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Microbiology Unknown Lab. 0.1% dextrose 1.0% sucrose 1.0% lactose. TSI agar Triple Sugar Iron Agar. A B C D E. (a) Red/red (no sugar fermentation) (b) Control (c) Red/yellow (Glucose fermented but lactose and sucrose not fermented)

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tsi agar triple sugar iron agar

0.1% dextrose

1.0% sucrose

1.0% lactose

TSI agarTriple Sugar Iron Agar

A B C D E

(a) Red/red (no sugar fermentation)

(b) Control

(c) Red/yellow (Glucose fermented but lactose and sucrose not fermented)

(d) Yellow/yellow (Glucose fermented. Lactose and/or sucrose fermented)

(e) Red/yellow with H2S

penol red broths
Penol Red broths

A B C D E

A (acid with gas)…..B (acid)……C (uninoculated control)……D (no recation)… …E (alkaline reaction)

o f tests
O-F tests

Control E. coli Pseudomonas Alcaligenes

Fermenter Oxidizer Nonsaccharolytic

decarboxylation
Decarboxylation

uninoculated

Positive Negative

simmons citrate
Simmons Citrate

A B C

A: Positive…Enterobacter

B: Negative…E. coli

C: Control-uninoculated

urea hydrolysis
Urea Hydrolysis

Positiveuninoculated Negative

methyl red test
Methyl Red Test

A B

A: Positive (E. coli) B: Negative (Enterobacter aerogenes)

voges proskauer test
Voges Proskauer Test

A B

A: Negative (E. coli) B: Positive (Enterobacter aerogenes)

indole test
Indole Test

Negative Positive

- +

catalase test
Catalase Test

+ -

Positive Negative

slide16

General Microbiology (BIOL 187) Laboratory Report (Unknown Identification)

Page 1: Cover page

Include your name, the unknown number and you final Genus and species identification

Page 2: Test listing

I want a (+) / (-) listing of each test in a vertical, easily scanned format

Page 3: Discussion section

Detail, in complete sentences, why your results justify your identification and rule out other possibilities.

Page 4: About My Microbe section

Research your microbe and give me a several paragraphs about your microbe.

(Don't copy word-for-word from your textbook---This will be frowned upon!)

Page 5: Dichotomous key

Create a neat dichotomous key, which results in the correct separation of all 8 possible unknowns. This key should be legible and sized to occupy a complete sheet of paper (nothing microscopic is acceptable here)

Page 6:Lab handout where you recorded your initial results

Include the handout from lab where your recorded your data as you read the test reactions.

Grading:

20 pts. Correct Identification

10 pts. Discussion section

7 pts. "About my microbe section"

8 pts. Dichotomous key

5 pts. Neatness, accuracy and completeness of report

50 points total

differential identification of enterobacteriaceae by biochemical tests
Differential Identification of Enterobacteriaceae by Biochemical Tests.

How do I interpret the table above??

+ means 90% or more of the strains are positive

- means 90% or more of the strains are negative

+ or - means most strains are positive but a significant number are negative

- or + means most strains are negative but a significant number are positive

(+) means a positive result may be weak or delayed in its occurrence

d means different results can be obtained. Tests marked with a d would be unreliable for identification purposes.

slide19

E.Coli

Shigella flexneri

Salmonella

Citrobacter

Proteus vulgaris

Proteus mirabilis

Enterobacter cloacae

Enterobacter aerogenes

-

+

Methyl Red

Next…look for a test that distinguishes between the 2 methyl red negtive microbes

slide21

E.Coli

Shigella flexneri

Salmonella

Citrobacter

Proteus vulgaris

Proteus mirabilis

Enterobacter cloacae

Enterobacter aerogenes

-

+

Methyl Red

-

+

Lysine decarboxylase

Enterobacter cloacae

Enterobacter aerogenes

NOW…look for a test that distinguishes between the 6 methyl red positive microbes.

slide22

The phenylalanine deaminase test

has 4 negatives and 2 positives for the 6 microbes that are not yet “keyed-out”

It would be a possibility for the next choice.

slide23

E.Coli

Shigella flexneri

Salmonella

Citrobacter

Proteus vulgaris

Proteus mirabilis

Enterobacter cloacae

Enterobacter aerogenes

-

+

Methyl Red

-

+

-

+

Lysine decarboxylase

Phenyalanine deaminase

Enterobacter cloacae

Enterobacter aerogenes

E.Coli

Shigella flexneri

Salmonella

Citrobacter

(these 4 still need to be “keyed-out” from each other)

Proteus vulgaris

Proteus mirabilis

(these 2 still need to be “keyed-out” from each other)