Unknown Lab Report

The following tests according to the lab manual were performed: gram stain, fermentation tubes, methyl red, vogues proskauer, sulfur, indole, motility and growing it up on MacConkey agar. The gram stain was performed incorrectly the first time. This is because the decolorizer was not on the bacterium slide for long enough, giving a false outcome.

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.

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.

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.

Purpose: The purpose of this lab is to help you become a little familiar with some of the tests that can be typically performed in a clinical or research lab facility. These tests may help in determining a particular pathogen’s growth needs.

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 IMViC test is a series of different tests that differentiate between enterics. One is the Indole test. This test tells whether the bacterium possesses tryptophanase which is the enzyme that breaks down tryptophan into indole. The agar contains tryptic soy broth so if the bacterium contains tryptophanase, indole is produced. This production of indole is seen by adding Kovac’s reagent which causes a red ring to be seen at the top of the tube. The citrate test is also used to see which kind of products the bacteria make. It uses a green agar slant that contains sodium and ammonium phosphate. Bromythymol blue dye is late added as an indicator. Inoculation of the slant with a needle using a zig-zag then stab technique was used with the gram positive bacterium. Conversion of the medium to blue is a positive citrate result. All plates, slants, and broths were incubated at 37°C for 24‐48 hours.

Negative result is no color change with mineral oil on top of solution. This bacterium has +/- reaction on MAC plate. It could survive in an acidic environment, but it not able to ferment lactose. The bacteria grow on the plate, but did not change the color of the plate to bright pink. This bacterium has a +/- result on Bile Esculin Agar.

In the Voges-Proskauer test, I inoculated the tube with bacteria from my TSA plate and incubated the tube at 37 degrees Celsius for 3 days. After three days I placed some Barrits Reagent A and Barrits Reagent B in the test tube. The color change of red or pink indicates a positive reaction for acetoin which tells you that the organism is a butanediol fermentor.

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.

The main objective was to identify an unknown organism by utilizing skills we learned in our labs this semester. The purpose was to attain the possible identity of the unknown organism by actually performing biochemical tests and staining techniques we learned in lab. After performing and analyzing the results, we were able to use Bergey’s Manual of Systemic Bacteriology as a guideline to narrow down the genus of our organism test by test.

The objective of this experiment is to identify the organisms of two unknown bacterial cultures. Students must identify the species of the unknown bacteria by utilizing the techniques and information learned in previous laboratory exercises. These techniques include streaking for isolation, Gram staining, and specific biochemical tests. Students are given a map known as a dichotomous key, a guide in determining the identity of their unknown sample.

A wide variety of processes had been performed to determine what culture had been acquired in class. My group had acquired culture tube unknown #3. We first isolated the bacteria. In this step we took the broth of the unknown #3 and grew it on an agar plate. The first process that had been performed was discontinuous streaking, and continuous streaking to grow the culture for future use in the lab and to have extra to perform a whole variety of test to determine the unknown. The next experiment that had been performed

The identification of an unknown sample, particularly in the environment, can be the matter of life or death. Identifying an unknown environmental sample has countless of opportunities and benefits. Identifying pathogens in the environment can save the lives of thousands or even millions of people. Some bacteria are opportunistic pathogens such as Pseudomonas. These pathogens do not commonly infect healthy people, but rather take any opportunity possible such as an open wound or an immunocompromised system. Knowing the bacteria which can prevent or promote the growth of agriculture can help to feed populations. Being able to identify bacteria is a task not to be taken lightly. Many different tests are possible to determine a specific bacterium. Often it takes a combination of several different tests to narrow down and identify a specific genus and species.

In the case of this identification process, all of the bacteria were known to be gram negative. The first step in identification was to streak the mixed broth culture