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.

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

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.

Identification of forensically important fly eggs using a potassium permanganate staining technique.

Fly eggs found in corpses can be utilized as entomological evidence in forensic investigations of deaths if the species of fly and the developmental rate at a temperature similar to the death scene are known. The species identification of fly eggs is particularly important, and previously, scanning electron microscope has been used for this purpose. Herein, we report a simple technique, using light microscopy, to identify forensically important eggs of Chrysomya rufifacies (Macquart), Chrysomya megacephala (Fabricius), Chrysomya pacifica Kurahashi, Chrysomya nigripes Aubertin, Aldrichina grahami (Aldrich), Lucilia cuprina (Wiedemann), Musca domestica L. and Megaselia scalaris (Loew). A 1% potassium permanganate solution was used to stain egg surfaces for 1 min, followed by dehydration in 15, 70, and 95%, absolute alcohol (each solution for 1 min) and the eggs were permanently mounted. The characteristics are based on the width of plastron, morphology of plastron area surrounding the micropyle and chorionic sculpturing, with the length of egg being used as supplemental feature.

Antennal sensilla of some forensically important flies in families Calliphoridae, Sarcophagidae and Muscidae.

Antennal sensilla of some forensically important fly species in the families Calliphoridae (Chrysomya megacephala, Chrysomya rufifacies, Chrysomya nigripes and Lucilia cuprina), Sarcophagidae (Parasarcophaga dux) and Muscidae (Musca domestica) were studied using scanning electron microscopy. Five types of sensilla were observed: trichoid, basiconic, coeloconic, styloconic and sensory pit. Only trichoid sensilla are found on the scape of the antenna, while both trichoid and styloconic sensilla are located on the antennal pedicels of all species studied. Basiconic sensilla are the most numerous of the sensilla found on the antennae of both sexes of all fly species studied and are comprised of two subtypes: large and small basiconic sensilla. Coeloconic sensilla are characterized by short pegs, with either grooved or smooth surfaces, that are sunken into deep depressions. No marked difference was observed in the number, morphological structure or distributional pattern of any of the sensilla among the species studied, with the exception of there being more numerous sensory pits detected in female P. dux compared to the other species. The suggested function of each antennal sensillum was based on comparison with results of other investigations on similar sensilla.

Larval morphology of Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) using scanning electron microscopy.

The larval morphology of Chrysomya megacephala (Fabricius) is presented using scanning electron microscopy (SEM). Extreme similarity of this species to Chrysomya rufifacies (Macquart), a species usually found concurrently inhabiting decomposing human corpses in Thailand, is seen only in the first-instar larvae. The relative thickness of the branches of the posterior spiracular hairs in these species could be used to differentiate them in this developmental stage. In contrast, the "hairy" appearance of C. rufifacies allows second- and third-instar larvae to be easily distinguished. Results of this study should help in future endeavors to differentiate C. megacephala from other larvae found in decomposing human corpses in Thailand.

Biogeography and Trypanosoma cruzi infection prevalence of Chagas disease vectors in Texas, USA.

Data were pooled from multiple sources including newly collected triatomine specimens, preserved specimens, government reports, and scientific articles to create a biogeographical profile of triatomine vector species found in Texas. Triatomine specimens were documented in 97 of 254 counties, and Trypanosoma cruzi-infected specimens were reported from 48 counties. Triatomine specimens were distributed in 11 of the 12 ecoregions in Texas, with all but one species found in multiple ecoregions. Of the 241 newly collected specimens, 50.74% were infected with T. cruzi. Triatoma gerstaeckeri was the most frequently collected and most geographically dispersed species followed by T. sanguisuga. Three species, T. gerstaeckeri, T. sanguisuga, and T. lecticularia, were associated with human dwellings, and over half of the new specimens found inside or near houses were infected with T. cruzi. Chagas disease vectors in Texas are widely distributed and have adapted to ecologically diverse settings. The high T. cruzi infection prevalence of specimens found in close proximity to human settings suggests the presence of an active peridomestic Chagas disease transmission cycle.

Detection of Blastocrithidia spp. (Kinetoplastida: Trypanosomatidae) in Chagas disease vectors from Texas, USA.

Organisms highly similar to Blastocrithidia triatomae, a pathogenic parasite of Chagas disease triatomine bug vectors, were detected using polymerase chain reaction amplification and DNA sequence analysis of a segment of the small subunit rRNA gene in 3 of 203 triatomine specimens collected in Texas from June 2005 to October 2006. The parasite was identified in three species of triatomine bugs (Triatoma gerstaeckeri, T. indictiva, and T. neotomae) collected from three distinct geographic locations. Flagellated organisms indistinguishable from Trypanosoma cruzi were observed by direct microscopy in hindgut material of two of the three specimens. Coinfection with T. cruzi and Blastocrithidia was detected by molecular methods in one of the specimens. Parsimony analysis provided strong support for clustering of the new sequences within a Blastocrithidia group, clearly separated from other flagellated protozoans. Confirmation of Blastocrithidia in U.S. triatomine species complicates microscopic diagnosis of T. cruzi due to the morphologic similarity of the parasites.

Behavioral responses of Chrysomya megacephala to natural products.

Knowledge of the behavioral responses of medically important flies to natural stimuli is critical for the development of vector control strategies. We designed, constructed, and operated the dual-choice wind tunnel to investigate the behavioral responses of the blowfly, Chrysomya megacephala (Diptera: Calliphoridae), toward two important extrinsic factors (wind speed and olfactory stimuli) that affect the flight behavior of these flies. The dual-choice wind tunnel appeared as a "T-box," consisting of two stimulus partitions and one median release partition. The correlation between wind speed and fly response showed a marked relationship, with the wind speed set at 0.5 m/s being the optimal speed. Of the 72 natural products tested, 24--all having high protein content--attracted flies at the level of greater than 50% in the rearing cages during preliminary tests, thus allowing further investigation in the dual-choice wind tunnel to further assess these responses. The behavioral response between flies exposed to the screening odors in a rearing cage and those exposed in the dual-choice wind tunnel showed a similar trend. One-day-tainted pork viscera yielded the greatest positive response among natural products tested, attracting greater than 90% of the flies included in the test population. One-day-tainted pork viscera exhibited the highest potency (1.578), exceeding 1-day-tainted beef liver, which is commonly used as the standard bait for blowflies. The usefulness and practical applications of the dual-choice wind tunnel for further investigations are also discussed.

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.

Efficacy of neem extract against the blowfly and housefly.

The blowfly, Chrysomya megacephala (Fabricius), and housefly, Musca domestica Linnaeus, are ubiquitous insects that have the potential to spread a variety of pathogens to humans and livestock. Pest management techniques for populations of these flies are needed. Currently, bioinsecticides, particularly those derived from plant origin, have been increasingly evaluated in controlling populations of medically important insects. In this study, an attempt was made to evaluate the efficacy and biological activity of a commercially available neem extract, containing 0.24% azadirachtin A, against C. megacephala and M. domesitca. Laboratory bioassays were performed using the feeding method of mixing neem solutions with fresh beef, once or multiple times, as food for rearing third instar. The laboratory tests showed that neem products significantly reduced larval and pupal survival, adult emergence, pupal weight, adult wing length, and fecundity on the subsequent generation, in a dose-dependent manner in both species. Efficacy was observed in the first generation and could extend to the second generation. Despite these reductions, reduction in total adult longevity was not evident for larvae fed once on neem solutions with fresh beef, and slightly earlier (approximately 1 week) mortality was observed in both species when they were fed as larvae on multiple doses. Adverse effects of this neem-based product toward M. domestica were slightly greater than those in C. megacephala. These data reinforced the efficacy of neem extract in reduced adult emergence and anti-fecundity in the subsequent generation. However, neem extract induced only low to moderate larval and pupal mortalities.

Fine structure of the alimentary canal of the larval blow fly Chrysomya megacephala (Diptera: Calliphoridae).

Morphology of the alimentary canal of the mature third instar larva of the blow fly, Chrysomya megacephala (F.), was examined using light, scanning, and transmission electron microscopy. Salivary structures consist of a single median deferent duct that bifurcates into efferent ducts connected to paired, tubular salivary glands comprised of closely packed conical-shaped epithelial cells with large nuclei. The crop occurs as a large, swollen diverticulum of the digestive tube and is lined internally with convoluted cuticle (epicuticle and endocuticle). The esophagus is a simple, straight tube internally lined with cuticle and externally encompassed by muscle fibers. The cardia is a bulb-like structure composed of anterior foregut tissue and posterior midgut tissue from which the peritrophic membrane (PM) is produced. The midgut begins within the cardia which is flanked posteriorly by four tubular gastric caeca that are lined internally with four to five layers of cuboidal epithelial cells bearing microvilli. Midgut tissue is lined with simple cuboidal epithelium whose cells are filled with numerous secretory granules and possessed long microvilli facing the lumen. A peritrophic membrane is contained within the midgut lumen. The larval hindgut consists of the pylorus, Malpighian tubules, ileum, colon, rectum, and anus, posteriorly. The pylorus is characterized by a single layer of epithelial cells encircled by a muscular layer and the presence of PM within the lumen. Malpighian tubules each diverge into two tubular structures totalling four long tubules of long chained cuboidal cells bearing microvilli internally. The wall of the ileum is comprised primarily of a monolayer of cuboidal epithelial cells with large oval nuclei and more intense muscular fibers surrounding the periphery. A cuticular layer surrounds the lumen containing the PM. This inner cuticle consists of a thin epicuticle that is electron-dense; whereas, the endocuticle is much thicker but less electron-dense. Myo-epithelial cells are dense in the anal region, where the PM persists.

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.

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 approximately 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.