New approach to application of mid-infrared photoacoustic spectroscopy in forensic analysis: Study with the necrophagous blow fly Chrysomya megacephala (Diptera: Calliphoridae).

Insects can provide clues in a variety of ways to assist in criminal investigations. The FTIR-PAS technique has been successfully used to assess the cuticular chemical profiles of insect samples from different groups and for several goals. However, until now, it has never been used to evaluate samples of forensic interest, despite providing faster results, compared to the methods currently used. In this study, mid-infrared photoacoustic spectroscopy was employed to assess the cuticular chemical profiles of different stages of development of the blow fly Chrysomya megacephala sampled from two distinct populations. The results showed that this technique enabled detection of significant differences between the main vibrational modes of the chemical bonds present in the cuticles of the two populations and the different stages of development of the fly. The method enables identification of the age of individuals collected at the crime scene, as well as the distinction of different populations. Therefore, this methodology could assist in forensic investigations, in both estimating the Postmortem Interval and determining the location where the crime occurred, or whether the body had suffered some type of translocation. The technique provides high reproducibility and fast analysis, so its application for analysis of C. megacephala is a viable option in forensic crime investigations.

Hydrocarbon and Fatty Acid Composition from Blowfly Eggs Represents a Potential Complementary Taxonomic Tool of Forensic Importance.

One of the most important contributions of forensic entomology is to assist criminal expertise to determine the postmortem interval, which depends on the duration of the immature stages of insects of forensic interest. On the other hand, the time of development of the different stages varies according to the species; therefore, its identification is essential. Currently, few studies have investigated the use of cuticular hydrocarbons, and none regarding fatty acids, as complementary taxonomic tools to expedite species identification. Therefore, we evaluated whether cuticular hydrocarbons together with fatty acids of eggs of flies of the family Calliphoridae, main group of forensic interest, can be used to distinguish species. The analyses were performed by chromatographic techniques. The results show that there are significant differences between the composition of cuticular hydrocarbons and fatty acids between species and, therefore, they can be used to provide a complementary taxonomic tool to expedite the forensic expertise.

Effect of temperature on the chemical profiles of nest materials of social wasps.

Social insects depend on their nests for protection against predation and abiotic threats. Accordingly, the chemical compounds present in the material wasps use to build their nests can both facilitate communication and repel predators. It is herein hypothesized that different wasp species build their nests with different structure and substrate materials and that such materials consist of chemical compounds related to unique wasp behavior and outside temperature variation. To test this hypothesis, nests were collected from three species of social wasps, the samples of which were subjected to temperature variation under laboratory conditions. The compounds present in the substrate were analyzed by gas chromatography coupled to mass spectrometry. Chemical compounds identified in the nest material of the three species responded differently to temperature variation. Chemical compounds from Polybia nests were altered significantly when subjected to temperature variation, whereas the nests of Polistes versicolor did not significantly change in relation to the control. The differences found between Polistes and Polybia nests may be related to genetic factors, but also to the type of nest they construct. It is possible that divergent evolutionary strategies for maintaining colony temperature, as a function of the chemical composition of the nests, may have appeared between wasps that have open and closed nests. In relatively small colonies, nest substrate is more resistant to temperature variation because it is composed of a greater diversity of elements and thus capable of holding heavier, longer carbon chains. Our results suggest that chemical compounds in the nest material of the three wasp species analysed responded differently to fluctuating ambient temperatures and that such variation could result from the biochemical differences of unique wasp species or from thermoregulation strategies of colonies.

Effect of temperature on survival and cuticular composition of three different ant species.

Climatic factors, such as temperature variation, interfere with the survival of insects. To respond to these variations, insects have some specific characteristics. These include water content of the body, thickness of the lipid layer, as well as the qualitative and quantitative characteristics of cuticular chemical components. This study hypothesizes that different ant species respond to temperature changes in different ways and that such differences may be associated with cuticle hydrocarbons (CHCs) and fatty acids. As model ant species, Atta sexdens, Odontomachus bauri and Ectatomma brunneum were used for experimental analyses. Ants were submitted to a water bath for 5 h at different temperatures, and their CHCs and fatty acids were identified and quantified, followed by correlating these chemical compounds with temperature variations and the survival. Temperatures below 30 °C did not affect the survival of the three species. E. brunneum had a higher percentage of survival at temperatures above 30 °C. O. bauri was the most sensitive species with 100% mortality at 40 °C. Survival was found to be unrelated to any of the identified fatty acids. However, CHCs underwent significant quantitative and qualitative variation, as shown by an increased percentage of CHCs with longer chain length of linear alkanes at temperatures above 30 °C. These increase enables these ants to maintain the integrity of their cuticle and survive at temperatures above 30 °C. It can be concluded that the forager ants studied respond differently to temperature variation and that changes in the conformation of CHCs are in line with the ecological characteristics of the different studied species because, they vary in terms of diurnal/nocturnal foraging and types of environments foraged. Among the three species, E. brunneum foragers were found to be more active under adverse conditions and more tolerant to temperature variation with the correspondingly appropriate changes in CHCs composition.

New method for estimating the post-mortem interval using the chemical composition of different generations of empty puparia: Indoor cases.

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.

Chemotaxonomic Profile and Intraspecific Variation in the Blow Fly of Forensic Interest Chrysomya megacephala (Diptera: Calliphoridae).

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.