Multi-epitope protein production and its application in the diagnosis of opisthorchiasis.

BACKGROUND: Opisthorchiasis and cholangiocarcinoma (CCA) continue to be public health concerns in many Southeast Asian countries. Although the prevalence of opisthorchiasis is declining, reported cases tend to have a light-intensity infection. Therefore, early detection by using sensitive methods is necessary. Several sensitive methods have been developed to detect opisthorchiasis. The immunological detection of antigenic proteins has been proposed as a sensitive method for examining opisthorchiasis. METHODS: The Opisthorchis viverrini antigenic proteins, including cathepsin B (OvCB), asparaginyl endopeptidase (OvAEP), and cathepsin F (OvCF), were used to construct multi-antigenic proteins. The protein sequences of OvCB, OvAEP, and OvCF, with a high probability of B cell epitopes, were selected using BepiPred 1.0 and the IEDB Analysis Resource. These protein fragments were combined to form OvCB_OvAEP_OvCF recombinant DNA, which was then used to produce a recombinant protein in Escherichia coli strain BL21(DE3). The potency of the recombinant protein as a diagnostic target for opisthorchiasis was assessed using immunoblotting and compared with that of the gold standard method, the modified formalin-ether concentration technique. RESULTS: The recombinant OvCB_OvAEP_OvCF protein showed strong reactivity with total immunoglobulin G (IgG) antibodies against light-intensity O. viverrini infections in the endemic areas. Consequently, a high sensitivity (100%) for diagnosing opisthorchiasis was reported. However, cross-reactivity with sera from other helminth and protozoan infections (including taeniasis, strongyloidiasis, giardiasis, E. coli infection, enterobiasis, and mixed infection of Echinostome spp. and Taenia spp.) and no reactivity with sera from patients with non-parasitic infections led to a reduced specificity of 78.4%. In addition, the false negative rate (FNR), false positive rate (FPR), positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy were 0%, 21.6%, 81.4%, 100%, and 88.9%, respectively. CONCLUSIONS: The high sensitivity of the recombinant OvCB_OvAEP_OvCF protein in detecting opisthorchiasis demonstrates its potential as an opisthorchiasis screening target. Nonetheless, research on reducing cross-reactivity should be undertaken by detecting other antibodies in other sample types, such as saliva, urine, and feces.

Effect of cold argon plasma on eggs of the blow fly, Lucilia cuprina (Diptera: Calliphoridae).

Non-thermal plasma has been used in many medical applications, including treatment of living cells, blood coagulation, wound healing, and sterilization. The process uses an environmentally friendly gas (e.g., argon, helium, oxygen, nitrogen, or hydrogen) to destroy bacteria cells with no serious adverse effect on humans or animals. However, information on the effect of argon plasma on blow fly eggs is lacking. In this study, we explored the ability of cold argon plasma to destroy the eggs of the Australian sheep blow fly, Lucilia cuprina (Wiedemann, 1830); its larvae are a myiasis-producing agent in both human and animals. We tested the effect of cold argon plasma exposure for 1, 2, 3 and 5min on L. cuprina eggs. Since the temperature of cold Ar plasma is around 30°C, to clarify the effect of temperature on the fly eggs, hot air from an electric dryer was tested for comparison. Cold argon plasma exposure in eggs significantly reduced the survival rates of second instar larvae at all exposures tested; the effects were time dependent, with a stronger effect at longer exposure (32% survival rate after a 1-min treatment; 20%, 2min; 20%, 3min; and 6%, 5min), compared to the control (86%). No significant differences were observed in larval survival rates from eggs treated with hot air (80-84%, after 1- to 5-min treatments) versus the control (86%). These results were supported by observing the treated eggshells under a scanning electron microscope (SEM), we found noticeable aberrations only in the plasma treated groups. The emission spectrum of the argon gas discharge revealed emission lines of hydroxyl radicals at 309.1nm; these may cause the deterioration of the treated L. cuprina eggs. Our results have shown the possibility of using cold argon plasma in medical applications, in particular treating myiasis wounds.

Sterilization of blow fly eggs, Chrysomya megacephala and Lucilia cuprina, (Diptera: Calliphoridae) for maggot debridement therapy application.

Maggot debridement therapy (MDT) is an application of sterile laboratory-reared blow fly larvae to remove necrotic tissue and disinfect wounds for medical conditions. For effective application, the blow fly larvae used in the wound treatment are required to be in aseptic condition. Here, we report the results of a detailed assessment of bacteria and fungi isolated from the eggs of two blow fly species, Chrysomya megacephala (F.) and Lucilia cuprina (Wiedemann) before and after sterilization by disinfectants Chlorhex-C, povidone-iodine, and sodium hypochlorite. We also assess the survival ability of larvae and their sterility after the cleansing process. The results indicate that the isolated microorganisms from the control group of both the species consisted of 10 species of gram-positive bacteria, 21 species of gram-negative bacteria, and 4 species of yeast. As for sterility testing, the eggs and the larvae of C. megacephala were found to have been completely sterilized after being subjected to thioglycollate medium for 5 days, leading to aseptic larvae. By contrast, some microorganisms from the bacterial culture were still detected in the L. cuprina larvae treated with Chlorhex-C and povidone-iodine. The survival ability of the larvae in both the species was not significantly different between the treated and the control groups. Due to its high disinfection efficacy in destroying microorganisms in both the blow fly eggs, sodium hypochlorite is recommended for preparing sterile larvae before using MDT.

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.

Antibacterial Activity of Excretions-Secretions from Chrysomya megacephala Against Escherichia coli.

BACKGROUND: The blowfly, Chrysomya megacephala, is distributed worldwide. Previous studies found maggot excretions-secretions from other blowfly species inhibited pro-inflammatory response and antimicrobial activity. OBJECTIVE: This study aimed to test the bactericidal activity of excretions-secretions from C. megacephala larvae. MATERIAL AND METHOD: A total of 1,500 3-day-old larvae were used to collect excretions-secretions (ES) modified by the Barnes method. The bactericidal activity ofthe excretions-secretions was test by Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli using suitable liquid culture assay. Scanning electron microscope (SEM) was used to investigate the morphological change ofthe bacteria. RESULTS: E. coli were significantly inhibited by excretions-secretions from C. megacephala larvae. P. aeruginosa and S. aureus were not found to inhibit growth. CONCLUSION: The excretions-secretions from C. megacephala larvae may have a medical property for the inhibition of bacterial growth.

Isolation, Identification, and Evaluation of Novel Probiotic Strains Isolated from Feces of Breast-Fed Infants.

OBJECTIVE: To isolate, identify, and evaluate the probiotic properties of lactic acid bacteria (LAB) isolated from the feces of breast-fed infants. MATERIAL AND METHOD: The probiotic tests included investigation of hemolysis activity, survival in simulated gastrointestinal tract conditions (acid and bile salt tolerance), susceptibility to antibiotics, and ability to inhibit selected bacterial pathogens (Escherichia coli O157:H7, Vibrio cholerae and Salmonella enterica subsp enterica serovar Typhimurium). The bacterial species identification was performed by both carbohydrate utilization and partial 16S ribosomal RNA sequencing. RESULTS: Five of fifty LAB isolates (UBU-03, UBU-06, UBU-09, UBU-34, and UBU-37) showed good probiotic properties. These five isolates showed non-hemolysis type (gamma-hemolysis), susceptibility to all antibiotics tested except for vancomycin, ability to survive in the simulated gastrointestinal conditions of both acid and bile salt solution, and ability to inhibit growth of E. coli O157: H7 and V. cholerae. Bacterial species identification revealed that all five isolates were firmly identified as Lactobacillus rhamnosus species. CONCLUSION: The L. rhamnosus strains that were isolated and characterized in this study could be considered as probiotic strains, and then used for further probiotic characterization in human cell cultures or animal models.

Antibiotic resistance patterns of Enterococcus spp. isolated from Musca domestica and Chrysomya megacephala in ubon Ratchathani.

BACKGROUND: The housefly Musca domestica and the blowfly Chrysomya megacephala are found worldwide and are medically significant as mechanical vectors of various pathogens from unsanitary locations to food, resulting in diseases in humans. OBJECTIVES: This study aimed to test the antimicrobial activity against Enterococcus spp. isolated from M. domestica and C. megacephala by standard disk diffusion and minimal inhibitory concentration (MIC), and to study the potential of M. domestica and C. megacephala to transfer multi-drug resistant enterococcus to humans. MATERIAL AND METHOD: Seven hundred adult flies were collected from fresh-food markets, garbage piles, restaurants, school cafeterias, and rice paddy fields in Muang Ubon Ratchathani and Warinchamrap in Ubon Ratchathani Province. Antimicrobial susceptibility for Enterococcus spp. isolated from adult flies was performed by disk diffusion test and minimum inhibitory concentration (MIC) determination. RESULTS: One hundred and twenty isolates of Enterococcus spp. were taken from 67 M. domestica and 53 C. megacephala. Standard disk diffusion showed the Enterococcus spp. isolates exhibited susceptibility to ampilcillin (99.2%), chloramphenicol (74.20%), tetracycline (75.0%), vancomycin (50.8%), and erythromycin (42.5%). The MICs of antimicrobial agents for all isolates were < or = 0.25-8 microg/mL for vancomycin, 1- > 16 microg/mL for tetracycline, 4- > 16 microg/mL for chloramphenicol, and 0.5-8 microg/mL for ciprofloxacin. CONCLUSION: The study demonstrated the potential of M. domestica and C. megacephala to carry Enterococcus spp. Nine antimicrobial susceptibility patterns were obtained among the 120 enterococci isolates.

Isolation of Toxocara eggs from flies in Northeast Thailand.

BACKGROUND: Flies are natural carriers of pathogens and play a considerable role in the mechanical transmission of many pathogens, such as viruses, fungi, bacteria, and parasites, in various regions of the world. OBJECTIVE: To investigate the existence of Toxocara spp. eggs on two types of flies, the Oriental latrine fly, Chrysomya megacephala, and the housefly, Musca domestica, in Ubon Ratchathani, Northeast Thailand. MATERIAL AND METHOD: Flies were collected by the use of sterilized insect sweeping nets at five types of sites, fresh-food markets, garbage piles, restaurants, school cafeterias, and paddy fields from September 2010 to October 2011. After centrifugation, sediments from each fly species were examined for parasites under a light microscope. RESULTS: From the analysis of 555 C. megacephala and 439 M. domestica adult samples, Toxocara eggs were isolated from only the former species in all sites except for the restaurants in the Warinchamrap. No other helminthes and protozoa were found on the body surfaces of both fly species. CONCLUSION: The data suggest that C. megacephala is more likely to be a potential carrier of Toxocara eggs than M. domestica in areas frequented by humans in this region of Thailand.

Survey of the synanthropic flies associated with human habitations in ubon ratchathani province of northeast Thailand.

Synanthropic fly surveys were performed to determine the species composition and abundance in Ubon Ratchathani province in Northeast Thailand. Adult fly collections were conducted in various human habitations from two districts-Muang Ubon Ratchathani and Warinchamrap, at fresh-food markets, garbage piles, restaurants, school cafeterias, and rice paddy fields. Customized reconstructable funnel fly traps baited with 250 g of 1-day tainted beef were used for fly collections from September 2010-February 2011. A total of 3,262 flies were captured, primarily consisting of three families including: Calliphoridae (6 species), Muscidae (3 species), and Sarcophagidae (11 species). The blow fly, Chrysomya megacephala, and the house fly, Musca domestica, were the dominant species collected from both districts at all collection sites. C. megacephala was predominant in paddy fields, restaurants and garbage piles, while M. domestica was numerically dominant in fresh-food markets and school cafeterias. The current survey identified various species of synanthropic flies with close associations to humans and with the ability to transmit human pathogens in Ubon Ratchathani province; providing crucial information that may be used for developing control and sanitation management plans in this particular area.

Alimentary Canal of the Adult Blow Fly, Chrysomya megacephala (F.) (Diptera: Calliphoridae)-Part I: Ultrastructure of Salivary Glands.

The salivary gland ultrastructure of the adult male blow fly, Chrysomya megacephala (F.) (Diptera: Calliphoridae), was investigated at the ultrastructural level using light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The salivary glands are paired structures composed of a single median deferent duct bifurcated into two long, narrow efferent ducts connected to the coiled tubular glands. The SEM image of the gland surface revealed that the basal lamina is relatively smooth in general, but the whole surface appeared as a trace of rough swollen insertion by intense tracheal ramification. Ultrastructurally, the salivary gland is enclosed within the basal lamina, and interdigitation cytoplasmic extensions were apparent between the adjacent gland cells. The basement membrane appeared infoldings that is similar to the complex of the labyrinth channel. The cytoplasm characteristic of the gland revealed high activity, based on the abundance of noticeable secretory granules, either singly or in an aggregated reservoir. In addition, mitochondria were found to intersperse among rich parallel of arrays rough endoplasmic reticulum. Thick cuticle, which was well-delineated and electron dense, apically lined the gland compartments, with discontinuity of the double-layer cuticle revealing a trace of secretion discharged into the lumen. Gross anatomy of the adult salivary gland was markedly different from that of the third instar of the same species, and structural dissimilarity is discussed briefly.

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.

Forensically important flesh fly species in Thailand: morphology and developmental rate.

Forensically important flesh fly species in Thailand have been investigated for their larval morphology and developmental rate to increase the forensic entomology database in Thailand and nearby countries. Sarcophaga (Liosarcophaga) dux Thomson, Sarcophaga (Liopygia) ruficornis (Fabricius), and Sarcophaga (Boettcherisca) peregrina (Robineau-Desvoidy) are the species of greatest forensic importance. The third instars of these species are similar in morphological appearance, making it difficult or impossible to differentiate between them based on external morphological characteristics. This study compares these important characteristics and reveals that the most distinctive is the posterior spiracle, followed by the number of papillae on the anterior spiracle. For confirmation of these species, the unique characteristics of adult males are also shown for accurate identification. Both the third instar and adult male characteristics are keys to identifying these species. In addition, the developmental rate of S. dux was examined in northern Thailand during 2002-2003 under natural ambient temperature (approximately 24-28 degrees C) and a natural light/dark photoperiod (approximately 12:12 h), indicating relatively rapid larval development in summer (March-June), pre-pupae (stop moving) initiated at 72 h. In the rainy season (July-October), pre-pupae initiated at 72 or 96 h, whereas pre-pupae initiated at 96 h in winter.

Two new species of Sarcophaga s. lat. from Thailand with a key to species (Diptera: Sarcophagidae).

During a distribution survey of flies of medical importance around northern Thailand, we discovered two new species of flesh fly Sarcophaga (s. lat.) (Diptera: Sarcophagidae) that also are new to science. The species are Sarcophaga nanensis sp. nov. and Sarcophaga kiyokoae sp. nov. Adults of the former species was recovered from Pua River of Nan province (19 degrees 12'29" N, 100 degrees 56' 36" E) 442 m in northern Thailand, whereas the latter species was collected around Kuang River (19 degrees 0'3" N, 99 degrees 17'16" E) 551 m of the lowland forests of Chiang Mai province, northern Thailand. Although these two sarcophagid flies showed similarity in their male terminalia in having large aedeagi and rounded juxta, illustrations of the sternite 5, cercus, surstylus, and anterolateral margin of the aedeagus allow differentiation between them. A key is provided to the adult males of the flesh flies of medical importance in Thailand.

Ultrastructure of male accessory glands of Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae).

The ultrastructure of the male accessory glands of the blow fly, Chrysomya megacephala (Fabricius), was presented using light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). A pair of accessory glands was separated at opposite sites. Morphometric results using LM yield evidenced no significant difference in the median of either length or width of the left and right glands. A significant increment in both length and width was seen to plateau between three to six days. SEM observation showed that the surface of the glands revealed a faint irregular groove pattern throughout, and it was occasionally penetrated by tracheoles. Each gland was a slender, elongated sac-like tubule having apical rounded ends, with a slight constriction at the sub-apical part of the gland being observed occasionally. TEM analyses of three-day-old males showed that the glands consisted of external capsular cells with a basement membrane underneath, glandular cells, and gland lumen. The capsular cell was flat and contained a nucleus with electron dense material in the nuclear envelope. The glandular cell, appearing as columnar, consisted of a vacuolated component that contained a large oval nucleus centrally or sub-basally located, with dense mitochondria, numerous rough endoplasmic reticulum, and secretory vesicles containing electron-lucent materials. In the gland lumen, the cross-section through the middle portion revealed dense secretory materials, characterized by electron-dense materials. Some sections revealed a large lumen where secretion accumulates within the delicate sac. The seven-day-old glands exhibited a remarkable change in the lumen, where the whole space contained a large amount of secretory materials, with the electron-dense materials being characterized as similar to those observed in three-day-old glands. About four prominent types of secretions were observed on the basis of difference in electron-density.

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 approximately 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.

Ommatidia of blow fly, house fly, and flesh fly: implication of their vision efficiency.

This work aims to elucidate the number of ommatidia or facets (the outwardly visible units of each ommatidium) for compound eyes in blow flies [Chrysomya megacephala (F.), Chrysomya rufifacies (Macquart), Chrysomya nigripes (Aubertin), Lucilia cuprina (Wiedemann)], house flies (Musca domestica L.), and flesh flies (Liosarcophaga dux Thomson) by manual counts of the corneal spreads. The head of the fly in each species was soaked in 20% potassium hydroxide solution at room temperature for 7 days, and the clear compound eye was dissected into six small parts, each of which was placed onto a slide and flattened using a coverslip. Images of each part were obtained using a microscope connected to a computer. The printed images of each part were magnified, and the total number of ommatidia per eye was manually counted. For males, the mean number of ommatidia was statistically different among all flies examined: L. dux (6,032) > C. rufifacies (5,356) > C. nigripes (4,798) > C. megacephala (4,376) > L. cuprina (3,665) > M. domestica (3,484). Likewise, the mean number of facets in females was statistically different: L. dux (6,086) > C. megacephala (5,641) > C. rufifacies (5,208) > C. nigripes (4,774) > L. cuprina (3,608) > M. domestica (3433). Scanning electron microscopy analysis of adult flies revealed the sexual dimorphism in the compound eye. Male C. megacephala had large ommatidia in the upper two thirds part and small ommatidia in the lower one third part, whereas only small ommatidia were detected in females. Dense postulate appearance was detected in the external surface of the corneal lens of the ommatidia of C. megacephala, C. rufifacies, and C. nigripes, while a mix of dense postulate appearance and variable groove array length was detected in L. cuprina and M. domestica. The probable functions of ommatidia are discussed with reference to other literature.

Fine structure of the reproductive system of Chrysomya megacephala (Diptera: Calliphoridae): the external sexual organ.

The fine structure of the reproductive system of Chrysomya megacephala (Fabricius), a blow fly species of medical importance, was studied using scanning electron microscopy (SEM) to contribute detailed morphological information on the external sexual organs or genitalia of this species. Analysis of the male genitalia or hypopygium revealed that the epandrium is a broad organ resembling a crescent shape, while the ejaculatory apodeme and aedeagal apodeme show similarity in their lengths. The cercus is significantly longer than the surstylus with its apical end being more or less rounded. Dense patches of long bristles, morphologically similar to the sensilla chaetica and sensilla trichodea, were found along the lower half of the cercus. The surstylus has a stout triangular shape, and the proximal half is greatly endowed with sensilla thought to be sensilla chaetica and sensilla trichodea. The aedeagus per se is prominent and has a clavate shape formed by the base theca and elongated phallus. The vesica of the phallus is a smooth bilobed structure, which curves inward. The juxta and juxta process are armed with many rows of strong spines resembling thorns in their appearance. The harpe is slender, recurved anteriorly, and distally pointed like a sickle. Regarding the female genitalia or ovipositor, SEM observation of the supraanal plate revealed it to be sclerotized and more or less triangular-shaped, lying between a pair of short cerci. Several types of sensilla were found on the supraanal plate, subanal plate and cercus, including the sensilla trichodea, sensilla basiconica, sensilla placodea, and probably, sensilla styloconica. The possible functions of sensilla distributed in particular regions of the fly genitalia are discussed.

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.

Male genitalia of flesh fly Parasarcophaga (Liosarcophaga) dux (Diptera: Sarcophagidae) revealed by scanning electron microscopy.

The male genitalia of flesh fly Parasarcophaga (Liosarcophaga) dux (Diptera: Sarcophagidae) was observed using scanning electron microscopy (SEM). The phallus is a short, broad structure that is formed by a tubular base connected to a trumpet-shaped, anteroventrally expanded ventralia. The juxta projects forward from a broad base, which is connected to an upright long structure, the harpes. The juxta is apically bifurcated. The lateral styli bear three small spines curved inward. The pregonite and postgonite are slightly curved upward apically. The cerci are pointed and curved apically. Sensillae were observed on the surfaces of postgonite, surstylus, epandrium, and cerci.