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EXPERIMENT 9( T)

EXPERIMENT 9( T). BIOCHEMICAL UNKNOWN OXIDATION/FERMENTATION CATALASE OXIDASE STARCH HYDROLYSIS UREASE. BIOCHEMICAL UNKNOWN LAB REPORTS

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EXPERIMENT 9( T)

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  1. EXPERIMENT 9( T) • BIOCHEMICAL UNKNOWN • OXIDATION/FERMENTATION • CATALASE • OXIDASE • STARCH HYDROLYSIS • UREASE

  2. BIOCHEMICAL UNKNOWN LAB REPORTS Name_______________________ Unknown # ________ Take an unknown and write down its number. Inoculate 2 nutrient agar slants. Inoculate one agar plate w/ the streak method of isolation of colonies. Study the unknown following this format: • Flow Chart: ………………………………………………………………………5Pts • Staining: ………………………………………………………………………….9Pts • Gram Stain: (+)/(-): _________ • Endospore stain: (Spores present/ Spores not present) ___________ • Acid fast stain( Acid fast/ Non acid Fast): _______________________ • Cellular characteristics: ………………………………………………………..6Pts • Cellular shape: Cocci/ bacilli/ Pleomorphic: ______________________ • Arrangement: Strep (chains)/ Staph (bunch)/ single/ pairs/ tetrads/ other: ______________________________________________________________ • ______________________________________________________________ • 3. Motility(Motile/ Non motile): __________________________ Note: Use the Hanging Drop Method) • Cultural characteristics: ………………………………………………………10Pts • Size: ____________________________________________________________ • Pigmentation: _____________________________________________________ • Form: ____________________________________________________________ • Margins: __________________________________________________________ • Elevation: _________________________________________________________

  3. Cont. BIOCHEMICAL UNKNOWN LAB REPORTS • Biochemical tests: ……………………………………………………………30Pts • Catalase: _____________ • Oxidase:______________ • Indol: ________________ • MR: _________________ • VP: __________________ • Citrate: _______________ • Urease: ______________ • Phenyl Alanine deaminase: _______________ • Lysine decarboxylase: ___________________ • Ornithine decarboxylase: _________________ • Gelatinase: ____________________________ • H2S production: _________________________ • Nitrate Reductase: _______________________ • Starch: ______________________________ • Glucose fermentation: __________________ • Lactose fermentation: __________________ • Sucrose fermentation: __________________ • Gas production: ________________________ • Mannitol fermentation: __________________ • O/F: __________________________________ • Brief literature review about the unknown: ………………………10Pts • Unknown # _____ is: ______________________ Total .……….70Pts

  4. Aeromonas hydrophila, Alcaligenes faecalis, Citrobacter freundii, Enterobacter aerogenes, Enterobacter cloacae, E. coli, Klebsiella pneumoniae, Neisseria sicca, Proteus mirabilis, Proteus vulgaris, Pseudomonas aeruginosa, Pseudomonas fluorescens, Serratia marcescens, Shigella flexnerii

  5. Bacillus cereus, Bacillus megaterium, Bacillus subtilis, Enterococcus faecalis, Lactobacillus lactis, Micrococcus luteus, Micrococcus roseus, Mycobacterium phlei, Sporosarcina ureae, Staph aureus, Staph epidermidis, Strep pyogenes, Strep salivarius

  6. Strep salivarius Strep salivarius 26 Strep.salivarius: (-) Staph aureus: (+) Micrococcus luteus : (+)

  7. Oxidase Test: To test for the presence of Cytochrome C Oxidase To differentiate Neisseria :Cytochrome C oxidase (+) To differentiate between E. coli (Cytochrome C Oxidase (-) from Pseudomonas (Cytochrome C Oxidase (+) Take two slides Put on top of each slide a piece(2 cm x 2cm) of bibulous paper. With a cotton swab roll a heavy inoculum of E.coli on one slide and Pseudomonas on the other slide Add 2 drops drops of the chromogenic reagent Wait for color change Chrom. Rgnt Ox Cytochrome bo/ bd Chrom. Rgnt Red Chrom. Rgnt Red

  8. OXIDASE TEST After 5 min of adding the Chromogenic reagent After 13 min of adding the Chromogenic reagent Courtesy of Rieko Okawa

  9. epi

  10. Urease Test 3

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