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
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
One bacterium was gram negative. It underwent four different tests. These tests were the EMB test (Eosin Mehylene Blue), the Sulfur Indole Motility (SIM) test, the Urease test, and the Simmon’s Citrate Utilization test. The EMB test checks for a bacteria’s ability to ferment lactose. This test is accomplished by placing the bacteria on Eosin Methylene Blue agar. The agar is selective for gram negative bacteria and those bacteria that can ferment lactose will have colored growth, usually a metallic green sheen.
The Gram positive bacteria has been nicknamed Posi. The Gram positive species’ morphology includes having an opaque opacity with a smooth margin. The moisture content of the Gram positive species is shiny and the pigmentation is gold. The Gram positive species grows at an optimal temperature of 37°C. The shape of the Gram positive species is a cocci, with an arrangement of grapelike clusters. The Gram positive species’ size ranges from .5-1.5 µm. Oxygen requirement of the Gram positive species is facultative, and has complete lysis of red blood cells. All results are summarized in Table
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.
I also inoculated a tryptic soy broth (TSB), a nutrient gelatin deep, a motility agar deep, a fluid thioglycollate medium (FTM) tube, and a TSA plate with my unknown culture. All of these inoculated media were incubated until the next class period (about 48 hours). Then when I came to class most of my inoculated tubes and my streak plate appeared to have growth. The next step I took was making a gram stain to determine the gram reaction and cellular morphology of my unknown. I used my working slant to do this, after careful examination of the gram stain, I learned that my unknown was a gram-positive bacterium. I then preceded by making a negative stain to see the size of the cells of my unknown bacteria. The cell shape was cocci and the cells occurred in clusters of tetrads. After discovering that my unknown bacteria was gram-positive cocci, I turned to page 207 of the lab manual to narrow down my options, there was only four out of the gram-positive list that were
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.
The purpose of this project was to identify unknown bacteria species from a mixed culture. The two unknown species were initially plated onto Tryptic Soy Agar (TSA), Eosin Methylene Blue (EMB), Mannitol Salt Agar (MSA), and blood agar plates to distinguish between the two different bacteria using colony size, color, shape, and growth characteristics. By identifying and inoculating the differing types of colonies, the two unknown bacteria were purified and able to be tested
The purpose of this investigation was to identify an unknown bacterium. “At any time there are millions of bacteria living around, on, or inside us” (The Plague). Bacterium can’t be identified by merely looking at it. Many bacteria have the similar appearances in growth. “In most cases, detection is based on the reaction of an enzyme with a certain substrate” (Sigma-Aldrich). Identification is usually based on the results of the bacterium’s cells metabolic capacities.
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
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.
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,
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.
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.
In order to know the morphology of bacteria, the gram staining procedure was conducted by using four different kinds of chemicals like Crystal Violet, as a primary stain; Gram’s Iodine, as the mordant for enhancing the crystal violet staining; 95% Ethanol, as the decolorizer for the Gram-negative cells and Safranin, as the counterstain, giving the Gram-negative cells its pink color.