Document Type

Student Research Paper

Date

Spring 2019

Academic Department

Biology

Faculty Advisor(s)

Debra Wohl

Abstract

Fecal Matter Transplants (FMT) are an effective yet underutilized treatment for potentially life-threatening Clostridium difficile infections. Following antibiotic treatment, an imbalance between the types of colonic microbiota naturally present in a person’s gut may occur, allowing the opportunistic pathogenic bacterium C. difficile to proliferate and reach virulent levels. Despite a 90% success rate, and patient reports of immediate improvement, FMTs are approved only as a last resort due to strict US Food and Drug Administration restrictions, which label them as “experimental”. This study aimed to compare microbial preservation methods to determine the method with the least detrimental effect on the composition of stool microbes. Fecal matter samples, collected from dogs, were homogenized with either sterile deionized water or 0.85% NaCl. The homogenized mixtures were then partitioned for immediate DNA extraction or for preservation with or without 25% glycerol prior to -80°C storage. DNA extraction was also performed on samples partitioned and stored at -80°C after 3 weeks and after 10 months of storage. All extracted DNA was then subjected to PCR amplification and sequenced. A Phred score was used to assess quality of DNA sequencing. Based on alpha and beta diversity analysis, the water and glycerol treatment resulted in the least amount of change in taxonomic composition and proportions when compared to the sample prior to preservation. Information gained from this study could be used to further improve FMTs and help fuel FMT related research in hopes of attenuating FDA restrictions.

Notes

Senior thesis.

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