Gram Negative Unknown Lab Report Essays

Gram Negative Unknown Lab Report Essays Gram Negative Unknown Lab Report Paper Gram Negative Unknown Lab Report Paper E. Coli is most commonly found in the intestines of warm blooded organisms. Most E. Coli strands are non pathogenic however, there are strands known to cause food poisoning. Introduction There are multiple reasons for identifying unknown bacteria. In research, it is a must to be able to identify unknown bacteria if you are comparing the different microorganisms. The identification of unknown bacteria may ultimately lead to the discovery of a new species because the results of the tests performed on it may not match those of any other. In my case I had a gram negative bacteria. Gram-negative bacteria are colored red or pink when Gram stained. They have an outer membrane containing alphanumerically that is not found in gram- positive bacteria. This outer membrane functions as an added layer of protection, by shielding the bacteria from several antibiotics, dyes, and detergents that would damage the inner membrane or cell wall. In this particular experiment, I was issued an unidentified organism that could be one of six different bacteria. In order to identify it I was required to run a series of tests and do a comparative analysis. I received my bacteria in a test tube and I inoculated a TTS plate using the T-Streak method in order to isolate the bacteria and also to grow bacteria to use for my tests. Materials and Methods The materials I had available to me for use for this experiment were a TTS plate, a TTS slant, a Gelatin Tube, a Methyl Red Tube, a Vogues-Prosperous Tube, a Urea Tube, a SIMI Tube, a Citrate Slant and a THIS slant, a light microscope, an inoculating loop, and inoculating needle, a burner, an apron, and some glass slides. I did a Gram Stain in order to assure that my bacterium was gram-negative. : I UT a drop of distilled water on a microscope slide and inoculated the bacteria in it. I then heat fixed the bacteria by passing it over a flame three times. Then I applied Crystal Violet to the slide for one minute, and rinsed with distilled water. The slide was rinsed with 95% ethanol for five seconds and immediately rinsed with distilled water. Seafaring stain was added to the slide for two minutes followed by a rinsing with distilled water. A light microscope was used at xx magnification to observe the stained slides. I used the test tube of my unknown bacteria and an inoculating loop to inoculate my TTS plate and TTS slant. I used the T-Streak method on the plate by flaming the loop and streaking quadrant one than flaming the loop again and streaking quadrant two and then flaming the loop again and streaking quadrant three. I then flame the loop again and inoculated the slant. I than incubated them at 37 degrees Celsius for 24 hours and used them to complete the other tests. Next I performed the Gelatins Test using a gelatin tube. I first flamed an inoculating needle and then gathered some colonies from my unknown test tube and stabbed the gelatin tube. This is a differential medium that tests an organisms ability to produce an economy called gelatins that will hydrology gelatin. For the Methyl-Red test I inoculated the test tube with bacteria from my TTS plate and incubated the tube at 37 degrees Celsius for 24 hours. After incubation I placed several drops of the pH indicator Methyl-Red into the test tube and observed the color change of either red or yellow. This test is to see whether the bacterium produces glucose from the mixed acid pathway or not. A red result means the bacteria tested positive. In the Vogues-Prosperous test, I inoculated the tube with bacteria from my TTS late and incubated the tube at 37 degrees Celsius for 3 days. After three days I placed some Barrios Reagent A and Barrios Reagent B in the test tube. The color change of red or pink indicates a positive reaction for action which tells you that the organism is a butadiene ferment. For the Areas test, I inoculated my Urea test tube with my unknown bacteria from a TTS plate using and inoculating loop. The Urea tube was then incubated at 37 degrees Celsius for 8 days to observe for a color change. The Urea tests for the ability of a bacteria grown in urea broth produces areas. This medium notations the pH indicator phenol red. If areas is produced the pH of the media will raise thus causing the phenol red to change from yellow to a pink color. I used an inoculating needle to stab the SIMI test tube and then incubated it at 37 degrees Celsius for 24 hours. The SIMI test was used to test whether an organism has the ability to reduce sulfur to hydrogen sulfide. Iron salts in the media reacts with the hydrogen sulfide to form a black precipitate called ferric sulfide. If sulfur can be reduced than a black color will be seen in the tube. This test also sees if an organism is and indolent producer. Indolent producers are bacteria that produce the enzyme transposes which can hydrology thyrotrophic to private, ammonia and indolent. To test for indolent production, Kavas reagent is dropped into the medium. If there is a red color than the bacteria tests positive for indolent production and brown yields a negative result. The third test that the SIMI test is for is motility. If the bacteria is motile then cloudiness will be seen in the medium and if not the bacteria will only be located where it was stabbed. The Citrate Test was used to test for the ability of a bacteria to utilize citrate as sole source of carbon, these bacteria produce citrate permeate which can transport citrate into the cell and make private from it. Bacteria that can utilize the citrate causes the media to become more alkaline. The indicator for this test is biorhythms blue dye which will turn from green to blue if pH is at 7. And above. A blue color change yields a positive result. I inoculated the THIS (Triple Sugar Iron Agar) Slant with bacteria from my TTS plate using an inoculating loop. The medium was then incubated at 37 degrees Celsius and checked after eight hours of incubation and again after 24 hours f incubation. This test is a differential that tests for sugar utilization, gas production and sulfur reduction. It cont ains lactose, SUcrose and glucose and phenol red as an indicator. No color change means that no sugars were fermented. If there is as bubble or splitting in the medium than a gas was produced and a black color signifies hydrogen sulfide production. The media has sodium tessellate as a reducible sulfur and ferrous sulfate as the hydrogen sulfide indicator. Results The Gelatin test was solid after placing the medium in the cold room for 45 minutes thus it was negative for gelatins. The Methyl-Red test turned red after adding methyl red to the medium indicating a positive result, thus indicating that the unknown produced mixed acids from glucose fermentation. The Vogues- Pressure test had no color change, thus indicating that the unknown was not a butadiene fermented. My unknown remained yellow in the Areas test which means it tested negative thus indicating that the unknown was unable to produce areas. In the SIMI test my unknown had negative results for sulfur reduction and motility but tested positive for indolent production. There was no lack color in the medium and there wasnt a cloudiness either however, the medium turned red when Kavas reagent was added indicating the ability for the unknown to produce transposes. For the Citrate test, my unknown remained green which yields a negative result and the inability of the bacteria to utilize citrate as a sole source of carbon. For the THIS slant my unknown had a yellow slant and a yellow but and it had gas bubbles in the butt thus indicating that glucose and lactose fermentation and the production of a gas. It was negative for sulfur production. All of the previous results were than compared to gram negative unknown chart and the only bacteria that had the same results was Escherichia coli. The unknown #3 was thus identified as E. Coli. Conclusion The identity of the unknown bacterium #3, E. Coli, was supported by all of the tests that were performed for this experiment. The search began with the identification of the bacteria as being rod shaped under the light microscope. Each test was used to eliminate the five other possible choices. The negative result of the Gelatin test eliminated Protest miracles and Pseudopodia organisms as possible candidates. The positive Methyl Red result further eliminated Entertainer arrogates. The negative Vogues-Prosperous test result didnt allow me to rule out any of the remaining candidates. The negative Areas test result knocked Kielbasa pneumonia out as a possible candidate. The negative sulfur production of the SIMI test allowed me to lastly mark Salmonella typographic out a possible candidate leaving only E. Coli. The positive indolent test was only characteristic of E. Coli and the motility of the unknown was in concordance with E. Coli. Also my negative Citrate test was only characteristic of the bacteria E. Coli. The THIS slant really just helped me be sure of three other acetate that my unknown couldnt have been. Escherichia coli also known as E. Coli is a bacterium that is prevalent in the gut of warm blooded organisms. There are several types of E. Coli that are a normal part of the human body and have many beneficial factors including the production of vitamin K. E. Coli also helps prevent harmful bacteria or pathogenic bacteria from growing inside the intestine. Most strains of E. Coli are non pathogenic however, some strains are known to cause things such as food poisoning and serious infections. E. Coli has been the cause of death for many. Symptoms of an E. Lie infection include abdominal pain, diarrhea, nausea, vomiting, fever and fatigue. This pathogen is easily spreader and has received a lot of controversy. E. Coli was named for the person who discovered it, German pediatrician Theodore Escherichia. I feel that this lab was very good for students such as me to see what microbiologists do on a daily basis. I really enjoyed this lab experiment and it has caused me to seek to change m y major to microbiology. I think the identification of unknown bacteria is a very important task and really do have deep respect for the people who make this their profession.