Ryan Auger BMS 501.

08 Enteric Bacteria Lab Conclusions and Observations Nutrient Agar: Using the non- selective or differential growth media of Nutrient agar we started our Enteric Lab off by viewing the plates prepared before hand. Throughout this Lab we were to complete a variety of test to help determine what random six Genus of bacteria we had from a table provided giving us expected results of the Genus of bacteria. In this first test each plate showed growth of the varying organisms that they were inoculated with. Since, like previously stated this growth media is neither selective nor differential, all the colonies appeared rather similar in appearance other than one. Salmonella, Escherichia, Pseudomonas, Proteus and Shigella all took a semi-translucent appearance, arranged in numerous clusters of abundant growth across the plates. These colonies were mostly circular shaped and ranged in color from a cloudy white to murkier tan. Serratia on the other hand was different in appearance entirely. These colonies were similar in shape and size like all the others, but the colonies were light red in color.

EMB Plates Using the differential media of Levine eosin methylene blue agar (EMB) we were able to determine which bacteria were able to ferment lactose, which was present in the plates. This is observed by growth of pigmented and distinct colonies. Those that can ferment the lactose grow on the plate and produce the pigmented colonies while those that do not ferment lactose may grow, but do not have the same pigmented reaction. Salmonella, Serratia, Pseudomonas, Proteus, and Shigella in the table were

Ryan Auger BMS 501.08 all supposed to have a negative reaction to lactose fermentation, however after talking with you about the growth and color of the these colonies and possible outcomes our group determined that these plates had experienced a partial reaction showing colonies of growth that appeared purplish or pink in appearance. Escherichia had a positive reaction for lactose fermentation and was seen as a colony of a blue-black growth with a metallic green sheen when observed. Additionally this media does not support the growth of gram-positive bacteria.

TSI Agar Using this differential media containing the three carbohydrates glucose, lactose, and sucrose as well as the pH indicator phenol red we are looking to see which of these the bacteria can ferment and produce acid reactions in the inoculated slants. After inoculating the surface (slant) and deeper into the media (butt) where there is no oxygen we looked to determine which bacteria reacted, and in which way. There are three possible reactions in the agar. First if only the glucose is fermented the slant will turn yellow from its original red color, but after time the slant will return back to red. This is due to alkaline conditions reappearing from the digested peptones and a production of ammonium compounds. The second reaction comes from fermentation of lactose or sucrose or both which provides enough acid for the entire slant and but to stay yellow. Lastly if no carbohydrates are fermented by the bacteria the slant will remain red. Salmonella, Serraita, Pseudomonas, and Proteus all remained red in our slants meaning they did not ferment any of the carbohydrates. Escherichia and Shigella both turned the slants and butts yellow meaning they were

Ryan Auger BMS 501.08 able to produce a sufficient amount of acid from fermentation to keep them that way. Additionally there was no gas production seen in any of the tubes noted by the lack of any cracks and fissures possibly due to the age of the media. If present this would indicate the presence of excessive gas production.

IMViC Series In the Indole Test we see which bacteria can digest tryptophan into indole and other products. This is seen by adding Kovacs reagent to the tube of TSA. The presence of a red surface layer indicates indole production. Serratia and Escherichia both showed this layer. Salmonella, Pseudomonas, Proteus, and Shigella showed no reaction.

In the Methyl Red Test we are looking to see the organisms ability to ferment glucose and produce large amounts of acid. This is seen by adding Methyl Red indicator to the tubes. Escherichia turned and remained red indicating a test positive for acid. Pseudomonas, Proteus, and Shigella all turned red on the surface of the tube. Salmonella and Serratia had no color change meaning that there was no fermentation or acid production.

In the Voges-Proskauer Test reagents are added to the tubes to show wether or not the organisms are digesting glucose to acetylmethylcarbinol. If present it will react with alpha naphthol and potassium hydroxide to form a red compound. This only happened in Serratia. All other organisms showed no color change.

Ryan Auger BMS 501.08

In the Citrate Test we were looking to see the ability of certain bacteria to use citrate, a salt of citrate acid, as the only carbon source to grow. When this occurs a color change occurs from blue to deep blue, when the pH of the medium rises and changes the color indicator present in the tube. If citrate does not occur the tube remains the original color. Salmonella and Pseudomonas both had positive results changing the tubes to a deep blue color, while all other organisms remained green showing no ability to use citrate as a growth source.

The Oxidase Test The Oxidase Test is used to see which organisms produce oxidase. We did this by adding hydrogen peroxide to the organisms, which produced a catalase reaction if there was oxidase present. The only organism to have a positive test was Shigella, which was shown by bubble production when the hydrogen peroxide was dropped on it.