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MICROBIOLOGY LECTURE 8 – Anaerobic Bacteria

Notes from Lecture


USTMED ’07 Sec C - AsM
• Bacteroides
Introduction to Anaerobes
o physiology & structure
Generalities  B. fragilis: most important
• gram (+) or gram (-) member
• spore forming or non-spore forming  Pleomorphic in size and shape
• cocci, bacilli, comma-shaped, spiral  Most members: grow rapidly in
• intolerance to O2, grow best at low or negative Eh
culture
• absence of: cytochrome system, superoxide  Glycolipid: has little or no
dismutase, catalase endotoxin activity – low or loss of
• normal flora: skin, mucosa, mouth, GIT pyrogenic activity
 Other species: B. ovatus, B.
Virulence Factors vulgatus
• lipopolysaccharide capsule
o Virulence Factors
• enzymes: collagenase, heparinase, lecithinase  Capsule: antiphagocytic &
• metabolic end products: fatty acids promotes abscess formation
• in some strains: endotoxin  Lipopolysaccharide: can stimulate
• infection results: when they contaminate sterile sites leukocyte migration & chemotaxis
 Agglutinins: role not known
Clinical Infections  Enzymes: hyaluronidase,
• usually insidious & may become chronic collagenase, neuraminidase, etc
• frequently produces putrid odor in infected material  O2 tolerance: capable of surviving
• gas may be present in tissue or in loculations prolonged exposure to O2
• most infections are due to moderately obligate
anaerobes o Epidemiology
• Eg. Peritonitis, pneumonitis, UTI, URTI, soft tissue  Major component of human
infections (cellulites, necrotizing fascitis), septicemia microbial flora
 B. fragilis: most commonly
Laboratory Diagnosis associated with pleuro-pulmonary,
• specimen collection: aspirated or tissue specimen intraabdominal & genital
preferred than swab infections
• sensitivity testing: should be done in life threatening  Prominent role in human disease:
infections attributed to enhanced virulence

Prereduced anaerobic plates can be o Clinical Syndromes


held in an anaerobic holding jar for a
 Cardinal features of Bacteroides
short time before and after inoculation
infections
• Endogenic
• Polymicrobic mixture
• Abscess formation

An anaerobic atmosphere can also be o Laboratory Diagnosis


created in an anaerobic pouch that hold
 Microscopy: faintly staining,
only two plates or in an anaerobic jar for
three or more plates. pleomorphic, gram (-) bacilli
 Culture: collection & transport=
O2-free system
Gas pack anaerobic jar system contains
hydrogen and CO2 generator envelope,  Biochemical ID: activity of
a disposable methylene blue indicator enzymes, metabolic by-products
and a catalyst basket in the lid  Others: resistance to: kana,
vanco, colistin
 Growth in 20% bile

o Tx, Prevention, Control


 Main approach: antibiotics with
surgical intervention
 Antibiotics:
Gas pack anaerobic pouch: bag is oxygen • B-lactam group:
impermeable and contains its own gas- cefoxitin, imipenem;
generating kit and cold catalyst • beta lactamase
inhibitors
• metronidazole
Treatment  Prophylaxis: diagnostic & surgical
• surgical debridement & resection of necrotic tissue procedures
• antibiotics: metronidazole, imipenem, clindamycin,  Endogenous spread: virtually
cefoxitin, piperacillin impossible to control

Classification Bacteroides fragilis on Brucella blood


1. Gram negative bacilli agar: non-selective medium
supplemented by Vit. K1 and hemin
• Bacteroides
• Prevotella
• Porphyromonas
• Fusobacteria
Bacteroides fragilis on Bacteroides bile
esculin agar(BBE): Bacteroides fragilis
group is resistant to bile salts; gray to
Spot Indole disk test : a plain filter paper is placed on an area of growth black colonies due to hydrolysis of
on a medium containing tryptophan. After 5 min. a drop of esculin in the medium
paradimethylaminocinnamaldehyde is placed on the disk. A greenish color
indicates the presence of indole(Fusobacterium,Propionibacterium, Growth of Bacteroides fragilis on Bacteroides fragilis in an
Porphyromonas, Prevotella, and Peptostreptococcus spp. Bacteroides bile esculin agar abscess
• History of an invasive
dental procedure or oral
trauma
 Thoracic
• Generally have a history
of aspiration with
establishment of disease
in the lungs and then
Nitrate disk test: nitrate infiltrated
spread to adjoining
filter paper is placed onto the inoculum
and after 48 hrs incubation, drops of tissue
nitrate reagents, sulfanilic acid and  Abdominal
alpha-napthylamine are added to the • Most commonly
disk. A red color indicates the presence preceded by surgery or
of nitrite trauma to the bowel
 Pelvic
• Prevotella
• Can be a secondary
o Nonspore forming
manifestation of
o May appear as slender rods or coccobacilli
abdominal actinomycosis
o Eg. P. melanogenica, P.bivia, P. disiens
or
• Could be a primary
• Porphyromonas
infection in women with
o Non-spore forming IUD
o Normal flora of mouth  CNS
• Usually represent
• Fusobacteria secondary spread form
o Appear as long, thin filaments with pointed another focus
ends o Clinical syndromes
o Colonies are hemolytic on blood agar
 Majority: cervicofacial
 Maybe acute pyogenic or slowly
evolving painless process
2. Gram positive bacilli
 Cervicofacial: swelling with
fibrosis & scarring and open
• Actinomyces
draining sinus tracts along the
• Lactobacillus angle of jaw and neck
• Propionibacterium B
 Thoracic: non-specific
• Eubacterium A
 CNS: most common manifestation:
• Bifidobacterium solitary abscess
• Rothia o Laboratory diagnosis
• Mobiluncus  Microscopy: sulphur-granules
• (CLOSTRIDIUM) (thin), gram (-) bacilli along the
periphery of the granules
Clostridia on Eggyolk agar: lecithinase-opaque white precipitate  Culture: anaerobic conditions
extending from the colony into the medium(A);  Colonies: white with domed
Lipase-iridescent sheen on the surface of colony(B) surface
Clostridium perfringens on Brucella  Biochemical media
blood agar: double zone beta o Tx, Prevention, Control
hemolysis; large colonies with peaked  Surgical debridement
centers and irregular edges after 48 hrs  Antibiotics: penicillin (DOC),
incubation
tetra, erythro, clinda
 Good oral hygiene
 Prophylactic AB: GI surgery

Sulfur granule collected from sinus


Gram stain of Clostridium spp.: gram tract in patient with actinomycosis
variable, long, thin, parallel sided,
some with swollen ends indicative of
spore formation

• Actinomyces
o Physiology & Structure
 Pleomorphic Pelvic actinomycosis
 Facultative or strict anaerobe
 Grows slowly in culture
 Typically form delicate
filamentous hyphae Molar tooth appearance of Actinomyces
 Lack mitochondria & nuclear israelii after incubation for 1 week
membrane
 Inhibited by penicillin
o Pathogenesis
 Definition: characterized by
multiple abscesses connected by
sinus tracts • Propionibacterium: Generalities
o Epidemiology o Small, gm (-) bacilli, frequently arranged in
 No evidence of person to person short chain or clumps
spread o Commonly found: skin, conjuctiva ext. ear,
 No disease originating form oropharynx, female genital tract
external source o Physiology: anaerobic or aerotolerant, non-
 All age group affected motile, catalase (+) ferment carbohydrates
 No seasonal nor occupational o 2 commonly isolated species
predilection  P. acnes
 A. israelii – most commonly - Disease caused by P. acnes
encountered o Acne: teenagers & young
o Disease classification adults
 Cervicofacial o Opportunistic infections:
• Seen in patients with patients with prosthetic
poor oral hygiene devices & IV line
- Pathophysiology
o Stimulate an inflammatory  Intermediate susceptibility to:
response • Clinda
o Production of low MW • Erythro
peptides: attract WBC to • Tetra
sebaceous follicles • Metro
- Treatment
 Specific therapy: indicated for
o Antibiotic: topical & oral
monomicrobic infections
 P. propionicus 4. Gram negative cocci
- Pathophysiology
• Veillonella, Acidaminacoccus, Megashaera
o Causes abscesses, lacrimal
• Isolated from human infections
canaliculitis, actinomycosis
• Isolates from oropharynx & colon: low degree of
• Mobiluncus virulence & represent fewer than 1% of all
anaerobic isolates
o Gram variable or gram (-) curved,
• Isolates: generally present in mixtures & clinical
nonsporing bacilli
o But classified as gram (+): signigicance difficult to assess
• Specific treatment: often not necessary
• Have gram (+) cell wall
• Lack endotoxin
• Susceptible to: vanco, clinda, erythro,
ampi
o Physiology
 Obligate anaerobes
 Fastidious, slow growing even on
enriched media
 2 species of medical importance:
• M. curtsii
• M. mulieris
 Abundant in women with bacterial
vaginosis

• Other gram (+), anaerobic bacilli


o Bifidobacterium & Eubacterium
 Commonly found in the large
intestine
o Lactobacillus
 Normal flora of urethra & female
genital tract, maintains acidic pH,
rarely cause disease
o Rothia: oropharynx

3. Gram positive cocci


• Peptostreptococcus spp.
• Peptococcus spp.
• Etc Diseases associated with Peptostreptococcus, Actinomyces,
Propionibacterium, Mobiluncus
Sodium polyanethol sulfonate(SPS) disk
test: Peptococcus anaerobius is the
only gram positive coccus that is
inhibited by SPS
-fin-
- more than 25% of all anaerobic isolates in clinical
specimens:
audsmartinez@gmail.com
ustmedc3@yahoogroups.com
• Peptostreptococcus
o Gram (+) cocci
o Normal flora: oral cavity, GIT, GUT skin
o Infections: pleuropulmonary infections,
sinusitis, brain abscesses, intraabdominal
sepsis, pelvic infections (endometritis,
pelvic abscess, salphingitis), soft tissue
infections, endocarditis, osteomyelitis
o Most infections are polymicrobial mixtures
of anaerobic & aerobic bacteria
o Only about 1% of all anaerobic bacteremias
are due to gram (+) cocci, with majority
caused by Peptostreptococci from genital
tract of women
o Bone and joint infections are usually
associated with surgical procedure
o Laboratory Diagnosis
 Complicated by 3 factors
• Contaminants
• Transport of media
should be O2-free
• Specimen should be
cultured on enriched
media for 5-7 days
o Treatment
 Usually susceptible to
• Penicillin,
• cephalosporins,
• imipenem,
• chloramphenicol

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