Identifying our Unknown... FINALLY!

Today, we identified our unknown bacteria, L-25! The genus is serratia, we're still working on the genus. We also started our research project in which we're testing different essential oils their bacteriocidal effects. We made a bunch of paper discs to be autoclaved, after determining we would use 10 microliters of water and 10 microliters of each oil to saturate the discs to place on the streak plate of streptococcus pneumoniae. 

Streptococcus pneumoniae

We also checked our plates from last week, finding that the anti-bovine solution reacted and formed a precipitate with the hamburger solution and the bovine solution.

Antibodies and Antigens!

Today, We learned about the ELISA tests and Secondary Antibody tests. We performed a test to determine the purity of a meat sample by poking little holes into agar and injecting bovine and hamburger solutions with Anti-Bovine horse, goat, and pig solutions, to see which ones would react. We also did a control with red and green dye. Then we performed the ELISA test used to detect hepatitis and HIV. First, you coat the bottom with an antigen, rinse with a buffer wash, then add a positive antibody solution, negative antibody solution, and other samples. After washing, add a secondary antibody solution, let sit, rinse, then add a substrate that will change color to indicate a positive test. It was really cool to see tests that are used to detect such serious viruses. We also smelled essential oils that we will be utilizing in our research projects. We also saw that our bacteria that was stored in the freezer had grown when inoculated on an agar plate! 

Yum Yum Yogurt!

Today, we ate the yogurt we made on Tuesday! It was interesting, with the probiotic tablets making a yeasty tasting yogurt and the kefir making a sour cream like yogurt. Then we checked our UV plates, finding that our unknown bacteria, L 25, was not affected by 30 seconds under UV-C light. We made a little fieldtrip to the greenhouse, then got our environmental samples out of the -80 degree freezer. We prepared a streak plate from the environmental samples and put the little tubes back in the freezer. Then we watched a movie on biological terrorism and how scientists in the U.S. traced the anthrax poisonings in 2001.

Yogurt-Makin' :D

Today, we prepared yogurt as a class using Vitamin D milk, Kefir, and Mrs. Pathakamuri's homemade probiotic tablet yogurt! First, we microwaved the milk and let it cool to 37 degrees Celsius. Then we poured the milk into three different labeled cups, Kefir, Yogurt, and Control. We added about a teaspoon of Kefir and yogurt to their respective cups, then covered and put the cups in the incubator at 37 degrees. Tomorrow, the cups will be moved to the fridge and Thursday, WE EAT! We also prepared smear plates of our unknown bacteria and subjected half of the plate to long-wave UV-C light for 30 seconds. After that, we inoculated a beaker of water with unknown bacteria and sterilized it with Dr. Pathakamuri's UV SteriPEN! Today's lab had really practical applications to the real world, I'm so excited to learn more about microbiology and pathophysiology next semester. 

More Results!

Last week, we observed the results of the test tubes we inoculated. Our unknown, L 25, tested positive for Citrate utilization, meaning it converts citrate to citrite to ammonia! L 25 also tested positive for the Indole test, meaning to produces indole as a byproduct by utilizing tryptophan. L 25 tested positive for oxidase test, changing the swab instantly purple like Sarah Bartnicki's hair! L 25, an amazing unknown, was positive for nitrate reduction, because it is a facultative anaerobe, it can reduce nitrate completely into molecular nitrogen!  L 25 tested negative for urea hydrolysis, meaning it does not hydrolyze urea, a molecule used to excrete nitrogen in urine.

Enjoy the pictures, L 25 is almost identified!

Lots of Tests and Antibiotics

Today, we inoculated lots of different test tubes with our unknown sample to determine what enzymes it has and what it utilizes. These tests included the Citrate utilization test, Indole test, Nitrate reduction test, and the Urea hydrolysis test. We also streaked a plate and put 7 different antibiotic chips on to determine which antibiotics are effective. Results Thursday, enjoy the pictures!

EMB, Mannitol, and MacConkey Agar

We streaked EMB, Mannitol, and MacConkey agar plates with our unknown bacteria L 25, and determined that since our bacteria was pinkish on the EMB plate,  it is a gram-negative enteric bacteria, with a lower acid production because it only partially ferments lactose. The mannitol plate was negative, our bacteria does not ferment mannitol. Our MacConkey agar plate confirmed that our gram-negative enteric bacteria partially ferments lactose. It is a facultative anaerobe which oxideses H202, which means it has catalase. Hopefully next week, we will identify it!