Microbiology Unknown Lab

TSI agarTriple 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 H2S A B C D E0.1% dextrose

1.0% sucrose

1.0% lactose

Penol Red brothsABCDEA (acid with gas)..B (acid)C (uninoculated control)D (no recation) E (alkaline reaction)

O-F tests Control E. coli Pseudomonas AlcaligenesFermenter Oxidizer Nonsaccharolytic

Motility Test

Decarboxylation Positive Negativeuninoculated

Gelatin Hydrolysis

Simmons Citrate A B CA: PositiveEnterobacterB: NegativeE. coliC: Control-uninoculated

Phenylalanine Deaminase Test neg pos

Urea agar pos uninoculated neg

Urea HydrolysisPositive uninoculated Negative

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

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

Indole TestNegative Positive - +

Catalase Test + -Positive Negative

GRAM STAIN

General Microbiology (BIOL 187) Laboratory Report (Unknown Identification)Page 1: Cover pageInclude your name, the unknown number and you final Genus and species identificationPage 2: Test listingI want a (+) / (-) listing of each test in a vertical, easily scanned format

Page 3: Discussion sectionDetail, in complete sentences, why your results justify your identification and rule out other possibilities.Page 4: About My Microbe sectionResearch 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 keyCreate 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 resultsInclude the handout from lab where your recorded your data as you read the test reactions.Grading:20 pts.Correct Identification10 pts.Discussion section 7 pts."About my microbe section" 8 pts.Dichotomous key 5 pts.Neatness, accuracy and completeness of report50 points total

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 occurrencedmeans different results can be obtained. Tests marked with a d would be unreliable for identification purposes.Differential Identification of Enterobacteriaceae by Biochemical Tests.

TestEscherichia coliShigella flexneriSalmonella enteritidisCitrobacter freundiEnterobactercloacaeEnterobacter aerogenesProteus vulgarisProteus mirabilisIndol+- or +----+-MethylRed++++--++Voges-Proskauer----++-- or +SimmonsCitrate--+ or -+++- or ++ or -Hydrogen Sulfide--++ or ---++Urease---+ or -+ or --++Motility+ or --++++++GelatinHydrolysis----- or (+)- or (+)++Lysinedecarboxylase+ or --+--+--Argininedihydrolasedd+ or (+)d+---OrnithineDecarboxylase+ or --+- or +++-+Phenylalanine deaminase------++Gas fromGlucose+-+++++ or -+Glucosefermentation++++++++Lactosefermentation+--+ or -+ +--Sucrosefermentationd--d+++- or +Mannitolfermentation++ or -++++--Sorbitolfermentation+- or +++++--TSIAcid/AcidAlk/AcidAlk/AcidAcid/AcidAcid/AcidAcid/AcidAcid/AcidAlk/AcidOrAcid/Acid

Lets make a dichotomous key !Where do I start?

TestEscherichia coliShigella flexneriSalmonella enteritidisCitrobacter freundiEnterobactercloacaeEnterobacter aerogenesProteus vulgarisProteus mirabilisIndol+- or +----+-MethylRed++++--++Voges-Proskauer----++-- or +SimmonsCitrate--+ or -+++- or ++ or -Hydrogen Sulfide--++ or ---++Urease---+ or -+ or --++Motility+ or --++++++GelatinHydrolysis----- or (+)- or (+)++Lysinedecarboxylase+ or --+--+--Argininedihydrolasedd+ or (+)d+---OrnithineDecarboxylase+ or --+- or +++-+Phenylalanine deaminase------++Gas fromGlucose+-+++++ or -+Glucosefermentation++++++++Lactosefermentation+--+ or -+ +--Sucrosefermentationd--d+++- or +Mannitolfermentation++ or -++++--Sorbitolfermentation+- or +++++--TSIAcid/AcidAlk/AcidAlk/AcidAcid/AcidAcid/AcidAcid/AcidAcid/AcidAlk/AcidOrAcid/Acid

Methyl RedEnterobacter cloacaeEnterobacter aerogenesE.ColiShigella flexneriSalmonellaCitrobacterProteus vulgarisProteus mirabilis+-Nextlook for a test that distinguishes between the 2 methyl red negtive microbes

The lysine decarboxylase test distinguishes between these 2 microbes

TestEscherichia coliShigella flexneriSalmonella enteritidisCitrobacter freundiEnterobactercloacaeEnterobacter aerogenesProteus vulgarisProteus mirabilisIndol+- or +----+-MethylRed++++--++Voges-Proskauer----++-- or +SimmonsCitrate--+ or -+++- or ++ or -Hydrogen Sulfide--++ or ---++Urease---+ or -+ or --++Motility+ or --++++++GelatinHydrolysis----- or (+)- or (+)++Lysinedecarboxylase+ or --+--+--Argininedihydrolasedd+ or (+)d+---OrnithineDecarboxylase+ or --+- or +++-+Phenylalanine deaminase------++Gas fromGlucose+-+++++ or -+Glucosefermentation++++++++Lactosefermentation+--+ or -+ +--Sucrosefermentationd--d+++- or +Mannitolfermentation++ or -++++--Sorbitolfermentation+- or +++++--TSIAcid/AcidAlk/AcidAlk/AcidAcid/AcidAcid/AcidAcid/AcidAcid/AcidAlk/AcidOrAcid/Acid

Methyl RedEnterobacter cloacaeEnterobacter aerogenes+-NOWlook for a test that distinguishes between the 6 methyl red positive microbes.+-Enterobacter cloacaeEnterobacter aerogenesE.ColiShigella flexneriSalmonellaCitrobacterProteus vulgarisProteus mirabilisLysine decarboxylase

The phenylalanine deaminase testhas 4 negatives and 2 positives for the 6 microbes that are not yet keyed-outIt would be a possibility for the next choice.

TestEscherichia coliShigella flexneriSalmonella enteritidisCitrobacter freundiEnterobactercloacaeEnterobacter aerogenesProteus vulgarisProteus mirabilisIndol+- or +----+-MethylRed++++--++Voges-Proskauer----++-- or +SimmonsCitrate--+ or -+++- or ++ or -Hydrogen Sulfide--++ or ---++Urease---+ or -+ or --++Motility+ or --++++++GelatinHydrolysis----- or (+)- or (+)++Lysinedecarboxylase+ or --+--+--Argininedihydrolasedd+ or (+)d+---OrnithineDecarboxylase+ or --+- or +++-+Phenylalanine deaminase------++Gas fromGlucose+-+++++ or -+Glucosefermentation++++++++Lactosefermentation+--+ or -+ +--Sucrosefermentationd--d+++- or +Mannitolfermentation++ or -++++--Sorbitolfermentation+- or +++++--TSIAcid/AcidAlk/AcidAlk/AcidAcid/AcidAcid/AcidAcid/AcidAcid/AcidAlk/AcidOrAcid/Acid

Methyl RedEnterobacter cloacaeEnterobacter aerogenes+-+-Enterobacter cloacaeEnterobacter aerogenesE.ColiShigella flexneriSalmonellaCitrobacterProteus vulgarisProteus mirabilisLysine decarboxylasePhenyalanine deaminaseE.ColiShigella flexneriSalmonellaCitrobacter

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

Proteus vulgarisProteus mirabilis

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