Plate B incubated at 25oC produced a white, opaque, waxy, and spreading bacteria colonies while plate A incubated at 37oC produced a white, thin spreading bacteria colonies (Figure 1.). Gram staining revealed that plate B contained a rod-shaped bacterium having partial pink and purple coloration with endospores. This suggests that the bacteria is a gram positive bacteria. Gram positive bacteria typically have a thick cell wall which traps the crystal violet making them appear purple. Conversely, plate A is a rod-shaped bacterium but with pink coloration. This appearance is typical of gram negative bacteria because they have thin cell wall, and the crystal violet penetrates through their wall leaving the bacteria colorless. The pink counterstain enters the thin walls and leaves it pink. The glucose fermentation and gas production test carried out on tube D came out negative. Similarly, the mixed acid fermentation test carried out on tube C came out negative (Table 1.). DISCUSSION AND CONCLUSION …show more content…
This indicates that the mixture may contain B. cereus or B. subtilis from the gram positive biochemical chat given. Since the MR-VP test conducted came out negative, this suggests that the unknown organism in plate B is B. subtilis. Also, the Gram staining test carried out on plate A suggested that the plate contained a rod-shaped gram negative bacteria. The glucose fermentation and gas production tests came out negative. Therefore, A. faecalis and P. aeruginosa are the suspected organisms. Since, the growth on plate A is a colony of a white, thin spreading bacteria, it suggests that the organism is A.
An unknown bacterium was handed out by Dr. Honer. The appropriate tests were prepared and applied. The first procedure that was done was the gram stain. Under a microscope, if the gram stain is purple, the bacterium is gram positive, if the stain is red, it is gram negative. The next test was the fermentation tests for glucose, sucrose and
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 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
Next I performed a KOH test to further confirm that my organism was a Gram-negative species. For the KOH test, I added 3 drops of 10% potassium hydroxide (KOH) to a small drop of distilled water onto a clean microscope slide, transferred a visible clump of organism to the KOH solution using my inoculating loop. I than mixed the cells into the solution using small, circular motions for 60 seconds and then lifted up the loop to look for what appears to be a “stringing” affect which means it’s confirmed that it is gram- negative species. Next, I created a streak plate using nutrient agar so that I could see pure culture of my organism. I aseptically obtained a loop full of my organism and gently inoculated one quarter of the nutrient agar plate by running the loop back and forth across the surface. I then flame sterilized the inoculating loop, allowing it to cool for 10 seconds, and then streaked the organism from quadrant I into quadrant II using a zigzag motion technique. I repeated those steps streaking from quadrant II to quadrant III and then streaking from quadrant III to quadrant IV. Once completed, I put the streak plate in the incubator at 37° for 24-48 hours. 48 hours later, I check my streak plate and it had a lot of growth on it. I was able to determine that the organism was definitely an off white color, opaque. The IV quadrant was the quadrant that best
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
There are many reasons for knowing the identity of microorganisms. The reasons range from knowing the causative agent of a disease in a patient, so as to know how it can be treated, to knowing the correct microorganism to be used for making certain foods or antibiotics. This study was done by applying all of the methods that have been learned so far in the microbiology laboratory class for the identification of unknown bacteria. The identification process can be completed with a series of deferential stains and biochemical tests. Creating a dichotomous key helps to limit the amount of biochemical tests done on an unknown organism and by observation
An unknown bacterium was handed out by the lab instructor. The methods that have been learned so far in identifying bacteria were applied to this unknown. Procedures were followed as stated in the lab manual and biochemical test handouts. The first procedure that was done was a gram stain followed by a streak of the unknown on a TSA plate in order to determine the gram reaction and observe the colony morphology. After that, specific biochemical tests were performed for gram positive, since unknown number five was determined to be gram positive rod. The other tests were performed in this order: Mannitol Salt (MSA) streak, Blood Agar streak, Catalase test, Nitrate Reduction test, and Phenyl
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 purpose of this experiment is to distinguish and indentify an unknown bacterium. There are several tests that can help one eliminate and narrow down the options. The most useful test, and the very first one done, is a gram stain. This test will tell whether the bacterium is gram-positive or gram-negative. After the type of gram stain is identified, the tester has a wide array of differentiating tests at their disposal. Based on the results from these tests, and the numerous others that are available, one can accurately establish the identity of an unknown bacterium.
The sole purpose of this project was to identify an unknown bacteria sample #7. Many tests were carried out to determine what this unknown was. Aside from a microbiology lab, understanding and identifying various organisms are important in disease processes, pharmaceutical arenas, and even in the industrial field. Proper lab techniques, including aseptic technique were used throughout the process of identification.
In addition, the purpose of this lab was to identify an unknown organism by performing a series of morphological and biochemical test. The three different types of test that were performed were the Gram stain test, Catalase test, and Red Blood Cell (RBCs) hemolytic test. Thus, the Gram stain test is used to classify bacteria by gram positive or negative and based on their cell
2. Introduction: Each student was given unknown bacteria and was instructed to perform a variety of experimental tests that would help to identify their bacteria. During the process of identification, the unknown bacteria was added to many different testing medias using aseptic technique. They are as follows: lactose fermentation on eosin methylene blue (EMB), TSI (Triple Sugar Iron agar), Phenol red sucrose, the SIM test, H2S by SIM, IMViC (indole, motility, voges-proskauer, and citrate), Urease (urea broth), PDase (Phenylalanine Deaminase), Lysine Decarboxylase, and Ornithine Decarboxylase. Colonial morphology on EMB was used to
Introduction: Through the conduction of numerous experiments, the identity of two bacterial isolates was determined. The tested specimen was an unknown sample of a mixed culture of two different species of bacteria. The first step that was taken was obtaining a pure culture of each species of bacteria by isolating one species from the other. Once isolation was complete, the isolated cultures were tested using procedures that had been performed during previous lab sessions. A gram stain was performed on the two isolates. The isolate which had tested gram negative was then tested for the presence of cytochrome C and lactose fermentation. For the gram positive isolate, cell shape was determined and a catalase test was performed.