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Transcript of Unknown Microbe
Enterococcus faecalis Background Mostly found in the large intestine of humans and other animals Known to cause urinary tract infections, bacteremia, endocarditis, meningitis, and other post surgery infections Is feared by healthcare professionals because it is one of the most antibiotic resistant bacteria Although treatment with penicillin and ampicillin is used for urinary infections and wound infections Treatment for endocarditis and meningitis require a combination of penicillin or ampicillin with aminoglycoside Glucose, Lactose, and Sucrose: Bromocresol Purple (pH idicator) Carbohydrate Fermentation Resources Can be transmitted from person to person and also transmitted by food products Enterococci can survive not only in the human large intestine but also in harsh environments, soil, water, food, and plants They can survive 5 day to 4 months on dry lifeless surfaces Mannitol Salt Agar Gram Stain Carbohydrate Fermentation Catalase Test Reagents: crystal violet (primary stain), Gram's iodine (mordant), 95% ethyl alcohol (decolorizing agent), and safranin (counterstain) Products being detected: Differentiation between gram positive and gram negative of the bacterium Gram positive- will stain purple
Gram negative- will stain pink A color change indicates a postitive reaction from acidic waste bioproducts of the microoganisms fermenting these three carbohydrates Bubbles in the broth indicates a positive reaction from CO2 Media: Mannitol salt agar (a high consentrated salt, 7.5% NaCl, phenol red) Mannitol Salt Agar +/- positive growth and did not ferment mannitol
+/+ positive growth and did ferment mannitol
-/- no growth and did not ferment mannitol phenol red is a pH indicator that will turn yellow if the microbe is able to ferment mannital Media: trypticase soy agar Catlase Test this media is selective and differential that inhibits most bacteria other than halophilic microbes A- Color change to yellow and no bubbles
A/G- Color change to yellow and bubbles
Negative- No color change and no bubbles Reagent: 3% hydrogen peroxide Bubbles will form on the slide indicating a positive result The bubbles indicate the microorganism has the enzyme catalase Bubbles do not form on the slide indicating a negative result The absence of bubbles indicate the microorganism does not have the enzyme catalase This test determines if the organism can degrade hydrogen peroxide with the use of the enzyme catalase Gramstain Results Carbohydrate Fermentation (+/-) MSA (+/+) Catalase (-) Gram Positive Cocci Color change in all three tubes and no gas formation: Microbe ferments Lactose, Glucose, and Sucrose Growth and color change: Microbe cna grown on mannitol salt agar and ferments mannitol No bubbles: Microbe does not contain the enzyme catalase http://faculty.sdmiramar.edu/dtrubovitz/micro/dicotkeytouks.html Holt, John, Noel Krieg, Peter Sneath, James Staley, and Stanley Williams. Bergey's Manual of Determinative Bacteriology. 9th. Philadelphia, PA: Lippincott Williams & Wilkins, 2000. 528, 532, 538,552, 553, & 555. Print. <LWW.com>. "Institutional Links." Enterococcus Faecalis. Public Health Agency of Canada, n.d. Web. 08 Oct. 2012. <http://www.phac-aspc.gc.ca/lab-bio/res/psds-ftss/enterococcus-eng.php>. "First Antibiotic In New Class Of Drugs Fights Resistant Infections." FDA Consumer 34.4 (n.d.): 4. Science Reference Center. Web. 8 Oct. 2012. The results of these four main test helped me conclude the best fit possible microbe for the unknown #13 I was given. Summary Once I knew I had a gram positive cocci, I was able to eliminate a huge portion of the other possible microbes. The catalase test narrowed out the Staphylococcus. The fermentation test was a little difficult to determine a definite unknown, so the mannitol salt agar results completed the investigation.