Student Reflection - John Pinard

John Pinard during his summer researchThis summer I was the molecular biologist of the group. My duties were to take samples, data, and results from the team and analyze them on a genetic level. With each trial, I would take samples of the fish microbiome or test media and use a method of genetic analysis commonly used in biomedical research, RISA (Ribosomal Intergenic Spacer Region), to analyze our data. The advantage to using this method opposed to conventional methods (i.e. plating/culture) is that the method is limitless. RISA shows all bacteria communities, with conventional methods many communities are lost due to bacteria not growing on culture plates, or are outcompeted by neighboring communities.

Along a strand of bacterial DNA there is a section of nucleotides named the ISR (Intergenic Spacer Region). This region is highly variable among bacterial species, which makes it an appropriate model to interpret bacterial community compositions. With our samples retrieved from the experiments I extracted DNA, accomplished PCR (thousands fold amplification of the ISR), then ran agarose gel electrophoresis. In the agarose gel electrophoresis, electricity streams these ISR sections of DNA across the agarose gel leaving these bands implanted in various locations along the gel according to size. The gel is then imaged with computer-assisted UV light imaging software. Since each band represents a different bacterial community, I see when and how much a community is altered by our experiments through the differing banding patterns.

In addition to the experiments detailed in our FAST proposal, I also completed other experiments to provide further data that will be used later. One was anaerobic vs. aerobic bacteria prevalence. Another was trying different test media to see which showed closest representation to the actual fish skin microbiome. All are very important data in order to simulate the normally fish skin microbiome.

This past summer, in addition to working on the FAST projects, I worked as a veterinary technician in a rural town in the Texas panhandle. I assisted veterinarians in large and small animal surgery, diagnostics, hospitalizations, and herd health. My favorite and the closest to my FAST projects was the herd health aspect of veterinary medicine. Dr. Zach Smith DVM sees about twenty feed yards on a weekly basis. His duties include prevention and treatment of diseases within the Texas and New Mexico cattle feed yards. While employed Dr. Zach and I completed three necropsies on cattle who had potential Salmonella infections. We used similar laboratory techniques that I used in the FAST projects out in the feed yard. We removed sections of organs, and completed antibiotic sensitivity tests on the microbiome in order to treat the cattle. It was great to participate in real world applications of the methods and techniques learned through my FAST project.

A Doctor of Veterinary Medicine is my all-time goal and as mentioned previously the FAST projects gave an immersion in the field which I most want to pursue. I thank the FAST committee for the great experiences and knowledge I accrued this summer.