1. Microbiology Unknown Lab

2. TSI agar Triple Sugar Iron Agar (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 H 2 S A B C D E 0.1% dextrose 1.0% sucrose 1.0% lactose

3. Penol Red broths ABCDEABCDE A (acid with gas)…..B (acid)……C (uninoculated control)……D (no recation)… …E (alkaline reaction)

4. O-F tests Control E. coli Pseudomonas Alcaligenes Fermenter Oxidizer Nonsaccharolytic

5. Motility Test

6. Decarboxylation Positive Negative uninoculated

7. Gelatin Hydrolysis

8. Simmons Citrate A B C A: Positive…Enterobacter B: Negative…E. coli C: Control-uninoculated

9. Phenylalanine Deaminase Test neg pos

10. Urea agar pos uninoculated neg

11. Urea Hydrolysis Positive uninoculated Negative

12. Methyl Red Test A B A: Positive (E. coli)B: Negative (Enterobacter aerogenes)

13. Voges Proskauer Test A B A: Negative (E. coli)B: Positive (Enterobacter aerogenes)

14. Indole Test Negative Positive - +

15. Catalase Test + - Positive Negative

16. GRAM STAIN

17. 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

18. Test Escherichia coliShigella flexneri Salmonella enteritidis Citrobacter freundi Enterobacter cloacae Enterobacter aerogenes Proteus vulgaris Proteus mirabilis Indol +- or +----+- Methyl Red ++++--++ Voges- Proskauer ----++-- or + Simmons Citrate --+ or -+++- or ++ or - Hydrogen Sulfide --++ or ---++ Urease ---+ or - -++ Motility + or --++++++ Gelatin Hydrolysis ----- or (+) ++ Lysine decarboxylase + or --+--+-- Arginine dihydrolase dd+ or (+)d+--- Ornithine Decarboxylase + or --+- or +++-+ Phenylalanine deaminase ------++ Gas from Glucose +-+++++ or -+ Glucose fermentation ++++++++ Lactose fermentation +--+ or -++-- Sucrose fermentation d--d+++- or + Mannitol fermentation ++ or -++++-- Sorbitol fermentation +- or +++++-- TSI Acid/AcidAlk/Acid Acid/Acid Alk/Acid Or Acid/Acid 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 dmeans different results can be obtained. Tests marked with a d would be unreliable for identification purposes. Differential Identification of Enterobacteriaceae by Biochemical Tests.

19. Test Escherichia coliShigella flexneri Salmonella enteritidis Citrobacter freundi Enterobacter cloacae Enterobacter aerogenes Proteus vulgarisProteus mirabilis Indol +- or +----+- Methyl Red ++++--++ Voges-Proskauer ----++-- or + Simmons Citrate --+ or -+++- or ++ or - Hydrogen Sulfide --++ or ---++ Urease ---+ or - -++ Motility + or --++++++ Gelatin Hydrolysis ----- or (+) ++ Lysine decarboxylase + or --+--+-- Arginine dihydrolase dd+ or (+)d+--- Ornithine Decarboxylase + or --+- or +++-+ Phenylalanine deaminase ------++ Gas from Glucose +-+++++ or -+ Glucose fermentation ++++++++ Lactose fermentation +--+ or -++-- Sucrose fermentation d--d+++- or + Mannitol fermentation ++ or -++++-- Sorbitol fermentation +- or +++++-- TSI Acid/AcidAlk/Acid Acid/Acid Alk/Acid Or Acid/Acid Lets make a dichotomous key ! Where do I start?

20. Methyl Red Enterobacter cloacae Enterobacter aerogenes E.Coli Shigella flexneri Salmonella Citrobacter Proteus vulgaris Proteus mirabilis +- Next…look for a test that distinguishes between the 2 methyl red negtive microbes

21. Test Escherichia coliShigella flexneriSalmonella enteritidisCitrobacter freundi Enterobacter cloacae Enterobacter aerogenes Proteus vulgarisProteus mirabilis Indol +- or +----+- Methyl Red ++++--++ Voges-Proskauer ----++-- or + Simmons Citrate --+ or -+++- or ++ or - Hydrogen Sulfide --++ or ---++ Urease ---+ or - -++ Motility + or --++++++ Gelatin Hydrolysis ----- or (+) ++ Lysine decarboxylase + or --+--+-- Arginine dihydrolase dd+ or (+)d+--- Ornithine Decarboxylase + or --+- or +++-+ Phenylalanine deaminase ------++ Gas from Glucose +-+++++ or -+ Glucose fermentation ++++++++ Lactose fermentation +--+ or -++-- Sucrose fermentation d--d+++- or + Mannitol fermentation ++ or -++++-- Sorbitol fermentation +- or +++++-- TSI Acid/AcidAlk/Acid Acid/Acid Alk/Acid Or Acid/Acid The lysine decarboxylase test distinguishes between these 2 microbes

22. Methyl Red Enterobacter cloacae Enterobacter aerogenes +- NOW…look for a test that distinguishes between the 6 methyl red positive microbes. +- Enterobacter cloacaeEnterobacter aerogenes E.Coli Shigella flexneri Salmonella Citrobacter Proteus vulgaris Proteus mirabilis Lysine decarboxylase

23. 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. Test Escherichia coliShigella flexneriSalmonella enteritidisCitrobacter freundi Enterobacter cloacae Enterobacter aerogenes Proteus vulgarisProteus mirabilis Indol +- or +----+- Methyl Red ++++--++ Voges-Proskauer ----++-- or + Simmons Citrate --+ or -+++- or ++ or - Hydrogen Sulfide --++ or ---++ Urease ---+ or - -++ Motility + or --++++++ Gelatin Hydrolysis ----- or (+) ++ Lysine decarboxylase + or --+--+-- Arginine dihydrolase dd+ or (+)d+--- Ornithine Decarboxylase + or --+- or +++-+ Phenyl alanine deamin ase ------++ Gas from Glucose +-+++++ or -+ Glucose fermentation ++++++++ Lactose fermentation +--+ or -++-- Sucrose fermentation d--d+++- or + Mannitol fermentation ++ or -++++-- Sorbitol fermentation +- or +++++-- TSI Acid/AcidAlk/Acid Acid/Acid Alk/Acid Or Acid/Acid

24. Methyl Red Enterobacter cloacae Enterobacter aerogenes +- +- Enterobacter cloacaeEnterobacter aerogenes E.Coli Shigella flexneri Salmonella Citrobacter Proteus vulgaris Proteus mirabilis Lysine decarboxylasePhenyalanine deaminase 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) +-