BIO225 UNKNOWN 1) Your Names and Emails 2) Unknown Number 3) Identification Process. a) First test: Gram Stain with accurate yet brief description of how that would be performed (again not to wordy but to the point) and Result of gram stain (what all did you learn from the gram stain) Based on the picture that I received of this petri dish, I would classify this as gram-negative. I feel this way because the color is pink, and the Morphology (Shape): Rod (bacillus) shaped and the arrangement primarily single colony. The first test that I would pick is MacConkey to verify gram-negative and rule out gram-positive. By doing this it will narrow down the possible bacteria. The next test I would do would be an Oxidase test to determine if it is aerobic or anaerobic. This would further break down the available options of bacteria by stating if it requires oxygen to grow or not. b) Second Test: etc with brief description and interpretation of results 7/22/2022 The petri dish on the left shows an increasing pH of the media that result in a change in the color of the media to yellow as neutral red turns yellow at alkaline pH. In the right picture: when present, the cytochrome c oxidase oxidizes the test reagent (tetramethyl-p-phenylenediamine) to (indophenols) with an end product that is blue to purple in color. When the enzyme is not present, the reagent remains reduced and is colorless. So Negative Result: No blue color. This indicates that bacteria do not produce cytochrome c oxidase and cannot use oxygen for energy production. So, it’s oxidase negative as well as aerobic because there is growth on the MacConkey test. This is a non-lactose fermenter based on yellow media because there is no red/pink color. I think we should order the Citrate test to further narrow down the organism to see if the organism can reduce nitrate to nitrite. c) Third Test: with brief description and interpretation of results 7/26/2022 Citrate utilization test distinguishes between members of the Enterobacteriaceae family based on their metabolic by-products. This citrate, the test shows that it’s positive citrate because of the blue color. At this moment we are leaning more towards it being Pseudomonas Fluorescens, because it is a gram-negative, aerobic organism, grew on MacConkey, and is Citrate positive. 7/27/2022 d) Fourth test We would like to ask for the Methyl red/MR test and the nitrate reduction test because if they are positive, it would confirm Proteus. If negative it would rule out some more possibilities. f) Fifth test. 7/28/2022 In the beginning, we thought of Pseudomonas, but the Oxidase test from the picture is gray, there is no blue on it. Pseudomonas is oxidase positive, which means the test would’ve shown blue and it did not. Therefore, we’re changing our minds and leaning towards Salmonella or Proteus. Now we are requesting the SIM test and the Urea Hydrolysis test to pick between the 2 possible bacteria. 4) Organism Identity (what do you think your bacterium is) 1) 5) Organism Reservoir 1) 6) Clinical significance of organism (no more than 3 sentences) 7) Which person did what on this report. There is no particular format for this section but is intended to make sure both partners do their part. I have run into this issue in the past and just want to make sure both parties contribute equally. 8) Flowchart showing how you arrived at your answer. (This can be created in any software program and imported into this document or hand drawn then take a picture to insert into this document) Gram stain / \ Gram-positive gram negative | (-) color is pink | Oxidase test / \ Negative Positiven | Colorless (-) | Citrate test / \ Positive negative | positive citrate because of the blue color. Instructions on Performing the Unknown Lab Virtually Finished Above is an example of what your final worksheet might look like once you get enough data from the various tests to identify your bacterium. You might have more tests you might have less tests. The particular bacterium you are randomly assigned will determine the number of tests needed to properly identify the bacterium. Keep in mind that your analysis must be logical without gaps or guessing. What You Submit for a GRADE
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