Staphylococcus aureus was first identified by Sir Alexander Ogston in 1880, although Friedrich Julius Rosenbach is credited with giving the bacterium its current name in 1884. Staphylococcus aureus is commonly considered a natural inhabitant among the microfauna that is found on a human’s skin, inside the nostrils, and inside the lower reproductive tracts of healthy women (Kluytmans et al, 1997). Although S. aureus is commonly found on humans, it is known to be pathogenic, causing a range of illnesses, such as minor skin infections, pneumonia, toxic shock syndrome, and sepsis (National Institute Of Allergy And Infectious Diseases, 1999). Several other species of bacteria have had their names changed since they were first discovered, such as …show more content…
To establish whether the culture was pure, an isolation streak was performed using an agar plate, then both the TSB broth and the isolation streak were incubated at 35°C. First, a gram stain was performed to determine whether the bacteria were gram positive or gram negative, as well as to determine the shape of the bacteria. After the gram stain was performed, a series of biochemical tests were performed to help ascertain the identity of the unknown culture. Using an empty sterile test tube, a desiccation test was run to determine whether the bacteria formed endospores or not, by incubating it at 55°C for one week, followed by 24 hours at 35°C in a TSB broth. A catalase test was performed, by incubating the bacteria on an agar slant for 24 hours at 35°, to check for the presence of catalase. Afterwards, a SIM test was run to determine the motility of the bacteria, by incubating the bacteria for 24 hours at 35°C on an agar deep. A sample of bacteria were also streaked onto a tryptic soy agar (TSA) plate, and placed in a GasPak chamber for 48 hours to determine the bacteria’s oxygen requirements. A carbohydrate fermentation test was performed to determine whether the bacteria utilized glucose, lactose, or sucrose for fermentation, incubating …show more content…
The gram stain, when viewed under a microscope, determined that the bacteria were a gram positive coccus. The bacteria were not endospore formers, as determined by the negative results of the desiccation test, indicating that the bacteria had died. When in contact with hydrogen peroxide, the bacteria rapidly began producing bubbles, giving a positive result for the oxidase test. The results of the SIM test were negative for motility, as seen in the lack of growth throughout the tube. Due to the growth present inside the GasPak, the bacteria were revealed to be facultative anaerobes. The results for each carbohydrate fermentation came back positive, as all tubes had changed from red to yellow, indicating fermentation of each of the carbohydrates. When a colony was spread onto filter paper covered in oxidase reagent, the culture did not turn purple, indicating a negative result for the oxidase test. The ONPG test also yielded negative results, as the salt stayed transparent instead of turning yellow. The culture grown in the water bath showed the bacteria could grow at 45°C, and the cultures grown on the P-agar plate were larger than 5mm. The urease test resulted in a weak positive outcome on the slant, and a negative result in the bottom of the
An unknown bacterium was handed out by Dr. Honer. The appropriate tests were prepared and applied. The first procedure that was done was the gram stain. Under a microscope, if the gram stain is purple, the bacterium is gram positive, if the stain is red, it is gram negative. The next test was the fermentation tests for glucose, sucrose and
The purpose of this lab was to identify an unknown bacteria culture using differential tests. The identification of the unknown culture was accomplished by identifying the bacteria based on its specific metabolic characteristics and morphology. It is suggested that culture 11 is a sample of Enterobacter aerogenes.
Depending on what a bacteria can and cannot do, will help to correctly identify it. It is always important to start out with a purity check. To achieve that, you can inoculate an agar plate and incubate for 48 hours. Make sure you only see one type of colonies on the plate. Knowing the optimal temperature for the bacteria will also let a scientist know where to place the different types of medias he/she inoculates with the unknown bacteria. Knowing if you are working with a gram positive or gram negative bacterium, a scientist will need to perform a gram stain. This will also help to see the shape and arrangement of the bacteria. The size can be determined by doing a simple stain. The size of most bacteria ranges from .5- 10um. This specific bacteria that I was working with, was smaller than 2um. Most bacteria can grow with the presence of oxygen. A simple test like the gas pack can be performed to figure out if growth is possible without oxygen is doable. My unknown bacteria needs oxygen to grow. Throughout my study, the unknown bacteria was tested to see if it can grow in acidic conditions, however no growth occurred. Before inoculation, the substrate was a clear yellow color and in liquid state. After receiving the negative result, I kept the broth in the 37C incubator for 7 more days to confirm the negative result. Using premade slants with different types of sugars such as Mannitol, Sorbitol, Lactose, Trehalose, Maltose and Sucrose, to determine if the bacteria can metabolize glucose and if the bacteria is oxidative or fermentative. It was determined that my bacteria is strictly oxidative (needs oxygen to grow) and cannot metabolize glucose. Another test was done to confirm if the bacteria was able to utilize different carbohydrates in the presence of oxygen. I used Cellobiose, Arabinose, Adonitol, Fructose and Malonate wee tabs. Out of the 5 different types of carbohydrates, only 3 different
A mixed culture of two unknown bacteria was provided by the instructor. The methods used for
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
In a laboratory setting, it often becomes necessary to identify an unknown organism. In this experiment, researchers classified an unidentified bacterium based on its physical structure, colony morphology, optimal conditions and metabolic properties. A Gram stain using crystal violet, iodine, and safranin and a simple stain using methylene blue characterized the organism’s cell wall. Cultural behavior was classified by inoculating the organism onto nutrient agar and incubating it at 37° C for 48 hours, and observing its behavior, as well as using SIM medium to test for motility. Optimal growth temperature was
The purpose of this lab was to identify two unknown bacteria cultures using various differential tests. The identification of these unknown cultures was accomplished by separating and differentiating possible
The purpose of this lab was to identify two unknown bacteria cultures using various differential tests. The identification of these unknown cultures was accomplished by separating and differentiating possible bacteria based on specific biochemical characteristics. Whether the tests performed identified specific enzymatic reactions or metabolic pathways, each was used in a way to help recognize those specifics and identify the unknown cultures. The differential tests used to identify the unknown cultures were oxidase, catalase, lactose and sucrose fermentation, Kugler/iron agar, nitrate reduction, gelatin hydrolysis, starch hydrolysis, manitol salt, MR-VP, citrate, bile esculin,
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
(Harley, 2011, pp. 102-103) Sucrose and lactose serve as a fermentable carbohydrate sources which promote its growth of fecal forms and provide color change differentiation. (Harley, 2011, p. 104) Therefore, if E. coli is being platted on the EMB, after plate’s incubation period, it should produce a green metallic sheen on the agar due to the bacteria being a strong fermenter. My unknown bacteria tested positive for growth, and agar was fermented reddish burgundy in color. Subsequently, the unknown bacteria was later inoculated with a sterilized loop into the liquid tryptic soy broth and incubated at an appropriate temperature. Its results were used for proper identification of turbidity, spots and flocculation. (BD™ Tryptic Soy Broth (TSB), 2008) The results of the unknown were cloudiness and some settlement on the bottom of the tryptic soy liquid. The next step was conducted to find out if all the bacteria, as well as the unknown culture, required oxygen for growth, varying from an aerobic environment, where bacteria needs oxygen to grow, to facultative anaerobic environment, where bacteria will grow either with or without oxygen but better in its presence. All the bacteria, along with the unknown, were separately inoculated and tested. My unknown culture results tested positive for growth in facultative anaerobic environment. In the next sequence of lab experiments, the results of unknown bacteria were determined by glucose fermentation,
Escherichia coli is a Gram-negative rod that was tested on ten different biochemical testing procedures. The Escherichia coli culture on the MacConkey agar had growth and experienced fermenting of lactose (color change to yellow). This is an expected result because this agar only grows Gram-negative bacteria so it is a selective media. The bile salts and crystal violet are the agents that make the agar select against Gram- positive species. The pink tint is probably from the precipitation of bile salts. The Bacillus subtillis is a Gram-positive bacterium so it was not supposed to show growth or fermentation, but it showed growth without a color change (no lactose fermentation). This was due to the over streaking of maybe Escherichia coli or Enterococcus durans, which are Gram-negative species. Staphylococcus epidermis had a growth along with a yellow to orange color change. Media too was clear.
Abstract The purpose, or objective, of this lab was to identify an unknown bacteria by conducting different tests. Some of these include: oxidase test, fermentation tests, and catalase test. DNA fingerprinting was also conducted on the unknown sample, in order to provide another accurate means of identifying our DNA sample. The main objective for the identification lab was to be able to use known methods to identify our bacteria, including being able to follow a flowchart to aid in the process.
Life History and Characteristics: Staphylococcus aureus is a gram positive bacterium that is usually found in the nasal passages and on the skin of 15 to 40% of healthy humans, but can also survive in a wide variety of locations in the body. This bacterium is spread from person to person or to fomite by direct contact. Colonies of S. aureus appear in pairs, chains, or clusters. S. aureus is not an organism that is contained to one region of the world and is a universal health concern, specifically in the food handling industries.
2. Introduction: Each student was given unknown bacteria and was instructed to perform a variety of experimental tests that would help to identify their bacteria. During the process of identification, the unknown bacteria was added to many different testing medias using aseptic technique. They are as follows: lactose fermentation on eosin methylene blue (EMB), TSI (Triple Sugar Iron agar), Phenol red sucrose, the SIM test, H2S by SIM, IMViC (indole, motility, voges-proskauer, and citrate), Urease (urea broth), PDase (Phenylalanine Deaminase), Lysine Decarboxylase, and Ornithine Decarboxylase. Colonial morphology on EMB was used to
Introduction: Through the conduction of numerous experiments, the identity of two bacterial isolates was determined. The tested specimen was an unknown sample of a mixed culture of two different species of bacteria. The first step that was taken was obtaining a pure culture of each species of bacteria by isolating one species from the other. Once isolation was complete, the isolated cultures were tested using procedures that had been performed during previous lab sessions. A gram stain was performed on the two isolates. The isolate which had tested gram negative was then tested for the presence of cytochrome C and lactose fermentation. For the gram positive isolate, cell shape was determined and a catalase test was performed.