Development and intrapuparial characterization of Peckia (Euboettcheria) collusor (Curran and Walley, 1934) (Diptera: Sarcophagidae) for application in forensic entomology.

The family Sarcophagidae is very diverse in Brazil. Due to their living habits, they are the subject of many medical, veterinary, sanitary, and entomological studies. However, Sarcophagidae species are still poorly studied in forensic entomology, although they are frequently reported in carcasses and even human corpses. Thus, this study aims to identify and compare the developmental stages and intrapuparial morphological characteristics of Peckia (Euboettcheria) collusor to serve as an auxiliary tool in forensic entomology. The pupae collected after zero hour at 27 °C and 32 °C were sacrificed every three hours until the first 24 h and then every six hours until the emergence of the first adults, using 30 pupae each time, totaling 1560 for 27 °C and 1290 for 32 °C. The intrapuparial development time of this fly species under laboratory-controlled conditions was 288 h at 27 °C and 228 h at 32 °C. The 2820 pupae were analyzed according to temperature and classified into eight possible stages. This contributed to the selection of 16 key morphological characteristics to identify the age of the pupae. The identified intrapupal morphological characteristics have great potential to help researchers, experts, technical assistants, and forensic entomologists estimate the minimum post-mortem interval (minPMI) of cadavers.

The gut bacterial composition across life stages of Sarcophaga peregrina (Diptera: Sarcophagidae) and the effects of amikacin on their development.

Insects and microorganisms, ubiquitous organisms in the natural world, have developed intricate relationships throughout their evolutionary histories. However, most studies have concentrated on specific time points or life stages, but some limited studies have investigated the dynamics of microbial diversity within insects across life stages. Here, 16S rDNA sequencing technology was used to investigate the gut bacterial community across the life stages of Sarcophaga peregrina (Robineau-Desvoidy) (Diptera: Sarcophagidae). The results revealed that the gut bacterial diversity of S. peregrina varied with life stage and showed similarity in the nearby life stages. Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes were the dominant phyla in S. peregrina. Genera such as Providencia, Ignatzschineria, and Myroides are implicated in potentially pivotal roles during the developmental processes of this flesh fly. Furthermore, the effects of amikacin on the growth and development of S. peregrina were not statistically significant. However, we did observe significant changes at the protein level, which suggests a close association between protein-level alterations and growth and development. Additionally, we speculate that S. peregrina regulates its nutritional status during nonfeeding stages to meet the demands of eclosion. This study represents the first comprehensive examination of the intestinal bacterial composition across various life stages of S. peregrina. Our findings deepen our understanding of the gut microbiota in this flesh fly and lay the groundwork for further exploration into the intricate interactions between microorganisms and insects.

Intrapuparial development of Sarcophaga (Liopygia) ruficornis (Diptera: Sarcophagidae), a species of medical-veterinary and forensic importance, under laboratory conditions.

Sarcophaga (=Parasarcophaga) (Liopygia) ruficornis (Fabricius, 1794) is a species of medical-veterinary and forensic importance, as its immatures cause myiasis in humans and animals and colonize carcasses and cadavers. Therefore, investigations into the biology and morphology of this species, with a particular focus on pupae that constitute ≥50% of the developmental period for the immatures, are pertinent. Although there are biological and morphological studies of pupae, the intrapuparial development at different temperatures has not yet been analysed. Therefore, the present study aimed to describe how temperature (22, 27 and 32 ± 1°C) affects the development and morphology of S. (L.) ruficornis pupae at 60 ± 10% relative humidity and a 12-h photoperiod. Ten pupae were collected, euthanized and fixed every 4 h from pupariation until 24 h and every 8 h until the emergence of the first adult. Emergence occurred at 440, 272 and 232 h at 22, 27 and 32°C, with 590, 380 and 330 pupae, respectively. The highest mortality occurred at 32°C. Eight periods were defined, and sex was determined in pharate adult stage; in addition, 40 key morphological characteristics to estimate pupal age were presented. These findings can serve as a basis for studies on the biology and morphology of the pupa of S. (L.) ruficornis, particularly in bionomics, control and forensics, helping researchers and experts.

Sarcophaga (=Parasarcophaga) (Liopygia) ruficornis (Fabricius, 1794) é uma espécie de importância médico­veterinária e forense, pois seus imaturos causam miíase em humanos e animais e colonizam carcaças e cadáveres. Portanto, são pertinentes as investigações sobre a biologia e morfologia dessa espécie, com foco especial nas pupas que constituem ≥50% do período de desenvolvimento dos imaturos. Embora existam estudos biológicos e morfológicos das pupas, o desenvolvimento intrapuparial em diferentes temperaturas ainda não foi analisado. Logo, o presente estudo teve como objetivo descrever como a temperatura (22, 27 e 32 ± 1°C) afeta o desenvolvimento e a morfologia das pupas de S. (L.) ruficornis a 60 ± 10% de umidade relativa e fotoperíodo de 12 horas. Dez pupas foram coletadas, eutanasiadas e fixadas a cada quatro horas desde a pupariação até 24 horas e a cada oito horas até a emergência do primeiro adulto. A emergência ocorreu em 440, 272 e 232 horas a 22, 27 e 32°C, com 590, 380 e 330 pupas, respectivamente. A maior mortalidade ocorreu a 32°C. Foram definidos oito períodos e o sexo foi determinado na fase de adulto farato; além disso, foram apresentadas 40 características morfológicas importantes para estimar a idade das pupas. Essas descobertas podem servir de base para estudos sobre a biologia e a morfologia da pupa de S. (L.) ruficornis, especialmente em bionomia, controle e ciência forense, ajudando pesquisadores e especialistas.

Myiasis in humans: Case reports in Northeastern Brazil including multispecies co-infestation by Sarcophagidae.

Myiasis is a parasitosis characterized by an infestation of living vertebrates (humans and other animals) by Diptera larvae, whose occurrence and etiological identification are still neglected by health professionals. Here we analyzed the human myiasis cases registered from 2010 to 2018 in health care units in the municipality of Natal, Northeast region of Brazil. Specifically, we aimed to: I) analyze the medical records of cases documented from 2010 to 2017; II) list the patients predisposing factors; and III) monitor the recent cases diagnosed in health units between August 2017 and March 2018 and report the taxonomic identity of the infesting species. Our data revealed that myiases mainly affect the elderly and individuals with predisposing medical conditions (e.g. senility, filariasis). Regarding the new cases, larvae of Calliphoridae (C. hominivorax (Coquerel, 1858)) and Sarcophagidae [Sarcophaga (Liopygia) ruficornis (Fabricius, 1794), Peckia (Sarcodexia) lambens (Wiedemann, 1830), and Helicobia morionella (Aldrich, 1930)] were identified. Furthermore, this was the first report of a co-infestation with three Sarcophagidae species and also the presence of H. morionella in a human wound. These results reinforce that myiasis is an underreported parasitosis, which may underestimate the ability of previously unreported fly species to feed on living human tissues in the Neotropical Region.

Gingival Myiasis on Oral Squamous Cell Carcinoma in Turkey: A Case Report

Myiasis is a disease caused by tissue invasion of diptera larvae and eggs. Oral myiasis is mostly related to old age, poor oral hygiene, suppurative lesions, anatomical disorders and cancer cases. Oral squamous cell carcinoma (OSCC) is an important risk factor for myiasis. This report presents the case of an 82-year-old woman who presented with gingival myiasis developing on the background of OSSC. The patient was diagnosed with OSSC in the hospital. Myiasis larvae were identified and sent to the National Parasitology Reference Laboratory for identification. Thus, development of myiasis on OSCC background was shown in Turkey for the first time. Myiasis larvae have been identified as the 3rd phase of the larvae Sarcophaga sp. development (Diptera: Sarcophagidae). As a result, myiasis cases are sporadic in Turkey, and it can be avoided by controlling fly population and by paying attention to hygiene. Controlling myiasis is an important public health problem and should be considered in a single health concept, as it causes health problems in both humans and animals. The findings of this case will draw attention to the importance of dealing with myiasis factors, which is a public health problem.

Effect of Terbufos (Organophosphate) on the Cadaveric Colonization Process: Implications for Postmortem Interval Calculation.

The determination of necrophagous fly specie's development time is considered an accurate method for estimating postmortem interval (PMI). However, pesticides and other chemicals can alter the flies' life cycle, inducing errors in PMI estimation. Thus, this work aimed to evaluate the effect of different doses of Terbufos (Organophosphates) on the temporal dispersion pattern and development of immature dipterans associated with decaying rat carcasses. For this, 150 g female Wistar rats received, via gavage, 200 µl of Terbufos (5 or 10 mg/kg) or distilled water (control) and, after 30 min of administration, the animals were euthanized and distributed in suspended traps to decompose under environmental conditions. The dispersing immatures were collected daily, and their development time was monitored until the emergence of adult flies. After data analysis, it was observed that Terbufos altered 1) the temporal pattern of larval dispersion; 2) the composition and structure of the colonizing assemblage (emerged adults); 3) species' development time, accelerating or delaying their cycle, depending on the dose used; and 4) the califorids and sarcophagids emergence rate, increasing the mortality of pupae from intoxicated carcasses. Thus, this work demonstrates experimentally that Terbufos directly influences the development of flies with forensic potential and discusses the implications for PMI estimation, which can assist in future investigative processes with suspected poisoning by this organophosphate.

Effect of constant and fluctuating temperature on the intrapuparial development of Sarcophaga argyrostoma (Robineau-Desvoidy, 1830; Diptera: Sarcophagidae).

This study aimes to investigate the effect of constant and fluctuating temperatures on starting times of developmental periods of insect species. For this purpose, Sarcophaga (Liopygia) argyrostoma (Robineau-Desvoidy, 1830), a forensically important species has been chosen. Whether the constant (25°C) and fluctuating (25.2°C averaged over natural conditions) temperatures would make a difference on the timing of intrapuparial development of this species was investigated. First larval instars were placed in rearing cages until they reached post-feeding third larval instar, which is considered the beginning of intrapuparial development. Relevant morphological changes were observed by hourly dissections. At fluctuating temperature treatment, the post-feeding third instar larvae were transferred to containers under natural, open-air conditions. Minimum and maximum starting times of 23 intrapuparial development periods were determined for both treatments. These times, medians, and standard deviations were compared by Mann-Whitney U tests. Accumulated degree hour values were also calculated for these periods. Regardless of temperature treatment, intrapuparial development periods were similar and adult emergence was observed at 289.5 ( ± 1.52) h under 25°C constant temperature and 319 ( ± 1.41) h under fluctuating temperature conditions. No statistical difference was found between the intrapuparial development of S. argyrostoma reared under constant and fluctuating temperatures, except "pupal-adult apolysis" and "checkered pattern on the abdomen becoming distinct" periods. The use of the results obtained when determining the effect of constant and fluctuating temperatures on the starting times of developmental periods of this species in forensic entomology, is also important in transferring developmental biology data into the field of applied sciences.

Development of Sarcophaga dux (diptera: Sarcophagidae) at constant temperatures and differential gene expression for age estimation of the pupae.

Sarcophaga dux (Diptera: Sarcophagidae) is a necrophagous flesh fly species with potential forensic value for estimating minimum postmortem interval (PMImin). The basic developmental data and precise age estimates of the pupae are significant for PMImin estimation in forensic investigations. In the present study, we investigated the development data of that species at seven constant temperatures varying from 16 °C to 34 °C, including body length changes of the larve, developmental duration and accumulated degree hours of the preadults. Several reference genes for relative quantification of the differentially expressed genes (DEGs) were firstly selected and evaluated in the pupae of different ages under different temperatures. The DEGs of the insects during the pupal period at different constant temperatures (34, 25 and 16 °C) were further analyzed for more precise age estimation. The results showed that the developmental durations of the preadults at 16, 19, 22, 25, 28, 31 and 34 °C were 1478.6 ± 18.3 h, 726.1 ± 15.8 h, 538.5 ± 0.9 h, 394.1 ± 9.5 h, 375.6 ± 10.8 h, 284.1 ± 7.3 h, and 252.5 ± 6.1 h, respectively. The developmental threshold temperature the flies was 12.27 ± 0.35 °C, and the thermal summation constant was 5341.71 ± 249.29° hours. The most reliable reference genes during the pupal period at different temperatures were found: GST1 and 18S rRNA for the 34 °C group, GST1 and RPL49 for 25 °C, and 18S rRNA and 28S rRNA for 16 °C. The four differential expression genes (Hsp60, A-alpha, ARP, and RPL8) have the potential to be used for more precise age estimation of pupal S. dux. This work provides important basic developmental data and a more precise age estimation method for pupal S. dux, and improves the value of this species for PMImin estimation in forensic investigations.

The colonisation of remains by the sarcophagid fly Sarcophaga argyrostoma (Robineau-Desvoidy) (Diptera: Sarcophagidae).

This article discusses the ability of the sarcophagid fly Sarcophaga argyrostoma (Robineau-Desvoidy) to exploit various food sources. These include living, freshly dead, and highly decayed remains. It also considers the factors affecting larval thermogenesis and compares S. argyrostoma with Lucilia sericata (Meigen). In addition, the ability of S. argyrostoma to colonise buried baits, those out of reach, and those protected by bandaging is considered. Both fresh and decayed liver stimulated ovoviviposition by S. argyrostoma and oviposition by L. sericata. Even liver that had decayed for 840 days supported larval development by both species to adulthood. Thermogenesis during larval development was dependent upon larval density. There was no difference in thermogenesis characteristics between S. argyrostoma and L. sericata or between diets of fresh liver and fresh minced pork. Thermogenesis was less pronounced on 840-day-old liver than on fresh liver but was still dependent upon larval density and there was no species effect. When starved, third instar S. argyrostoma larvae did not exhibit cannibalism or attack third instar L. sericata larvae. Sarcophaga argyrostoma adults could detect fresh liver baits buried at 20 cm in loose soil and ovoviviposited on the soil surface. The larvae then rapidly crawled through the soil to reach the baits and completed their development below ground. Sarcophaga argyrostoma also colonised baits that were at least 5 cm out of reach by dropping larvae onto them. They could infest baits through gauze bandages but not wound dressings. The adults would not ovoviviposit on living invertebrates but dead snails and dead L. sericata third instar larvae were infested and supported larval growth to adulthood. The adults were willing to ovoviviposit on human faeces and it supported larval growth to adulthood. Human urine stimulated very limited ovoviviposition and the larvae all died shortly afterwards and without developing. They did not ovoviviposit on banana or melon.

Developmental Plasticity of the Flesh Fly Blaesoxipha plinthopyga (Diptera: Sarcophagidae) on Different Substrates.

Forensic entomologists rely on laboratory growth data to estimate the time of colonization on human remains thus extrapolating a minimum postmortem interval (PMI) if assumptions are satisfied. The flesh fly Blaesoxipha plinthopyga (Wiedemann) is one species that occurs in casework in Idaho, Texas, and central California. Because of the few laboratory studies on the development of this fly, the following study was conducted to determine if different substrates impact immature development of the species. In this study, flies were reared on different substrates that are likely to be encountered at indoor and outdoor scenes (Wet Sand, Dry Sand, Clothes [Polyester fibers], and Carpet [Polypropylene fibers]) to determine the influence of substrate on larval, intrapuparial, and total immature development times at 25°C, 50% RH, and 14:10 (L:D) h cycle. The results revealed that substrate significantly affected minimum immature development times without affecting the sexes differently; though a female bias in sex ratio was observed consistently. Average minimum larval developmental times were 160-179 h with a significantly faster development in Carpet than in Clothes. Similarly, average minimum intrapuparial developmental times were 331-352 h; fastest on Carpet and the slowest in Dry Sand. For this species, it may be important to consider the substrates encountered at a death scene as they may affect the development of B. plinthopyga (Wiedemann) in casework by up to 29 h at 25°C and 50% humidity. These effects will also be important to consider when planning future development studies with the species.

Differential Gene Expression for Age Estimation of Forensically Important Sarcophaga peregrina (Diptera: Sarcophagidae) Intrapuparial.

Sarcophaga peregrina is an important flesh fly species for estimating the minimum postmortem interval (PMImin) in forensic entomology. The accurate determination of the developmental age is a crucial task for using necrophagous sarcophagids to estimate PMImin. During larval development, the age determination is straight forward by the morphological changes and variation of length, weight, and width; however, the age estimation of sarcophagid intrapuparial is more difficult due to anatomical and morphological changes not being visible. The analysis of differentially expressed genes (DEGs) during sarcophagid metamorphosis is a potential method for age estimation of intrapuparial. In the present study, real-time quantitative polymerase chain reaction (RT-qPCR) was used to analyze the differential gene expression level of S. peregrina intrapuparial in different constant temperatures (35°C, 25°C, and 15°C). In addition, the appropriate reference genes of S. peregrina were selected in the intrapuparial and at different temperatures to obtain reliable and valid gene expression profiles. The results indicated that two candidate genes (18S rRNA and 28S rRNA) were the most reliable reference genes, and four DEGs (Hsp90, A-alpha, AFP, AFBP) have the potential to be used to more accuracy estimate the age of S. peregrina intrapuparial.

A Study of the pupal developments of Sarcophaga argyrostoma (Robineau-Desvoidy, 1830).

Forensic entomology is a discipline in which insect evidence is used to determine the minimum postmortem interval (PMImin). The insect evidence may also provide valuable information about the movement of the body, cause of the death, drug use, or poisoning. The pupal period constitutes approximately 50% of immature development. Therefore, it provides a more accurate estimation of the PMImin via a detailed examination. In this study, the pupal development of Sarcophaga (Liopygia) argyrostoma (Robineau-Desvoidy, 1830), which larviposits on dead bodies, was examined in Turkey. The pupal development periods of S. argyrostoma and the range of the beginning hours (maximum and minimum) of these periods were determined at a constant temperature of 20, 25, and 30 °C. When all of the temperatures were considered, it was determined that 23 pupal development stages were similar; however, the development times shortened as the temperature increased. Adult emergence was observed at 459 (±1.41), 289.5 (±1.52), and 227 (±1.89) h at 20, 25, and 30 °C, respectively. With respect to the pupal period used to determine the PMImin, the development of S. argyrostoma at different temperatures was described in this study for the first time.

Geographical Distribution, Biodiversity, and Species Richness of Medically Important Necrophagous Flies in Central Iran.

There have been some investigations carried out about necrophagous flies and their biodiversity indices in Iran. In the present study, the species composition of flies, their richness, and biodiversity were investigated in central Iran. In a cross-sectional study, specimens were collected monthly in three different climate areas of Qom Province. Species diversity, species richness, and species evenness were calculated to estimate the species biodiversity of flies. In total, 7,291 flies were collected and identified using valid identification keys that classified flies into four families and 15 different species. The largest collection of flies was obtained in the mountainous areas comprising 36.2% of specimens. Moreover, five forensically important species of fly were collected in this region with the potential to cause human and animal myiasis: Calliphora vicina, Lucilia sericata, Musca domestica, Wohlfahrtia nuba, and Chrysomya albiceps. There were slight differences in species richness and diversity indices in the areas. In mountainous areas, the Shannon-Wiener index and richness were maximum (H' = 1.87, S = 13). The evenness based on the Shannon evenness index in the mountainous areas was 0.728 and 0.719 in desert areas, whereas it was the highest (0.764) in the semidesert areas. Moreover, high values of the Jaccard similarity index were observed in captured necrophagous fly species in all the areas. The necrophagous fly fauna and species diversity indices were high because of an increase in the evenness. In general, some well-known forensically important species of flies were collected that proved the potential of myiasis occurrence in this area.

A formulation of neem and hypericum oily extract for the treatment of the wound myiasis by Wohlfahrtia magnifica in domestic animals.

Untreated, traumatic, or wound myiases can cause severe consequences to animal health and welfare as well as economic losses to livestock productions. For healing myiasis-caused wounds, disinfectant such as creolin is wrongly but currently used in association with insecticides. Though effective, creolin is highly toxic to the patients, is inadequate with respect to the repellent effect, and may delay the healing of treated wounds. In this uncontrolled study, the efficacy of the patented plant-derived formulation 1 Primary Wound Dressing© (1-PWD), composed of neem oil (Azadirachta indica A. Juss.) and the oily extract of Hypericum perforatum (L.) flowers, has been investigated. Forty-four domestic animals of different species suffering from wound myiasis lasting for up to 25 days, at different parts of the body, were enrolled in the study. No systemic or local antibiotic or disinfectants' treatment was administered. Larvae recovered on open wounds and adults reared from mature larvae were identified as Wohlfahrtia magnifica. All the treated wounds healed in a range of 10 to 32 days without further infestation. None of the recruited animals presented bacterial complications. Data herein presented indicate that the tested natural plant-derived formulation is able to manage the infestation caused by W. magnifica larvae and the healing process of traumatic infested wounds in several domestic animal species, without any side effect on the living tissue and without the need to use local or systemic chemical or other products.

Molecular identification and detection of Wohlfahrtia magnifica in ovine vulvar myiasis in Gansu, China.

Traumatic myiasis causes substantial economic losses to farmers worldwide. In the present study, six flocks of sheep (2261 sheep) were investigated in Gansu, China, and 207 of 552 larvae were genetically characterized based on three genes, including cyt b, EF-1α, and white gene, by polymerase chain reaction and sequence analysis. A survey of sheep in China revealed that the prevalence of vulvar myiasis of six sheep flocks was 5.00% (11/220, Flock1), 4.85% (10/206, Flock2), 4.50% (9/200, Flock3), 5.00% (15/300, Flock4), 4.68% (15/320, Flock5), 0% (0/1015, Flock6), respectively. The sequence and phylogenetic analysis showed that only Wohlfahrtia magnifica was detected in the field samples. This is the first report of ovine vulvar myiasis caused by W. magnifica in Gansu, China. Some prophylactic measures are strongly recommended to reduce the risk of sheep acquiring traumatic myiasis in Gansu, China.

Forensically Important Boettcherisca peregrina (Diptera: Sarcophagidae) in China: Development Pattern and Significance for Estimating Postmortem Interval.

Sarcosaprophagous flesh flies are insect species frequently found on corpses, and their developmental patterns can be used as reliable indicators for estimating minimum postmortem interval (PMImin). In this study, the Boettcherisca peregrina Robineau-Desvoidy was reared at seven constant temperatures ranging from 16 to 34 °C to investigate the indexes of development duration and larval body length. Using these results, we generated three development models, including isomorphen diagram, isomegalen diagram, and thermal summation model. Regression analysis was conducted to obtain simulation equations of the variation in larval body length with time after larviposition, and variation in time after larviposition with body length. The developmental durations of B. peregrina from larviposition to adult eclosion under 16, 19, 22, 25, 28, 31 and 34 °C were 1,064.7 ± 34.8, 756.0 ± 19.0, 559.6 ± 5.5, 414.3 ± 3.9, 315.0 ± 2.0, 278.0 ± 4.0, and 258.0 ± 3.5 h, respectively. The developmental threshold temperature of B. peregrina was 10.87 ± 0.49 °C, and the thermal summation constant was 5,809.7 ± 291.4 degree days. The results of this study provide basic data for the use of B. peregrina for estimating PMImin.

Molecular phylogeny of Miltogramminae (Diptera: Sarcophagidae): Implications for classification, systematics and evolution of larval feeding strategies.

Miltogramminae is one of the phylogenetically most poorly studied taxa of the species-rich family Sarcophagidae (Diptera). Most species are kleptoparasites in nests of solitary aculeate wasps and bees, although parasitoids and saprophagous species are also known, and the ancestral miltogrammine life habit remains unsettled. Here, we present for the first time a comprehensive phylogenetic tree consisting of 58 representatives of Miltogramminae, reconstructed using sequence data from three mitochondrial (COI, cytB, ND4) and one nuclear (Ef-1α) genes. Our phylogenetic hypothesis suggests that: (1) Miltogramminae are sister to Paramacronychiinae, (2) Miltogramminae can be divided into the "lower miltogrammines" containing two clades of mainly saprophages and a clade of "higher miltogrammines" with mainly kleptoparasitic species, (3) only three genera turn out to be non-monophyletic: Miltogramma, Senotainia and Pterella and (4) the genus Sarcotachina, which traditionally has been considered as belonging to the Paramacronychiinae, is placed in one of the clades of "lower miltogrammines". Ancestral state reconstruction of larval feeding strategy and five larval characters reveals that the ancestor of Miltogramminae was likely a saprophage retaining plesiomorphic oral ridges and a cephaloskeleton with sclerotized dorsal bridge. Synapomorphies like large pseudocephalic sensory organs and well-developed cuticular sculpture suggest that the ancestral first instar larva actively searched for a buried food supply.

Insecticidal activity of Stemona collinsiae root extract against Parasarcophaga ruficornis (Diptera: Sarcophagidae).

In Thai indigenous knowledge, Stemona plant has traditionally been used as insecticide on plantations. Recently, S. collinsiae extract was showed to be an anti-feedant and growth inhibitor and to exert strong insecticidal activity. Here, the insecticidal activity of S. collinsiae root extract against Parasarcophaga ruficornis is studied. The larvicidal and pupicidal activities of the ethanolic root extract of S. collinsiae were tested using contact toxicity tests, and adulticidal activity was tested using the topical contact toxicity and sugar bait methods The ethanolic extract at concentration ranging of 0.3-320mg/larva for the direct contact toxicity test and from 0.3 to 3.2mg/cm2/larva for the secondary contact toxicity test showed 3.0-51.0 and 1.0-94.0% corrected mortality, respectively. Against third-instar larvae, the LD50 concentrations of the ethanolic extract were 31.7±0.0mg/larva and 1.4±0.0mg/cm2/larva for direct and secondary contact toxicity tests, respectively. Pupae were not eliminated at all concentrations of the ethanolic extract. Against adult flies, which were killed via oral administration, the LD50 concentration of the ethanolic extract was 0.145±0.070g extract/g glucose. Thus, the ethanolic extract of S. collinsiae was capable of eliminating P. ruficornis in larval and adult stages via topical and ingestion administration, respectively.

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.

THE EFFECT OF THE TRAMADOL ACCUMULATED IN RAT LIVER ON THE DEVELOPMENT OF THE IMMATURE STAGES OF THE FLESH FLY SARCOPHAGA ARGYROSTOMA (ROBINEAU-DESVOIDY, 1830) (DIPTERA: SARCOPHAGIDAE).

The present study investigated the effects of the accumulation of tramadol on the development rates of the larvae and pupae. Larvae of the flesh fly Sarcophaga argyrostoma (Robineau-Desvoidy, 1830) were reared on three groups of Wister albino rat livers. One group was administered the recommended or normal dose (Dl) and the second group was injected with higher over or double dose (D2) of tramadol. Tramadol was administered by stomach tube once a day, for 3 months. The third group was injected with normal saline solution as the control. Using GC-Ms analysis, Rat livers contained 0.72 and 1.62mg/g of tramadol, in case of Dl and D2, respectively. Larvae of S. argyrostoma fed on DI and D2 rat livers contained (0.11 & 0.1 8mg/g), respectively. The corresponding tramadol concentrations persisted in the produced pupae of S. argyrostoma were (0.07 & 0.09mg/g), respectively. The accumulation of tramadol in tissues of S. argyrostoma larvae reduced the durations of larval stadia from (7.0408 1.0198 days) in case of the control to (6.6383±0.4857 days) when fed on DI liver and (6.3438±0.4826 days) when fed on D2 livers. The pupal durations was altered to (14.3750±0.4919 days) for D2 and (14.9574±0.6580 days) for DI, compared to (13.9167±1.0071 days), in case of the control. The average weight of the 3rd day larva of S. argyrostoma decreased from 18.1807±1.4949mg, in case of the control to 14.7279±1.3366mg and 14.9560±3.8210mg, when fed on Dl and D2 tramadol treated livers. In contrast, corresponding weight produced pupae increased from 14.1750±0.4667mg (control). to 15.2449±0.302mg & 15.9062±0.2888mg, in case of D1 and D2, respectively.