Survey of Bacterial Diversity Associated with Various Life Stages of Lucilia sericata and Phormia regina Collected from Central Virginia

Reliable and valid methods for the postmortem interval determination are frequently sought after. Information obtained from the colonization of insects and the related bacterial succession on carrions may be utilized for PMI estimation but this can lead to erroneous results due to known variations between individuals and species. Apart from the bacterial communities found with human remains, it is important to note what bacteria may be specifically associated with blow flies colonizing these immediately after death. Blow flies often associated with carrions in Virginia are Lucilia sericata and Phormia regina. This comparative analysis of the various life stages of Lucilia sericata and Phormia regina was performed as the initial setup of an ongoing study including various species of blow flies associated with human remains in central Virginia. Although extensive data exists on the biology of blow flies, a well-replicated study on bacteria associated with different life stages of many blow flies is missing. In this study, we characterized bacteria associated with different life stages of Lucilia sericata and Phormia regina by using 16S rDNA MiSeq™ sequencing. Five Lucilia sericata colonies and four Phormia regina colonies were established from female flies collected from Richmond, VA and nearby areas on beef liver bait. First generation eggs, third Instar larvae, pupae, adults, and second generation eggs were collected from each colony for DNA extraction using organic CTAB extraction method and dual-index 16SrDNA MiSeq™ sequencing using the protocol as described by Kozich et al (2013).