1. ●BIOCHEMOCAL UNKNOWN ●OXIDATION/ FERMENTATION ●CATALASE ●oXIDASE

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 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

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

8. 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

9. 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