Puparial remains as historical and forensic indicators

Calliphorid puparia are highly resilient, and can persist for thousands of years. Their value can help determine mortuary behaviour, seasonality of death, relocation of the body, and possibly result in repatriation of remains. This relies on accurate identification of puparia but can prove difficult if the samples are damaged or lacking important taxonomic features. The durability of mitochondrial DNA allows the application of DNA-based identification methods to insect samples discovered from archaeological excavations, in particular puparia and remaining arthropods. The aim of this study was to use multiple molecular techniques to examine the ability to extract and sequence small fragments of DNA from aged puparia for the purpose of identification. Puparia from 300 year old mummified human remains, and a Peruvian mummy bundle of 500-1200 years old were obtained. A modified phenol:chloroform:isoamyl extraction technique was used on both single and pooled puparia and nested PCR performed to amplify a 228bp region of the cytochrome oxidase I (COI) gene. 454 sequencing was also used on both sample sets. Phylogenetic analysis was applied to determine identity. In both cases, potential species identities were made. This study has demonstrated the potential for the use of multiple molecular techniques in identification of puparial remains of various ages to aid in both historical and forensic cases.