Pseudomonas Aeruginosa In Blood Agar

Pseudomonas aeruginosa is a versatile gram negative bacterium that grows in soil, marshes, and coastal marine habitats, as well as on plant and animal tissues. People with cystic fibrosis, burn victims, individuals with cancer, and persons infected with HIV are particularly at risk of disease resulting from Pseudomonas aeruginosa.

In the LB (pGLO negative), it is expected to not see any colonies growing. As a result of this experiment, it shown any growth colonies but this only had shown a large number of white, circular colonies that were found across the surface of the agar. In the LB/Amp (pGLO negative), it is expected to see any growth colonies. In the experiment, the resulted was no growth colonies because this has Ampicillin and no pGLO. Now, the LB/Amp (pGLO positive), it is expected to have growth colonies in the agar plate. As a result, it was shown growth colonies in the agar plate because has positive pGLO regardless having Ampicillin. In the other hand, both positive pGLO have the same components but in one plate was added Arabinose. The LB/Amp/Ara (pGLO

This experiment was conducted to find the genus and species of an unknown bacteria prescribed by the lab teacher, which was unknown bacteria GA3 in my case. Identification of unknown bacteria techniques are used on an every day basis to figure out what type of bacteria it is and to find the best method of how to treat a patient with this bacteria (1). All five “I’s” of Microbiology were used in the testing for the unknown culture. Inoculation was used several times to put the unknown culture into agar plates or into biochemical test tubes. After Inoculation of these tubes or plates, they always were placed into the incubator for further growth and development. Isolation was used to make sure we got the correct bacteria we were testing for. After each further isolation, we gram stained the culture and inspected the culture under a microscope to further help in the identification process of the unknown bacteria. Multiple tests were done on the unknown culture to make sure we were confident in what kind of bacteria the unknown was.

There were two tubes used in this process: the tube that contained the primary culture and the tube that contained the nutrient agar where the unknown bacteria would grow. First, the inoculating loop was flamed. After removing the caps of both the test tubes, they were flamed to prevent contamination of the unknown bacteria. The inoculating loop was cooled for a few seconds and was then placed into the test tube containing the bacteria. The inoculating loop with the bacteria was placed into the nutrient agar test tube for cultivation. Before the test tubes were capped, they were flamed once again. Also, isolation of the unknown bacteria had to completed. Nutrient agar was placed in the petri dish, and was left to gel for a few minutes. After the agar gelled, the inoculating loop was used to acquire bacteria and streak the unknown onto the plate for

The purpose of this lab was to isolate and identify the microbes from the randomly assigned mixed culture in test tube number # 10. In order to achieve identification of the microbes the mixed culture will be inoculated on various media, and subjected to biochemical tests in order to differentiate and identify the microbes. These experiment were carried out over the course of five days in a microbiology lab. Isolation of two microbes for a pure culture of each was achieved by the streak plate method, and gram staining identified a gram-positive (+) microbe and a gram-negative (-) microbe. A blood agar plate was inoculated with the gram (+) microbe to reveal it was beta hemolysis.

In this experiment, pH indicators, colour indicators, metallic ions, and Kovac’s reagent all aide in the differentiation of different bacteria under different conditions. Proteus vulgaris for example is a rapid production of urease and this is shown through the phenol red indicator, turning from yellow to pink. This experiment is usually done to distinguish from Proteus and other bacteria, which helps to isolate these bacteria since they are infectious (O'hara, et al., 2000).

pneumoniae) leaving me with Sa, Se, Ef, P.pyo, and Ml. Following my dichotomous key, the next test I decided to perform was a catalase test in order to determine if the enzyme catalase is produced by the bacteria. The sample of the bacteria I took did bubble, this indicates that the bacteria does produce the catalase enzyme as well as requiring oxygen to respire. Based on the results of the catalase test there were three possible microbes I could have Ml, Sa, and Se. For my next test I decided to use an MSA plate to determine if my unknown microbe could or could not grow in the presence of the mannitol salt, along with testing to see if it would or would not be able to ferment the mannitol salt. After allowing the bacteria to grow, I noted that the plate remained the same pinkish color it originally was however, I did several white colonies growing on the plate. The pink color that remained shows that the organism is unable to ferment the mannitol salt thus the acidic byproduct that turns the medium yellow will not be produced. Therefore the organism cannot be Sa since that

Next the tubes were placed in an ice bath, while obtaining a sterile loop to swipe a single colony of E.coli to put into the tube. After gently swiping a colony onto the loop, it was then spun in the +pGLO tube to get it to come off, returned to the ice bath. Next using a different sterile loop, it was swooped it in a container labeled pGLO plasmid DNA and again spun it ONLY into the tube with the solution labeled +pGLO to get it to come off. After about 10 minutes on ice the tubes were then placed into a 42ºC water bath for 50 seconds exactly, and immediately after placed them back into the ice bath. Finally, after 2 more minutes in the ice bath the tubes were separated into 4 containers. 250 ul of +pGLO solution was added to the containers containing +pGLO, LB broth, with ampicillin and +pGLO, LB broth, ampicillin, with arabinose. Also 250 ul of the –pGLO solution was added to the 2 containers containing LB broth, and ampicillin, with LB broth. Using a sterile loop for each plate the solutions were spread out gently and thoroughly on to the containers with agar. After the containers were incubated in 37ºC for at least 24 hours, the results were observed and disposed of (Weedman,

A total of 13 tests were performed on the unknown bacteria culture #1. Two tests were inconclusive and could not be used. The first test was simply the phenylethyl alcohol agar (PAA) to check for contamination because of its selectivity towards gram-positive bacteria. The alcohol in the agar penetrates the thin peptidoglycan wall of gram-negative bacteria, which results in a slowed and stopped growth. The mannitol salts agar (MSA) is a differential test due to the mannitol sugar. It also has high amounts of salt (7.5% NaCl). If the organism can ferment the sugar, and produce acid as a result, the phenol red pH indicator dye will turn yellow due to the acidic environment. If no change occurred, then it is a negative result meaning that the bacteria

After the multitude of tests performed, it was determined that the bacterial unknown was Staphylococcus aureus. The gram stain slide was positive. The morphology and arrangement was grape like cocci clusters. On the glucose fermentation test the bacterial unknown tested positive for acid and negative for gas. The oxidase test was negative. The bacterial unknown tested positive for catalase. In the litmus milk medium a K or alkaline reaction was observed. It tested negative for urea hydrolysis. The bacterial unknown tested positive for nitrite. On the Kigler 's iron agar media it tested negative for gas, positive for glucose, positive for lactose, and negative for hydrogen sulfide. On the SIM medium media it tested negative for indole, negative for hydrogen sulfide, and negative for motility. The bacterial unknown tested negative for methyl red. It tested negative for Voges-Proskauer. It tested negative for citrate. It tested

The first day an unknown sample was assigned to each group of students. The first test applied was a gram stain to test for gram positive or gram-negative bacteria. The morphology of the two types of bacteria was viewed under the microscope and recorded. Then the sample was put on agar plates using the quadrant streak method for isolation. There were three agar plates; one was incubated at room temperature, the second at 30 degrees Celsius, and the third at 37 degrees Celsius. By placing each plate at a different temperature optimal growth temperature can be predicted for both species of bacteria.

Escherichia is a genus of aerobic gram-negative rod-shaped bacteria of the family Enterobacteriaceae that form acid and gas on many carbohydrates, such as dextrose and lactose, but not acetone, which include occasional pathogenic forms, including some strains of E. coli which are normally present in the human intestine as well as other forms which typically occur in soil and water (Webster). Escherichia coli is a gram-negative bacilli that rarely varies in shape and size and when stained often resemble safety pins because the ends of some bacilli stain more densely than does the middle; which is a characteristic called bipolar staining which is common in enteric gram-negative bacilli (ASM). Gram negative cells have a thin cell wall layer and will stain red to pink. The staining process is the same as Gram positive, requiring four steps: applying a primary stain, adding a mordant, then rapid decolorization and completing with a counter stain. Applying the alcohol for decolorization dissolves the outer membrane and leaves small holes in the thin peptidoglycan layer through which the crystal violet-iodine diffuse. The gram-negative bacteria is colorless after the decolorization; therefore adding safranin

The bacteria I was working with occupied a very small amount of space on the agar. The tiny spot, in which I called my bacteria, appeared to be an off white or creamy color and had much of a sticky looking texture to it, although I did not touch it. It was also very glossy and the colony was in a circular shape but only took up as much space as a small tick would. After going through all of the fun steps of gram staining, with the critical steps of inhibiting crystal violet, Potassium Iodide, Alcohol, and Safranin, the bacteria on my slide turned out to be much of a faded color of pink. Thereafter, I was then told that I had messed up terribly by pouring an excessive amount of alcohol onto my slide and it was then realized it was meant to be a dark purple color. With it being the

A great deal of cultural tests and physiological procedures were performed on the unknown before the conclusion of P. aeruginosa was made. The first test preformed was Gram’s staining which involves the use of crystal violet, Gram’s Iodine, Ethyl Alcohol, and Safranin. Gram staining is known as a differentiating stain, since it can determine the presence or absence of peptidoglycan in the cell wall. The unknown bacteria were Gram stain negative. After that the motility of the cell was tested using a hanging drop slide. The slide was examined, but the results were inconclusive due to the inability to make a clear distinction between motility and Brownian motion. This test was not

A fresh pipette was used to transfer 0.5ml of broth culture of E. coli, to be inoculated, into a tube of molten agar previously boiled to drive off oxygen. The tube was then rolled to distribute the bacteria and allowed time for the agar to harden. The tube was incubated at 37oC for 48 hours. The procedure was repeated for broth cultures of the bacterium Clostridium sporogenes and B. Subtilis.