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m(ASCP)
Polansky
| Term | Definition |
|---|---|
| Biosafety Level I (BSL-I) | - min risk, 0 disease - Bacillis subtilis, Mycobacterium gordonae, soil microbes - standard precautions |
| Biosafety Level II (BSL-II) | - mod risk, common pathogens - E. coli, Salmonella, HIV, HBV, flu - BSC I or II, PPE, autoclave available, limited access, most micro labs |
| Biosafety Level III (BSL-III) | - high risk, serious/lethal disease via inhalation, tx available - B.anthracis, Francisella, Brucella, Mycobacterium tuberculosis, Rickettsia rickettsii, Coxiella burnetii, mold stages of systemic fungi - BSL-II plus neg. air flow & sealed windows |
| Biosafety Level IV (BSL-IV) | - extreme risk, life-threatening disease, transmitted by aerosol, no vacc or tx - Ebola virus, Lassa virus, other hemorrhagic fevers - class III bsc, full-body air-supplied pos pressure suit, indep unit w/ ventilation and waste management |
| Category A | CDC Classification of Biological Agents - highest priority - easily disseminated - high mortality, potential for major impact - B.anthracis, Y.pestis, F.tularensis, C.botulinum toxin, smallpox, hemorrhagic fever viruses |
| Category B | CDC Classification - 2nd priority mod to disseminate - mod illness, low mortality - Brucella, Salmonella, Shigella, E. coli O157:H7, Burkholderia, C.psittaci, C.burnetii, C.perfringens tox, ricin tox, staph enterotixin B, R.prowazekii, viral enceph |
| Category C | CDC Classification of Biological Agents - 3rd highest - could be engineered for mass dissemination - high morbidity/mortality, major impact - Nipah virus, hantavirus |
| BSC Class I | - open front, unsterilized room air enters, HEPA filter - minimal personnel protection, no work surface protection |
| BSC Class II | - laminar flow, variable sash opening, air passes through 1 HEPA filter before work surface, 2nd HEPA filter later - most commonly used in hospital micro labs, protection for worker and work |
| BSC Class III | - completely enclosed, negative pressure, air is filter sterilized in and out, gloves attached to front - max protection, labs that work w/ extremely hazardous orgs |
| Steam Under Pressure | Sterilization & Disinfection Methods - most practical & dependable - 15 lbs of pressure, 15 min - Autoclave - 121C - kills spores |
| Boiling | Sterilization & Disinfection Methods - not reliable - spores may not be killed |
| Pasteurization | Sterilization & Disinfection Methods - used in food industry to kill foodborne pathogens - doesn't sterilize - liquid heated to 71.7C, 15 sec |
| Hot Air Sterilization | Sterilization & Disinfection Methods - used when steam may damage or fail to penetrate - 2 hrs, 170C - kills spores |
| Incineration | Sterilization & Disinfection Methods - sterilize inoculating loops & biomedical wastes |
| Filtration | Sterilization & Disinfection Methods - used to sterilize thermolabile liquids (e.g., urea broth) - HEPA filters in BSC |
| Radiation | Sterilization & Disinfection Methods - UV light, used in BSC |
| Alcohols | Sterilization & Disinfection Methods - ethanol & isopropanol (70-80%) frequently used as antiseptics & disinfectants - doesn't kill spores |
| Chlorine | Sterilization & Disinfection Methods - Sodium Hypochlorite (household bleach) among most effective against HIV & HBV - 10% sln, prepare daily |
| Formalin | Sterilization & Disinfection Methods - aqueous sln of formaldehyde - 5-10% formalin used to preserve & fix specimens - exposure must be monitored |
| Glutaraldehyde | Sterilization & Disinfection Methods - effective against most vegetative cells, not mycobacteria - cold sterilization of items damaged by heat, e.g., inhalation therapy equipment, equipment w/optical lenses such as endoscopes |
| Hydrogen Peroxide | Sterilization & Disinfection Methods - 3% sln used as antiseptic |
| Iodine & Iodophors | Sterilization & Disinfection Methods - Iodophors (e.g., povidone-iodine) release iodine slowly, less irritating, non-staining - effective skin antiseptics |
| Phenolics | Sterilization & Disinfection Methods - don't kill spores - other disinfectants evaluated by comparing effectiveness "phenol coefficient" |
| Quaternary ammonium compounds | Sterilization & Disinfection Methods - effective against wide variety of vegetative bacteria - not effective against spores, mycobacteria, or nonenveloped viruses - used on floors, walls, furniture |
| Ethylene Oxide | Sterilization & Disinfection Methods - widely used in hospitals on materials that can't withstand steam |
| Bacterial Exotoxin | Bacterial Toxins - BY: Gram positives - WHERE: w/in & w/out - COMP: protein - EFFECT: systemic - TOXICITY: high - STABILITY TO HEAT: unstable - ANTIGENICITY: high - STIM ANTITOXIN PROD: yes - CONVERT TO TOXOID: yes |
| Bacterial Endotoxin | Bacterial Toxins - BY: Gram negatives - WHERE: w/in - COMP: LPS - EFFECT: local - TOXICITY: low - STABILITY TO HEAT: stable - ANTIGENICITY: low - STIM ANTITOXIN PROD: no - CONVERT TO TOXOID: no |
| Specimen Collection Guidelines | - acute phase, before abx - aspir/tiss>swabs - swabs: Dacron or poly & plastic - ANER: aspirates>swabs - avoid normal flora - not QNS - sterile leakproof - don't label lid - 2ary container - req no contam - remove needles - >30m: preserve |
| Anaerobes | Specimen Preservation & Storage - anaerobic transport systems, no O2 |
| CSF | Specimen Preservation & Storage - 25 or 35C, no fridge |
| Neisseria gonorrhoeae | Specimen Preservation & Storage - Amies transport medium w/ charcoal, transgrow, JEMBEC plates, Gono-pak, BioBag (best to inoculate medium directly) - 25 deg C, no fridge |
| Sputum | Specimen Preservation & Storage - 4C |
| Stool for Culture | Specimen Preservation & Storage - transport media, e.g., Cary-Blair w/in 2 hours - 4 deg C, no delay, fridge may kill Shigella |
| Stool for OAP | Specimen Preservation & Storage - polyvinyl alcohol (PVA), 10% formalin, sodium acetate-acetic acid-formalin (SAF), merthiolate-iodine-formalin (MIF), others w/out formalin or mercury - formed specimens @ 4C, soft or liquid: examine or preserve |
| Urine | Specimen Preservation & Storage - transport tubes w/ boric acid-glycerol if not processed w/in 2 hrs of collection - 4 deg C, <24 hrs |
| Viruses | Specimen Preservation & Storage - VTM - 4 deg C; if >24 hrs, -70 deg C |
| Fragile Orgs | - anaerobes - Chlamydia - Haemophilus influenzae - Neisseria gonorrheae - Neisseria meningitidis - Salmonella - Shigella - Streptococcus pneumoniae - viruses - parasites |
| Rejection Criteria | - unlabeled or improperly labeled - improper site - >2hrs w/out preservation - temp during transport or storage - leaking - nonsterile containers - improper transport medium - swab dried out - wrong swab - needle attached - inappropriate source |
| Crystal Violet | Gram Stain Reagents - basic dye - stains all bacteria purple |
| Iodine | Gram Stain Reagents - mordant - fixes crystal violet to cells |
| 95% ethanol, acetone, or combo | Gram Stain Reagents - decolorizer - removes crystal violet from GN |
| Safranin | Gram Stain Reagents - counterstain - stains GN pink |
| Gram Positive | Staining Properties - thick peptidoglycan w/ teichoic acid & lipoteichoic acid (cross-links prevent decolorization) - stained purple by crystal violet |
| Gram Negative | Staining Properties - thin peptidoglycan w/ proteins, phospholipids, & LPS; decolorizer incr permeability of lipid-rich cell wall - crystal violet washes out - stains pink by safranin |
| Supportive | Types of Media - Supports growth of most non-fastidious bacteria - nutrient agar, trypticase soy agar |
| Enrichment | Types of Media - contains added growth factors, e.g., blood, vitamins, yeast extract - SBA, CHOC, BHI, buffered charcoal-yeast extract agar |
| Selective | Types of Media - contains dyes, bile salts, alcohols, acids, or abx to inhibit growth - Columbia colistin-nalidixic acid (CNA) agar, eosin methylene blue (EMB), MacConkey, Hektoen enteric (HE) , xylose lysine deoxychocolate (XLD), Thayer-Martin |
| Differential | Types of Media - provides distinct colonial appearances based on biochem rxns (e.g., lactose fermentation, H_2_S production) - EMB, MacConkey, HE, XLD |
| Sheep Blood Agar (SBA) | Routine Media - E, D - most nonfastidious bacteria - tryptic soy agar w/ 5% sheep blood; differentiation of hemolysis |
| Chocolate Agar (CHOC) | Routine Media - E - Haemophilus & Neisseria - supplies X & V factors; incubate in high CO_2 |
| Columbia Colistin-Nalidixic Acid Agar (CNA) | Routine Media - S - Gram Pos - Colistin & Nalidixic acid supress most Gram Neg; contain 5% sheep blood |
| Phenylethyl Alcohol Agar (PEA) | Routine Media - S - GPC & anaerobic GNR - phenylethyl alcohol inhibits enteric GNR; contains 5% sheep blood |
| Group A-Selective Strep Agar w/ 5% sheep blood (SSA) | Routine Media - S - Group A strep from resp sources - contains abx to suppress normal throat flora. GBS will also grow. |
| Eosin Methylene Blue (EMB) | Routine Media - S, D - Enteric GNR - eosin & methylene blue inhibit GP; LF green-black or purple; E.coli produces green metallic sheen; NLF colorless |
| MacConkey Agar (MAC) | Routine Media - S, D - Enteric GNR - bile salts & crystal violet inhibit most GP; LF pink; NLF colorless |
| Sorbitol MacConkey Agar (SMAC) | Routine Media - S - E. coli O157:H7 - [above] doesn't ferment sorbitol; colorless colonies; some labs stopped using because non-O157 serotypes can be pathogens |
| Hektoen enteric (HE) Agar | Routine Media - S, D - for Salmonella & Shigella in stool - bile salts, bromothymol blue, & acid fucsin inhibit normal GI flora - nonpath orange to salmon pink - NLF green to blue-green - H_2_S pos colonies have black precipitate |
| Salmonella-Shigella (SS) Agar | Routine Media - S - Salmonella & Shigella in stool - brilliant green & bile salts inhibit other enterics - Salmonella & Shigella NLF (colorless) - Salmonella H_2_S pos (black center) |
| Gram-negative broth (GN) | Routine Media - E, S - Salmonella & Shigella from stool & rectal swabs - deoxychocolate & citrate salts retard growth of GP - sub onto S, D agar after 6-8 hr & 18-24 hrs incubation - use of enrichment broths for stool cx is decreasing |
| Deocychocolate-citrate Agar | Routine Media - S - Salmonella & Shigella - other enterics inhibited |
| Campylobacter broth | Routine Media - E - Campylobacter from stool - sub to Campy-selective agar after overnight incubation @ 4degC |
| Campylobacter Blood Agar (Campy BAP) | Routine Media - E, S - Campylobacter from stool - incubate in high CO_2 @ 42degC |
| Modified Thayer-Martin (TM) | Selective media for Neisseria gonorrhoeae & Neisseria meningitidis from sites w/ normal flora - vancomycin, colistin, nystatin & trimethoprim inhibit other bact & fung - incubate in high CO_2 - some N.gonorrhoeae may be inhibited |
| Martin-Lewis | Selective Media for Neisseria gonorrhoeae & Neisseria meningitidis from sites w/ normal flora - similar to TM, but different abx - inhibits yeast better - incubate high CO_2 |
| New York City medium (NYC) | Selective Media for Neisseria gonorrhoeae & Neisseria meningitidis from sites w/ normal flora - incubate high CO_2 - some N.gonorr inhibited by abx - genital mycoplasmas will grow |
| GC-LECT | Selective Media for Neisseria gonorrhoeae & Neisseria meningitidis from sites w/ normal flora - abx to inhibit GN & GP bacteria & yeast |
| JEMBEC plates | Selective Media for Neisseria gonorrhoeae & Neisseria meningitidis from sites w/ normal flora - for transport & growth of N.gonorr - contain Neisseria-selective medium - resealable polyethylene bag & CO2 gas tablet - no need to transfer to cx plate |
| Cysteine-tellurite Blood Agar | Special Bacteriologic Media - D --> Corynebacerium diphtheriae - C.diphtheriae produce black colonies |
| Loeffler Medium | Special Bacteriologic Media - E --> C. diphtheriae - promotes metachromatic granules |
| Tindale Agar | Special Bacteriologic Media - S, D --> C.diphtheriae - Corynebacterium spp produce grey-black colonies due to reduction of tellurite, surrounded by brown halo |
| Bismuth Sulfite Agar | Special Bacteriologic Media - S --> Salmonella - Bismuth sulfite & brilliant green inhibit most others - S.typhi = black w/ metallic sheen - others light green - some salmonellae may be inhibited |
| Cefsulodin-irgasan-novobiocin (CIN) Agar | Special Bact... Media - S --> Yersenia enterocolitica, Aeromonas, & Plesiomones shigelloides - crystal violet inhibits GN - Novobiocin inhibits GPC - Cefsulodin inhibits GP & GN - Y.enterocolitica ferments mannitol, red "bull's-eye" w/ colorless halo |
| Alkaline Peptone Water (APW) | Special Bacteriologic Media - E --> vibrio from stool - alkaline pH suppresses commensals - subbed to TCBS |
| Thiosulfate Citrate Bile Salts Sucrose (TCBS) Agar | Special Bacteriologic Media - S --> Vibrio - high pH inhibits most bact - V.cholerae ferments sucrose = yellow colonies - V.parahaemolyticus & V.vulnificus don't ferment sucrose = blue-green colonies |
| Bordet-Gengou Agar | Special Bacteriologic Media - S, E --> Bordetella pertussis - potato-glycerol-based medium enriched w/ blood - contaminants inhibited by methicillin - "cough plate" - Bordetella colonies resemble mercury droplets |
| Regan-Lowe Agar | Special Bacteriologic Media - S --> B.pertussis - charcoal agar supp w/ horse blood, cephalexin & amphotericin B |
| Buffered charcoal-yeast extract (BCYE) Agar | Special Bacteriologic Media - E --> Legionella - yeast extract & L-cysteine enhance growth of Legionella - charcoal absorbs toxic compounds |
| Human Blood Bilayer Tween (HBT) Agar | Special Bacteriologic Media - S, D --> G.vaginalis - incubate high CO_2 48 hrs - beta hemolytic |
| Aerobe | Aerotolerance Test - growth in SBA (RA) & CHOC (CO2) - no growth in SBA (ANER) |
| Capnophilic Aerobe | Aerotolerance Test - growth in CHOC (CO_2) - no growth in SBA (aer) or (aner) |
| facultative anaerobe | Aerotolerance Test - growth in SBA (aer) and (aner), and CHOC (CO_2) |
| obligate anaerobe | Aerotolerance Test - growth on SBA (aner) - no growth on SBA (aer) or CHOC (CO_2) |
| aerotolerant anaerobe | Aerotolerance Test - growth on SBA (aner) and CHOC (CO_2) - no growth on SBA (aer) |
| Orgs Req Incubation in Incr CO_2 | - Campylobacter - Haemophilus - Helicobacter - Moraxella catarrhalis - Mycobacterium - Pathogenic Neisseria |
| Alpha | Hemolytic Rxns on SBA - green zone around colony, narrow or wide - partial lysis - Streptococcus pneumoniae, viridans streptococci, some enterococci |
| Beta | Hemolytic Rxns on SBA - Clear zone around colony, narrow or wide - complete lysis - GAS, GBS, Listeria monocytogenes |
| Gamma (nonhemolytic) | Hemolytic Rxns on SBA - no hemolysis - some enterococci |
| Staphylococcus spp. (1) | Staphylococci (1) - GS: GPC, clusters - on SBA: 1-3mm, round, smooth, convex, glistening, opaque, entire edge, butyrous - catalase+, fermentative, microdase neg, bacitracin res, furazolidone & lysostaphin susc |
| Staphylococcus spp. (2) | Staphylococci (2) - normal: skin, mouth, pharynx, vagina, urethra, GI - facultative anaerobe - most non-S media - salt tolerant |
| Staph aureus (1) | Staphylococci (1) - PATH: suppurative cutaneous infections, TSS, food poisoning - GS: GPC, clusters - on SBA: most beta hemolytic (small), may be golden - coagulase+, ferm. mannitol (yellow on mann. salt agar), usually DNase & thermonuclease + |
| Staph aureus (2) | Staphylococci (2) - 10-60% are carriers - direct contact - 85-90% penicillin resist |
| Coagulase-neg Staphylococci (CNS) (1) | Staphylococci (1) - PATH: opportunistic - common cause of hospital-acquired UTI - GS: GPC, clusters - on SBA: white, usually non-hemolytic - coagulase neg - grows on mannitol salt agar, doesn't ferm - novobiocin sens |
| Coagulase-neg staphylococci (CNS) (2) | Staphylococci (2) - normal: skin & mucous membranes - often contaminant - usually only speciated if from normally sterile site - 50-80% are S.epidermidis |
| Staph saprophyticus (1) | Staphylococci (1) - PATH: UTI in young sexually active females, urethritis & prostatitis in males - GS: GPC, clusters - on SBA: white-slightly yellow, non-hemolytic |
| Staph saprophyticus (2) | Staphylococci (2) - coagulase neg, novobiocin resist, may ferment mannitol - novobiocin only performed when isolated from urine of female |
| Micrococcus (1) | Staphylococci (1) - usually nonpathogenic, found in environ & on skin, mucous membranes, oropharynx - GS: Lg GPC in pairs, tetrads (predominant), clusters - on SBA: often pigmented (bright yellow, orange, pink, tan), high-domed colonies |
| Micrococcus (2) | Staphylococci (2) - catalase+, coagulase neg, oxidative, microdase+ - bacitracin susc, furazolidone & lysostaphin resist - not commonly isolated - must diff from staph - usually only grow aerobically |
| Catalase | Tests for ID of Staphylococci - immediate bubbling - 3% H2O2 -> O2 + H2O - Staph pos; Strep & entero neg - bubbles after 20-30 sec not pos rxn; RBCs in blood agar may produce weak bubbles |
| Coagulase | Tests for ID of Staphylococci - coagulation (tube) or agglutination (slide) in plasma - S. aureus pos - slide = screening = bound coagulase; if neg, tube = free coagulase - largely replaced by latex agglutination |
| Slide agglutination for Staph aureus | Tests for ID of Staphylococci - agglutination of latex beads coated w/ fibrinogen & Abs to protein A (in cell wall of S. aureus) - S. aureus pos - most labs report pos orgs as S. aureus |
| Mannitol salt agar (MSA) | Tests for ID of Staphylococci - ferm of mannitol results in color change pink --> yellow - S. aureus pos - 7.5% salt inhibits most orgs other than staph; all staph grow on MSA; rarely used or ID of S. aureus because other species ferm mannitol |
| Novobiocin susceptibility | Tests for ID of Staphylococci - orgs resist to novobiocin grow to edge of disk - S. saprophyticus is resist; other CNS susc - performed on CNS isolated from urine |
| Streptococcus spp | Streptococci/Enterococci - GS: oval GPC, chains & pairs - on SBA: <1mm, white to gray, translucent or semiopaque, variable hemolysis - catalase neg, facultative anaerobes, req E media, chaining best in broth cultures |
| Group A Strep (GAS) (1) | Streptococci/Enterococci (1) - PATH: 90% of strep infections, strep sore throat, rheumatic fever, glomerulonephritis, scarlet fever (scarlatina), erysipelas, puerperal sepsis, impetigo - GS: oval GPC, chains |
| Group A Strep (GAS) (2) | Streptococci/Enterococci (2) - on SBA: pinpoint, gray-white, translucent, usually beta-hemolytic |
| Group A Strep (GAS) (3) | Streptococci/Enterococci (3) - bacitracin sens, SXT resist, PYR+, most common S.pyogenes, hemolysis: O2-stable streptolysinS & -labile stretolysinO, to detect spp w/ O only, stab agar, coverslip on inoculum or incubate ANER |
| Group B Streptococci (GBS) (1) | Streptococci/Enterococci (1) - PATH: most common cause of neonatal septicemia & meningitis - GS: oval GPC, chains - on SBA: slightly larger than GAS, gray/white, narrow diffuse beta hemolysis, maybe non- - SXT & bacitracin resist, Na hippurate+, CAMP+ |
| Group B Streptococci (GBS) (2) | Streptococci/Enterococci (2) - S.agalactiae: vag & rectal swabs, preg F 35-7wk, in selective broth (LIM, GBS broth, StrepB Carrot Broth) |
| Group D streptococci, nonenterococci | Streptococci/Enterococci - PATH: normal in GI tract, nosocomial UTI, wound infec, bacteremia - GS: oval GPC, chains - on SBA: usually non-hem, maybe alpha hem - hydrolyzes esculin - most common: S.gallolyticus (formerly S.bovis) |
| Enterococcus (1) | Streptococci/Enterococci (1) - PATH: normal in mouth GI female genital, nosocomial UTI wound infec bacteremia - GS: oval GPC, pairs & chains - on SBA: usually alpha or nonhem, rarely beta hem - hydrolyzes esculin, grows in 6.5%NaCl broth, PYR+ |
| Enterococcus (2) | Streptococci/Enterococci (2) - Lancefield group D, 80% are E.faecalis, 15%E.faecium |
| Streptococcus pneumoniae (1) | Streptococci/Enterococci (1) - PATH: normal URT, most common cause of comm-acquired pneumo, otitis media & meningitis in adults, infects sinuses & eyes - GS: football- or lancet-shaped GPC, usually pairs, maybe single/short chain, freq encapsulated |
| Streptococcus pneumoniae (2) | Streptococci/Enterococci (2) - on SBA: round, translucent, glistening, dome shaped when young, central depression w/ age (umbilicate) due to autolysis, alpha hemo, if encap: mucoid - bile solu+, optochin sens |
| Streptococcus pneumoniae (3) | Streptococci/Enterococci (3) - no Lancefield group, >80 serotypes based on capsular Ag |
| Viridans streptococci | Streptococci/Enterococci - PATH: norm oral resp GI, opportunistic path, freq subacute bact endocarditis, gingivitis & dental caries - GS: GPC chains - on SBA: usually alpha hemo, maybe beta or nonhemo - optochin resist, bile solu neg |
| Microaerophilic Streptococcus spp (formerly S.milleri group) (1) | Streptococci/Enterococci (1) - PATH: normal mouth GI GU, infec assoc w/ trauma/surgery, most common abd head neck - GS: GPC pairs & chains - on SBA: 1/2 size of other strep, usually beta hemo, can be alpha or non hemo, butterscotch or caramel odor |
| Microaerophilic Streptococcus spp (formerly S.milleri group) (2) | Streptococci/Enterococci (2) - usually Lancefield group F, PYR neg, VP pos - 3 spp: S.anginosus, S.constellatus, S.intermedius - in Viridans group, req 10% CO2, grow better ANER |
| Bacitracin disk | Beta Hemolytic Strep ID - zone of inhibition after overnight incubation = susc to bacitracin - GAS susc; GBS resist - aka A disk; on SBA; no longer recommended because some GCS & GGS are susc; replaced by PYR & serogrouping by latex agglutination |
| Trimethoprim-sulfamethoxazole (SXT) disk | Beta Hemolytic Strep ID - orgs resist to SXT grow up to disk - GAS & GBS resist; GCS & GGS susc - used w/ bacitracin disk to diff GAS from C or G; overnight incubation |
| Pyrrolidonase (PYR) test | Beta Hemolytic Strep ID - if PYR is hydrolyzed, red color after add of color developer - GAS pos (also enterococci) - more specific than bacitracin for GAS; GAS is only beta-hemo strep that's PYR pos; disk test takes minutes |
| CAMP test (1) | Beta Hemolytic Strep ID (1) - GBS makes extracellular protein enhances hemo of beta-hemo S. aureus on SBA - GBS pos |
| CAMP test (2) | Beta Hemolytic Strep ID (2) - classic: streak across Beta-lysin-producing S. aureus or disk; incubate ambient overnight (no CO2) arrow-head hemo @ intersection - Rapid: drop of beta-lysin on SBA colonies; incubate 20min |
| Hippurate hydrolysis | Beta-Hemolytic Strep ID - orgs that produce hippuricase (hippurate hydrolase) hydrolyze sodium hippurate to benzoate & glycine - GBS pos - alt to CAMP - 2-hr test available |
| Slide Agglutination Tests | Beta-Hemolytic Strep ID - latex particles coated w/ group-specific Ab agglutinate in presence of bacterial Ag - Orgs agglutinate in corresponding antisera - Tests commercially available for rapid ID of groups A, B, C, D, F, G strep & S.pneumoniae |
| Optochin disk (P disk) | Alpha-hemolytic Strep ID - zone of inhib >14mm w/ 6-mm disk or >16mm w/ 10-mm disk = susc - S. pneumo susc; viridans strep resist - contains ethylhydrocupreine hydrochloride; on SBA lawn; incubate in CO2; zone <14mm, ID as S.pneumo only if bile sol |
| Bile solubility | Alpha-hemolytic Strep ID - bile salts (e.g., sodium deoxy-chocolate) cause lysis of some orgs - S. pneumoniae pos - can be performed in broth or on colony; pos = clearing of broth or disappearance of colony; results in <30 min |
| Pyrrolidonase (PYR) Test | ID nonhemolytic Strep/Enterococci - if PYR is hydrolyzed, red color after add of color developer - Enterococci & GAS pos |
| Bile-esculin agar | ID nonhemolytic Strep/Enterococci - orgs that grow in 40% bile & produce esculinase hydrolyze esculin, black precipitate - GDS & enterococci pos - bile inhibits GPs other than GDS & enterococci |
| Esculin test | ID nonhemolytic Strep/Enterococci - orgs that produce esculinase hydrolyze esculin on disk, producing dark spot - GDS & enterococci pos - doesn't test for bile tolerance |
| 6.5% NaCl broth | ID nonhemolytic Strep/Enterococci - orgs that can grow in 6.5% NaCl produce turbidity - enterococci pos (also GBS) - GDS neg |
| Staph aureus | Antibiograms of GPC - significant resist to multiple abx, including vancomycin - usually resist to penicillin & ampicillin - MRSA resist to all beta-lactams - routine susc testing |
| GAS | Antibiograms of GPC - universally susc to penicillin, cephalosporins, vancomycin - no routine susc testing |
| GBS | Antibiograms of GPC - susc to penicillin, cephalosporins, vancomycin - no routine susc testing; may be done when isolated from baby |
| Streptococcus pneumoniae | Antibiograms of GPC - increasingly resist to penicillin & others; susc to vancomycin - routine susc testing |
| GDS | Antibiograms of GPC - susc to penicillin, vancomycin - routine susc testing |
| enterococci | Antibiograms of GPC - ususally resist to penicillin & several other common abx; most susc to vancomycin, but VRE becoming more common in hospitals - routine susc testing |
| Bacillus spp (1) | Aerobic non-spore forming GPR (1) - PATH: usually contam - GS: lg w/ square ends, maybe chains, oval, subterminal spores, not swollen, like bamboo, spores maybe not seen in direct smear |
| Bacillus spp (2) | Aerobic non-spore forming GPR (2) - CULT: lg, spreading, beta hemo, irregular edges (Medusa head), whitish/grey, maybe pigmented - catalase+, most are motile, must rule out B.anthracis |
| Bacillus anthracis (1) | Aerobic non-spore forming GPR (1) - among most highly pathogenic microorgs; anthrax; from contam hides, wool, meat; rare in US - large, adherent, nonhem, flat-slightly convex, irregular border, ground-glass, comma project; stands like beaten egg white |
| Bacillus anthracis (2) | Aerobic non-spore forming GPR (2) - nonhemolytic (beta hemolysis rules out); catalase pos; nonmotile; capsules seen in CSF & blood smears - potential bioterrorism org; handle in BSC; submit to public health lab for confirmation |
| Bacillus cereus | Aerobic non-spore forming GPR - often contam; local & systemic infect, food pois - GS & CULT: same as B.anthracis except hemolytic - motile - 2nd most import path in genus |
| Corynebacterium spp | Aerobic Non-Spore-Forming GPR - opportunistic pathogens - irregular, slightly curved, nonparallel sides, chinese letter/picket fence (palisades) - facultative anaerobes; grow on most media - catalase+; nonmotile |
| Corynebacterium diphtheriae | Aerobic Non-Spore-Forming GPR - rare in US - gray-black w/ brown halos on Tindale agar; black on cystine tellurite; Loeffler stim metachromatic granules - catalase+; nonmotile - ID req demo of toxin; mod Elek immunoprecip test; PCR TOXgene |
| Corynebacterium jeikeium | Aerobic Non-Spore-Forming GPR - hospital-acquired pathogen - GS & culture same as other commensal Coryne - catalase+; rapid sucrose urea (RSU) neg - most commonly isolated diphtheroid; highly resist to abx |
| Listeria monocytogenes (1) | Aerobic Non-Spore-Forming GPR (1) - meningitis & septicemia in newborns & immunocompromised; food pois - GS: parallel sides, rounded ends, coccobacillary; singles, chains, or diphtheroid - tiny w/ narrow indistinct beta hemolysis; translucent, gray |
| Listeria monocytogenes (2) | Aerobic Non-Spore-Forming GPR (2) - catalase+; hippurate hydrolysis+; esculin+; CAMP+(hemol shovel, not arrowhead); tumbling motil; umbrella in motility agar at RT - grows 0.5-45C; cold enrich maybe; catalase diff from GBS; motility diff from diphth |
| Nocardia (1) | Aerobic Non-Spore-Forming GPR (1) - immunocompetent: skin; immunocompromised: invasive pulmonary & disseminated infect - fine branching filaments w/ fragment; often beaded - slow-growing; SBA wrinkled, dry, crumbly, chalky white-orange-tan, beta hemol |
| Nocardia (2) | Aerobic Non-Spore-Forming GPR (2) - catalase pos; partially acid-fast; sulfur granules - N.brasiliensis most common to cause skin infect; N.asteroides most common to cause lung infect |
| Neisseria spp | Neisseria & Moraxella - GNDC, coffee-bean shaped - aerobic or facultative; pathogens are capnophilic - catalase+; oxidase+ |
| N.gonorrhoeae (1) | Neisseria & Moraxella (1) - gonorrhea, salpingitis, newborn ophthalmia - intra- & extra-cell GNDC; dx urethral d/c from sx males; cx confirm req for fem - req high CO2; usually no growth SBA; grows on CHOC & Neisseria-select media |
| N.gonorrhoeae (2) | Neisseria & Moraxella (2) - small, grayish white or tan; 5 colony types; may look like mixed cx - carb use: GLU; superoxol+; can ID w/ monoclonal Ab - susc to drying & cold; no fridge; molec methods for genital; cx pref for non-genital & children |
| N. meningitidis | Neisseria & Moraxella - normal URT; meningitis in YA - SBA, CHOC, & Neisseria-selective; - bluish gray or tan; maybe mucoid - GLU & MAL; ONPG neg - no fridge; BSC; serogroup slide agglut; Ag test avail, poor sens, shouldn't replace cx; vacc avail |
| Commensal Neisseria spp | Neisseria & Moraxella - normal URT; rarely cause disease - GNDC - SBA & CHOC@RT, nutrient agar@35C; some on S: no high CO2; maybe pigmented - no speciate resp specimens; must diff from path Neisseria if isolated on selective media or sterile site |
| N.lactamica | Neisseria & Moraxella - normal URT - GNDC - selective media; like N.meningitidis but smaller - glu, maltose & lactose (slow); ONPG+ - commensal Neisseria; easily misID'ed as N.meningitidis; may react w/ meningococcal typing sera; ONPG differentiates |
| Moraxella catarrhalis (1) | Neisseria & Moraxella (1) - normal in URT; causes Resp infections in young, old & compromised; otitis media & sinusitis in children - GNDC |
| Moraxella catarrhalis (2) | Neisseria & Moraxella (2) - SBA & CHOC; some may grow at RT or on S; "hockey puck colonies" (can be pushed over agar w/ loop) - catalase+, oxidase+; neg for all sugars & ONPG; DNase+; butyrate esterase+ - DNase & butyrate esterase diff from Neisseria |
| Enterobacteriaceae | Characteristics of _ - non-spore-forming GNR - facultative anaerobes - on SBA: most are large, dull, gray, nonhemolytic - on MAC: LF = pink, NLF = colorless - ferment glu - oxidase neg - most reduce nitrates to nitrites - most catalase pos |
| Oxidase | ID of Enterobacteriaceae - tetramethyl-para-phenylene-diamine dihydrochloride reacts w/ cytochrome C to produce blue or purple color - + = blue or purple - diff Enterobac from nonferm; false+ from iron-containing wire, use platinum wire/wooden stick |
| Nitrate Reduction | ID of Enterobacteriaceae - if org reduces nitrates to -ites, red develops with sulfanilic acid & N,N-dimethyl-L-naphthylamine - + = red or no color w/zinc dust - if no color add zinc; red color after zinc dust = neg rxn; no color after zinc = + rxn |
| Carbohydrate Fermentation | ID of Enterobacteriaceae - when carb fermented, acidic end products; pH indicator - w/ phenol red, red to yellow - frequently tested: glu, lact, sucr, mannose, sorb, mannitol, xyl, adonitol, cellobiose, dulcitol, trehalose; all Enterobact ferment glu |
| ONPG | ID of Enterobacteriaceae - ONPG is changed to orthonitrophenol by beta-galactosidase - pos = yellow - test for slow lactose fermentation; helpful in diff Citrobacter (pos) from Salmonella (neg) |
| H2S production | ID of Enterobacteriaceae - orgs that produce colorless H2S gas from sulfur-containing compounds; H2S reacts w/ iron salt -> black ferrous sulfide - sulfur-containing: sodium thiosulfate, cystine, methionine; diff Salmonella (+) from Shigella (neg) |
| Triple sugar iron (TSI) agar | ID Enterobacteriaceae - sugar ferm = acid - pink=alkaline (K); yellow=acid (A) - butt=GLU ferm/no ferm; slant=lact/sucr ferm/no ferm - black precip=H2S - glu, lac, sucr, phen red, Na thioSate, Fe salt; cap loose; Klinger's iron agar (KIA) same, no su |
| Indole | ID of Enterobacteriaceae - tryptophanase deaminates tryptophan; indole made; Kovacs's reagent = pink complex; + =pink - spot indole test comm available; need tryptophan; use colonies from SBA or CHOC agar, not MAC; + =blue; E.coli & Proteus vulgaris + |
| Methyl red (MR) | ID of Enterobacteriaceae - GLU ferm -> acid; pH indicator - red = + |
| Voges-Proskauer (VP) | ID of Enterobacteriaceae - GLU met -> acetoin - add alpha naphthol & KOH; red = + - VP+ = MR neg (& vice versa) |
| Citrate | ID of Enterobacteriaceae - if citrate = sole C source, pH up, pH indicator - growth, or green/blue = + - false neg if cap tight; Kleb & Enterobacter + |
| Urease | ID of Enterobacteriaceae - urease + urea = ammonia; pH up -> indicator - yellow/pink = + - Proteus & Morganella produce urease rapidly |
| Phenylalanine deaminase (PD) | ID of Enterobacteriaceae - phenylalanine deaminase + phenylalanine = phenylpyruvic acid + ferric Cl = green = + result - Proteus, Providencia, Morganella are + |
| Decarboxylase reactions | ID of Enterobacteriaceae - if AA decarboxylase (ornithine, lysine, arginine) = pH up; pH indicator - yellow/purple = + - ornithine decarboxylase (ODC) diff Kleb (neg) from Enterobacter (+) |
| Motility | ID of Enterobacteriaceae - motile orgs grow away from stab line in motility medium - move away from stab line, or hazy appear after overnight incubation = + - most Enterobact +, except Kleb & Shigella - diff Kleb (neg) & Enterobact (+) |
| O antigen | Enterobacteriaceae Ag - Somatic Ag - cell wall, LPS, heat stable - serological grouping of Salmonella & Shigella |
| H antigen | Enterobacteriaceae Ag - flagellar Ag - flagella, proteins, heat labile, serotype Salmonella |
| K antigen | Enterobacteriaceae Ag - capsular Ag - capsule, polysaccharide, heat labile, may mask O Ag, removed w/ heat - prevent phagocytosis, incr virulence, Vi Ag is K Ag from S.typhi |
| Escherichia coli (1) | Common Enterobacteriaceae (1) - UTI, septicemia, neonatal sepsis & meningitis, some diarrhea - +: lactose, gas, indole, MR, motility - neg: H2S, VP, citrate, PD, urease - pred GI aerobe, most common cause UTI, green met sheen on EMB |
| Escherichia coli (2) | Common Enterobacteriaceae (2) - Presum ID: lactose+, dry on MAC, oxidase neg, indole+ - O157:H7 no sorbitol ferm; colorless on SMAC |
| Shigella | Common Enterobacteriaceae - dysentery (shigellosis); most communicable diarrhea; 1-arily in crowded/substandard conditions: daycare, jail, prison - +: MR - neg: lactose, gas, H2S, VP, citrate, PD, urease, motility |
| Edwardsiella tardia | Common Enterobacteriaceae - opportun, bacteremia, wound infect - +: gas, H2S, indole, MR, motility - neg: lactose, VP, citrate, PD, urease - chief reservoirs: reptiles, fresh H2O fish; infect often from aquatic environ - + indole diff from Salmonella |
| Salmonella | Common Enterobacteriaceae - typhoid, bacteremia, enterocolitis - +: H2S, MR, motility, LDC - neg: lactose, indole, VP, PD, urease, ONPG - poultry, trans by reptiles, S.typi has Vi Ag, only trace H2S, citrate neg, grouped by O Ag, serotyped by H Ag |
| Citrobacter freundii | Common Enterobacteriaceae - nosocomial infect - +: gas, H2S, MR, citrate, motility, ONPG - VP, PD, LDC - lactose variable, ONPG & LDC diff from Salmonella |
| Klebsiella pneumoniae | Common Enterobacteriaceae - pneumonia, UTI, septicemia - +: lactose, gas, VP, citrate, urease (slow) - neg: H2S, indole, MR, PD, motility, ODC - encaps, usually mucoid, k.oxytoca similar to K.pneumo exc indole +, motility & ODC diff from Enterobacter |
| Enterobacter aerogenes & cloacae | Common Enterobacteriaceae - opportun & nonsocomial infect, UTI, RTI, wound infect - +: lactose, gas, VP, citrate, motility, ODC - neg: H2S, indole, MR, PD - maybe mucoid, same IMViC rxn as Klebsiella |
| Serratia marcenscens | Common Enterobacteriaceae - opp path, pneumonia & septicemia in immunosuppressed - +: VP, citrate, motility - neg: lactose, H2S, indole, PD, urease - some red when incubated @ RT |
| Proteus vulgaris & mirabilis | Common Enterobacteriaceae - UTI, wound infect, septicemia - +: H2S, MR, PD, urease, motility - lactose neg - swarming, burned choc odor, mirabilis most common & indole neg, vulgaris indole +, A/A on TSI bc sucrose ferm |
| Morganella morganii | Common Enterobacteriaceae - mainly nosocomial infect, UTI, wound infect - +: indole, MR, citrate, PD, motility - neg: lactose, H2S, VP - GN coccobacilli, bipolar stain, optimal 25-30C |
| Providencia | Common Enterobacteriaceae - UTI, diarrhea - +: indole, MR, citrate, PD, motility - neg: lactose, H2S, VP - P.rettgeri urease + |
| Yersinia enterocolitica | Common Enterobacteriaceae - diarrhea - +: MR, urease - neg: lactose, H2S, VP, citrate, PD - GN coccobacilli, bipolar stain, 25-30C, motile 25C not 35C, CIN agar, incubate 48hrs, red "bulls eye" colonies w/ colorless halo, Y.pestis causes plague |
| Lactose Negative | Key Rxns for Enterobacteriaceae - Shigella - Edwardiella - Salmonella - Citrobacter (some) - Serratia - Proteus - Morganella - Providencia - Yersinia |
| H2S Positive | Key Rxns for Enterobacteriaceae - Edwardsiella - Salmonella - Citrobacter - Proteus |
| VP Positive | Key Rxns for Enterobacteriaceae - Klebsiella - Enterobacter - Serratia |
| PD Positive | Key Rxns for Enterobacteriaceae - Proteus - Morganella - Providencia - Klebsiella (slow) |
| Urease Positive | - Proteus - Morganella - Providencia rettgeri |
| Nonmotile @ 35C | Key Rxns for Enterobacteriaceae - Shigella - Klebsiella - Yersinia (motile @ 22C) |
| Escherichia coli | Appearance of Enterobacteriaceae on Media - TSI: A/A, gas - MAC: flat, dry pink w/ darker pink halo - HE: yellow - XLD: yellow |
| Shigella | Appearance of Enterobacteriaceae on Media - TSI: K/A - MAC: colorless - HE: green - XLD: colorless |
| Edwardsiella | Appearance of Enterobacteriaceae on Media - TSI: K/A, gas, H2S - MAC: colorless - HE: colorless - XLD: red, yellow, or colorless w/ or w/out black centers |
| Citrobacter | Appearance of Enterobacteriaceae on Media - TSI: A/A or K/A, gas, w/ or w/out H2S - MAC: colorless @ 24hr, may become pink @ 48hr - HE: colorless - XLD: red, yellow, or colorless w/ or w/out black center |
| Salmonella | Appearance of Enterobacteriaceae on Media - TSI: K/A, gas, H2S - MAC: colorless - HE: green - XLD: red w/ black center |
| Klebsiella | Appearance of Enterobacteriaceae on Media - TSI: A/A, gas - MAC: pink, mucoid - HE: yellow - XLD: yellow |
| Enterobacter | Appearance of Enterobacteriaceae on Media - TSI: A/A, gas - MAC: pink, maybe mucoid - HE: yellow - XLD: yellow |
| Serratia | Appearance of Enterobacteriaceae on Media - TSI: K/A - MAC: colorless @ 1st, turning pink; S.marcescens may have red pigment @ RT - HE: colorless - XLD: yellow or colorless |
| Proteus | Appearance of Enterobacteriaceae on Media - TSI: K/A (mirabilis) A/A (vulgaris), gas, H2S - MAC: colorless, may swarm - HE: colorless - XLD: yelow or colorless, w/ or w/out black centers |
| Morganella | Appearance of Enterobacteriaceae on Media - TSI: K/A, gas - MAC: colorless - HE: colorless - XLD: red or colorless |
| Providencia | Appearance of Enterobacteriaceae on Media - TSI: K/A - MAC: colorless - HE: colorless - XLD: yellow or colorless |
| Yersinia | Appearance of Enterobacteriaceae on Media - TSI: yellow/orange - MAC: colorless to peach - HE: salmon - XLD: yellow or colorless |
| Enterohemorrhagic (EHEC) (1) aka Shiga toxin-producing (STEC) or verotoxin-producing (VTEC) | Diarrheagenic E.coli (1) - hemorrhagic colitis - hemolytic uremic syndrome HUS; most common -> renal failure in peds; maybe fatal esp in young & old - undercooked meat, raw milk, apple cider - toxins vero- or shiga - GS: RBC, no WBC |
| Enterohemorrhagic (EHEC) (2) aka Shiga toxin-producing (STEC) or verotoxin-producing (VTEC) | Diarrheagenic E.coli (2) - E.coli 0157:H7 is most common from bloody stools - non-0157 STEC also causes disease; DNA probe for genes that code for toxins - report to public health |
| Enterotoxigenic (ETEC) | Diarrheagenic E.coli - traveler's diarrhea, diarrhea in infants - contam food or water - toxins - no WBS or RBC - profuse, watery stool - DNA probes to check for toxin genes |
| Enteroinvasive (EIEC) | Diarrheagenic E.coli - bloody diarrhea, dysentery-like, usually kids in poor sanitation - contam food or water - invasiveness - WBC, RBC, mucus |
| Enteropathogenic (EPEC) | Diarrheagenic E.coli - in infants - maj pathogen in developing countries - fec-contam formula & food - adherence-attachement - no WBC or RBC - watery w/ mucus |
| Enteroaggregative (EAEC) | Diarrheagenic E.coli - diarrhea in developing countries; chronic in HIV pts - nosocomial & comm acq - adherence-attachment - most labs can't detect |
| Diffusely adherent (DAEC) | Diarrheagenic E.coli - diarrhea & UTI - most common in kids in developing countries - little known about epidemiology - adherence-attachment - not well studied |
| Characteristics of Nonfermenting Gram-Neg Rods (1) | - obligate anaerobes - no ferm carbs - K/K on TSI - maybe oxidizers or nonoxidizers (asacchrolytic) - oxidation-ferm (OF) medium: either open tube pos/closed tube neg (oxidizer) or open tube neg/closed tube neg (nonoxidizer) |
| Characteristics of Nonfermenting Gram-Neg Rods (2) | - grow on SBA & CHOC in 24-48hrs - most grow on MAC - appear as NLF - most oxidase +; diff from enterobacteriaceae - resist to variety of abx |
| Pseudomonas aeruginosa (1) | Common Nonferm GNR (1) - no normal flora - nosocomial infections: burn, wound RTI, UTI, bacteremia, swimmer's ear, contact lens keratitis - long, thin, pale-staining GNR, slightly pointed or rounded ends |
| Pseudomonas aeruginosa (2) | Common Nonferm GNR - SBA: flat spreading, usually beta hem, dull gray/blue-green, metallic sheen - grows on MAC & EMB (lac neg) - oxidase & catalase +, motile, 42C, grape-like odor, only nonferm w/pyoverdin (fluorescent pigment) - resist to many abx |
| Acinetobacter spp (1) | Common Nonferm GNR (1) - normal skin & pharynx, opportunistic, nosocomial infect (UTI, pneum, septicemia, meningitis, 2nd to P.aeruginosa in freq, A.baumanni most common - pleomorphic GNCB, singles, pairs, short chains, can confuse w/N.gonor,Moraxella |
| Acinetobacter spp (2) | Common Nonferm GNR (2) - may hold crystal violet in broths & smears; confuse w/GPC - most media, MAC: maybe some purplish colonies; confuse w/ LF - oxidase neg (diff from N.gonn), catalase +, nonmotile, resist many abx |
| Stenotrophomonas maltophilia (1) | Common Nonferm GNR (1) - immunocompromised & cystic fibrosis, common in hospitals, nosocomial infec (pneumonia) - straight or slightly curved slender GNR, singles, pairs - SBA: large, nonhem, maybe light yellow, agar maybe lav-green w/heavy growth |
| Stenotrophomonas maltophilia (2) | Common Nonferm GNR (2) - oxidase neg, catalase +, motile, rapid oxidation of maltose, weaker oxidation of glucose, NH4 odor, resist many abx, disk diffusion can be false broth dilution recommended |
| C. jejuni | Campylobacter & Helicobacter - most common diarr; chicks, raw milk, pets - curved slender GNR; "seagulls," loose spirals, S-shaped; faint stain - microaero&capnophilic, CampyBAP 42C CO2, 3 days - darting corkscrew; oxidase, catalase & hippurate hyd + |
| C. coli | Campylobacter & Helicobacter - less severe than C.jejuni, usually food - GS & CX same as C.jejuni - rarely diff from C.jejuni - hippurate hydrolysis neg |
| C. fetus | Campylobacter & Helicobacter - bacteremia immunocompromised & elderly, uncommon stool isolate - GS same as C.jejuni - iso in BLC, 2 wks; inhib on Campy agar; routine media 37C not 42 - oxidase & catalase +, hippurate hydrolysis neg |
| H. pylori | Campylobacter & Helicobacter - gastritis, duodenal & peptic ulcers, poss risk for gastric carcinoma - curved slender GNR - nonsel media 37C, same atm as Campy, not 42C, slow growing - rarely cx, rapid urease test on gastric biopsy, PCR, serology |
| Vibrio cholerae (1) | Vibrio & related (1) - cholera, contam H2O seafood, uncommon in US maybe coastal - small comma-shaped GNR in smears, straight pleomorphic GNR in cx - nonhalophilic, grows on SBA, CHOC, MAC (NLF), lg yellow on TCBS (ferm sucr), APW for enrichment |
| Vibrio cholerae (2) | Vibrio & related (2) - "rice water" stool - oxidase +, motile, serological ID w/antisera to O AG. 01 & 0139 strains cause epidemic, El Tor biotype causes most cases, notify public health, send for confirm |
| Vibrio vulnificus | Vibrio & related - 2nd most serious; immunocomp or w/liver disease: septicemia from raw oysters, water-assoc wound infect; healthy: gastroenteritis - straight or curved GNR - halophilic (add Na+); green or yellow on TCBS; some lac+ - Oxidase+, motile |
| Vibrio parahaemolyticus | - 2nd most common, gastroenteritis, contam seafood, H2O-assoc wound infect, in US - straight or curved GNR - 1% NaCl; SBA, MAC (NLF), blue-green on TCBS (no ferm sucr) - oxidase+, motile |
| Aeromonas spp | Vibrio & related - gastroenteritis, wound infec (aquatic expos), septicemia, meningitis - straight or curved GNR - routine media, most beta hem on SBA, NLF on MAC, no TCBS, CIN & APW for select - oxidase+ (diff from enterobacteriaceae), motile |
| Plesiomonas shigelloides (now included in Enterobacteriaceae) | Vibrio & related - gastroenteritis contam H2O/seafood; bacteremia & meningitis in immunocompromised & neonates - pleomorphic GNR, singles, pairs, short chains, long filaments - SBA, CHOC, most on MAC (NLF), no TCBS - oxidase+, motile |
| H.influenzae (1) | Haemophilus (1) - normal URT; sinusitis, otitis media, pneumonia, bronchitis, often elderly or immunocomp; typeB -> pneumonia, meningitis in kids where no Hib vacc - small, pleomorphic, GNCB - long filaments, maybe capsules |
| H.influenzae (2) | Haemophilus (2) - CHOC in 5-10%CO2, translucent, moist, tannish, encapsulated strains=larger, more mucoid; mousy or bleachy odor; maybe satellitism w/ staph on SBA - should be serotyped |
| H.influenzae biotype aegyptius & H.aegyptius | Haemophilus - conjunctivitis (pink eye) -biotype aegyptius -> Brazilian purpuric fever - characteristics similar to H.influenzae -> difficult to diff |
| H.parainfluenzae, H.haemolyticus, & H.parahaemolyticus | Haemophilus - normal URT, low pathogenicity - small pleomorphic GNCB -> long filaments - large, dry, tannish colonies |
| H.ducreyi | Haemophilus - never normal; chancroid (STD) - small GNCB, bipolar stain, schools of fish or railroad tracks - difficult to cx, can ID by PCR |
| Bartonella | GNR - trench fever, relapsing fever, bacteremia, endocarditis, cat-scratch disease - intracellular GNCB - cx not practical, 9-40 days - dx usually serological or molecular |
| Bordetella pertussis | GNR - whooping cough - small GNCB - Bordet-Gengou & Regan-Lowe media, 3-7 days - fluorescent Ab stain; DTaP vacc |
| Brucella | GNR - Brucellosis/undulant fever; unpast milk/contact w/ goats, cows, hogs, dogs - tiny, faintly staining GNCB - from BLC or biopsy reticuloendothelial tiss; cx not sens - oxid & urease + - L3 pathogen, potential bioterrorism, reportable, dx serology |
| Francisella | GNR - tularemia/rabbit fever; inf by tick or handling animal - pale-staining small pleomorphic intracell GNCB, bipolar staining - media E glu&cys, TM, BCYE - small transparent colonies, 3 days - L3 path, ID direct fluoresc Ab, public health, serology |
| Gardenerella | GNR - normal fem genital, assoc w/BV; neonatal & maternal infec, bacteremia, rarely UTI - small pleomorphic gram-var - no BV dx; SBA, PEA, CNA, HBT, V agar; diffuse beta hem w/ human blood; CO2 2-3d - cat & oxid neg - dx of BV: clue cells, whiff test |
| Legionella | GNR - legionnaire's disease, Pontiac fever; in H2O; inhal of aerosol - small pleomorphic weakly staining - BCYE 3-4d; pale yellow-green fluores w/Wood's lamp - oxidase +, ID w/immunofluorescent stain, serology |
| Pasteurella multocida | GNR - bite/scratch cat/dog or infected carcass; wound & resp infec - pleomorphic GNCB, bipolar staining - SBA & CHOC, 0 MAC; musty odor - oxid, catal, indole + |
| Acceptable | Specimens for Anaerobic Cx - bile, blood, body fluids, bone marrow, percutaneous lung aspirate or biopsy, suprapubic bladder aspirates, tissue, transtracheal aspirate, wound |
| Unacceptable | Specimens for Anaerobic Cx - expec sputum, feces, gastric juice, swabs, voided or cath urine, bronch wash (unless w/ double-lumen plugged cath) |
| Anaerobic Blood agar (CDC) | Media for Anaerobic Cx - nonselective E - obligate & facult anaerobes - yeast extract, L-cysteine, hemin, vit K |
| Bacteroides bile-esculin (BBE) agar | Media for Anaerobic Cx - S, D -> B.fragilis - bile salts & gentamicin (inhibit) - colonies black w/dark halos due to esculin hydrolysis |
| Brucella Blood agar | Media for Anaerobic Cx - E - obligate & facult anaerobes |
| Colistin-nalidixic acid (CNA) blood agar | Media for Anaerobic Cx - S - obligate anaerobes & GP facul anaerobes |
| Cycloserine cefoxitin fructose egg yolk (CCFA) agar | Media for Anaerobic Cx - S, D -> C.difficile - colonies yellow due to fruct ferm; chartreuse fluoresc |
| Egg-yolk agar (EYA) | Media for Anaerobic Cx - determination of lecithinase & lipase production by clostridia & fusobacteria |
| Kanamycin-vancomycin laked blood (KVLB) agar | Media for Anaerobic Cx - aka laked blood kanamycin-vancomycin (LKV) agar - most common S -> anaerobic GNR, esp Bacteroides & Prevotella |
| Phenylethyl alcohol (PEA) agar | Media for Anaerobic Cx - S -| enteric GNR, grows obligate anaerobes & GP facult anaerobes |
| Thioglycolate (THIO) broth | Media for Anaerobic Cx - all-purpose, most things; backup to detect orgs in small #s or anaerobes - thioglycolate = reducing agent - aerobes top, strict anaerobes bottom, facult anaerobes throughout |
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