You are given an unknown bacteria and you need to figure out what is it through

Question

You are given an unknown bacteria and you need to figure out what is it through a series of testing. You are given a list of possible bacteria that are a possible identity for your unknown bacteria. You have four categories and you need to choose 2 tests from each category. Complete a flow chart that includes all eight tests and our expected results based on your research for each of your three organisms. The flow chart will show at least 24 test results. For each test, at least one of your potential organisms must have a result different from the other two. You are using the test results to conclusively identify your unknown organism so each test must be able to isolate one organism from the other two.

You three bacteria are: Bacillus megaterium, Bacillus subtillis, and Escherichia coli

Category 1 – Morphology and Motility Determination: Motility test (motility agar tube and SIM tube), Morphology determination by gram stain, and Colony morphology/cultural characteristics (solid media slants and nutrient broth)

Category 2 – Oxidation/Fermentation Tests: Oxygen Requirements (FTM media and Brewer’s Anaerobic agar), Sugar fermentation tests, Methyl Red (mixed Acid) test, Voges-Proskauer (3-Hydroxybutanone) test, Catalase test (hydrogen peroxide), Oxidase test (oxidase reaent), and Nitrate Reduction test (nitrate test reagents A and B/zinc dust)

Category 3 – Hydrolysis Tests/Biochemical Tests: Starch Hydrolysis test (starch agar plate and Gram’s iodine), Casein Hydrolysis test (skim milk agar), Fat hydrolysis test (spirit blue agar), Tryptophan Hydrolysis test (tryptone broth and kovac’s reagent), Urea Hydrolysis test (urea slant), Hydrogen sulfide production (kilger’s iron agar slant), Citrate Utilization test (citrate agar slant), Phenylalanine Deamination test (phenylalanine agar), and Gel liquefaction (gelatin tube).

Category 4 – Differential/Selective media: Hektoen agar, blood agar, MacConkey agar, EMB (Eosin Methylene Blue) agar, Mannitol Salts Agar, and Brewer’s anaerobic agar

Explanation / Answer

+ve

characters B.megaterium B.subtilis E.coli gram staining positive positive negative colony characters circular, entire, raised, opaque, white large, flat large circular, greyish white, opaque, smooth vp test +ve +ve -ve methyl red -ve -ve +ve starch hydrolysis

+ve

+ve -ve nitrate reduction -ve +ve +ve blood agar gamma hemolytic beta-hemolytic beta hemolytic macconkey's agar inhibits gram positive bacteria inhibits gram positive bacteria flat pink colonies

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