They are Escherichia coli, Interacted arrogates, Kielbasa pneumonia, Protests miracles, Pseudonymous erogenous, and Salmonella typographic. I will be using several different types of biochemical testing. I am using this type of testing because it differentiate and sometimes identify microorganisms based on specific biochemical characteristics. A gram stain preparation was done first to confirm that the unknown bacterium was gram negative. From here a TTS slant and a TTS plate (for backup) was inoculated with the bacteria and then incubated at ICC between 24-48 hrs.
Afterwards, they were observed for growth and used to perform a series of eight different tests to identify the unknown. Material and Methods Eight different prepared types of media were provided to use to identify the unknown bacterium. They were a Gelatin Tube, MR.. Tube(Methyl Red), UP Tube(Vogues-Prosperous) Urea Tube, SIMI Tube(Sulfur Indolent Motility), Citrate Slant, THIS Slant (Triple Sugar Iron Agar) and Oxides. Each one of the tubes and slants were inoculated using either the streaking or stabbing technique with one of the colonies from the growth of the unknown bacterium.
The Gelatin tube was incubated for up to a 48 hrs at ICC. It was afterwards place in ICC for about 1 hrs to see if it solidifies. The MR.. Tube was incubated for 48 hrs at ICC. Afterwards 3 drops of Methyl Red reagent was place into the tube to observe for color change. Based on the result oaf color change or not would determine if it was positive=mixed acid fermentation occurred or negative= no mixed fermentation occurred. The UP Tube was incubated for 48 hrs at ICC. Afterwards 15 drops (0. 6 m’) Reagent A and 5 drops (0. M’) Reagent B was added o tube to observe for color change in an hour. If a color change occurred the results would be positive=2,3-butadiene fermentation (action produced) or negative=no 2,3-butadiene fermentation (action is not produced). The Urea Tube was incubated for 24 hrs at ICC. Afterwards it was observed for color change. A positive result would have a color change of pink-rapid urea hydrolysis, strong areas production and orange or yellow-no urea hydrolysis, organism does not produce areas or cannot live in broth. The SIMI Tube was incubated for 24-48 hrs at ICC.
Afterwards it was observed for spreading from the stab line, and black precipitate in the medium. Next Kvass Reagent was added to tube and after a minute observed for the formation of red color in the reagent layer. The results for Sulfur present would be positive if black in the medium=sulfur reduction (HAS Production), negative if no black in medium=Sulfur is not reduced. For Indolent present the results would be positive if red in the alcohol layer of Kava’s Reagent= thyrotrophic is broken down into indolent and private, and negative if Reagent color is unchanged=
Tarrytown is not broken down into indolent and private. The result for Motility would be positive if growth radiating outward from the stab line= motility, and negative if no radiating growth=Mennonite. The Citrate Slant was incubated for 48 hrs at ICC . Afterward, it was observed for color change and growth. A positive result would be a blue color change-citrate is utilized and no color change or growth would also indicate positive results. A negative result would be no color change no growth=citrate is not utilized. The THIS Tube was inoculated for 18-24 hrs at ICC.
Afterwards, it was observed for color change and gas production. A yellow slant/yellow butt would indicate glucose and lactose and/ or sucrose fermentation with acid accumulation in slant and butt, Red slant/ yellow butt would indicate glucose fermentation with acid production. Potent categorized aerobically in the slant with alkaline products. Red slant/no change in butt would indicate no fermentation. Potent categorized aerobically with alkaline products. No change in slant ‘any change in but results would be organism is growing slowly or not at all.
Black precipitate in the agar results would be sulfur reduction. Cracks in or lifting of agar results would be gas production. Oxidize Test was performed by placing on a piece of filter paper a loop of the bacterium and then adding a few drops of Oxidize Test Reagent. A positive test would indicate a black color change and negative no color change. Results Gelatin Negative MR.. Positive UP Negative Urea Negative SIMI Eng Sulfur, Positive Indolent, Positive Motility Citrate Negative TAIGAS/A yellow slant/yellow butt; G-Gas production Oxides Negative Discussion/Conclusion
Once all tests were concluded and the results were obtained I used my results to compare to the six given possible organism results to identify the unknown. My test results matched up with Escherichia coli. Escherichia coli is defined as a germ, or bacterium that lives in the digestive tract of humans and animals. There are many types of E. COli. Some types may cause more serious side effects but most are harmless. Most people are diagnosed with E. Coli 0157:HE which sometimes may cause blood problems or kidney failure which can lead to death. Other types of E.