ῌMed. Entomol. Zool. Vol. 61 No. 2 p. 115ῌ119 2010῍

Larvae of the family found in the marrow space of skeletal remains during a forensic autopsy

Masashi K>G>CD@>1), Masahito H>IDHJ<>2), Yuichi C=>

1) Laboratory of Tropical Medicine and Parasitology, Institute of International Education and Research, Dokkyo Medical University, Mibu, Shimotsuga, Tochigi 321ῌ0293, Japan 2) Department of Legal Medicine, School of Medicine, Dokkyo Medical University, Mibu, Shimotsuga, Tochigi 321ῌ0293, Japan.

(Received 12 February 2010; Accepted: 17 March 2010)

Abstract: We found the 3rd instar larvae of the family Piophilidae (Diptera) in skeletal remains during a forensic autopsy in late December in the Tochigi Prefecture, Japan. Of note, all the larvae were found in the marrow space of the right femur, thus indicating that the piophilid larvae are able to penetrate the bones of human corpse. This fact demonstrates the importance of a careful examination of skeletal remains, and is emphasizing that forensic pathologists should also examine the inside of bones during a forensic autopsy, because such procedures may provide valuable entomolog- ical evidence.

Key words: Piophilidae, forensic autopsy, , forensic post-mortem examination, bone

little information reported concerning the ICIGD9J8I>DC associated with human cadavers with a long post-mortem period. We Although dipteran flies are well known therefore herein present a case in which for their role in (Chigusa et al., larvae were detected in the skeletal 2005, 2006; Gupta, 2009), maggot therapy remains. (Fleischmann et al., 2004) and also for fo- rensic entomology (Go#,2000; Hitosugi et C6H: PG:H:CI6I>DC al., 2007). Forensic entomology established on the The skeletal remains were found in a fact that insects collected from a corpse mountain forest in the central part of the sometimes provide useful information for Tochigi Prefecture (139῎ῌ140῎E, 36ῌ37῎ investigations. For instance, if the habitat N), Japan, in late December. The forest of a collected is at variance with the where the remains were found is located area where the corpse was found, then the at a slope facing to the west, and the displacement of the corpse should be con- ground was thinly covered with fallen sidered. In addition, for the cadavers with leaves (Fig. 1). a short post-mortem period, the identific- No marked injuries were found on the ation of detailed species and instars of cadaver except for post-mortem changes. collected larvae may provide entomologi- To estimate the age of the remains, bone cal evidence regarding the minimum time density was macroscopically examined since death, even when medical parame- from the femur. When the head (Caput ters are no longer of any value (Hitosugi et femoris) of the right femur was cut by the al., 2007). However, there has so far been striker, dozens of larvae were observed in 116 Med. Entomol. Zool.

regions. Five species have been reported in Japan (Iwasa, 1998); casei (Linnaeus), Stearibia nigriceps (Meigen), Liopiophila varipes (Meigen), Protopiophila contecta (Walker) and Protopiophila latipes (Meigen). All the species were reported to be distributing in Honshu, and some were found in Hokkaido. The piophilid flies are commonly found in variety of protein rich matter. Piophila casei (Linnaeus), which is the most common member of the family, is known as a serious pest of a variety of products, including meat, fish, cheese and leather. Besides the problems they pose to these industries, the fly larvae are also Fig. 1. Forest site where the remains were responsible for causing intestinal myiasis found. (Peckenscneider et al., 1952) and urinary myiasis (el-Serougi, 1991; Saleh and el Sibae, 1993). However, the Piophilidae often serve as forensically significant in- sects. They are commonly found in foren- sic experiments using animal carcasses or beef bait as a trap (Azwandi and Abu Hassan, 2009; Apichat et al., 2007; Gras- sberger and Franc, 2004; Matuszewski et al., 2008; Oliva, 2001; Tabor et al., 2004; Watson and Carlton, 2003; Wol# et al., 2001). Further, the flies have been reported from human cadavers (Nazni et al., 2008; Saigusa et al., 2003, 2006; Sukontason et al., 2001, 2007). The larvae of the P. casei are found during the ad- Fig. 2. The section of the head of the right vanced decomposition phase of a cadaver, femur. Arrows indicate the larvae of the while the adult flies are typically observed family Piophilidae. beginning several days after death (Byrd and Castner, 2009). the inside of the head (Fig. 2). The larvae In the present case, the skeletal remains actively moved about and often leaped. were found in late December in the central The larvae were entomologically iden- part of Japan, Tochigi Prefecture. It was tified to be the 3rd instar larvae of the thought to be too cold at this time of year family Piophilidae (Diptera). After the au- for the flies to deposit their eggs. Saigusa topsy, the remains were forensically deter- et al. (2006, 2008) reported that no insects mined to be those of an adult male. were found in corpses in November and December except for a very small number D>H8JHH>DC of cases in the Iwate Prefecture (39ῌῌ40ῌ N). Concerning the present case, accord- The flies of the family Piophilidae are ing to the Automated Meteorological Data found worldwide, and most of them are Acquisition System (AMeDAS) operated distributed in the temperate and boreal by the Japan Meteorological Agency at Vol. 61 No. 2 2010 117

the nearest observation site from the place the remains were found, the daily average temperature was around 5῎ in early De- cember. However, the daily maximum temperature was over 10῎ on eight of the first 10 days of December. Toward the middle December, there was one relatively warm day with a daily average tempera- ture of 10.4῎ and a maximum tempera- ture of 15.6῎, and this was 12 days before the larvae were found. After that, the temperature decreased, with daily aver- ages around 0῎ (ῌ2῎) and minimum temperatures under 0῎ during the nine days prior to finding the corpse. The tem- perature of the site where the remains were found may have been lower than these data because the altitude of the site is 150ῌ200 m higher than the AMeDAS observation site. Watanabe et al. (2005) described the es- timated time of death by examining the Fig. 3. A head of a right femur in preparation hibernating fly larvae collected from the for a medical student to study. Numerous remains of an individual in Hokkaido (42῍ vascular foramina are present at the neck of the head. The photo was provided courtesy N). Concerning the development of P. of the Department of Macroscopic Anatomy, casei, Saito et al. (1978) reported the dura- Dokkyo Medical University. tion of egg and larval stages at 18ῌ25῎ to be 3days and 8ῌ10 days, respectively. To perature caused by the feeding activity of our knowledge, however, no previous the larvae should be considered as an in- studies on the development of piophilid fluencing factor underlying the survival flies under cold conditions and during the of the larvae (Aggarwal, 2005). winter hibernating stage have ever been In the present case, the larvae were all reported. found in the marrow space of the bone. Because police information indicated We believe that the larvae may have that the corpse had not been moved from penetrated into inside the bone through another location, we concluded that the holes on the surface, such as a vascular piophilid flies deposited their eggs, which foramen, because the no obvious injury were able to survive until at least the 3rd was found on the right femur. Small in- instar, despite the cold conditions. We sects may be able to enter the bones with- believe it is most likely that the flies out any artificial crack or hole, because deposited the eggs on a day when the numerous vascular foramina with a diam- maximum temperature was over 10῎ be- eter greater than 1mm are present on the cause the oviposition of the order Diptera neck of a head of a femur (Fig. 3). Oliva (flies) is negligible at temperatures below (2001) conducted a study of experimental 10῎ (Aggarwal, 2005). The hatched lar- forensic entomology using beef baits, and vae subsequently must have penetrated found the larvae of Piophila sp. in the into the bone, grew and survived in the marrow cavities of bones. During the same spongy substance of the femur. In addi- studies, the small beetle Carpophilus hemi- tion to the insulating ability of the spongy pterus (Linnaeus) (Nitidulidae) was found substance of the bones, an increase in tem- in the medullar cavities. 118 Med. Entomol. Zool.

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