●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

1. Gram Stain: (+)/(-): _________2. Endospore stain: (Spores present/ Spores not present) ___________3.Acid fast stain( Acid fast/ Non acid Fast): _______________________

• Cellular characteristics: ………………………………………………………..6Pts1.Cellular shape: Cocci/ bacilli/ Pleomorphic: ______________________2.Arrangement: Strep (chains)/ Staph (bunch)/ single/ pairs/ tetrads/ other: __________________________________________________________________________________________________________________________

3. Motility(Motile/ Non motile): ______________________ Note: Use the Hanging Drop Method)• Cultural characteristics: ………………………………………………………10Pts

1.Size: ____________________________________________________________2.Pigmentation: _____________________________________________________3.Form: ____________________________________________________________4.Margins: __________________________________________________________5.Elevation: _________________________________________________________

Cont. BIOCHEMICAL UNKNOWN LAB REPORTS

• Biochemical tests: ……………………………………………………………30Pts1.Catalase: _____________2.Oxidase:______________3.Indol: ________________4.MR: _________________5.VP: __________________6.Citrate: _______________7. Urease: ______________8.Phenyl Alanine deaminase: _______________9.Lysine decarboxylase: ___________________10.Ornithine decarboxylase: _________________11.Gelatinase: ____________________________12.H2S production: _________________________

13.Nitrate Reductase: _______________________14.Starch: ______________________________15.Glucose fermentation: __________________16.Lactose fermentation: __________________17.Sucrose fermentation: __________________18.Gas production: ________________________ 19.Mannitol fermentation: __________________20.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 megaterium, Bacillus subtilis, Enterococcus faecalis, Lactobacillus lactis, Micrococcus luteus, Micrococcus roseus, Mycobacterium phlei, Sporosarcina ureae, Staph aureus, Staph epidermidis, Strep pyogenes, Strep salivarius

2626. Strep. salivarius: (-)23 Staph aureus: (+)15 Micrococcus luteus : (+)

Strep salivarius

Strep salivarius

Oxidase Test:a. To test for the presence of Cytochrome C Oxidaseb. To differentiate Neisseria :Cytochrome C oxidase (+)c. To differentiate between E. coli (Cytochrome C Oxidase

(-) from Pseudomonas (Cytochrome C Oxidase (+) d. Take two slidese. Put on top of each slide a piece(2 cm x 2cm) of bibulous

paper.f. With a cotton swab roll a heavy inoculum of E.coli on

one slide and Pseudomonas on the other slideg. Add 2 drops drops of the chromogenic reagenth. Wait for color change

Chrom.Rgnt Red

Chrom.Rgnt Ox

Cytochrome bo/ bd

Chrom.Rgnt Red

OXIDASE TEST

After 5 min of adding the Chromogenic reagent

After 13 min of adding the Chromogenic reagent

Courtesy of Rieko Okawa