Kom Sukontason

Forensic entomology cases in Thailand: a review of cases from 2000 to 2006

This paper presents and discusses 30 cases of cadavers that had been transferred for forensic entomology investigations to the Department of Forensic Medicine, Faculty of Medicine, Chiang Mai University, northern Thailand, from 2000 to 2006. Variable death scenes were determined, including forested area and suburban and urban outdoor and indoor environments. The fly specimens found in the corpses obtained were the most commonly of the blow fly of family Calliphoridae, and consisted of Chrysomya megacephala (F.), Chrysomya rufifacies (Macquart) Chrysomya villeneuvi Patton, Chrysomya nigripes Aubertin, Chrysomya bezziana Villeneuve, Chrysomya chani Kurahashi, Lucilia cuprina (Wiedemann), Hemipyrellia ligurriens (Wiedemann), and two unknown species. Flies of the family Muscidae [Hydrotaea spinigera Stein, Synthesiomyia nudiseta (Wulp)], Piophilidae [Piophila casei (L.)], Phoridae [Megaselia scalaris (Loew)], Sarcophagidae [Parasarcophaga ruficornis (F.) and three unknown species], and Stratiomyiidae (Sargus sp.) were also collected from these human remains. Larvae and adults of the beetle, Dermestes maculatus DeGeer (Coleoptera: Dermestidae), were also found in some cases. Chrysomya megacephala and C. rufifacies were the most common species found in the ecologically varied death scene habitats associated with both urban and forested areas, while C. nigripes was commonly discovered in forested places. S. nudiseta was collected only from corpses found in an indoor death scene.

Ultrastructure of adhesive device in fly in families calliphoridae, muscidae and sarcophagidae, and their implication as mechanical carriers of pathogens.

The ultrastructure of adhesive device or the pulvilli, pad-like structure between the tarsal claws of the legs, is presented in the blowfly (Calliphoridae), housefly and relatives (Muscidae), and flesh fly (Sarcophagidae) through scanning electron microscopy. The blowfly species were Chrysomya chani, Chrysomya nigripes, Chrysomya pinguis, and Chrysomya villeneuvi; while those of the housefly and relatives were Musca domestica and Hydrotaea chalcogaster, respectively. Fresh fly species included Boettcherisca peregrina and Liosarcophaga dux. Numerous tenent setae were observed on the ventral side of the pulvilli. Two features of the tip of the tenent setae were found as a spatula-like (in C. chani, C. pinguis, C. nigripes, M. domestica, H. chalcogaster, B. peregrina, and L. dux) and spoon-like tip (in C. villeneuvi). Transmission electron microscopy of the tenent setae in M. domestica revealed the electron-lucent centrally located, suggesting an adhesive substance. These results provided anatomical information that allow us to not only understand the successful attachment of flies to smooth surfaces but also clarify their role as a mechanical carrier of microorganisms.

First Report of Human Myiasis Caused by Chrysomya megacephala and Chrysomya rufifacies (Diptera: Calliphoridae) in Thailand, and Its Implication in Forensic Entomology

Abstract We report a forensic entomology case associated with human myiasis in Chiang Mai Province, northern Thailand. The remains of a 53-yr-old-male were concurrently infested with third instars of the two blow fly species, Chrysomya megacephala (F.) and Chrysomya rufifacies (Macquart), near a severe tumor lesion presented on the lower right leg. The presence of third instars, ≈5 d old, on the day following postmortem indicated that myiasis occurred before death. This is the first report of both fly species acting as a myiasis-producing agent in Thailand. Unsynchronized data between the age of fly larvae due to myiasis premortem and verified age/condition of the corpse suggest a potential complication and error in the estimation of postmortem interval if other predisposition fly infestations are not considered.

Effects of eucalyptol on house fly (Diptera: Muscidae) and blow fly (Diptera: Calliphoridae).

The effects of eucalyptol were evaluated against the house fly, Musca domestica L., and blow fly, Chrysomya megacephala (F.). The bioassay of adults, using topical application, indicated that M. domestica males were more susceptible than females, with the LD50 being 118 and 177 microg/fly, respectively. A higher LD50 of C. megacephala was obtained; 197 microg/fly for males and 221 microg/fly for females. Living flies of both species yielded a shorter life span after being treated with eucalyptol. The bioassay of larvae, using the dipping method on the third instar, showed that M. domestica was more susceptible than C. megacephala, with their LC50 being 101 and 642 microg/microl, respectively. The emergence of adults, which had been treated with eucalyptol in larvae, decreased only in M. domestica. Having the volatile property, fumigation or impregnated paper test of eucalyptol or the efficacy of repellence or attractiveness merits further investigations to enhance bio-insecticidal efficacy.

The First Documented Forensic Entomology Case in Thailand

Abstract The forensic entomology case described herein is the first such case documented in Thailand. A mummified corpse of a 32-yr-old man was discovered in a forested habitat, with the larvae of six species of flies (Diptera) found in association with the corpse at the time of its discovery, i.e., those of Hydrotaea (=Ophyra) spinigera Stein (family Muscidae), Piophila casei (L.) (family Piophilidae), Megaselia scalaris (Loew) (family Phoridae), Sargus sp. (family Stratiomyidae), and larvae of two unidentified flesh fly species (family Sarcophagidae). The presence and age of the larval specimens of P. casei, M. scalaris, and H. spinigera gave entomological evidence that the postmortem interval for the corpse was 3–6 mo. This report also documents some of the forensically important fly species that occur in Thailand.

Morphology of immature stages of Hemipyrellia ligurriens (Wiedemann) (Diptera: Calliphoridae) for use in forensic entomology applications

In forensic investigations, all immature stages of flies (egg, larvae, and puparium) can serve as entomological evidence at death scenes. These insects are primarily used to estimate the post mortem interval (PMI), but can also be involved in the analysis of toxic substances, determining manner of death, and in indicating relocation of a corpse in homicide cases. In this study, we present the morphology of the egg, larvae, and puparium of Hemipyrellia ligurriens, a blow fly species of forensic importance in Thailand. Examination was conducted using scanning electron microscopy (SEM). The egg stage was found to display a relatively wide plastron region (or median hatch line area) that spans almost the entire length of the egg. The median hatch line is oriented in an upright position. External chorionic sculpture of the egg is present in a hexagonal pattern whose reticular boundaries are slightly elevated. In the larval stages, the most prominent morphological changes were detected upon comparison of the first to the second instar; whereas, the differences between second and third instar larvae were less obvious outside of the increase in number of posterior spiracular slits. Most of the major differences involve body size and structure of the anterior and posterior spiracles. Each anterior spiracle in both the second and third instars projects five to seven papillae apically. Each posterior spiracular disc of a third instar exhibits a complete peritreme, three spiracular slits, and a prominent button that is ventromedially located. The puparium is coarctate and features a clustered bubble membrane comprised of ≈57 mammillate structures positioned dorsolaterally on each side of the first abdominal segment in young puparia. This feature is replaced by short, tubular respiratory horns in aged puparia. This study provides more detailed exposure of important morphological features that can be used for accurate identification of immature stages of H. ligurriens. Information presented can aid in forensic investigations involving this fly species.

Differentiation of the Third Instar of Forensically Important Fly Species in Thailand

Abstract Differentiation of the third instar of forensically important fly species in Thailand was performed using light microscopy, based on their morphological criteria for fly identification. Four species of the family Calliphoridae [Chrysomya rufifacies (Macquart), Chrysomya megacephala (F.), Chrysomya nigripes Aubertin, and Lucilia cuprina (Wiedemann)] and two species of the family Muscidae [Musca domestica L. and Hydrotaea (=Ophyra) spinigera Stein] were examined in this study, with the features of the anterior spiracles, dorsal spines between the prothorax and mesothorax, and posterior spiracles being emphasized. The comparisons, presented herein, should be helpful for forensic practitioners to readily distinguish the third instars of fly species found associated with human cadavers, before their use for further forensic investigations.

Clonorchis sinensis: glutathione S-transferase as a serodiagnostic antigen for detecting IgG and IgE antibodies

Human Clonorchis sinensis infection is endemic in East Asian countries. Glutathione S-transferases (GSTs) are anti-oxidant enzymes found in all living creatures as well as in trematodes. In this study, we examined the recombinant 26kDa GST protein of C. sinensis (Cs26GST) for its serodiagnostic antigenicity toward IgG and IgE antibodies by ELISA and immuno-enhanced chemiluminescence, respectively. In IgG ELISA, recombinant Cs26GST showed 33.3% sensitivity and 100% specificity for trematode-infected human sera. In the case of the IgE antibody, recombinant Cs26GST showed 50.0% sensitivity and 93.2% specificity for clonorchiasis infection. We propose that the recombinant Cs26GST is a potent serodiagnostic antigen for detecting C. sinensis-specific IgG and IgE antibodies, and that it be best used as an antigenic cocktail in combination with other antigens.

Surface ultrastructure of Chrysomya rufifacies (Macquart) larvae (Diptera: Calliphoridae).

Abstract The surface ultrastructure of all larval instars of Chrysomya rufifacies (Macquart) is described by means of scanning electron microscopy (SEM). Morphological changes were greatest from the first to the second instar, but less from the second to the third instar. Most of these changes involved the structure of the anterior spiracle, posterior spiracle, integument of the body, and mouthhooks. Modification of the mouthhooks, especially in the third instar, are helpful in explaining the ferocious feeding ability of the older maggots. The common name of “hairy-maggot” for C. rufifacies is only appropriate for the second and third instars because of their elongated tubercles along the body, whereas this name is not descriptive of the first instar that lack tubercles.

Morphology of the puparia of the housefly, Musca domestica (Diptera: Muscidae) and blowfly, Chrysomya megacephala (Diptera: Calliphoridae)

Examination of the puparia of the housefly, Musca domestica L. and blowfly Chrysomya megacephala (F.), through scanning electron microscopy (SEM), revealed many differences in the profile of their morphology. Special attention was focused on puparial characteristics used to differentiate between the two fly species studied, and between other forensically important flies. Results of this study indicate that the housefly puparia are almost evenly rounded at both ends and the anterior spiracle bears six papillae. A pair of pupal respiratory horns is found laterally before the posterior boundary of the first abdominal segment, bearing numerous papillae that have a longitudinal opening along the oval convex base. The peritreme of each posterior spiracle forms a crude forward or reverse D-shape, encircling three sinuous slits. The blowfly pupariums anterior spiracle contains 8–12 papillae. The pupal respiratory horns protrude slightly and in some specimens a group of ~38 globules on the bubble-like membrane may be observed. Each of the posterior spiracles is more or less an oval- shaped peritreme, encircling three straight spiracular slits. The anatomical features presented herein allow for the differentiation of puparia of the two fly species studied and could prove useful in future forensic entomological assessments.