A series of biochemical tests were conducted in order to determine the identity of an unknown gram negative bacterium. The unknown had the potential to be one of six different gram negative bacteria: Escherichia coli, Pseudomonas aeruginosa, Enterobacter aerogenes, Proteus mirabilis, Klebsiella pneumonia, or Salmonella typhirmurium. After using the T-streak method to isolate pure colonies and confirming the gram negative nature of the unknown bacterium the unknown was subjected to seven biochemical tests and the results compared to the results of the six known bacteria for the same tests. After performing the Triple Sugar Iron Agar Test, Sulfur Indole Motility Test, Methyl Red Test, Voges-Proskauer Test, Citrate Test, Urease Test, and Gelatin Test it was confirmed that identity of Unknown #15 was Salmonella typhirmurium.
While all bacteria retain basic qualities identifying them as prokaryotes they do not function, grow, or even metabolize in the same way. These differences distinguish bacterial species from one another, like a thumb print identifies a single individual from every other person on earth. Testing for these unique factors through a series of biochemical tests and staining techniques allows for an unknown bacterial strain’s “thumb print” to be generated. From there the unknown’s thumb print can be compared to the thumb print of known bacterial species allowing for identification.
The first step in identification is performing a Gram Stain. Due to the fact
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 first step toward identifying this unknown organism was to perform a Gram Stain to differentiate between gram positive and gram negative bacteria. This is an important step because it directs what the next tests will be. My Gram Stain on sample #12 showed that the bacteria was gram negative, however, after receiving the results of the OF glucose, H2S, Citrate, Urease and Motility tests, it was apparent that my Gram Stain was contaminated. I then performed a catalase test which came back negative, so I ordered a Bacitracin disc, Optochin disc and a CAMP test which had to be incubated overnight. After receipt of those test results,
Bacteria are ubiquitous; they can be found on the skin, in the soil, and inside the body. Because of the very nature of this ubiquity, it is important to be able to determine between different strains of bacteria. An example of this is determining the causative agent for a disease so that the patient will be treated with the appropriate antibiotics. It may be important to determine the bacteria in a certain region, because like with enteric bacteria, it is normal to find them in the digestive tract as they are in a symbiotic relationship with our bodies in this area; however, they also cause opportunistic infections in places outside of the digestive tract to our detriment, such as with a urinary tract infection. Some strains of bacteria are common to nosocomial infections, and identifying these bacteria as such helps create the guidelines for healthcare workers in antiseptic technique. All of the morphology and characteristics of each strain of bacteria help us to better understand the role of bacteria in the body as well as helps us understand how they can cause illness, and what treatment regimen to set in place. In lab this semester, a sample of unknown
This lab experiment was done for the purpose of learning how to determine a gram negative bacterium based on multiple tests learned throughout the semester. My gram negative unknown bacterium given to me was Salmonella typhimurium based off of the following tests; Triple Sugar Iron Agar (TSIA), Sulfate Indole Motility (SIM), Methyl Red (MR), Voges-Proskaur (VP), Citrate, Urea Hydrolysis, and Gelatin Hydrolysis. Each test performed gives results such as motility, acid production, fermentation, carbon requirements, or detection of certain coenzymes. With a process of elimination, I determined which bacteria it was not and which bacterium I had, S. typhimurium. The expectation was to master the techniques for each test and utilize the results to determine the unknown bacterium I was given within a two-week period.
There are many reasons for knowing the identity of microorganisms. The reasons range from knowing the causative agent of a disease in a patient, so as to know how it can be treated, to knowing the correct microorganism to be used for making certain foods or antibiotics. This study was done by applying all of the methods that have been learned so far in the microbiology laboratory class for the identification of unknown bacteria. The identification process can be completed with a series of deferential stains and biochemical tests. Creating a dichotomous key helps to limit the amount of biochemical tests done on an unknown organism and by observation
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 main idea of this experiment was to correctly identify the unknown bacteria, #3. Identification of unknown bacteria yields multiple benefits in many different areas in the research of microorganisms. In this experiment I performed many different test dealing with things such as the presence of enzymes, fermentation abilities and different chemical reactions. Observations made from the tests were then compared to a gram negative unknown chart in order to identify the bacteria. Based off of my results and the chart, I concluded the bacteria #3 was the bacteria Escherichia coli. E. coli is most commonly found in the intestines of warm blooded organisms. Most E. coli strands are non pathogenic however, there are strands
As the flowchart shows, a series of tests were conducted to identify the unknown bacterium #65. Microscopic observation of the gram stain indicated a gram-positive coccus bacterium. S. epidermidis was used as the gram-positive control while E. coli was used as the gram-negative control. This observation led to the elimination of all gram negative and rod-shaped genera: Enterobacter, Citrobacter, Klebsiella, Escherichia, Pseudomonas, Serratia, Alcaligenes, Neisseria, Proteus, Salmonella, Shigella, Erwinia, Veillonella, Flavobacterium, Bacillus, Arthrobacter, Lactobacillus, Listeria and Kurthia (2). By performing the catalase test, it was determined that the bacterium was catalase negative and it did not produce bubbles. M. luteus and E. faecalis were used as positive and negative controls, respectively.
There are many reasons for knowing the identity of microorganisms. The reasons range from knowing the causative agent of a disease in a patient, so as to know how it can be treated, to knowing the correct microorganism to be used for making certain foods or antibiotics. This study was done by applying all of the methods that I have been learned so far in the microbiology laboratory class for the identification of an unknown bacterium.
A highly conserved gene will be used to identify a prokaryotic species isolated from the body. Fundamental lab techniques will be also explored and utilized, such as amplifying using PCR, cloning, and transforming the gene into a host cell. DNA electrophoresis and specific substrate plating will serve as analysis check points. The final product will be sequenced and compared to similar species to observe phylogenetic relationships.
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.
The purpose of this study was to identify the unknown bacterium using biochemical tests and various methods that had been learned from previous the microbiology laboratory class. Identifying the unknown bacterium was determined by separating and differentiating possible
Often scientists work with bacteria that do not come in a labeled test tube— for example, bacterial samples taken from infected human tissue or from the soil—and the scientist must then identify the unknown microorganism in order to understand what behavior to expect from the organism, for example, a certain type of infection or antibiotic resistance. However, because of the relatively few forms of bacteria compared to animals and because of the lack of bacterial fossil records due to their asexually reproductive nature, the taxonomy used to classify animals cannot be applied to bacteria (Brown 275). In order to classify unknown bacteria, a variety of physiological and metabolic tests are available to narrow a sample down from the fathomless number of possibilities into a more manageable range. Once these tests have been performed, the researcher can consult Bergey’s Manual of Determinative Bacteriology, a systematically arranged and continually updated collection of all known bacteria based on their structure, metabolism, and other attributes.
In the world of microbiology it is vitally important to be able to discern the identities of microorganisms. Not only is it important in a lab setting but as well as in healthcare in general. Properly identify what strain of bacteria a person has will aid in the proper medicine and dose given. Throughout the semester we have learned about different types of bacteria and certain test that can clearly identify them. The purpose of this lab report is to identify a Gram-positive or Gram-negative bacterium. Using all the knowledge of procedures and lab techniques identify the unknown and discuss all the tests you performed.
The Unknown Bacteria 36/Bacteria # 2 on a TSA plate was examined by the naked eye and under a dissecting microscope. Bacteria # 2 was approximately 3 - 4 mm in diameter. They were circular in form with an entire margin and a flat elevation. The colonies were rough (granular), translucent, and white brownish color with black granules. The Gram stain resulted in a Gram negative rod. After the Gram stain was completed, the bacteria were streaked on an Eosin -Methylene Blue Agar plate and an Enterotube II was inoculated.