The Difference in Microbial Diversity Between Moving Water and Still Water Cecilie Elliott and Tatianna Griffin Department of Biology and Environmental Sciences, Longwood University Collection Date February 8th, 2017 (goes on blog) Introduction Methods The objective of this experiment is to get a small glimpse into the microbial diversity of Prince Edward County by testing samples taken from and around Buffalo Creek, behind Longwood University’s Environmental Education Center (EEC) and from sites at the campus of Hampden-Sydney College. In order to compare a lot of the gene sequences of the variety of microbes, scientists in other studies normally use the 16S rRNA gene clone libraries as a way to provide a reliable and valid estimate. Figure 6. Gel Electrophoresis for undiluted and diluted samples. The figure shows the separation of the DNA sequence fragments by size. The left figure is the top gel. The right figure is the bottom. Table 2. Top alignments for BLAST matches. The DNA sequences were searched in a BLAST database to find the closest match for bacteria identification. Results A Figure 1. Comparison of library size to the number of observed phylotypes in bacterial libraries. Libraries are grouped into environmental categories, both aquatic or non-aquatic[1] Conclusions B The results do not correlate with the initial hypothesis stating that more microbial diversity is found in the soil of running water than in still water. Standing water and flowing water have similar microbes living and growing in the soil in Prince Edward County, VA according to the bacteria Identified. Bacillus thuringiensis helps to make plants and crops resistant to insect attacks [2] Pseudomonas koreensis was found to have some sort of biocontrol ability against other microbes and allows it to protect against infestation [3] Pseudomonas azotoformans was found to act as an herbicide degrader [4] The soil portion of our samples contained more microbes that the actual water itself Specific Aim The goal of this experiment is to investigate whether the soil of running water or the soil of still water will have more microbial diversity Still water should have more diverse microbial communities due to the fact that running water is constantly flowing over the sediment and moving it along with the tide of the water. Figure 4. Number of bacterial colonies over time. (a) All the samples were incubated at room temperature for 24 hours and 48 hours. (b) Photographs of the growing colonies on the undiluted plates after 48 hours. Table 1. Characterization of bacterial colonies. Certain characteristics of the bacteria were observed from each agar plate. Future Directions With these findings, crops can be genetically engineered with a specific strain of microorganisms that are safe for humans and safe for the environment. This means that insects cannot ruin crops and harmful pesticides can stop being used; meaning that they will no longer hurt the environment, affect humans, or create pollution in the air, water, and soil. This experiment could be used in future experiments to discover the exact reason these bacteria are located in Prince Edward County, when they originated from other distinct places. Figure 2. Two of the sample sites.The image to the left shows the pond sample sight behind the Longwood Environmental Education Center. The image to the right presents the site at Buffalo Creek where some of the samples were also collected. B) C) A) E) D) References Joshi AA, Kanekar PP, Kelkar AS, Shouche YS, Vani AA, Borgave SB, Sarnaik SS. 2007. Cultivable Bacterial Diversity of Alkaline Lonar Lake, India. Microbial Ecology; 55(2): 163-172. Mohamed IA, Griko N, Junker M, Bulla LA. 2010. Bacillus thuringiensis: A genomics and proteomics perspective. Bioeng Bugs; 1(1): 31-50. Haiyan L, Shikai H, Ruifang L, Ping C, Changwei G, Bo Z, Longbiao G. 2016. Genome Sequence of Pseudomonas koreensis CRS05-R5, an Antagonistic Bacterium Isolated from Rice Paddy Field. Frontiers in Microbiology; (7): 1756 Nie ZJ, Hang BJ, Cai S, Xie XT, He J, Li SP. 2011. Degradation of cyhalofop-butyl (CtB) by Pseudomonas azotoformans strain QDZ-1 and cloning of a novel gene encoding CyB-hydrolyzing esterase. J. Agric. Food Chem; 55(11): 6040-6. Figure 5. Photographs of the specific colonies being isolated and analyzed. The specific colonies that will be sequenced are circled in blue on each plate. A) HSC Pond 3 Soil B) HSC Stream 4 Soil C) LU Creek Soil D) LU Pond Soil E) HSC Stream 2 Soil Figure 3. Photographs of the sites where the samples were taken. The image to the left was the site at Buffalo Creek. The image to the right was the site at the Pond behind Lancer Park Apartments.