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Identification of an Unknown Sample

Introduction

This laboratory test aimed to recognize one unknown species from a diversified culture. The researcher conducted three trials in the laboratory to ascertain the unidentified species. The experimenter’s tests include; Columbia CNA Agar, Eosin Methylene Blue Agar, and the Phenol Red Test (Vega & Dowzicky, 2017). The analyzer realized that the unknown species were the Proteus mirabilis bacteria and the streptococcus pyogenes bacteria. Bubbles were not observed in Columbia CNA Agar Test, indicating that the bacteria are Gram-Negative; catalase was not produced. Colonies with dark halos appeared; the Columbia Agar Test did not break down the red blood cells, indicating that the specimen was gram-positive.

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Bubbles were observed in Eosin Methylene Blue Agar Test, which indicates that the bacteria are Gram-Positive; catalase was produced. Bacteria are categorized into Gram-positive and Gram-negative depending on their vulnerability to staining a chemical die during a standard biochemical technique (Vega & Dowzicky, 2017). The possible results during the Phenol Red Test include; colored colonies, which indicate that the species are Gram-positive because they ferment lactose. The sample in the test tube turned yellow, showing the pH of the species in the test tube dropped because of the acid creation and fermentation of the sugars available in the species. Bubbles were not observed during the Phenol Red Test, indicating that the bacteria are Gram-positive; catalase was not produced. Unknown specimens have been used in this report by the researcher to identify the bacteria of the species.

The tester doing this lab test has sufficient information relating to the procedures used in researching and cultivating microbes. The scientist will identify the unknown specimens by analyzing their sugar fermentation, growth properties, and enzyme production (Vega & Dowzicky, 2017). Identifying the unknown species will help the researcher to identify various bacteria via distinct biochemical characteristics and tests. This is crucial in medicine because identifying unknown bacteria helps treat patients by being aware of the bacteria or species contributing to the origin of the illness. Additionally, being knowledgeable about various bacteria is vital because it enables people to develop more antibiotics that patients will use shortly.

Methodology

The scholar inoculated distilled water with a sole bacterial collection to protect the liquid media by touching a pure protecting colony, immersed the loop into distilled water, and thoroughly mixed the solution. To inoculate an agar state medium, the fieldworker placed a small specimen into the agar plate using a sterile swab. The scientist then lifted the lid from the petri dish to inoculate an agar slate medium (Vega & Dowzicky, 2017). The researcher added 5% of the unknown specimens in distilled water to conduct the Columbia CNA Agar Test. The mixture was thoroughly mixed and heated until it boiled. The experimenter then autoclaved the solution for 15 minutes at 121 degrees Celsius (Vega & Dowzicky, 2017). The solution was then allowed to cool to 50 degrees Celsius.

For the Eosin Methylene Blue Agar Test, the researcher suspended 35.96 grams of the species in 1000 ml of distilled water and mixed the solution until the suspension was uniform. The researcher then heated the mixture until it completely dissolved. The expert did sterilization by autoclaving the mix at 121 degrees Celsius for 15 minutes. The third test that the researcher conducted is the Phenol Red Test. The scholar used an inoculating needle and aseptically protected the 30 grams of the species in a test tube (Vega & Dowzicky, 2017). After that, the scholar incubated the test tube holding the species at a temperature ranging between 35 to 37 degrees Celsius for approximately 18 to 24 hours and checked for color changes.

Results

Bubbles were not observed in Columbia CNA Agar Test, indicating that the bacteria are Gram-Negative; catalase was not produced. Colonies with faint halos appeared; the Columbia Agar Test did not break down the red blood cells, indicating that the specimen was gram-positive. Bubbles were observed in Eosin Methylene Blue Agar Test, which suggests that the bacteria is Gram-Positive; catalase was produced. Colorless colonies also appeared in the test tube, indicating that the specimen is Gram-negative because it did not ferment lactose. Colored colonies observed in Phenol Red Test show that the sample is Gram-positive because it ferments lactose. The piece in the test tube turned yellow, indicating the pH of the species in the test tube dropped because of acid creation due to the sugar fermentation of the species. Bubbles were not observed during the Phenol Red Test, indicating that the bacteria are Gram-positive; catalase was not produced. The species in the test tube did not make any gas from the fermentation of the species, hence indicating that the species in the test tube was an acrogenic organism.

Discussion

Eosin Methylene Blue Agar Test is essential in distinguishing gram-negative and gram-positive bacteria. The bacteria that use oxygen generate the catalase enzyme to respire and protect the microbes from toxic by-products eliminated by oxygen metabolism (Vega & Dowzicky, 2017). Eosin Methylene Blue Agar differentiates and isolates gram destructive enteric bacteria from nonclinical and clinical specimens. The researcher used Antimicrobial Drug Susceptibility Disc Diffusion Assay to determine the antimicrobials inhibiting the growth of the fungi causing infections (Vega & Dowzicky, 2017). Columbia Agar is a very nutritious medium applied to isolate and grow different microorganisms, specifically very fussy bacteria such as pneumococci and streptococci in animal samples. Scholars use Columbia Agar for general purposes; researchers can improve the technique by using pure blood. The Phenol Red Test is used for general purposes in the lab to distinguish gram-negative enteric bacteria. It is also used in determining microorganisms depending on their fermentation reactions.

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The Phenol Red Test has a pH indicator (phenol red), peptone, one carbohydrate (either sucrose, lactose, or glucose), and a Durham tube. The lab tests performed by the learner on the unknown specimens detected that the species were Proteus mirabilis bacteria and streptococcus pyogenes. The species had colorless colonies, Gram-negative, and did not ferment lactose during the Eosin Methylene Blue Agar Test (Vega & Dowzicky, 2017). Colorless colonies also appeared in the test tube, indicating that the specimen is Gram-negative because it did not ferment lactose. The sample in the test tube turned yellow, showing the pH of the species in the test tube dropped because of the acid creation and fermentation of the sugars available in the species. Colored colonies observed in Phenol Red Test show that the sample is Gram-positive because it ferments lactose. The presence of the Phenol Red Test is not critical in retaining cell cultures. Researchers often use the Phenol Red Test as a quick way of detecting unknown species. Health practitioners could use the test results to assess effective medicines when treating patients with unknown species.

Reference

Vega, S., & Dowzicky, M. (2017). Antimicrobial susceptibility among Gram-positive and Gram-negative organisms collected from the Latin American region between 2004 and 2015 as part of the Tigecycline Evaluation and Surveillance Trial. Annals of Clinical Microbiology and Antimicrobials, 16(1).

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