Microbiology Unknown

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 purpose of this lab was to identify an unknown bacteria culture using differential tests. The identification of the unknown culture was accomplished by identifying the bacteria based on its specific metabolic characteristics and morphology. It is suggested that culture 11 is a sample of Enterobacter aerogenes.

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

In a laboratory setting, it often becomes necessary to identify an unknown organism. In this experiment, researchers classified an unidentified bacterium based on its physical structure, colony morphology, optimal conditions and metabolic properties. A Gram stain using crystal violet, iodine, and safranin and a simple stain using methylene blue characterized the organism’s cell wall. Cultural behavior was classified by inoculating the organism onto nutrient agar and incubating it at 37° C for 48 hours, and observing its behavior, as well as using SIM medium to test for motility. Optimal growth temperature was

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

|EMB Agar | |Distinguishes bacteria that ferment |Dark blue colonies with|E. coli and P. |

The two remaining sp2 hybrid orbitals on oxygen are used to hold oxygen's lone pairs(bruice). O-ethylsaccharin is then less stable because the Ethyl group is attached to the Oxygen that used to be a carbonyl group, giving the Carbon an sp3 configuration (joined to two other carbons, the Oxygen with the Ethyl group and a Hydrogen). This put strain on the ring, and therefore is less stable.”(Richard y.a.). Upon mixing the reactants, sodium saccharin slightly dissolved producing a clear colorless liquid. When placed at 80°C hot bath, the solution completely dissolved and turned yellowish-green. Iodoethane is a clear and colorless liquid, but when exposed to light and air the Iodide dissociates from ethane and gives off a yellow color as a sign of decomposition. The solution was covered to prevent this from happening. But, as iodide dissociates from CH3CH2 that then gave off its yellowish color which shows SN2 reaction taking place. SN2 reaction happened fast The limiting Reagent is Iodoethane , as the alkylating agent; it was not used in excess and dictated how far the reaction went. The Na+ binds with I-(noted disappearance of the yellow color, as I- binds with Na+) then the ethane group bonds with either the Oxygen of saccharin or the Nitrogen of saccharin. The final product after vacuum filtration had some unreacted material, indicated by some yellow green solid formation.

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

The experiment began by mixing the initial 1.775g isopentyl alcohol with 2.3 mL acetic acid and about 5 drops sulfuric acid. This reaction mixture was then heated under reflux for an hour after boiling of the reaction mixture began.

The filter paper was then observed to see if it changed blue or not, in order to see if the bacteria produced cytochrome c oxidase. The final test used in the experiment was an API test. To begin the API test, a solution with bacteria and 5 mL of sterile saline, had to be made with a turbidity the same as the McFarland No. 3 (BaSO4) standard. This was done by adding loopfuls of bacteria to the saline solution, mixing the solution on the vortex, and then comparing the turbidity to the McFarland No. 3 standard, until the tubes were both at the same cloudiness. This created solution was then used in the API test by adding specified amounts to each of the microtubes on the API strip. For each of the microtubes whose names were not underlined or boxed, the tubes were filled to where the microtubes met the capsule. In the microtubes whose names were underlined, the microtubes were slightly underfilled, and then the capsule was filled with mineral oil in order to create and anaerobic environment. The last of the microbes were the ones whose names were boxed. In each of these the microtube and the capsule were filled all the way up with the bacteria. The API test strip was then placed in the 37°C incubator for 20 hours. After this time, observations were made about each of the different microtubes based on a given summary of results chart for the API test. A select number of microtubes had

In the Voges-Proskauer test, I inoculated the tube with bacteria from my TSA plate and incubated the tube at 37 degrees Celsius for 3 days. After three days I placed some Barrits Reagent A and Barrits Reagent B in the test tube. The color change of red or pink indicates a positive reaction for acetoin which tells you that the organism is a butanediol fermentor.

As the flowchart shows, a series of tests were conducted to identify the unknown bacterium #65. Microscopic observation of the gram stain indicated a gram-positive coccus bacterium. S. epidermidis was used as the gram-positive control while E. coli was used as the gram-negative control. This observation led to the elimination of all gram negative and rod-shaped genera: Enterobacter, Citrobacter, Klebsiella, Escherichia, Pseudomonas, Serratia, Alcaligenes, Neisseria, Proteus, Salmonella, Shigella, Erwinia, Veillonella, Flavobacterium, Bacillus, Arthrobacter, Lactobacillus, Listeria and Kurthia (2). By performing the catalase test, it was determined that the bacterium was catalase negative and it did not produce bubbles. M. luteus and E. faecalis were used as positive and negative controls, respectively.

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

In part II of the lab six small glass tubes were obtained in a test tube rack. Ten drops of distilled water were then added to test tube 1, five drops to tubes 2-4, and no drops in tubes 5 and 6. Five drops of 0.1M HCl were added to test tube 5 and five drops of 0.1M NaOH to test tube 6. Five drops of enzyme were then added to all tubes except tube 1. Tube 3 was then placed in the ice bucket and tube 4 was placed in the hot bucket at 80-900C for five minutes, the remaining tubes were left in the test tube rack. After the five minutes five drops of 1% starch was added to every tube and left to sit for ten minutes. After ten minutes five drops of DNSA were then added to all the tubes. All the tubes were then taken and placed in the

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.