167069325-Recall-Ascp-Mirobiology.doc

Recall ASCP ’07- ‘08- Must to know  Needs long chain fatty acid- M. furfur  Gram (-) diplococci; (+) lactose, maltose, dextrose, sucrose= N. lactamica  Scotch tape method- E. vermicularis  No sugar fermented, gram (-) diplococci, Oxidase (+) = Moraxella catarrhalis  RPR next test for Treponemes = FTA-ABS  Rotavirus specimen= stool  Resistant to Optochin= viridans streptococci  Gliding motility- Capnocytophaga sp  Yellow smooth colony on EYA, AFB = M. gordonae  Rapid grower AFB= Arylsulfatase (+)  Virulence factor for B. anthracis= capsule  S. maltophilia difference from other NF gram negative rod = Oxidase (-), Dnase (+)  NFO, not fluoresce, non motile- B. mallei  P. aeruginosa like organism in cystic fibrosis= B. cepacia  H. pylori= gastritis, rapid urease  S. paratyphi= lysine (-), H2S (-)  Cryptospridium stain = Kinyoun AFB  Walking pneumonia, resistant to antibiotics, without cell wall= M. pneumoniae  E. cloacae= indole (-), citrate (+), lysine (-), ornithine (+), xanthine (+)  Butcher’s cut, H2S (+), gram (+) rod, alpha hemolytic, non motile= E. rhusiopathiae  High risk to lab personnel- C. immitis  Motility of Listeria is seen at temp= room temp  Hair perforation test= T. mentagrophytes  Fusiform macroconidia= Fusarium  Vancomysin (S), Penicillin (R)= MRSA  CSF storage after inoculation= 35’C  Bile esculin positive spp = group D strep  Biochemical profile of Moraxella catarrhalis= Dnase (+), tributyrin HOH (+), reduce nitrate, Oxidase (+)  Favic chandelier growth of molds= T. schoenleini  Modified acid fast stain = Nocardia spp  Microbiology QC  Daily = Oxidase, GS, Catalase  Weekly= Antibiotics, Biochem Basic Bacteriology  Sodium hypochlorite (bleach)= spillage d.  NALC = liquefy mucus  BSL III= M. tuberculosis  Autoclave- 121’C at 15lbs/psi , 15-20 mins  HEPA filter, negative pressure - BSC  Pasteurization – milk disinfection  Incineration – waste disposal

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Wayson stain= polar bodies of Y. pestis Negative stain= capsule Gimenez = Rickettsiae Iodine stain = inclusion body with glycogen Wright/ Giemsa stain= Borrelia

GRAM POSITIVE COCCI Gram stain Gram positive cocci

( Novobiocin resistant test = differentiate)CNS Catalase

Staphylococci

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Streptococci or Enterococci

MRSA= 35’C, 2% NaCl BAP, Vancomycin Tx Bile esculin HOH = group D Strep Tube Coagulase test= free and bound Sheep blood agar= Streptococci Optochin, bile solubility = Strep. pneumoniae PYR HOH = S. pyogenes , Enterococci SXT sensitive= Group C, F, G beta strep CAMP and Hippurate HOH = Grp B strep Modified Oxidase, OF test = Micrococcus Catalase = 3% H2O2; Staph and Strep Dnase (+) = clear zone/pink zone ( S. aureus) Vancomycin resistant= Pediococcus and Leuconostoc Dick’s test = scarlet fever Abiotrophia = NVS ( Vit B6) UTI = Staph. saprophyticus, Enterococci Prosthetic heart valve inf.= S. epidermidis # 1 skin, wound infection= Staph. aureus # 1 sore throat = Strep. pyogenes # 1 neonatal meningitis= Strep. agalactiae # 1 bacterial pneumonia, # 1 meningitis, # 1 otitis media = Strep. pneumoniae

 Modified TMA = VCN + trimethoprim \ GRAM POSITIVE BACILLI

CLASSIFICATION: Smith and Brown’s Classification 2. beta streptococcus à Complete (Colorless zone) à S. pyogenes, S. agalactiae, Grps C, F & G

3. Gamma streptococcus à no hemolysis ( no zone) à E. fecalis, E. faecium, S. bovis GRAM NEGATIVE COCCI Gram negative cocci Growth on Thayer-Martin agar Pathogenic Neisseria spp.          

( )

Other Neisseria spp.

Oxidase test = Screen test for Neisseria Catalase test (Superoxol) = 30% H2O2 Sugar Utilization test = Speciate Neisseria Nasopharyngeal swab = Carrier of meningococci SPS = inhibits Neisseria Waterhouse Freidericksen = meningococci Dnase test = Moraxella JEMBEC system = transport for Neisseria BAP and CAP= (+) growth N. meningitidis CAP = (+) growth N. gonorrhea

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Niacin test = Yellow (+) for M. tuberculosis Red color (+) = Nitrate red., Tween 80 HOH, Arylsulfatase test MAC = AIDS mycobacterium TCH susceptible= M. bovis Arylsulfatase test= Rapid growers Mycobacteria 42’C = M. xenopi Swimming pool granuloma= M. marinum Potassium tellurite medium= Corynebacterium Medusa head, bamboo, square end, ground glass=B. anthracis Antibiotic associated enterocolitis- C. difficile Terminal spore= Clos. tetani Lecithinase= Clos. perfringes Gray to black colonies= Corynebacterium Double zone hemolysis= theta and alpha toxin Botulinum toxin = canned goods 10u Penicillin disk test= B. anthracis Gas gangrene= C. perfringes Blood culture contaminant- P. acne SPS disk test = P. anaerobius Peptone yeast extract glucose (PYG)= anaerobes GLC = analysis of acid prod. by anaerobes BACTEC 9240/9120 = Fluorescence Actinomyces= lumpy jaw Brick red fluorescence= Veillonella, Prevotella KV agar, KVC resistant, 20% bile= B. fragilis Lactobacillus = Catalase (-), Doderlein bacilli

TB complex

Photochromogens

Rapid growers

Gram Negative Bacilli Scotochromogens

Non photochromogens (MAC)

Non photochromogens (MTC)

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Methyl red test opposite reaction with VP test PAD (+), Urease (+), LD (+)= tribe Proteeae Oxidase negative = Enterics Dnase (+) = Serratia Yellow pigment = E. sakazakii E. coli = bacteuria Non motile = Klebsiella, Shigella, Tatumella Sulfanilic acid, alpha napthylamine= NO3-NO2 Campylobacter = microaerophiles Bile lactose media= enterics H2S producer = Salmonella, Proteus, Edward. Arizona, Citrobacter ONPG = slow lactose fermenter

 Salmonella related to Citrobacter  Shigella related to E.coli  Traveller’s diarrhea= Enterotoxigenic E. coli  O129, Oxidase test = Vibrio and Aeromonas  Lysine (-), H2S (-) = S. paratyphi A  Rice water stool, string test (+) = V. cholerae  Green TCBS= V. parahemolyticus  Moeller’s medium = LOA test  Mineral oil on LOA medium= anaerobic  IMViC = E. coli vs Enterobacter, Klebsiella  Urease test= Helicobacter & Campylobacter  Hippurate HOH = C. jejuni  Sorbitol (colorless) on MacConkey= O157:H7  Red (+) = Indole, MR, VP, Urease, NO3-NO2  Blue (+) = Citrate, Acetamide, Acetate,  Malonate Utilization test  Purple (+) = LOA, Hippurate, LIA, Oxidase  Yellow (+) = ONPG, TSI, V. cholerae TCBS Coliforms (Rapid LF)

Salmonella, Shigella, Serratia

Arizona, Citrobacter

Non Fermentative Orgs ( NFO)  Growth on BAP & MacConkey  TSI = K/K  Oxidase test, Motility, O-F test, Pigment  Decarboxylation of amino acid (LOA)  TSI K/K= NFO except Shewanella (H2S)  OF medium = Inc. CHO, Dec. CHON  Oxidase (-) = Stenotrophomonas, Acinetobacter  MacConkey (-)=Eikenella,Kingella,Mora., Flavo  Pyocyanin (+), Grape like = P. aeruginosa  Growth at 42’C = P. aeruginosa, B. pseudomallei  Wrinkled colony= P. stutzeri, B. pseudomallei  Cetrimide agar = P. aeruginosa  Pits agar= Kingella, Eikenella  Oxidase and MacConkey = Acinetobacter and Chryseobacterium  Swimmer’s ear, Burn, Nosocomial = P. aeruginosa  Cystic fibrosis – P. aeruginosa, B. cepacia  Glander’s, Non motile = B. mallei  Melioidosis = B. pseudomallei  Oxidase (-), Dnase (+) = S. maltophilia

P-M-P Group (Proteeae)

O-F test

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Indicator: Bromthymol blue Positive: Acid (Yellow); Negative: No acid (Green)

Metabolism

Tube w/o oil (open)

Tube w/ oil (close

O/F result

Oxidative

Yellow

green

+/-

Fermentative Yellow

yellow

+/+

Non Green Fermentative

green

-/-

PARVOBACTERIA  GRAM NEGATIVE COCCOBACILLUS  FASTIDIOUS ORGANISMS  Haemophilus  Require X (hemin) and V ( NAD)  Satellitism (H. influenzae)  Media of choice: CAP (horse blood)  X and V, Satellitisim = H. influenzae  Cysteine require= F. tularensis (BSL-III)  Thionine and Fuchsin = Brucella  Oxidase, Urease, Motility= Bordetella  (-) BAP, (-) Mac , (+) CAP = Haemophilus  XV and V strips = H. parainfluenzae (V factor)  CAP= CSF  Porphyrin test = X factor require, Red (+), ALA  Motile, Oxidase, Urease (+)= B. bronchiseptica  CO2, H2S, thionine inhibited= B. abortus  NOT inhibited by thionine-fuchsin= B. melitensis  Whooping cough = B. pertusis  Endocarditis, Castaneda = Brucella  Epiglotitis, Meningitis,Otitis media, Pneumo.= Hib  Soft chancre = H. ducreyi  Safety pin, animal bite wound = Pasteurella  Pits agar, human bite wound = Eikenella  Intracellular = Brucella (undulant fever) Francisella(deerfly fever)  Tumbling motility at 25’C = Listeria  Butcher’s cut, H2S (+) , Catalase (+), Gram(+) rod = Erysiphilothrix  BCYE, air conditioner, Pontiac f.= Legionella  Neonatal meningitis, still birth, food poisoning (cheese), septic abortion = Listeria  Jaundice, urine of rat in flood = Leptospirosis  Darkfield microscopy= spirochetes  Hard chancre = Treponema  McCoy = C. trachomatis  NGU, PID, Conjunctivitis, LGV = C. trachomatis  Weil-Felix = Rickettsiae

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Inclusion body = Chlamydia Giemsa/Wright stain = Borrelia ( relapsing fever) Iodine stain = C. trachomatis (glycogen) DFA = Chlamydia antigen Vincent’s angina = Borrelia and Fusobacterium L forms, haverhill fever, rat bite fever, SPS sensitive = S. monoliformis Violet colony- Chromobacterium Whipple’s disease- Trophyrema Granuloma inguinale = Callymatobacterium Oroya fever, infects RBC = B. bacilliformis Cat scratch dse= B. henselae Infects WBC , morulae body= Ehrlichia Wall less, hemadsorption, PAP = M. pneumoniae Fried egg, NGU, urease (+) A7 medium, T strain.= Ureaplasma TWAR strain= Chlamydia pneumonia Tick transmit= Lyme, RMSF, Ehrlichia Chigger = Orientia tsutsugamushi (Proteus Ox-K ) Louse = Epidemic typhus Rat flea = Endemic typhus DFA = Chlamydia antigen Fletcher’s = Leptospira Periodontal = Capnocytophaga Clue cell, HBTA, V agar, 10% KOH test = G. vaginalis Star like colony= Actinobacillus Gliding motility- capnocytophaga

MYCOLOGY  Hyphae and spore – basic structure of fungi  SDA – general culture media  Czapek’s medium – Aspergillus  Germ tube & Chlamydospore= C. albicans  Malassezia furfur – Tinea versicolor  Cell membrane – target of ketoconazole, amphoteracin B  Trichophyton – skin, hair and nails  Hair Baiting test – T. mentagrophytes  Red color and tear drop conidia – T. rubrum  Favic chandelier (antler) – T. schoenleinii  Club shape macroconidia- Epidermophyton  Coenocytic hyphae- Class Phycomycetes or Zygomycetes  Ascospore – Saccharomyces cerevisiae  Niger (birdseed) seed agar (+) , urease (+), phenol oxidase (+), India Ink = C. neoformans  Germ tube & arthrospore-Geotrichum candidum  Arthrospore – Geotrichum, Coccidioides ,  Trichosporon, Aureobasidium  Blastospores – yeast  Pseudohyphae- Candida,Geotrichum, Trichosporon

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Germ tube (+) – C. albicans, C. stellatoidea Swab – unsuitable for fungal culture 10% KOH – direct microscopic, clearing agent Slide culture (microculture) - save media LPCB = common fungal stain PAS = histologic fungal stain Wright/Giemsa- H. capsulatum Dematiaceous – Phaeohyphomycosis agent Dichotomous branching hyphae-Aspergillus Thrush, Diaper rash, Moniliasis- Candida Meningitis, encapsulated yeast-C. neoformans Pink yeast, Urease (+), blastospore- Rhodotorula Fusiform, sickle cell shape- Fusiform Actinomyces- sulfur granules Nocardia – partial acid fast Macroconidia, microconidia- dermatophytes Sputum- common specimen systemic mycoses CHO assimilation- ID of yeast, free of CHO BHIA – dimorphic fungi High risk to lab personnel, spherule with endospore- C. immitis Chloramphenicol and cycloheximide- Inhibitory agent for mycosel/mycobiotic Cigar shape, flowerlike, sleeves, asteroid- S. schenckii Pigeon’s droppings- C. neoformans Guanos, bats, starlings dropping- H. capsulatum Granules, mycetoma- P. boydii Sclerotic, chromomycosis- Fonsecae, Phialophora, Cladosporium Mariner’s wheel, multiple buds- P. brasiliensis Broad based budding yeast- B. dermatitidis Fusiform macroconidia= Fusarium VIRUS Either DNA or RNA Obligate intracellular pathogen Filtrable agent; nanometer Electron microscopy Cell line or tissue culture Not sensitive to antibiotic a nucleic acid genome (RNA or DNA) Capsid - a protective protein coat Envelope - lipid derived from the host cell m. Viral morphology ( electron microscope)  Capsid symmetry o Icosahedral – DNA, some RNA viruses o Helical – RNA viruses o Complex ( Poxvirus)

VIROLOGY 1. Ether resistant – naked virus 2. Ether sensitive – enveloped virus 3. Acid labile - rhinovirus 4. Acid stable - enterovirus 5. Infantile diarrhea - rotavirus 6. Myxoviruses – has H and N 7. Hemagglutination/Hemadsorption 8. Reye’s syndrome- Chickenpox, influenza 9. Bronchiolitis in children - RSV 10. Hepatitis – Yellow fever, Hep B, CMV 11. Pancreatitis – Mumps, Coxsackie B40 virus 12. Reservoir of Togavirus- birds 13. D-S RNA - Reovirus 14. S-S DNA - Parvovirus 15. TZANCK smear – HSV and VZV 16. Latent Infection - Herpesvirus 17. Rubeola virus - measles 18. Rubella virus – german measles 19. SSPE – measles virus 20. PML – JC virus (polyoma) 21. Prion- Spongiform encephalopathy 22. Viroid- RNA (HDV) 23. Human embryonic fibroblast cell- CMV 24. PMKC, MDCK, hen’s egg- Influenzae 25. 5th disease - Parvovirus 26. 4th disease – Duke’s disease 27. Kaposis sarcoma – HHV-8 28. Filoviridae – Marburg and Ebola 29. Coronavirus – SARS 30. Influenzae virus – H5:N1 ( bird’s flu) 31. Use cotton, dacron, nylon swab – virus 32. Delay of 3 days= 4’C (refrigerator) 33. Delay of 4 days= -70’C not -20’C 34. CPE – tissue culture 35. 33’C - rhinovirus 36. Negative stain / gold, silver/ phosphotungstic acid – Electron microscope 37. Roller drum – hold cell culture tube DNA Viruses (HHAPPPy) Herpesvirus 1. All are D-S DNA except Hepadnavirus Parvovirus Adenovirus 2. All are icosahedral except Poxvirus Poxvirus Papovavirus 3. All are enveloped except PAP Parvovirus (Papovavirus, Adenovirus, Parvovirus) 4. All multiply in nucleus except Rule: RNA viruses 1. All are S-S RNA except Reovirus

2. 3. 4. 5. 6.

All are enveloped except PCR - Picornavirus, Calicivirus, Reovirus All are non segmented except ROBA – Reovirus, Orthomyxovirus, Bunyavirus, Arenavirus Generally helical except the positive sense RNA viruses Positive Sense- Call Pico and Flo To Come Right Negative Sense – Pairing OF Rats at Bunny’s Area