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Anaerobes
Stack #159197
Question | Answer |
---|---|
Obligate aerobe | 5-20% O2 |
Microaerophile | 5% O2 |
Faculative anaerobe | Presence or absence of O2 |
Aerotolerant anaerobe | <5% O2 |
Obligate anaerobe | Can't tolerate O2 |
Capnophile | 5-10% CO2 |
Anaerobes in respiratory tract | F. Nucleatum; Porphyromonas spp; Actinomyces, Peptostreptococcus, Campylobacter, Prevotella |
Anaerobes of the skin | Propionibacterium acnes; Eubacterium spp; Peptostreptococcus |
Anaerobes of the genitourinary tract | Lactobacillus; Peptostreptococcus; Bifidobacterium; Fusobacterium; Prevotella; Veillonella; Mobiluncas |
Anaerobes of the gastro tract | Bifidobacterium; Eubacterium; Clostridium; Peptostreptococcus; Bacteroides fragilis group; Campylobacter; Fusobacterium; Porphyromonas; Prevotella; Sutterella; Veillonella; Biophila |
Two ways for a patient to develop an anaerobic infection. | 1. Trauma of skin or mucous membrane lining allowing bacteria entrance 2. Tissue w/ less O2 due to impaired blood flow; necrotic |
7 clues that an infectious process involves anaerobes when examining a specimen. | 1. Infection is in close proximity to a mucosal surface 2. Infection persists despite aminoglycoside therapy 3. Presence of foul odor 4. Gas bubbles 5. Presence of sulfur granules 6. Black or brick red fluorescence 7. Gram stain morphology |
Unacceptable specimens for anaerobic culture: | Stool, exudates from abscess collected by swab; voided midstream urine, vaginal swab |
Acceptable specimens for anaerobic culture: | Anything collected by aspiration; Blood; Aspirated abscess material; cerebrospinal fluid |
How to collect and handle aspirates. | Needle & syringe; be sure to expel all air in syringes, transfer into sterile O2 & O2 free tubes. |
How to collect and handle swabs. | Most wound cultures. O2 free swab for anaerobes; O2 free transport container w/ gel tube |
How to collect and handle tissue. | O2 free transport tube, be sure to keep it upright |
How to collect and handle blood | Blood culture bottles, anaerobic & aerobic. syringe - transfer to anaerobic bottle first; vent aerobic |
Specimen integrity(container, time of collection) | Appropriate specimen, transport device, age of specimen, dried out? |
Specimen appearance | Foul odor, necrotic tissue or exudate black, sulfur granules, bloody, purulent |
Fluorescent brick-red w/ Wood's lamp | Porphyromonas & Prevotella |
4 reasons to examine gram-stained smear of specimen | 1. Reveal types & general numbers of bacteria 2. reveal the presence of leukocytes 3. may provide a presumptive id 4. quality control technique |
Media: Anaerobes have requirement for (3) | Vitamin K, hemin & yeast extract |
What is PRAS media? Why is it preferred? | Pre-reduced anaerobically sterilized media. It is the best anaerobic media. Boil media to remove O2, autoclave & replace air w/ O2 free gas mixture. |
5 anaerobic medias are: | Anaerobe blood agar (Brucella blood agar); Bacteroides Bile esculin (BBE); KVLB agar; PEA agar; Anaerobe broth (thio) |
Anaerobic/Brucella Blood Agar | Nonselective/enriched. All obligate & faculative anaerobes. Sheep's blood, Vit K, & hemin |
Media: Vit K - required by ______ | Porphyromonas spp. |
Media: hemin - enhances growth of _______ | Bacteroides spp. |
Bacteroides Bile Esculin Agar Plate grows: | Primarily for B. fragilis. Supports growth of bile-tolerant Bacteroides spp; but, some Fusobacterium mortiferum, Klebsiella pnuemoniae, enterococci & yeast may grow limitedly. |
Bacteroides Bile Esculin Agar Plate consists of: | Selective; contains gentomicin, 20% bile, and esculin. B. fragilis will turn the normally lt. yellow medium to brown do to hydrolysis. |
Gentomicin | Inhibits most aerobic organisms |
20% Bile | Inhibits most anaerobes |
Kanamycin-Vancomycin Laked Blood (KVLB) Grows: | Grows Prevotella & Bacteroides spp. but yeasts & kanamycin-resistant, faculative g- bacilli will also grow |
KVLB consists of: | Kanamycin, vancomycin, & lakes blood. |
Kanamycin | Inhibits most faculative gram neg. bacilli |
Vancomycin | Inhibits most g+ organisms & vancomycin-sensitive strains of Porphyromonas spp. |
Laked Blood | Accelerates production of brown-black pigmented colonies by certain Prevotella spp. |
PEA agar (Phenylethyl alcohol) | Grows all obligate anaerobes & g+ faculative anaerobes. Selective & contains PEA. |
Pheynylethyl alcohol | Supresses growth of any faculative g- bacilli; esp swarming Proteus spp. |
Anaerobic broth (thio) grows: | Virtually all bacteria. Obligate aerobes & microaerophiles near top, obligate anaerobes at the bottom & faculative anaerobes throughout. |
Anaerobic broth (thio) used for: | backup source of culture material as obligate anaerobes can be easily overgrown and killed by faculative organisms present |
Glove box | Anaerobic chamber with gloves built in (no O2 at all) |
Gloveless anaerobic chamber | Lets a little O2 in |
Anaerobic jars | Expose bacteria to O2 while working on them. Easily replaceable/handlable |
Anaerobic bags & pouches | Like a ziplock, contains same as jars. Catalist (alluminum pellets to remove O2), desicant, indicator, anaerobic gas |
Catalyst (formula) | H2 + CO2 --> H2O + CO2 20-25minutes |
Indicator | Rizazorin (pink) or Methylene blue (blue) |
Organisms with foul odor | C. difficile, Fusobacterium, Porphyromonas |
Double zone of hemolysis | Clostridium perfringens |
Presumptive ID | Colony morphology, gram-stain micro. morphology, results of several rapid tests |
Definitive ID | Commercially available biochemical or enzyme based ID systems. Ex: Rapid ANA - enzyme based, 4 hrs, most clinically significant anaerobes |
Clostridium tetani | G+ rod, spore forming. Transmission soil contaminates sharp objects - spores enter wound and germinate. Disease produces neurotoxin, inhibits release of neurotransmitters leading to muscle spasm & difficulty breathing. Lock-jaw(trismus), risus sardonicis. |
Clostridium botulinum | G+ rod, spore forming. Foodborne - ingest preformed toxin, prevents release of ATC leading to paralysis & death or spores introduced into wound. Infant botulism - spores ingested, colonize in GI tract & release toxin. Produces neurotoxin. Antitoxin & ant |
Clostridium difficile | G+ rod spore forming. Antibiotic assoc. pseudomembranous colitis. Transmission: part of normal intestinal flora, nonsocomial. Toxin: Toxin A (enterotoxin) affects metabolism of intestinal epithelium, loss of electrolyes & fluids. Toxin B (cytotoxin), acts |
Clostridium perfringens | G+ rod spore forming. Myonecrosis (gas gangrene): organisms contaminate wounds, release exotoxins causing necrosis & spread infection into deeper tissue. GI tract, can lead to shock/fatality. Foodborne disease - usually mild. Treatment antibiotics, remove |
Micro morphology of Clostridia tetaniC. botulinumC. perfringens | G+ rods terminal spores, drumstick shapeG+ rods terminal spores, tennis racket shapeG+ rods box-car shaped, spores difficult to detect |
Egg Yolk agar | Lecitinase: white zone inside the agarLipase: mother of pearl/gas on water - white sheen on top of colonies |
Cycloserine-cefoxitin-fructose agar (CCFA) | Selective/differential agar for C. difficile; yellow, ground glass colonies |
Actinomyces israelii |