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Pyrethrum is a botanical insecticide derived from pyrethrum flowers. Feeding deterrence caused by pyrethrum has been reported in several sucking insects; however, there is no account of the cause of deterrence—whether from a single component or the combination of six active ingredients, called pyrethrins. We determined the feeding deterrence of natural pyrethrins, their two main components (pyrethrins I and II), and pyrethroid insecticides on the blowfly, Phormia regina. In a dual-choice feeding assay that minimized tarsal contact with food sources but allowed feeding through proboscises, natural pyrethrins, synthetic pyrethrins I/II, and allethrin were observed to induce deterrence at a concentration 16 times lower than the lowest concentration at which the knockdown rate increased. Feeding bouts were interrupted by intensive grooming of the proboscis at the deterring concentration, but no such grooming was observed to occur while feeding on the unpalatable tastants—NaCl, quinine, and tartaric acid. The underlying mode of action for the feeding deterrence of pyrethrins at sub-lethal concentrations probably occurs on the fly oral gustatory system, while differing from that of unpalatable tastants. The potent feeding deterrence of pyrethrins may provide effective protection for pyrethrum plants by rapidly deterring insects from feeding, before insecticidal activities occur.

Phormia regina (Meigen, 1826; Diptera: Calliphoridae) is a Holarctic species that rapidly colonizes carcasses and has been used as an indicator for determining the minimum postmortem interval. However, studies using morphological methods to estimate the intrapuparial age of P. regina are lacking. In this study, morphological changes within the puparium were observed under a stereomicroscope at 7 constant temperatures ranging from 16 °C to 34 °C.The intrapuparial period was categorized into 12 substages. Morphological indicators, including compound eyes, mouthparts, antennae, thorax, legs, wings, and abdomen, were recorded in detail. The observed morphological changes were divided into 6–10 substages, and the duration of each substage was also recorded in detail.The results of this study provide primary data for using the intrapuparial morphology of P. regina when pupae are collected at a crime scene and estimating the minimum postmortem interval.

Phormia regina (Meigen, 1826) (Diptera: Calliphoridae) can colonize carcasses quickly, and its immature stages are reliable entomological evidence for the estimation of the minimum postmortem interval (PMImin). There are discrepancies in the developmental data from previous studies on P. regina, and the related PMImin indicators need to be refined. We investigated the accuracy of forensic entomological evidence using development durations, growth accumulated degree hours, and larval body length variations of P. regina at seven constant temperatures ranging from 16 to 34 °C. We also established development models such as the isomorphen diagram, thermal summation model, isomegalen diagram, and body length simulation equation to assist with PMImin estimation. The developmental duration of P. regina from egg to adult at 16, 19, 22, 25, 28, 31, and 34 °C was 840.8 ± 42.8 h, 580.1 ± 10.1 h, 390.4 ± 8.7 h, 316.8 ± 9.4 h, 291.4 ± 21.2 h, 238.4 ± 2.8 h, and 222.5 ± 5.2 h, respectively. The lower threshold temperature TL was 9.97 ± 0.50 °C, while the thermal constant K was 5052.7 ± 229 degree days. The lower developmental thresholds, intrinsic optimum temperature, and upper lethal developmental threshold obtained by the Optim SSI models were 13.15, 21.20, and 36.86 °C, respectively. This study aims to provide developmental models for P. regina aimed at common case-site temperatures in the northern provinces of China, which can be used for accurate PMImin estimation.

•The European necrophagous blow flies of forensic importance can be successfully identified using wing measurements.•Wing measurements are a promising alternative to other methods of identification of necrophagous Calliphoridae.•This method can be particularly useful for identification of large samples of females collected during succession studies.•Methodology based on wing measurements can be used to all Calyptratae flies of forensic importance. Correct species identification is a crucial step in research and the practice of forensic entomology. At the moment the most common and widely used tool for this purpose are taxonomic keys based on morphological characters. During recent decades molecular methods have been used more and more often for species identification. Automated species identification can also be based on wing measurements. This method proved to be reliable in the case of some groups of necrophagous Diptera from various geographical regions. Up to now, this method has not been applied to a wide range of species of European Calliphoridae. We have used 968 specimens representing 15 species and 6 genera. Identification was based on 15 landmarks located at wing vein junctions. Wing shape differed markedly between the studied species. The overall correct classification rate at the level of species was 97.1%. Seven species, including Calliphora vomitoria, Chrysomya albiceps, Phormia regina, Protophormia terraenovae and Lucilia sericata were identified without error. Lucilia caesar and Lucilia illustris, which are considered relatively difficult to distinguish, were correctly classified in 93.0% and 94.1% cases, respectively. The correct classification rate at the level of genera was 99.2%. The results presented here suggest that wing measurements are a promising alternative to other methods of identification of necrophagous Calliphoridae. This method can be particularly useful for identification of large samples of females collected during carrion succession studies. It can also be used to double check identification performed by a taxonomist using traditional methods.

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 black blow fly, Phormia regina (Meigen) (Diptera: Calliphoridae) is one of the most abundant carrion flies in North America. Calliphorids are important in agriculture and animal production, veterinary sciences, forensics and medical entomology. While the role of flies in the epidemiology of human and animal diseases is an active area of research, little is known about the microorganisms associated with these insects. We examined the diversity of wild-caught black blow fly endogenous (internal body) and exogenous (external body) microbial communities using 16S rRNA gene sequencing. Overall, 27 phyla, 171 families and 533 genera were detected, and diversity was significantly higher ( P < 0.05) on external body surfaces. At the genus level, Dysgonomonas , Ignatzschineria , Acinetobacter , Vagococcus , Myroides , and Wohlfahrtiimonas were predominant. Cloning and sequencing of nearly full-length fragments of the 16S rRNA gene showed that some of the species identified are known to be pathogenic to humans, animals, and plants. Myroides odoratimimus and Acinetobacter radioresistens are well-known, multi-drug resistant bacteria. These results provide a snapshot of the microbial communities harbored by adult black blow flies and call for more comprehensive studies to better characterize the role these flies may play in the transmission of pathogenic microorganisms.

The genetic structure of forensically important blow fly (Brauer & Bergenstamm) (Diptera: Calliphoridae) populations has remained elusive despite high relatedness within wild-caught samples. This research aimed to determine if the implementation of a high-resolution spatiotemporal sampling design would reveal latent genetic structure among blow fly populations and to elucidate any environmental impacts on observed patterns of genetic structure. Adult females of the black blow fly, Phormia regina (Meigen) (Diptera: Calliphoridae), were collected from 9 urban parks in Indiana, USA over 3 yr and genotyped at 6 polymorphic microsatellite loci. The data analysis involved 3 clustering methods: principal coordinate analysis (PCoA), discriminant analysis of principal components (DAPC), and STRUCTURE. While the PCoA did not uncover any discernible clustering patterns, the DAPC and STRUCTURE analyses yielded significant results, with 9 and 4 genetic clusters, respectively. Visualization of the STRUCTURE bar plot revealed N = 11 temporal demarcations indicating barriers to gene flow. An analysis of molecular variance of these STRUCTURE-inferred populations supported strong temporally driven genetic differentiation (FST = 0.048, F'ST = 0.664) relative to geographic differentiation (FST = 0.009, F'ST = 0.241). Integrated Nested Laplace Approximation and Boosted Regression Tree analyses revealed that collection timepoint and 4 main abiotic factors (temperature, humidity, precipitation, and wind speed) were associated with the genetic subdivisions observed for P. regina. A complex interplay between environmental conditions, the unique reproductive strategies of the blow fly, and the extensive dispersal abilities of these organisms likely drives the strong genetic structure of P. regina in the Midwestern US.

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.

Flies are frequently used for postmortem interval (PMI) estimations. These estimates are usually based on the age of larval or pupal specimens. However, the age defines only the minimum PMI. In order to move forensic entomology further, a method useful for the estimation of an interval preceding insect appearance on a corpse called the pre-appearance interval (PAI) is needed. Recently, it was demonstrated that the PAI of several carrion beetles is closely related to the temperature prevailing throughout this interval. Hence, it was postulated to estimate PAI from temperature. In order to check premises for using this approach with flies, a test of the relationship between adult or oviposition PAI and temperature was made for nine species of European flies. Data on PAI originated from pig carcasses decomposing under various temperatures. Adult PAI of Hydrotaea dentipes , Hydrotaea ignava , Hydrotaea similis , Phormia regina , and Stearibia nigriceps and oviposition PAI of S . nigriceps were exponentially related to temperature. Only S . nigriceps revealed a close relationship, demonstrating solid premises for PAI estimation from temperature alone. Adult and oviposition PAI of Calliphora vomitoria and adult PAI of Hydrotaea pilipes were not related to temperature. Adult and oviposition PAI of Lucilia sericata and Lucilia caesar responded similarly, with an abrupt and large increase in a narrow range of low temperatures and no response in a broad range of high temperatures. Probably, different mechanisms form the basis for the response of PAI to temperature in flies colonizing carcasses shortly after death and flies colonizing carcasses later in the decomposition process.

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.