Applying the Methyl Red (MR) Test

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The MR test media contains peptone, glucose, and a phosphate buffer (Stout et al, 45). To perform the MR test, I used the stabbing technique to inoculate the MR media. I sterilized the stabbing utensil under an open flame, obtained a small amount of unknown bacteria, and stabbed the MR media. Once the MR media was inoculated, I let it incubate for 24 hours in the 37°C hot room. After the 24-hour incubation, I added 15 drops of Methyl Red to the MR media to test for mixed acid production. VP test (a.k.a. butanediol fermentation test) contains peptone, glucose, and a phosphate buffer in its media, which is the same media as the MR test (Stout et al, 47). To perform the VP test, I used the stabbing technique to inoculate the VP media. I sterilized the stabbing utensil under an open flame, obtained a small amount of unknown bacteria, and stabbed the VP media. Once the VP media was inoculated, I let it incubate for 24 hours in the 37°C hot room. After the 24-hour incubation, I added 15 drops of both Barrits Reagent A and Barrits Reagent B to the VP media, for a total of 30 drops. The Citrate medium, a.k.a. Simmons Citrate Agar, contains sodium citrate as the sole carbon source and ammonium phosphate as the sole nitrogen source (Stout et al, 43). To perform the Citrate test, I used the stabbing technique to inoculate the Citrate media. I sterilized the stabbing utensil under an open flame, obtained a small amount of unknown bacteria, and stabbed the Citrate media. Once the Citrate media was inoculated, I let it incubate for 24 hours in the 37°C hot room. After the 24-hour incubation, I reviewed the Citrate media for a change in the PH of the media, which was indicated by a color change of blue or green. The Urease test has a liqui... ... middle of paper ... ... result of the Gelatin test. The only test that did not match the E. aerogenes identification was the VP test. Possible errors made to not achieve an accurate test could be not waiting a full hour to view the color change or even not using the correct Reagent. TSIA H2S Indole Motility MR VP Citrate Urease Gelatin A/A +g (-) (-) (+) (-) *(-) (+) (-) (-) It is important to know the identification of the E. aerogenes bacteria because it is pathogenic and it is known to cause infections. References: Leboffe, Michael J., and Burton E. Pierce. Microbiology: Laboratory Theory & Application. Englewood, CO: Morton Pub., 2010. Print. Madigan, Michael T. et al. Brock biology of Microorganisms. 13th ed. California: Benjamin Cummings, 2012. Print. Stout, M.A, et al. "Microbiology Lab Notebook". Lab handbook. University of Texas. Arlington. 2014. Print.

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