Print version ISSN 0074-0276
Mem. Inst. Oswaldo Cruz vol.104 no.4 Rio de Janeiro July 2009
S VaninI, +; L CaenazzoII; A ArseniII; G CecchettoIII; C CattaneoIV; M TurchettoI
IDipartimento di Biologia
IIDipartimento di Medicina Ambientale e Sanità Pubblica
IIIUOC Tossicologia Forense ed Antidoping, Azienda Ospedaliera, Università di Padova, via Bassi 58/b, 35131 Padova, Italy
IVLaboratorio di Antropologia e Odontologia Forense, Istituto di Medicina Legale, Università degli Studi di Milano
Knowledge of the carrion-breeding insects present at a local level is important and necessary for defining the post-mortem interval. Climate changes and globalisation are affecting species ranges and population dynamics. In this note, we report the incidence of Chrysomya albiceps (Diptera: Calliphoridae) on dead human bodies and carrion in Northern Italy. These data confirm the spread of this species in the Northern regions. The partial sequencing of a 583-bp region of the cytochrome oxidase subunit 1 gene of an Adriatic population did not reveal any difference compared to the same genomic region in the African and South American populations of this species.
Key words: climate change - forensic entomology - insect dispersion - COI
Flies, especially Calliphoridae, are the first insects to arrive on a corpse after death, and, for forensic entomologists, they play a privileged role in defining the post mortem interval (PMI) (Smith 1986). Within the species belonging to Calliphoridae, different trophic behaviours, as well as changes in food preferences, have been described during development. The calliphorid species Lucilia, Calliphora, Protophormia and Phormia are exclusively saprophagous, whereas the young larvae of the Chrysomya species feed on decomposing organic matter but the second and third stage larvae become predatory on the larvae of other Diptera (Ulyett 1950, Braack & Retief 1986, Faria & Godoy 2001). These feeding habits have direct consequences on the composition of body-breeding fauna (Grassberger et al. 2003).
Chrysomya albiceps (Wiedemann, 1819) (Diptera: Calliphoridae) is a well-known hemisynanthropic fly (Verves 2004). Larvae develop in animal carrion, human cadavers and in faeces and they can cause primary and secondary cutaneous myiasis in mammals (Zumpt 1965, Marchenko 1985, Smith 1986, Hall & Smith 1993, Hall & Wall 1995, Paraluppi & Linhares 1995, Queiroz et al. 1997, Soler 2000, Adham et al. 2001, Madeira 2001, Grassberger et al. 2003). Adults of both sexes feed on faeces, carrion and rotting fruit (Martinez-Sanchez et al. 2000).
C. albiceps is widely distributed in the Southern Pa-laearctic, Northern Oriental and Afrotropical regions and was recently found in Central and South America (Baumgartner & Greenberg 1984, Schumann 1986, Grassberger et al. 2003, Verves 2004, Richards et al. 2007).
In their work on the distribution of C. albiceps in Southern Europe and on the applications and importance of this species in forensic entomology, Grassberger et al. (2003) reported the probable distribution of this species in the Turano-European region (indicated in the text as the Palaearctic distribution). In the Grassberger article, the presence of C. albiceps is indicated in Italy only in the city of Bari (Southern Italy) (Introna et al. 1998) and probably from other locations in the South and Center of Italy and the Southern part of the Po Plain. Furthermore, in the checklist of Italian fauna (Minelli et al. 1993-1995), this species is is only listed for the Southern peninsular regions and for Sicily.
MATERIAL AND METHODS
In the last year, an increasing interest in forensic entomology in Italy and in Europe in general and a call for the re-writing of the carrion-breeding insect list at a local scale (Grassberger & Frank 2004, Vanin et al. 2008) encouraged the collection of new records of "forensically important" species. The continuous sampling of these species is a priority in order to investigate the dispersion of new species caused by climate change and globalisation (Turchetto & Vanin 2004a, b) and to prevent and control sanitary and veterinary emergences. During 2007, three collections of C. albiceps species from human corpses and animal carrion were performed in Northern Italy (Fig. 1). Entomological sampling and studies were performed following the standards and guidelines proposed by the European Association of Forensic Entomology (Amendt et al. 2006).
RESULTS AND DISCUSSION
C. albiceps maggots were first found in the clothes of a young dead woman found indoors at end of July 2007, close to the city of Chioggia (Venice) (45°08'N; 12°15''E). The body was in a stage of advanced decomposition and signs of strangling were evident. The minimal PMI was estimated to be seven-eight days. In this case, larvae (LIII) and pupae of Sarcophaga sp. were also collected. No adults were obtained from the larvae.
In August 2007, a second sample of 11 maggots (LIII) was collected during the autopsy of a 57-year-old woman, found dead in her house in the city of Padua (45°24'N; 11°52'E). The body was in a stage of advanced decomposition and was partially mummified. No sign of injury was found. On the body, larvae (LIII) of C. albiceps and larvae (LIII) and pupae of Phormia regina, Sarcophagidae and Phoridae species were collected. The proper identifications were confirmed by analysis of the adults obtained from the larvae. The temperature of the collection period ranged between 24.9 ± 3.6°C during the day and 20.7 ± 2.1°C during the night (max: 32.9°C; min 15.7°C).
The third sample was obtained during an experiment carried out on fresh and burned pigs in the outskirts of Milan (45°27'N; 9°11'E). Larvae were collected during July and August 2007. Identification of the samples was confirmed by analysis of the adults obtained from the larvae.
In addition, several collections of adults were performed during the summer and fall of 2008 in the Northern and Central Italian regions and on the Island of Krk, Croatia (Fig. 1).
The collections of C. albiceps reported in this note validate the previously doubtful and uncertain records from Northern Italy, which were based on single immature specimens that were destroyed or misplaced after identification. All of our specimens, except for the individuals used for the molecular analyses, are stored in the private collection of SV (Treviso).
It is worth mentioning that the presence of C. albiceps in Northern Italy confirms the spread of this species in the Northern regions (Grassberger et al. 2003, Verves 2004). The dispersion of this species towards the Northern regions as well as its demographic explosion are correlated to the climate change, as has been demonstrated for several species of insects, vertebrates and plants (Turchetto & Vanin 2004a, Maistrello et al. 2006, Parmesan 2006). Neither the studies of the dipterological collections of several Italian Natural History Museums nor the recent research performed (with different collection methods) in the beech-wood belt in the Northern Apennines revealed the presence of this species in the past (Cerretti & Vanin 2003, Raffone 2005a, b, Vanin & Lencioni 2006).
A partial sequence (583 bp) of the cytochrome oxidase subunit 1 gene (COI) (EU503116) was obtained from larvae collected in the city of Chioggia by PCR as reported in Nelson et al. (2007). The sequences were used to confirm the morphological identification of the larvae. The sequence alignment of the available COI sequences of C. albiceps from Africa (AB112836, AB112839, AB112840, AB112842, AB112849, AB112851, AB112854, AB112858, AB112865, AF083657) (Wells & Sperling 1999, Harvey et al. 2003) and South America (Wells & Sperling 1999) (Fig. 2), performed with ClustalW software (Thompson et al. 1994), revealed an identity of 100% and of 97.78%, respectively, with the sequences of C. rufifacies (AF083658). These data confirm the morphological identification and indicate that the sequence we used cannot be used to distinguish populations. Identification of more informative sequences will be necessary in order to understand the origin of the dispersion of these species and to investigate on bodies probably transferred for long distance.
Adham FK, Abdel MA, Tawfik MAA, El Khateeb RM 2001. Seasonal incidence of the carrion breeding blowflies Lucilia sericata (Meigen) and Chrysomya albiceps (Wied.) (Diptera: Calliphoridae) in Abu-Rawash Farm, Giza, Egypt. Assiut Vet Med J 49: 377-383. [ Links ]
Amendt J, Campobasso CP, Gaudry E, Reiter C, LeBlanc HN, Hall MJR 2006. Best practice in forensic entomology-standards and guidelines. Int J Legal Med 121: 90-104. [ Links ]
Baumgartner DL, Greenberg B 1984. The genus Chrysomya (Diptera: Calliphoridae) in the New World. J Med Entomol 21: 105-113. [ Links ]
Braack LEO, Retief PF 1986. Dispersal, density and habitat preference of the blow-flies Chrysomyia albiceps (WD.) and Chrysomyia marginalis (WD.) (Diptera: Calliphoridae). Onderstepoort J Vet Res 53: 13-18. [ Links ]
Cerretti P, Vanin S 2003. Calliphoridae. In P Cerretti, A Tagliapietra, M Tisato, S Vanin, F Mason, M Zapparoli, Artropodi dell'orizzonte del faggio nell'Appennino Settentrionale, Primo contributo, Conservazione Habitat Invertebrati 2, Gianluigi Arcari Editore, Mantova, p. 184-186. [ Links ]
Faria LDB, Godoy WAC 2001. Prey choice by facultative predator larvae of Chrysomya albiceps (Diptera: Calliphoridae) Mem Inst Oswaldo Cruz 96: 875-878. [ Links ]
Grassberger M, Frank C 2004. Initial study of arthropod succession on pig carrion in a Central European urban habitat. J Med Entomol 41: 511-523. [ Links ]
Grassberger M, Friedrich E, Reiter C 2003. The blowfly Chrysomya albiceps (Wiedemann) (Diptera: Calliphoridae) as a new forensic indicator in central Europe. Int J Legal Med 117: 75-81. [ Links ]
Hall MJR, Smith KGV 1993. Diptera causing myiasis in man. In RP Lane, RW Crosskey, Medical Insects and Arachnids, Chapman & Hall, London, p. 429-469. [ Links ]
Hall MJR, Wall R 1995. Myiasis of humans and domestic animals. Adv Parasitol 35: 257-334. [ Links ]
Harvey ML, Mansell MW, Villet MH, Dadour IR 2003. Molecular identification of some forensically important blowflies of Southern Africa and Australia. Med Vet Entomol 17: 363-369. [ Links ]
Introna F, Campobasso CP, Di-Fazio A 1998. Three case studies in forensic entomology from southern Italy. J Forensic Sci 43: 210-214. [ Links ]
Madeira NG 2001. Would Chrysomya albiceps (Diptera: Calliphoridae) be a beneficial species? Arq Bras Med Vet Zoo 53: 157-161. [ Links ]
Maistrello L, Lombroso L, Pedroni E, Reggiani A, Vanin S 2006. Summer raids of Arocatus melanocephalus (Heteroptera, Lygaeidae) in urban buildings in Northern Italy: is climate change to blame? J Therm Biol 31: 594-598. [ Links ]
Marchenko MI 1985. Characters of the development of fly Chrysomya albiceps Wd. (Diptera: Calliphoridae). Entomol Obozr 64: 79-84. [ Links ]
Martinez-Sanchez A, Rojo S, Marcos-Garsia MA 2000. Annual and spatial activity of dung flies and carrion in a Mediterranean holm-oak pasture ecosystem. Med Vet Entomol 14: 56-63. [ Links ]
Nelson LA, Wallman JF, Dowton M 2007. Using COI barcodes to identify forensically and medically important blowflies. Med Vet Entomol 21: 44-52. [ Links ]
Paraluppi ND, Linhares AX 1995. Calliphoridae (Diptera) em Manaus: 3. Atratividade seletiva das iscas em relação às fases do desenvolvimento ovariano em três espécies de Chrysomya Robineau-Desvoidy. Rev Bras Entomol 39: 121-124. [ Links ]
Parmesan C 2006. Ecological and evolutionary responses to recent climate change. Annu Rev Ecol Evol Syst 37: 637-669. [ Links ]
Queiroz MMC, de Mello RPD, Lima MM 1997. Morphological aspects of the larval instars of Chrysomya albiceps (Diptera: Calliphoridae) reared in the laboratory. Mem Inst Oswaldo Cruz 92: 187-196. [ Links ]
Raffone G 2005a. Catalogo dei ditteri della Laguna di Venezia. II. Acroceridae, Opomyzidae, Calliphoridae, Sarcophagidae (Insecta, Diptera Brachycera). Boll Mus Civ St Nat Venezia 56: 103-111. [ Links ]
Raffone G 2005b. Osservazioni faunistiche sui Calliphoridae italiani. Boll Soc Entomol Ital 138: 149-55. [ Links ]
Richards CS, Paterson ID, Villet MH 2007. Estimating the age of immature Chrysomya albiceps (Diptera: Calliphoridae), correcting for temperature and geographical latitude. Int J Legal Med 122: 271-279. [ Links ]
Schumann H 1986. Family Calliphoridae. In L Papp, A Soos, Catalogue of Palaearctic Diptera 12. Calliphoridae - Sarcophagidae, Academy Press, Budapest, p. 11-58. [ Links ]
Smith KGV 1986. A Manual of Forensic Entomology, Cornell Univ. Press, London, British Museum of Natural History, 205 pp. [ Links ]
Soler CMD 2000. El estudio de las miasis en Espana duranto los ultimos cien anos. Ars Pharm 41: 19-26. [ Links ]
Thompson JD, Higgins DG, Gibson TJ 1994. CLUSTALW: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22: 4673-4680. [ Links ]
Turchetto M, Vanin S 2004a. Forensic entomology and climatic change. Forensic Sci Int 146 (Suppl.): 207-209. [ Links ]
Turchetto M, Vanin S 2004b. Forensic entomology and globalisation. Parassitologia 46: 187-90. [ Links ]
Ulyett GC 1950. Competition for food and allied phenomena in sheep-blowfly populations. Philos Trans R Soc Lond B Biol Sci 234: 77-174. [ Links ]
Vanin S, Lencioni V 2006. La collezione ditterologica del Museo Tridentino di Scienze Naturale (Trento, Italia). Studi Trent Sci Nat Acta Biol 82: 55-59. [ Links ]
Vanin S, Tasinato P, Ducolin G, Terranova C, Zancaner S, Montisci M, Ferrara P, Turchetto M 2008. Use of Lucilia species (Diptera: Calliphoridae) for forensic investigations in Southern Europe. Forensic Sci Int 177: 37-41. [ Links ]
Verves YuG 2004. Records of Chrysomya albiceps in the Ukraine. Med Vet Entomol 18: 308-310. [ Links ]
Wells JD, Sperling FA 1999. Molecular phylogeny of Chrysomya albiceps and C. rufifacies (Diptera: Calliphoridae). J Med Entomol 36: 222-226. [ Links ]
Zumpt F 1965. Myasis in man and animals in the Old World, Butterworths, London, 267 pp. [ Links ]
Received 12 August 2008
Accepted 3 March 2009