Microbiology 20 Biochemical Unknown Spring 2009

(due May 14
th
)


You should be prepared to turn in your notebook with your biochemical unknown
identification completed after lab on Thursday May 14
th
. Out of a possible 70 points you
will be graded as follows:


Successfully performing the necessary tests to identify your unknown (25 pts):

Gram stain (verify with instructor)
o morphology & arrangement
capsule stain
spore stain (only if Gram-negative)
acid fast stain (only if Gram-negative rod)
motility (via wet mount)
colony characteristics on nutrient agar plate (see pg. 70 of lab manual)
o color (observe color of colonies as well as any changes in color of agar)
o diameter of typical isolated colony
o overall appearance
o margin
o elevation
color, growth characteristics on agar slant at 37
o
C and room temp (pg. 79)
characteristics of broth culture (before mixing culture)
o pellicle?
o turbid?
o flocculent?
o sediment?
*oxidation-fermentation/OF test
*starch hydrolysis test (amylase)
*PR-glucose, PR-lactose, PR-sucrose tests (fermentation)
*methyl red/MR test (fermentation mixed acids)
*Vogues-Proskauer/VP test (fermentation acetoin)
*citrate utilization test
*gelatin hydrolysis test (gelatinase)
*urea hydrolysis test (urease)
*phenylalanine deaminase test
*H
2
S test
*motility-indole-ornithine/MIO test
*nitrate reduction test
*oxidase test (cytochrome C oxidase)
*catalase test

* Perform biochemical tests of unknown as part of Exercises 13 to 17.
Adequately recording the procedures and results of each test (25 pts):

I (or anyone else) should be able to repeat your work using only your notebook
your results should be recorded in color and/or sufficiently described and labeled
o indicate the total magnification for all microscopic images
o it should be clear on what date each test was done and analyzed
o procedures already written out in your notebook can be referred to by page
your notebook entries should be neat, well-organized, and easy to read
o set aside a section of pages in your notebook to keep records of your
unknown


Correctly interpreting your results and identifying your unknown (20 pts):

correctly indicating what each result means (e.g., pink Gram stain =Gram-neg.)
conclusion summarizing your results and the identity of your unknown
o can be in chart, table or list form

**Verify identity of unknown using Bergeys Manual of Determinative Bacteriology**



Possible Unknown Bacterial Species


Gram-negative

Aeromonas hydrophila
Alcaligenes faecalis
Citrobacter freundii
Enterobacter aerogenes
Enterobacter cloacae
Escherichia coli
Klebsiella pneumoniae
Proteus mirabilis
Proteus vulgaris
Pseudomonas aeruginosa
Pseudomonas fluorescens
Serratia marcescens
Shigella flexneri
Gram-positive

Bacillus cereus
Bacillus megaterium
Bacillus subtilis
Enterococcus faecalis
Lactococcus lactis
Micrococcus luteus
Micrococcus roseus
Mycobacterium phlei
Sporosarcina ureae
Staphylococcus aureus
Staphylococcus epidermidis
Streptococcus pyogenes
Streptococcus salivarius

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