Discrimination of Diptera order insects based on their saturated cuticular hydrocarbon content using a new microextraction procedure and chromatographic analysis.

The nature and proportions of hydrocarbons in the cuticle of insects are characteristic of the species and age. Chemical analysis of cuticular hydrocarbons allows species discrimination, which is of great interest in the forensic field, where insects play a crucial role in estimating the minimum post-mortem interval. The objective of this work was the differentiation of Diptera order insects through their saturated cuticular hydrocarbon compositions (SCHCs). For this, specimens fixed in 70 : 30 ethanol : water, as recommended by the European Association for Forensic Entomology, were submitted to solid-liquid extraction followed by dispersive liquid-liquid microextraction, providing preconcentration factors up to 76 for the SCHCs. The final organic extract was analysed by gas chromatography coupled with flame ionization detection (GC-FID), and GC coupled with mass spectrometry was applied to confirm the identity of the SCHCs. The analysed samples contained linear alkanes with the number of carbon atoms in the C9-C15 and C18-C36 ranges with concentrations between 0.1 and 125 ng g-1. Chrysomya albiceps (in its larval stage) showed the highest number of analytes detected, with 21 compounds, while Lucilia sericata and Calliphora vicina the lowest, with only 3 alkanes. Non-supervised principal component analysis and supervised orthogonal partial least squares discriminant analysis were performed and an optimal model to differentiate specimens according to their species was obtained. In addition, statistically significant differences were observed in the concentrations of certain SCHCs within the same species depending on the stage of development or the growth pattern of the insect.

Ultrastructure of preimaginal stages of Piophila megastigmata McAlpine, 1978 (Diptera, Piophilidae): a fly of forensic importance.

Piophila Fallén, 1810 is a genus of small flies composed of two species: Piophila casei (P. casei ) (Linnaeus, 1758), worldwide distributed, and Piophila megastigmata (P. megastigmata ) McAlpine, 1978, recently referred in the Palaearctic Region, from the Iberian Peninsula. Both species share ecological niche and are very interesting for forensic purposes, since they are present in carrion in advance stages of decay and have been found to be related to human corpses. The immature stages of P. megastigmata have ever been described, so this paper gives the ultrastructural morphologies of all preimaginal stages of P. megastigmata studied by light microscopy and scanning electron microscopy (SEM). Particular attention is given to pseudocephalon features­antenna, maxillary palps, facial mask, etc.­cephalopharyngeal skeleton, anterior and posterior spiracles, tegumentary sculpturing, and anal division among others. A comparative analysis of the main distinguishing features is made in order to understand how those features evolve along the developmental process, while larvae II and III are morphologically similar to each other, the larva I shows particular features. Larvae of all stages and pupae are easily distinguishable from other Diptera of forensic importance just based on the presence of trichoid sensilla associated to respiratory slits, instead of peristigmatig tufts, as well as on thewell-known disposition of anal papillae. The shapes of both dorsal edge at the basal part of mouthhook and dorsal bridge of cephalopharyngeal skeleton, and the tegumental ornamentationmay be considered as good features to distinguish the Piophila species, especially for P. megastigmata and P. casei . At the SEM level, shape, number, and arrangement of oral combs, oral ridges, sensilla of maxillary palpus, papillae of anterior spiracle, scales of spiracular field, and posterior spiracles represent good features to distinguish P. megastigmata from P. casei, but further studies will be necessary in West-Paleartic specimens of latter species. The key for identifying third instar larvae of forensically important Piophilidae in the Iberian Peninsula has been updated to include P. megastigmata.

Estimation of postmortem interval in real cases based on experimentally obtained entomological evidence.

Using the entomological evidence obtained in several forensic cases analyzed in our laboratory for comparison, we evaluated the results of an experimental study carried out in a semiurban setting to determine the structure of the sarcosaprophagous fauna from a Mediterranean region of SE Spain. In all, 18 orders of arthropods were collected. The summarized experimental results refer to the most important taxa for estimating the postmortem interval. Thus, the seasonal character of certain Diptera species, such as Phaenicia sericata, Calliphora vicina, Chrysomya albiceps and Musca domestica, is cited. Among the Coleoptera, the role of Dermestidae as a necrophagous species, and of Staphylinidae and Histeridae as necrophilous, or Cleridae, Tenebrionidae and Nitidulidae as omnivorous, as well as their appearance on the corpse, is described. Among the Hymenoptera, Formicidae were as the most abundant group, acting as omnivores and not apparently related to any particular decomposition stage. The real cases are discussed using data from the literature and the experimentally obtained results. In every case, the most relevant factors for estimating PMI are briefly discussed, mentioning, when possible, the relation with the experimental results. We confirm the significance of the experimental results because they seem to be applicable to actual forensic cases, the details of which enlarge our very little knowledge on the subject in the Iberian Peninsula. The importance of regional faunistic studies of the sarcosaprophagous arthropod community, the results of which may be applied to forensic practice, is mentioned.

Studies on seasonal arthropod succession on carrion in the southeastern Iberian Peninsula.

A global study of the sarcosaprophagous community that occurs in the southeastern Iberian Peninsula during all four seasons is made for the first time, and its diversity is described with reference to biological indices. A total of 18,179 adults and, additionally, a number of preimaginal states were collected. The results for the main arthropod groups, and their diversity are discussed in relation to the season and decompositional stages. The results provide an extensive inventory of carrion-associated arthropods. An association between decomposition stages and more representative arthropod groups is established. With respect to the biological indices applied, Margalef's index shows that the diversity of the community increases as the state of decomposition advances, while Sorenson's quantitative index shows that the greatest similarities are between spring and summer on the one hand, and fall and winter, on the other.

Protocolo para la recogida, conservación y remisión de muestras entomológicas en casos forenses / Collecting, keeping and shipment protocol for entomological evidences in forensic practice

Dada la importancia y creciente utilización de las evidencias entomológicas en la práctica forense, este trabajo propone un protocolo para la adecuada recogida, conservación y remisión de muestras entomológicas para su inclusión en la rutina médico-legal.La utilización de este protocolo permitirá la obtención de una base de datos homogénea y per-fectamente extrapolable a otros casos en el área geográfica (AU)