Wound Myiasis Caused by Sarcophaga (Liopygia) Argyrostoma (Robineau-Desvoidy) (Diptera: Sarcophagidae): Additional Evidences of the Morphological Identification Dilemma and Molecular Investigation.

In Mediterranean countries, Sarcophaga (Liopygia) crassipalpis, Sarcophaga (L.) argyrostoma, and Sarcophaga (L.) cultellata share the same ecological niche and can be responsible of myiasis. In this study, the main morphological characters of a larva found in a hospitalized woman were described and illustrated by light and SEM microscopy and the features discussed. Then, a fragment within the mitochondrial encoded cytochrome c oxidase subunit I (coxI) gene of ~735 bp was amplified and sequenced. The molecular investigation was necessary to confirm the species Sarcophaga (Liopygia) argyrostoma (99% of identity). Our findings showed that morphological descriptions of larvae of three Mediterranean species of Liopygia available in several papers might not be clear enough to allow for comparison and correct identification. Until results of reliable comparative studies of larvae of all three species will be available, the use of molecular tools is crucial, to avoid misleading or incomplete identification, and in particular when a myiasis becomes a legal issue.

Third instar larvae of flesh flies (Diptera: Sarcophagidae) of forensic importance--critical review of characters and key for European species.

Necrophagous Sarcophagidae are among the insects most frequently reported from human corpses. The broad forensic application of flesh flies is restricted by the lack of reliable tools for species identification of larval stages and mass breeding of collected flesh fly larvae to the adult stage, and more recently DNA-based methods are usually recommended for precise species identification. To overcome this situation, the following study was implemented: (1) original larval material was obtained of the European flesh flies of confirmed or potential forensic importance; (2) larval morphology was studied and documented using a combination of standard light microscopy, image-stacking stereomicroscopy and SEM; and (3) larval characters used in previously published keys were critically revised. The taxonomic value of the following characters was considered insignificant: (1) differences in level of sclerotisation of particular parts of the cephaloskeleton, (2) level of sclerotisation of the posterior spiracular peritreme and (3) the shape of posterior spiracular slits. A high taxonomic value was noticed for the general shape of anterior spiracles, pattern of arrangement of their lobes, and distribution and shape of spines/warts on the inter-band area of segments. Two character states-long window in the dorsal cornu of cephaloskeleton and deep spiracular cavity on anal division-are not found in the Miltogramminae and therefore cannot be considered as family-specific for the entire Sarcophagidae. As a comprehensive result of our studies, an identification key is presented for the third instar larvae of European flesh flies of forensic importance. The key is user-friendly and requires no dissections of larvae, as soaking the material in methyl salicylate will allow observation of all diagnostic details of the cephaloskeleton. A simple stereomicroscope (magnification about ×50) is sufficient for the observation of all characters presented in the key. This key may be systematically extended by the addition of species present in adjacent geographical regions.

Sarcophaga (Liosarcophaga) dux (Diptera: Sarcophagidae): A flesh fly species of medical importance.

BACKGROUND: Although tropical climate of Thailand is suitably endowed with biodiversity of insects, flies of medical importance is not well investigated. Using information from literature search, fly survey approach and specialist's experience, we review database of Sarcophaga (Liosarcophaga) dux Thomson (Diptera: Sarcophagidae), one of the priorities flesh fly species of medical importance in Thailand. RESULTS: This review deals with morphology, bionomics and medical involvement. Important morphological characteristics of egg, larva, puparia and adult were highlighted with illustration and/or micrographs. Search pertaining to molecular analysis used for fly identification and developmental rate of larvae were included. Medical involvement of larvae was not only myiasis-producing agent in humans and animals, but associated with human death investigations. CONCLUSIONS: This information will enable us to accurate identify this species and to emphasis the increase medically important scene in Thailand.

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.

Revision of the Neotropical genus Malacophagomyia (Diptera: Sarcophagidae) with description of a new species.

The small Neotropical genus Malacophagomyia Lopes is revised. Two previously recorded species from tropical South America are redescribed: M. filamenta (Dodge) and M. kesselringi Kano & Lopes. One new species is described, M. rivadavia sp. nov., from temperate southern South America. The structures of the male genitalia of the species of this genus are compared, and some female genitalic structures of M. filamenta are reinterpreted. Diagnostic characters to recognize the three species are given. Additionally, a key to described species of Malacophagomyia is presented.

Sarcophaga (Hoa)flexuosa Ho (Diptera: Sarcophagidae): association of sexes using morphological and molecular approaches, and a redefinition of Hoa Rohdendorf.

Sarcophaga flexuosa Ho, 1934 is the type species of the currently monotypic genus-group taxon Hoa Rohdendorf, 1937, which here is considered a subgenus of Sarcophaga Meigen, 1826. Using DNA sequences of mitochondrial genes (cytochrome oxidase I and cytochrome b), we positively identify, for the first time, the female of S. (H.)flexuosa. The female is described, the male is redescribed, and both sexes are illustrated using a combination of line drawings, photographs and scanning electron microscopy images. It is argued from the morphology of the male terminalia that Sarcophagaflexuosa is the sister taxon of Sarcophaga basiseta Baranov, 1931, and the subgenus Hoa is redefined to include both of these species.

Description of three new species of Dexosarcophaga (Diptera, Sarcophagidae) from Brazil with new information on synonyms and filling gaps in species distribution.

Three new species of DexosarcophagaTownsend, 1917 are described based on male specimens collected in Brazil, Dexosarcophaga sinoisp. nov., Dexosarcophaga autisferasp. nov., and Dexosarcophaga clavissp. nov. Male morphology is represented by photographs and detailed illustrations of terminalia. Dexosarcophaga carvalhoi (Lopes, 1980), Dexosarcophaga globulosaLopes, 1946, Dexosarcophaga limitata (Lopes, 1975), Dexosarcophaga paulistana (Lopes, 1982b), and Dexosarcophaga petraSantos, Pape and Mello-Patiu 2022 are recorded for the first time in Argentina. Also, Dexosarcophaga lenkoiLopes, 1968, Dexosarcophaga montana (Lopes, 1975), and Dexosarcophaga transitaTownsend, 1917 have their distribution expanded with new records. Dexosarcophaga transita is considered a senior synonym of Dexosarcophaga chaetosaBlanchard, 1939syn. nov., Dexosarcophaga itaquaDodge, 1966syn. nov., and Dexosarcophaga lopesiDodge, 1968syn. nov. With the addition of the new species and the new synonymies proposed, Dexosarcophaga now contains 58 species, of which 10 are recorded in Argentina and 35 in Brazil.

A new Sarcophaginae (Diptera: Sarcophagidae) from Brazil: the peculiar colour of Petrolina bifasciata gen. nov., sp. nov., from the Caatinga xeric shrubland of northeastern Brazil.

A new genus and species, Petrolina bifasciata gen. nov., sp. nov. (Diptera: Sarcophagidae, Sarcophaginae), with only two black stripes dorsally on thorax, is described from both sexes. The type material was collected in fragments of seasonally dry tropical forest in the state of Pernambuco, northeastern Brazil. The new genus and species is morphologically similar to genera Rafaelia Townsend and Malacophagula Bequaert. However, the atypical two thoracic black stripes and phallic morphology give evidence to the proposal of a new genus in Sarcophaginae.

Two species of Microcerella (Diptera: Sarcophagidae) found in highland arid landscapes of Argentina, during forensic studies.

Two species of Microcerella Macquart were obtained from pig and llama corpses during a decomposition field experiment in highlands desert landscapes of Catamarca, Argentina. We sampled four pig and two llama carcasses. Microcerella antofagastensis sp. nov. is described, and M. quimaliensis (Lopes, 1982) is redescribed. Illustrations of male and female terminalia of both species are provided and should allow a correct identification. Notes on the biology of both species are given.

A review of morphological characters for the identification of three common European species of Sarcophaga s. str. (Diptera: Sarcophagidae), with an emphasis on female terminalia.

The subgenus Sarcophaga Meigen, 1824 (s. str.) currently comprises over 30 species distributed in the West Palearctic Region, the identification of which is normally based on characters of the male terminalia. Females of the three closely-related species Sarcophaga (Sarcophaga) carnaria (Linnaeus, 1758), S. (S.) subvicina Rohdendorf, 1937 and S. (S.) variegata (Scopoli, 1763), which are especially widespread and abundant in NW Europe, are considered morphologically indistinguishable by most authors. However, a few authors have proposed keys to separate females of these three species based on external and internal characters of the terminalia. Following a preliminary molecular identification using DNA barcode sequences (COI, cytochrome c oxidase subunit I), we herein revise the morphological characters used to differentiate female S. carnaria, S. subvicina and S. variegata in existing identification keys as well as search for additional diagnostic characters. Our results suggest that only one previously-proposed female character, namely the length to width ratio of abdominal sternite 7, can be used to separate S. subvicina from the other two species (Mann-Whitney U test: p < 0.0001), at least in a majority of cases. Other characters, such as the degree of sclerotisation and setation of tergite 8, show a high degree of overlap that does not allow to reliably separate females of these three species. Nevertheless, we propose a combination of characters that should allow the separation of female S. carnaria from female S. variegata in most cases. An additional analysis of males of the same species showed that the distribution of pruinosity and setation on syntergosternite 7+8, a character mentioned in a previously-published key, is also not reliable for identification.

Gastrointestinal myiasis by larvae of Sarcophaga sp. and Oestrus sp. in Egypt: report of cases, and endoscopical and morphological studies.

Four cases of gastric or intestinal myiasis are reported. The cases contain 2 males (1 child 10 years old, and 1 adult 40 years old) and 2 females (1 girl 18 years old, and 1 adult 50 years old) from Minia Governorate, Southern Egypt. Three of them, including cases no. 1, 3, and 4, were gastric myiasis, and complained of offensive hematemesis of bright red blood. Minute moving worms, larvae of the fly, were found in the vomitus. On the other hand, case no. 2 had intestinal myiasis, and complained of abdominal distention, nausea, vomiting, and diarrhea. The stool of case 2 was mixed with blood, and minute moving worms were observed in the stool. Endoscopy was performed to explore any pathological changes in the stomach of the patients. The larvae were collected and studied macroscopically, microscopically, and us-ing a scanning electron microscope (SEM) to identify their species. Three different types of larvae were identified. The larvae isolated from case 1 were diagnosed as the second stage larvae of Sarcophaga species, and the larvae isolated from case 2 were the third stage larvae of Sarcophaga species. On the other hand, the larvae isolated from cases 3 and 4 were diagnosed as the third stage larvae of Oestrus species.

Description of Two New Species of Dexosarcophaga (Diptera: Sarcophagidae) From the Coastal Environments of the Brazilian Amazon.

Two new species of Dexosarcophaga Townsend (Diptera: Sarcophagidae) are described, both from the coastal zone of the Brazilian Amazon: Dexosarcophaga salgada sp. nov. and Dexosarcophaga klycyae sp. nov.

First instar larvae of endemic Australian Miltogramminae (Diptera: Sarcophagidae).

The first instar larva of a species of the Australian endemic genus Aenigmetopia Malloch is described for the first time, along with the first instar larvae of three other Australian species representing the genera Amobia Robineau-Desvoidy and Protomiltogramma Townsend. Larval morphology was analysed using a combination of light microscopy, confocal laser scanning microscopy and scanning electron microscopy. The following morphological structures are documented: pseudocephalon, antennal complex, maxillary palpus, facial mask, modifications of thoracic and abdominal segments, anal region, spiracular field, posterior spiracles and details of the cephaloskeleton. Substantial morphological differences are observed between the three genera, most notably in the labrum and mouthhooks of the cephaloskeleton, sensory organs of the pseudocephalon, spinulation, sculpture of the integument and form of the spiracular field. The first instar larval morphology of Aenigmetopia amissa Johnston, Wallman, Szpila & Pape corroborates the close phylogenetic affinity of Aenigmetopia Malloch with Metopia Meigen, inferred from recent molecular analysis. The larval morphology of Amobia auriceps (Baranov), Protomiltogramma cincta Townsend and Protomiltogramma plebeia Malloch is mostly congruent with the morphology of Palaearctic representatives of both genera.

Random forests for predicting species identity of forensically important blow flies (Diptera: Calliphoridae) and flesh flies (Diptera: Sarcophagidae) using geometric morphometric data: Proof of concept.

Wing shape variation has been shown to be useful for delineating forensically important fly species in two Diptera families: Calliphoridae and Sarcophagidae. Compared to DNA-based identification, the cost of geometric morphometric data acquisition and analysis is relatively much lower because the tools required are basic, and stable softwares are available. However, to date, an explicit demonstration of using wing geometric morphometric data for species identity prediction in these two families remains lacking. Here, geometric morphometric data from 19 homologous landmarks on the left wing of males from seven species of Calliphoridae (n = 55), and eight species of Sarcophagidae (n = 40) were obtained and processed using Generalized Procrustes Analysis. Allometric effect was removed by regressing centroid size (in log10 ) against the Procrustes coordinates. Subsequently, principal component analysis of the allometry-adjusted Procrustes variables was done, with the first 15 principal components used to train a random forests model for species prediction. Using a real test sample consisting of 33 male fly specimens collected around a human corpse at a crime scene, the estimated percentage of concordance between species identities predicted using the random forests model and those inferred using DNA-based identification was about 80.6% (approximate 95% confidence interval = [68.9%, 92.2%]). In contrast, baseline concordance using naive majority class prediction was 36.4%. The results provide proof of concept that geometric morphometric data has good potential to complement morphological and DNA-based identification of blow flies and flesh flies in forensic work.

New carrion-visiting flesh flies (Diptera: Sarcophagidae) from tropical dry forests of Colombia and their phylogenetic affinities.

Many flesh flies of the subfamily Sarcophaginae are very competitive carrion visitors and breeders, and several are synanthropic species of medical and forensic importance. The knowledge of these taxa is still limited, which is more noticeable in sub-sampled ecosystems such as the tropical dry forests of Colombia. Four new species, Blaesoxipha (Gigantotheca) wajiira sp. nov., Lepidodexia deborarangoa sp. nov., Oxysarcodexia luriza sp. nov. and Udamopyga iku sp. nov., are described from tropical dry forests in the Colombian departments of Atlántico, Bolívar, Cesar and La Guajira, all collected using Van Someren-Rydon traps baited separately with fermented fruit, decomposing fish and human feces. The species B. (G.) wajiira sp. nov. is probably synanthropic, while the remaining three new species avoid habitats in and around built-up areas. A phylogenetic parsimony analysis based on 222 morphological characters across 121 taxa of flesh flies recovers each new species nested within their respective genus. The subgeneric affiliation of L. deborarangoa sp. nov. within the large and diverse genus Lepidodexia (sensu lato) is inconclusive, and potential issues of relying solely on external morphological characters for generic and subgeneric classifications are highlighted. The species Lepidodexia (Notochaeta) woodi (Lopes), Oxyvinia wicharti (Lopes) and Sarcofahrtiopsis spinetta Mulieri & Dufek are new records for Colombia. COI sequence data are provided for U. iku sp. nov., L. (N.) woodi and O. wicharti.

Wing morphometric analysis of forensically important flesh flies (Diptera: Sarcophagidae) in Thailand.

Flesh flies are insects of forensic importance as the larvae associated with human remains can be used to estimate a minimum post-mortem interval (PMImin) in most cases. And, because life-history traits can vary across species, correct identification is a mandatory step before being used as evidence. Adult flesh flies are extremely similar in general appearance, which causes difficulty in species identification as it is largely based on the morphology of the male genitalia; this also makes it difficult to identify females. Currently, landmark-based geometric morphometric analysis of insect wings has proven to be a valuable tool for species identification. Herein, we applied wing morphometric analysis of 524 flesh fly specimens comprising 12 species from Thailand. The right wing of each specimen was removed, mounted on a microscope slide, photographed, and digitized using 18 landmarks. Wing shape variation among genera and species were analyzed using canonical variate analysis, while wing shape variation between sexes of each species was analyzed using discriminant function analysis. A cross-validation test was used to evaluate the reliability of classification. Results of this study demonstrate wing shape can be used to separate genera and species, and distinguish between sexes of the same species, with high reliability. Therefore, the landmark-based geometric morphometric analysis of wings is a useful additional method for species and sex discrimination of flesh flies.

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.

Taxonomic Revision of Lipoptilocnema (Diptera: Sarcophagidae), With Notes on Natural History and Forensic Importance of Its Species.

Lipoptilocnema Townsend is a small genus of Neotropical Sarcophaginae with a distinctive genitalic morphology. This genus is revised based on the examination of the type series and large numbers of specimens of the seven previously known species, plus three new ones herein described, one from Argentina (L. delfinado Mulieri and Mello-Patiu, sp. nov.), and two from Brazil (L. savana Mulieri and Mello-Patiu, sp. nov. and L. tibanae Mulieri and Mello-Patiu, sp. nov.). All species are described or redescribed and illustrated. Distribution maps and a key for male identification are provided. The taxonomic position of this genus is reviewed and the interpretation of phallic structures is discussed. Notes on the natural history of Lipoptilocnema species are provided, and their potential importance as PMI indicators is highlighted, including the first record of Lipoptilocnema reared from a dead human body.

Retrocitomyia Lopes, 1982 (Diptera: Sarcophagidae): new species, new records, key to males, and an updated catalog.

Prior to this work, Retrocitomyia Lopes (Diptera: Sarcophagidae) included ten recognized species distributed exclusively in the New World. We here add a new species to the genus, Retrocitomyia sisbiota sp. nov. from the Central-West Region of Brazil. Four other species of Retrocitomyia are recorded from this region for the first time, redescribed, compared, and illustrated, with emphasis on the male terminalia. Retrocitomyia paraguayensis Lopes is recorded from Brazil for the first time, R. fluminensis Lopes and R. retrocita (Hall) are newly recorded from the Brazilian states of Mato Grosso and Mato Grosso do Sul, and R. mizuguchiana Tibana & Xerez is newly recorded from the state of Mato Grosso do Sul. New generic diagnostic characters are added and the interpretation of some structures of the male terminalia is discussed. An updated catalog of all Retrocitomyia species and an identification key to the males of these species are also provided.

Catalog of the Paramacronychiinae of China (Diptera: Sarcophagidae).

An updated taxonomic catalog of all 32 species of the subfamily Paramacronychiinae so far known to occur in China is presented. The catalog includes data on type locality, references, distribution and information on type material for nominal species with a Chinese type locality. Additional specimens deposited in the Institute of Zoology, Chinese Academy of Sciences and the Museum of Beijing Forestry University were examined. The genus Wohlfahrtiodes Villeneuve, 1910 and the species Wohlfahrtiodes marzinowskyi Rohdendorf, 1962 are new records from China. Two new synonyms are proposed: Wohlfahrtia Brauer & Bergenstamm, 1889 = Sinotibetomyia Xue in Xue & Fei, 2011, syn. nov., and Wohlfahrtia atra Aldrich, 1926 = Sinotibetomyia curvifemura Xue & Fei, 2011, syn. nov.