Gram Stain Lab Report

The colonies were smooth, translucent, and had a white brownish color. The Gram stain resulted in Gram positive cocci. After the Gram stain was completed, the bacteria were streaked on a Mannitol-Salt Agar plate and a Catalase test was performed. After these test were completed a Phenol Red Dextrose Fermentation tube was inoculated, and a SIM Tube inoculated.

The purpose of this study project was to carefully isolate and identify two unknown bacteria from a mixed culture. The ability to properly evaluate biochemical test results is also necessary for the identification to be successful. The goal was to apply all of the methods and techniques that have been learned in the microbiology laboratory course for the proper identification of unknown bacteria. A certain amount of bacteria that were used throughout the course were possible bacteria that could be found in a mixed culture. The bacteria that were identified in the mixed culture were Staphylococcus Aureus and Kocuria Rhizophila.

The next step of the project included preparing a Gram stain to discover the cell shape, arrangement, and if the bacteria is gram positive or

The first result of importance was the result of the Gram stain. The observations of the unknown bacteria from the slant culture after Gram staining showed that the unknown bacteria were Gram negative bacilli (Image 1). After determining the unknown bacteria was Gram negative, an oxidase test was conducted on a sample from the slant culture. The cotton swap with the sample of bacteria did not change color when the oxidase reagent was applied, thus providing a negative result. With a negative oxidase test, further tests were conducted to determine various characteristics of the unknown bacteria. A MR-VP broth was inoculated with a sample from a slant culture of unknown bacteria. After incubation, the methyl red reagent was added to the broth, and the broth turned red, providing a positive result (Image 2). An EMB agar streak plate was inoculated with a sample from a slant culture of the unknown bacteria, and after incubation, growth was found on the plate, providing a positive result (Image 3). A Citrate agar slant was inoculated, and after incubation, growth was found on the media, providing a positive result (Image 4). A Urea agar slant was inoculated, and after incubation, the agar had changed from a peach color to a bright pink color, providing a positive result (Image 5). Using the flowchart (Figure 1) developed from the Table of Expected Results, the lab partners started at the oxidase test. Given the negative result of the oxidase test, the flowchart is

In a laboratory setting, it often becomes necessary to identify an unknown organism. In this experiment, researchers classified an unidentified bacterium based on its physical structure, colony morphology, optimal conditions and metabolic properties. A Gram stain using crystal violet, iodine, and safranin and a simple stain using methylene blue characterized the organism’s cell wall. Cultural behavior was classified by inoculating the organism onto nutrient agar and incubating it at 37° C for 48 hours, and observing its behavior, as well as using SIM medium to test for motility. Optimal growth temperature was

The purpose of this lab was to identify two unknown bacteria cultures using various differential tests. The identification of these unknown cultures was accomplished by separating and differentiating possible

The first step toward identifying this unknown organism was to perform a Gram Stain to differentiate between gram positive and gram negative bacteria. This is an important step because it directs what the next tests will be. My Gram Stain on sample #12 showed that the bacteria was gram negative, however, after receiving the results of the OF glucose, H2S, Citrate, Urease and Motility tests, it was apparent that my Gram Stain was contaminated. I then performed a catalase test which came back negative, so I ordered a Bacitracin disc, Optochin disc and a CAMP test which had to be incubated overnight. After receipt of those test results,

Bacteria are ubiquitous; they can be found on the skin, in the soil, and inside the body. Because of the very nature of this ubiquity, it is important to be able to determine between different strains of bacteria. An example of this is determining the causative agent for a disease so that the patient will be treated with the appropriate antibiotics. It may be important to determine the bacteria in a certain region, because like with enteric bacteria, it is normal to find them in the digestive tract as they are in a symbiotic relationship with our bodies in this area; however, they also cause opportunistic infections in places outside of the digestive tract to our detriment, such as with a urinary tract infection. Some strains of bacteria are common to nosocomial infections, and identifying these bacteria as such helps create the guidelines for healthcare workers in antiseptic technique. All of the morphology and characteristics of each strain of bacteria help us to better understand the role of bacteria in the body as well as helps us understand how they can cause illness, and what treatment regimen to set in place. In lab this semester, a sample of unknown

The purpose of this lab was to identify two unknown bacteria cultures using various differential tests. The identification of these unknown cultures was accomplished by separating and differentiating possible bacteria based on specific biochemical characteristics. Whether the tests performed identified specific enzymatic reactions or metabolic pathways, each was used in a way to help recognize those specifics and identify the unknown cultures. The differential tests used to identify the unknown cultures were oxidase, catalase, lactose and sucrose fermentation, Kugler/iron agar, nitrate reduction, gelatin hydrolysis, starch hydrolysis, manitol salt, MR-VP, citrate, bile esculin,

The purpose of this study was to identify the unknown bacterium using biochemical tests and various methods that had been learned from previous the microbiology laboratory class. Identifying the unknown bacterium was determined by separating and differentiating possible

The unknown plate acted as the stock master plate, and was used to aseptically streak the unknown for isolation and was incubated at 37°C for 24 hours. After the incubation, a Gram stain was performed. Using an inoculating loop, a drop of DI water was placed on a clean slide, and a sample of bacteria was aseptically transferred and mixed in the DI water. The slide air dried for a couple of minutes, and was then heat fixed to keep the bacteria on the slide. Using a staining tray to hold the slide, 2 drops of crystal violet dye was placed on the dried bacteria and allowed to sit for 2 minutes (2). After the CV was set, DI water was used to rinse off any remaining dye. The smear was covered in Gram’s iodine for 1 minute. After a minute the iodine was rinsed off and the slide was tipped to the side to allow 2-3 drops of ethanol decolorizer to remove any remaining purple coloring. The slide was rinsed again, and the smear was covered with safranin for 30 seconds. After 30 seconds the slide was rinsed off and blotted dry with bibulous paper (2). The smear was then observed under a microscope to determine gram nature and

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.

The best and most accurate way of identifying an unknown microorganism is by sequencing its DNA, but this is very expensive and only used in highly qualified labs. So, the identification of unknown bacteria number 63 was be done by putting the bacteria through numerous laboratory tests. Microorganisms are different among each other by their macroscopic morphology, microscopic morphology, and the unique metabolic processes they use to survive and reproduce. Identifying an unknown microorganism in the laboratory is important because knowledge is gained on the appropriate way to cultivate an organism, how to correctly read the result of a test, and learning about the different characteristics of the bacteria. All of the following tests were done using the best sterile technique and the most new turbid bacterial growth subculture.

After viewing the end result of the Gram Stain, I saw that the Gram positive bacterial cells were stained violet, while the Gram negative cells were stained red. In the mixed culture under the microscopes, I was able to notice that the Gram positive cells, Enterococcus durans, were in clusters of short chains. The Gram negative, Proteus mirabilis, were in rod-shaped forms that were spread across the slide. These cells did not seem to cluster.

A great deal of cultural tests and physiological procedures were performed on the unknown before the conclusion of P. aeruginosa was made. The first test preformed was Gram’s staining which involves the use of crystal violet, Gram’s Iodine, Ethyl Alcohol, and Safranin. Gram staining is known as a differentiating stain, since it can determine the presence or absence of peptidoglycan in the cell wall. The unknown bacteria were Gram stain negative. After that the motility of the cell was tested using a hanging drop slide. The slide was examined, but the results were inconclusive due to the inability to make a clear distinction between motility and Brownian motion. This test was not