Abai, MR

6th International Congress of DipterologyAvailability of the puparium as a DNA sourceSaigusa, K. (1), M. Matsumasa (1), Y. Yashima (1), M. Takamiya (2), & Y. Aoki (2)(1) Department of Biology, Iwate Medical Univiersity School of Liberal Arts andSciences, 3-16-1 Honcho-dori, Morioka, Iwate 020-0015, Japan(2) Department of Legal Medicine, Iwate Medical Univiersity School of Medicine, 19-1Uchimaru, Morioka, Iwate 020-8505, JapanIn forensic practice, fly puparia are often found on a corpse or coverturesuch as clothing even when the body is in advanced decomposition. Theseinsect remains can provide important forensic information, if their speciescan be properly identified, based on their ecological characteristics: e.g.,seasonal dominance, period of activity/diapause, habitat, etc. However, it isoften difficult even for entomologists to accurately identify insect speciesfrom the morphology of such shells. On the other hand, a molecularbiological technique that is simple and time-saving has recently beenapplied to identify the species of insects. We attempted to extract DNAfrom the puparia of two blowfly species (Lucilia sericata and Calliphoravicina) and evaluated the availability of the puparium as a DNA source.The blowflies were reared in a laboratory and the species were identifiedby the male genitarial shape of emerging adult flies. The puparia werecollected from a rearing container and the DNA were extracted using aDNeasy Tissue Kit (QIAGEN, Hilden, Germany). The 348 base pairs ofthe polymerase chain reaction (PCR) products of mitochondrialcytochrome oxidase gene subunit I (COI) were sequenced. The DNA wassuccessfully extracted and amplified from the puparia collected under dryrearing conditions. The mtDNA sequences of the PCR products extractedfrom the puparia were identical to those of the adults. Therefore, it wassuggested that the puparium would be useful for a DNA source. However,caution should be taken in preserving the puparia specimens, and avoidingDNA contamination by other organisms,as with other forensic DNAsamples for practical use.Key Words: puparium, mitochondrial DNA, species identification,postmortem interval, forensic entomology208