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Jarreau Review
For m(ASCP) Exam
| Question | Answer |
|---|---|
| Temperature for Psychrophiles | cold-loving 15 C |
| Temperature for Mesophiles | moderate 37 C (most pathogenic orgs) |
| Temperature for Thermophiles | heat loving 50-60 C |
| pH for most Bacteria | 6.5 - 7.5 |
| pH for most Fungi | 5.0 - 6.0 |
| O2 needs of Aerobes | require O2 |
| O2 needs of Facultative Anaerobes | can grow w/ or w/out O2 |
| O2 needs of Obligate Anaerobes | harmed by O2 |
| Aerobes & Facultative Anaerobes possess [these] for neutralizing toxic superoxide radicals | catalase, peroxidase, superoxide dismutase |
| atmospheric needs of Microaerophiles | prefer lower O2 than air |
| atmospheric needs of Capnophiles | prefer higher CO2 than air |
| autoclave sterilizes how? QC org? | 15 psi, 121 C, 15 min; Bacillus stearothermophilus |
| hot air oven sterilizes how? | 170 C, 2 hrs |
| pasteurization inhibits (not sterilizes) how? | 140 C, 3 sec |
| filtration pore size .22 micron - .45 micron used for what? | sugar solutions, urea media, vaccines |
| what's the deal with dessication? | no multiplication, orgs stay viable (lyophillization) |
| osmotic pressure in hypertonic solution sterilizes how? Used for what? | plasmolysis; cured meat, fruit preserves |
| radiation sterilizes how? | forms hydroxyl radicals; damages DNA |
| phenol disinfects how? | damages cytoplasmic membrane, denatures protein |
| halogens (iodine & chlorine) disinfect how? | oxidize |
| alcohols disinfect how? | denature protein, dissolve lipids |
| difference between sterilization & disinfection | sterilization: kills all microorgs including spores & viruses; disinfection: inactivates/inhibits, not necessarily spores |
| Beta-lactam Action & uses penicillins, cephalosporins, carbapenams (imipenam), monobactams (azotreonam), beta-lactamase inhibiting combinations (augmentin) | inhibits cell wall synthesis; oxacillin, nafcillin, methicillin for penicillin-resistant Staph |
| Beta-lactam examples | |
| glycopeptides action & uses vancomycin | inhibits cell wall synthesis; C.diff & MRSA |
| aminoglycosides action & uses gentamicin, tobramycin, amikacin | inhibits protein synthesis, 30S subunit; not against anaerobes, use w/ penicillin for Enterococcus |
| tetracyclines action & caution tetracycline, doxycycline | inhibits protein synthsis, 30S subunit; affects bone & teeth in children; may lead to superinfection of yeast |
| chloramphenicol action & caution | inhibits protein synthesis, 50S subunit; can cause aplastic anemia |
| macrolides action & uses erythromycin, clindamycin | inhibits protein synthesis, 50S subunit; clin for GP & GN anaerobes |
| quinolines action & uses ciprofloxacin, norfloxacin | inhibits nucleic acid synthesis; for Pseudomonas aeruginosa & other aerobes |
| sulfa drugs (sulfonamides) action & uses sulfamethoxazole | analogue of PABA (intermediate in folate synthesis); for UTI, enteric infections, used w/ trimethoprim (Bactrim, etc.) |
| streptogramins action & uses quinupristin/dalfopristin | inhibits protein synthesis; for GP, especially VRE |
| oxazidinones action & uses linezolid | inhibits protein synthesis; for GP, including those resistant to other abx |
| define: narrow spectrum | only certain groups covered |
| define: broad spectrum | GP & GN coverage |
| define: selective toxicity | action against only microbial structures (70S ribosome, cell wall, etc.) |
| define: bactericidal action | kills bacteria w/out host immune help |
| define: bacteriostatic action | reversible inhibition (ultimate destruction depends on host defenses) |
| synergism | combined better than the sum 1+2=4 |
| antagonism | one decreases activity of another 1+2=1 |
| Kirby-Bauer susceptibility testing | Mueller-Hinton agar, depth 4mm, pH 7.2-7.4, physiologic conc of Ca++ & Mg++, 35 C, ambient air, 10^8 orgs (McFarland 0.5), QC weekly & w/ each new lot of agar or discs (E.coli, S.aureus, P.aeruginosa) |
| broth susceptibility testing: MIC & MBC | MIC (min inhibitory): lowest concentration that prevents in vitro growth, 1st dilution tube w/ no growth; MBC (min bacteriostatic): lowest conc that results in >99.9% killing, subculture tubes near MIC to find 1st plate w/ no growth |
| modified susceptibility testing for slow-growing or fastidious orgs | haemophilus test medium; supplemented MH for S.pneumo; supplemented CG agar base for N.gonorr |
| extended spectrum beta-lactamase (ESBL) definition | resistance enzymes to extended-spectrum (3rd gen) cephalosporins & monobactams (all penicillins, cephalosporins, aztreonam should report as resistant) - don't affect cephamycins; esp consider Escherichia & Klebsiella |
| D-test | for detection of inducible clindamycin resistance; clindamycin 2ug disks & erythromycin 15ug disks; inducible strains form "D"-shaped zone of inhibition |
| how to detect MRSA | zone of <10mm w/ oxacillin 1ug/ml disk on Mueller-Hinton; molecular tests for mecA gene |
| disk diffusion: what causes: tetracycline zone too large, clindamycin too small w/ E.coli or S.aureus controls | pH of agar too low |
| disk diffusion: what causes: tetracycline zone too small, clindamycin too large w/ E.coli or Saureus controls | pH of agar too high |
| disk diffusion: what causes: aminoglycoside zone too small w/ P.aeruginosa control | Ca++ and/or Mg++ too high in agar |
| disk diffusion: what causes: aminoglycoside zone too large w/ P.aeruginosa control | Ca++ and/or Mg++ too low in agar |
| disk diffusion: what causes: zones universally too large on control plates | - inoculum too light - nutritionally poor medium - slow-growing org (not seen w/ controls) - agar depth too thin |
| disk diffusion: what causes: zones universally too small on control plates | - inoculum too heavy - agar depth too thick |
| disk diffusion: what causes: methicillin zone decreasing over days/weeks w/ control orgs | methicillin degrading during refrigerator storage |
| disk diffusion: what causes: methicillin zone indeterminate in disk test | methicillin being degraded by strong beta-lactamase producing Staphylococci |
| disk diffusion: what causes: colonies w/in zone of inhibition | mixed culture; resistant mutants w/in zone |
| disk diffusion: what causes: "zone w/in a zone" phenomenon | - swarming Proteus - feather edges of zones around penicillin or ampicillin disks usually occur w/ beta-lactamase neg strains of S.aureus - beta-lactamase pos H.influenzae w/ penicillin or ampicillin |
| Blood Agar (BA, BAP) purpose | most bacteria; determines hemolytic reactions |
| chocolate agar purpose | Haemophilus & Neisseria sp; enriched w/ hemoglobin or IsoVitalleX |
| phenylethyl alcohol agar (PEA) purpose | selects for GPC & anaerobic GNR |
| Columbia Collistin-Nalidixic Acid (CNA) agar purpose | selects for GPC |
| Thayer-Martin Agar purpose | N.gonorrhoeae & N.meningitidis |
| CAMPY-blood agar purpose | Campylobacter sp |
| thioglycolate broth purpose | "back up" for anaerobes |
| lowenstein-jensen agar purpose | Mycobacterium sp |
| middlebrook 7H10 agar purpose | Mycobacterium sp |
| petragnani agar purpose | Mycobacterium sp |
| bacterioides bile esculin agar (BBE) purpose | selects for B.fragilis group (black colonies) |
| kanamycin-vancomycin laked blood (KVLB) media purpose | Bacterioides sp (enhances pigment production) |
| cyclosterine-cefoxitin fructose agar (CCFA) prupose | C.difficile |
| cooked or chopped meat medium purpose | anaerobes |
| cold enrichment for which orgs? | Listeria monocytogenes & Yersinia |
| Bordet-Gengou agar purpose | B.pertussis |
| Buffered Charcoal Yeast Exctract (BCYE) medium purpose | Legionella sp |
| Fletcher's Medium purpose | leptospira |
| skirrow agar purpose | Helicobacter pylori |
| thiosulface citrate-bile salts (TCBS) medium purpose | Vibrio sp |
| vaginalis agar (V-agar) (human blood) purpose | Gardnerella vaginalis |
| cystine-tellurite blood (Tinsdale) agar purpose | C.diphtheriae (black colonies) |
| loeffler's medium purpose | C..diphtheriae (enhances grouping & metachromatic granules) |
| Carbohydrate fermentation w/ phenol red: uninoculated/neg; pos | red; yellow |
| esculin: uninoculated/neg; pos | nondescript; black |
| hippurate hydrolysis: uninoculated/neg; pos | no change; purple |
| motility: uninoculated/neg; pos | growth along stab line; blurred stab line |
| 6.5% NaCl broth: uninoculated/neg; pos | clear; turbid |
| phenylalanine or tryptophan deaminase: uninoculated/neg; pos | nondescript; green surface after 10% FeCl3 |
| urea: uninoculated/neg; pos | nondescript; bright pink |
| general specimen collection/handling, 7 | 1 material from infection site; 2 optimal time; 3 sufficient quantity; 4 appropriate collection devices; 5 never refrigerate spinal fluids, anaerobic, or GC specimens; 6 prior to abx therapy; 7 set up w/in 2 hrs of collection |
| criteria for specimen rejection, 4 | 1 preservatives used; 2 QNS; 3 dry swab; 4 leaky containers (contaminated specimen, biohazard) |
| blood culture collection, 6 | 1 prep skin w/ 70% -OH & iodine, 2 best time just before fever spike, 3 @ least 2 cx, <4/24hrs, 4 may use abx removal devide (ARD) or resin bottles if pt on abx, 5 Isolator best for fungi and AFBs, 6 must have 1:10 dilution, >10mL ideal for adults |
| light microscopy resolving power & lens magnifications | resolving power 0.2 um; ocular lens 10X; oil immersion lens 100X |
| darkfield purpose | spirochetes; reflected light |
| fluorescence - near UV range purposes | auramine rhodamine, acridine orange, calo-fluor white stains |
| electron microscopy resolution & use | resolution 0.001 um; useful in viral ID |
| culture temps, most plates/Campylobacter/Yersinia | 35-37 C / 42 C / 25-30 C |
| CO2 & humidity requirements for culture | 5-10% CO2, 50-70% humidity |
| anaerobic conditions for pre-reduced media | gas pak jars or anaerobic chamber; 10% H2, 5% CO2, 85% N2, palladium crystals |
| non-selective media | supports most orgs; BAP, CHOC, trypticase |
| selective agar | contains chemicals, dyes, abx to inhibit certain orgs (EMB, MAC, CNA, Campy-blood); may also be differential (HE, SS, XLD, EMB, MAC) |
| Gram Stain components; pos/neg | crystal violet, alcohol/acetone, safranin (iodine mordant, methanol/heat fix); purple/pink |
| Kinyoun & Ziel-Nielson components; pos/neg; use | carbol fuchsin, acid alcohol, methylene blue; pink/blue; acid fast for Mycobacteria |
| Auramine-Rhodamine components; pos/neg; use | auramine and rhodamine (fluorescent stain), acid alcohol, potassium permanganate; orange fluoresc./no fluoresc.; Mycobacteria |
| calcofluor white components; pos/neg; uses | calcofluor white + 10% KOH (to break down debris & mucous); bluish-white fluoresc/no fluoresc; yeast/fungi |
| staphylococcus | "grape-like" clusters, GPC |
| S.aureus 2 key things | coagulase pos; most common pathogen of genus |
| S.aureus common infections 4 | furuncles (boils) & carbuncles; bullous empetigo (blisters); paronychia (nails); post surg wounds & bacteremia |
| S.aureus intoxications 3 | scalded skin syndrome (exfoliation - neonates); toxic shock syndrome (TSST-1) (women ages 12-52); food poisoning (enterotoxin) (sx 1-5 hrs after ingestion of potato salad, cream dishes) |
| S.aureus exotoxins | hemolysins, leukocidins, coagulase & hyaluronidase (spreading factor), nuclease, protease, lipase |
| S.aureus Resistance/sensitivity | - most resistant to penicillin due to plasmid mediated B-lactamase - some sensitive to penicillinase-resistant penicillins (PRP's) (methicillin, oxacillin, etc) - if methicillin-resist S.aureus (MRSA), vancomycin is drug of choice |
| S.aureus Lab Diagnosis | - BAP: soft, opaque, regular colonies 2-3 mm in diam; some B-hemolytic, some pale golden color - growth in 7.5% NaCl & ferment mannitol - catalase & coagulase pos - phage typing & susceptibility profile for epidemiologic studies |
| Coag-neg Staph (S.epi, S.saprophyticus) key things | opportunist in immunocompromised hosts & pts w/ prosthetic valves & devices |
| Catalase Test Reagent, process, key results | 1. reag: 3% H2O2 2. add 1 drop to colony on slide 3. if catalase present, H2O2 broken down to H2O & O2 (bubbles off) 4. use to differentiate Staph (+) from Strep (-) 5. QC each day of use |
| Coagulase Test Reagent, process, key results | 1. reag: EDTA rabbit plasma 2. bound coag: clump on slide (plasma & colony) 3. free coag: gels in tube test (0.5 mL plasma & colony; 35-27 C; 4-12 hrs) 4. agglutination: detect coag & protein A 5. use to differentiate S.aureus (+) from other staph (-) |
| S.aureus: cat/coag; infections; intoxications; notes | cat+, coag+; carbuncles, furuncles, paronychia, wounds, bacteremia; scalded skin syndrome, toxic shock syndrome, gastritis; most B-lactamase+, many MRSA, CA=comm acquired, HA=hospital acquired |
| S.epidermidis: cat/coag; infections; notes | cat+, coag=; endocarditis, prosthetic device infections; most methicillin resistant, sensitive to novobiocin |
| S.saprophyticus: cat/coag; infections; notes | Cat+, coag=; UTI in young women; resistant to novobiocin |
| All Staph is Catalase what? Coag Neg Staph causes what? | All Staph is Catalase +. Coag = Staph causes opportunistic infections in immunocompromised & those w/ prosthetic devices. |
| Streptococcus shape; grouping based on what? | spherical/oval, chains/pairs; Lancefield grouping based on C carbohydrate |
| S.pyogenes (GroupA): hemolysis; streptolysins; toxin; abx note | beta hemolytic, streptolysin S: O2 stable, non-antigenic; Streptolysin O: O2 labile; antigenic. erythrogenic toxin: Scarlet fever rash. Highly sensitive to penicillin |
| S.pyogenes (GroupA): infections & diagnosis | pharyngitis (Strep throat), impetigo, erysipelas, wounds/burns, rheumatic fever (autoimmune sequelae); sensitive to 0.04 units bacitracin disc, typing |
| S.agalactiae (Group B): hemolysis; what it causes; lab diagnosis | narrow zone of B-hemolysis; neonatal sepsis & meningitis, UTI, vaginal infections; serotyping, CAMP rxn (w/S.aureus), Na hippurate positive |
| Strep Group D: there are 2 | S.bovis: BEM +, 0 growth in 6.5%NaCl; Enterococcus: BEM +, growth in 6.5%NaCl |
| S.pneumoniae: morphology, abx, causes:, dx | A-hemolytic crater colonies, mucoid water drop colonies, lancetshaped diplococci; check sens to penicillin w/ OX disc; lobar pneumonia, meningitis, bacteremia, otitis media, conjunctivitis; colony morph, quellung rxn, sens to optochin, bile soluble |
| other alpha Streptococus (viridans group) causes | subacute bacterial endocarditis |
| Neisseria & Moraxella key characteristics 2 | GN diplococci (kidney bean shape), oxidase + |
| N.gonorrhoeae growth req, fermentations, causes, ID | grows on CHOC & Thayer-Martin; 5-10% CO2, may take 48 hrs for growth; ferments glucose; causes gonorrhea; molecular probe |
| N.meningitidis growth req, ferment, transmission, causes | grows on BAP, CHOC, Thayer-Martin, 5-10% CO2; ferm glucose & maltose; transm by respiratory droplets, close contact; meningitis (kids under 3), Waterhouse-Friderichsen syndrome=meningiococcemia |
| bacterial meningitis 3 key lab values | incr neutrophils, decr glucose, incr protein |
| Moraxella (Branhamella) catarrhalis causes, growth, morph, abx | respiratory infections; CHOC & BAP, no MAC; hockey puck colony; asaccharolytic; usually B-lactamase + |
| Acinetobacter spp causes, abx, ID | emerging pathogen; respiratory infections, UTI, or colonizer; resist to many drugs - AST required; growth on MAC, some hemolytic, oxidase= |
| Differentiating Neisseria based on sugar fermentation. Glucose, Maltose, Lactose. | All ferment glucose. Gonorrhoeae = only Glucose. Meningitidis = glucose & Maltose. Lactamica = glucose, maltose, & Lactose. |
| GPR: Corynebacterium diphtheriae key characteristics 5 | small pleomorphic rods w/ clubbed ends. palisade arrangement. metachromatic granules stain red/purple w/ methylene blue. Tinsdale agar: tellurite hydrolysis->black colonies. elek test determines toxin production in vitro. |
| GPR: Corynebacterium diphtheriae Loeffler's media, exotoxin, pseudomembrane, causes: | Loeffler's: enhances metachromatic granules. exotoxin only by lysogenic orgs carrying B phage. produces pseudomembrane on tonsils, uvula, soft palate. causes diphtheria. |
| GPR: Listeria monocytogenes | sm colonies, narrow B-hemolysis, cat+, tumbling motility, umbrella motility @ RT, not37C, bile esculin+. causes neonatal meningitis & sepsis, sepsis in immunocompromised. |
| GPR: Erysipelothrix | non-motile, cat=, "test tube brush" growth in gelatin, H2S+ in TSI; occupational infection for fishermen, butchers, veterinarias, rose growers |
| GPR: Bacillus sp (sporeformers) key characteristics | large, ground glass colonies; some beta hemolysis; catalase+; large GP to variable in chains w/ spores |
| GPR: Bacillus anthracis morphology, motility, hemolysis, disease | very long chains bamboo shoots; medusa head colonies; non-motile; non-hemolytic; anthrax - cutaneous, pulmonary, GI |
| GPR: Bacillus cereus disease | food poisoning due to preformed toxin - "fried rice" stool |
| GNR: Enterobacteriaceae general characteristics | peritrichous flagella; ferment GLU; reduce NO3-NO2; most oxidase=; Ag: flag=H, envelope=K, cell wall=LPS=O; all possess LPS endotoxin, some produce exotoxins |
| GNR Enterobact E.coli key characteristics, diseases | indole & lactose+; IMViC = ++--; most common cause of UTI in females; intestinal infections (another card); K1 strains can cause neonatal meningitis |
| GNR Enterobact E.coli Intestinal Infections | enterotoxigenic ETEC-LT toxin (heat labile), ST toxin (heat stable) EIEC; enteroinvasive: penetrate epi cells in bowel, may be lac=; enterohemorrhagic EHEC-O157:H7: shigella-like toxin, food poisoning undercooked meat, hemolytic uremic syndrome (HUS) |
| GNR Enterobact Shigella key characteristics & diseases | lactose=, non-motile, anaerogenic, <200 orgs needed for disease; Bacillary dysentery - penetrate epi cells in sm intestine |
| Remember Shigella Groups A - D | S.dysenteriae = Group A (1st alphabetically) S.flexneri = Broup B (not "B"oydii) S.boydii = Group C ("boyd ee ee" = "C") S.sonnei = Group D (last alphabetically) |
Created by:
jwesoloski14
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