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Transgenic insect strains with tetracycline repressible (Tet-Off) female-lethal genes provide significant advantages over traditional sterile insect techniques for insect population control, such as reduced diet and labor costs and more efficient population suppression. Tet-Off systems are suppressed by tetracycline-class antibiotics, most commonly tetracycline (Tc) or doxycycline (Dox), allowing for equal sex ratio colonies of transgenic insects when reared with Tc or Dox and male-only generations in their absence. Dox is a more stable molecule and has increased uptake than Tc, which could be advantageous in some insect mass-rearing systems. Here, we evaluated the suitability of Dox for rearing Tet-Off female-lethal strains of Australian sheep blowfly, Lucilia cuprina (Wiedemann, 1830) (Diptera: Calliphoridae), and New World screwworm, Cochliomyia hominivorax (Coquerel, 1858) (Diptera: Calliphoridae), and the effects of dosage on strain performance. For both species, colonies were able to be maintained with mixed-sex ratios at much lower dosages of Dox than Tc. Biological yields of C. hominivorax on either antibiotic were not significantly different. Reduction of Dox dosages in C. hominivorax diet did not affect biological performance, though rearing with 10 or 25 μg/mL was more productive than 50 μg/mL. Additionally, C. hominivorax mating performance and longevity were equal on all Dox dosages. Overall, Dox was a suitable antibiotic for mass-rearing Tet-Off female-lethal L. cuprina and C. hominivorax and was functional at much lower dosages than Tc., (Published by Oxford University Press on behalf of Entomological Society of America 2024.)

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., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America.)

Aging blow fly eggs can be critical to a forensic investigation, but there are currently no forensically useful timelines describing internal anatomical changes in embryological development. This is partly due to the lack of an economical, rapid and technically simple histological technique to allow mass production of slides for research and casework. We present a histological method that uses a slightly modified standard laboratory processing run with 1 h fixation in 10% formalin, 2 h softening in Molliflex and Haemotoxylin and Eosin (H&E) staining. We also present a summary of the internal anatomical changes that can be visualized using our technique in the developing eggs of Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae). We examined eggs from at least three different females grown at 15 °C and sampled at 6 h intervals, and eggs grown at 20 °C and sampled at 3 h intervals. Blind aging trials demonstrated that it is possible to accurately age material grown at 20 °C to within 6 h (but attempts to further narrow this interval resulted in errors in one-third of cases). It was also possible to see sufficient anatomical detail to age eggs preserved for forensic casework 5, 8, 9, 10 and 11 years previously. Additionally, we determined that section quality was improved by 5 s fixation in hot water prior to preservation in ethanol. However, hot water fixation for longer than this increased the level of section artefact.

Common DNA-based species determination methods fail to distinguish some blow flies in the forensically and medically important genus Lucilia Robineau-Desvoidy. This is a practical problem, and it has also been interpreted as casting doubt on the validity of some morphologically defined species. An example is Lucilia illustris and L. caesar, which co-occur in Europe whilst only L. illustris has been collected in North America. Reports that these species shared both mitochondrial and nuclear gene sequences, along with claims that diagnostic morphological characters are difficult to interpret, were used to question their separate species status. We report here that amplified fragment length polymorphism profiles strongly support the validity of both species based on both assignment and phylogenetic analysis, and that traditional identification criteria based on male and female genital morphology are more reliable than has been claimed.

This research aims to describe the process of ecological succession by associating the decomposition stages of pig carcasses with flies from the Calliphoridae family (Diptera). For this, 6 pig carcasses were exposed in Maranhão's Cerrado, utilizing metal cages with sawdust trays to catch immature specimens and "suspended traps" to capture adults. Adults of the Hemilucilia benoisti Séguy, and Hemilucilia townsendi Shannon species were only associated with the swelling stage. Chrysomya megacephala (Fabricius) was associated with 2 stages (black putrefaction and fermentation). The species Chloroprocta idioidea (Robineau-Desvoidy), Chrysomya albiceps (Wiedemann), Cochliomyia macellaria (Fabricius), and Lucilia eximia (Wiedemann) were associated with 4 of the 5 stages of decomposition, the latter being the only 1 associated with the initial stage. The larvae abandoned the carcasses to pupate from the second stage of decomposition, with L. eximia being the only 1 leaving the carcasses in the swelling stage, and C. albiceps the only 1 associated with both the fermentation and dry stages. Our findings indicate that calliphorid species can help forensic investigators estimine the post-mortem interval of cadavers in situations similar to those detailed in this study. Since there was a link between adult and immature species and certain stages, they can be used as indicators in future forensic investigations., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

We evaluated the effects of seasonality on the richness and abundance of dipterans of the families Calliphoridae and Mesembrinellidae associated with the decomposition of a clothed Large White swine Sus scrofa domesticus(Artiodactyla: Suidae) carcass. Experiments were carried out in less rainy, rainy, and intermediate periods between 2010 and 2011 at Reserva Florestal Ducke, Manaus, Amazonas. Two pig carcasses, each weighing approximately 40 kg, were used in each period. A total of 63,872 individuals of 18 species of Calliphoridae and Mesembrinellidae were collected. The abundance and richness of these dipteran families were influenced by the interaction between period and decomposition stage. The compositions of the Calliphoridae and Mesembrinellidae assemblages differed among periods, with the fauna of the less rainy period being less similar to those of the intermediate and rainy periods than they were to each other. Three species were selected as indicators for the less rainy period, namely Paralucilia pseudolyrcea (Mello, 1969) (Diptera, Calliphoridae), Paralucilia nigrofacialis (Mello, 1969) (Diptera, Calliphoridae), and Eumesembrinella randa (Walker, 1849) (Diptera,Mesembrinellidae) while Chloroprocta idioidea (Robineau-Desvoidy, 1830) (Dipetra, Calliphoridae) was selected as an indicator species for the rainy period; no taxon was selected as an indicator of the intermediate period. Among decomposition stages, only fermentation and black putrefaction had indicator taxa, with Hemilucilia souzalopesi Mello, 1972 (Diptera, Calliphoridae and Chysomya putoria(Wiedemann, 1830) (Diptera, Calliphoridae), respectively. Clothes did not prevent the laying of eggs and became a kind of protection for immature stages. The clothed model presented a delay in decomposition compared to other studies developed in the Amazon region., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Blow flies are of medical, sanitary, veterinary, and forensic importance. Their accurate taxonomic identification is essential for their use in applied research. However, neotropical fauna has not been completely studied or described, and taxa identification without the required training is a difficult task. Additionally, the current morphological keys are not fitting to all extant taxa. Molecular-based approaches are widely used to overcome these issues, including the standard 5' COI barcode fragment (~650 base pairs [bp]) for identification at the species level. Here, a shorter sequence of 5' COI fragment (~342 bp) was assessed for the identification of 28 blow fly species inhabiting the northwest of South America. One tree-based (the generalized mixed Yule-coalescent-GMYC) and 3 distance-based approaches (automatic barcode gap discover - ABGD, the best close match - BCM, and the nearest neighbor - NN) analyses were performed. Noticeably, the amplification and sequencing of samples that had been preserved for up to 57 years were successful. The tree topology assigned 113 sequences to a specific taxon (70% effectiveness), while the distance approach assigned to 95 (59% effectiveness). The short fragment allowed the molecular identification of 19 species (60% of neotropical species except for the Lucilia species and Hemilucilia semidiaphana). According to these findings, the taxonomic and faunistic considerations of the blow fly fauna were provided. Overall, the short fragment approach constitutes an optimal species confirmation tool for the most common blow flies in northwestern South America., (© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

New World screwworm flies, Cochliomyia hominivorax (Coquerel), are obligate parasites of warm-blooded animals. They were eradicated from North and Central America during the mid-20th to early-21st centuries using the sterile insect technique (SIT), a method presently employed to maintain a permanent barrier between Central and South America. Lures are an important component of the screwworm eradication program, where they are used for surveillance, sample collection, and strain evaluation in the field. The first chemical lure, later named swormlure, was developed based on the attractiveness of C. hominivorax to volatile organic compounds (VOCs) produced from decomposing animal tissues. The formulation has changed little over the years and presently contains 10 chemicals, one of which is dimethyl disulfide (DMDS). Restrictions on the transport of DMDS have recently impeded its use in swormlure-4 (SL-4). However, dimethyl trisulfide (DMTS) is not as highly restricted and can be shipped via air transportation. Both chemicals are produced by microbial decomposition of animal tissues. Here, we conducted field trials using three releases of sterile C. hominivorax, each comprising approximately 93,000 flies, to test the efficacy of SL-4, containing DMDS, to swormlure-5 (SL-5) containing DMTS. Traps baited with SL-4 and SL-5 captured 575 (mean = 191.7, SD 17.9) and 665 (mean = 221.7, SD 33.2) C. hominivorax, respectively (df = 19, F = 1.294, P = 0.269). However, traps baited with SL-5 captured considerably more Cochliomyia macellaria (Fabricius), a closely related but nontarget fly., (Published by Oxford University Press on behalf of Entomological Society of America 2023.)

The study of insect fauna and their development and succession patterns on decaying cadavers is crucial to promoting insect evidence as a useful tool in forensic science, particularly for estimating the postmortem interval (PMI). Body decomposition and arthropod succession are affected by many factors and exhibit substantial regional variations; therefore, detailed succession studies in different biogeographic regions are required for understanding the successional patterns of insects in various environments. This study was conducted in the summer of 2021 using three domestic pig carcasses (Sus scrofa domestica L., 1758) in the field of Shizuishan City, Ningxia, Northwest China. A total of 40 species of necrophagous insects belonging to three orders and 16 families were collected. Among Diptera, Lucilia sericata (Meigen,1826), Chrysomya megacephala (Fabricius,1794), and Phormia regina (Meigen,1826) (Diptera: Calliphoridae) were the dominant species. As for Coleoptera, the dominant species changed throughout the process of carcass decomposition from Saprinus semipunctatus (Fabricius,1792) (Coleoptera: Histeridae) to Dermestes maculatus DeGeer,1774 and Dermestes frischii Kugelann,1792 (Coleoptera: Dermestidae). The carcasses desiccated rapidly and reached the remains stage under extreme conditions of high temperatures and low humidity, after which a large amount of dried tissue of the carcasses attracted populations of Coleoptera, particularly Dermstidae, which were abundant and remained until the end of the experiment on day 50. The current study is the first forensic entomological investigation of succession in Northwest China and provides basic data for the estimation of PMI during summer in this region., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The New World Screwworm, Cochliomyia hominivorax (Calliphoridae), is the most important myiasis-causing species in America. Screwworm myiasis is a zoonosis that can cause severe lesions in livestock, domesticated and wild animals, and occasionally in people. Beyond the sanitary problems associated with this species, these infestations negatively impact economic sectors, such as the cattle industry. Here, we present a chromosome-scale assembly of C. hominivorax's genome, organized in 6 chromosome-length and 515 unplaced scaffolds spanning 534 Mb. There was a clear correspondence between the D. melanogaster linkage groups A-E and the chromosomal-scale scaffolds. Chromosome quotient (CQ) analysis identified a single scaffold from the X chromosome that contains most of the orthologs of genes that are on the D. melanogaster fourth chromosome (linkage group F or dot chromosome). CQ analysis also identified potential X and Y unplaced scaffolds and genes. Y-linkage for selected regions was confirmed by PCR with male and female DNA. Some of the long chromosome-scale scaffolds include Y-linked sequences, suggesting misassembly of these regions. These resources will provide a basis for future studies aiming at understanding the biology and evolution of this devastating obligate parasite., (© The Author(s) 2022. Published by Oxford University Press on behalf of Kazusa DNA Research Institute.)

Blowflies (Diptera: Calliphoridae) are of great importance in forensic entomology and in determining the minimum post-mortem interval, as they may be the first group of insects to colonize decomposing remains. Reliable species identification is an essential prerequisite. Classically, morphological characters or DNA sequences are used for this purpose. However, depending on the species and the condition of the specimen, this can be difficult, e.g., in the case of empty fly puparia. Recent studies have shown that cuticular hydrocarbon (CHC) profiles are species-specific in necrophagous taxa and represent another promising tool for identification. However, the population-specific variability of these substances as a function of e.g., local climatic parameters has not yet been sufficiently investigated. The aim of this study was to determine the geographical variation of CHC profiles of the blowfly Calliphora vicina (Robineau-Desvoidy, 1830) depending on different countries of origin. Flies were reared in the United Kingdom, Germany, and Turkey in common garden experiments under ambient conditions. CHC profiles of the resulting adult flies and their empty puparia were analyzed using gas chromatography-mass spectrometry. Data were visualized by principal component analysis and clustered by population. The populations of the United Kingdom and Germany, both having similar climates and being geographically close to each other, showed greater similarities in CHC profiles. However, the CHC profile of the Turkish population, whose climate is significantly different from the other two populations, was very different. Our study confirms the high potential of CHC analysis in forensic entomology but highlights the need to investigate geographical variability in chemical profiles., (© Crown copyright 2022.)

This study investigates the impacts of three different heavy metals at different concentrations on some life-history traits of Lucilia sericata (Meigen 1826) (Diptera: Calliphoridae). First-instar larvae of L. sericata were reared on a diet containing four concentrations (0.25, 0.50, 1, and 2 µg/g) of the heavy metals (cadmium, zinc copper). The parameters measured were larval and pupal mortality, larval length, adult, pupal, and larval weight, and development time. Larval and pupal survival decreased as heavy metal concentrations increased. Pupal weight was significantly different among heavy metals andc oncentrations, but the adult weight was not significantly different among heavy metals and concentrations. The larval length was significantly different among concentrations and heavy metals. This study reveals that the presence of heavy metal in carrion or the carrion environment should be inconsidered when estimating the time of colonization in forensic investigations., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Forensic entomology is an important field of forensic science that utilizes insect evidence in criminal investigations. Blow flies (Diptera: Calliphoridae) are among the first colonizers of remains and are frequently used in determining the minimum postmortem interval (mPMI). Blow fly development is influenced by a variety of factors including temperature and feeding substrate. Unfortunately, dietary fat content remains an understudied factor in the development process, which is problematic given the high rates of obesity in the United States. To study the effects of fat content on blow flies we investigated survivorship, adult weight, and development time (overall and by sex) of Lucilia sericata (Meigen) and Phormia regina (Meigen) on ground beef with 10%, 20%, or 27% fat. As fat content increased, survivorship decreased across both species with significant impact to P. regina. While P. regina adults were generally larger than L. sericata across all fat levels, only L. sericata demonstrated a significant (P < 0.05) difference in weight by sex. Overall development times did not vary by fat content, excluding 27% for P. regina. Additionally, development times did not vary by sex for P. regina but did differ (P < 0.05) for L. sericata with females taking longer to develop. These findings provide insight into the effect of fat content on blow fly development, a factor that should be considered when estimating an mPMI. By understanding how fat levels affect the survivorship and development of the species studied here, we can begin improving the practice of insect evidence analysis in casework., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Forensic Entomology uses arthropods to aid in legal investigations. This study checked the biological response of Chrysomya putoria pupae to submersion in fresh water for up to 6 d, evaluating the critical submersion time, survival rate, and development time of the flies. Adults were collected using fish baits in two typical traps. Seven hundred and twenty fourth-generation pupae with 2 d of development were used and separated into submergence intervals: 24, 48, 72, 96, 120, and 144 h. An additional 120 pupae were used as a control. Each treatment was done in triplicate, consisting of 40 pupae distributed in four tulle-sealed test tubes containing 10 pupae each. All tubes of each treatment were co-adhered in test tube racks and were submerged in mineral water in a container with constant oxygenation, except those of the control group, which were not submerged. The tubes were removed from the water according to their respective submersion interval, until 144 h was completed. The control group had a survival rate of 90%, while the 24-h treatment had 85% and the 48-h treatment had 35.8%. The critical submersion time for pupae was 72 h, with 100% mortality by 144 h. The average development time for the control group was 3.2 d, while the 24- and 48-h treatments developed in 4.3 and 6.3 d, respectively. The longer the individuals were submerged, the lower the survival rate was, while the development time increased. The data obtained in this study have potential in applications to estimate the interval of submersion of a cadaver., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Maggot debridement therapy (MDT) is a therapy with the medical use of sterile fly larvae of certain species, particularly those within the Calliphoridae family including green bottle fly, Lucilia sericata (Meigan, Diptera: Calliphoridae), for treating chronically infected wounds and ulcers. Lucilia sericata flies were maintained under insectary conditions, and the eggs were sterilized using three treatments: hydrogen peroxide solutions, used as a hand disinfectant (Treatment 1-T1), hydrogen peroxide, surface disinfectant (Treatment 2-T2), and SaniHigene (Treatment 3-T3) and the control (without treatment). All three treatment caused the complete sterilization of eggs, and no bacterial colonies were found on the blood agar culture. The egg hatching rate after 72 h was much higher than after 24 h. Egg mortality in hydrogen peroxide solutions, T1 and T2, was 3-4% and less than in solution T3 (13%). Owing to less mortality and more sterility of the eggs, the aforementioned solutions are suggested to be appropriate for sterility in maggot therapy., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Many flies of Diptera are common entomological evidence employed in forensic investigation. Exploring the existence of inter- and intra-species genomic differences of forensically relevant insects is of great importance. Aldrichina grahami is a common blow fly species of forensic importance. The present study characterized the gene repertoires of A. grahami, and provides insights into issues related to forensic entomology, such as necrophagous behavior, gene family features, and developmental patterns. Gene families were clustered and classified according to their function in different aspects of the necrophagous lifestyle, generating several gene repertoires. The genes under positive selection pressure and evolutionary changes were screen and identified. Moreover, genes that exhibited potential prediction value in the post mortem interval (PMI) estimation and development of immature stages were subjected to analysis based on the developmental transcriptome. Related insect species were compared at the genomic level to reveal the genes associated with necrophagous behaviors. The expression of selected genes in separated repositories was verified using qPCR. This work was conducted using a high-quality chromosome-level genome assembly of A. grahami and its developmental transcriptome. The findings will facilitate future research on A. grahami and the other forensically important species., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The New World screwworm, Cochliomyia hominivorax (Coquerel 1858) (Diptera: Calliphoridae), is a serious parasite of livestock, humans, and other warm-blooded animals. It has been eradicated from the northern parts of its historical range down to the Panama-Colombian border where a permanent barrier zone is maintained. This eradication was accomplished through using the sterile insect technique (SIT). In 2016 there was an outbreak of C. hominivorax in the Florida Keys. In only six months, this pest was successfully re-eradicated using SIT, but the geographic origin of the invasion has yet to be resolved. It was previously determined that the Florida flies most likely represented a single invasion, and it was recommended that a finer-scale genetic assessment should be completed. Thus, this current proof-of-concept study aimed to develop a population genetic database using single nucleotide polymorphisms (SNPs) to reference outbreaks and potentially identify the origin of the Florida outbreak. This initial database consists of wild-caught samples from 4 geographic locations as well as laboratory colony samples that originated from 7 additional locations using a genotyping by sequencing (GBS) approach. Geographic population structuring was identified for twelve populations that clustered according to geographic location. The Florida outbreak samples appeared similar to samples from the outer Caribbean cluster which included samples from Dominican Republic and Trinidad and Tobago, however, these results will be further clarified with the replacement of laboratory colony samples with future wild-caught samples., (Published by Oxford University Press on behalf of Entomological Society of America 2022.)

The present study aimed to determine the effects of the soil type and the moisture contents on the some life-history parameters of Lucilia sericata (Meigen, 1826) (Diptera: Calliphoridae). The larval and pupal survival, duration of development periods, and the weight of pupae and adult of L. sericata were examined at three different types of soil (clay, loamy, and sandy) with five moisture contents (0, 25, 50, 75, 100%). The post feeding L. sericata larvae were transferred to a plastic cup filled with soils with different moisture content, and all cups were kept at 27°C, 65% RH, and a photoperiod of 12:12 (L:D) h. In this article, the effects on some life-history parameters were compared with a two-way analysis of variance (ANOVA). Accordingly, the findings of this article indicate that moisture contents of soils and soil type have a significant effect on the development time of L. sericata. Furthermore, it has been found that larvae can survive on extremely wet substrates (75-100% humidity), although their development time is longer than other moisture content. Pupal and adult weight was significantly different among soil type and soil moisture. Our results provide a reference information for sufficient estimation of the effects of changes in moisture-related to soil type or climate on studies of forensic and biological control of L. sericata., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Calliphora grahami (Aldrich, 1930) (Diptera: calliphoridae) is a forensically important blow fly that is widely distributed across Asia, North America, Russia, and Mexico. Calliphora grahami is frequently found on corpses during the spring, autumn, and winter seasons. It is among the early colonizers of cadavers during the cold season, and sometimes, the only necrophagous blow fly on cadavers. Therefore, this species is of forensic significance, although very few studies have explored the application of its intrapuparial age for PMI estimation. This study aimed to examine the intrapuparial development of C. grahami and establish a method for estimating its intrapuparial age. Herein, the C. grahami puparia were studied under six different temperatures (13, 16, 19, 22, 25, and 28°C) for the intrapuparial age estimation, and a total of 5776 puparia were sampled. The morphological changes were divided into 11 stages based on the 1) development of legs and wings, 2) differentiation of the head, thorax, and abdomen, 3) growth and color of the bristles, and 4) color changes of the compound eyes. The corresponding time of each stage was determined. Moreover, the observation and classification of individual morphological features, including compound eyes, antennae, mouthparts, thorax, abdomen, legs, and wings was used to improve the precision of intrapuparial age estimation. The findings of this study provide important information on the use of C. grahami intrapuparia to estimate the minimum postmortem interval (PMImin)., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

This is the first study in Brazil that monitored the nocturnal oviposition behavior of Chrysomya putoria, a species of forensic importance, in order to verify if individuals of this species oviposit at night. Groups of 10 flies (5 male and 5 female) distributed in ten cages were kept in a fume hood and submitted to total darkness or exposed to artificial light for eleven consecutive hours through four experimental conditions in the laboratory. Two verifications were made to see if the females oviposited in the offered substrate of about 20 g of chicken gizzard per cage. Verification 1 occurred at 09:00 pm in the evening experiments and at 09:00 am in the daytime experiments. Verification 2 occurred at 05:00 am in the night experiments and at 05:00 pm in the daytime experiments. Each experiment lasted 5 d. C. putoria laid eggs at night (with or without light) and the quantity of eggs was significantly similar to those produced during the day under natural light or in total darkness. Only the amount of eggs produced during the day in the absence of light was considerably greater than in the typical daytime period. The females oviposited in greater quantity in the nights when the average temperature was between 23 and 24.8°C and relative humidity above 81%. Ovipositions only occurred at temperatures above 21°C and humidity above 56% during the day. Finally, it is necessary that more evaluations on the effect of variables on blowfly behavior are performed to better understand nocturnal oviposition., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Studies under constant temperatures are the most common to estimate the Postmortem Interval (PMI). It is imperative that forensic sciences have data from studies carried out in the field. Therefore, this work aims to: (1) evaluate the parameters (weight, length, development time) associated with the life cycles of Lucilia ochricornis (Wiedemann) (Diptera: Calliphoridae) and Lucilia purpurascens (Walker) under experimental conditions in the field considering fluctuating temperatures, and (2) compare these results with those known and published by the same authors for cultures realized in the laboratory under constant temperatures; which will permit us to contrast the most widely used existing methodologies for forensic application in estimating the minimum postmortem interval (PMImin). For each season of the year, cultures of both species were made in the field, collecting information on temperature, humidity, and photoperiod to perform laboratory cultures, later comparing: development time, length, weight, and Accumulated Degree-Hours (ADH) in both types of cultures. Methods for estimating the PMI were obtained and validated with the information of the cultures grown in the field. The two types of cultures showed differences between each other for both species. The forensic use methods to estimate PMI were enhanced and their precision increased when maximum larval length data were used, and it was also concluded that feeding larval stages are the most accurate to be used in making estimates because the larva is growing. The estimation of the PMI through the use of necrophagous flies development remains reliable for obtaining the PMImin., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Adult Calliphoridae flies, as well as their immature stages collected from carcasses, have been used as evidence in forensic investigations to estimate the postmortem interval (PMI), particularly those of the genus Chrysomya as it is one of the first genera to colonize a corpse. Chrysomya rufifacies (Macquart 1842), due to its appearance in cadaveric remains, plays a fundamental role in the study of forensic entomology. For this reason, we determined the biological cycle of C. rufifacies under semicontrolled laboratory conditions: uncontrolled average fluctuating temperature of 29. 76 ± 3. 22°C, uncontrolled average fluctuating humidity of 48. 91 ± 11.13%, and a controlled photoperiod of 12/12 (L/O). We established that the total development time from oviposition to adult emergence of C. rufifacies was 6. 5 d. The eggs took 12 h to hatch after oviposition. The complete larval stage took 60 h (instar 1 = 12 h, instar 2 = 12 h, instar 3 = 24 h, instar 3 post-feeding = 12 h). The pupa had a duration of 84 h. The species needed a total of 4642.8(±4.59) accumulated degree-hours (ADH) to complete its biological cycle., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America.All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Entomotoxicology is an emerging science in the forensic investigation of death; it now influences judicial activities. Briefly, carrion flies ingest various chemicals as they feed on a cadaver. In entomotoxicology, such toxicological compounds are qualitatively and quantitatively analyzed in insects recovered from carrion. Some of these chemicals can disrupt flies' cellular physiology, leading to acceleration or delay in their developmental rate. In this study, the effect of zolpidem-contaminated tissue on the dipteran flies Chrysomya megacephala (Fabricius 1794) and Chrysomya saffranea (Bigot 1877) was assessed. Zolpidem tartrate is a sedative-hypnotic compound commonly used to treat anxiety and sleeping disorders, but it is associated with death from suicide. The effects were tested by providing the flies with liver tissue treated with various concentrations of zolpidem tartrate. The morphological parameters and development rate for all life cycle stages of treated insects were determined and used to estimate postmortem intervals. Results showed that the morphological parameters, i.e., weight, width, and length and rate of development of C. megacephala and C. saffranea was negatively associated with zolpidem tartrate concentration of C. saffranea, were negatively associated with the concentration of zolpidem tartrate, indicating that arthropod evidence evaluation can help establish the cause and time of death., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Leishmaniasis is a neglected tropical disease that affects millions of people around the world. Larval excretion/secretion (ES) of the larvae of flies of the Calliphoridae family has microbicidal activity against Gram-positive and Gram-negative bacteria, in addition to some species of Leishmania. Our study aimed at assessing the in vitro efficacy of Lucilia cuprina larval ES against the promastigote and amastigote forms of Leishmania amazonensis, elucidating possible microbicidal mechanisms and routes of death involved. Larval ES was able to inhibit the viability of L. amazonensis at all concentrations, induce morphological and ultrastructural changes in the parasite, retraction of the cell body, roughness of the cytoplasmic membrane, leakage of intracellular content, ROS production increase, induction of membrane depolarization and mitochondrial swelling, the formation of cytoplasmic lipid droplets and phosphatidylserine exposure, thus indicating the possibility of apoptosis-like death. To verify the efficacy of larval ES on amastigote forms, we performed a phagocytic assay, measurement of total ROS and NO. Treatment using larval ES reduced the percentage of infection and the number of amastigotes per macrophage of lineage J774A.1 at all concentrations, increasing the production of ROS and TNF-α, thus indicating possible pro-inflammatory immunomodulation and oxidative damage. Therefore, treatment using larval ES is effective at inducing the death of promastigotes and amastigotes of L. amazonensis even at low concentrations., (© The Author(s) 2021. Published by Oxford University Press on behalf of FEMS. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Larval therapy (LT) is a therapeutic modality that uses larvae of necrophagous flies for the treatment of wounds. The use of this therapy presents several benefits, due to the action of the larvae that remove necrotic tissue selectively, exercising antimicrobial action, and promoting healing. There are situations in which LT proves to be the only or the best alternative for the patient, such as wounds infected by multidrug-resistant microorganisms or when treatment difficulties may lead to an indication for amputation. The purpose of this study was to compare the efficiency of LT, using larvae of Chrysomya megacephala, with that of antibiotic therapy in the treatment of cutaneous wounds infected with Pseudomonas aeruginosa. Twenty-four rabbits were used to perform the experiment, distributed in four groups: group 1, induced wound without bacterial infection; group 2, induced wound with bacterial infection; group 3, induced wound with bacterial infection and antibiotic therapy; group 4, induced wound with bacterial infection and LT. The macroscopic, microscopic, and statistical analyses indicated that LT was as effective as antibiotic therapy in wound healing., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Chrysomya megacephala (Fab. 1794) (Diptera: Calliphoridae) is a very important species for forensic entomology, mainly contributing estimations of the postmortem interval (PMI) in judicial investigations. There are some doubts about the nocturnal oviposition of these flies, which could lead to errors in the PMI calculation. This study aimed to monitor the nocturnal oviposition behavior of this species through four experimental conditions carried out in laboratory. Ten cages, each containing five males and females (n = 100), were kept in a fume hood and subjected to total darkness or to artificial light for 11 consecutive hours. Two verifications were performed to determine whether the females deposited eggs on the substrate of ~20 g of chicken gizzards per cage. The first verification occurred at 9:00 pm in nocturnal experiments and at 09:00 am in diurnal experiments. The second verification occurred at 05:00 am in nocturnal experiments and at 05:00 pm in diurnal experiments. Each experiment lasted 5 d. Chrysomya megacephala deposited eggs at night under artificial light and in total darkness, but the amount of eggs was significantly lower when compared with the daytime experiments in dark conditions and under natural light. Oviposition occurred when the average temperature was around 25°C (± 2°C) and relative humidity around 73% (± 6%). Night oviposition is a possibility which should not be ruled out. Thus, future experiments are recommended., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Avoiding competition is thought to explain insect successional patterns on carrion, but few studies have looked at competition directly. We use replacement series experiments with three species of blow flies: Phormia regina (Meigen) (Diptera: Calliphoridae), Lucilia sericata (Meigen) (Diptera:Calliphoridae), and Chrysomya rufifacies (Macquart) (Diptera:Calliphoridae) to characterize competitive relationships. From experimental results, P. regina showed a significant competitive advantage over L. sericata. Infestation of carrion differs between L. sericata and P. regina; specifically, L. sericata oviposits on carrion without any delay, while P. regina typically delays oviposition. Our findings are consistent with the notion that differences in oviposition times represent a mechanism for L. sericata to avoid potential competition. Competition by C. rufifacies differs since C. rufifacies, in the event of a limited food supply, will prey on other maggot species. In replacement series experiments, C. rufifacies killed all P. regina in mixed treatments, representing an ultimate competitive advantage. In the United States, these two species do not often overlap because of differences in seasonal distribution. However, with climate change, phenological separation may grow less distinct. Surprisingly, in replacement series experiments with C. rufifacies and L. sericata, no competitive interactions were observed. In other studies, L. sericata has been shown to form clusters away from predaceous maggots, allowing improved survival, which may account for the absence of predation by C. rufifacies. Finally, this study shows that replacement series models are useful in measuring competition, supporting the notion that interspecific competition between necrophagous insect species may have driven life history traits of those species., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Diptera Mesembrinellidae present great ecological importance as potential bioindicators and Calliphoridae stand out for their medical-sanitary importance as potential vectors of pathogens and causes of human and animal diseases such as myiasis. They beneficially aid in elucidating crimes in forensic entomology, contributing to determine the postmortem interval (PMI) and assisting in the healing process of necrotic wounds in larval therapy. This study aimed to inventory the Calliphoridae and Mesembrinellidae fauna in order to evaluate structural parameters and to apply faunal indexes in catching species in the Itaipu-Piratininga lagoon complex, in Niterói (Rio de Janeiro), Brazil. The collections were carried out monthly from September 2015 to August 2016 with fish bait (sardines) exposed for 48 h, totaling six traps installed in three physiognomies (mangrove, ombrophilous forest, and restinga). In total, 9,773 individuals were captured, divided into two families, five genera, and 11 species. Chrysomya megacephala (Fabricius, 1794) was the predominant species, while Mesembrinella bellardiana (Aldrich, 1922) was the least representative species, indicating the preservation level of this ecosystem. Ombrophilous forest was the physiognomy with greater diversity and restinga was the physiognomy with greater species dominance. The similarity index indicated greater similarity among the mangrove and restinga populations., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America.All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Calliphoridae and Mesembrinellidae are relevant to environmental conservation, public health, and forensic entomology. Researches regarding the flight behavior and the influence of abiotic factors on these insects may assist the application of entomology sciences. This study aimed to analyze the population fluctuation of Calliphoridae and Mesembrinellidae, verifying the influence of environmental factors, trap height, and the anthropic effect in the Itaipu-Piratininga lagoon complex, Niterói (RJ). The collections were carried out monthly from September 2015 to August 2016, with fish bait (sardines) exposed for 48 h, totaling six traps, installed in three physiognomies (mangrove, ombrophilous forest, and restinga) at 1.5 and 2.5 m from the ground. Nine thousand seven hundred seventy-three individuals were captured, comprising two families, five genera and 11 species. Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae) was the predominant species and Mesembrinella bellardiana (Aldrich, 1922) (Diptera: Mesembrinellidae) the least representative, indicating the low preservation level of this ecosystem. There was a weak, positive correlation between abundance of Chloroprocta idioidea (Robineau Desvoidy, 1830) (Diptera: Calliphoridae), Ch. megacephala, Chrysomya putoria (Wiedemann, 1818) (Diptera: Calliphoridae), and Lucilia cuprina (Wiedemann, 1830) (Diptera: Calliphoridae) with the temperature, as well as between the abundance of C. idioidea and Chrysomya albiceps (Wiedemann, 1819) (Diptera: Calliphoridae) with precipitation; however, there was no correlation between abundance and relative humidity. No significant influence of the trap height was observed. We stated a high influence of anthropic effects on the restinga and mangrove physiognomies, while the forest physiognomy still retains its preserved characteristics, with the dominance of forestall species., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America.All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Forensic entomology is a well-established tool for evaluating the death, or abuse, of a person. Insect evidence provides valuable information as related to time of colonization and movement of remains from one location to another. Blow flies (Diptera: Calliphoridae) are commonly found on human remains throughout most stages of decomposition, and when entomological evidence is collected these taxa tend to be the most numerous. However, very few crime laboratories across the country have collection and rearing protocols for these forensically important insects. A lack of knowledge in collection techniques and limited access to an appropriate food source are the main reasons for absence in adequate collection and rearing protocols. Thus, when crime scene investigators or pathologists collect insects, they are often mishandled (e.g., placed into containers with no air holes, no food, or a food source that is not sustainable for their development). To address this issue, we analyzed easily accessible and cost-efficient alternative food sources for blow flies; specifically, tuna and wet cat food compared to beef liver (control). Survivorship and development were examined for each food source using the blow flies Phormia regina (Meigen) (Diptera: Calliphoridae) and Cochliomyia macellaria (Fabricus) (Diptera: Calliphoridae). These findings provide an overview of possible alternatives that could be used as a sustainable food source in crime laboratories when immediate action from a forensic entomologist cannot be obtained., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The attraction of necrophagous insects, particularly blow flies, to corpses and carrion is of ecological, economic, and agricultural importance, although the mechanisms by which it occurs are not well understood. Much of the published research on blow fly attractants has focused on volatiles emitted from carrion surrogates, but little attention has been given to the possibility that blow fly eggs themselves may emit chemical cues that are responsible for conspecific and heterospecific insect attraction. In this study, the headspace volatiles emitted from eggs representing two aggregated oviposition events that were collected 1 mo apart from two species of the Calliphoridae family (Order: Diptera), Lucilia sericata (Meigen), and Phormia regina (Meigen) were analyzed via solid-phase microextraction-facilitated GC-MS. The volatiles' profiles were found to be consistent between samples representing the same species, but unique between the two species. Over 100 molecules covering a wide range of compound classes that included alcohols, aldehydes, esters, amines, ketones, and organosulfur compounds were identified. The profile of volatiles emitted from the L. sericata eggs contained several alkanes and aldehydes, whereas salient features of the P. regina headspace included numerous esters and ketones. Between the two species, 42 compounds were shared, several of which were carboxylic acids. Little overlap between the range of compounds detected and those reported to be emitted from decomposing remains was observed., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Necrophagous insects play an important role in the decomposition of vertebrate carrion. The documented colonization, development, and succession of blow flies (Diptera: Calliphoridae) and other arthropods on decomposing carcasses make their communities relevant for use in decomposition ecology and forensic investigations. This relevance relies on the local pool of species available to colonize a carcass, but such community level survey data are not always available. The objective of this research was to conduct a baseline survey of adult Calliphoridae communities from urban-rural land use types in the Great Lakes region. To test how adult blow fly distribution varies with changing landscape in Mid-Michigan, sampling with baited jars and hanging traps was implemented over the summer months of June, July, and August 2017. To determine how blow fly communities differed in urban to rural land cover, seven cities were selected with site locations ranging from high intensity developed areas to cultivated crop fields. Over 97,000 individual flies were captured represented by 11 Calliphoridae species. The adult Calliphoridae communities were primarily structured by land use type and month of collection, with these two factors interacting, suggesting that the effect of location varied by time of year. The two most abundant species, Phormia regina (Meigen) and Lucilia sericata (Meigen), cumulatively comprised 88.5% adult flies from Mid-Michigan. These findings provide a baseline database of Great Lakes Calliphoridae, with potential use in forensic research and casework., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Insect age estimates can be useful for estimating the postmortem interval when certain assumptions are met. Such estimates are based on species-specific development data that are temperature-dependent and variable, and therefore prone to different degrees of error depending on the combination of data sets, calculations, and assumptions applied in a specific instance. Because of this potential error, validating the methods employed is necessary for determining accuracy and precision of a given technique. For forensic entomology, validation of development data sets is one approach for identifying the uncertainty associated with insect age estimates. Cochliomyia macellaria (Fabricius) is a primary colonizer of remains across the United States and is commonly encountered in forensic investigations. A development study for this species was produced for a central Texas, U.S. population; the variation associated with this data set and the pre-appearance interval were previously explored in an ecological model. The objectives of this study were to determine the accuracy of the development data and the validity of the ecological model when applied to immatures of known age developing under field conditions. Results indicate this data set is an accurate predictor of insect age when using development stage, supporting the validity of the ecological model in central Texas. Age predictions made with all stages present in a sample were more accurate than predictions made with the most developed stage in a sample, and estimates of age when using the prepupal stage were overestimated regardless of prediction method, though thermal requirements for total development were similar., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The inhibitory effect of Chrysomya rufifacies (Macquart) and Cochliomyia macellaria (Fabricius) larval excretions-secretions (ES) on Staphylococcus aureus was determined using a portable colorimetric method without the need for any dedicated spectral instrument. Polystyrene 96 well microplates were used and 100 μl of the bacterial inoculum (5 × 105 CFU/ml) plus 100 μl of the dipteran exosecretions at different concentrations were added to each well. Subsequently, 50 μl of a 1% solution of the triphenyl tetrazolium chloride stain was added to each well to determine the bacterial viability. The color development in each well was measured with the ImageJ software S. aureus was exposed to different concentrations of the ES of both species individually. At a concentration of 800 ppm ES of C. rufifacies or Co. macellaria, bacterial growth was inhibited 97.45 ± 1.70% and 82.21 ± 1.88%, respectively. As expected, exposure to a lower concentration (i.e., 50 ppm) was less inhibitory (C. rufifacies ES, 77.65 ± 4.25% and Co. macellaria ES, 43.54 ± 4.63%). This study demonstrates for the first time the bactericidal activity of C. rufifacies and Co. macellaria ES against S. aureus. This finding is promising as it could result in the identification and synthesis of proteins capable of suppressing pathogen development in wounds. Additionally, the proposed method can simplify the use of expensive laboratory instruments for antimicrobial activity determination., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Blow fly (Diptera: Calliphoridae) larvae are commonly used in forensic cases to determine postmortem intervals using development rates and successional changes in community composition. Studies are conducted from different regions to provide these data. We wanted to know how widely applicable these data are. We examined whether urbanized landscapes have distinct urban blow fly communities or whether the community composition in urbanized areas is simply a variation of that found in the surrounding habitat or ecozone. Using liver baited traps, we sampled 7,272 flies from 32 sites across Canada and used mapping analysis to assess urban and rural landcover classifications, and compared urban and rural species abundance and composition. Blow fly species communities from urban areas across Canada were made up of similar species and differed from the communities found in nearby rural sites. Trapping at rural sites caught more blow flies compared with urban sites (mean flies/site 59.5 and 12.4). Of the 14 species caught, 8 were caught at urban sites, 61% of these being Cynomya cadaverina Robineau-Desvoidy, 14% Phormia regina Meigen, and 11% Lucilia sericata (Meigen). In rural sites, all 14 species were caught, 41% of specimens caught were P. regina, 21% C. cadaverina, 10% Calliphora vomitoria (Linnaeus), with only 4% L. sericata. These data suggest that regional studies are appropriate for forensic entomology applications in urban landscapes, given the similar trends across Canada, less so for wilderness or rural landscapes., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Determining the age of an insect collected in a corpse is a key element to estimate the minimum postmortem interval in forensic entomology. Along with models of accumulated degree-hours, the estimation of the age based on larval weight and length is among the most common methods for this purpose. Sarconesia chlorogaster (Wiedemann, 1830), along with other Calliphoridae, is an important species in forensics in South America. However, studies analyzing the weight and length of S. chlorogaster larvae during development have not yet been done. In this work, we develop growth models based on the weight and length of larvae in different temperatures and provide formulas that can be used to estimate the age of the larvae based on the weight and length for each temperature. Larvae were reared at six temperatures (10, 15, 20, 25, 30, 35°C), and the measurements of larval length and weight were taken during all development until they stopped feeding. For the measurements, we attempted to use methodologies that can be easily replicated and do not require equipment that could be of difficult access. The results of this work come to complement and improve the use of S. chlorogaster in forensic applications., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The medico-legal section of forensic entomology focuses on the analysis of insects associated with a corpse. Such insects are identified, and their life history characteristics are evaluated to provide information related to the corpse, such as postmortem interval and time of colonization. Forensically important insects are commonly identified using dichotomous keys, which rely on morphological characteristics. Morphological identifications can pose a challenge as local keys are not always available and can be difficult to use, especially when identifying juvenile stages. If a specimen is damaged, certain keys cannot be used for identification. In contrast, molecular identification can be a better instrument to identify forensically important insects, regardless of life stage or specimen completeness. Despite more than 20 yr since the first use of molecular data for the identification of forensic insects, there is little overlap in gene selection or phylogenetic methodology among studies, and this inconsistency reduces efficiency. Several methods such as genetic distance, reciprocal monophyly, or character-based methods have been implemented in forensic identification studies. It can be difficult to compare the results of studies that employ these different methods. Here we present a comprehensive review of the published results for the molecular identification of Diptera of forensic interest, with an emphasis on evaluating variation among studies in gene selection and phylogenetic methodology., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Aging blow fly eggs can be critical to a forensic investigation, but there are currently no forensically useful timelines describing internal anatomical changes in embryological development. This is partly due to the lack of an economical, rapid and technically simple histological technique to allow mass production of slides for research and casework. We present a histological method that uses a slightly modified standard laboratory processing run with 1 h fixation in 10% formalin, 2 h softening in Molliflex and Haemotoxylin and Eosin (H&E) staining. We also present a summary of the internal anatomical changes that can be visualized using our technique in the developing eggs of Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae). We examined eggs from at least three different females grown at 15 °C and sampled at 6 h intervals, and eggs grown at 20 °C and sampled at 3 h intervals. Blind aging trials demonstrated that it is possible to accurately age material grown at 20 °C to within 6 h (but attempts to further narrow this interval resulted in errors in one-third of cases). It was also possible to see sufficient anatomical detail to age eggs preserved for forensic casework 5, 8, 9, 10 and 11 years previously. Additionally, we determined that section quality was improved by 5 s fixation in hot water prior to preservation in ethanol. However, hot water fixation for longer than this increased the level of section artefact.

Common DNA-based species determination methods fail to distinguish some blow flies in the forensically and medically important genus Lucilia Robineau-Desvoidy. This is a practical problem, and it has also been interpreted as casting doubt on the validity of some morphologically defined species. An example is Lucilia illustris and L. caesar , which co-occur in Europe whilst only L. illustris has been collected in North America. Reports that these species shared both mitochondrial and nuclear gene sequences, along with claims that diagnostic morphological characters are difficult to interpret, were used to question their separate species status. We report here that amplified fragment length polymorphism profiles strongly support the validity of both species based on both assignment and phylogenetic analysis, and that traditional identification criteria based on male and female genital morphology are more reliable than has been claimed.

Large numbers of flies are needed to produce the quantity of larvae required for insect bioconversion of waste. However, this 'mass-rearing' may negatively affect adult survival and reproductive output. This study assessed the suitability for mass-rearing of four blow fly species, Chrysomya chloropyga, Chrysomya chloropyga (Wiedemann), Chrysomya megacephala (F.), Chrysomya putoria (Wiedemann) and Lucilia sericata (Meigen). Flies were kept at densities of 20, 50, 100, 250, 500, and 1,000 flies per 30 × 30 × 30 cm cage with an even sex ratio. Time to 50% mortality (LT50) was recorded, and the effects of density, species, and sex on LT50, fecundity, and fertility were determined. Females survived longer than males across all species. There was evidence for a trade-off between survival and high fecundity in L. sericata and C. chloropyga at density 250. C. megacephala had low fecundity across all densities. At high densities, C. putoria had the lowest mortality and highest fecundity, making it the most suitable for mass-rearing., (© The Author(s) 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

For all species, abiotic factors directly affect performance, survival and reproduction, and consequently, their geographic distribution. Species distribution models (SDMs) are important tools to predict the influence of abiotic factors in species distributions and has been more applied over the years. However, these models can be built under different algorithms and using different methods to select environmental predictors, which can lead to different results. Five different algorithms and two sets of environmental predictors were compared to predict the geographic distribution of the blowfly Sarconesia chlorogaster (Wiedemann) (Diptera: Calliphoridae). This species has several occurrence points and a considerable amount of biological data available, which makes S. chlorogaster a good model system to compare environmental predictors. Two sets of environmental predictors (mainly derived from temperature and humidity) were built, and the set based on the influence of abiotic variables on the ecophysiology of S. chlorogaster showed better results than the principal component analysis (PCA) approach using 19 climatic variables. We also employed five modeling algorithms-Envelope Score, Mahalanobis Distance, GARP, Support Vector Machines, and Maxent-and the latter two showed the best performances. The results indicate that temperature is the main factor shaping geographic distribution of S. chlorogaster through its effect on fitness. Furthermore, we showed that this species is mainly distributed in south, southeastern, and some northwestern and southwestern sites of South America. In addition, our results also predicted suitable areas in Ecuador and Colombia, countries without previous records., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Over the past several decades, there have been several strident exchanges regarding whether forensic entomologists estimate the postmortem interval (PMI), minimum PMI, or something else. During that time, there has been a proliferation of terminology reflecting this concern regarding "what we do." This has been a frustrating conversation for some in the community because much of this debate appears to be centered on what assumptions are acknowledged directly and which are embedded within a list of assumptions (or ignored altogether) in the literature and in case reports. An additional component of the conversation centers on a concern that moving away from the use of certain terminology like PMI acknowledges limitations and problems that would make the application of entomology appear less useful in court-a problem for lawyers, but one that should not be problematic for scientists in the forensic entomology community, as uncertainty is part of science that should and can be presented effectively in the courtroom (e.g., population genetic concepts in forensics). Unfortunately, a consequence of the way this conversation is conducted is that even as all involved in the debate acknowledge the concerns of their colleagues, parties continue to talk past one another advocating their preferred terminology. Progress will not be made until the community recognizes that all of the terms under consideration take the form of null hypothesis statements and that thinking about "what we do" as a null hypothesis has useful legal and scientific ramifications that transcend arguments over the usage of preferred terminology., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Commercial livestock facilities are faced with the challenge of managing large amounts of waste including manure and animal mortalities. One method of disposing of dead animals is composting. The cadavers are enveloped in carbon material that creates a barrier between the dead tissue and the surrounding environment. Dead tissue can release materials that not only contaminate the soil but also the groundwater and nearby surface water. Animal cadaver composting is designed to facilitate decomposition without the aid of carrion-feeding insects and reduce the presence of common pathogens associated with animal waste and dead tissue. The aim of this study was to evaluate insect activity associated with composted and exposed beef cadavers, specifically filth flies that can serve as mechanical vectors of important human pathogens such as E. coli 0157:H7. Greater numbers of all types of arthropods were trapped overall at the exposed animal site than the composted animal site. Most importantly, the number of filth flies was significantly lower at the composted site (P = 0.0009). Laboratory analysis of volatile organic compounds from composted and noncomposted rats indicated that known fly attractants such as dimethyl disulfide may be inhibited by the composting process. Implementing composting programs at livestock facilities could reduce the risk of flies spreading harmful pathogens to surrounding areas, including farms that grow fresh produce., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Forensic entomologists often rely on development data associated with a given species to estimate when it colonized human or other vertebrate remains. In most instances, these development studies are based on single species reared in isolation in the laboratory. This study examined the impact of excretions and secretions (ES) associated with third-instar Chrysomya rufifacies (Macquart), a predator, on the development of its prey, Cochliomyia macellaria (F.). Not surprisingly, Ch. rufifacies ES did not impact the development of first- or second-instar C. macellaria, which are typically not preyed on by Ch. rufifacies. However, development of third-instar C. macellaria, which do experience predation, was impacted. First, larvae were longer than those in the control (deionized water, dH2O). Filtering the ES and removing the associated bacteria and byproducts >0.2 µm dampened the previous impact observed by the unfiltered ES on third-instar C. macellaria. Second, third-instar C. macellaria treated with unfiltered ES completed pupariation 8 h quicker than the controls. Filtering the ES lessened this effect by 50%. And finally, third-instar C. macellaria treated with filtered or unfiltered Ch. rufifacies ES reached adulthood ∼5 h faster than controls treated with dH2O. In summary, these data have large ramifications for forensic entomology, as multiple species being present on decomposing remains is not uncommon. Understanding the impact of associated ES produced by interspecific cohorts on associated development could lead to more precise estimates of the minimum postmortem interval for forensic investigation of decomposing remains., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Chrysomya megacephala (Fabricius) is a forensically important fly that is found throughout the tropics and subtropics. We calculated the accumulated development time and transition points for each life stage from eclosion to adult emergence at five constant temperatures: 15, 20, 25, 30, and 35 °C. For each transition, the 10th, 50th, and 90th percentiles were calculated with a logistic linear model. The mean transition times and % survivorship were determined directly from the raw laboratory data. Development times of C. megacephala were compared with that of two other closely related species, Chrysomya rufifacies (Macquart) and Phormia regina (Meigen). Ambient and larval mass temperatures were collected from field studies conducted from 2001-2004. Field study data indicated that adult fly activity was reduced at lower ambient temperatures, but once a larval mass was established, heat generation occurred. These development times and durations can be used for estimation of a postmortem interval (PMI)., (© The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)