Unknown lab report please see picture attached

Materials and Methods:

The first test that took place was to find the desired temperature at which the bacterium will grow. In order to do so, two T-streak TSA plates of Unknown 23B were made and placed in two different temperatures, 25 degrees Celsius and 37 degrees Celsius. They were incubated for 48 hours and then observed for growth. Next, a sample of Unknown 23B was used for gram-staining. A clean slide was applied with water before the sample was smeared. After smearing, the slide was dried in 37 degree Celsius for about 5-10 minutes and heat fixed 3 times. 2 drops of Crystal Violet was applied onto the smear and was left to settle for 2 minutes. The excess stain was washed off by water. Iodine was then applied to the slide and allowed to settle for 1 minute. The slide was then again washed with water and lightly blotted. Acetone-alcohol was applied one drop at a time in 10 second intervals until there was color dispersion before being washed off with water. Afterwards, the slide was flooded with safranin for 60 seconds. The slide was then drained of the excess stain, washed off with water, and lightly dried using a blot booklet. Examination of the stain was done with a microscope, objective oil and 4 objective lenses, which were used to find the arrangement and morphology of the bacterium. A KOH test was also performed on the stain, regardless if it was positive or negative. The test was performed for 30 seconds, reaffirming the results from the gram-stain. After observing the morphology and arrangement of Unknown 33B, a series of tests were completed in order for the family to be identified. The first that was performed was a catalase test.

A TSA plate was streaked with a sample of the Unknown and incubated in 37 degrees Celsius for 48 hours. After incubation, a few drops of H2O2 were placed on a small segment of the streak. Next, a Gas Pak was performed. Two TSA plates were smeared with the Unknown. One of the plates was labeled aerobic and the other one was labeled anaerobic. The plate labeled as anaerobic was sealed in a Gas Pak. Both plates were then incubated in 25 degrees Celsius for 48 hours. The results from the test determined whether they were aerobic or facultative. A motility test was performed to determine the bacterium is capable of movement. Using a motility tube, a broth sample of the Unknown was stabbed into the tube in a straight path. The tube was then incubated at 25 degrees Celsius for 24 hours. The next test that was performed was glucose fermentation. A tube of glucose was inoculated with the unknown. After inoculation, the tube was placed in 37 degrees Celsius for 48 hours. After determining the family of the Unknown, using the Bergey’s Manual, more tests were performed. The first set of tests performed was to determine salt tolerance. Three individual TSB tubes with NaCl percentages was used: 5%, 10%, and 15%. All three tubes were inoculated with the bacterium and incubated in 25 degrees Celsius for 48 hours.