Gen III Microplate Essay

In this experiment, an unknown bacterium was given to each individual student. The main purpose of this lab was to identify the given unknown bacteria going through a series of biochemical tests as one of the gram negative bacteria among six different Gram negative bacteria Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa and Salmonella typhimurium. At the very beginning, streaking method; T-streak technique was used to isolate the pure colonies. For the morphological identification of unknown bacteria, Gram Stain Method was done. Biochemical tests that were conducted for the experiment

The purpose of this study was to determine what an unknown bacteria was using several different microbiology lab techniques including an API test, an oxidase test, a gram stain, a hanging drop slide, and morphology identification. The unknown bacterium, which was contaminated with Serratia marcescens, was isolated by streaking the bacteria solution to single colonies. The isolated unknown white bacteria, had the appearance of circular form, convex elevation, entire margin, elongated cocci. The tests than showed that the bacteria was gram-negative, non-motile,

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.

The purpose of the bacterial unknown independent study experiment completed throughout the course of this lab was to determine the identity of an unknown bacterial species. The unknown bacteria sample was chosen from numerous samples provided by the instructor. The starting unknown sample, unknown #15 was a mixed bacterial culture and a broad approach taken to identify the sample. Various biochemical tests were completed to identify the bacterial species along with the use of databases such as Gideon and Bergey’s to compare the test results of known bacteria to the results of the unknown sample. Information was gathered from the other sources and databases and phenotypic testing completed and the results compared to the database results. Aseptic

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

The main objective was to identify an unknown organism by utilizing skills we learned in our labs this semester. The purpose was to attain the possible identity of the unknown organism by actually performing biochemical tests and staining techniques we learned in lab. After performing and analyzing the results, we were able to use Bergey’s Manual of Systemic Bacteriology as a guideline to narrow down the genus of our organism test by test.

The most common medium is blood agar. P. aeruginosa has very simple nutritional requirements. When grown on blood agar, it is known to form round colonies with a fluorescent greenish color under ultraviolet light due to the production of pyocyanine. It also gives off a sweet odor and shows a B-hemolysis. P. aeruginosa is known to produce acid, but no gas when in glucose. It is not an active fermenter of carbohydrates. The optimal growth is 37˚ C. It is oxidative and non fermentative. Although an aerobic atmosphere is necessary for optimal growth, it sometimes can be grown anaerobically if nitrates are present in the medium. It is easily differentiated from other bacteria when grown properly with no

Immediately do dilutions to 10-6 and plate out all dilutions. • The plate done at time 0minutes are the control plates. • Place the bacterial culture in a 65ᵒC water bath and begin timing with a stopwatch. • Dilutions to be done at 5,10,15,20 and 30 minutes withdraw 1 mL sample (don’t remove the sample from water bath) and make dilutions up to 10-6.

In the world of microbiology it is vitally important to be able to discern the identities of microorganisms. Not only is it important in a lab setting but as well as in healthcare in general. Properly identify what strain of bacteria a person has will aid in the proper medicine and dose given. Throughout the semester we have learned about different types of bacteria and certain test that can clearly identify them. The purpose of this lab report is to identify a Gram-positive or Gram-negative bacterium. Using all the knowledge of procedures and lab techniques identify the unknown and discuss all the tests you performed.

Read in your lab manual about the following agar mediums: Blood Agar (pg 168), EMB Agar (pg 170), Mannitol Salt Agar (MSA)(pg 172) ), MacConkey Agar (pg 174), and PEA Agar (pg 176) to answer the following:

A mixed culture of two unknown bacteria was provided by the instructor. The methods used for

You will need 11.5 grams of Nutrient Agar mix, a flask, a stir bar, a hot plate, a polystyrene partitian sterile petri plate, and 500 mls of distilled water. You now have to combine the agar mix and the 500 mls of distilled water and mix together. Put the agar into the 500 mls of distilled water and make sure the stir bar is in the flask. Put this mix on the hot plate and bring to a boil. Make sure the agar mix is dissolved before removing from the hot plate. After pouring into the plates, close the lids on them to prevent condensation. Also important is to use plates within one day of making agar. Make sure to label the plates properly(name, date, swabbing area). What is especially important during swabbing is to use sterile cotton Q-tips and to make sure they are not contaminated. To do this, you should not touch the tip of the swab or put it in contact with anything besides the area being tested. This general rule should also apply to the plates. To swab, after collecting the sample, gently roll the tip of the swab against the agar in a zig- zag type motion, making large patterns. This will help you to achieve better colony growth. Make sure to place the lid back on the plates. After swabbing, place petri plates in 37 degree celsius incubator and let stand for 4 days, then record data. When collecting data, make sure to use qualitative and quantitative

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.

Live Lactobacillus casei Shirota strain, cultured and tested in our laboratory, is added to the tank. The temperature of the tank is then reduced until the contents are at 37°C (body temperature). The solution is allowed to ferment in the tank for 6-9 days or until the numbers of Lactobacillus casei bacteria reach their ideal concentration.

The high accuracy tissue microarray instrument obtains tissue cores from predefined areas of tissue sample paraffin blocks and places them precisely on an empty recipient block. Tissue sections of 5 mm can be prepared from the tissue microarray blocks to generate tissue microarray slides for molecular and IHC applications. (Kittaneh et al.2013)