Nophawan Bunchu

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.

Fine Structure of the Eggshell of the Blow Fly, Lucilia cuprina

Abstract The fine structure of the eggshell of blow fly, Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae), was examined using scanning and transmission electron microscopy. Eggs, 1.09±0.07 mm in length and 0.25±0.05 mm in width, bore a relatively wide plastron that extending along almost the entire length. The polygonal pattern of chorionic sculpture was indistinct. The ultrathin section indicated a multi-layered eggshell having an exochorion, outer endochorion, pillars, an inner endochorion, innermost chorionic layer, and a wax layer. This study provides new information about the fine morphology of blow flies eggs. A key to differentiate the eggs of forensically important flies in Thailand is given.

Observations of the third instar larva and puparium of Chrysomya bezziana (Diptera: Calliphoridae)

Observations on the ultrastructure of the third instar larva and puparium of the Old World screw-worm fly, Chrysomya bezziana, are presented utilizing both light microscopy (LM) and scanning electron microscopy (SEM). Results of this study indicate that the shape of the intersegmental spines between the pro- and mesothorax markedly differ from other blow fly species (Chrysomya megacephala, Chrysomya rufifacies, Chrysomya nigripes, Lucilia cuprina) in being broad-based with sharp recurved tips. Other characters such as the posterior spiracles, number of papillae on the anterior spiracles, oral grooves, and posterior spiracular hairs also differ. The strong and robust mouthhooks may explain the ability of larvae to penetrate deeply into human tissues. Perforated sieve plates covered with antler-like projections were observed within the anterior spiracles of the puparium of C. bezziana. The posterior spiracular discs each bear three spiracular slits with ≈2-μm wide openings that were viewed either open or closed by a membrane underneath. This study expands our knowledge of the fine details of the external morphology of both the third instar larva and puparium of C. bezziana, which is an obligatory myiasis-producing species in many regions. A key to differentiate the third instar of C. bezziana from other blow flies in Thailand is given.

Morphology and Developmental Rate of the Blow Fly, Hemipyrellia ligurriens (Diptera: Calliphoridae): Forensic Entomology Applications

Hemipyrellia ligurriens (Diptera: Calliphoridae) is a forensically important blow fly species presented in many countries. In this study, we determined the morphology of all stages and the developmental rate of H. ligurriens reared under natural ambient conditions in Phitsanulok province, northern Thailand. Morphological features of all stages based on observing under a light microscope were described and demonstrated in order to use for identification purpose. Moreover, development time in each stage was given. The developmental time of H. ligurriens to complete metamorphosis; from egg, larva, pupa to adult, took 270.71 h for 1 cycle of development. The results from this study may be useful not only for application in forensic investigation, but also for study in its biology in the future.

Susceptibility to temephos, permethrin and deltamethrin of Aedes aegypti (Diptera: Culicidae) from Muang district, Phitsanulok Province, Thailand

OBJECTIVE To investigate the susceptibility to temephos, permethrin and deltamethrin of Aedes aegypti (Ae. aegypti), collected from areas with high incidence of dengue hemorrhagic fever cases in Phitsanulok Province, Thailand. METHODS The F1 progenies of Ae. aegypti colony, originated from five sub-districts including Aranyik, Hua Ro, Nai Muang, Ban Krang and Tha Pho, were used in the bioassays following the procedures of World Health Organization. For larval bioassay, the late third or early fourth-instar larvae were tested with different concentrations of temephos. For adult bioassay, the females were exposed to 0.75 % permethrin or 0.05% deltamethrin. LC50 value and mortality rate were analyzed to compare the insecticide susceptibility of the larvae and the adults in each area, respectively. RESULTS The LC50 value of temephos for the larvae from Aranyik, Hua Ro, Nai Muang, Ban Krang and Tha Pho sub-districts was 0.017, 0.017, 0.026, 0.061, and 0.113 ppm, respectively. For permethrin, the highest mortality rate (86.84%) was found in the mosquitoes from Aranyik but the others were more resistant with the lower mortality rates (16.00-42.67%). The adult mortality rates after exposing to deltamethrin were higher (82.34-98.67%) in all areas. CONCLUSIONS Ae. aegypti larvae were still susceptible to temephos. Conversely, most tested adults tended to resist the permethrin and deltamethrin.

Variation in the Time of Colonization of Broiler Carcasses by Carrion Flies in Nakhonsawan Province, Thailand

Abstract Carrion flies are the primary insects colonizing vertebrate carrion; however, limited information is available on the variation in the time of colonization (TOC) as related to time of placement (TOP) and time of death (TOD), particularly in Thailand. Three seasonal sets of nine broiler carcasses (euthanized and placed in field within 0.5 h after death) were placed in mesh enclosures within a disturbed deciduous dipterocarp forest at Nakhonsawan Province, upper-central Thailand, for 3 d to determine the colonization time by carrion flies. In total, 21,536 arthropods were collected using traps placed over each carcass. Carrion flies of the family Calliphoridae, Muscidae, and Sarcophagidae predominated (93.42%). Of these, Chrysomya megacephala (F.) (Diptera: Calliphoridae) and Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae) were the dominant species being 36.18% and 35.36%, respectively, across season. These species arrived immediately (5 min) after placement of the carrion in the field during the rainy season, while they were delayed 1–2 h during the dry season. Chrysomya megacephala, C. rufifacies, and Parasarcophaga dux (Thomson) colonized the remains. Time of colonization by C. megacephala and C. rufifacies occurred mostly at ~1600–1700 hours (10–11 h after placement) for all seasons. In contrast, TOC by P. dux was delayed for 1 d during rainy and dry season. These results mark the first record of carrion fly colonization in this area and also may deserve important information for the further study as they demonstrate time of colonization differs from TOP and most importantly TOD.

Morphological Characteristics of Terminalia of the Wasp-Mimicking Fly, Stomorhina discolor (Fabricius)

Stomorhina discolor (Fabricius), a species of blow fly that mimics wasps, is distributed worldwide, but detailed information about characteristics of its adult terminalia is incomplete. To help fill this gap in the information, the morphology of adult stages of S. discolor was investigated using light microscopy and scanning electron microscopy. Observations using the light microscope revealed unique characteristics of the male genitalia that are markedly different from other blow fly species. More morphological detail, including observation of several sensilla (e.g., sensilla trichoid and sensilla basiconica) along the male terminalia and female ovipositor, was seen under the scanning electron microscope. These details can be taxonomically valuable for identifying males and females of S. discolor and may help address matters concerning copulation in this species.

Larvicidal efficacy of crude and fractionated extracts of Dracaena loureiri Gagnep against Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus mosquito vectors

Objective: To evaluate the larvicidal efficacy of crude and fractionated extracts of Dracaena loureiri endocarp against Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus mosquitos. Methods: Larvicidal activity was tested according to World Health Organization standard protocol. The third-stage larvae of each mosquito species were exposed to various concentrations of Dracaena loureiri crude extract and six groups of Dracaena loureiri fractionated extracts (RC-DT 009–014). Larval mortality rates were observed after 24 h and 48 h of exposure. Then, a computerized probit analysis of the mortality data was performed to determine lethal concentration 50 (LC50) and lethal concentration 90 values. Results: Anopheles minimus larvae (24-h LC50 77.88 mg/L) had the highest susceptibility to crude extract, whereas others (Aedes aegypti, 24-h LC50 224.73 mg/L; Aedes albopictus, 24-h LC50 261.75 mg/L; and Culex quinquefasciatus, 24-h LC50 282.86 mg/L) were significantly less susceptible. The most effective groups of fractionated extracts were RC-DT 012 and RC-DT 013. The mosquito species most susceptible to fractionated extracts was Culex quinquefasciatus, with 24-h LC50 values of 0.66 and 0.94 mg/L for RC-DT 012 and RC-DT 013, respectively. Conclusions: The larvicidal activity of fractionated extracts is more effective than that of crude extract against all tested mosquito species. For the most effective alternative larvicide, purification and a phytochemical constituent analysis must be performed.

Prevalence and factors of head lice infestation among primary school students in Northern Thailand

Abstract Objective To survey the prevalence of head lice infestation among primary school students in Chiang Rai Province, Northern Thailand from November 2015 to February 2016 and to investigate factors involving these infestations. Methods A total of 703 students were checked for head lice infestations. The data were analyzed by using descriptive statistics i.e. frequencies, percentages and correlations [odds ratio (OR) and 95% confidence interval (CI)]. Results The overall percentage of head lice infestations was 15.1%. No infestation was found in boy students. The prevalence of head lice infestation was significantly associated with itching on a scalp (OR = 5.206, 95% CI = 3.116–8.696), having dirty fingernails (OR = 2.019, 95% CI = 1.217–3.352), wearing dirty clothes (OR = 4.532, 95% CI = 1.651–12.438), having history of head lice infestations (OR = 3.998, 95% CI = 2.174–7.356) and family member having history of head lice infestations (OR = 1.997, 95% CI = 1.092–3.651). Conclusions Pediculosis capitis is still public health problem in this region. Effective treatment is required to control this infestation in urgent.