D3158: Metagenomic analysis of Aedes aegypti viromes

Developments in metagenomic sequencing have opened the door for categorizing microbial ecology across many species. In Houston, the high prevalence of mosquitoes necessitates a deeper understanding of viral communities that both live in the environment and potentially pose a public health risk. Traditional culturing techniques have been insufficient for capturing accurate profiles of viral biomes of each species. We used a metagenomic approach to categorize the viral populations within Aedes aegypti by sequencing entire genetic material present in each sample. After collecting mosquitoes from various locations throughout Houston, we separated them by both species and sex using large scale morphology as well as DNA barcoding. Total DNA and RNA was extracted from samples, RNA quantities were converted to cDNA and the cDNA/DNA combinations were sequenced using 454 technologies. We used a modified BLAST workflow developed by our team to analyze the results. Our first analysis looked at both males and females and indicated the presence of both RNA and DNA viral sequences in addition to thousands of unknown or potentially novel viruses. 51 different species of viruses were identified in females and 40 in males. The female samples carried more RNA viruses than the male, however further pools are needed for comparison before final conclusions can be drawn. Currently, we are in the process of building libraries from new samples including other species typically found in our area such as Culex quinquefasciatus and Anopheles gambiae. These studies will contribute to our understanding of viral diversity.