Starch
Only the Gram positive organisms were put through the starch test, as testing the Gram negative organisms would not have helped with identifying the unknown organism. Proper inoculation techniques were used to transfer the Gram positive organisms onto the starch plate in a straight line, and then the plate was incubated. The following week, after incubation, the plate was flooded with Gram’s iodine and was left to sit for ten minutes. Gram’s iodine indicated the presence of starch by forming a starch iodine complex that turns blue-brown or black. After pouring the excess iodine off, the plate was set on a white paper to analyze. A positive test with amylase present would present itself with a halo of clearing around the cell
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Citrate-negative cultures would also show no growth in the medium.
Urease
The urease test was not available, so students asked the lab TA for urease test results for their respective organisms.
Results
Gram Stain After Gram staining, organism A was classified as a Gram positive organism. On the other hand, organism B was classified as a Gram negative organism. After this, the two organisms were separated into their own flow charts. These results allowed for the strategic planning of tests of the Gram positive and Gram negative organisms to further identify them.
Fermentation
Organism A was tested positive for both glucose and lactose. Organism B was determined to be positive for glucose, but was tested negative for sucrose. The sucrose test significantly narrowed down the possible organisms for organism B. However, all the fermentation test results proved to be useful in eliminating possible organisms while confirming the correct identification of the two organisms.
Starch
Organism A was tested to be negative for the starch. This information helped in eliminating some possible options for the identification of the unknown.
Casein
Organism A tested to be negative for casein, which solely eliminated Micrococcus luteus as an option. Organism B tested to be positive, which eliminated some possible options for the unknown organism.
Catalase
Organism A was tested to be positive for the catalase,
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
This experiment was centered on metabolic and biochemical testing procedures. The rationale of performing these tests was to distinguish six different microbes from one another and to compare how their metabolic and biochemical processes differ from species to species to determine the unknown sample.
The purpose of this lab was to test different substances using various procedures to see what biomolecules were present and ultimately find out what restaurant Anna Lyza had eaten at before she died. For the first control test, we used vegetable oil to test for lipids. So, if the solution does not contain lipids, it does not become translucent when placed onto a paper bag square and held up to a light. So, it is a negative result. However, in the presence of lipids, the solution will become translucent when placed onto a paper bag square and held up to a light. Therefore in this case, the result is positive. On the other hand, we used albumin egg to test for proteins in another control test. If the solution does not contain proteins, it will not experience any color change and so it is a negative result. When there are proteins existing in the solution, it will turn bluish/purplish and for this reason it is a positive result. Furthermore in the third control test, we used dextrose to test for simple carbohydrates such as glucose. If the solution does not contain simple carbohydrates, it will not undergo any color change and will remain a blue color. So, it is a negative result in this circumstance. If there are simple carbohydrates present in the solution, the solution will turn reddish and so the result is positive. For the last control test, we used starch solution to test
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
11. An important tool available in the Virtual Unknown program is the Identification Matrix. From the portion of the identification matrix shown in the Identifying Bacteria tutorial, identify at least one bacterium that has a positive result to the arabinose fermentation test. (1 pt)
I began by running the starch test, which tests for the presence of starch hydrolyzing enzymes. After doing a one-line inoculation of the organism, the plate had to be incubated. Once I received an appropriate amount of growth I added the reagent iodine. The iodine turned the plate purple, formed no clear zone, and lifted the organism off of the plate, which revealed that the starch was not degraded and the enzyme was not present. The organism being lifted off the plate is unique to the bacteria Corynebacterium xerosis indicating that it was my gram positive rod. For reassurance, I ran the Phenol Red Glucose test, which tests if the organism contains various enzymes that determine if the bacteria can ferment glucose. After incubation, the broth turned orange, but this did not provide a clear positive or negative result so I ran the Nitrate Broth Reduction test. The Nitrate Broth Reduction test detects if the organism utilizes nitrate. After incubation for forty-eight hours I added Nitrate A and Nitrate B indicators. However, there was no color change indicating that the test was inconclusive. Since the test was inconclusive, I proceeded to the following step, which included adding a small amount of zinc to the broth, and this turned the broth a red color. The red color indicated that
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 I have been learned so far in the microbiology laboratory class for the identification of an unknown bacterium.
Many tests were completed on the unknown such as gram staining and inspection under microscopes to find whether the bacterium is gram positive or gram negative. Chemical resistance tests were also performed to see if certain chemicals affected the unknown growth or if it didn’t affect the bacteria at all. Each biochemical test
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
This experiment was given to us to utilized previous knowledge learned throughout the semester to identify a gram negative unknown bacterium. We had to first learn the difference between a gram negative and a gram positive organism. We started off with doing gram stains to determine whether it was positive or negative. Based on the gram stains, a gram positive stains purple and a gram negative stains pink. A gram positive stains purple because the cell walls is made of a thick peptidoglycan layer and doesn’t
Two smears of the unknown bacterium #5 were inoculated while the second smear was used for a back up. The unknown bacterium dried for at least forty minutes. After the smears dried, the slides were heat fixed two times to ensure the stability of the organism. The slide was placed on top of the staining rack then over the small sink.
Gram staining is a technique used to determine if the bacteria is Gram positive or Gram- negative. Gram staining procedure uses crystal violet stain, iodine moderator, alcohol decolorizer and safarin counter stain. In Gram- negative bacteria the primary stain will be washed out with the decolorizer and it will be stained with the counterstain. Whereas in Gram-positive bacteria the primary stain will not leave the cell wall. This difference comes from difference in the structure of the cell wall that retains the stain.
enzymes that will be used during this lab to test the ability of amylase to break down starch ,a
The bases of this experiment was to discover the identify of the unknown from three possible specimens: Klebsiella pneumonia, Escherichia coli, and Enterobacter aerogenes. Utilizaing the T streak technique, the bacteria was isolated into pure colonies for further study. The Gram Stain method was used to identity the morhphology of the bacteria such as the shape and whether the bacteria was Gram positive or Gram negative. Biochemical test were also used to help identify the unknown bacteria. The biochemical test used was the Triple Sugar Iron Agar, Sulfur Indole Motility test, Methyl Red test, Voges-Proskauer test, Citrate test, Urease test, and the Gelatin test. After observing the morphology of the bacteria using the Gram Stain method and utilizing all the possible biochemical test, the bacteria was identified to be Enterobacter aerogenes.
In class, we were given the task of identifying an unknown bacterium broth culture. After receiving number 69, I went through several tests to figure out what bacterium I received. First, I created a slide from my broth by putting a small amount of the unknown broth on to a clean slide and letting it dry for ten minutes. After this, I stained the slide by applying four reagents in order; crystal violet, grams iodine, decolorizer and safranin. From the stained slide, I discovered that this bacterium was gram-negative, which would determine the next couple of tests I would do to identify my unknown bacterium. I began by streaking for confluent growth from my broth culture onto a TSA plate. From the TSA plate, I aseptically transferred a loop