Necrophagous insects such as blow flies (Diptera: Calliphoridae) are considered crucial in forensic entomology. Identification at species level and determination of larval stage are the basis for estimation of postmortem interval (PMI). Insect evidence can also be used in the determination of crime scenes, since body displacement is common. The aim of this study was to determine the chemotaxonomic profile and intraspecific variability of the forensically important blow fly Chrysomya megacephala (F. 1794). Adults were collected in the municipalities of Dourados-MS (Brazil) and Rio Claro-SP (Brazil), and then transferred to the laboratory for oviposition and development of the immature stages. Chemical analysis of cuticular compounds was performed by gas chromatography. Cuticular chemical profiles varied significantly between the two populations, as well as between developmental stages, supporting the use of these compounds as a complementary tool to help identify the species and its stages, along with geographical variability. This could greatly accelerate forensic investigations, eliminating the need to allow the fly larvae to develop until adult stage in order to confirm the species identity and sample origin.

Most flies of forensic importance are in two superfamilies, the Muscoidea and the Oestroidea, with similar life stages including the puparium. Upon completion of metamorphosis the adult fly emerges from the puparium, leaving behind an exuvia that is of potential significance in forensic investigation. The empty puparium is a durable piece of entomological evidence lasting several years. Through the study of chemical compounds, specifically the hydrocarbons of these puparia, it is possible to identify the species, in addition to how long they have been exposed to weathering and for this reason, these parameters can assist forensic entomologists in estimating long-term postmortem interval (minPMI). In corpses that take a relatively longer time to decompose, insects may use the same corpses for several oviposition cycles. Therefore, the aim of this study was to develop a new method to determine the PMI based on chemical compounds of the puparia from different oviposition cycles of the fly Chrysomya megacephala. The chemical composition of 50 puparia from different cycles of oviposition were evaluated by Gas Chromatography–Mass Spectrometry (GC-MS). In total, 60 compounds were identified ranging from C18 to C34, 38 of those were common to all generations. Our results demonstrate that chemical profiles can be used to differentiate puparia collected from successive cycles, and therefore valuable in the estimation of minPMI.

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