Carrion flies (Insecta: Diptera) found on human cadavers in Chiba prefecture, Honshu, Japan, with the first record of Fannia prisca from a human corpse.

Forensic entomology (FE) involves the collection and analysis of necrophagous insects and arthropods for postmortem interval estimation based on their growth and succession. In Japan, research on FE is limited, and the predominant species of necrophagous insects and their distribution have not been clarified. The present study aimed to clarify the actual situation of insects, targeting Diptera collected from human cadavers, in Chiba Prefecture. We targeted the cases for which specimens could be collected from forensic autopsies conducted at the Legal Medicine Department of Chiba University Graduate School of Medicine or from human bodies handled by the Chiba Police in 2019. We specifically chose cases from which adult insects could be bred. Fifty cases were applicable, and 47 cases were successful in rearing adult flies. Sixteen species of Diptera were identified: six species of Calliphoridae, four species of Sarcophagidae, and a few species of Fanniidae, Muscidae, Phoridae, Piophilidae, and Stratiomyidae. The most frequently observed species were Lucilia sericata (20 cases) and Chrysomya pinguis (18 cases). For the first time, globally, we recorded the presence of Fannia prisca in a human cadaver. In conclusion, this is the first report to reveal the presence of carrion flies in human death cases in Chiba Prefecture, Japan. Research in FE is important in the context of the Japanese natural and social environment since this can aid forensic investigations conducted by authorities.

Daily and seasonal variation of muscid flies (Diptera: Muscidae) in Chiang Mai province, northern Thailand.

Flies of the family Muscidae, or muscids, are of medical and veterinary importance worldwide due to their recognition as nuisance pests and myiasis-producing agents. Effective control of muscids requires biological information on population dynamics daily and across seasons. In this study, such patterns were investigated in three different microhabitats (e.g., forest area, palm plantation and longan orchard) in a suburban area of Chiang Mai Province, northern Thailand. Adult fly samplings were conducted for 24-h intervals using semiautomatic traps and 1-day old beef offal as bait. Samplings were carried out twice per month from July 2013 to June 2014. A total of 3,419 muscids were trapped, comprising nine species, with Musca domestica Linnaeus accounting for the majority (n = 1,329; 38.9%) followed by Hydrotaea spinigera Stein (n = 770; 22.5%) and Musca ventrosa Wiedemann (n = 740; 21.7%). The greatest overall abundance was in the longan orchard location (n = 1,508; 44.1%). Community structure peaked during the rainy season (mid-May to mid-Oct). Peak activity during the day was late morning (9.00 to 12.00 h) for M. domestica, early morning (6.00 to 9.00 h) for H. spinigera, and early afternoon (12.00 to 15.00 h) for M. ventrosa. Temperature had no significant effect on the abundance of M. domestica (rs= -0.030, p = 0.576) or H. spinigera (rs = 0.068, p = 0.200), but had a weak negative correlation with M. ventrosa (rs = -0.238, p = 0.0001). Relative humidity had a weak negative correlation with M. domestica (rs = -0.263, p = 0.0001), H. spinigera (rs = -0.107, p = 0.043) and M. ventrosa (rs = -0.344, p = 0.0001). More females (n = 2,078) were trapped than males (n = 761). These results provide baseline information of daily and seasonal dynamic activity of muscid flies under natural conditions, which is the prerequisite information for effective control measures.

Mitochondrial DNA-Based Identification of Forensically Important Flesh Flies (Diptera: Sarcophagidae) in Thailand.

Flesh flies (Sarcophagidae) are necrophagous insects initially colonizing on a corpse. The species-specific developmental data of the flies collected from a death scene can be used to estimate the minimum postmortem interval (PMImin). Thus, the first crucial step is to correctly identify the fly species. Because of the high similarity among species of flesh flies, DNA-based identification is considered more favorable than morphology-based identification. In this study, we demonstrated the effectiveness of combined sequences (2216 to 2218 bp) of cytochrome c oxidase subunit I and II genes (COI and COII) for identification of the following 14 forensically important flesh fly species in Thailand: Boettcherisca nathani Lopes, Fengia ostindicae (Senior-White), Harpagophalla kempi (Senior-White), Liopygia ruficornis (Fabricius), Lioproctia pattoni (Senior-White), Lioproctia saprianovae (Pape & Bänziger), Parasarcophaga albiceps (Meigen), Parasarcophaga brevicornis (Ho), Parasarcophaga dux (Thomson), Parasarcophaga misera (Walker), Sarcorohdendorfia antilope (Böttcher), Sarcorohdendorfia inextricata (Walker), Sarcorohdendorfia seniorwhitei (Ho) and Seniorwhitea princeps (Wiedemann). Nucleotide variations of Thai flesh flies were evenly distributed throughout the COI-COII genes. Mean intra- and interspecific variations ranged from 0.00 to 0.96% and 5.22% to 12.31%, respectively. Using Best Match (BM) and Best Close Match (BCM) criteria, identification success for the combined genes was 100%, while the All Species Barcodes (ASB) criterion showed 76.74% success. Maximum Likelihood (ML) and Bayesian Inference (BI) phylogenetic analyses yielded similar tree topologies of monophyletic clades between species with very strong support values. The achieved sequences covering 14 forensically important flesh fly species including newly submitted sequences for B. nathani, F. ostindicae and S. seniorwhitei, can serve as a reliable reference database for further forensic entomological research in Thailand and in other areas where those species occur.

Spatial Distribution of Forensically Significant Blow Flies in Subfamily Luciliinae (Diptera: Calliphoridae), Chiang Mai Province, Northern Thailand: Observations and Modeling Using GIS.

Blow flies of the subfamily Luciliinae (Diptera: Calliphoridae) are one of the main forensically important subfamilies globally. In addition to being used to estimate the minimum post-mortem interval (PMImin), assuming colonization occurred after death, blow fly specimens found infesting a human corpse are used to determine if the corpse was relocated or if the individual ingested narcotics prior to death. The presence of these blow flies in a given area is strongly influenced by abiotic and biotic factors, such as temperature, elevation, and habitat. Having this information, along with geographical distributions and the characteristics of preferred habitats, is necessary to better understand the biology of this group. This study aimed to characterize the spatial distribution of Luciliinae throughout 18 sampling sites within six ecozones (disturbed mixed deciduous forest, mixed deciduous forest, mixed orchard, paddy field, lowland village, and city/town) in central Chiang Mai Province, northern Thailand over one year (May 2009⁻May 2010). The purpose of the study was to elucidate the relationship of blow fly species composition with environmental abiotic factors (e.g., temperature, relative humidity, light intensity), and to predict the distribution of the common species within this subfamily using GIS. Adult collections were performed biweekly, baited with one-day-old beef offal. A total of 2331 Luciliinae flies trapped, comprising eight species, of which the four predominant species were Hemipyrellia ligurriens (Wiedemann) (n = 1428; 61.3%), Lucilia porphyrina (Walker) (n = 381; 16.3%), Hemipyrellia pulchra (Wiedemann) (n = 293; 12.6%), and Lucilia papuensis Macquart (n = 129; 5.5%). Population density across species varied seasonally, peaking in August 2009 coinciding with the rainy season. Predicting population composition was based on a model developed with ArcGIS 9.2, which utilized environmental variables (temperature, relative humidity, and light intensity) in conjunction with abundance data. Models indicated H. ligurriens had the most widespread geographic distribution, while H. pulchra was predicted to occur largely in mixed orchards and lowland villages. Lucilia porphyrina and L. papuensis were less widespread, restricted mainly to mixed deciduous forest. This model, along with knowledge of forensic information, may be useful under certain investigations where the corpse may have been relocated.

Predicting Geographic Distribution of Forensically Significant Blow Flies of Subfamily Chrysomyinae (Diptera: Calliphoridae) in Northern Thailand.

Blow flies (Diptera: Calliphoridae) are carrion-breeding flies that are commonly used as evidence in forensic investigation. An adequate knowledge of ecological and geographical data of blow fly has a direct application in forensic science, as far as estimating time of colonization or corpse relocation. The aim of this study was to evaluate the occurrence of four species of Chrysomyinae (Chrysomya pinguis, Chrysomya chani, Chrysomya villeneuvi, and Ceylonomyia nigripes) across six land use types in central Chiang Mai, northern Thailand. Eighteen study sites were selected for sampling across three districts of Chiang Mai province (Mueang Chiang Mai, Mae Rim, and Hang Dong). Adult flies were collected every two weeks using a funnel trap baited with 1-day tainted beef offal. The predicted geographic distributions of forensically important blow fly species were modeled using the computer program ArcGIS, based on selected climatic variables (temperature, relative humidity, and light intensity) recorded at study sites. During the study period, 1298 adult flies were collected, with peak fly occurrence during summer (April⁻May). Seasonal fluctuation patterns varied depending on fly species. Climatic factors displayed diverse impact on associated fly populations. Identified species were restricted mainly to mixed deciduous forests (MDF) especially in the mountainous area. None of these flies were trapped in an urban area.

Morphology of immature stages of blow fly, Lucilia sinensis Aubertin (Diptera: Calliphoridae), a potential species of forensic importance.

Lucilia sinensis Aubertin (Diptera: Calliphoridae) is a blow fly species of potential forensic importance since adults are attracted to, and colonize, decomposing vertebrate remains. Blow fly larvae associated with human corpses can be useful evidence in forensic investigations; however, their use is dependent in most cases on proper species identification and availability of developmental data. For identification, morphological information on each life stage is traditionally used. We used scanning electron microscopy (SEM) to examine the ultrastructure of eggs, all instars, and puparia, of L. sinensis. The important characteristics used to differentiate L. sinensis from other species are provided. Distinctive features of the eggs are the slight widening median area extending almost the entire length. The last abdominal segment of the first instar bears elongated outer ventral tubercles along the rim of the last abdominal segment. These tubercles, as well as the well developed median and outer dorsal tubercles, are more prominent in the second and third instars. The surface integument of the tubercles is equipped with circular rows of microtrichia. Pairs of inner dorsal tubercle are absent. Each anterior spiracle is comprised of 9-12 papillae arrange in a single row in the second and third instars. As for the third instar, the dorsal spines between the first and second thoracic segments are delicate, narrow, small, and close together (as row or set). The peristigmatic tufts adjacent to the posterior spiracle of the third instar are moderately branches of short, fine hairs, but minute in puparia. In conclusion, the prominent outer ventral tubercle in all instars and puparia is a new diagnostic feature of L. sinensis and helpful in differentiating it from other Lucilia species that are forensically important. The description of immature L. sinensis in this study will be useful for forensic entomologists in countries where this species exists.

Field evaluation of a semi-automatic funnel trap targeted the medically important non-biting flies.

Bait-trapping is a useful approach for monitoring fly population dynamics, and it is an effective tool for physical control of pest species. The aim of this study was to test a newly developed semi-automatic funnel fly trap with some modifications of the former prototype fly trap to study medically important fly population density. The efficacy of the semi-automatic funnel trap was assessed by field sampling during July 2013-June 2014 using 1-day tainted beef offal as bait. The modified semi-automatic funnel traps were able to capture a total of 151,141 adult flies, belonging to the families: Calliphoridae (n=147,248; 97.4%), Muscidae (n=3,124; 2.1%) and Sarcophagidae (n=769; 0.5%), which are the medically important fly species. Among the total of 35 species collected, Chrysomya megacephala (Diptera: Calliphoridae) (n=88,273; 59.95%), Musca domestica (Diptera: Muscidae) (n=1,324; 42.38%) and Boettcherisca peregrina (Diptera: Sarcophagidae) (n=68; 33.01%) were the predominant species of each family. High number of flies was captured in forest area, representing 42.47% (n=64,197) of total specimens. Female flies were trapped more than male with total sex ratio of 0.37 male/female. Flies were trapped throughout the year with peak population in summer. Peak activity was recorded in the afternoon (12.00-18.00h). In summary, the modified semi-automatic funnel fly trap can be used for field collection of the adult fly. By setting the timer, population dynamics, diversity, and periodic activity of adult flies were determined.

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.

The Importance of Habitat in the Ecology of Decomposition on Rabbit Carcasses in Malaysia: Implications in Forensic Entomology.

The stages of decomposition and the faunal succession on rabbit carcasses in three different habitats, namely jungle, rural, and highland areas, were studied. Three New Zealand White rabbit (Oryctolagus cuniculus) carcasses weighing ∼2 kg were sampled daily until the decomposition process was completed. Representative specimens of adult flies, larvae, pupa, and mites were collected from the carcasses and processed in the laboratory. There were differences in decomposition rate and faunal succession between the carcasses. The fastest rate of decomposition was recorded in rural area, and the slowest rate of decomposition was recorded in highland area. The carcasses exhibited the same pattern of colonization by adult flies, but the dominant species of larvae and adult flies on each carcass in specific habitats were different. The primary species of flies recorded in jungle were Chrysomya megacephala F., Chrysomya rufifacies (Macquart), Chrysomya chani Kurahashi, Chrysomya villenuevi Patton, Chrysomya nigripes Aubertin, Chrysomya pinguis (Walker), Hemipyrellia ligurriens (Wiedemann), Hemipyrellia tagaliana (Bigot), Hypopyiopsis fumipennis (Walker), Hypopygiopsis violacea (Macquart), and Hydrotaea spinigera Stein represented by both adults and larvae. Musca domestica L., Atherigona sp., Lioproctia pattoni (Senior-White), Lioproctia saprianovae Pape & Bänziger, and Seniorwhitea princeps (Wiedemann) were represented by adults only. The biodiversity of flies in the rural area were C. megacephala, C. rufifacies, H. ligurriens, Fannia canicularis L., Hydrotaea chalcogaster (Wiedemann), and Hyd. spinigera represented by both adults and larvae, meanwhile M. domestica, Atherigona sp., Boettcherisca peregrina (Robineau-Desvoidy), Parasarcophaga taenionota Wiedemann, Parasarcophaga scopariiformis Senior-White, and S. princeps were represented by adults only. The species of flies collected in the highland area were Lucilia porphyrina (Walker), C. megacephala, C. rufifacies, C. villenuevi, C. pinguis, H. ligurriens, Hyd. spinigera, Hyd. chalcogaster, F. canicularis, and Boettcherisca highlandica Kurahashi & Tan represented by both adults and larvae, whereas C. nigripes, Chrysomya thanomthini Kurahashi & Tumrasvin, M. domestica, Atherigona sp., Parasarcophaga albiceps Meigen, P. taenionota, Sepsidae, Phoridae, and Millichidae were represented by adults only. Faunal succession followed the sequence of dominant flies, i.e., Calliphoridae, Sarcophagidae, Muscidae, Sepsidae, and lastly Stratiomyidae for jungle, or Sepsidae for rural and highland studies. Mites, from suborders Mesostigmata, Prostigmata, Astigmatina, and Oribatida, were also recovered throughout decomposition, which could be used for future implementation in forensic investigations. The data obtained from this study could provide more accurate indicators for local forensic scientists in solving criminal cases especially on the determination of time and primary location of death.

Keys to the blow flies of Taiwan, with a checklist of recorded species and the description of a new species of Paradichosia Senior-White (Diptera, Calliphoridae).

BLOW FLIES (DIPTERA: Calliphoridae) show a great diversity in behavior and ecology, play important roles in ecosystems, and have medical and forensic importance to humans. Despite this, the taxonomy and classification of Taiwan's Calliphoridae have rarely been studied. In this study, specimens of Taiwanese calliphorids were collected and carefully studied, and all 76 species recorded in Taiwan are listed following the identification keys. Dichotomous keys to all subfamilies, tribes, genera, and species of blow flies recorded in Taiwan are provided, including 16 species that are newly recorded from Taiwan. In addition, one new species of the genus Paradichosia Senior-White is described and illustrated. We also discuss the morphological differences between the specimens of Silbomyia hoeneana Enderlein collected from China and Taiwan, a species that has only been found previously in Southern China.

Species composition of carrion blow flies in northern Thailand: altitude appraisal.

Distribution and occurrence of blow flies of forensic importance was performed during 2007 and 2008 in Chiang Mai and Lampang Provinces, northern Thailand. Surveys were conducted in forested areas for 30 minutes using a sweep net to collected flies attracted to a bait. A total of 2,115 blow flies belonging to six genera and 14 species were collected; Chrysomya megacephala (Fabricius) (44.7%), C. pinguis (Walker) (15.1%), C. chani Kurahashi (9.3%), C. thanomthini Kurahashi & Tumrasvin (0.3%); Achoetandrus rufifacies (Macquart) (10.5%), A. villeneuvi (Patton) (2.2%); Lucilia papuensis Macquart (2.2%), L. porphyrina (Walker) (12.4%), L. sinensis Aubertin (0.7%); Hemipyrellia ligurriens (Wiedemann) (1.3%), H. pulchra (Wiedemann) (0.1%); Hypopygiopsis infumata (Bigot) (0.6%), Hy. tumrasvini Kurahashi (0.2%) and Ceylonomyia nigripes Aubertin (0.4%). Among them, C. megacephala was the predominant species collected, particularly in the summer. The species likely to prevail in highland areas are C. pinguis, C. thanomthini, Hy. tumrasvini, L. papuensis and L. porphyrina.

Two new records of Isomyia paurogonita Fang and Fan, 1986 and Sumatria latifrons Malloch, 1926 (Diptera: Calliphoridae) from northern Thailand, with revised key to the species of Isomyia.

During the annual fly survey at Doi Nang Kaew in Doi Saket District, Chiang Mai Province of Thailand in 2011, Isomyia paurogonita Fang & Fan, 1986 (Diptera: Calliphoridae) and Sumatria latifrons Malloch, 1926 (Diptera: Calliphoridae) were collected for the first time in Thailand. They are the rare species of the subfamily Rhiniinae (tribe Cosminini). Prior to this finding, fifteen species of Isomyia and two species of Sumatria were recorded from Thailand. Therefore, 96 blow fly species have been found in this country. These new locality records of both flies are very important for further research on their biology and ecology in Thailand.

Impact of abiotic factor changes in blowfly, Achoetandrus rufifacies (Diptera: Calliphoridae), in northern Thailand.

Understanding how medically important flies respond to abiotic factor changes is necessary for predicting their population dynamics. In this study, we investigated the geographical distribution of the medically important blowfly, Achoetandrus rufifacies (Macquart) (Diptera: Calliphoridae), and ascertained the response to climatic and physio-environmental factors in Chiang Mai, northern Thailand. Adult fly surveys were carried out every 2 weeks from May 2009 to May 2010 at 18 systematically randomized study sites in three districts of Chiang Mai province (Mueang Chiang Mai, Mae Rim, and Hang Dong), using reconstructable funnel traps with 1-day tainted beef offal as bait. During the study period, 8,861 adult A. rufifacies were captured, with peak densities being observed at the end of winter (i.e., late February) and throughout most of the summer (May to March). Population density had a weak but significant (α = 0.05) positive correlation with temperature (r = 0.329) and light intensity (r = 0.231), and a weak but significant (α = 0.05) negative correlation with relative humidity (r = -0.236). From the six ecological land use types (disturbed mixed deciduous forest, mixed deciduous forest, mixed orchard, lowland village, city town, and paddy field), greater fly densities were observed generally in the disturbed mixed deciduous forest and lowland village, but not in the paddy fields. In conclusion, A. rufifacies are abundant from the end of winter and throughout most of the summer in northern Thailand, with population density being weakly positively correlated with temperature and light intensity, but weakly negatively correlated with relative humidity. The greatest densities of this fly species were collected in disturbed mixed deciduous forest and lowland village land uses. The prediction of annual and season specific distributions of A. rufifacies were provided in each season and all-year patterns using a co-kriging approach (ArcGIS9.2).

Evaluation of an I-box wind tunnel model for assessment of behavioral responses of blow flies.

The behavioral response of flies to olfactory cues remains the focus of many investigations, and wind tunnels have sometimes been employed for assessment of this variable in the laboratory. In this study, our aim was to design, construct, and operate a new model of I-box wind tunnel with improved efficacy, highlighting the use of a new wind tunnel model to investigate the behavioral response of the medically important blow fly, Chrysomya megacephala (Fabricius). The I-box dual-choice wind tunnel designed for this study consists of seven conjoined compartments that resulted in a linear apparatus with clear glass tunnel of 30 × 30 × 190 cm ended both sides with wooden "fan compartments" which are equipped with adjustable fans as wind source. The clear glass tunnel consisted of two "stimulus compartments" with either presence or absence (control) of bait; two "trap compartments" where flies were attracted and allowed to reside; and one central "release compartment" where flies were introduced. Wind tunnel experiments were carried out in a temperature-controlled room, with a room light as a light source and a room-ventilated fan as odor-remover from tunnel out. Evaluation of testing parameters revealed that the highest attractive index was achieved with the use of 300 g of 1-day tainted pork scrap (pork meat mixed with offal) as bait in wind tunnel settings wind speed of 0.58 m/s, during 1.00-5.00 PM with light intensity of 341.33 lux from vertical light and 135.93 lux from horizontal light for testing a group of 60 flies. In addition, no significant response of well-fed and 24 h staved flies to this bait under these conditions was found. Results of this study supported this new wind tunnel model as a suitable apparatus for investigation of behavioral response of blow flies to bait chemical cues in the laboratory.

Occurrence of blow fly species (Diptera: calliphoridae) in Phitsanulok Province, Northern Thailand.

Based on the current forensic importance of blow flies (Diptera: Calliphoridae), their biological aspects have been studied increasingly worldwide. The blow fly fauna in Phitsanulok Province, Northern Thailand was studied from May 2009 to April 2010 in the residential, agricultural, mountainous and forested areas of Muang, Wat Bot, Nakhon Thai and Wang Thong districts, respectively, in order to know the occurrence of blow flies in this province. Collections were carried out monthly using commercial funnel fly traps and sweeping methods, with 1-day tainted pork viscera as bait. Identification of adult blow flies exhibited 14 634 specimens, comprising of 5 subfamilies, 14 genera and 36 species. Chrysomya megacephala (Fabricius, 1794) and Achoetandrus rufifacies (Macquart, 1843) were the most and second most abundant species trapped, respectively. These two species of carrion flies prevailed in all the types of land investigated. We calculated and compared the diversity indices, species evenness and richness, and similarity coefficients of the blow fly species in various areas. The data from this study may be used to identify the potential of forensicallyimportant fly species within Phitsanulok Province and fulfill the information on blow fly fauna in Thailand.

Reproductive Organ of Blow Fly, Chrysomya megacephala (Diptera: Calliphoridae): Ultrastructural of Testis.

This work presents the ultrastructure of testis of the medically important blow fly, Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) using light microscopy and electron microscopy. Reproductive organ of males was dissected to determine the testis in the pupal stage, 3-day-old flies and 7-day-old flies and observed under scanning electron microscopy (SEM) and transmission electron microscopy (TEM). SEM displayed a smooth surface which is occasionally penetrated by tracheoles. TEM of the testis in the pupal stage presents the thick testis wall covering underdeveloped cells containing a variable size of an electron-dense globule. For the 3-day-old males, the testicular wall is formed by an external layer, a peritoneal sheath, a muscular layer, a basement membrane, and a follicular epithelium. Follicular epithelium presented developing spermatozoa. Regarding the 7-day-old males, development of spermatozoa is apparent, displaying nucleus, centriolar adjunct, axoneme, and mitochondrial derivatives, with the 9 + 9 + 2 microtubule pattern of axoneme.

Effects of human contraceptive on reproduction and offspring in Chrysomya megacephala.

OBJECTIVE: To investigate the effect of human contraceptive (HC) as ability to suppress the reproductive success of blow fly, Chrysomya megacephala (Fabricius) (C. megacephala) and offspring under controlled laboratory conditions. METHODS: Adult C. megacephala were fed with low (0.036 mg/mL) and high dose (0.072 mg/mL) HC (Microgest®, Thailand), containing levonorgestrel and ethinyl estradiol, in their drinking water for 7 days. Three experiments were set; experiment I with fed only in parental males, experiment II with fed only in parental females and experiment III with fed in both males and females. All experiments were then maintained for 3 generations after crossing and inbreeding. RESULTS: A lower ovariole production and less fully mature ovarioles were evident in F1, F2 and F3 than control when parent males, females and both had been fed with high dose HC. Cellular changes during spermatogenesis in F1, F2 and F3 testes was confirmed using transmission electron microscopy (TEM), showing the low level of condensed chromatin, necrotic chromatin, irregularities and degenerated nuclear envelope in the nucleus. In the cytoplasm, mitochondrial swelling, rough endoplasmic reticulum swelling as well as vacuolated cytoplasm were noticed. As for the sperm per se, we found the degenerated nuclei and/or incomplete mitochondrial derivative, axoneme and vacuolated flagella. Regarding deformity in F1, F2 and F3 ovariole, ultrastructural alteration observed by scanning electron microscopy (SEM) included malformations involving fragile enveloping peritoneal sheath, cracked ovarioles, peel away chorion, crumbled eggshell and incomplete development; whereas TEM presented malformed and disorganized mass of cells, proteic yolk granules and vacuolated vesicles. CONCLUSIONS: Administer of HC to adult C. megacephala caused ovariole reduction, less matured ovariole and affected cellular changes in testes and ovariole of offspring up to F3.

Blow Flies Were One of the Possible Candidates for Transmission of Highly Pathogenic H5N1 Avian Influenza Virus during the 2004 Outbreaks in Japan.

The 2003-2004 H5N1 highly pathogenic avian influenza (HPAI) outbreaks in Japan were the first such outbreaks in 79 years in Japan. Epidemic outbreaks have been occurring in Southeast Asia, with the most recent in 2010. Knowledge of the transmission route responsible for the HPAI outbreaks in these countries remains elusive. Our studies strongly suggested that field and laboratory studies focusing on mechanical transmission by blow flies should be considered to control H5N1 avian influenza outbreaks, in particular in epidemic areas, where there are high densities of different fly species throughout the year. In this paper, we review these field and laboratory entomological studies and discuss the possibility of blow flies transmitting H5N1 viruses.

Three new species of shoot fly, Atherigona spp., from northern Thailand.

Three new species of shoot fly, Atherigona Rondani (subgenus Acritochaeta Grimshaw) (Diptera: Muscidae), are described from northern Thailand, based on morphological characteristics of males. Unique features of A. komi sp. n. include a distinct spiral groove on the dorsal aspect of the fore femur and two dark apical wing spots, whereas A. chiangmaiensis sp. n. is recognized by the presence of one large patch on the apical wing spot, appearing as a large and smaller wave-shaped patch, and no distinct pattern on tergites. A. thailandica sp. n. displays a remarkable dark boomerang-shaped patch along the wing margin and fore femur, with two rows of long hairs on the dorsal surface. Male terminalia are also different in the new species, showing distinctive characteristics. This paper also presents five newly recorded species in Thailand; Atherigona maculigera Stein, Atherigona ovatipennis vietnamensis Shinonaga et Thinh, Atherigona pallidipalpis Malloch, Atherigona seticauda Malloch, and Atherigona setitarsus Shinonaga et Thinh. A key is provided for the adult males of Atherigona recorded in Thailand, all belonging to the subgenus Acritochaeta, except for A. soccata Rondani.

DNA-based characterisation and classification of forensically important flesh flies (Diptera: Sarcophagidae) in Malaysia.

Insect larvae and adult insects found on human corpses provide important clues for the estimation of the postmortem interval (PMI). Among all necrophagous insects, flesh flies (Diptera: Sarcophagidae) are considered as carrion flies of forensic importance. DNA variations of 17 Malaysian, two Indonesian and one Japanese flesh fly species are analysed using the mitochondrial COI and COII. These two DNA regions were useful for identifying most species experimented. However, characterisation of the species was not sufficiently made in the case of Sarcophaga javanica. Seventeen Malaysian species of forensic importance were successfully clustered into distinct clades and grouped into the six species groups: peregrina, albiceps, dux, pattoni, princeps and ruficornis. These groups correspond with generic or subgeneric taxa of the subfamily Sarcophaginae: Boettcherisca, Parasarcophaga, Liosarcophaga, Sarcorohdendorfia-Lioproctia, Harpagophalla-Seniorwhitea and Liopygia. The genetic variations found in COI and COII can be applied not only to identify the species of forensic importance, but also to understand the taxonomic positions, generic or subgeneric status, of the sarcophagine species.