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    Anaerobe of Clinical Importance

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    Dayledaniel Sorveto
    This document discusses anaerobic bacteria of clinical importance. It begins by defining anaerobes and describing their habitat and potential virulence factors. It then describes the most frequently encountered genera of anaerobic bacteria, including their characteristics, associated clinical infections, and laboratory methods for diagnosis. These include spore-forming and non-spore-forming Gram-positive bacilli, Gram-negative bacilli, and anaerobic cocci. The document concludes by covering specimen collection, transport, processing, and incubation for isolation of anaerobic bacteria.

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    Anaerobe of Clinical Importance

    Uploaded by

    Dayledaniel Sorveto
    140 views43 pages
    This document discusses anaerobic bacteria of clinical importance. It begins by defining anaerobes and describing their habitat and potential virulence factors. It then describes the most frequently encountered genera of anaerobic bacteria, including their characteristics, associated clinical infections, and laboratory methods for diagnosis. These include spore-forming and non-spore-forming Gram-positive bacilli, Gram-negative bacilli, and anaerobic cocci. The document concludes by covering specimen collection, transport, processing, and incubation for isolation of anaerobic bacteria.

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    This document discusses anaerobic bacteria of clinical importance. It begins by defining anaerobes and describing their habitat and potential virulence factors. It then describes the most frequently encountered genera of anaerobic bacteria, including their characteristics, associated clinical infections, and laboratory methods for diagnosis. These include spore-forming and non-spore-forming Gram-positive bacilli, Gram-negative bacilli, and anaerobic cocci. The document concludes by covering specimen collection, transport, processing, and incubation for isolation of anaerobic bacteria.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    140 views43 pages

    Anaerobe of Clinical Importance

    Uploaded by

    Dayledaniel Sorveto
    This document discusses anaerobic bacteria of clinical importance. It begins by defining anaerobes and describing their habitat and potential virulence factors. It then describes the most frequently encountered genera of anaerobic bacteria, including their characteristics, associated clinical infections, and laboratory methods for diagnosis. These include spore-forming and non-spore-forming Gram-positive bacilli, Gram-negative bacilli, and anaerobic cocci. The document concludes by covering specimen collection, transport, processing, and incubation for isolation of anaerobic bacteria.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
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    of 43

    Anaerobe of Clinical Importance

    Important Concept
    • Anaerobe definition
    – Lack/low protective enzymes
    • superoxide dismutase/ catalase
    – Types
    • Strict/obligate
    • Aerotolerant/moderate
    Important Concept
    • Habitat
    – Exogenous anaerobes
    – Endogenous anaerobes
    • Potential virulence factors
    – Capsules, adhesins, toxins, endo/exogenous enzymes
    • MOT
    – Human/animal bites
    – traumatic puncture of oral cavity, bowel, genital tract
    – introduction of soil into wound
    Frequently encountered anaerobes

    • G (+), Spore-Forming Anaerobic Bacilli


    • G (+), Non–Spore-Forming Anaerobic Bacilli
    • Anaerobic Gram-Negative Bacilli
    • Anaerobic Cocci
    G (+), Spore-Forming Anaerobic Bacilli

    • Clostridium
    – Terminal, subterminal, central
    G (+), Spore-Forming Anaerobic Bacilli
    • Clostridium perfringes
    – Virulence
    • enterotoxins = disrupts
    mucosal cells
    • α-toxin (lecithinase), β-toxin =
    destruction of host cells
    – Clinical infections
    • Food poisoning (ingestion)
    – type A = mild and self-limiting
    – type C (enteritis necroticans) =
    rare and severe
    • Bacteremia
    • Gas gangrene
    G (+), Spore-Forming Anaerobic Bacilli

    • Clostridium perfringes
    – Lab diagnosis
    • Gram variable
    – Subterminal spores
    – Large boxcar shape
    • Culture
    – double zone of hemolysis
    – EYA: (+) lecithinase, (-) lipase
    – Nagler plate
    – Reverse CAMP test
    G (+), Spore-Forming Anaerobic Bacilli

    • Clostridium perfringes
    – Lab diagnosis
    • Gram variable
    – Subterminal spores
    – Large boxcar shape
    • Culture
    – double zone of hemolysis
    – EYA: (+) lecithinase, (-) lipase
    – Nagler plate
    – Reverse CAMP test
    G (+), Spore-Forming Anaerobic Bacilli

    • Clostridium botulinum
    – Virulence
    • botulinum toxins type A,B
    and E
    – potent neurotoxin; flaccid
    paralysis
    – potential boiterrorist agent
    – Clinical infection
    • Foodborne botulism
    • Wound botulism
    • Infant botulism
    G (+), Spore-Forming Anaerobic Bacilli

    • Clostridium botulinum
    – Lab diagnosis
    • Microscopic exam.
    – Straight w/ blunt ends
    – subterminal spores (tennis
    racket)
    • Culture
    – beta hemolytic
    G (+), Spore-Forming Anaerobic Bacilli

    • Clostridium tetani
    – Virulence
    • Neurotoxin
    tetanospasmin
    – spastic paralysis
    – Clinical infection
    • Tetanus
    • Lock jaw (trismus)
    • risus sardonicus
    G (+), Spore-Forming Anaerobic Bacilli

    • Clostridium tetani
    – Virulence
    • Neurotoxin
    tetanospasmin
    – spastic paralysis
    – Clinical infection
    • Tetanus
    • Lock jaw (trismus)
    • risus sardonicus
    G (+), Spore-Forming Anaerobic Bacilli

    • Clostridium tetani
    – Lab diagnosis
    • Microscopic exam.
    – straight w/ blunt ends
    – round terminal spores
    (drumstick/lollipop/tack
    head)
    • Culture
    – narrow zone of beta
    hemolysis (may swarm)
    G (+), Spore-Forming Anaerobic Bacilli

    • Clostridium difficile
    – Virulence
    • Enterotoxin (Toxin A)
    • Cytotoxin (toxin B)
    – Clinical infection
    • nosocomial infection
    • pseudomembranous
    colitis
    G (+), Spore-Forming Anaerobic Bacilli

    • Clostridium difficile
    – Lab diagnosis
    • Microscopic exam.
    – straight w/ oval
    subterminal spore
    • Culture
    – Non hemolytic, horse
    stable odor, yellow
    green fluorescent
    – Yellow ground glass in
    CCFA (indicator –
    neutral red)
    G (+), Spore-Forming Anaerobic Bacilli

    • Clostridium septicum
    – Clinical infection
    • Malignancy in the GI tract
    – Lab diagnosis
    • gram variable
    – Oval subterminal spore
    • Culture
    – “Medusa head”-like
    appearance
    – beta hemolytic, swarming
    G (+), Spore-Forming Anaerobic Bacilli

    MOTILITY LECITHINASE LIPASE LACTOSE GLUCOSE


    C. perfringens - + - + +
    C. botulinum + - + - +
    C. tetani + - - - -
    C. difficile + - - - +
    Frequently encountered anaerobes
    • G (+), Spore-Forming Anaerobic Bacilli
    • G (+), Non–Spore-Forming Anaerobic Bacilli
    • Anaerobic Gram-Negative Bacilli
    • Anaerobic Cocci
    G (+), Non–Spore-Forming Anaerobic Bacilli

    • Actinomyces israelli
    – Clinical infection
    • Actinomycosis
    – Lab diagnosis
    • branching filamentous
    rods
    • Culture
    – Molar tooth
    appearance
    G (+), Non–Spore-Forming Anaerobic Bacilli

    • Propionibacterium
    acnes
    – Clinical infection
    • Acne
    • contaminant of blood
    culture
    – Lab diagnosis
    • pleomorphic diptheroid-
    like rod
    • “Picket fences” or
    “Anaerobic diphtheroids”
    G (+), Non–Spore-Forming Anaerobic Bacilli

    • Bifidobacterium
    spp.
    – Clinical infection
    • pelvis, GUT,
    abdominal infection
    – Lab diagnosis
    • Rods w/ bifurcated
    ends (dog bones)
    G (+), Non–Spore-Forming Anaerobic Bacilli

    • Lactobacillus acidophilus
    – Boas-oppler bacillus: GI tract
    – Doderlein bacilli : vaginal canal
    – Clinical infection
    • Bacterial vaginosis
    • Bacteremia, endocarditis
    – Lab diagnosis
    • Tomato juice agar
    Frequently encountered anaerobes

    • G (+), Spore-Forming Anaerobic Bacilli


    • G (+), Non–Spore-Forming Anaerobic Bacilli
    • Anaerobic Gram-Negative Bacilli
    • Anaerobic Cocci
    Anaerobic Gram-Negative Bacilli
    • Clinical infections
    – associated with peritoneal infections following
    disruption of the GI lining.
    – Brain abscesses
    – mixed biota in diabetic foot ulcers
    – Hallmark: foul odor
    – Virulence: capsules, endotoxins, succinic acids,
    enzymes
    Anaerobic Gram-Negative Bacilli
    • Bacteroides fragilis
    – Lab diagnosis
    • GS: Bipolar staining,
    pleomorphic
    • growth in 20% bile, KVLB
    agar, BBE agar
    • Resistant to vancomycin,
    kanamycin, colistin
    • (+) esculin hydrolysis,
    catalase
    • Saccharolytic
    Anaerobic Gram-Negative Bacilli
    • Prevotella spp.
    – Lab diagnosis
    • Growth on KVLB agar =
    Protoporphyrin (dark
    pigment)
    • Resistant to
    vancomycin, kanamycin
    • Inhibited in 20% bile
    • Brick red under UV light
    • Saccharolytic
    Anaerobic Gram-Negative Bacilli
    • Porphyromonas
    – Lab diagnosis
    • more mucoid than Prevotella
    • Brick red under UV light
    • Susceptible to vancomycin
    • Assacharolytic
    • inhibited by 20% bile
    Anaerobic Gram-Negative Bacilli
    • Fusobacterium nucleatum
    – Lab diagnosis
    • long, thin, tapered rods
    • Resistant to vancomycin
    • Susceptible to colistin,
    kanamycin
    • No growth on KVLB
    • Brucella agar:
    – Bread crumblike, Speckled,
    Smooth
    – Greening on exposure to air
    – Fluoresces chartreuse
    – Foul odor
    Frequently encountered anaerobes

    • G (+), Spore-Forming Anaerobic Bacilli


    • G (+), Non–Spore-Forming Anaerobic Bacilli
    • Anaerobic Gram-Negative Bacilli
    • Anaerobic Cocci
    Anaerobic cocci
    • Peptostreptococcus = G (+)
    • Veillonella = G (-)

    – Clinical infection
    • Brain abscess, gingivitis, other periodontal diseases
    – Lab diagnosis
    • Brick red under UV light
    Lab diagnosis
    Specimen Collection and Transport
    • Aspirates
    – PRAS (prereduced
    anaerobically sterilized)
    • Swabs
    – 0.5 ml sterile THIO
    • Tissue biopsy
    – anaerobic pouch
    • Blood
    • Fecal samples
    Specimen Collection and Transport
    • Rubber stoppered vials
    • Oxygen free vials
    • Anaerobic pouch
    Specimen processing
    • Macroscopic examination
    – foul odor
    • Fusobacterium, Porphyromonas, Prevotella,
    Bacteroides
    – brick red under UV light
    • Porphyromonas, Prevotella
    – black exudate/necrotic tissue
    • Porphyromonas, Prevotella
    – sulfur granules
    • Actinomyces, Propionibacterium
    Specimen processing
    • Microscopic examination
    – Presumptive id
    – reveals the presence of leukocytes, SEC
    – serve as a valuable quality control tool
    Specimen processing
    • Inoculation of appropriate plated/tubed
    media
    – special nutritional requirements
    • vitamin K, hemin, and yeast extract
    Specimen processing
    • Inoculation of appropriate plated/tubed
    media
    – Anaerobic blood and Brucella blood agar
    • Nonselective for facultative and strict anaerobes
    – PEA and CNA Blood Agar
    • G (+) strict/facultative anaerobe
    – Thioglycolate Broth, cooked meat broth
    Specimen processing
    • Inoculation of appropriate plated/tubed
    media
    – Kanamycin-vancomycin-laked blood agar
    (KVLB)
    • Selective for Bacteroides, Prevotella
    • Kanamycin = inhibits F. gram (-) bacilli
    • Vancomycin = inhibits F. and strict gram (+) bacteria
    – Bacteroides bile esculin agar (BBE)
    • Sel/Diff. for B. fragilis
    Specimen processing
    • Inoculation of appropriate plated/tubed
    media
    – Cycloserine cefoxitin fructose agar (CCFA)
    • Sel/Diff. for C. difficile (golden yellow under UV light)
    – Egg-yolk Agar (EYA)
    • determination of lipase/lecithinase production
    Specimen processing
    • Other tests
    – Antibiotic disk test
    • vancomycin (5 ug.),
    colistin (10 ug.),
    kanamycin (1mg.)
    • resistant: <10mm;
    susceptible: >/=10mm
    ZOI
    Specimen processing
    • Other tests
    – Susceptibility test in SPS
    • susceptible: >12mm
    – Catalase
    – Nitrate
    – Indole
    – Growth in 20% bile
    Specimen processing
    • Incubation
    – Anaerobic jars
    • Gas generator : Gas pak
    • Catalyst : palladium pellets
    • Dessicant : silica gel (blue to pink)
    • Redox indicator : methylene blue, resazurin
    •Thank you for
    listening! :)

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