Lab Report

This experiment was centered on metabolic and biochemical testing procedures. The rationale of performing these tests was to distinguish six different microbes from one another and to compare how their metabolic and biochemical processes differ from species to species to determine the unknown sample.

This laboratory experiment’s objective was to take a pure culture and isolate it from a mixed culture. The other part of the objective was to ascertain what species of bacteria that the pure culture was. The hypothesis made stated that so long as lab protocol was followed, the unidentified culture would be positively recognized/identified. An isolated pure colony of the unknown culture was obtained using the quadrant streak plate method. Afterward, the culture was Gram stained, and the results showed that it was Gram positive. Motility tests were done on the unknown using a filter paper bridge on a petri dish that contained TTC with agar. The unknown was revealed to not be motile, which meant that it did not possess flagella. The last test done was to learn the metabolic capabilities of the unknown bacteria. There were tests done for citrate utilization, the mixed fermentation pathway, catalase presence, carbohydrate fermentation in mannitol, lactose and glucose, urease production and the butanediol fermentation pathway in order to better identify the unknown bacteria. The results from each of the metabolic tests in conjunction with the motility and Gram staining tests were ultimately compared to results from database containing many different kinds of results from various bacteria. The unknown from the mixed culture was identified as Staphylococcus

Fermentation is a metabolic pathway that produce ATP molecules under anaerobic conditions (only undergoes glycolysis), NAD+ is used directly in glycolysis to form ATP molecules, which is not as efficient as cellular respiration because only 2ATP molecules are formed during the glycolysis. One type of fermentation is alcohol fermentation, it produces pyruvate molecules made by glycolysis and the yeast will break it down to give off carbon dioxide, the reactant is glucose and the byproducts are ethanol and carbon dioxide. In this lab, the purpose is to measure whether the changes of

PH can affect the way fermentation occurs due to the irregularity of the acidity or alkalinity within the glucose solution. This is an enzyme-based reaction that is susceptible to pH. The aim of this experiment was to determine how pH affects the yeast fermentation rate by performing the experiment numerous times with a different pH of glucose solution which included pH 3, 5, 7, 9, 11. The hypothesis was ‘If the pH is lower than the neutral point then the fermentation reaction will occur faster?’ The experiment conducted was to measure the amount of C02 produced by the yeast going into fermentation, however varying the pH of glucose solution by using different pHs . To test this every 5 minutes the volume of gas in the test tube was observed and recorded until a period of 30 minutes had been. The end results

The mole is a convenient unit for analyzing chemical reactions. Avogadro’s number is equal to the mole. The mass of a mole of any compound or element is the mass in grams that corresponds to the molecular formula, also known as the atomic mass. In this experiment, you will observe the reaction of iron nails with a solution of copper (II) chloride and determine the number of moles involved in the reaction. You will determine the number of moles of copper produced in the reaction of iron and copper (II) chloride, determine the number of moles of iron used up in the reaction of iron and copper (II) chloride, determine the ratio of moles of iron to moles of copper, and determine the number of atoms and formula units involved in

11. An important tool available in the Virtual Unknown program is the Identification Matrix. From the portion of the identification matrix shown in the Identifying Bacteria tutorial, identify at least one bacterium that has a positive result to the arabinose fermentation test. (1 pt)

I identified Citrobacter freundii, the gram negative rod, by running a series of tests. I began with the Phenol Red Lactose tests, which tests if the organism contains various enzymes that determine if it can ferment lactose. The broth turned yellow after it was incubated, indicating that the lactose was fermented to acid, and there was also gas present in the Durham tube. Since the Phenol Red Lactose Test was positive, I then ran the Phenol Red Sucrose test, which tests if the bacteria contain different enzymes that determine if sucrose can be fermented. After incubation, the broth was yellow, indicating that sugar was fermented to acid, and there was also gas present in the Durham tube. Next, I ran the Sulfide Production, Indole Formation, Motility test, but I was only testing for Hydrogen Sulfide Production to differentiate between the organisms Citrobacter freundii and Enterobacter aerogenes. This test detects if the organisms can metabolize sulfur into hydrogen sulfide, which is revealed by the formation of ferrous sulfide that causes blackening around the growth. The test also reveals if the organism can break tryptophan into indole or migrate away from initial stab area. After incubation, the agar slant was completely black, indicating that the organism produces hydrogen sulfide and is motile proving that it was Citrobacter

For the cell to generate adenosine triphosphate (ATP), plenty of oxygen is needed. Fermentation is a unique way to generate energy ATP in the form of without using any oxygen. Fermentation consists of glycolysis and a process of NAD+ generation. Glycolysis oxidizes glucose to two pyruvate molecules. The two pyruvate molecules are converted into two carbon ethanol molecules, and two carbon dioxide molecules are released. The overall net yield of fermentation are two ATP molecules and two NADH

Cellular respiration is the series of metabolic process by which living cells produce energy through the oxidation of organic substances. Cellular respiration takes place in the mitochondria. Fermentation is the process by which complex organic compounds such as glucose, are broken down by the action of enzymes into simpler compounds without the use of oxygen. The significance of these pathways for organisms is to allow for an organism to be able to generate ATP. Some organism that undergo cellular respiration are bacteria and fungi. Some organism that undergo fermentation are yeast and muscle cells. In cellular respiration, glucose is oxidized and releases energy. In cellular respiration, glucose produces ATP and 3-carbon molecules of pyruvate. The pyruvate is then further broken down in the mitochondria where it becomes oxidized and releases CO2 (Upadhyaya 2014). In the fermentation process oxygen does not play a part. This process converts glucose into pyruvate and produces ATP. From there pyruvate breaks down into CO2 and acetaldehyde (Upadhyaya 2014) Monosaccharides are known as simple sugars and their main function is being the source of energy for organisms. Disaccharides are two monosaccharides joined by a covalent bond and their primary function is to provide food to monosaccharides. Some disaccharides

Bacteria groups or species can be differentiated by the fermentation patterns. The end-product of carbohydrate fermentation is an acid or acid with gas production and is dependent on the organism involved in the fermentation process. The carbohydrate fermentation tests detect if an organism is able to utilize glucose, lactose and sucrose. Phenol red is used as a pH indicator because it can indicate a change in pH when acid products are formed. Bacteria can utilize certain sugars resulting in an alkaline by-product which changes the color of the carbohydrate broth from red to yellow. Bubbles trapped within the Durham tube indicate the production of gas. The Phenol red carbohydrate fermentation tests determine that my organism E. coli can utilize glucose, lactose and sometimes sucrose but can only produce gas in glucose and lactose. (Phenol red carbohydrate fermentation lab

Furthermore, microbes are key component in food industry. Fermentation processes can be carried out by using the microbes like lactic acid, yeast, moulds and bacteria to make food products like beer, bread, yoghurt and cheese. For example, the bacterium Lactobacillus acidophilus is used to produce yogurt. Besides, bacterial acetic acid fermentation can produce vinegars. A microbe called Aspergillus sp. is used in the production of alcoholic beverages and also commonly used in the large-scale fermentation in the production of Japanese

With the results of the Starch Hydrolysis Test I preceded to the final test which is the Citrate Test. Using aseptic technique, I streaked the organism onto the surface of the Simmons’ citrate slant and let it incubate overnight at 37°C. The Citrate Test determines if the organism is able to

Organism A was tested positive for both glucose and lactose. Organism B was determined to be positive for glucose, but was tested negative for sucrose. The sucrose test significantly narrowed down the possible organisms for organism B. However, all the fermentation test results proved to be useful in eliminating possible organisms while confirming the correct identification of the two organisms.

Identification of the bacterial strains:The strains were subjected to gram staining,catalase and spore formation tests. (Harrigan and McCance, 1976).All Colonies were characterized in MRS and M 17 agars.The strains that gave gram positive and catalse negative results were set aside for further identification.(Sharpe, 1979).The growth of the bacteria at different temperatures of between Growth 10-45°C for 3-6 days , resistance to 60°C for 30 min (Sherman test), growth in the presence of 2 to 6 % NaCl and different pHs (4.5 and 6.5) were used to identify the strains of LAB. Arginine and asculin hydrolysis,citrate utilistaion, acetone productionformation of gas from glucose and production of dextran from sucrose were also determined. The starins were then tested for fermentation of L-arabinose, D-xylose, galactose, D-fructose, sorbitol, lactose, melibiose, saccharose, D-raffinose, melezitose, mannose and glucose.Bacterial growth in the different temperatures were confirmed by turbidity change in MRS or M17 after incubation(after 24,48 and 72 hrs).Microbial

The Voges-Proskauer test used the Methyl Red and Voges-Proskauer broth to determine if an organism produces acetoin after the degrading of glucose during a 2,3-butanediol fermentation (Leboffe, A Photographic Atlas for the Microbiology Laboratory). Only one bacteria,