●BIOCHEMOCAL UNKNOWN ●OXIDATION/ FERMENTATION ●CATALASE ●oXIDASE

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: _________________________________________________________

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

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

Bacillus cereus 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

Strep salivarius Strep salivarius 26 26. Strep. salivarius: (-) 23 Staph aureus: (+) 15 Micrococcus luteus : (+)

To test for the presence of Cytochrome C Oxidase 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 Oxidase Test: Chrom. Rgnt Red Chrom. Rgnt Ox Chrom. Rgnt Red Cytochrome bo/ bd

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