Introduction It is important to know the identity of microorganisms for knowing how bacteria works and how it is structured means knowing how it can affect humans. By knowing which agents are causing a disease, one can determine the correct treatment necessary with persons afflicted with the microbes in question. It is also important in knowing and understanding which microbes are beneficial to a person and important to the human body functions. The purpose of this study was to identify the unknown bacteria by applying all the methods that have been learned so far in the microbiology laboratory class. Materials and Methods An unknown bacterium labeled as letter “C” was handed out by Professor Woolam. All methods that have been learned so far for identifying bacteria have been applied to this unknown. All procedures were followed as stated in the course laboratory manual and also as further elaborated on by Professor Woolam. On 29 May 2017, the first procedure to be done was to streak the unknown out on a nutrient agar plate. This was done to ensure a pure and fresh sample of the unknown for the next week. After inoculation of the nutrient agar plate, it was labeled, sealed, and placed aside for incubation and growth for 7 days. Following the preparation of the inoculated nutrient agar plate, a heat fixed slide was prepared and a Gram stain was performed. Unknown C was determined to be a Gramnegative rod. All of the following tests were performed on this unknown: Procedure Purpose Reagents Results Gram Stain Used to determine the chemical makeup of the cell wall. Starch Hydrolysis Test Used to determine if the unknown bacteria metabolize starch with the presence of enzyme amylase. Used to observe bacterial capsule by distinguishing capsular material from the bacteria cell. Used to visualize bacterial endospores that some bacteria use to survive during hostile conditions. Used to differentiate those bacteria that produces an enzyme catalase from noncatalase producin g bacteria. Used to differentiate enterics based on the ability to reduce sulfur and ferment carbohydrates. Used to determine an antibiotic’s ability to kill the bacteria Capsule Stain Endospore Stain Catalase Test Triple Sugar Iron Differential Agar Antibiotic Resistance Test Crystal Violet, Gram’s iodine, 95% ethyl alcohol, & Safranin. Iodine. Gram negative rods Negative for the enzyme amylase. Congo Red & Crystal Violet. Negative for the presence of capsules. Malachite Green & Safranin. Negative for endospore formations. Hydrogen Peroxide. No bubbles were produced. Bacteria is noncatalase producing. N/A Red slant, yellow butt, no gas. Bacteria metabolizes sugar. Penicillin, Tetracycline, & Streptomycin. Resistant to Penicillin. Discussion/Results After a series of differential tests, a chart of bacteria was referenced to discover the unknown bacteria was E. coli. The Gram Stain resulted in a Gram-negative rod shaped bacteria which was not colonized. All other tests conducted went well except for the Endospore Stain which resulted in a false positive for the presence of endospores. This was determined by observing that all “positive” spores were located outside of the bacterial cells and none were located within them. This observation was brought to the attention by Professor Woolam and was quickly confirmed personally. Escherichia coli are bacteria naturally found in the intestines of humans and animals. According to the Center of Disease Control (CDC), “Although most strai ns of E. coli are harmless, others can make you sick. Some kinds of E. coli can cause diarrhea, while others cause urinary tract infections, respiratory illness and pneumonia, and other illnesses” (CDC). A person can contract E. coli from eating raw or undercooked meat or from encountering animal—particularly cattle— feces. It is not recommended to use antibiotics for the treatment of E. coli infections; instead, sufferers are instructed to rest, stay hydrated, and to abstain from treating with an anti-diarrheal. The body generally clears the bacterial infection on its own within 7-10 days.