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Background: Sarcophagidae along with Calliphoridae (superfamily Oestroidea) were known as important colonizers of cadavers and could be used to estimate the postmortem interval (PMI), through tracking the flies colonizing pattern and/or monitoring their growth rates on cadavers. Many previous researches discussed the impact of toxins in decomposing corpses, on the developmental stages of insects, which would affect the accuracy of PMI estimation. Clonazepam belongs to the benzodiazepines, and it is one of the most routinely used drugs to control humans' seizures. The current study aimed to investigate the effect of clonazepam on the developmental stages of Sarcophaga argyrostoma, one of the most widespread Sarcophagidae in Giza Governorate. Also, we investigate the ability of these developmental stages to detect the drug. Results: The current study used different concentrations of clonazepam (25, 50, and 100 mg/ml) as an in vitro application of the flesh fly S. argyrostoma. The drug has affected significantly the morphological measurements (weight, length, and width) of the different developmental stages, especially in the highest concentration (100 mg/ml). Quantitative and qualitative analyses were applied, by using high-performance liquid chromatography-mass spectrometry (HPLC–MS) and protein carbonyls amount respectively, to detect clonazepam in the second, early third, and late third larval instars and prepupa of S. argyrostoma. The results showed that the relationship between the concentration of the drug and its detection in the same developing instar is interdependent, using HPLC–MS. However, the drug faded from instar to the following one and transformed to its metabolite form. Measuring the protein carbonyls amount (OD/mg protein/min) revealed an elevation in the macromolecules damage, compared to the control groups, in almost all treated groups. Conclusions: The current data suggested that clonazepam has oxidative damage in S. argyrostoma. While HPLC–MS was efficient in measuring the concentration of the drug in the insect, protein carbonyls analysis was a time- and cost-saving method and could be used to detect the drug in insects qualitatively. [ABSTRACT FROM AUTHOR]

In forensic entomology, the presence of insects aids investigations by helping to understand the time since death. Insects are the first organisms that colonize and decompose the carrion. The main application of insect evidence is determining minimum post mortem interval (PMImin) either by analyzing insect developmental patterns or by estimating insect succession on the corpse. Age determination of insects has a pivotal role in the estimation of PMI. A range of techniques is available for estimating age, which provides accurate PMI in investigations. The present review critically discusses the applications of entomological evidence in PMI estimation and enlightens the factors influencing PMI calculations. Age estimation of insects is the primary task in PMI estimation. This review concentrates on methods used in age estimation, their strengths and weaknesses. Entomotoxicology analysis is a valuable tool in solving poisoning cases when dead bodies are recovered after a long period and this review also analyses the scope of entomotoxicology in the area of death investigations. [ABSTRACT FROM AUTHOR]

Carrion flies are trapped to study their succession pattern and record their diversity or establish laboratory colonies for various purposes. They are typically collected manually, which is not only challenging and time‐consuming, but may also result in incomplete or biased inventories. Passive trapping techniques commonly used damage the specimens and make them difficult to identify.We describe the construction of a trap to capture sarcosaprophagous Diptera alive. We tested six traps using six rabbit carcasses (Oryctolagus cuniculus L.) per trial over 1 year and recorded the diversity and succession of insects. We also established fly colonies of Lucilia sericata (Meigen) adults using captured individuals.A total of 27,428 insects (i.e., 8669 specimens in spring, 7969 specimens in summer, 8417 specimens in autumn and 2373 specimens in winter) belonging to 21 families and 43 species were collected over a period of 23, 19, 24 and 30 days in spring, summer, autumn and winter, respectively. The catch consisted mainly of the order Diptera (98.55%), but also Hymenoptera (1.45%). We successfully started a fly colony with 107 individuals in spring and added wild‐type adults every season. The colonies were maintained for 1 year, during which the lifespan of the adult flies varied between 5 and 52 days.The device is easy to manufacture, low‐cost, robust under field conditions, efficient and time‐saving. It enables large samples to be collected within a short period of time over the entire trial period without affecting its condition or performance. [ABSTRACT FROM AUTHOR]

A year-long entomological survey was performed in 2016-2017 using carrion-baited traps (chicken, pork, squid, or beef) to assess the diversity of necrophagous flies in the municipality of Yautepec, Morelos, Mexico. A total of 4,765 flies from 18 families were collected, more than 50 percent of the specimens collected (2,644) belonged to Calliphoridae represented by five genera and eight species. Out of these eight species we evaluated if the species presented a preference for a specific type of bait, our results show that Chrysomya rufifacies and C. megacephala preferred squid, while Cochliomyia macellaria, Hemilucilia segmentaria, and Lucilia mexicana preferred chicken. [ABSTRACT FROM AUTHOR]

Further to a previous publication by the European Council of Legal Medicine (ECLM) concerning on-site forensic and medico-legal scene and corpse investigation, this publication provides guidance for forensic medical specialists, pathologists and, where present, coroners' activity at a scene of death inspection and to harmonize the procedures for a correct search, detection, collection, sampling and storage of all elements which may be useful as evidence, and ensure documentation of all these steps. This ECLM's inspection form provides a checklist to be used on-site for the investigation of a corpse present at a crime or suspicious death scene. It permits the collection of all relevant data not only for the pathologist, but also for forensic anthropologists, odontologists, geneticists, entomologists and toxicologists, thus supporting a collaborative work approach. Detailed instructions for the completion of forms are provided. [ABSTRACT FROM AUTHOR]

Given the growing demands for standardization of procedures for collection, preservation, and identification of insect specimens, this study aimed to identify how forensic entomology is used in Brazil and the perceptions of professionals involved in this process. We conducted: (1) bibliometric survey of the scientific production of forensic specialists from 2001 to 2020; (2) questionnaire applied to forensic professionals to verify their perceptions about the use of entomological evidence in practice. An increase in publications on Forensic Entomology was noticed in the last 5 years, mainly in the South and Northeast regions. Of the 82 respondents to the questionnaire, 62.8% deal with cadavers at least once a week, but 89.0% of the professionals who examine cadavers weekly find less than 10.0% colonized by insect larvae. Using entomological evidence to estimate PMI was mentioned by only 36.6% of the professionals. Most respondents stated no procedures related to collecting, handling, or identifying necrophagous insects in their daily practice. Regarding the chain of custody, 64.6% said they were aware of the changes in the regulations involving the collection and maintenance of forensic evidence. We emphasize the importance of collaboration between scientists at universities and crime scene examiners. Although the use of entomological evidence in criminal investigations in Brazil is still scarce, a growing interest in this area can be detected among forensic experts. [ABSTRACT FROM AUTHOR]

The feeding of vertebrate scavengers on animal remains has been noted for at least 30 years in relation to the creation of postmortem artifacts. However, the subsequent impact on other necrophagous arthropods, which interact with these remains has not been well documented. Herein, we report a rodent (Rattus spp.) feeding event that altered a perimortem wound beneath the jaw of a decomposing swine carcass. Point trauma such as this has been cited as insufficient for insect colonization; however, the resulting enlargement of the wound due to scavenger feeding has the potential to serve as an oviposition site where colonization would typically not be expected otherwise (i.e., in the absence of a wound or the presence of a small wound such as point trauma). In fact, colonization was observed surrounding the site of scavenger alteration. If scavenger artifacts (e.g., postmortem alteration of remains by feeding) are not identified appropriately and recognized for the effects they may have on necrophagous arthropods then associated assessments in forensic investigations could be affected. [ABSTRACT FROM AUTHOR]

Forensic entomology aids investigations using insects and is primarily associated with the estimation of post‐mortem interval (PMI). Studies have shown that human DNA can be recovered from the crops of fly larvae. While several factors regarding the recovery of human DNA from crops have been studied, DNA extraction methods have not been thoroughly assessed. Determining a method for optimal extraction could aid crime laboratories in implementing DNA extraction from larvae and streamlining future research. Bovine DNA was used as a substitute for human DNA to test several DNA extractions kits. Four DNA extraction kits (Chelex®, PDQeX forensicGEM, EZ1® DNA Investigator, DNeasy® Powersoil® Pro Kit) were evaluated based on the quantity and quality of bovine DNA extracted. Extractions were performed on whole fly larvae and dissected crops. Quantification was performed using real‐time PCR (qPCR) on a StepOne™ Real‐Time PCR System with SYBR® Green using bovine‐specific cytochrome b primers. The quality of extracts was determined by checking for inhibition using commercial qPCR chemistries with an internal PCR control (IPC). When using whole fly larvae, Powersoil® Pro yielded the highest average DNA yield (n = 10, 0.668 ± 0.458 ng/μl), while EZ1® DNA Investigator yielded the highest average with crops (n = 10, 0.605 ± 0.403 ng/μl). Chelex and forensicGEM yielded low amounts of bovine DNA, and its extracts were inhibited, unlike EZ1® and Powersoil® Pro, which have purification steps. Therefore, it is recommended to use EZ1® DNA Investigator coupled with automation on EZ1® Advanced XL to recover DNA from fly larvae crops. [ABSTRACT FROM AUTHOR]

Simple Summary: Forensic entomology applies the knowledge of arthropods to obtain useful information for the resolution of forensic investigations. In legal medicine, one of the methods used for the dating of death is the faunal succession, which is based on the orderly and predictable changes of the species associated with a corpse over time. The potential of insects for this purpose has been known for more than seven centuries, but mites are only currently being incorporated. Mites can provide useful information on the time and place of death because: they are a very diverse group, ubiquitous, abundant, and frequent; they contribute to the faunal succession; they are indicators of location and transfer of the corpse; they can be present in unfavorable conditions or environments for insects; they complement the information obtained from insects; and they are tracking evidence. Due to the absence of knowledge regarding the sarcosaprophagous fauna in the Mediterranean region, it is necessary to highlight the diversity of insects, together with their phoretic mites, present in decomposing animal organic matter, and their interaction network in this region, in order that the results obtained can be applied in future forensic investigations and contribute to the estimation of the time and place of death. The potential of insects for forensic investigations has been known for more than 700 years. However, arthropods such as mites could also play a role in these investigations. The information obtained from insects, together with their phoretic mites, is of special interest in terms of estimating the time and geographical location of death. This paper presents the first interaction network between phoretic mites and their host insects in Navarra. It also reports the first time that an interaction network was applied to animal remains of forensic relevance. The data reveal the degrees of specificity of the interactions established, the biological and ecological characteristics of the mites at the time of association, and factors that played important roles in the mites' dispersion. Fauna was collected using 657 traps baited with 20 g of pig carrion over a year. Only 0.6% of insects collected carried phoretic mites. The network comprised 312 insects (275 beetles, 37 flies) and 1533 mites and was analyzed using various packages of the R programming language. We contribute new host insect records for 15 mites, 3 new records of insects as hosts, 5 new mite records for the Iberian Peninsula, and 2 new mites records and 8 new insect records for Navarra. [ABSTRACT FROM AUTHOR]

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Abstract Background Studies on the insect fauna of animal corpses, used as a vertebrate model, may help forensic investigation cases to estimate the post-mortem-interval (PMI), cause of death, and crime-scene location. Likewise, entomofauna of buried corpses can assist in determining the post-burial-interval (PBI), movement of the body or hiding of the crime. The bodies buried under the soil decompose at a slower rate than the body exposed. Also, there are fewer insects that can go underground to locate the corpse. Such types of studies on the insect fauna of buried carrion can help in forensic investigation cases. The current study aims to determine the succession of a goat carcass buried shallowly in an outdoor habitat of the Aligarh Region, North India. Results In the present study, we examined a goat carcass buried at a depth of nine inches to determine the type of insects capable of colonizing buried animal carcass in the study area of Aligarh, Uttar Pradesh, India. We have found five species of Diptera, three species of Coleoptera, one Hemipteran species, and one Hymenopteran species. Adult Dipterans found during different samplings were Megaselia scalaris (Loew 1866), Chrysomya megacephala (Fabricius 1794), and Calliphora vicina (Robineau-Desvoidy, 1830). Dipterans’ immature stages were found to be of Synthesiomyia nudiseta (van der Wulp, 1883) and Hydrotaea capensis (Wiedemann, 1830) Megaselia scalaris. Adults of Saprinus quadriguttatus (Fabricius, 1798), Saprinus splendens (Paykull, 1811), and Onthophagous quadridentatus (Fabricius, 1798) of order Coleoptera. Adult Cydnus species (Fabricius, 1803) of Hemiptera and Dorylus species (Fabricius, 1793) of Hymenoptera have also been recorded. During the sampling, the authors reported the stages of carcass disintegration and the insects associated with it. Conclusions Flies, beetles, and ants have been recorded in association with the buried goat carcass, which may add to the knowledge of colonization of buried bodies in India and around the globe. S. nudiseta, H. capensis, and M. scalaris, which are found on the goat carrion in the current study, have also been reported on human corpses in the past. Furthermore, M. scalaris found consistently on two sampling dates beneath the ground on the goat carcass and was also seen running on the grave’s surface. In addition, authors have reported several species belonging to different order and families, corresponding to various stages of decomposition of corpse which were earlier not known from buried corpses from India. So, it is crucial and may have implications in medicolegal cases.

The objective of this study was to identify insects species visiting decomposing rabbit carrion and to establish the relationship between insects and decomposing carrion which may be of forensic importance in Jos, Plateau state Nigeria. Six rabbit weighing between 2.3kg and 2.7kg were killed by a sharp blow with a blunt metal object on the heads. Two each of the six rabbit were exposed to the sun, under shade and near river. The carcasses were allowed to decompose and insect were collected twice a day for the first one week and thereafter once a day up to the end of the 15 weeks. Four decomposition stages recorded were fresh, bloating, decay and post decay stages. A total of 16 insect species from 7 families and 3 orders were observed. Three Diptera families (Calliphoridae, Muscidae and Sarcophagidae) were identified from carcasses in all the locations and they were the first to be observed. Species collected included Lucilia Cuprina, Chrymosya albiceps, Chrymosya megacephala, Phormiaregina, Musca domestica and Hydrotaea sp. Three Coleopteran families (Cleridae, Silphidae and Dermestidae) were identified from all the carcasses and the observed species were Necrobia rufipes, Necrophile rufithorax and Dermestes muculatus. Hymenoptera (Formicidae) was also present throughout the period. The study shows that insects from the order Diptera (Calliphoridae and Muscidae) indicate the fresh and bloat stage, while the order Coleopteran (Dermestidae) indicate the decay and post decay stage during period of decomposition of the carcasses and could be of importance for further forensic studies since they were collected from the carcasses.

Abstract Background Evaluation of insect succession patterns is a scientific method to estimate the time elapsed since death. Several studies have shown that ante-mortem intake of opioids affect maggot growth rate. However, there are few published data that investigate the effect of ante-mortem opioid use on insect succession patterns. Therefore, the main purpose of this research was to investigate the effect of methadone on the succession patterns of insects on rabbit carcasses during the spring and winter of 2019. In the present study, the H-null hypothesis represents the dissimilarity between the successional waves of species from testing and control carcasses (H0: P: 0). Results During this study, 15 and 13 insect species were collected from carcasses during the spring and winter, respectively. The most dominant species during the both seasons were Chrysomya albiceps and Calliphora vicina. These two species preferred to lay eggs on the control carcasses earlier than the treated carcasses. Lucilia cuprina was observed only from the remains of untreated carcasses, while Saprinus chalcites and Necrobia rufipes were recorded only from the remains of treated rabbits. Samples indicate that 11.8% of the insects were members of the Coleoptera. Permutation analyzes based on the Mantel test were 0.647 ± 0.16 (P = 0.009) and 0.693 ± 0.16 (P = 0.003) for the similarity of the species between treated and untreated carcasses in the spring and winter, respectively. Permutation analyzes for the two most dominant fly species (Ch. albiceps and C. vicina) between the treated and untreated carcasses in the spring and winter were 0.515 ± 0.15 (P = 0.05) and 0.491 ± 0.14 (P = 0.09), respectively. Conclusion The results revealed that the overall pattern of insect succession was similar between the treated and untreated rabbit carcasses. However, the patterns of succession of Chrysomya albiceps and Calliphora vicina differed slightly between treated and untreated carcasses, and this could have an effect on the PMI min estimate.

Global climate change and, specifically, rising temperatures, may increase the number of generations of necrophagous insects. The common green bottle fly Lucilia sericata (Meigen, 1826) (Diptera, Calliphoridae) ranks among the most important cosmopolitan necrophagous insects that utilize corpses and cause myiasis in farm animals and humans. Based on the data simulations, the use of accumulated degree-hours enables to calculate the number of generations of this forensically important species of blowfly with a greater accuracy than before, considering short-term increases of temperature at the boundary of the cold and warm seasons. The number of generations of L. sericata has increased from 7.65 to 8.46 in the Ukrainian steppe zone over the last 15 years, while the active developmental period of this species has increased by 25 days due to earlier start in spring. The average temperature increase of 1 °C increased the number of generations of L. sericata by 0.85. With a global climate change following the Representative Concentration Pathway (RCP) 4.5 scenario (average temperature increase of 2.4 °C), adopted by the Intergovernmental Panel on Climate Change, by 2100 the number of generations of L. sericata in a simulated ecosystem will increase by 2.0 to 9.0 generations per year.

[Display omitted] • We studied Phormia regina pupae age estimation using ATR-FTIR and machine learning. • The concentration of amide I band in puparium gradually decreases over time. • Pupal age can be estimated with 88% accuracy, surpassing morphological constraints. • Compared to random forest, the PLS model is more suitable for estimating pupal age. • There are latent age indicators in the ATR-FTIR spectral data. Accurate postmortem interval estimation is vital in the investigation of homicides, suicides, and accidental deaths. It is key in narrowing suspect lists, improving crime-solving efficiency, and offering solace to bereaved families. The intra-puparial period, comprising about half of a fly's developmental cycle, presents challenges for morphological age estimation. External changes are limited to color shifts and the appearance of respiratory horns on the puparium only within several hours after pupariation, while detailed internal development analysis often requires invasive methods like removing the puparium, which can be damaging. Additionally, these techniques usually depend on a forensic entomologist's expertise, which lead to subjective biases. This study employed attenuated total reflection-fourier transform infrared spectroscopy, a rapid, non-destructive method for analyzing proteins, chitosan, and chitin in puparia. Data showed a consistent reduction in the concentration of the amide I band within the puparium during the intra-puparial development at five constant temperatures (19 °C, 22 °C, 25 °C, 28 °C and 31 °C). This trend in the spectral data effectively distinguishes pupae at various stages of intra-puparial development, facilitating precise age estimation, which is critical for the estimation of the minimum postmortem interval (PMI min). Finally, this work combined the total reflection-fourier transform infrared spectroscopy with chemometric analysis and successfully developed a partial least squares discriminant analysis model and a random forest model, with accuracies of 88 % and 81 %, respectively. These models enable the non-invasive age estimation of P. regina in its intra-puparial period, a stage traditionally difficult to assess morphologically, thus laying the groundwork for PMI min estimation using fly pupae. [ABSTRACT FROM AUTHOR]

Forensic entomology is primarily concerned with the estimation of time since death and involves determination of the age of immature insects colonising decomposing remains. Accurate age determination of puparia is usually accomplished by dissection, which means destructive sampling of evidence. As part of improving abilities to correctly identify species and developmental age, it is highly desirable to have available non-destructive methods. In this study, we acquired external hyperspectral imaging (HSI) data (77 spectral bands, 389-892 nm) from the dorsal and ventral sides of individual puparia of two species of blowfly (Diptera: Calliphoridae), Calliphora dubia Macquart 1855 and Chrysomya rufifacies Macquart 1842. Puparia were dissected to determine the presence/absence of eight internal morphological development characteristics (legs, wings, labella, abdominal segments, antennae, thoracic bristles, orbital/facial bristles and eye colour and arista). Based on linear discriminant analysis and independent validation of HSI data, reflectance features from puparia could be used to successfully (1) distinguish the two species (classification accuracy = 92.5 %), (2) differentiate dorsal and ventral sides of puparia (classification accuracy C. dubia = 81.5 %; Ch. rufifacies = 89.2 %) and (3) predict the presence of these morphological characteristics and therefore the developmental stage of puparia (average classification accuracy using dorsal imaging: C. dubia = 90.3 %; Ch. rufifacies = 94.0 %). The analytical approach presented here provides proof of concept for a direct puparial age relationship (i.e. days since the onset of pupation) between external puparial reflectance features and internal morphological development. Furthermore, this approach establishes the potential for further refinement by using a non-invasive technique to determine the age and developmental stage of blowflies of forensic importance.

Objective To identify species of common sarcosaprophagous flies based on digital image analysis of veins, in order to provide new idea for fast and accurate species identification of sarcosaprophagous flies in forensic entomology. Methods Random trapping of 226 male and female sarcosaprophagous flies that comprised of 7 common species, including Sarcophaga peregrina, Parasarcophaga ruficornis, Sarcophaga dux, Seniorwhitea reciproca, Bercaea cruentata, Aldrichina grahami, and Synthesiomysia nudiseta with carrion in the field was conducted. The 17 landmarks on the right wing of each fly were digitally processed and the images were analyzed. The effects of allometry were evaluated using a permutation test. Wing shape variations among 7 sarcosaprophagous fly species and female species was analyzed using canonical variate analysis （CVA）. Additionally, cross-validation test was used to evaluate the reliability of classification. Results Among 7 sarcosaprophagous fly species and female species, the effect of allometry had statistical significance （P

Estimation of postmortem interval （PMI） has always been one of the difficult problems for forensic scientists. It is especially hard to estimate the PMI of highly decomposed corpses in the wild or in secluded houses with conventional methods. Therefore, application of insect evidence at the scene is usually required for estimation. Sarcosaprophagous flies of different species have totally different developmental rates. In actual cases, direct measurement of the body length of the larvae, calculation of accumulated temperature and succession stages without species identification, or calculation based on incorrect species identification would often lead to a large deviation between the calculated results and the real PMI. This mistake would also mislead the case investigation. Therefore, accurate species identification should be implemented before any PMI estimation of decomposed corpses with forensic entomological methods. This article reviews the general and ultramicroscopic species identification and molecular biological species identification methods of different stages of sarcosaprophagous flies, in order to provide new ideas and methods for related research and practice, and provide reference for the application and promotion of forensic entomology in the front line of public security.

Entomological evidence provides entry points and clues for cases detection, in terms of estimation of the postmortem interval （PMI）, and place and cause of death. In recent years, the feasibility of entomological evidence in practice has been proved by theories and cases. It especially plays an important role in the investigation of cases with unnatural death, no monitoring, and highly corrupt cadaver. However, there are still some key issues to be further studied and standardized before the application of entomological evidence to forensic practice, to improve the effect of entomological evidence in forensic investigation and trial. This paper retrospectively reviews key studies of the application of entomological evidence in forensic science, mainly including discussion of forensic entomology inspection standard, identification studies of sarcosaprophagous insect species, collection of sarcosaprophagous insect growth and succession data under different environments and forensic entomotoxicology. With the rapid development of information technology and biotechnology, applying artificial intelligence and whole genome sequencing technology in forensic entomology has become a new research direction, which can improve the application value and range of entomological evidence in forensic science.

Objective To test the feasibility and accuracy of with sarcosaprophagous insects postmortem interval （PMI） estimation with sarcosaprophagous insects and provide references for estimation practice. Methods Eleven cases confirmed by the detection results, with complete entomological evidence were selected. The insect species, estimation results and true results involved in the cases were statistically analyzed and compared. Results Thirteen species of insects were found at the criminal scene, including Chrysomya megacephala （Fabricius）, Chrysomya rufifacies （Macquart）, Chrysomya nigripes （Aubertin）, Lucilia sericata （Meigen）, Hydrotaea spinigera Stein, Muscina stabulans （Fallén）, Sarcophagid （species were not identified）, Megaselia scalaris （Loew）, Hermetia illucens （Linnaeus）, Saprinus splendens （Paykull）, Creophilus maxillosus （Linnaeus）, Dermestes maculatus （De Geer） and Necrobia ruficollis （Fabricius）. The PMI of all eleven cases was within the range of estimated PMI. The estimated results of 72.73% cases were on the same day of the true results. Conclusion Sarcosaprophagous insects can estimate the PMI simply and conveniently. In cases where the PMI is within the time range of one generation of flies or beetles, the estimation results are relatively accurate. However, the estimation is less accurate when the PMI is beyond the time range.

Insect samples found on human corpses can provide the information important to estimating the minimum postmortem interval （PMImin）. A female cadaver, found in a deserted factory in Chongqing of China, was confirmed as a homicide case after the forensic investigation and autopsy. Determining the time of death was difficult due to the inconsistent degree of decomposition in different parts of the decedent. The insect specimens found on the cadaver were identified to be Chrysomya rufifacies （C. rufifacies, Macquart） by morphology and mitochondrial DNA sequence analysis. The PMImin was estimated to be 452 h, based on the developmental rate of C. rufifacies. The PMImin was estimated successfully to be almost precise, which provided an important entomological evidence for case investigation and suspect prosecution. In so doing, this highlights the usefulness of entomological evidence of specific species in the geographic area for PMI accurate estimation, especially in the case of advanced decomposed corpses.

Objective To establish the basic data for estimating minimum postmortem interval （PMImin） of heavily decayed and skeletonized remains by studying the development of Dermestes maculatus DeGeer （Coleoptera: Dermestidae）. Methods The developmental stages of Dermestes maculatus were observed at four constant temperatures of 20 ℃, 24 ℃, 28 ℃ and 32 ℃, and the changes in body length were also examined as the biological indicator to estimate larval day-age and instar. Results The total developmental time from egg to adult at 20 ℃, 24 ℃, 28 ℃ and 32 ℃ were （126.7±10.6） d, （69.4±8.2） d, （50.4±8.4） d and （49.6±6.5） d, respectively. The body length increased gradually, but changed irregularly as a whole. Conclusion The study provides basic data on the development and growth of Dermestes maculatus, especially on its developmental duration as a significant value for estimating PMImin of heavily decayed and skeletonized remains. Nevertheless, the change of body length is not found to be the best biological indicator for instar determination.

Blow flies (Calliphoridae) are used in forensic entomology studies, because their life cycle help to determine the minimum post-mortem interval (PMImin). Paralucilia fulvinota (Bigot, 1877) is a forensic indicator and is reported from human corpses in the Amazonian region. Our objectives were to describe the development time of P. fulvinota under natural conditions at Ducke Reserve (Amazonas), provide the accumulated degree-days (ADD) and accumulated degree-hour (ADH), body length and mass of larvae, and the sex ratio of adults. Pregnant females were sampled on a swine carcass. From hatched eggs, we reared stock colonies and observed the immature development under uncontrolled conditions of temperature (24.5 ± 1.3 ºC), rainfall (188 ± 11.5 mm³) and humidity (84 ± 3.9%). Development time was 11 days. Adult sex ratio was 0.86. Larvae gained mass at constant rates through the instars but grew less in length from the second to the third instar. Accordingly, P. fulvinota grows more in mass than in length when in larval stage. The period from first stadium to adults lasted 159.5 degree-days and 3828 degree-hours. Our results may be helpful in providing a more accurate estimate of PMImin on corpses encountered in forested areas of the Amazon region.

Flies (Diptera) have played a prominent role in human history, and several fly species are reared at different scales and for different beneficial purposes worldwide. Here, we review the historical importance of fly rearing as a foundation for insect rearing science and technology and synthesize information on the uses and rearing diets of more than 50 fly species in the families Asilidae, Calliphoridae, Coelopidae, Drosophilidae, Ephydridae, Muscidae, Sarcophagidae, Stratiomyidae, Syrphidae, Tachinidae, Tephritidae, and Tipulidae. We report more than 10 uses and applications of reared flies to the well-being and progress of humanity. We focus on the fields of animal feed and human food products, pest control and pollination services, medical wound therapy treatments, criminal investigations, and on the development of several branches of biology using flies as model organisms. We highlight the relevance of laboratory-reared Drosophila melanogaster Meigen as a vehicle of great scientific discoveries that have shaped our understanding of many biological systems, including the genetic basis of heredity and of terrible diseases such as cancer. We point out key areas of fly-rearing research such as nutrition, physiology, anatomy/morphology, genetics, genetic pest management, cryopreservation, and ecology. We conclude that fly rearing is an activity with great benefits for human well-being and should be promoted for future advancement in diverse and innovative methods of improving existing and emerging problems to humanity.

Forensic entomology plays a crucial role in death investigations, particularly in estimating the postmortem interval (PMI). This study presents a forensic entomology case involving a corpse found in a buried utility vehicle. The victim was in an advanced state of decomposition, with autopsy findings revealing gunshot wounds. Cadaveric fauna was collected at the scene and during the autopsy. The analysis revealed a diverse insect community, with predominance of Compsomyiops fulvicrura and Piophila casei. The time of development of species like Dermestes maculatus and Necrobia rufipes was used to estimate the minimum PMI. The presence and low abundance of Calliphora vicina, a species preferring lower temperatures, shed light on the seasonal conditions at the time of death and suggested possible body concealment shortly after death. This research is the first to report insects as evidence in a corpse found in a buried vehicle and contributes to the body of knowledge in forensic entomology. The study also suggests that the use of entomological evidence can provide additional information about the season in which the body was concealed, making it a valuable tool in death investigation and crime scene reconstruction. Finally, it emphasizes the need for proper sampling, expert identification, and close collaboration between forensic entomologists and pathologists. [ABSTRACT FROM AUTHOR]

Blow flies (Diptera: Calliphoridae) are frequently used in forensic investigations due to their rapid colonization of cadavers. As with other insects, environmental temperature strongly influences their developmental rates. While published research has typically explored not only the impact of the environmental temperature, but also of other factors like tissue type and drug presence on developmental rates, the influence of photoperiod on the developmental rates of forensically relevant blow fly species has remained largely underexplored. Understanding the relationship between photoperiod and developmental times is crucial, as neglecting this aspect could compromise the accuracy of minimum post-mortem interval (minPMI) estimations. The present study investigates the impact of three photoperiod conditions (0:24, 8:16, and 12:12 light:darkness) on the developmental rates of Calliphora vicina , focusing on the duration of the different immature stages and on the total developmental time. Our results revealed significant variation in the intra-puparial stage and total development time across different photoperiods. Notably, a 12:12 photoperiod led to a significantly prolonged intra-puparial stage and total development time compared to the 0:24 photoperiod, suggesting that Calliphora vicina develops faster in total darkness. These findings highlight the importance of considering photoperiod in both laboratory rearing protocols and forensic casework to improve the accuracy and reliability of minPMI estimations. In this regard, preliminary guidelines and recommendations are provided. • The photoperiod regime significantly influences the duration of blow fly intra-puparial period. • Consequently, the total developmental time is significantly affected by photoperiod. • Preliminary recommendations on photoperiod for developmental studies are provided. [ABSTRACT FROM AUTHOR]

Only a few cases describing the activity of dermestids on human corpses have been reported in the literature. Dermestes maculatus is a cosmopolitan beetle associated with carcasses at different decomposition stages, usually colonizing skeletonized and mummified remains. In this study, we presented two forensic case reports of D. maculatus associated with human corpses in Santa Catarina State, southern Brazil. In the first case, a human corpse in an advanced stage of decomposition was found hanged in an outdoor urban area. In the other one, a mummified cadaver was found inside of a closed house. In this last case, larvae of D. maculatus were essential to estimate the minimum postmortem interval (PMI) for the first time in Scientific Police of Santa Catarina. Our records highlight the significance of necrophagous beetles in ecological succession and in estimating PMI in cases involving human remains and corpses in advanced stage of decomposition. • Necrophagous beetles can be useful to estimate the postmortem interval (PMI). • L7 larvae of Dermestes maculatus were used to estimate PMI of a mummified corpse. • The minimum PMI was estimated in 44 days. [ABSTRACT FROM AUTHOR]

There is a significant gap in the availability of comprehensive identification keys for the early larval stages of forensically important fly species. While well-documented identification keys exist for the third instar larvae, particularly for the Calliphoridae, Muscidae and Sarcophagidae families, there is a notable scarcity of keys for the first, except Calliphoridae, and the second instar larvae, with no such resources available for muscid species. The second instar larvae suffer the most from the lack of morphological descriptions and available identification keys. The Muscidae is one of the most frequently reported dipteran families of forensic importance colonising animal cadavers and human corpses. Nevertheless, descriptions of the morphology of their early instars remain scarce and limited to only a few species, thus their larval identification is challenging or impossible. Considering the numerous challenges associated with studying small-sized entomological material, we tested whether it is feasible to identify muscid flies to the species or at least genus level based predominantly on the details of the cephaloskeleton. To overcome the obstacle of observing details of small sclerites, especially their shapes and interconnections, we effectively employed confocal laser scanning microscopy (CLSM) as a supplementary method for light microscopy (LM). This study provides an identification key for first and second instar larvae of forensically important muscid species from the western Palaearctic (Europe, North Africa, Middle East). The proposed key primarily utilises details of the cephaloskeleton with only addition of external morphology. • The cephaloskeleton of early instars of 14 necrophagous muscid species was examined using light and confocal microscopy. • Detailed examination of the cephaloskeleton enabled differentiation of muscid larvae to the species-level. • The first identification key for the early instars of Muscidae is provided. • Detailed morphological documentation of early instars muscid larvae is provided. [ABSTRACT FROM AUTHOR]

Analysis of insects can provide evidence in death cases, for example, by answering the question about the time of death. Apart from flies, beetles are the second most useful insect group in forensic entomology. To elucidate the time of death based on insect evidence, developmental models of a given species are necessary. In this study, we developed such models for Necrodes littoralis, a necrophagous beetle, which is common in the Palearctic region and has great potential in forensic entomology. We monitored the development at 10 constant temperatures (14–30 °C). Larvae were reared in aggregations. Thermal summation models, isomorphen and isomegalen diagrams and growth curves were derived using the data. Depending on the temperature, development lasted between about 23 and 89 days. Mortality was high at the extremes of the temperature range. The thermal summation constant for the total development was 434.7 ± 28.86 accumulated degree-days above a developmental threshold of 9.04 ± 0.55 °C. This is the first comprehensive dataset on the development of N. littoralis. Implications for its use in forensic casework are discussed. [ABSTRACT FROM AUTHOR]

Simple Summary: From the point of view of forensic entomology, the study of the development times of the species and the factors conditioning them are issues of great importance to estimate the minPMI. Hydrotaea capensis is a Muscidae (Diptera) of forensic interest because of its colonization preferences. Thus, there is a need to have more precise data about its biology to gain insights into the interpretation of its presence in corpses. In this study, the H. capensis life cycle was studied at four constant temperatures, 18, 20, 25 and 30 °C, by recording the duration of its different developmental stages and the length reached in each larval stage, as well as some biometric characteristics of the emerged adults. Our work is the first to provide data on the duration of the life cycle of this species and pointed to some alar features as relevant biometric adult indicators to be considered. One of the most important and perhaps most used applications of forensic entomology concerns the estimation of the minimum post-mortem interval (minPMI), defined as the time interval between death and the actual finding of a corpse. Some Diptera species are critical in these studies because they are the first ones capable of detecting and finding a corpse and are selectively attracted by its decomposing status. Thus, the knowledge of the micromorphology of their preimaginal stages and of their life cycles within a time frame constitutes solid indicators for estimating the minPMI. Hydrotaea capensis is a Muscidae of forensic interest usually considered as a late colonizer of corpses. It is widely distributed, living mainly in warm regions, and present in a wide variety of habitats. In this study, the H. capensis life cycle was studied at four constant temperatures, 18°, 20°, 25° and 30 °C, by recording the duration of its different developmental stages, including the length reached in each larval stage, as well as some biometric characteristics of the emerged adults. Significant differences were observed in the average time of development of most larval stages, with a longer duration at low temperatures, and in the length of each larval stage depending on the temperature, but, in this case, without a clear pattern. Moreover, significant differences were found in some alar features, pointing to them as a relevant indicator to be considered. The data provided will assist forensic entomologists to make more accurate minPMI estimations in cases where H. capensis is present. [ABSTRACT FROM AUTHOR]

This study explored the trade‐offs between thermal regulation and aggregation, two key factors impacting blow fly (Diptera: Calliphoridae) larvae development.Recent works have demonstrated that necrophagous maggots engage in developmental niche construction, which provides adaptive benefits. First, each species has a preferential temperature, at which larvae grow fast and efficiently. Second, larvae are attracted by each other and aggregate in large maggot masses. These groups modify the local environment and facilitate the exodigestion process (niche construction by perturbation). However, aggregation and relocation towards thermal preferendum are not always compatible under field conditions, forcing larvae to make choices.To test the developmental consequences of such trade‐offs, 40 or 80 Lucilia sericata larvae were placed on a thermal gradient (from 22 to 48°C) with or without a captive aggregate of 40 larvae located at 22°C, and their development speed, size and survival were measured.A previous study showed that in such situation the free larvae alone relocated at 33°C, while in the presence of captive larvae they gathered with the captive group at 22°C. In the present developmental study, we observed that such 22°C aggregated larvae actually grew as fast as if they were at 33°C.This result shows that niche construction, here resulting from larval gregarism and feeding facilitation, can compensate for the physiological costs of low ambient temperature. This finding confirms that aggregation of necrophagous Diptera larvae is an efficient adaptation to the carrion environmental constraints and highlights the adaptive value of developmental niche construction. [ABSTRACT FROM AUTHOR]

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Assessing the time of death based on the growth and development of insects is a critical task in forensic entomology. The rate of larvae development can be affected by a variety of toxins, including pesticides. Aluminum phosphide (AlP) is a low-cost insecticide that has yet to be tested for entomotoxicological significance, despite the fact that it is frequently the cause of fatal poisoning. In this study, we measured the body length of Chrysomya albiceps larvae reared on the carcasses of rabbits poisoned with AlP and analyzed the morphological changes of the larvae reared on the carcasses of rabbits poisoned with AlP. The concentration of AlP in the body of the larvae was significantly lower than in rabbit tissues. Insects from the AlP group had a significantly lower gain in body length. Furthermore, deformities in the larvae were found. Smaller respiratory spiracles were found, as well as a deformed small posterior end with hypogenesis of the posterior respiratory spiracles. Thus, disturbed growth and development of carrion flies found at a crime scene could indicate pesticide poisoning, such as aluminum phosphide. [ABSTRACT FROM AUTHOR]

Necrophagous blowflies (Diptera: Calliphoridae) are of great importance particularly during investigations of suspicious deaths. Many studies have analyzed the distribution of blowflies based on pig experiments and baited trapping; however, data from real case scenarios are rarely used. In this article, the distribution of blowflies found during investigations of 160 real cases during 1993–2007 in Switzerland is evaluated based on habitat, altitude, and season. Ten species of blowflies were present in 145 out of the 160 cases. The most common species was Calliphora vicina, which occurs throughout the year and was present in 69 % of all cases. Lucilia sericata, Calliphora vomitoria, and L. caesar were identified among the rest of the flies as species of great forensic importance mainly due to their distributional patterns. After a comparison with a similar dataset from Frankfurt, Germany, some surprising differences were determined and discussed. The biggest discrepancies between our dataset and the German dataset were in the occurrences of L. sericata (30 % vs. 86 %, respectively), Phormia regina (5 % vs. 43 %), and L. ampullacea (1 % vs. 45 %). The life-history strategies and intraspecific behavioral variability of blowflies remain understudied, although they can be essential for an unbiased approach during a death investigation. Further research and comparison of occurrence patterns across the area of distribution of blowflies are therefore needed and recommended. [ABSTRACT FROM AUTHOR]

Simple Summary: Forensic entomology plays a major role in postmortem interval (PMI) estimation, mainly based on morphological characteristics of larvae colonized on decomposed corpses. However, weight and length cannot be used as relevant parameters to calculate the age of wandering at the larval and pupal stages. In order to identify a specific indicator that may serve as a potential target, studies on the mechanism during postembryonic development may provide a foundation to establish a molecular age determination method. Sarcophaga peregrina (Robineau-Desvoidy, 1830) is a species of medical and forensic importance. In this study, transcriptome analysis was performed to investigate gene expression profiles from the larval to pupal stages of S. peregrina, which were used to identify differentially expressed genetic markers as candidates for molecular age estimation. This study explores molecular mechanisms during postembryonic development of S. peregrina. The developmental, stage-specific gene profiles suggest genetic markers for age prediction of forensically important flies. Sarcophaga peregrina (Robineau-Desvoidy, 1830) is a species of medical and forensic importance. In order to investigate the molecular mechanism during postembryonic development and identify specific genes that may serve as potential targets, transcriptome analysis was used to investigate its gene expression dynamics from the larval to pupal stages, based on our previous de novo-assembled genome of S. peregrina. Totals of 2457, 3656, 3764, and 2554 differentially expressed genes were identified. The specific genes encoding the structural constituent of cuticle were significantly differentially expressed, suggesting that degradation and synthesis of cuticle-related proteins might actively occur during metamorphosis. Molting (20-hydroxyecdysone, 20E) and juvenile (JH) hormone pathways were significantly enriched, and gene expression levels changed in a dynamic pattern during the developmental stages. In addition, the genes in the oxidative phosphorylation pathway were significantly expressed at a high level during the larval stage, and down-regulated from the wandering to pupal stages. Weighted gene co-expression correlation network analysis (WGCNA) further demonstrated the potential regulation mechanism of tyrosine metabolism in the process of puparium tanning. Moreover, 10 consistently up-regulated genes were further validated by qRT-PCR. The utility of the models was then examined in a blind study, indicating the ability to predict larval development. The developmental, stage-specific gene profiles suggest novel molecular markers for age prediction of forensically important flies. [ABSTRACT FROM AUTHOR]

Summary: We collected and identified beetles belonging to different species of Coleoptera that are of forensic importance in India by employing cytochrome oxidase subunit II. An attempt was made to create a molecular database of the reference sequences, as most of the studied species were not available in online databases. Sequences obtained from the present work will contribute to the creation of a national database, allowing for an increase in the knowledge of Coleoptera species of forensic interest. The foremost concern in forensic entomology is the explicit identification of the species recovered from the crime scene. From the different orders of insects, Diptera is the prime focus in this field, followed by Coleoptera, whose identification can be extremely helpful for corpses in later decomposition stages. In this study, cytochrome oxidase subunit II (COII) was used to check its adequacy as a genetic marker and to create a reference database for eleven species belonging to five families of Coleoptera, namely, Silphidae, Staphylinidae, Histeridae, Dermestidae and Scarabaeidae, from two different states in India to assist in the accurate identification of imperative beetle species in medico-legal entomology. To achieve this, standard protocols of DNA isolation, amplification and sequencing were followed. We concluded that the COII gene can be used as a molecular marker for the identification of forensically relevant species, as observed from the similarities between the phylogenetic relationship constructed by COII and morphological data. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Applied Entomology is the property of Chinese Journal of Applied Entomology, Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Environmental pressures challenge the survivorship of wildlife, and adaptive ability is key to their success and establishment. However, it remains unclear how changes in environmental conditions affect the phenotypic plasticity of some species. Studies that consider the effect of the environment (e.g., climate changes) on species morphology are essential to better understand his potential resilience. Thus, Diptera species appear as an excellent model. Here we investigated the effect of environmental conditions on wing size and shape within and between one native and one invasive blowfly species populations, along an ecoregional gradient in Brazil. For native species, wing size was affected by almost all environmental variables, while wing shape was unaffected. For invasive species, humidity best explained both wing size and wing shape variation. Our results show a differentiation between populations for both species, but a transversal comparison reveals that invasive species do not seem to be affected in the same way as the native species by climatic variations in the field. Temperature and humidity are the most significant environmental variables associated with differences in wing size and shape for invasive species, indicating an elevated climate tolerance of this species, which may be associated with its invasive characteristics and adaptive potential. Thus, our findings shed light on the importance of assessing the individual and compound effect of environmental variation and biological invasion on the adaptation of native species, since it is expected to increase 3 °C with a 20% overall reduction in the amount of rainfall in the tropical region of Brazil by 2100. [ABSTRACT FROM AUTHOR]

Calliphora vicina Robineau-Desvoidy is a primary colonizing species of decomposing bodies, being predominant in the cold seasons in Buenos Aires province, Argentina. The minimum post-mortem interval (PMImin) was estimated using growth and larval length parameters as the accurate entomological evidence of the elapsed time since death. Specimens were reared at 12, 14, 16, and 18 °C, and the larval and pupal growth rates were determined. Differences in the developmental times were registered (43.2 days at 12 °C, 34.3 days at 14 °C, 29.1 days at 16 °C, and 24.3 days at 18 °C), with the lifespan shortened at higher temperatures, without changing the larval length range (-1.5--18.0 mm). Temperature also influenced the survival and size of emerged adults; the highest percentage of survival registered at 14 (29.73%) and 16 °C (38.65%), and more mortality in extreme temperatures (19.00% at 12 °C and 12.61% at 18 °C). Adults reared at 12 °C were the largest of all, females were longer than males, and body size showed an inverse relationship with survival. These results allowed the PMI estimation based on data from local populations instead of consulting papers based on other regions. [ABSTRACT FROM AUTHOR]

Background: Mortality among children under 5 years is an important health indicator. Therefore, determining the most common causes and manners of death according to the postmortem data is necessary for designing intervention programs to reduce mortality. Objectives: This study aimed to evaluate the causes and manners of death in children aged under 5 years old in Tehran, Iran using autopsy findings. Methods: This descriptive cross-sectional study was conducted on the data of all deaths among children aged under 5 years who were referred to the Legal Medicine Organization of Tehran, Iran, during January 2009-December 2019. The data were collected using the checklists of demographic characteristics, autopsy, toxicological findings, pathological findings, hospital records, and judicial documents, which were then analyzed. Results: Among 1750 children aged under 5 years old included in this study, 898 (51.3%) cases were male, and 997 (56.9%) were hospitalized. Most of the mortality cases occurred about two months after birth. The most common causes of death were found as congenital cardiovascular anomalies (14.7%), pneumonia (11.7%), and preterm labor (11%). Moreover, natural death (77.7%), accidental death (17.7%), homicide (2.7%), and unknown death (2%) were the major manners of death in these children. Conclusions: Postmortem examination to determine the causes of unnatural death could help clinicians and policymakers to propose a suitable intervention for reducing the mortality rate in children under 5 years. [ABSTRACT FROM AUTHOR]

Background: The contributions of forensic entomology and arthropod in legal investigations have been known for centuries. There are many ways that insects can be used to help in solving a crime, but the primary purpose of forensic entomology is the determination of postmortem interval. The aim of this work is to evaluate the effect of scorpion venom on the decomposition of rabbits’ carcasses and on arthropods’ colonization of rabbits’ carcasses in indoor and outdoor environments. Methods: Twenty male rabbits were included in the study, divided into four groups, with 5 rabbits in each group. two groups were killed by spinal cord separation; group (1) kept in an indoor environment and group (3) in an outdoor environment. The other two groups were killed by scorpion stings (groups 2 and 4), kept indoors, and outdoor respectively. The carcasses were allowed to decompose, and arthropods were collected daily and examined morphologically. Results: The decomposition findings were less prominent in groups 2 and 4 with less maggot mass compared to the control groups. Furthermore, arthropod species were different and showed morphological changes in the form of dryness with the formation of ulcers in the larvae, appearance of irregular groves and tunnels in insects and dryness of the outer layer with damage to the ends of beetles. Conclusion: Scorpion venom affected the decomposition rate of rabbits’ carcasses and arthropods’ colonization with the difference in morphological appearance of arthropods between the control groups and scorpion stinging groups, which may be of forensic importance. [ABSTRACT FROM AUTHOR]

Simple Summary: Sarcosaprophagous beetles are significant pests, but they are also key indicator insects for postmortem interval estimation. In this paper, the development of Necrobia ruficollis (Fabricius, 1775) at five constant temperatures between 22 °C and 34 °C was investigated. The developmental threshold temperature and thermal summation constant were estimated, the relationship between the larval body length with development time, and the widths of the head capsules and the distance between the urogomphi at different instars were studied. The results provide important basic developmental data for using N. ruficollis to estimate the minimum postmortem interval. Necrobia ruficollis (Fabricius, 1775) (Coleoptera: Cleridae) is an important cosmopolitan storage pest, and also frequently appears on highly decomposed and skeletonized corpses. It is a forensically important species expected to indicate a longer postmortem interval (PMI). Therefore, we investigated the development of N. ruficollis at five constant temperatures between 22 °C and 34 °C. Under temperatures of 22, 25, 28, 31, and 34 °C, the mean (±SD) developmental durations from eggs to adults were 93.00 ± 1.63, 70.67 ± 0.94, 65.33 ± 3.40, 47.33 ± 0.94, and 56.66 ± 8.73 days, respectively. According to the developmental time and accumulated degree hours results, an isomorphen diagram and thermal summation model were generated. The calculated values of developmental threshold temperature and accumulated temperature constant were estimated by a linear model to be 14.51 ± 0.52 °C and 684.12 ± 33.85 degree days, respectively. Lower developmental thresholds, intrinsic optimum temperature, and upper lethal developmental threshold temperature were estimated by a nonlinear model to be 14.61, 25.90, and 34.94 °C. Morphological indexes of larvae were obtained by in vivo measurements. A growth curve and an equation of the relationship between development time and body length were simulated. In addition, the widths of the head capsules and the distance between the urogomphi of larvae at different instars were determined by cluster analysis. Classifiers were created and validated by linear discriminant analysis. These results provide important basic developmental data for using N. ruficollis to estimate the minimum postmortem interval (minimum PMI). However, this study was only conducted under constant temperature, and the applicability of these data to variable temperature conditions needs to be further confirmed. [ABSTRACT FROM AUTHOR]

Accurate age estimates of immature necrophagous insects associated with a human or animal body can provide evidence of how long the body has been dead. These estimates are based on species-specific details of the insects’ aging processes, and therefore require accurate species identification and developmental stage estimation. Many professionals who produce or use identified organisms as forensic evidence have little training in taxonomy or metrology, and appreciate the availability of formalized principles and standards for biological identification. Taxonomic identifications are usually most readily and economically made using categorical and qualitative morphological characters, but it may be necessary to use less convenient and potentially more ambiguous characters that are continuous and quantitative if two candidate species are closely related, or if identifying developmental stages within a species. Characters should be selected by criteria such as taxonomic specificity and metrological repeatability and relative error. We propose such a hierarchical framework, critique various measurements of immature insects, and suggest some standard approaches to determine the reliability of organismal identifications and measurements in estimating postmortem intervals. Relevant criteria for good characters include high repeatability (including low scope for ambiguity or parallax effects), pronounced discreteness, and small relative error in measurements. These same principles apply to individuation of unique objects in general.Key points Metrological rigour can increase in forensic entomology by selecting measurements based on their metrological qualities. Selection of high-quality features for morphological identification of organisms should consider these criteria: (1) pronounced discreteness of features (minimising group overlap or maximizing interval); (2) high repeatability of assessment (such as symmetrical width rather than asymmetrical length); (3) small relative error in measurement (selecting the physically largest continuous rigid feature for measurement). These metrological principles also apply to individuation of unique objects in general.

In forensic entomology the estimation of the minimum postmortem interval and any other evaluation depend on the accuracy of the species identification. Despite the most common species received a lot of attention in the last years, other species belonging to less common families still remain poorly studied and sometimes their immature stages are not described. The finding of Physyphora alceae (Preyssler, 1791) (Diptera, Ulidiidae) in a decapitated cadaver in Northern Italy allowed the description of the III instar larvae and of the puparium of this species. Further molecular analysis allowed the confirmation of the correct species identification and the monophyly of the genus. In general, phylogenetic analysis confirmed the monophyly of the family, however some issues arised on the real monophyly of the genus Homalocephala. This paper highlights the need of a combination of morphological and molecular identification of the species especially when immatures are collected.