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Transcript
Exploring Microorganisms Around You By Marianelly Lopez, Edima Akpan, and Kelechi Okereke Microbiology is simply the study of microorganisms that are in every place imaginable. The importance of understanding microbial diversity and obtaining information is to find the origins of how viruses and diseases develop. Microbial diversity must be understood in order to use the knowledge of microorganisms to find cures for diseases and viruses. In order to be a microbiologist, one must be curious about small microscopic life, study microorganisms from various environments, and be rigorous in the study of microbiology. The domains of the old tree of life used to be Prokaryotes and the Eukaryotes. But in today’s society of science, the new domains are Archaea, Bacteria, and Eukarya. Archaea and Bacteria are no longer grouped in the same category under Prokaryotes because their structures are different from one another. Bacteria have peptidoglycen in its cell membrane made up of phospholipids while Archaea has a polysecondary membrane. In addition, Eukarya contains a nucleus whereas Archaea and Bacteria do not. Today’s three domains of life are based upon the 16S ribosomal unit to classify microorganisms because it is the most conserved subunit. The small subunit is used because of the nucleotide differences in the ribosomal unit to identify groups of species. Some of the techniques that we used in this scientific experiment are Fluorescence in Situ Hybridization and inoculation. Procedures that we used were culturing and isolating. The objective of this experience was to study the microorganisms in the environment and examine how diverse they were. The purpose of this experiment is to be able to grow microorganisms in an enclosed area and to be able to identify the microorganisms’ classification. The first part of our experiment was to collect microorganisms from various places. Samples were taken from the Memorial Pond, the underside of a shoe, and the last from the refrigerator. The microorganisms had to be placed on agarose gels containing media with nutrients, and were incubated for about two days. After the incubation, we saw the results from under a dissection microscope. Edima’s sample contained homogeneous colonies, each being pinkish and smooth. Marianelly’s sample seemed to contain various different colonies. Some were veinshaped and white; others were more pinkish or yellowish. Kelechi’s sample had two distinct colonies. One colony was small, thin, clear, and tightly packed together, while the other colony was white and separated from each other. That was only the first part of the whole experiment. The next step was to collect a specific colony from the previous experiment. Using the techniques inoculation, incubation, and striking, we grew our chosen colonies on agarose gel containing media (food) with nutrients for one day. All of our results were different. Kelechi’s Petri dish was covered with the chosen organism. They were exactly the same ones that he had collected before. His colony contained white splotches, had ridges and smooth sides, and were more whitish than the agarose gel. Edima’s Petri dish had one colony following the streak that she had made earlier. This colony was pinker than the original colony, and contained smooth pinched colonies. All of her colonies were tiny circles. Marianelly’s Petri dish seemed to have grown two different colonies in the agarose gel. One of the colonies were a pale pink color and were separated from each other as well as the other existing colony, while the other colony was a dull white and were tightly formed together and following the streaked line. The final part of the experiment was to identify which colonies were bacteria, Archaea, or eukaryotes through a technique classified as Fluorescent In Situ Hybridization (FISH). This powerful technique evaluates and analyzes the presence of organisms in a community, their phylogeny, morphology, and number by targeting the 16S rRNA. To differentiate the organisms, fluorescent probes were added to the samples. If the sample has red organisms, it is universal bacteria. If it has blue organisms, it will either be Archaea or Actinobacteria. If the sample turns green, it will either be universal eukaryote or Gama proteobacteria. Results from this portion of the experiment were interesting. All samples contained universal bacteria as observed using the fluorescent microscope, given by the red color. Kelechi’s sample contained specifically Gama proteobacteria given by the green color, while Edima and Marianelly’s samples contained Archaea and Actinobacteria, with the assigned color blue. To sum things up, we saw how diverse our samples were. Some of the samples were similar, while others were different. We accomplished our goals for the project. We explored microbial diversity, were able to grow microorganisms in a lab, and were able to identify the microorganisms’ location on the tree of life. Overall, the experiments were successful. Each of us SMASH scholars enjoyed our experience. We all enjoyed doing the project and it has made us more interested in pursuing careers in science. We have learned how many diverse microorganisms are around us and how there is a newer version of the tree of life. As Marianelly said, a member of the team, “…everything that we did was exactly what I had imagined what we would do and more.” This project has really helped us to experience what it was like to be working in a real lab. We were able to learn a lot in terms of microbiology. The project has overall been an enjoyable experience and we all had a really great time conducting this project. We can only hope that we could do it again. REFERENCES: 1. Madigan, M.T., Martinko, J., Parker, J., Brock Biology of Microorganisms. 12th ed. 2003, Upper Saddle River, NJ: Prentice Hall. 2. "Main Page." Wikisource, The Free Library. 20 Jun 2009, 06:18 UTC. 4 Jul 2009, 05:10 <http://wikisource.org/w/index.php?title=Main_Page&oldid=257433>. 3. <http://microbewiki.kenyon.edu/index.php/MicrobeWiki>. 4. Philip Hugenholtz, Gene W. Tyson, and Linda L. Blackall. Design and Evaluation of 16S rRNA-Targeted Oligonucleotide Probes for Fluorescence in Situ Hybridization. Methods in Molecular Biology, vol. 176: Steroid Receptor Methods: Protocols and Assays. Chapter: 6-Hugenholtz.