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Forensically Important Calliphoridae (Diptera) Associated with Animal and Human Decomposition in the Czech Republic: Preliminary Results

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Forensically Important Calliphoridae (Diptera) Associated with Animal and Human Decomposition in the Czech Republic: Preliminary Results ISSN 1211-3026 Čas. Slez. Muz. Opava (A), 62: 255-266, 2013 DOI: 10.2478/cszma-2013-0024 Forensically important Calliphoridae (Diptera) associated with animal and human decomposition in the Czech Republic: preliminary results Hana Šuláková & Miroslav Barták Forensically important Calliphoridae (Diptera) associated with animal and human decomposition in the Czech Republic: preliminary results. – Čas. Slez. Muz. Opava (A), 62: 255-266, 2013. Abstract: Two studies to establish standards of sampling of entomological evidence for crime scene technicians and forensic experts in the Czech Republic were pursued in years 2011 to 2013. During experiments, pigs (Sus scrofa f. domestica Linnaeus, 1758) were used as models for human bodies and important data about succession of decomposition of large carcasses were also obtained. Altogether 21 species of Calliphoridae were collected, of which ten are classified as forensically important: Lucilia caesar (Linnaeus, 1758), Calliphora vicina Robineau-Desvoidy, 1830, Phormia regina (Meigen, 1826), Protophormia terraenovae (Robineau-Desvoidy, 1830), Lucilia illustris (Meigen, 1826), Lucilia ampullacea Villeneuve, 1922, Calliphora vomitoria (Linnaeus, 1758), Lucilia sericata (Meigen, 1826), Lucilia silvarum (Meigen, 1826), and Cynomya mortuorum (Linnaeus, 1761). The next eleven species belonged to genera Bellardia Robineau-Desvoidy, 1836, Melinda Robineau- Desvoidy, 1830, Onesia Robineau-Desvoidy, 1830, and Pollenia Robineau-Desvoidy, 1830; the blow flies associated with earthworms and snails, and their role in decomposition is discussed. Finally, our results stated that a pyramidal trap along with pitfall traps and rearing of larvae from the carcass are suitable tool for successional studies of forensically important invertebrates. Key words: Diptera, Calliphoridae, decomposition, forensic entomology, pyramidal trap Introduction Forensic entomology is a particular field of criminalistics which is based on knowledge of invertebrate fauna succession on cadavers. The main aim of the forensic discipline is to establish time of death for a human corpse. The methods of forensic entomology are usually the most accurate after 72 hours of decomposition (Daněk 1990) and among the carrion- feeding species the family Calliphoridae, or blow flies, includes genera of greatest importance to the forensic entomology and provides the most accurate results (Greenberg 1991, Smith 1986). Blow flies are medium-sized to large robust flies with a body length from about 4.0 to 16.0 mm. Body colour variable, but most Central European species are either black or metallic green to blue, often with a silvery to yellowish microtomentum. A great number of the calliphorids develop in decaying organic matter, including carrions as well as human dead bodies (Doskočil 1977, Erzinçlioğlu 1996, Smith 1986). Some of them are important from a hygienic point-of-view since adult blow flies visit faeces, fresh or cooked meat, fish, dairy products, and wounds. It makes many species potential vectors of bacteria, viruses, protozoans and helminths. Several species are synanthropic (Nuorteva 1963) and a few Palaearctic species are involved in myiasis (Zumpt 1965). Altogether 115 species of the family Calliphoridae are known to occur in Europe (Rognes 2013), of which 58 are listed in the Checklist of Diptera of the Czech Republic and Slovakia from the Czech Republic (47 in Bohemia, 57 in Moravia) (Kubík & Országh 2009). Two experiments focused on upright techniques and standards of sampling of entomological traces at a crime scene were pursued in Prague from 2011 to 2013. Two carcasses of pigs (Sus scrofa f. domestica Linnaeus, 1758) were used to imitate the real crime scenes with human corpses. The main aim of the tests was to establish standards and guidelines of collection, preservation, packaging and transport of entomological samples 255 mainly for crime scene technicians and forensic experts in the Czech Republic with reference to quality assurance (Amendt et al. 2007). However data about succession of decomposition of a large carcass, similar to a human corpse, and activity of insect and other arthropods were also available. Studies of the arthropod fauna associated with carrions are important wherever forensic entomology is used because the fauna vary with specific geographic parameters of each region. Both experiments provided interesting data witch upgraded a basic database of the forensically important arthropods in the Czech Republic. In this paper, we are presenting the first results of forensically important members of family Calliphoridae associated with animal and human decomposition in conditions of the Czech Republic which were obtained from both trials. We are also referring to interesting and important factors which occurred or could influence colonization of both cadavers by blow flies and results of testing a pyramidal trap as a collecting method used in forensic field studies with large animals. Material and methods Description of localities The locality of the first experiment, labelled as “Hrdlořezy 2011-12” was situated in a fenced ground of the Police school in Prague - Hrdlořezy, the eastern suburb of the capital. The experimental site consisted of bushy area with smaller opening and deciduous trees in the vicinity. The site was in an open, sunlit place with light, sandy soil covered by ruderal plants and grass (Fig. 1). Geographic coordinates: 50°5’22" N, 14°30’19" E; altitude: 240 m amsl. The second experiment labelled as “Troja 2012-13” was conducted in a fenced experimental fields of the Czech University of Life Sciences in Prague, in district Troja, the northern part of the capital. The area was on west-facing slope near Vltava River, in a flooding zone. The site was in a grass-covered, open, sunlit place with fruit tree species and bushes in vicinity. Geographic coordinates: 50°7'16" N, 14°23'53" E; altitude: 185 m amsl. Both research sites were chosen as typical locations of founding dead human bodies in urbanized areas in the Czech Republic. Description of experiments and collecting methods Pig carcasses were used as models for human decomposition. In the experiment “Hrdlořezy 2011-12”, a pig, at about 65 kg, was killed on 13 July 2011 by a single shot to the front of its head with a 0,22 caliber rifle and moved to the research site within 20 minutes. The carcass was foil-wrapped during its transport to avoid an earlier oviposition before its exposition on the site. On the field, a temperature probe was inserted into centre of the carcass via a small thoracic entrance wound to measure inside temperature afterwards the pig was dressed in a cotton shirt and overalls to imitate a common homicide crime scene (Fig. 2). The date of placing the pig on the site, 13 July 2011, was designed as day 1. The experiment was conducted from 13 July 2011 (day 1) to 18 October 2012 (day 464) until the carcass decomposition was finished. Adult flies were collected over the pig with insect net during control visits and by pitfall traps filled with salt solution and detergent up to two thirds. Control intervals were: day 1 to 17 once a day; day 17 to 62 once two to three days, day 62 to 195 once ten to fourteen days and day 195 to 464 once a month. The second pig, at about 53 kg, used in the experiment “Troja 2012-13”, died of natural causes (no bleeding wound) in the evening on 19 March 2012 and was stored in a coolroom (at about 6 °C) to avoid an earlier oviposition. On 20 March 2012, the carcass was transported on the experimental field, dressed in T-shirt and trousers to imitate death of a person from disease or undercooling, the usual causes of death for displaced persons. The experiment was conducted from 20 March 2012 (day 1) to 6 June 2013 (day 444) until a flood in Prague damaged the experimental site. Adult flies were collected over the pig with pyramidal trap (described by Barták & Roháček 2011) which was placed above the carcass for all time duration of the trial. The trap was situated 20 to 40 cm above ground or vegetation level to make accessible the carcass to insect, base of the trap was 2x2 m in size (Fig. 3). Head of the trap was filled with mixture: 1.5 l of water, 2 ml of 36-38% formaldehyde and 1 ml of detergent. Captured material was collected: day 1 to 197 once a week, day 197 to 267 every fourteen days, day 267 to 393 once a month and then until the end of the trial on day 444 again every fourteen days. The intervals corresponded seasons. Egg and larval samples were taken from the pigs during each control visit. Eggs and larvae were placed in plastic containers with pork meat for larval food and sand substrate for pupation. They were reared to the adult stage to ascertain species which used the carcasses for their development. 256 Figs 1-4: 1 - Experimental site in Prague – Hrdlořezy used during the first trial. 2 - Dressed pig with bleeding wound on its head used as a human model during Hrdlořezy trial. 3 - Experimental pig beneath the pyramidal trap on Troja site. 4 - “Hrdlořezy 2011-12” experiment: adults of Lucilia caesar on the bleeding wound ten minutes after exposition of the killed pig. (All photos by Hana Šuláková) 0,20% 2,06% Calliphora vicina 2,16% 5,70% 1,97% 2,26% Calliphora vomitoria 5,60% Protophormia terraenovae 1,57% Phormia regina Lucilia sericata Lucilia caesar Lucilia illustris Lucilia ampullacea 78,47% Cynomya mortuorum "Hrdlořezy 2011-12" Fig 5: Percentage abundance of the forensically important blow flies collected during “Hrdlořezy 2011-12” experiment. 257 Species 1/VII 2/VII 1/VIII 2/VIII 1/IX 2/IX 1/X 2/X XI XII I II III IV V VI VII VIII IX X 2011 2011 2011 2011 2011 2011 2011 2011 2011 2011 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 Calliphora vicina Calliphora vomitoria Protophormia terraenovae Phormia regina Lucilia sericata Lucilia caesar Lucilia illustris Lucilia ampullacea Cynomya mortuorum Legend: 0-5 5-20 21-100 ≥ 101 Fig 6: Successive abundance of forensically important Calliphoridae collected during “Hrdlořezy 2011-12” experiment.
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