As shown in Figure 1, there are purple and pink cells. The purple cells are the gram-positive and the pink cells are the gram-negative bacteria. It shows that the gram-negative are bacillus and are smaller than the gram-positive cells. As shown in Figure 2, the MAC plate selects for gram-negative bacteria. It is observed that they are medium in size, raised, and uniform. It also shows that the bacteria are lactose fermenters because of the pink color. As shown in Figure 3, there are two different bacteria growing on the plate based on the presence of two different size bacterium. Also seen in this figure, the unknown gram-negative is gamma-hemolytic because there is no change behind or around the growth. As shown in Figure 4, the gram-negative …show more content…
A colony from this plate was able to be removed and then used to inoculate a TSA slant which would be used for further testing. The gram stain performed on the mixed culture allowed for the morphology of the bacteria to be determined. The gram-negative bacteria were bacillus and medium in size. The MAC plate containing the mixed culture was selective and only allowed for the growth of the gram-negative bacteria. The MAC plate is also a differential medium that shows whether the bacteria are lactose fermenters or non-lactose fermenters. As shown in Figure 2, the gram-negative bacteria were lactose fermenters so that indicated that the next test to be performed and observed was the MRVP test. As shown in Figure 5 and 6, the gram-negative bacteria were Methyl Red positive and Voges-Proskauer negative. This indicates that the next test to be performed and observed is the citrate test. The result of the citrate test was confirmed by the SIM tube used for the indole test. The development of a black color would indicate that the bacteria was H2S positive which also indicates that the bacteria is citrate positive and vis versa (2). In conclusion, the unknown gram-negative bacteria was determined to be Escherichia coli. This was confirmed by the flow chart provided for the gram-negative isolate. Although the chart indicated that the indole test was not needed to identify the bacteria, it was used to confirm the identity alongside the urease test. As shown in Figure 7 and 9, E.coli have a positive result for the indole test and a negative result for the urease test
I identified the genus and species of an unknown bacterial culture, #16, and I applied the following knowledge of morphologic, cultural and metabolic characteristics of the unknown microorganism according to the laboratory manual as well as my class notes and power point print outs. I was given an incubated agar slant labeled #16 and a rack of different tests to either examine or perform myself; the tests are as follows: Gram Stain; Nutrient Gelatin Test; Carbohydrate Fermentation; Dextrose, Lactose and Sucrose; IMVIC tests; Citrate, Indole, Mythel-Red and Vogues Proskauer test; as well as a Urease and TSI Test. Materials and Methods/Results Upon receiving the Microorganism (M.O.) #16, I prepared a slide by cleaning and drying it. Then, using a bottle of water I placed a sterile drop of water on the slide and used an inoculating loop, flame sterilized, I took a small sample of the unknown growth in my agar slant and smeared it onto the slide in a dime sized circle and then heat fixed it for ten minutes.
The unknown bacterium that was handed out by the professor labeled “E19” was an irregular and raised shaped bacteria with a smooth texture and it had a white creamy color. The slant growth pattern was filiform and there was a turbid growth in the broth. After all the tests were complete and the results were compared the unknown bacterium was defined as Shigella sonnei. The results that narrowed it down the most were the gram stain, the lactose fermentation test, the citrate utilization test and the indole test. The results for each of the tests performed are listed in Table 1.1 below.
The isolate possesses some enzymes required for hydrolytic reactions. Hydrolytic enzymes found to be secreted from the bacterium, are -amylase, casein, and PYRase. In the starch hydrolysis and casein tests, there was a zone of clearing around the bacterium, which was indicative of the secreted enzymes necessary to break down starch and casein. In the PYR test, the presence of PYRase was detected by a color change to red on the PYR disc after the addition of the PYR reagent (p-dimethylaminocinnamaldehyde). Hydrolytic enzymes for which the EI tested negative were urease, gelatinase, and DNAse. In the Urea Hydrolysis test, it was observed that the urea broth did not have a color change, indicating that there was no urease secreted to break down urea in the broth. Similarly, there was no gelatinase present to break down gelatin in the Gelatin Hydrolysis test, so the nutrient gelatin remained solid. It was concluded that the EI does not possess DNase because there was no clearing zone around the bacteria, indicating that DNA had not been
...indole, it is motile, there is no urease present and there is no coagulase activity. By deduction and logical reasoning Unknown 10a was determined to be Escherichia coli.
In this lab project, the microbiology students were given 2 unknown bacteria in a mixed broth each broth being numbered. The goal of this project is to determine the species of bacteria in the broth. They had to separate and isolate the bacteria from the mixed broth and ran numerous tests to identify the unknown bacteria. The significance of identifying an unknown bacteria is in a clinical setting. Determining the exact bacteria in order to prescribe the right treatment for the patient. This project is significant for a microbiology students because it gives necessary skills to them for future careers relating to clinical and research work.
The purpose of this study is to identify an unknown bacterium from a mixed culture, by conducting different biochemical tests. Bacteria are an integral part of our ecosystem. They can be found anywhere and identifying them becomes crucial to understanding their characteristics and their effects on other living things, especially humans. Biochemical testing helps us identify the microorganism present with great accuracy. The tests used in this experiment are rudimentary but are fundamental starting points for tests used in medical labs and helps students attain a better understanding of how tests are conducted in a real lab setting. The first step in this process is to use gram-staining technique to narrow down the unknown bacteria into one of the two big domains; gram-negative and gram-positive. Once the gram type is identified, biochemical tests are conducted to narrow down the specific bacterial species. These biochemical tests are process of elimination that relies on the bacteria’s ability to breakdown certain kinds of food sources, their respiratory abilities and other biochemical conditions found in nature.
The purpose of this laboratory is to learn about cultural, morphological, and biochemical characteristics that are used in identifying bacterial isolates. Besides identifying the unknown culture, students also gain an understanding of the process of identification and the techniques and theory behind the process. Experiments such as gram stain, negative stain, endospore and other important tests in identifying unknown bacteria are performed. Various chemical tests were done and the results were carefully determined to identify the unknown bacteria. First session of lab started of by the selection of an unknown bacterium then inoculations of 2 tryptic soy gar (TSA) slants, 1 nutrient broth (TSB), 1 nutrient gelatin deep, 1 motility
The eighteenth exercise of the laboratory manual titled Unknown Identification and Bergey’s Manual is an experiment to identify an unknown bacterium. In this exercise, a student must randomly choose a numbered bacterium available to the class. The keys in Appendix H, located on the last pages of the book, are the major helpful tools in this exercise because it provides completed steps of tests that needs to be performed in order to distinguish certain bacteria. This means that in this exercise, various types of tests and techniques must be performed to identify the chosen unknown bacterium. The unknown bacterium that I selected was number thirty-nine in which I discovered as the Bacillus megaterium after conducting several tests.
I was given unknown organism #14, in order to find out what organism I had, I had to perform several different biochemical tests to identify it. Starting with the Gram stain test, which is performed to differentiate Gram-positive and Gram-negative cells. After staining, when observed through the microscope Gram-positive cells are a purple color with thick peptidoglycan cell walls. Gram-negative cells are a pinkish/red color with thinner cell walls. (handout G. s.) My organism was observed to be pinkish rod shaped meaning it is Gram-negative bacteria.
In the last decade, the number of prescriptions for antibiotics has increases. Even though, antibiotics are helpful, an excess amount of antibiotics can be dangerous. Quite often antibiotics are wrongly prescribed to cure viruses when they are meant to target bacteria. Antibiotics are a type of medicine that is prone to kill microorganisms, or bacteria. By examining the PBS documentary Hunting the Nightmare Bacteria and the article “U.S. government taps GlaxoSmithKline for New Antibiotics” by Ben Hirschler as well as a few other articles can help depict the problem that is of doctors prescribing antibiotics wrongly or excessively, which can led to becoming harmful to the body.
mutans was problematic due to its difference with Bergey’s Manual result for the catalase test. However, after comparing it with a peers results, it seems very possible that the strain we are working with varies from the strain used in Bergey’s. Bacteria possess the ability to develop varying phenotypes within the same species due to frequent mutation and horizontal gene transfer. Therefore, it is possible that the results obtained in our lab may vary from those provided in Bergey’s Manual. Arriving to the conclusion that the Gram negative bacteria was Klebsiella pneumoniae was much more direct. Using Bergey’s Flowchart for identification, the bacteria shared the test results and had a similar shape and
During this investigation to identify the unknown bacterium, seven different biochemical tests were performed. All biochemical tests were performed according to the Eastfield Microbiology Lab Manual. As with...
For the catalase test a clean microscope slide was placed in a petri dish, fresh bacterial cells were smeared on the center of the slide, then a drop of 3% hydrogen peroxide was administered onto the bacterial cells, and formation of bubbles was observed. MSA test was conducted by obtaining a petri dish containing MSA medium and .5 ml tube. A sample of the bacteria along with 200 µl of sterile water are mixed in the tube, 5-10 sterile beads and 50 µl of the newly made bacterial solution are added to the MSA petri dish and the solution is spread out using the beads. The petri dish is then incubated for 2 days at 37°C, and evaluated for
Lactic Acid Bacteria (LAB) is a group of bacteria that is characterised by the production of lactic acid during the fermentation process of carbohydrates. They are further characterised as anaerobic, Gram positive bacteria that are also catalase negative and non-motile. In terms of morphology they are either rod-shaped (bacilli) or spherical (cocci). In addition, they all ferment carbohydrates and hydrolyse arginine. This group consists of six genera of bacteria – Enterococcus, Lactococcus, Streptococcus,
Bacterial cells, like plant cells, are surrounded by a cell wall. However, bacterial cell walls are made up of polysaccharide chains linked to amino acids, while plant cell walls are made up of cellulose, which contains no amino acids. Many bacteria secrete a slimy capsule around the outside of the cell wall. The capsule provides additional protection for the cell. Many of the bacteria that cause diseases in animals are surrounded by a capsule. The capsule prevents the white blood cells and antibodies from destroying the invading bacterium. Inside the capsule and the cell wall is the cell membrane. In aerobic bacteria, the reactions of cellular respiration take place on fingerlike infoldings of the cell membrane. Ribosomes are scattered throughout the cytoplasm, and the DNA is generally found in the center of the cell. Many bacilli and spirilla have flagella, which are used for locomotion in water. A few types of bacteria that lack flagella move by gliding on a surface. However, the mechanism of this gliding motion is unknown. Most bacteria are aerobic, they require free oxygen to carry on cellular respiration. Some bacteria, called facultatibe anaerobes can live in either the presence or absence of free oxygen. They obtain energy either by aerobic respiration when oxygen is present or by fermentation when oxygen is absent. Still other bacteria cannot live in the presence of oxygen. These are called obligate anaerobes. Such bacteria obtain energy only fermentation. Through fermentation, different groups of bacteria produce a wide variety of organic compounds. Besides ethyl alcohol and lactic acid, bacterial fermentation can produce acetic acid, acetone, butyl alcohol, glycol, butyric acid, propionic acid, and methane, the main component of natural gas. Most bacteria are heterotrophic bacteria are either saprophytes or parasites. Saprophytes feed on the remains of dead plants and animals, and ordinarily do not cause disease. They release digestive enzymes onto the organic matter. The enzymes breakdown the large food molecules into smaller molecules, which are absorbed by the bacterial cells. Parasites live on or in living organisms, and may cause disease. A few types of bacteria are Autotrophic, they can synthesize the organic nutrients they require from inorganic substances. Autotrophic bacteria are either photosynthetic or Chemosynthetic. The photosynthetic bacteria contain chlorophyll that are different from the plant chlorophyll. In bacterial photosynthesis, hydrogen is obtained by the splitting of compounds other than water.