Use of Lucilia Species for Forensic Investigations in Southern Europe S
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Available online at www.sciencedirect.com Forensic Science International 177 (2008) 37–41 www.elsevier.com/locate/forsciint Use of Lucilia species for forensic investigations in Southern Europe S. Vanin a,*, P. Tasinato b, G. Ducolin b, C. Terranova b, S. Zancaner b,c, M. Montisci b, S.D. Ferrara b, M. Turchetto a a Dipartimento di Biologia, Universita` di Padova, Via U. Bassi 58/b, 35131 Padova, Italy b Dipartimento di Medicina Ambientale e Sanita` Pubblica, Sede di Medicina Legale, Universita` di Padova, Via Falloppio 50, 35121 Padova, Italy c Unita` Operativa Complessa di Tossicologia Forense e Antidoping, Azienda Ospedaliera - Universita` di Padova, Italy Received 26 February 2007; received in revised form 2 October 2007; accepted 25 October 2007 Available online 20 February 2008 Abstract The aim of this study was to highlight the importance of evaluating entomological evidence in forensic investigations on a regional scale. To evaluate climatic, geographical and environmental influences on the selection of carrion-breeding fauna in Northern Italy and consequently on inferred forensic data (post-mortem intervals and post-mortem transfer), we present details of six indoor–outdoor cases. Results show that the most abundant species was Lucilia sericata, together with other fly species of entomo-forensic interest, belonging to the Calliphoridae and Sarcophagidae families. In particular, for the first time in Italy, we report finding Phormia regina, Lucilia ampullacea, Lucilia caesar and Sarcophaga (Pandelleana) protuberans on fresh cadavers. The active period of L. sericata in Northern Italy, according to previous findings in Southern Europe, revealing clearcut differences with phenologies in Northern Europe, has important consequences in estimating the period (season, months) of death in cases of long post-mortem intervals (several months or years) if empty puparia of this fly are found. According to our results, the distribution of L. sericata in areas with urban sprawl, like Northern Italian regions, cannot be used to evaluate post-mortem transfer from an urban area to a rural one. # 2007 Elsevier Ireland Ltd. All rights reserved. Keywords: Forensic entomology; Lucilia sericata; Lucilia ampullacea; Lucilia caesar; Phormia regina; Sarcophaga (Pandelleana) protuberans; PMI estimation; Post-mortem transfer 1. Introduction stay in shelter and do not eat, is reported for several insect species belonging to various orders as Coleoptera [6], Diptera In forensic entomology, necrophagous insects are useful in [7], Heteroptera [8], etc. studying the post-mortem interval (PMI), post-mortem transfer Thus, climatic, geographic and environmental conditions all (the transfer of the cadaver between different locations after influence the fauna in a particular area. Knowledge of death), and presence of drugs or poisons [1,2]. distributions and preferred habitats is useful in investigating Many species may be used to calculate the PMI, according to post-mortem transfer. stage of cadaver decomposition, body exposure (on soil, buried, The result reported in this paper, according to Grassberger in water, etc.), geographic region and season. In temperate and Frank [9], indicate that entomological evidence must be regions, significant differences between cold and warm seasons evaluated on a regional scale. In particular, no global in necrophagous fauna associated with human remains have generalisations may be made in estimating time of death been reported [3]. Insect phenology and activity (flight, mating precisely. Until now, in evaluating entomological evidence, the and feeding) are temperature-dependent. Cold is not only an paucity of entomo-forensic data for South European regions has obstacle for insect life but, for several species, warmth may obliged researchers to use entomo-forensic data recorded in even be a barrier [4,5]. For example, aestivation, when animals other geographical and biogeographical areas with different fauna and/or environmental conditions, such as North and Central Europe, Southern Africa, Australia and America. * Corresponding author. Tel.: +39 049 827 6309; fax: +39 049 827 6300. To contribute to the interpretation of entomological data in E-mail address: [email protected] (S. Vanin). Northern Italy, we analysed six cases from the entomologic and 0379-0738/$ – see front matter # 2007 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.forsciint.2007.10.006 38 S. Vanin et al. / Forensic Science International 177 (2008) 37–41 medico-legal points of view. Six species belonging to the 2.1.4. Entomo-forensic data Calliphoridae family [Lucilia ampullacea (Villeneuve, 1922); Larvae were collected both from the death scene and during autopsy. Three insect samples, one from the floor near the cadaver, one from the body itself, and Lucilia caesar (Linnaeus, 1758); Lucilia sericata (Meigen, a third, collected during autopsy, were studied. The floor sample included two 1826); Calliphora vicina (Robineau-Desvoidy, 1830); Calli- pupae of Sarcophagidae, one pupa of Calliphoridae, and larvae LIII of the same phora vomitoria (Linnaeus, 1758); Phormia regina (Meigen, families. The Sarcophagidae pupae enclosed two adults, identified as S. 1826)] and one species of Sarcophagidae, Sarcophaga protuberans. One adult of P. regina emerged from the Calliphoridae pupa. (Pandelleana) protuberans (Pandelle´, 1896), were collected. Identification of both larvae and adults after rearing revealed the presence of other S. protuberans and L. sericata specimens. In particular, we focused on the distribution and phenology The samples obtained from the body after discovery and during autopsy of species belonging to the genus Lucilia: these flies, often comprised several larvae (LII and LIII-peakfeeding and postfeeding) of L. involved in entomo-forensic cases in Southern Europe, show sericata and larvae LIII of S. protuberans. No coleopterans or other insects were clearcut differences when compared with the populations of found on the body, in the room where it was discovered, or in the other rooms of Central and Northern Europe [10]. the apartment. In view of the elevated temperature (mean 29 8C) before the discovery and the effect of the indoor environment, it was estimated that P. regina needed 4–6 days to reach the pupal stage [13,14]. Measurement of the L. 2. Materials and methods sericata larvae were compared with available data [15] and revealed the larvae to be 4 days old. No data are currently available for S. protuberans. Therefore, Insects (eggs, larvae and adults) were collected from various body parts, considering the development times calculated for the two Calliphoridae species both at the moment of finding and during autopsy, and from the environment in and adding 1 day for egg laying and hatching, it was estimated that the minimum which the bodies were discovered. Entomological studies were carried out at the PMI was about 5 days. Laboratory of Forensic Entomology, Department of Biology (University of Padova), following the standards and guidelines proposed by the European Association of Forensic Entomology [11]. Some immature specimens were 2.2. Case 2 killed in hot water and stored in 70% alcohol, and some were reared in the laboratory to identify them at the adult stage. Larvae were kept in small cages in 2.2.1. Circumstantial data a thermostatic room (24.0 Æ 1.0 8C) and fed on rotten minced meat. For During the same week as case 1, in the same part of the city, the body of a 66- breeding, days were set at a photoperiod of 14 h of light and 10 h of dark year-old man, wearing a pair of shorts and a vest, was found in his house. He was (D14:L10). lying on the floor on his left side, over a large spot of liquid in advanced Climatic data were obtained from meteorological stations located at not putrefaction. In the room where the body was found, the windows were open. more than 3 km from the discovery site and from measurements performed at that site. A portable Handheld Infrared Thermometer (IMPAC Electronic Gmbh 2.2.2. Autopsy findings Infratherm IN 15 plus) was used to obtain both the environmental (air and soil) On inspection 1 day later, no signs of injury were found, and many larvae and the body (hands, feet and abdomen) temperatures. The measured tempera- were collected from mouth, nose cavities, axillae, chest and clothing. tures were compared by the data obtained by the meteorological stations and, if necessary, recorded during the days after the body discovery. Except in a case 2.2.3. Climatic data (see case 5), no statistical significant differences were found. The same data used in case 1 were examined. All reported values are presented as means Æ S.D. Tests among means were conducted with Student’s t-test. Statistical analyses were carried out with Origin 2.2.4. Entomo-forensic data 7.0 SRO (OriginLab Corporation, Northampton, MA, USA). Zoological Three maggots collected during autopsy from hips, back and clothing were nomenclature follows Minelli et al. [12]. studied. All samples were larvae at the second (LII) and third (LIII) instars of L. sericata, and no other species were found. The minimum PMI was calculated, in 2.1. Case 1 the same way as for case 1, as 48 h. 2.3. Case 3 2.1.1. Circumstantial data At the end of July 2006, the body of an 84-year-old woman was found in her house in the city of Padova. She was lying on the floor, supine, inside a room 2.3.1. Circumstantial data with the windows open. The corpse, wearing pyjamas, a t-shirt and a woollen In early September 2006, a hanged man, 33 years old, was found in a hilly vest, was partially covered by a woollen blanket. area 20 km from the city of Padova. The death, a suicide, had occurred in a small cherry orchard, at the edge of a deciduous wood. The corpse was 40 cm from the ground. No sign of injury was found. 2.1.2. Autopsy findings At external examination the body showed signs of putrefaction, consisting 2.3.2.