For the last week we performed different tests in order to figure out our unknown bacteria. These are the tests with our results that we performed:

Hydrolytic (digestive) Enzymes
Starch: negative- the bacteria has no enzymes to use starch
Lipid: positive- the bacteria had a slight clear zone indicating it can break down lipids.  This means the bacteria secretes hydrolytic enzymes called lipases.
Casein: negative- there was no clear zone indicating that the milk protein, casein, was not used.
Gelatin: positive-there was some liquid still present after refrigerating the test tube

Utilization of Carbohydrates
Sucrose, Lactose, Manitol: negative- there is no change in color, there are no bubbles.
Glucose: negative- there is a slight change in color, there are no bubbles present.
Methyl Red Test (Mixed Fermentation): negative- no change in color, no acid end products
Voges-Proskauer Test (Butanediol Fermentation): negative- did not use butanediol fermentation
Citrate Utilization Test: positive- blue color, bacteria did grow, the pH indicator brothymol blue turns blue in an alkaline pH. This means bacteria utilizes citrate.

Respiration Tests
Nitrate Reduction Test: positive- it did not change color because nitrate was converted to ammonia. It did not turn a color and this means that nitrite is not present.

Degradation of Amino Acids
Indole (Tryptophan Degradation) Test: negative- did not turn red, tryptophan was not used as a source of energy.

Miscellaneous Tests
Triple Sugar Iron (TSI) Agar Test: The alkaline slant was red, and the alkaline butt was also red.  This means that none of the sugars were fermented so it is not a member of the enterobacteria.  Also, no carbohydrate was fermented and glucose was not used.
Urea Hydrolysis: negative, the urea remained yellow because it was not used.
Litmus Milk Reactions: Negative, when the bacteria was placed in the litmus milk, no gas, clot, or changed happened. It was an alkaline reaction.
Motility Testing: Positive, the bacteria was in a test tube moved up the test medium after sitting overnight in the incubator, indicating that it was motile.

Selective and/or Differential Media
Blood Agar Plate: Bacteria appeared clear and whitish, indicating there was no lyses which means it was gamma.
Eosin Methylene Blue (EMB) Agar: Negative; this test involves enteric vs. non-enteric bacteria, and which bacteria do and do not ferment lactose.  Our bacteria did not grow or produce a green metallic sheen, which means it was negative.
Mannitol Salt Agar: Negative, our bacteria did not grow.  A positive test would have resulted in yellow growth.
MacConkey Agar: Negative, nothing grew.  This test allows you to isolate Gram negative bacteria.  Crystal violet will inhibit gram positive bacteria.  Also you will know whether or not your bacteria produces lactose.  Gram negative will ferment lactose (it will release acid that will have red colonies).  Only gram negative will grow and acid will change the color to red.
Phenylethyl Alcohol (PEA) Agar: Negative, ours did not grow because it is gram negative.  PEA isolates gram positive by inhibiting gram negative bacteria. 

After completing all of these tests, we have concluded that our unknown bacteria is (alkaline) Pseudomonas fluorescens. We concluded that this was our bacteria by following the chart we received in class. First, we know that the bacteria is gram-negative bacilli (rod shaped). Then, we followed the chart down to where the bacteria is negative for the lactose test and negative for the glucose test. Then, we followed the chart down to where the bacteria is positive for nitrate reduction and then alkaline for the litmus milk test. Therefore, according to the chart our unknown bacteria is Pseudomonas fluorescens as confirmed correct by our professor.