)
Prevotella spp.

Clostridium spp.
Peptostreptococcus spp.
Female genital tract
Peptostreptococcus spp.
Bacteroides spp.
Prevotella bivia

3 critical steps in diagnosis of

anaerobic infection
1. Proper specimen collection
Specimens must be collected properly for avoiding the
contamination by the normal flora.
Air must be expelled from the syringe used to aspirate the
abscess cavity, and the needle must be capped with a sterile rubber
stopper.
Examples of specimens unacceptable for anaerobic culture
Sputum collected by expectoration or nasal tracheal suction
Bronchoscopy specimens
Culture of protected-brush specimens or bronchoalveolar lavage
fluid obtained by bronchoscopy is controversial.
Samples collected directly through the vaginal vault
Urine collected by voiding
Feces

3 critical steps in diagnosis of

anaerobic infection
2. Rapid transport of specimens to the microbiology
laboratory, preferably in anaerobic transport media
3. Proper handling of specimens by the laboratory

Laboratory diagnosis
To cultivate anaerobes, the specimen should not be
exposed to air

Sources undetermined

Acceptable Specimens
for Anaerobic Bacteriology
Anatomic Source

Specimens and Recommended

Methods of Collection

CNS

Cerebrospinal fluid, aspirated abcess

material, tissue from biopsy or autopsy

Localized abcess

Needle and syringe aspiration of closed

abcesses

Decubitus ulcers

Aspirated pus

Draining wounds or sinus tract

Aspirated material

Deep tissue or bone

Specimens obtained during surgery

from depths of wound or underlying
bone lesion

Intra abdominal

Aspirate from abcess, ascites fluid,

peritoneal fluid, tissue

Urinary

Suprapubic bladder aspiration

Other

Bone marrow, synovial fluid, bipsied

tissue from any normally sterile site

Unacceptable Specimens
for Anaerobic Bacteriology
Throat swabs, nasopharyngeal swabs, sputum
from NF swab or ET suction, bronchial washing,
expectorated sputum
Large bowel contents, rectal swabs, gastric and
small bowel contents
Voided or catheterized urine
Vaginal, cervical or urethral swab
Surface swab from decubitus ulcers

Anaerobic bacteria
Gram positive, Spore-forming Anaerobic Bacilli
Clostridium spp. (C. perfringens, C. botulinum, C.
difficile, C. tetani )

Gram positive, Non spore-forming anaerobic Bacilli

Actinomyces israelli

Anaerobic Gram negative Bacilli

Bacteroides fragilis, Prevotella, Porphyromonas,
Fusobacterium

Anaerobic Cocci
Peptostreptococcus, Veilonella

Clostridium species
Gram-positive, spore-forming bacilli
Sporulation occurs in nutrient-limiting conditions
Spores are resistant to extremes of dryness,
heat (boiling), and many chemical disinfectants
They can persist in the environment for weeks to
months and germinate deep in the soil where
the conditions are anaerobic
In favorable conditions (e.g., in human tissues),
spores germinate and may produce toxins

Clostridium perfringens
Spore-forming
(central)
Gram positive

Minor

Major

C. perfringens Virulence Factors

C. perfringens Nagler Reaction

NOTE:

Lecithinase (-toxin; phospholipase) hydrolyzes

phospholipids in egg-yolk agar around streak on right.
Antibody against -toxin inhibits activity around left
streak.

C. perfringens Double Hemolysis

Clostridium perfringens
Soft tissue infections (cellulitis, myositis
myonecrosis)
Food poisoning
Septicemia

Diagnosis: Gram stain (spore forming

Gram positive bacilli), culture (fast grower)
Treatment, Prevention, Control
Surgical debridement
Antibiotics
Proper wound care

C. tetani

C. tetani

C. botulinum

Mechanism of Action of
Botulinum Toxin

C. botulinum

C. difficile
Physiology and Structure
Gram positive, spore-forming bacillus
Strict anaerob

Epidemiology
Ubiquitous
Colonizes intestines
Overuse of antibiotic induces overgrowth of
this organism
Could be an exogenous source of infection

C. difficile Virulence Factors

C. difficile

Gram Positive Non spore forming

Anaerobic Bacilli
Actinomyces, Bifidobacterium,
Mobiluncus, Propionibacterium
Clinical infections:
Actinomycosis: Actinomyces israelii
Actinomycosis: chronic, granulomatous
with development of sinus tract and fistulae
Gram stain: non-spore forming, thin, grampositive bacilli
Bacterial vaginosis: Mobiluncus spp with other
anaerobic bacteria ( Prevotella, Gardnerella vaginalis)

Anaerobic Gram Negative Bacilli

Members of endogenous protein (oral cavity
and GI tract bacteria)
B. fragilis group, Porphyromonas, Prevotella,
and Fusobacterium
Found in mixed infections (brain abscess)
Porphyromonas, Prevotella, and
Fusobacterium
From blood cultures or peritoneal infections
B. fragilis

Anaerobic Bacteria that Harmless

for Human Body
Lactobacillus spp.
Gram-positive pleomorphic bacilli
(coccoid, spiral shaped)
Anaerobic (aerotolerant anaerob)
Normal flora in human mouth, GI tract,
female genital tract
Lactobacilli produce lactic acid (from
glycogen) reduce pH and suppresses
overgrowth of Bacterial vaginosiscausing bacteria

Anaerobic Bacteria
Anaerobic bacteria do not require
oxygen for their growth
Harmless bacteria (normal flora)
Harmful bacteria
Consideration to recognize anaerobic
bacterial infection
Consideration for samples collection
and preservation to isolate anaerobic
bacteria

Thank you