Petri Dish Lab

According to the series of test that my group ran for our unknown specimen, we had a match with the bacteria known as Alcaligenes Faecalis. This bacterium belongs to one of the major group of gram-negative bacteria (Phylum Proteobacteria). Alcaligenes Faecalis (Genus, species) is a rod shaped (bacillus), 0.5-1.2 x 1.0-3.0 µm, round with scalloped margin (colony configuration growth), motile (with one to nine peritrichous flagella), gram-negative, non-fermentative bacteria, obligate aerobic, having oxygen as the principal terminal electron acceptor in the electron transport chain (ETC). We consider we have a match with the species Alcaligenes Faecalis because of the following reasons: Fermentation tests performed (Durham sugars) were negative, which indicate that our bacteria use a different metabolic means for growth (non-fermentative gram-negative bacteria).

In the laboratory, identification of an unknown bacterium is often necessary. In the lab, a random sample consisting of three different bacteria was selected. The sample contained one gram-positive, one gram-negative paracolon, and one gram-negative coliform. The purpose of the experiment is to identify each of the three species that the mixture contained. After receiving an unknown mixture, the sample was streaked for isolation onto TSA, blood agar, and MacConkey plates.

After a gram stain was done unknown #257 was identified as a gram positive organism because when observed under the microscope the organism appeared purple with cocci in clusters. The organism was also catalase positive which means that it produced enzyme catalase and bubbled when hydrogen peroxide was added to it. Three test were conducted based on the result of the gram staining procedure. Blood agar with a Novobiocin disk was chosen as well as DNase (DNA) and Mannitol Salts (MSA) agar. The Blood agar is a bright red, opaque plate and the streaking or the inoculation technique was a modified streaking for isolation with a heavy quadrant one.

Biochemical tests are the tests used for the identification of bacterial species based on the differences in the biochemical activities of different bacteria. Bacterial physiology differs from one species to the other. These differences in carbohydrate metabolism, protein metabolism, fat metabolism, production of certain enzymes and ability to utilize a particular compound help them to be identified by the biochemical tests. Gram’s stain was originally devised by histologist Hans Christian Gram in 1884. Gram-positive bacteria stain purple, while Gram-negative bacteria stain pink when subjected to Gram staining.

Introduction Our world is composed of many bacteria’s’ that can either help or destroy us. Therefore, its’s imperative to learn and study them. The purpose of the lab was to put into action the methods that have been learned in the laboratory to determine our unknown bacteria. Bacteria’s can have different features, shapes, and or arrangements that help microbiologist determined their role in our life (whether they are good or bad for humans).

Joshua Miller 12/18/17 Fermentation Lab report Introduction The term fermentation refers to the chemical breakdown of a substance by bacteria, yeasts, or other microorganisms, typically involving effervescence and the giving off of heat (wikipedia). Sugars are converted to ethyl alcohol when fermentation happens. In this experiment we determined if yeast cells undergo fermentation when placed in a closed flask with no oxygen. Glucose and yeast are mixed together in a closed flask and allowed to incubate for about one hour.

The colour of each test tube was recorded and if proteins were present that was recorded for each test tube. Finally, the pH was recorded for each sample using pH

1 “substrate” and another “ enzyme.” Instead of using the distilled water, this time you are going to use different pH buffer in the enzyme test tube. In the substrate tube, add 7 mL of distilled water, 0.3 mL of hydrogen peroxide, and 0.2 mL of guaiacol for a total volume of 7.5 mL. For the enzyme tube, instead of distilled water add the pH solution (3) and 1.5 mL of peroxidase which equals a total volume of 7.5 mL. Use the dH2O syringe for our pH solution. To clean the syringe, flush it by drawing 6 mL of distilled water.

My bacterium turned out to be gram positive. When conducting these tests, I only had to do the coagulase test and the catalase test because when doing the catalase test, the reaction was that it had bubbled. If it did not bubble, or have a positive reaction, then I would not have had to do the coagulase test. Also, since my bacterium caused a positive catalase test, I only had to do the coagulase test and no other tests. This is because with staphylococcus organisms, these are the only tests

Being able to identify unknown microbes from systematic testing is what makes the field of microbiology so important, especially in infectious disease control. Using the testing procedure laid out by the microbiology field we are able to identify unknown bacteria present in our everyday lives, and along the way learn a lot about their characteristics that separate them from other types of bacteria. Being able to do this is vital in order for us to understand why microbes are present in certain places, how they are able to grow and what restricts their growth, that way they can be combatted if necessary. These techniques for determining unknowns are also important for isolating and testing infectious disease microbes in order to prevent spreading. Another important aspect of being able to identify unknown microbes is the

5 water bath were set up each to10 °C. (5 were used do the experiment faster) 5 cm3 of starch solution were added into the 5 test tubes that were labeled test tubes. Then 5 cm3 of amylase enzyme was added into the other 5 test tubes that were labeled. Put one of the starch solution test tube (preferably the one labeled 1) and one of the test tube containing amylase into the water bath (10 °C).

Introduction: Transforming a gene or genetic information from one organism into another with the hopes that if done successfully the organism with the new DNA will be given new traits is a method known as genetic transformation (Rafter). Genetic transformation is used quite frequently in today’s world, form medicine to agriculture.

Meanwhile, on our LB/amp plus DNA petri dish we had a lawn of bacteria and the color was almost clear. Our results for this petri dish mean that the bacteria was not killed by the ampicillin. Next, in the LB/amp minus DNA petri dish we observed very minimal amounts of growth. These results mean that we correctly followed the procedures because this petri dish was not supposed to have growth due to the ampicillin. Our last petri dish was minus DNA and lysogeny broth, in this petri dish we saw 7 colonies of bacteria and there were a light yellow color.

Methods Unknown microbial #398 went through several of tests in order to identify its characteristics when isolated from a urine sample of Doris, a 64- year old patient with a kidney infection. To identify unknown #398, must prepare a working and a reserve stock by the inoculation from a broth culture and by quadrant streaking method on a PEM and EMP plates. The following test procedures were incubated at 37°C for 48 hours for observation and identification for unknown #398. The identification of unknown #398 followed test procedures from Brown1.

After experiment on microscope under oil immersion, I learned that my Unknown is gram positive. Under the lens, the bacteria appears in purple color. Its morphology is cocci arranged in cluster. However, during decolorizing process, I put too much alcohol to the crystal violet-iodine complex making the color overly removed. That led to the result of my gram positive has slightly redish