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Gram staining lab report
Gram staining lab report
Gram staining lab report
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Unknown Organism Assignment
Lacey Fagan
Microbiology: Mrs. Rossman
I was given unknown organism #14, in order to find out what organism I had, I had to perform several different biochemical tests to identify it. Starting with the Gram stain test, which is performed to differentiate Gram-positive and Gram-negative cells. After staining, when observed through the microscope Gram-positive cells are a purple color with thick peptidoglycan cell walls. Gram-negative cells are a pinkish/red color with thinner cell walls. (handout G. s.) My organism was observed to be pinkish rod shaped meaning it is Gram-negative bacteria. The Phenol Red Carbohydrate Fermentation Broth test. This test is to determine the ability of microorganisms to ferment in a specific carbohydrate. If fermentation occurs, the pH level will change and your broth will
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If the organism can use the Citrate it will produce alkaline by products that will raise the pH level of the medium and growth will occur, causing a positive result. If Citrate cannot be used there will be no growth and no change in color due to no pH level changes causing a negative result. My organism showed signs of growth and color change from green to blue, meaning a positive result for my organism. ( handout Citrate) The Amylase test is to determine if the bacteria produces amylase which cleaves the starch into di- and monosaccharides. Iodine turns into a blue/black color when in the presence of starch, after using iodine if the blue/black color is absent then the starch has been used usually making a halo around the inoculum, resulting in a positive result. If it stays blue/black then the starch is still present meaning the organism cannot produce amylase causing a negative result. My color stayed blue/black and there was no evidence of a halo, meaning my organism is negative for producing amylase. (handout,
Then, an amount of KI (solid) about a size that would fit on a match head was dissolved in 0.05 of Potassium Iodate solution and about 1 mL of water and 1 mL of 1 M HCl were added, which exhibited a weak positive test for IO_3^- (aq). After the weak positive test, an amount of KI (solid) about a size that would fit on a match head was dissolved in about 1 mL of water and 1 mL of 1 M HCl, which exhibited a negative
I identified the genus and species of an unknown bacterial culture, #16, and I applied the following knowledge of morphologic, cultural and metabolic characteristics of the unknown microorganism according to the laboratory manual as well as my class notes and power point print outs. I was given an incubated agar slant labeled #16 and a rack of different tests to either examine or perform myself; the tests are as follows: Gram Stain; Nutrient Gelatin Test; Carbohydrate Fermentation; Dextrose, Lactose and Sucrose; IMVIC tests; Citrate, Indole, Mythel-Red and Vogues Proskauer test; as well as a Urease and TSI Test. Materials and Methods/Results Upon receiving the Microorganism (M.O.) #16, I prepared a slide by cleaning and drying it. Then, using a bottle of water I placed a sterile drop of water on the slide and used an inoculating loop, flame sterilized, I took a small sample of the unknown growth in my agar slant and smeared it onto the slide in a dime sized circle and then heat fixed it for ten minutes.
One bacterium was gram negative. It underwent four different tests. These tests were the EMB test (Eosin Mehylene Blue), the Sulfur Indole Motility (SIM) test, the Urease test, and the Simmon’s Citrate Utilization test. The EMB test checks for a bacteria’s ability to ferment lactose. This test is accomplished by placing the bacteria on Eosin Methylene Blue agar. The agar is selective for gram negative bacteria and those bacteria that can ferment lactose will have colored growth, usually a metallic green sheen.
In this lab project, the microbiology students were given 2 unknown bacteria in a mixed broth each broth being numbered. The goal of this project is to determine the species of bacteria in the broth. They had to separate and isolate the bacteria from the mixed broth and ran numerous tests to identify the unknown bacteria. The significance of identifying an unknown bacteria is in a clinical setting. Determining the exact bacteria in order to prescribe the right treatment for the patient. This project is significant for a microbiology students because it gives necessary skills to them for future careers relating to clinical and research work.
In the last decade, the number of prescriptions for antibiotics has increases. Even though, antibiotics are helpful, an excess amount of antibiotics can be dangerous. Quite often antibiotics are wrongly prescribed to cure viruses when they are meant to target bacteria. Antibiotics are a type of medicine that is prone to kill microorganisms, or bacteria. By examining the PBS documentary Hunting the Nightmare Bacteria and the article “U.S. government taps GlaxoSmithKline for New Antibiotics” by Ben Hirschler as well as a few other articles can help depict the problem that is of doctors prescribing antibiotics wrongly or excessively, which can led to becoming harmful to the body.
Bacteria play a large role in our health, the environment, and most aspects of life. They can be used in beneficial ways, such as decomposing wastes, enhancing fertilizer for crops, and breaking down of substances that our bodies cannot. However, many bacteria can also be very harmful by causing disease. Understanding how to identify bacteria has numerous applications and is incredibly important for anyone planning to enter the medical field or begin a career in research. Having the background knowledge of identifying an unknown bacteria may one day aid healthcare professionals diagnose their patient with a particular bacterial infection or help researchers determine various clinical, agricultural, and numerous other uses for bacteria.
The independent variable for this experiment is the enzyme concentration, and the range chosen is from 1% to 5% with the measurements of 1, 2, 4, and 5%. The dependant variable to be measured is the absorbance of the absorbance of the solution within a colorimeter, Equipments: Iodine solution: used to test for present of starch - Amylase solution - 1% starch solution - 1 pipette - 3 syringes - 8 test tubes – Stop clock - Water bath at 37oc - Distilled water- colorimeter Method: = == ==
The scientific techniques that were mainly used in the conduction of our investigation were the identification of bacteria through the use of a microscope, putting together a Simple Distillation Kit, treating Petri Dishes with Agar on them with the boiled, distilled or radiated water and calculating the coverage of bacteria on the amount of area treated on the Petri Dish. All of these scientific techniques were a crucial part of our investigation. Without all of these techniques we wouldn’t have been able to conduct such a successful experiment.
Methyl Red: One can identify an organism’s ability to conduct the mixed acid fermentation of glucose in a positive test. The fermentation of glucose will lower the pH to less than 5 by overcoming the phosphate buffer that is in the medium6. The lowered pH will allow the indicator, methyl red, to turn the medium red.
There were five test solutions used in this experiment, water being the control, which were mixed with a yeast solution to cause fermentation. A 1ml pipetman was used to measure 1 ml of each of the test solutions and placed them in separated test tubes. The 1 ml pipetman was then used to take 1ml of the yeast solution, and placed 1ml of yeast into the five test tubes all containing 1 ml of the test solutions. A 1ml graduated pipette was placed separately in each of the test tubes and extracted 1ml of the solutions into it. Once the mixture was in the pipette, someone from the group placed a piece of parafilm securely on the open end of the pipette and upon completion removed the top part of the graduated pipette.
The Snyder test is to see if there is a susceptibility to any cavities. This test uses saliva and the Snyder ager. The saliva contains bacteria that ferment lactose to lactic acid, which can cause color change on the agar due to the pH indicator. As for the Tomato juice isolation test helps to be able to isolate bacteria that are from the normal flora. The use of Subouraud agar is a medium that can help detect fungi and yeast cells that are commonly within humans. The mouth contains low levels of yeast cells but having larger numbers can cause problems in the mouth. This experiment t will also be using saliva to test the microbial diversity in
However, in order to measure the rates of reaction, sodium thiosulphate and starch are added. Sodium thiosulphate is added to react with a certain amount of iodine as it is made. Without the thiosulphate, the solution would turn blue/black immediately, due to the iodine and starch. The thiosulphate ions allow the rate of reaction to be determined by delaying the reaction so that it is practical to measure the time it takes for the iodine to react with the thiosulphate. After the all the thiosulphate has reacted with the iodine, the free iodine displays a dark blue/black colour with the starch. If t is the time for the blue/black colour to appear, then 1/t is a measure of the initial rate.
Most of the results from the science experiments conducted did not occur like they were expected. Most of the results were false negative or false positive due to a human error within preparation of the bacteria slides. For example, the simple stain results for the Corynebacterium xerosis bacteria were inconclusive because they appeared to be very tiny circular dots instead of their usual rod shape. There is a possibility that it could have been the result of overheating the bacteria during the heat fixing process. This also could have been the result of leaving the crystal violet dye on for too long allowing the dye to dry up on the slide. The next example of an experiment’s results gone wrong is apparent in the Gram staining experiment. The
Knowledge is power when discussing the classes you have taken in college and how it affects your everyday life. Microbiology is one of those important classes where although for a majority of majors you do not have to take it and therefore will not learn the information discussed in it, other majors such as biology and nursing require you to take it. They will require you to take it because you will encounter some of the information being taught in it in the work field. As a nurse practitioner I have worked in the field with many patients who were diagnosed with Leprosy, Escherichia coli, Salmonella and other bacterial diseases. All of these diseases that I have encountered in patients were ones I was familiar with due to learning about them as a student in microbiology at CSUB. My children plan on