Succession of medico-legal important flesh flies (Diptera: Sarcophagidae) in the temporal gradient of pig decomposition in the Brazilian Cerrado.

Flies belonging to the Sarcophagidae family play a significant role in forensic investigations by aiding in the estimation of post-mortem interval through the assessment of the developmental time of their immature stages and/or the succession patterns of species on carcasses and cadavers. In this regard, this study aimed to investigate the ecological succession of these flies on pig carcasses within Cerrado of Brazil. The progression of fly succession was examined over a 10-day period using the Threshold Indicator Taxa Analysis (TITAN) approach. Six Z + species (Oxysarcodexia thornax (Walker), Peckia (Sarcodexia) lambens (Wiedemann), Peckia (Euboettcheria) collusor (Curran & Walley), Ravinia belforti (Prado & Fonseca), Tricharaea (Sarcophagula) canuta (Wulp) and Tricharaea (Sarcophagula) occidua (Fabricius)) were identified, with change points ranging from 2.5 to 3 days during the dry season and 2.5 to 5.5 days during the rainy season. Two Z- species (Dexosarcophaga carvalhoi (Lopes) and Peckia (Sarcodexia) tridentata (Hall)) were present, with a change point of 6.5 days during the rainy season. This study provides a continuous analysis of the temporal succession of flies, enabling an investigation into species progression based on their change points and directions (Z + and Z- ), thereby offering methodological advancements to avoid the arbitrary categorization of inherently continuous data.

Diversity response of necrophagous dipteran communities and their functional groups to an anthropization gradient.

The anthropization process exerts a profound effect on ecosystems, causing alterations in biodiversity, habitat structure, and species composition, ultimately disrupting the delicate balance of natural environments. The aim of the present study was to explore the ecological dynamics of necrophagous Sarcophagidae and Calliphoridae flies along an anthropization gradient. This research investigated alpha and beta diversity patterns to unravel the impact of human-induced environmental changes on these insect communities and also assesses the dynamics of functional groups in relation to their impact on medical and forensic fields. Five distinct habitats, ranging from urban to forested areas, were surveyed in two Departments in the Province of Chaco, Argentina. Necrophagous flies were collected using van Someren-Rydon canopy traps across three seasons. Two main functional groups were analyzed: oviparous flies (Calliphoridae) and larviparous flies (Sarcophagidae). Results indicated a significant negative correlation between Sarcophagidae species richness and anthropization, whereas Calliphoridae showed increased abundance in highly anthropized sites. The combined assemblage of Calliphoridae+Sarcophagidae exhibited significant relationships across all community parameters evaluated. Beta diversity analysis revealed turnover as the main process shaping dipteran communities along the anthropization gradient, with spatial species replacement dominating. This underscores the importance of interspecific spatial segregation in dipteran community composition. In conclusion, this study enhances our understanding of the ecological adaptations of necrophagous dipterans to anthropogenic disturbances. The observed shifts in diversity and abundance have implications for forensic investigations and public health, emphasizing the need for nuanced monitoring and conservation strategies. This research contributes valuable insights into the intricate ecological interactions of these insect communities within changing ecosystems.

Immunomodulatory potential of Sarcophaga argyostoma larval hemolymph as a natural alternative to berenil in treating Trypanosoma evansi in vivo.

This study compared effects of diminazene aceturate (berenil), commonly used to treat domestic animals infected with Trypanosoma evansi, with the hemolymph of Sarcophaga argyostoma larva. The hemolymph may be acting as a possible natural alternative to berenil, based on immunomodulation mediated inflammatory response. Inflammatory mediators and histopathological changes in liver, kidney, and spleen of albino mice experimentally infected with T. evansi were studied. Mice were divided into five groups: G1, uninfected, untreated (negative control); G2, T. evansi infected (positive control); G3, infected and treated with berenil; G4, infected and treated with hemolymph; G5, infected and treated with hemolymph 3 days before infection (prophylactic group). Animals in (G4) and (G5) exhibited a significant overall reduction in serum levels of IFN-γ. However, the reduction in TNF-α and IL-6 levels was more limited compared to (G2) and (G3). Notably, an elevation in IL-10 levels was observed compared to animals in other groups. Furthermore, the groups treated with hemolymph demonstrated an alleviation of T. evansi infection in contrast to the other groups. This study highlights that the administration of Sarcophaga argyostoma larval hemolymph at a dosage of 0.5 ml/kg significantly inhibited T. evansi organisms in vivo, showcasing a pronounced trypanocidal effect.

Sarcophagidae (Insecta: Diptera) of Saudi Arabia: new records, an updated checklist and a new species.

As part of a study on the biodiversity of Diptera of south-western Saudi Arabia, a survey of the Diptera fauna of Jazan, Asir and Najran at 10 sites was performed between 20122015 mainly using Malaise traps, sweep nets and baited traps. A total of 21 species of Sarcophagidae are recorded and documented with photographs, seven of which represent new country records for Saudi Arabia. Including species with a reliable previous record from Saudi Arabia, the total number of species of Sarcophagidae known from Saudi Arabia now stands at 51. All species of Sarcophagidae recorded from Saudi Arabia are listed, including distributions and the source where species were first recorded. Material studied as part of the project resulted in 16 new country records for other countries. Alusomyia Villeneuve, 1933 is here synonymized with Hoplacephala Macquart, 1834, syn. nov., argued from the presence of four proclinate orbital bristles, the row of strong parafacial setae close to the anteroventral eye-margin, the large gena and the robust habitus compared to most other miltogrammines. The generic synonomy requires the new combinations Hoplacephala transfuga (Villeneuve, 1933), comb. nov. and Hoplacephala turana (Rohdendorf, 1975), comb. nov. Sarcophaga surcoufi Villeneuve, 1913 is synonymized with Engelisca adhamae Lehrer & Abou-Zied, 2008, syn. nov. An appendix authored by Dr J.C. Deeming contains the description of Sarcophaga dawahi Deeming, sp. nov.

Transcriptomic profiling of different developmental stages reveals parasitic strategies of Wohlfahrtia magnifica, a myiasis-causing flesh fly.

BACKGROUND: Wohlfahrtia magnifica is an obligatory parasite that causes myiasis in several warm-blooded vertebrates. Adult females deposit the first-stage larvae directly onto wounds or natural body orifices (e.g., genitalia) of the host, from where they quickly colonize the host tissue and feed on it for development. The infestation of W. magnifica can lead to health issues, welfare concerns, and substantial economic losses. To date, little is known about the molecular mechanisms of the W. magnifica-causing myiasis. RESULTS: In this study, we collected parasitic-stage larvae of W. magnifica from wounds of naturally infested Bactrian camels, as well as pupae and adult flies reared in vitro from the wound-collected larvae, for investigating the gene expression profiles of the different developmental stages of W. magnifica, with a particular focus on examining gene families closely related to the parasitism of the wound-collected larvae. As key proteins related to the parasite-host interaction, 2049 excretory/secretory (ES) proteins were identified in W. magnifica through the integration of multiple bioinformatics approaches. Functional analysis indicates that these ES proteins are primarily involved in cuticle development, peptidase activity, immune response, and metabolic processes. The global investigation of gene expression at different developmental stages using pairwise comparisons and weighted correlation network analysis (WGCNA) showed that the upregulated genes during second-stage larvae were related to cuticle development, peptidase activity, and RNA transcription and translation; during third-stage larvae to peptidase inhibitor activity and nutrient reservoir activity; during pupae to cell and tissue morphogenesis and cell and tissue development; and during adult flies to signal perception, many of them involved in light perception, and adult behavior, e.g., feeding, mating, and locomotion. Specifically, the expression level analysis of the likely parasitism-related genes in parasitic wound-collected larvae revealed a significant upregulation of 88 peptidase genes (including 47 serine peptidase genes), 110 cuticle protein genes, and 21 heat shock protein (hsp) genes. Interestingly, the expression of 2 antimicrobial peptide (AMP) genes, including 1 defensin and 1 diptericin, was also upregulated in the parasitic larvae. CONCLUSIONS: We identified ES proteins in W. magnifica and investigated their functional distribution. In addition, gene expression profiles at different developmental stages of W. magnifica were examined. Specifically, we focused on gene families closely related to parasitism of wound-collected larvae. These findings shed light on the molecular mechanisms underlying the life cycle of the myiasis-causing fly, especially during the parasitic larval stages, and provide guidance for the development of control measures against W. magnifica.

All insects matter: a review of 160 entomology cases from 1993 to 2007 in Switzerland-part I (Diptera).

Necrophagous Diptera are the most important group of insects used for the purposes of forensic entomology. While the most utilized fly family in this context is the family Calliphoridae, there are several other families that can be of great importance during real-case investigations. This article analyzes the necrophagous flies of all families recorded from 160 real cases in Switzerland between 1993 and 2007. A total of 56 species belonging to 16 families was identified with Calliphoridae being the most dominant family (90.63% of all cases), followed by Muscidae (26.25%), Sarcophagidae (19.38%), Phoridae (14.38%), and Fanniidae (12.50%). For specimens that were difficult to identify morphologically, a new PCR primer has been specifically designed for the amplification of a short, informative COI barcode in degraded museum samples of forensically important Diptera taxa. The richest family in terms of species was the family Muscidae with 16 species. Fannia fuscula (Fallen) and Fannia monilis (Haliday) were recorded from human cadavers for the first time. The study highlights the importance of different fly families in forensic investigation, enhancing our comprehension of their prevalence and dispersion in real cases in Central Europe. The results pave the way for additional exploration, especially regarding the involvement of less frequently observed species in forensic entomology.

First record of Brachymeria amenocles (Hymenoptera: Chalcididae) associated with larvae of Peckia (Peckia) chrysostoma (Diptera: Sarcophagidae) in Brazil.

In this work, we report for the first time the occurrence of the parasitoid wasp Brachymeria amenocles (Walker, 1846) (Hymenoptera: Chalcididae) associated with the larvae of the flesh fly, Peckia (Peckia) chrysostoma (Wiedemann, 1830) (Diptera: Sarcophagidae), in Brazil. This parasitoid species was collected together with the species Brachymeria podagrica (Fabricius, 1787), in the municipality of Rio de Janeiro, geographically located in the Southeast region of Brazil, in larvae of the same host species reared in the same attraction substrate, behaving as solitary parasitoids. A trap containing putrefying sardine was exposed for 48 h, in the upper part of a house, located in Vila Isabel, an urban area. The larvae collected were reared in the laboratory without control of environmental conditions. A total of three adult parasitoid wasps of the species B. podagrica, and nine adult parasitoid wasps of the species B. amenocles emerged from 12 host pupae, resulting in a prevalence of parasitoidism of 4.8% and 14.5%, respectively. Developing parasitoids at the pupal phase were observed in another 34 host pupae dissected.

Molecular identification and functional analysis of chitinase genes reveal their importance in the metamorphosis of Sarcophaga peregrina (Diptera: Sarcophagidae).

Chitinases play a crucial role in insect metamorphosis by facilitating chitin degradation. Sarcophaga peregrina (Robineau-Desvoidy, 1830) (Diptera: Sarcophagidae) is a typical holometabolous insect and an important hygiene pest that causes myiasis in humans and other mammals and acts as a vector for various parasitic agents, including bacteria, viruses, and parasites. Enhancing the understanding of the metamorphosis in this species has significance for vector control. In this study, we identified a total of 12 chitinase genes in S. peregrina using bioinformatic analysis methods. Based on transcriptome data, SpIDGF2 and SpCht10 were selected for further functional investigation. The down-regulation of these genes by RNA interference led to developmental delays, disruptions in molting, and differences in cuticle composition during the pupal stage. These findings underscore the pivotal role of chitinase genes in the metamorphic process and offer valuable insights for effective control strategies.

Effects of heat tolerance on the gut microbiota of Sarcophaga peregrina (Diptera: Sarcophagidae) and impacts on the life history traits.

BACKGROUND: Heat tolerance is a distinct abiotic factor affecting the distribution and abundance of insects. Gut microbiota can contribute to host fitness, thereby increasing resistance to abiotic stress conditions. In this study, Sarcophaga peregrina is closely associated with human life in ecological habits and shows remarkable adaptability to daily and seasonal temperature fluctuations. To date, the role of gut microbiota in S. peregrina response to heat stress and its influence on the host phenotypic variability remain poorly studied. METHODS: We exposed S. peregrina to heat stress at 40 °C for 3 h every day throughout the developmental stages from newly hatched larva to adult, after which gut DNA was extracted from third-instar larvae, early pupal stage, late pupal stage, and newly emerged adults, respectively. Then, 16S rRNA microbial community analyses were performed. RESULTS: Firstly, we analyzed whether heat stress could have an impact on the life history traits of S. peregrina and showed that the growth rate of larvae was higher and the developmental time was significantly shorter after heat stress. We then proposed the role of the gut microbiota in the heat tolerance of S. peregrina, which indicated that the bacterial abundance and community structure changed significantly after heat tolerance. In particular, the relative abundance of Wohlfahrtiimonas and Ignatzschineria was higher in the third-instar larval larvae; the former increased and the latter decreased significantly after heat stress. To further explore the effect of disturbing the microbial community on thermotolerant phenotype, newly hatched larvae were fed with amikacin under heat stress, which indicated that the larval length and the whole developmental cycle was significantly shorter. CONCLUSION: This study indicated that Wohlfahrtiimonas and Ignatzschineria should play an important role in the post-feeding stage under heat stress, but further study is still needed. In general, heat tolerance can affect the gut microbial community structure, which in turn affects the fitness of the host.

Effects of Terbufos (Organophosphate) on Larval Behaviour of Two Forensically Important Diptera Species: Contributions for Entomotoxicology.

Neurotoxicant compounds interfere with the behaviour and biology of insects, significantly altering their locomotion patterns. However, little is known about the effect of organophosphates, neurotoxicants for agricultural, domestic and industrial use, on the larval movement of necrophagous flies, although being responsible for frequent cases of poisoning and accidental or intentional deaths. Thus, we aimed to study the influence of Terbufos (organophosphate) on the activity and mobility patterns of Lucilia eximia (Wiedemann 1819) (Calliphoridae) and Peckia (Peckia) chrysostoma (Wiedemann 1830) (Sarcophagidae) immatures collected from rat carcasses intoxicated with 5, 10 or 20 mg/kg of Terbufos, to evaluate (i) peristaltic movements and body contractions, and (ii) distance and shape of the trajectory travelled by the larva. Behavioural parameters were analysed in loco and through videos. We observed that the presence of Terbufos altered poisoned larvae's activity and body mobility in both taxon and dose-dependent manner. Lucilia eximia larvae were more active, with greater frequency of body movements and lateral contractions when intoxicated with high and intermediate doses of Terbufos. On the other hand, P. (P.) chrysostoma immatures were less active, with fewer body and lateral contractions when intoxicated with the high dose of the compound. This work experimentally demonstrates that the presence of Terbufos can alters the mobility and movement of intoxicated necrophagous Diptera, essential components of the cadaveric fauna.

Machine learning analysis of wing venation patterns accurately identifies Sarcophagidae, Calliphoridae and Muscidae fly species.

In medical, veterinary and forensic entomology, the ease and affordability of image data acquisition have resulted in whole-image analysis becoming an invaluable approach for species identification. Krawtchouk moment invariants are a classical mathematical transformation that can extract local features from an image, thus allowing subtle species-specific biological variations to be accentuated for subsequent analyses. We extracted Krawtchouk moment invariant features from binarised wing images of 759 male fly specimens from the Calliphoridae, Sarcophagidae and Muscidae families (13 species and a species variant). Subsequently, we trained the Generalized, Unbiased, Interaction Detection and Estimation random forests classifier using linear discriminants derived from these features and inferred the species identity of specimens from the test samples. Fivefold cross-validation results show a 98.56 ± 0.38% (standard error) mean identification accuracy at the family level and a 91.04 ± 1.33% mean identification accuracy at the species level. The mean F1-score of 0.89 ± 0.02 reflects good balance of precision and recall properties of the model. The present study consolidates findings from previous small pilot studies of the usefulness of wing venation patterns for inferring species identities. Thus, the stage is set for the development of a mature data analytic ecosystem for routine computer image-based identification of fly species that are of medical, veterinary and forensic importance.

Aphaereta pallipes (Hymenoptera: Braconidae) and Dirhinus anthracia (Hymenoptera: Chalcididae) associated with Peckia (Euboettcheria) collusor (Diptera: Sarcophagidae) in Brazil.

Flies of the Sarcophagidae family are widely distributed in the world, occupying different habitats. Some species have a high degree of synanthropy and are therefore often found in households in the urban environment. In Brazil, there is still little information related to the natural enemies of these insects in the urban environment, where population control is strictly chemical. Therefore, larvae and pupae of Peckia (Euboettcheria) collusor (Curran and Walley) (Diptera: Sarcophagidae) species were exposed in an urbanized location, and the presence and prevalence of parasitoids in the natural control of these immature stages was evaluated. We report for the first time the species Aphaereta pallipes (Say) (Hymenoptera: Braconidae) and Dirhinus anthracia Walker (Hymenoptera: Chalcididae), associated with P. (E.) collusor, highlighting the importance of these parasitoids in natural control in the urban environment, in addition to expanding the list of hosts for both parasitoid species and the distribution of this parasitoid-host interaction for Brazil and the Neotropical region.

Genome-wide analysis of long noncoding RNAs and their association in regulating the metamorphosis of the Sarcophaga peregrina (Diptera: Sarcophagidae).

BACKGROUND: The flesh fly, Sarcophaga peregrina (Diptera: Sarcophagidae), is an important hygiene pest, that causes myiasis in humans and other mammals, typically livestock, and as a vector for various parasitic agents, including bacteria, viruses, and parasites. The role of long non-coding RNAs (lncRNAs) in regulating gene expression during metamorphosis of the flesh fly has not been well established. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we performed genome-wide identification and characterization of lncRNAs from the early pupal stage (1-days pupae), mid-term pupal stage (5-days pupae), and late pupal stage (9-days pupae) of S. peregrina by RNA-seq, and a total of 6921 lncRNAs transcripts were identified. RT-qPCR and enrichment analyses revealed the differentially expressed lncRNAs (DE lncRNAs) that might be associated with insect metamorphosis development. Furthermore, functional analysis revealed that the DE lncRNA (SP_lnc5000) could potentially be involved in regulating the metamorphosis of S. peregrina. RNA interference of SP_lnc5000 caused reduced expression of metamorphosis-related genes in 20-hydroxyecdysone (20E) signaling (Br-c, Ftz-F1), cuticle tanning pathway (TH, DOPA), and chitin related pathway (Cht5). Injection of dsSP_lnc5000 in 3rd instar larvae of S. peregrina resulted in deformed pupae, stagnation of pupal-adult metamorphosis, and a decrease in development time of pupal, pupariation rates and eclosion rates. Hematoxylin-eosin staining (H&E), scanning electron microscope (SEM) observation and cuticle hydrocarbons (CHCs) analysis indicated that SP_lnc5000 had crucial roles in the metamorphosis developmental by modulating pupal cuticular development. CONCLUSIONS/SIGNIFICANCE: We established that the lncRNA SP_lnc5000 potentially regulates the metamorphosis of S. peregrina by putatively affecting the structure and composition of the pupal cuticle. This study enhances our understanding of lncRNAs as regulators of metamorphosis in S. peregrina, and provide valuable insights into the identification of potential targets for vector control and the development of effective strategies for controlling the spread of myiasis and parasitic diseases.

In vitro antibacterial activity and synergetic effect of crude extract of the Wohlfahrtia nuba (Diptera: Sarcophagidae) flesh fly larvae.

Multidrug-resistant pathogens have become ubiquitous, and effective treatment alternatives are urgently required. Maggot therapy is a promising agent that is being studied to overcome antibiotic-resistant pathogens. This study evaluated the antibacterial activity of the larvae extract of the Wohlfahrtia nuba (wiedmann) (Diptera: Sarcophagidae) flesh fly on the growth of five pathogenic bacterial species (methicillin-sensitive Staphylococcus aureus [ATCC 29213], methicillin-resistant Staphylococcus aureus [ATCC BAA-1680], Pseudomonas aeruginosa [ATCC 27853], Escherichia coli [ATCC 25922], and Salmonella typhi [ATCC 19430]) in vitro by using different techniques. Resazurin-based turbidimetric assay demonstrated that the W. nuba maggot exosecretion (ES) was potent against all the bacterial species tested, and according to the determined minimum inhibitory concentration (MIC) for each bacterium, gram-negative bacteria were more sensitive than gram-positive bacteria. Additionally, colony-forming unit assay showed that maggot ES was able to inhibit bacterial growth rate for all bacterial species tested, where the highest bacterial reduction was observed with methicillin-sensitive S. aureus (MSSA) followed by S. typhi. Moreover, maggot ES was shown to be concentration-dependent, where 100 µL of ES at 200 mg/mL was bactericidal towards methicillin-resistant S. aureus (MRSA) and P. aeruginosa compared with 100 µL at the MIC of the ES. Moreover, based on the result of agar disc diffusion assay, maggot extract was more efficient against P. aeruginosa and E. coli than the remaining reference strains tested. Furthermore, the combination between regular antibiotics with maggot ES at different concentrations indicated that ES acts synergistically with the tested antibiotics against the five bacterial models.

Identification of medically and forensically relevant flies using a decision treelearning method.

Blow flies, flesh flies, and house flies can provide excellent evidence for forensic entomologists and are also essential to the fields of public health, medicine, and animal health. In all questions, the correct identification of fly species is an important initial step. The usual methods based on morphology or even molecular approaches can reach their limits here, especially when dealing with larger numbers of specimens. Since machine learning already plays a major role in many areas of daily life, such as education, business, industry, science, and medicine, applications for the classification of insects have been reported. Here, we applied the decision tree method with wing morphometric data to construct a model for discriminating flies of three families [Calliphoridae, Sarcophagidae, Muscidae] and seven species [Chrysomya megacephala (Fabricius), Chrysomya rufifacies (Macquart), Chrysomya (Ceylonomyia) nigripes Aubertin, Lucilia cuprina (Wiedemann), Hemipyrellia ligurriens (Wiedemann), Musca domestica Linneaus, and Parasarcophaga (Liosarcophaga) dux Thomson]. One hundred percent overall accuracy was obtained at a family level, followed by 83.33% at a species level. The results of this study suggest that non-experts might utilize this identification tool. However, more species and also samples per specimens should be studied to create a model that can be applied to the different fly species in Thailand.

A Case of Nasopharyngeal Myiasis Caused by Sarcophaga sp

Myiasis is a parasitic infestation of tissues and body cavities of vertebrates by Diptera larvae. Nasal myiasis is mostly caused by the Sarcophagidae and Calliphoridae families. Many predisposing factors play a role in the appearance of nasal myiasis. In the treatment of the disease, the use of antiseptics together with mechanical cleaning is recommended. In this report, a case of nasopharyngeal myiasis in a 75-year-old patient hospitalized in the intensive care unit is presented. Nine larvae were detected in the patient's nose. Larvae were removed from the area and were cleaned with iodine solution for three days. As a result of macroscopic and microscopic examination, the larvae were identified as the third stage of Sarcophaga sp. in order to prevent myiasis infestations, personnel working in places where the patient is unconscious, such as intensive care units, should daily check the patient's body cavities such as nose, mouth and ear cavities, and perform their care and cleaning.

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.

On the life cycle of Brachymeria podagrica (Fabricius, 1787) (Hymenoptera: Chalcididae) - a parasitoid of sacrophagid, calliphorid, and muscid flies.

The lifecycle of Brachymeria podagrica, a parasitic wasp with a worldwide distribution, was studied under laboratory conditions using the flesh fly, Sarcophaga dux, as a host. Two hundred parasite-free 3rd instars of S. dux were exposed for 24 h to 20 female B. podagrica. In daily intervals, maggots and later pupae were examined for developmental stages of the parasitoid. The whole pre-imaginal development at a temperature of 26 °C lasted 21 to 26 days. Three morphologically different instars, followed by a prepupal and a pupal stage, were described using light and scanning electron microscopy. In a second experiment with 100 3rd stage Sarcophaga larvae and 10 parasitoids, a total of 70 wasps emerged 20 to 25 days after exposure. Two fly larvae did not pupate and dried out, while 28 pupae contained a dry or caseous content, dead wasp imagos, or their larval stages. No fly imagines emerged from exposed groups, while all 100 unexposed larvae pupated and adults eclosed between day 12 and day 14 after the start of the experiment, while the imagoes of the parasitoids appeared 8 to 12 days later.

Flesh Flies (Diptera: Sarcophagidae) of Forensic Importance Collected From Pig Carcasses in the Cerrado of Northeastern Brazil.

Sarcophagidae is a family of true flies frequently found colonizing carcasses and corpses as adults and/or larvae, therefore, they are of great forensic value in the medico-legal field. Thus, this study aims to inventory the sarcophagids species and describe their succession process in the decomposition of swine carcasses in the Cerrado (savanna-like vegetation), Brazilian Northeastern region. A total of 10,819 specimens were collected, belonging to 45 sarcophagid species. Of this total, one species was recently described (Helicobia neuzalmeidae Silva, Brasil, De-Souza & Carvalho-Filho), two [Blaesoxipha (Acanthodotheca) denieri (Blanchard) and Dexosarcophaga pusilla Lopes] are new records for the Cerrado biome, five [Argoravinia (Argoravinia) catiae Carvalho Filho & Esposito, B. (A.) denieri, Nephochaetopteryx orbitalis (Curran & Walley), Peckia (Sarcodexia) tridentata (Hall) and Tricharaea (Sarcophagula) ramirezi (Lopes)] are new records for the Northeast region and eight are new records for the state of Maranhão. The species A. (A.) catiae, Blaesoxipha (Gigantotheca) stallengi (Lahille), D. pusilla, H. neuzalmeidae, Lipoptilocnema misella Lopes, Oxysarcodexia bakeri (Aldrich) and Retrocitomyia retrocita (Hall) were collected for the first time in pig carcasses. Peckia (Sarcodexia) lambens (Wiedemann) and Oxysarcodexia thornax (Walker) were the most abundant species. In both collection seasons, five species were carcasses pioneers at an early stage. The majority of species (42 spp.) were collected during the dry season, primarily during the black putrefaction (39 spp.) and butyric fermentation (30 spp.) stages. The greatest number specimens (n = 6,484) were recorded during the rainy season, at the butyric fermentation (n = 2,517) and black putrefaction (n = 2,080).

Multimethod combination for age estimation of Sarcophaga peregrina (Diptera: Sarcophagidae) with implications for estimation of the postmortem interval.

Sarcophaga peregrina (Robineau-Desvoidy, 1830) (Diptera: Sarcophagidae) is a forensically important flesh fly with potential value for estimating the minimum postmortem interval (PMImin). Here, the developmental patterns of S. peregrina were investigated at 5 constant temperatures (15-35 °C). Morphological changes at different developmental stages of the pupa were observed at 4 constant temperatures (15-30 °C) by removing the puparium and staining the pupa with hematoxylin and eosin. Furthermore, differentially expressed genes (DEGs) were analyzed at 25 °C in the intrapuparial period to estimate the age of S. peregrina during the intrapuparial stage. S. peregrina completed development from larviposition to adult eclosion at 15 °C, 20 °C, 25 °C, and 30 °C; the developmental durations were 1090.3 ± 30.6 h, 566.6 ± 21.9 h, 404.6 ± 13.01 h, and 280.3 ± 4.5 h, respectively, while the development could not be completed at 35 °C. The intrapuparial period of S. peregrina was divided into 12 sub-stages on the basis of the overall external morphological changes; 6 sub-stages on the basis of individual morphological structures such as the compound eyes, antennae, thorax, legs, wings, and abdomen; and 10 sub-stages on the basis of internal morphological changes detected using histological analysis. The period of each sub-stage or structure that appeared was determined. Moreover, we found that 6 genes (NDUFS2, CPAMD8, NDUFV2, Hsp27, Hsp23, and TPP) with differential expression can be used for the precise age estimation of S. peregrina during the intrapuparial period. This study provided basic developmental data for the use of S. peregrina in PMImin estimation, and we successfully estimated PMImin in a real forensic case by using a multimethod combination.