Tube # 11 Lab Report

The following tests according to the lab manual were performed: gram stain, fermentation tubes, methyl red, vogues proskauer, sulfur, indole, motility and growing it up on MacConkey agar. The gram stain was performed incorrectly the first time. This is because the decolorizer was not on the bacterium slide for long enough, giving a false outcome.

There are many differents ways to identify a bacterial unknown and many different situations where identification would be beneficial. One way to identify bacterial unknowns is to perform biochemical tests. In this experiment multiple biochemical tests were done, by performing these tests on the bacterial unknown received the two different bacteria were then identified. The citrate test is done to test the ability of organisms to use citrate as a carbon source. This test uses Simmons citrate agar, the agar contains sodium citrate as the only carbon source and has bromothymol blue as the pH indicator. The organisms that use citrate as a carbon source use the enzyme to transport the citrate into the cell. The cells converts ammonium dihydrogen

A mixed culture of two unknown bacteria was provided by the instructor. The methods used for

I began by running the starch test, which tests for the presence of starch hydrolyzing enzymes. After doing a one-line inoculation of the organism, the plate had to be incubated. Once I received an appropriate amount of growth I added the reagent iodine. The iodine turned the plate purple, formed no clear zone, and lifted the organism off of the plate, which revealed that the starch was not degraded and the enzyme was not present. The organism being lifted off the plate is unique to the bacteria Corynebacterium xerosis indicating that it was my gram positive rod. For reassurance, I ran the Phenol Red Glucose test, which tests if the organism contains various enzymes that determine if the bacteria can ferment glucose. After incubation, the broth turned orange, but this did not provide a clear positive or negative result so I ran the Nitrate Broth Reduction test. The Nitrate Broth Reduction test detects if the organism utilizes nitrate. After incubation for forty-eight hours I added Nitrate A and Nitrate B indicators. However, there was no color change indicating that the test was inconclusive. Since the test was inconclusive, I proceeded to the following step, which included adding a small amount of zinc to the broth, and this turned the broth a red color. The red color indicated that

This test is used to detect if the bacteria contains any deoxyribonuclease activity. Because no color change was observed from blue to clear my unknown bacteria displayed a negative result.

The enzyme urease breaks urea down into NH3 and CO2. An orange broth containing urea is used for this test and needs to be inoculated with the gram negative bacteria. A pink color in the medium indicates a urease-positive organism, an orange or yellow is negative.

A Dichotomous Key was studied to identify bacteria and their relationships. Some of the organisms at the end of the Dichotomous Key had viable characteristics that separate them from different groups, and those that did not students learned how to further classify them. A Dichotomous Key is used to narrow down the search for the unknown organism tested. It is organized by phenotypic characteristics of organisms and conducts a systematic way of identifying the other unknowns. In the lab students were given a tube labeled with a number. Instructions were given to conduct a Gram stain to begin the search followed by the use of a Dichotomous Key and photos as resources to carry out the search. Instructions read to isolate and identify the unknown bacterium with both differential and selective tests to positively identify the given unknown organism. Differential tests used specifically for this unknown microorganism was BEA (Bile Esculin Agar), which interpreted results by the hydrolysis of esculin when the media is blackened around

To perform this test, a tube of broth rich with glucose is acquired. In this tube is phenol red, a pH indicator. Initially, the tube appeared pink in color, indicating a normal pH level. Next, a sample of unknown #44 is introduced into this medium using the aseptic technique, and this is allowed to sit for several days. If the organism is able to ferment glucose, the pH in the medium would decrease and cause the phenol red to exhibit a yellow color. In addition to the straw color, gas can also be produced and trapped inside the Durham tube placed in the medium. This production of acid and gas is a direct result of the fermentation of glucose, as seen with unknown

Preliminary studies help identify Genus species of bacteria. Two different preliminary study pathways must be used since two different pathogens were found in the sample. A dilution and a quadrant streak are the ideal methods to separate pure cultures of bacteria. MacConkey Agar and CAN (MAC) is a selective media that is used for the cultivation of gram negative bacteria. (PEA) is a selective media that is used

In this experiment, an unknown bacterium was given to each individual student. The main purpose of this lab was to identify the given unknown bacteria going through a series of biochemical tests as one of the gram negative bacteria among six different Gram negative bacteria Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa and Salmonella typhimurium. At the very beginning, streaking method; T-streak technique was used to isolate the pure colonies. For the morphological identification of unknown bacteria, Gram Stain Method was done. Biochemical tests that were conducted for the experiment

Two smears of the unknown bacterium #5 were inoculated while the second smear was used for a back up. The unknown bacterium dried for at least forty minutes. After the smears dried, the slides were heat fixed two times to ensure the stability of the organism. The slide was placed on top of the staining rack then over the small sink.

The bases of this experiment was to discover the identify of the unknown from three possible specimens: Klebsiella pneumonia, Escherichia coli, and Enterobacter aerogenes. Utilizaing the T streak technique, the bacteria was isolated into pure colonies for further study. The Gram Stain method was used to identity the morhphology of the bacteria such as the shape and whether the bacteria was Gram positive or Gram negative. Biochemical test were also used to help identify the unknown bacteria. The biochemical test used was the Triple Sugar Iron Agar, Sulfur Indole Motility test, Methyl Red test, Voges-Proskauer test, Citrate test, Urease test, and the Gelatin test. After observing the morphology of the bacteria using the Gram Stain method and utilizing all the possible biochemical test, the bacteria was identified to be Enterobacter aerogenes.

In class, we were given the task of identifying an unknown bacterium broth culture. After receiving number 69, I went through several tests to figure out what bacterium I received. First, I created a slide from my broth by putting a small amount of the unknown broth on to a clean slide and letting it dry for ten minutes. After this, I stained the slide by applying four reagents in order; crystal violet, grams iodine, decolorizer and safranin. From the stained slide, I discovered that this bacterium was gram-negative, which would determine the next couple of tests I would do to identify my unknown bacterium. I began by streaking for confluent growth from my broth culture onto a TSA plate. From the TSA plate, I aseptically transferred a loop

The bacteria that was contained within Unknown tube #12 is believed to be Pseudomonas aeruginosa, Figure 1. The bacteria tested to be Gram Stain negative, producing a pink, red color retained from the staining process. When the species of bacteria was plated on nutrient media, the cells produced an irregular and spreading configuration as shown in Figure 2. This same plating test provided the margins and elevation, lobate and hilly, respectively. The specimen was stabbed in a Fluid Thioglycollate Medium (FTM) tube using an inoculated loop of the bacteria. The results of this experimentation indicate the type of oxygen requirement of the bacteria. The test found the bacteria to be aerobic as colonies of the bacteria began to form along the top of the FTM tube (Manual 2017).

Each test performed in the lab on theses unknown bacteria have a very specific significance. With each test performed correctly the lab officer is able to move closer towards a proper identification of the unknown bacteria. After performing Gram staining and deciphering if the unknown was gram positive or negative the lab officer was then able to proceed to the next step of identification. The gram positive unknown’s reaction to the catalase test informs the tester of the Genus theyre working with. This indicates which tests to perform next. The MacConkey agar is a selective medium that only allows the growth of gram positive bacteria confirming the results received from the gram staining procedure. The NaCl growth test indicates whether or not the organism is able to