Identifying Fecal Metabolites in fly guts via Liquid Chromatography Mass Spectrometry

Blow flies assist in the transmission of disease-causing microorganisms also known as pathogens. Blow flies help transmit pathogens by first interacting with an organism’s fecal waste and then interacting with living organisms to some extent. This interaction can take place through many ways such as direct contact or through things like foods and surfaces. An invasive blow fly species in Tennessee has been determined to be a health hazard to humans. This project focuses on measuring fecal metabolites in blow fly guts by DNA extraction and liquid chromatography mass spectrometry. This research is important because it will allow researchers to better investigate and understand the transmission of pathogens from fly to human. Previous work by this group used phenol chloroform isoamyl solvent-based DNA extraction which is time-consuming and yields DNA unfit for advanced sequencing. This project proposes use of a kit-based DNA extraction method called Qiagen to yield clean DNA that can be further sequenced. All parts of extraction material are being examined for fecal metabolites including urobilins using an already validated liquid chromatography mass spectrometry method. It is expected that the fecal matter will be detected in a step of the DNA extraction process. When detected, wild type fly samples will be analyzed for the presence of fecal matter and compared to its sequenced DNA. This research will help confirm the viability of Qiagen DNA extraction method to yield sequence able DNA and still be able to detect harmful fecal matter metabolites.