D3397: Investigating blow fly associated bacteria and viruses

Pathogen surveillance is a topic of foremost concern to medical, biosecurity and forensic fields, and the unique life history of blow flies in relation to these interests makes them an exciting tool for such surveillance efforts. . These flies will often utilize decomposing remains as a site for many biological processes including feeding and oviposition, making blow flies a rich source of information on the animal and microbial populations with which they interact. To study such novel surveillance methodologies, RNA libraries were prepared using three different methods, and data was compared. TraditionalmRNA and small RNA libraries from laboratory maintained fly colonies as well as ribosomal depletion libraries created from wild type flies were analyzed for the presence of bacteria, viruses, and other microbes. These results were then filtered for common microorganisms associated with decomposition, insect life history, and human health and agricultural relevance. Pathogenic bacteria including Bacillus anthracis, several phage, plant pathogens and insect pathogens were identified. We determined that ribosomal depletion methods result in identification of the largest number of unique taxa, followed by small RNA and then mRNA preparations. As read cutoffs were imposed, small RNA libraries were shown to have the highest number of unique taxa, with a minimum of 100 reads. The results of this study indicate that ribosomal depletion and small RNA library preparation methods applied to fly populations are a useful tool for surveillance of environmental microbes.