Two New Species of Oxysarcodexia Townsend (Diptera: Sarcophagidae) From the Chaco Forest Ecoregion of South America.

Two new Neotropical species of the genus Oxysarcodexia Townsend (Diptera: Sarcophagidae) from Chaco ecoregion of Argentina are described, O. berthet, sp. nov. and O. ibera, sp. nov. Both species were recovered from localities in the eastern part of the Chaco forest (Humid Chaco). The descriptions are based on the male external morphology and terminalia. Photographs and SEM images are provided to aid in the identification of the species.

The Auditory System of the Dipteran Parasitoid Emblemasoma auditrix (Sarcophagidae).

Several taxa of insects evolved a tympanate ear at different body positions, whereby the ear is composed of common parts: a scolopidial sense organ, a tracheal air space, and a tympanal membrane. Here, we analyzed the anatomy and physiology of the ear at the ventral prothorax of the sarcophagid fly, Emblemasoma auditrix (Soper). We used micro-computed tomography to analyze the ear and its tracheal air space in relation to the body morphology. Both tympana are separated by a small cuticular bridge, face in the same frontal direction, and are backed by a single tracheal enlargement. This enlargement is connected to the anterior spiracles at the dorsofrontal thorax and is continuous with the tracheal network in the thorax and in the abdomen. Analyses of responses of auditory afferents and interneurons show that the ear is broadly tuned, with a sensitivity peak at 5 kHz. Single-cell recordings of auditory interneurons indicate a frequency- and intensity-dependent tuning, whereby some neurons react best to 9 kHz, the peak frequency of the host's calling song. The results are compared to the convergently evolved ear in Tachinidae (Diptera).

Morphology of immature stages of flesh flies, Boettcherisca nathani and Lioproctia pattoni (Diptera: Sarcophagidae).

The flesh flies are medically-important because the larvae found in the human corpses can provide evidence in forensic investigations through larva identification and their developmental rate. Firstly, we thoroughly described the larval morphology of Boettcherisca nathani and Lioproctia pattoni, using scanning electron microscopy (SEM). The third instar of the two species differed markedly in two characters: (1) spines between the prothorax and mesothorax-B. nathani has more or less slender triangular spines, with those at the posterior region more slender than the anterior region; whereas L. pattoni has stout triangular spines with one or two tips anteriorly, with smaller and tapered triangular shape, grouped two to four laterally in the posterior end, and (2) morphology of the peristigmatic tufts at the posterior spiracle-B. nathani has extensively branched long, fine hairs, whereas tufts in L. pattoni have moderately branched long, fine hairs. The anterior spiracle displayed similarity; B. nathani has two irregular rows of 21-27 papillae, while L. pattoni has a single irregular row of 20-28 papillae. Secondly, we use light microscopy to compare morphology of the third instar of the two species and additional three species, i.e., Bercaea africa, Parasarcophaga dux and Liopygia ruficornis. Particular attention was paid to the features of anterior spiracle, spines between prothorax and mesothorax and posterior spiracle. These results are useful in species identification and estimation of age of larvae found associated with corpses.

Ultrastructural Morphology of the Antenna and Maxillary Palp of Sarcophaga tibialis (Diptera: Sarcophagidae).

A scanning electron microscopy investigation of the antenna and maxillary palp of the adult of Sarcophaga tibialis Macquart (Diptera: Sarcophagidae), a species of medical, veterinary, and forensic relevance, is presented for the first time. Adults of both sexes used in this study were obtained from larvae collected in a case of traumatic myiasis in a domestic cat in northern Italy. The antenna of S. tibialis is that typical of cyclorrhaphan Diptera, consisting of three segments: the scape, the pedicel, and the postpedicel, bearing the arista. The scape is covered by microtrichia and has a row of long chaetic sensilla. The pedicel is also covered by microtrichia and has three types of chaetic sensilla and a cluster of setiferous plaques. Trichoid, styloconic, clavate, and basiconic sensilla are distributed among the microtrichia on the postpedicel. Invaginated basiconic-like sensilla and olfactory pits are also present, the latter ones more numerous in the female. Our results are compared with those obtained for other calyptrate flies, mainly in the family Sarcophagidae. The data obtained may represent a basis for electrophysiological studies on the sensorial activity of the species related to the search for food sources, mates, and suitable larviposition sites, and for comparative morphological studies with other Diptera.

Ultrastructure analysis of the immature stages of Ravinia belforti (Diptera: Sarcophagidae), a species of medical-veterinary and forensic importance, by scanning electron microscopy.

The postmortem interval is related to the age of immature species of flies found on corpses and can be estimated using data available in the literature on the biology of the species. The flesh fly Ravinia belforti is a carrier of enteric pathogens that can affect human and animal health as well as being of forensic importance. As the morphology of many immature Sarcophagidae is unknown, these immature forms must be collected and characterized after the emergence of the adult male. Here we describe and analyze the morphological characteristics of the larvae stages L1, L2, L3 and the puparium of R. belforti by scanning electron microscopy (SEM). Ten specimens of each stage were analyzed. Larvae of R. belforti follow the typical muscoid vermiform pattern with 12 segments. The anterior region is pointed, while the posterior region is thicker. The spines of the cephalic collar are flattened and with double, triple or quadruple points, different from the spines along the body that only have a single point. In L2, the anterior spiracle is present with a varying number of papillae (16-22), differing from other species. The posterior spiracles are located within the peritreme. The spiracular cavity is internalized in the posterior region, following the pattern that differs Sarcophagidae from other families. L3 features more visible and developed spines around the cephalic collar, getting thicker and denser near to the first thoracic segment. Puparium is similar to other species of Sarcophagidae. This paper presents important data on this family which has both health and forensic importance. Furthermore, R. belforti shows significant differences from other species of Sarcophagidae.

Sarcophaga (Liosarcophaga) tibialis Macquart 1851 (Diptera: Sarcophagidae): micromorphology of preimaginal stages of a fly of medical and veterinary interest.

Sarcophagids are a large family of Diptera, with a worldwide distribution. They are related to decomposing organic matter and are very interesting for health science and in forensic cases since many species produce myiasis and occur in human corpses. This family is considered difficult to study, particularly with regard to their immature stages, to which little attention has been paid. Genus Sarcophaga Meigen, 1826 is composed of species of very similar morphology, making very difficult to distinguish. Knowledge of the immature stages of this genus is important because such stages occupy the greater part of the life cycle, so that establishing a basis for their identification will increase their usefulness in systematic and applied sciences. This contribution presents a detailed study of the morphological features, both external and internal, of the preimaginal stages of Sarcophaga (Liosarcophaga) tibialis Macquart, 1851, providing a taxonomical context for the correct identification of Liosarcophaga species of forensic interest in the Iberian Peninsula. Both light and scanning electron microscopy were applied. Complete descriptions of every stage are provided and illustrated, and their usefulness for species comparison, taking into account our uneven knowledge of morphologically immature stages of this subgenus, is indicated. Features of the cephalopharyngeal skeleton, such as the shape of the mouth hook and the intermediate and basal sclerites, and external morphology, such as the pattern of spinose band and anterior and posterior spiracles, proved useful for separating species. Finally, tentative identification keys based on light microscopy observation to distinguish S. (L.) tibialis from other species of forensic interest belonging to Liosarcophaga subgenus are proposed for every immature stage.

Micromorphology of immature stages of Sarcophaga (Liopygia) cultellata Pandellé, 1896 (Diptera: Sarcophagidae), a forensically important fly.

The Sarcophagidae are one of the most numerous groups of Diptera in the world, consisting of many species of forensic interest, whose immature stages are useful in the estimation of postmortem interval. The immature stages of some species of forensic importance still remain unknown or undescribed, like in the case of Sarcophaga (Liopygia) cultellata Pandellé, 1986, a species restricted to the Iberian Peninsula, south of France and north of Italy, which shares a ecological niche with species of the same subgenus, e.g., Sarcophaga (Liopygia) argyrostoma (Robineau-Desvoidy, 1830) and Sarcophaga (Liopygia) crassipalpis Macquart, 1839, making it necessary to lay the groundwork for a proper specific differentiation before it can be successfully applied in forensic practice. This study provides the first micromorphological description of all preimaginal stages of S. (L.) cultellata using light microscopy and scanning electron microscopy (SEM), the results of which allow the morphology of the main features to be followed during the immature life cycle. We propose a combination of features for distinguishing Liopygia from other sarcophagid subgenera, based on the current level of morphological knowledge of immature stages. S. (L.) cultellata can be differentiated from S. (L.) argyrostoma and S. (L.) crassipalpis in every immature stage by both light microscopy and SEM. The presence of tegumental warts and a fan-shaped anterior spiracle with a single row of 15-18 respiratory papillae allow distinguishing the third instar larvae of S. (L.) cultellata from other Sarcophaga species described hitherto by SEM. Identification keys based on light microscopy observations are provided, covering all the immature stages of Liopygia subgenus occurring in the Iberian Peninsula.

Morphology of puparia of flesh flies in Thailand.

Puparia of five flesh fly species were investigated for forensic study. Boettcherisca nathani (Lopes, 1961), Boettcherisca peregrina (Robineau-Desvoidy, 1830), Lioproctia pattoni (Senior-White, 1924), Liopygia ruficornis (Fabricius, 1794) and Parasarcophaga (Liosarcophaga) dux (Thomson, 1869) were examined with a scanning electron microscopy (SEM). Differences between species were found in the number and arrangement of papillae in the anterior spiracle, the shape of intersegmental spines between the prothorax and mesothorax and the pattern of spiracular tufts at the posterior spiracle. The anterior spiracle of B. nathani had two rows, comprising 21-27 papillae; while those of B. peregrina and L. pattoni had one or two irregular rows with 24-26 and 20-28 papillae, respectively. Anterior spiracle of L. ruficornis and P. dux had one row of 10-15 papillae. Intersegmental spines between the prothorax and mesothorax and pattern of spiracular tufts at the posterior spiracle are morphologically different. L. ruficornis and P. dux puparia are similar, but the position of the interslit plate between the inner and middle spiracular slits was found to be an important attribute to separate both species. Morphometric analysis on the length and width of puparia of these species revealed statistically different among them. The key for identifying puparia of forensically important flesh flies has been provided.

Morphology of the first instar larva of obligatory traumatic myiasis agents (Diptera: Calliphoridae, Sarcophagidae).

There are only three fly species that are obligate agents of traumatic myiasis of humans and livestock: a single species of flesh fly, Wohlfahrtia magnifica (Sarcophagidae), and two species of blow flies, Chrysomya bezziana and Cochliomyia hominivorax (Calliphoridae). The morphology of their first instar larvae is thoroughly and consistently documented here with light microscopy photographs and scanning electron microscopy micrographs. The following morphological structures are documented: pseudocephalon, antennal complex, maxillary palpus, oral ridges, thoracic and abdominal spinulation, spiracular field, posterior spiracles and cephaloskeleton. New diagnostic features drawn from the cephaloskeleton and the spinulation of abdominal segments, including the anal pad, are discovered and extensively described. Earlier descriptions in the literature are revisited, and major discrepancies between these and the results of the current study are discussed. The present results allow clarification, correction and, especially, complementation of information provided by earlier authors. The relatively distant taxonomic position of all three species is evidence that obligatory myiasis has arisen independently, and the extensively similar morphology in the first instar larvae of Chrysomya bezziana, Cochliomyia hominivorax and W. magnifica in comparison to necrophagous species, especially the enhancement of the anterior part of the cephaloskeleton and the segmental spinulation, is therefore best interpreted as homoplasic adaptations to a life strategy as obligate vertebrate parasites. An identification key for first instar larvae of all obligatory traumatic myiasis agents of mammals is provided.

Useless hearing in male Emblemasoma auditrix (Diptera, Sarcophagidae)--a case of intralocus sexual conflict during evolution of a complex sense organ?

Sensory modalities typically are important for both sexes, although sex-specific functional adaptations may occur frequently. This is true for hearing as well. Consequently, distinct behavioural functions were identified for the different insect hearing systems. Here we describe a first case, where a trait of an evolutionary novelty and a highly specialized hearing organ is adaptive in only one sex. The main function of hearing of the parasitoid fly Emblemasoma auditrix is to locate the host, males of the cicada species Okanagana rimosa, by their calling song. This task is performed by female flies, which deposit larvae into the host. We show that male E. auditrix possess a hearing sense as well. The morphology of the tympanal organ of male E. auditrix is rather similar to the female ear, which is 8% broader than the male ear. In both sexes the physiological hearing threshold is tuned to 5 kHz. Behavioural tests show that males are able to orient towards the host calling song, although phonotaxis often is incomplete. However, despite extensive observations in the field and substantial knowledge of the biology of E. auditrix, no potentially adaptive function of the male auditory sense has been identified. This unique hearing system might represent an intralocus sexual conflict, as the complex sense organ and the behavioural relevant neuronal network is adaptive for only one sex. The correlated evolution of the sense organ in both sexes might impose substantial constraints on the sensory properties of the ear. Similar constraints, although hidden, might also apply to other sensory systems in which behavioural functions differ between sexes.

Structural and Genetic Investigation of the Egg and First-Instar Larva of an Egg-Laying Population of Blaesoxipha plinthopyga (Diptera: Sarcophagidae), a Species of Forensic Importance.

Flies in the family Sarcophagidae incubate their eggs and are known to be ovoviviparous (i.e., ovolarviparous), but a laboratory-maintained colony of Blaesoxipha plinthopyga (Wiedemann) deposited clutches of viable eggs over 10 generations. A description of the egg and first-instar larva of this species is provided along with genetic data (genome size and cytochrome oxidase I sequences). The egg is similar to previously described eggs of other Sarcophagidae but differs in the configuration of the micropyle. In the first-instar larva, the oral ridges are much more developed than has been described for other species. B. plinthopyga has forensic importance, and the present descriptive information is critical for proper case management.

Ultrastructure of immature stages of Peckia (Euboetcheria) collusor (Diptera: Sarcophagidae).

Peckia (Euboettcheria) collusor (Diptera: Sarcophagidae) is a necrophagous insect and represents an important role in forensic entomology, as it is always collected at animal/human corpses. Immature sarcophagids are extremely similar in morphological aspects and can be found colonizing both fresh and mummified bodies. Flesh fly larvae are reported to cause accidental myiasis. In the Neotropical regions little is known about the ultrastructure of immature stages of Sarcophagidae. Scanning electron microscopy gives detailed information on the morphological characters, which can help identify the immature forms of the flies. The aim of this study is to identify larvae and puparia of P. (E.) collusor using this methodology. The integument with net-like patches and the shape and disposition of intersegmental spines of the larval instars can be used to distinguish this flesh fly larva. The number of spiracular ramifications is variable at larval instars. The puparia are very similar to third instar larvae, except for the cephalic structures that are retracted. In conclusion, scanning electron microscopy could help entomologists to identify the immature forms of P. (E.) collusor in forensic investigations and consequently serves as a tool in crime scene investigations.

Malpighian tubules of adult flesh fly, Sarcophaga ruficornis Fab. (Diptera: Sarcophagidae): an ultrastructural study.

The Malpighian tubules of adult flesh fly, Sarcophaga ruficornis consist of principal and stellate cells. The principal cells reveal all the characteristics of transporting epithelia with well developed deep basal membrane infoldings forming a complex of interconnecting labyrinth of canaliculi and luminal microvilli, both of which are associated with mitochondria. The central cytoplasm of the cells contains a well developed nucleus, clear vacuoles or vacuoles filled with secretory material, mineral concretions or spherocrystals, lysosomes and a network of endoplasmic reticulum. The mineral concretions are also observed in the region of luminal microvilli and in the lumen of the tubule suggesting their extrusion into the lumen by exocytosis. Several formed bodies are also observed in the lumen. Stellate cells are characterized by simple membrane infoldings and luminal microvilli devoid of mitochondria. The cells are separated by septate junctions. The Malpighian tubules are richly supplied by tracheae and muscle fibers.

Ultramorphological characteristics of immature stages of a forensically important fly Parasarcophaga ruficornis (Fabricius) (Diptera: Sarcophagidae).

Ultrastructure of all larval instars and puparium of Parasarcophaga ruficornis, a common flesh fly species in India, is presented using light microscopy and scanning electron microscopy for the first time. The principal diagnostic characters, i.e. the cephalopharyngeal apparatus, the cephalic segment, structure and orientation of spines, pupal respiratory horns, the structures of both anterior and posterior spiracles are illustrated and discussed. Information presented herein can be useful during forensic investigations involving this fly species.