The Effect Of Second Layer Matrix On Target Spots Unclassified By Maldi Tof

Proper collection of blood cultures are necessary and the most direct method of determining whether or not a patient is septic. The purpose of obtaining blood cultures is to identify and isolate the bacteria that are causing an illness and then determine the best course of treatment based on the sensitivity of the bacteria to particular antibiotics. One of the most frustrating problems plaguing hospitals is the increased rate at which blood culture results are being returned as contaminated specimens. These results can lead to a significant increase in cost to the hospital and patient as well as an increased length in hospital

Gold standard procedures should be implemented with the aim of providing timely and accurate results. (Schentag) The culture result should be accompanied by its clinical significance, selective reporting of susceptibility testing results in accordance with hospital antimicrobial therapy guidelines and suggested management. (Schentag) This encourages appropriate prescribing and minimises unnecessary use of antimicrobials. The clinical microbiology team should also provide reports to AMS regarding resistant organisms.

6. The disks in the 0.00% solution were transferred to an agar plate held next to the blue flame using the sterilized tweezers. Excess disinfectant was removed from the disks by wiping on the side of the well of the spotting tile. When the 5 disks were positioned (refer to Figure 1 below) the lid was replaced and sticky taped down. A label was added indicating the concentration of disinfectant.

Purpose: The purpose of this lab is to help you become a little familiar with some of the tests that can be typically performed in a clinical or research lab facility. These tests may help in determining a particular pathogen’s growth needs.

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 first step to identifying the unknown bacteria residing on the blood agar plate sent in from Khokana was to do a Gram stain on it. This is an important first step because it dictates further testing that will be necessary to arrive at a final conclusive result. Viewing the fixed and stained slide under the microscope revealed round chains of bacteria in a purple color signaling Gram-positive streptococci. A catalase test was performed with no bubbling present indicating a negative result. This further confirmed the shape and arrangement seen under microscopy. With this mind, the coagulase test was not done, as it would be of no use since that specifies for staphylococcus, specifically for Staphylococcus aureus. For streptococcus, an examination of hemolysis was necessary at this point. Shifting attention back to the original blood agar plate, gamma hemolysis was noted, thus narrowing the field down to two choices left. The unknown bacteria was either Streptococcus bovis or Entercoccus faecalis. This also means the Optochin and Bacitracin sensitivity tests would not be needed as those pinpoint alpha- and

Lastly, safranin was added to flood the slide and was left on for 1 minute and rinsed with distilled water. After adding all 4 reagents, the slide was blotted dry with a paper towel to remove any excess reagents or water. The slide was then placed on a microscope stage to look for the color of the bacteria. When ready for the highest power on the microscope, immersion oil was added to the slide and looked at under 100x power. A glucose test and citrate test were found to be the best two tests to help narrow down the bacteria.

Lab Day 1: After receiving my unknown bacteria, I streaked a TSA plate and incubated at 37°C for 48 hours. I then picked a single colony from the plate with my sterile loop and streaked a TSA slant and labeled it “Working Stock”. I did the same with another TSA slant and label the second one “Back-up Stock”. This would be the samples I used to complete the following procedures through the next four weeks to determine my unknown bacteria.

The importance of identification of a certain microorganisms can range between a life threatening diseases to a creation of certain antibiotic. Understanding the principals of living microbes and identifying my unknown bacteria through numerous biochemical and metabolism tests, with the outmost confidence, Proteus vulgaris had the precise qualifications. The point of this report is to further explore the identification of my unknown bacteria by revealing the results of the experiments and comparing them to the other six known bacteria: Micrococcus luteus, Staphylococcus aureus, Staphylococcus epidermidis, Alcaligenes faecalis, Escherichia coli, and Proteus vulgaris that were used in the lab, as well as comparing and

The purpose of this lab was to identify an unknown microorganism using lab techniques. The importance of identifying microorganisms is essential to the survival of humans, expansion of modern day medicine and improvement of quality of life. In 1884, Hans Christian Gram designed a differential staining technique to identify bacteria that would change the future of microbiology. He give rise to a staining process, known as the Gram stain to differentiate microorganisms into two groups between positive and negative gram staining microorganisms. The Gram stain is essential in a lab technique as it distinguishes the cells based on the physical properties of the individual cell walls, and is almost always the first test preformed to differentiate a microorganism. The identification of weather a microorganism is gram positive or negative can revel the bacteria’s virulence, cell wall structure, resistance to antibiotics, resistance to physical disruptions and so much more. In order to identify the unknown provided, unknown #27, the Gram stain was the first test preformed. After discovering that the unknown bacterium was indefinitely a gram positive microorganism, the vast possibilities were narrowed down. However, In order to more definitively identify the unknown, the next step was to preform biochemical tests. A biochemical test identifies metabolic

On a thin chromatography plate, five spots were placed ( as shown in table 2) and the plate was developed using chloroform/methanol. This was later visualized with dragendorff’s reagent under the UV light. All separated components were observed, identified and recorded.

In the 10-3 pasteurized sample, the plate exhibited 71,000 cells/mL. The results of the additional dilution samples contained too few colony forming units to count. However, in the 10-7 dilution, although the plate demonstrated 12 colonies, there should have been no colony forming units on this plate. The reasons for this could have been that this sample was contaminated from “double-dipping” the sample before dispensing it onto the plate or when using the pipette, it mistakenly was inserted in a higher concentration sample and then immediately to a lower concentration sample before it was dispensed onto the plate.

Materials and Methods The Materials involved in this lab include: saline solution, micro centrifuge tubes, micropipettes, a plastic cup, an ice bath, primer mix, nuclease free water, PCR enzyme mix, 1% Agarose gel, and loading gel. Day 1 of the experiment, we started by using a permanent marker to label a 1.5-mL tube and paper cup with our assigned number. Next, we gently rinsed your mouths out with water at the sink to remove food debris. Then, poured saline solution into our mouth (5-10ml), and vigorously rinsed our cheek pockets for 30 seconds, and expelled saline solution into the paper cup. Next, we swirled the cup gently to mix the cells that may have settled to the bottom, then use a micropipette with a fresh tip to transfer 1500

On the first day of this exercise, after using apsetic technique I inoculated a CNA .I moistened the swab with sterile water and before picking up a sample of bacteria, I taped the bottom of the broth against the palm of my hand. I dipped the swab into broth culture labeled #2 and smeared the entire surfaced of the CNA plate, I then turned the plate and swabbed the plate in a perpendicular direction to the first streaking creating a lawn. I used a T-soy plate to perform a streak isolation. Using apsetic technique and a loop, I obtained a sample of bacteria from the test tube #2, smeared the bacteria in a vertical smear, then streaked through the fist smear 10 times from the edge of the plate inwards, repeated the same procedure 4 times

The purpose of this experiment is to obtain isolation of individual species of particles from the mixed culture. This is completed through the isolation technique of streak plate. The objective of this experiment is to replicate the technique of streak plate but on a much larger scale. Because it is on a larger scale the particles are able to be visually observed as they are isolated using the streaking technique as the experiment is conducted. The benefits of the streaking technique is when a cultures has multiple species they are able to be more easily identified once they have been isolated. This experiment is much like the experiments completed on an agar plate but on one a much larger scale and where techniques