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Cultivating anaerobes and Urine sample analysis

Cultivating anaerobes and Urine sample analysis. Lab #6 Medgar Evers College Prof. Santos. five groups of bacteria based on air requirement. 1-obligate aerobe- these bacteria must be in the presence of oxygen Ex; Pseudomonas aeruginosa.

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Cultivating anaerobes and Urine sample analysis

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  1. Cultivating anaerobes and Urine sample analysis Lab #6 Medgar Evers College Prof. Santos

  2. five groups of bacteria based on air requirement 1-obligate aerobe- these bacteria must be in the presence of oxygen Ex; Pseudomonas aeruginosa

  3. 2-obligate anaerobe- these bacteria can’t tolerate the presence of oxygen. They undergo fermentation or anaerobic respiration. They lack 2 key enzymes, catalase and superoxide dismutase. • Catalase breaks down hydrogen peroxide into oxygen and water • Superoxide dismutase breaks down superoxides into oxygen and hydrogen peroxide *hydrogen peroxide and superoxides are by products of aerobic respiration. • Examples; Clostridium and Bacteroides

  4. 3-facultative anaerobes- these bacteria can grow in the presence or absence of oxygen. Ex; E.coli 4- microaerophiles- these bacteria require a lesser amount of oxygen (5 to 10%) Usually grown in a jar containing a candle! Example; Helicobacter pylori

  5. 5-aerotolerant organisms- these bacteria can grow in the presence of oxygen but do not use oxygen as the final electron acceptor. They usually carry out fermentation, grow better with less oxygen and a higher [ ] of carbon dioxide and water vapor. They lack the enzyme catalase. Example; members of the genus Streptococcus

  6. AIM exercise 21 1- use TGYA shakes to determine the oxygen requirement of certain organisms These contain tryptone, glucose, yeast extract, and agar. They are inoculated while still liquid but cooled to about 450C. They will solidify and the organism will grow where the proper amount of oxygen it needs is!

  7. 2- FTM (fluid thioglycollate medium) containing glucose, cystine, and sodium thioglycollate. Very important, there is a dye called resazurin that turns pink in the presence of oxygen. The tube will appear pink on top since oxygen level is higher on top. There is also some agar that will help localize the anaerobes on the bottom. • This medium supports the growth of both aerobic and anaerobic organisms!

  8. 3- Brewer’s anaerobic agar will be used to cultivate anaerobes! Know the setup for the gaspak anaerobic jar. The important things to remember are; • The gaspak envelope provides hydrogen gas that reacts with the oxygen in the jar to form water and carbon dioxide. This removes the oxygen providing an anaerobic condition. • An indicator strip is placed to tell us when there is no oxygen. The methylene blue strip should become colorless. • The lids should be tightly sealed.

  9. AIM exercise 26 • As far as exercise 26 is concerned, know E.coli is by far the most common cause of urinary tract infections. • A urine sample is examined for color, odor, turbidity, pH, mucus, blood or pus. All these might indicate infection. • You will transfer 0.01 ml of a urine sample and then streaked it on a plate. The following week, you counted the colonies and then after multiplying by 100 one can determine the original count.

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