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Identification of Bacteria by Enzymatic Activitiy Courtney Ballinger Department of Biology, Tennessee Technological University, Cookeville, TN Introduction Bacteria are powered by biochemical reactions known as metabolism (Tondo et al 2004). The main purpose of metabolism is to generate energy and use it for the work of the bacteria (Kilian 1978). Metabolic reactions are performed by enzymes and with each individual step throughout the metabolic pathway require the use of a different enzyme (Krivobok et al 2003). Different bacterial species use different pathways (King et al 1978). Complement of enzymes found in a particular bacterium can be used as a fingerprint to identify it (Goss 2006). Objective and Hypothesis The objective is to be able to identify various bacteria so the identification of an unknown bacterium is possible. The hypothesis is that I will be able to identify an unknown bacterium using six enzyme tests on less than ten organisms. The null hypothesis would be that the unknown bacterium will be identified by having to use more than ten organisms for the identification of the unknown bacterium. Methods and Materials Identification will be achieved by oxidation-fermentation tests, catalase and oxidase tests, staining, and culture methods in the lab using methods outlines in the Biology 3230 Lab Manual (Goss 2006). Materials needed will be two starch agar plates, two DNA agar plates, two oxidation-fermentation glucose tubes, oxidase test strips, hydrogen peroxide, slides, sterile wooden sticks, sterile mineral oil, and Gram stain reagents. Results were recorded in Table 1 after each experiment was conducted. Results The identification of various unknown bacteria using this method was successful. The graph (Figure 1) shows the success rate of identifying unknown bacteria by this process. My results proved that my hypothesis of being able to identify unknown bacteria using only six enzyme tests on less than ten organisms can be accepted. The null hypothesis of having to use more than ten organisms can be rejected. The results supported the results found in the literature sources used. Conclusions Overall the results that I collected positively correlated with the previous work and information that researchers in the past have collected on this topic. Fast identification of unknown bacteria is possible with the assistance of enzyme reactivity. Using the identification of other bacteria and enzyme reactivity, it allows unneeded tests to be eliminated from the normal routines commonly used. This procedure of identification can benefit people throughout the medical field. With the assistance of enzyme tests this method could save time, money, and possibly lives. Acknowledgements I would like to thank Dr. Goss for allowing me to use her laboratory to perform my experiment. I would also like to thank those in the TTU faculty who helped me in my research and ideas. Finally, Suzanne Ballinger, John Crouch, and Nikki Talbott deserve a great amount of thanks for critiquing my manuscript. Figure 1. Comparison of Tests
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