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Diptera: Milichiidae) from a Human Cadaver in the Mediterranean Area

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Diptera: Milichiidae) from a Human Cadaver in the Mediterranean Area Forensic Sciences Research ISSN: 2096-1790 (Print) 2471-1411 (Online) Journal homepage: https://www.tandfonline.com/loi/tfsr20 Record of Leptometopa latipes (Diptera: Milichiidae) from a human cadaver in the Mediterranean area Giorgia Giordani, Fabiola Tuccia, Silvia Zoppis, Carla Vecchiotti & Stefano Vanin To cite this article: Giorgia Giordani, Fabiola Tuccia, Silvia Zoppis, Carla Vecchiotti & Stefano Vanin (2019) Record of Leptometopalatipes (Diptera: Milichiidae) from a human cadaver in the Mediterranean area, Forensic Sciences Research, 4:4, 341-347, DOI: 10.1080/20961790.2018.1490473 To link to this article: https://doi.org/10.1080/20961790.2018.1490473 © 2018 The Author(s). Published by Taylor & View supplementary material Francis Group on behalf of the Academy of Forensic Science. Published online: 09 Oct 2018. Submit your article to this journal Article views: 504 View related articles View Crossmark data Citing articles: 1 View citing articles Full Terms & Conditions of access and use can be found at https://www.tandfonline.com/action/journalInformation?journalCode=tfsr20 FORENSIC SCIENCES RESEARCH 2019, VOL. 4, NO. 4, 341–347 https://doi.org/10.1080/20961790.2018.1490473 ORIGINAL ARTICLE Record of Leptometopa latipes (Diptera: Milichiidae) from a human cadaver in the Mediterranean area Giorgia Giordania, Fabiola Tucciaa, Silvia Zoppisb, Carla Vecchiottib and Stefano Vanina aSchool of Applied Sciences, University of Huddersfield, Huddersfield, UK; bDipartimento di Scienze Anatomiche Istologiche Medico Legali e dell’Apparato Locomotore, Section of Legal Medicine, Laboratory of Forensic Genetics, University of Rome "Sapienza", Rome, Italy ABSTRACT ARTICLE HISTORY In forensic entomology, insects are used mainly to obtain information about the time since Received 4 April 2018 death. This information is obtained studying the developmental rate of the first colonizers, Accepted 15 June 2018 principally species in the families Calliphoridae, Sarcophagidae, Muscidae, Stratiomyidae and KEYWORDS Phoridae. However, species belonging to other families can provide information about body Forensic entomology; transfer or the season of the death. Among them Milichiidae are flies rarely reported from postmortem interval; human cases despite the larvae of some species are known as saprophagous feeding on decomposition; plant and animal decomposing matter. A potential cause of the lack of records of these spe- Leptometopa latipes cies from forensic cases can be related with the paucity of descriptions and illustrations of the immature stages. In this article, the entomological samples collected from a human body found inside an apartment in a Maghreb country, in Northern Africa, is reported and Leptometopa latipes (Diptera: Milichiidae) is described in detail. Molecular analysis is also reported to confirm the morphological analysis. Introduction Stratiomyidae, Fanniidae, Piophilidae, Phoridae families are often sampled from the body whereas Forensic entomology is a branch of forensic science species in the families of Syrphidae, Sphaeroceridae, in which insects are used as evidence in legal inves- tigations relating to humans or wildlife [1]. The Heleomyzidae, Sepsidae are only occasionally col- examination, identification and analysis of insects lected. Other species, in the families of Trichoceridae, associated with human remains, combined with the Psycodidae, Milichiidae, Ulidiidae and Drosophilidae, knowledge of insect biology, distribution, phenology are very rare and their presence depends on very and ecology can provide a further level of detail in specific seasonal, geographical or ecological contexts addition to medical and anthropological data in the in addition to the decomposition stage of the body. reconstruction of events occurring close to the time These late species could provide a very detailed infor- of death. In particular, in forensic entomology, nec- mation about the perimortem events, as, for example, rophagous insects are useful in estimating the mini- body transfer or concealment, despite generally they mum postmortem interval (mPMI), the movement are useless for the mPMI estimation because the lack of the cadaver after death (body transfer), the season of developmental data and their unpredictable arrival of death, the presence of drugs or poisons [2] and, on the body. Moreover, the paucity of records of as reported in the last years, in the identification of these species can be related with the lack of a correct the victim in cases of body removal before the identification because no detailed identifications keys investigators’ arrival [3]. are available for the morphological identification of Flies are typically the first insects to colonize a their immature stages and the incompleteness of dead body. The species of flies involved differ from the molecular information about these “secondary” one location to another, but several studies have species in Genbank or in BOLD [4–6]. indicated that the primary species involved belong to The case here reported concerns the finding on a a relatively small number of families: Calliphoridae, human cadaver in advanced decay of several Sarcophagidae and Muscidae [2]. Later during specimens of Leptometopa latipes (Meigen, 1830), a fly the decomposition other species belonging to the species in the family of Milichiidae. A detailed CONTACT Stefano Vanin [email protected], [email protected] Supplemental data for this article is available online at https://doi.org/10.1080/20961790.2018.1490473. Color versions of one or more of the figures in the article can be found online at www.tandfonline.com/tfsr. ß 2018 The Author(s). Published by Taylor & Francis Group on behalf of the Academy of Forensic Science. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 342 G. GIORDANI ET AL. description of the puparium and the adult of the spe- To further confirm the morphological identification, cies are here presented in order to provide support a molecular DNA extraction from adult flies was per- for the identification of specimens from forensic cases. formed. A 737 bp sequence of the cytochrome c oxi- Molecular analysis is also presented and discussed. dase subunit I (COI) mitochondrial gene was amplified V Milichiidae sometimes called “free loader flies” and sequenced. The online system BLASTn R [12]was from their kleptoparasitic habits are small little black used for species identification based on the percentage acalyptrate flies with larvae generally saprophagous, of identity with those available on the online gene usually developing in decomposing plants and ani- banks. The obtained sequence is deposited on mals but also collected from dung and human GenBank (accession number MH069729). Sequences excrements or organic matters accumulated inside of some Milichiidae species were downloaded from the nest of ants or other social insects [2]. GenBank and BOLD and included in the analysis (Supplementary material Table S1). In order to Case description increase the number of species in the analysis a region of only 533 bp was considered. Sequences were aligned In 2006, the corpse of an Italian young man was with Clustal Omega [13]. A phylogenetic tree was built found inside an apartment in a Maghreb country, in using the Neighbour Joining method on MEGA Northern Africa. According to the first autopsy version 7.0 [14]. A bootstrap of 1 000 replicates was report, the cause of death was intoxication by carbon used for testing the robustness of the phylogenetic monoxide. The corpse was transferred to Italy, per- reconstruction. The tree was visualized with iTOL [15]. forming a second postmortem examination; however, In the phylogenetic reconstructions, sequences of the conditions of the remains made impossible deter- Drosophilidae species were considered as outgroup. mining the cause of death. Furthermore, the relatives asked for new research and analysis to confirm his Results identity. In 2013, the corpse was exhumed. Then, bone samples were collected, performing the kinship A large number of puparia and small flies were col- genetic tests. However, due to the poor conditions of lected over the bone surfaces, identifying two differ- the remains, these results were not successful on ent species: Hydrotaea capensis (Wiedemann, 1818) identity matching. The remains were heavily colon- (Diptera: Muscidae), common species reported from ized by insects: large number of puparia and small decomposed and buried remains, and unexpectedly, flies were collected over the bone surface. L. latipes (Diptera: Milichiidae). The identification Entomological samples were sampled, prepared and of the late species was confirmed as well by molecu- analysed following the standards and guidelines of lar analysis. The local alignment of the sequence the European Association for Forensic Entomology belonging to adult flies from this case was per- V (EAFE) [7] and further analysed in order to obtain formed using the online system BLAST R (https:// information useful for the investigation. blast.ncbi.nlm.nih.gov/Blast.cgi). The output revealed a 99% and 97% identity, respectively, with two Materials and methods sequences coded in Genbank as L. latipes KR671912.1 and KR434028.1, with a 0.0 E value. A phylogenetic All the samples were initially observed using a approach was used to
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