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Gram Stain Lab Report

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Abstract This experiment was carried out to determine the species of the unknown organism. Once a choice of the unknown was made a Gram stain was conducted to determine the gram nature and morphology of the organism which was Gram negative bacilli. Based on those results a citrate utilization test was performed resulting in a positive test. Following the flow chart the next test to conduct was a motility test which also had a positive outcome. Lastly, a glucose fermentation test was conducted to determine the unknown organism. The glucose fermentation test yielded negative results indicating that the unknown was Pseudomonas aeruginosa. Introduction P. aeruginosa is a pathogen of plants and animals, it is free living that can be found in water …show more content…

The first of the four test was a Gram-stain. Once the experiment was completed the slide was then placed under a microscope where it was then categorized as Gram-negative bacilli. Its pink color when viewed through the microscope indicated that the test was Gram-negative and its rod shape indicated the morphology was bacilli. With these results the next test to be conducted was the citrate utilization test. A citrate tube was inoculated with P. aeruginosa and incubated at 37 degrees Celsius at 24 hours. Once the time elapsed the tube was viewed and it changed from its green color to a dark blue color indicating a positive test. Based on these results the next test administrated was the motility test. A inculcating needle containing P. aeruginosa was placed into the motility medium using aseptic technique. The tube was then placed into an incubator at 37 degrees Celsius for 24 hours. After 24 hours passed the test was examined and growth was seen radiating from the stab line which indicated a positive test. Lastly, a glucose fermentation test was conducted, P. aeruginosa was aseptically transferred into a tube of glucose broth using an inoculating loop. The tube was then placed into an incubator at 37 degrees Celsius for 48 hours. Once the time had elapsed the tube appeared pink which indicated a negative test. With the negative result the unknown species was identified as P.

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According to the series of test that my group ran for our unknown specimen, we had a match with the bacteria known as Alcaligenes Faecalis. This bacterium belongs to one of the major group of gram-negative bacteria (Phylum Proteobacteria). Alcaligenes Faecalis (Genus, species) is a rod shaped (bacillus), 0.5-1.2 x 1.0-3.0 µm, round with scalloped margin (colony configuration growth), motile (with one to nine peritrichous flagella), gram-negative, non-fermentative bacteria, obligate aerobic, having oxygen as the principal terminal electron acceptor in the electron transport chain (ETC). We consider we have a match with the species Alcaligenes Faecalis because of the following reasons: Fermentation tests performed (Durham sugars) were negative, which indicate that our bacteria use a different metabolic means for growth (non-fermentative gram-negative bacteria).

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Unknown #10 produced no identifiable macroscopic characteristics as a broth, so the first step was to Gram stain a loopful to determine the microscopic characteristics. Gram staining not only helped identify Unknown #10’s microscopic morphology but it also helped ensure the specimen was a pure culture—no other bacteria were visible when Unknown #10 was Gram stained and observed under the microscope. Unknown #10’s key microscopic morphology was that it was a very small, Gram negative bacillus. Though bacilli can possibly form endospores, no empty white centers were visible which suggested that Unknown #10 was not an endospore forming bacteria. No quick endospore stain was performed to validate this assumption since only one assigned organism was endospore forming and unlike Unknown #10, that organism was Gram positive.

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The purpose of this lab report is to employ a myriad of skills, tools and, methods learned throughout this semester to perform the appropriate tests for the identification of the assigned unknown bacteria. Add more background information here!!! The most important tools and techniques used during this identification include aseptic technique, microscopic examination and, the use of selective and differential media. Aseptic technique is an important tool for microbiologists. It is imperative that aseptic technique is maintained throughout the length of any test to avoid any cross-contamination that may lead to inaccurate results.

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The unknown bacteria was then tested on multiple selective and differential media. Growth was present on the MacConkey Agar and the colonies were the same color as the plate, which told me my bacteria was gram negative and did not ferment lactose. There was no growth on the Mannitol Salt Agar, and this told me the unknown was not salt tolerant and did not

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