Development of Phormia regina at seven constant temperatures for minimum postmortem interval estimation.

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.

Characterization of insect stains produced by Dermestes maculatus De Greer (Coleoptera: Dermestidae) resulting from interactions with human blood.

Insect stains produced by adult Dermestes maculatus were characterized during interactions with human blood. Beetles were offered wet or dried blood positioned on ceramic tiles under laboratory conditions. Despite a life history strategy geared toward consumption of dried food stuffs, adult beetles interacted with wet blood more frequently than dried and produced more insect stains after ingesting wet blood. Most (> 95%) of the insect stains produced were the result of fecal elimination. These stains varied in morphologies but were consistently tan/light, black/grey, or red in color; were round to amorphous in shape; and frequently possessed tails. Tailed stains typically were tadpole-shaped or long and tapering from the stain body, yielding Ltl/Lb ratios greater than 1. Tails were the result of beetle locomotion while defecating. Human blood was detected in defecatory stains when using ABA Hematrace® lateral flow assays. When beetles interacted with dried blood, the bloodstains were most often modified due to physical disruption rather than feeding activity. This yielded flaking or dislodgement of the original stains. Within a forensic context, it is unknown whether D. maculatus interacts with any type of bloodstains at a crime scene.

First record of Diamesus osculans (Vigors, 1825) (Coleoptera: Silphidae) colonization on a human corpse.

A partially skeletonized human corpse was found in bushes in Selangor, Malaysia in June 2020. Entomological evidence was collected during the autopsy and sent to the Department of Medical Microbiology and Parasitology, Faculty of Medicine, Universiti Teknologi MARA (UiTM) for minimum postmortem interval (PMImin) analysis. Standard protocols were applied when processing preserved and live insect specimens of both larval and pupal stages. Entomological evidence revealed that the corpse was colonized by Chrysomya nigripes Aubertin, 1932 (Diptera: Calliphoridae) and Diamesus osculans (Vigors, 1825) (Coleoptera: Silphidae). Chrysomya nigripes was chosen as the PMImin indicator as this fly species is an earlier colonizer compared to D. osculans beetle larvae which their presence is the indicative of late stage of decomposition. For the present case, the pupae of C. nigripes were the oldest insect evidence collected and based on the available developmental data, the estimated minimum PMI was established between 9 and 12 days. It is noteworthy to highlight that this is the first record of D. osculans colonization on a human corpse.

Effects of population variations and temperature on Chrysomya megacephala (Diptera: Calliphoridae) development: implications for estimating the postmortem interval.

Forensic entomology requires knowledge of the developmental rates of the species that colonize a body after death to estimate the postmortem interval (PMI). These developmental rates may vary depending not only on the species but also on the geographic location due to population differences. Therefore, the objectives of this work were to determine the developmental duration of the forensically important fly Chrysomya megacephala under constant controlled and field condition temperatures and to compare these results, through a meta-analysis, with data reported by other authors on populations from different localities. For this, C. megacephala colonies were established in the laboratory, and the duration of the life cycle was studied at two controlled temperatures (25 °C and 27 °C) and field conditions (27.5 ± 3.2 °C). Analysis of variance was performed to determine differences in developmental time and larval length between constant laboratory temperatures and field conditions. A generalized linear model was performed with predictor variables extracted from the literature (diet, relative humidity, latitude, longitude) to evaluate the effect of population variation on developmental times. The results showed significant differences in developmental times between 25 and 27 °C. As expected, the complete life cycle of C. megacephala was shorter at 27 °C. Finally, the meta-analysis suggested differences between the developmental times of different populations, based on temperature and geographic location. The results of this study provide fundamental developmental data to use C. megacephala in PMI estimations. Finally, we suggest that, when making expert reports, information from local populations should be used to determine a more accurate and reliable PMI.

Development of Megaselia scalaris at constant temperatures and its significance in estimating the time of death.

Megaselia scalaris (Schmitz, 1938) (Diptera: Phoridae) is a common insect in forensic science that is frequently found in indoor cases, particularly on corpses in closed environments. Although this species is useful for estimating the minimum postmortem interval (PMImin) in the absence of Calliphoridae, there is a lack of data on its development in China. Herein, we studied the development of M. scalaris exposed to seven constant temperatures ranging from 16 to 34 °C. The mean (± SD) developmental durations of M. scalaris from egg to adult stage at 16, 19, 22, 25, 28, 31, and 34 °C were 1486.9 ± 75.3, 823.7 ± 42.8, 448.2 ± 59.8, 417.7 ± 19.7, 297.2 ± 27.3, 272.9 ± 10.4, and 253.0 ± 5.0 h, respectively. The mean (± SE) lower developmental threshold temperature (TL) and the thermal summation constant (K) were determined by a linear model as 12.69 ± 0.3 °C and 4965.8 ± 227.9-degree hours, respectively. A nonlinear model estimated the lower developmental threshold temperature, intrinsic optimum temperature, and upper lethal developmental threshold temperature as 14.58, 21.00, and 34.15 °C, respectively. We established three development models to estimate the age of the immature insect, namely the isomegalen diagram, isomorphen diagram, and thermal summation model. In addition, a regression analysis of the relationship between body length and total development time from hatching to pupariation was performed. Our findings provide a basis for applications of M. scalaris in PMImin estimations.

Bionomics, reproductive traits and assessment of forensic relevance of Peckia (Peckia) chrysostoma (Wiedemann, 1830) (Diptera: Sarcophagidae).

Peckia (Peckia) chrysostoma (Wiedemann, 1830) (Diptera: Sarcophagidae) is a colonizer of cadavers in the Neotropical Region. Nevertheless, data on development for the P. (P.) chrysostoma (e.g., instar duration) and behavioral strategies used by the species for locating and colonizing a corpse are scant. We aimed to explore bionomic and reproductive aspects of the flesh fly P. (P.) chrysostoma, and in this article we: (a) provide quantitative data on the life cycle of P. (P.) chrysostoma; (b) present bionomic measurements (length and weight) of larvae and pupae; (c) describe intrauterine egg and larvae development; and (d) analyze the ovo/larviposition behavior by gravid females. Females showed ovaries with discernible eggs and larvae between 8 and 10 days (x̅ = 23.3 eggs/female). This study reports the first observation of egg deposition, an atypical behavior for the species. The average development time for immature stages was 22.24 h and 21.36 h for 1st and 2nd respectively, and 3rd showed an average development time of 80.47 h. Pupa had the longest duration (x̅ = 295.69 h). A direct increase was observed in weight (P < 0.05) and length (P < 0.05) throughout time. The average survival time of males and females is approximately 30 days. This study expands the knowledge on P. (P.) chrysostoma, such as facultative ovoviviparity under laboratory conditions and the life cycle, which may benefit future studies for accuracy in entomology-based estimation of minimum post-mortem interval (min PMI).

Variations in cuticular hydrocarbons of Calliphora vicina (Diptera: Calliphoridae) empty puparia: Insights for estimating late postmortem intervals.

Necrophagous flies, particularly blowflies, serve as vital indicators in forensic entomology and ecological studies, contributing to minimum postmortem interval estimations and environmental monitoring. The study investigates variations in the predominant cuticular hydrocarbons (CHCs) viz. n-C25, n-C27, n-C28, and n-C29 of empty puparia of Calliphora vicina Robineau-Desvoidy, 1830, (Diptera: Calliphoridae) across diverse environmental conditions, including burial, above-ground and indoor settings, over 90 days. Notable trends include a significant decrease in n-C25 concentrations in buried and above-ground conditions over time, while n-C27 concentrations decline in buried and above-ground conditions but remain stable indoors. Burial conditions show significant declines in n-C27 and n-C29 concentrations over time, indicating environmental influences. Conversely, above-ground conditions exhibit uniform declines in all hydrocarbons. Indoor conditions remain relatively stable, with weak correlations between weathering time and CHC concentrations. Additionally, machine learning techniques, specifically Extreme Gradient Boosting (XGBoost), are employed for age estimation of empty puparia, yielding accurate predictions across different outdoor and indoor conditions. These findings highlight the subtle responses of CHC profiles to environmental stimuli, underscoring the importance of considering environmental factors in forensic entomology and ecological research. The study advances the understanding of insect remnant degradation processes and their forensic implications. Furthermore, integrating machine learning with entomological expertise offers standardized methodologies for age determination, enhancing the reliability of entomological evidence in legal contexts and paving the way for future research and development.

Methods for the optimal preservation of blow fly intra-puparial forms for morphological analysis in forensic casework.

Accurate minimum post-mortem interval (minPMI) estimations often rely on a precise age determination of insect developmental stages, which is significantly influenced by environmental temperature. An optimal preservation of the entomological samples collected at crime scenes is pivotal for a reliable aging of immature insect samples. For blow flies (Diptera: Calliphoridae), the most widely used insect indicators in forensic investigations, an appropriate preservation of tissues is particularly important in the case of puparial samples because aging methods for intra-puparial forms usually depend on morphological analyses; however, although informative soft tissues and structures could be discoloured and/or distorted if they are not properly fixed, there is a lack of studies to assess different methods for the optimal preservation of intra-puparial forms collected in forensic investigations. The present study compares three preservation methods for intra-puparial forms of the blow fly Calliphora vicina Robineau-Desvoidy, 1830: (i) direct immersion into 80% ethanol, (ii) puncturing of the puparium and hot water killing (HWK) prior to preservation in 80% ethanol, and (iii) HWK without puncturing before preservation in 80% ethanol. External and internal morphological analyses of intra-puparial forms of different ages were conducted to assess the quality of preservation. The results indicate that direct immersion in ethanol led to poor preservation, affecting both external and internal tissues. Both methods with HWK resulted in a better preservation, but puncturing resulted, in some cases, in physical damage of the specimens. HWK without puncturing emerged as the optimal preservation method, consistently yielding high preservation scores for both external and internal morphological analyses. These findings have practical implications for forensic practitioners and emphasise the need for updating some published guidelines and protocols in forensic entomology.

Establishment of a multiplex PCR-CE assay for the simultaneous and rapid analysis of age markers for Calliphora vicina pupae.

Insects, especially blow flies, are forensically relevant to determine the minimal postmortem interval (PMImin), based on the fact that they are usually the first colonisers of a body. By estimating the age of immature blow flies, interferences can be made about the time since death. Whilst morphological parameters are valuable for age estimation of blow fly larvae, gene expression profiling is more applicable for blow fly pupae. Here, the age-dependent changes in the gene expression levels during the development are analysed. 28 temperature-independent markers have already been described for the age estimation of pupae of the forensically important blow fly Calliphora vicina and are analysed by RT-qPCR. To allow simultaneous analysis of these age markers, a multiplex assay was developed in the present study. After reverse transcription, the markers are analysed simultaneously in an endpoint PCR and subsequently separated by capillary electrophoresis (CE). This method is highly attractive due to its quick and easy procedure and interpretation. The present age prediction tool was adapted and validated. The multiplex PCR assay reproduced the same expression profiles as the RT-qPCR assay based on the same markers. The statistical evaluation shows that the new assay has a lower precision but a better trueness for age determination compared to the RT-qPCR assay. Since the new assay is also qualified to estimate the age of C. vicina pupae and is practical, cost-effective and, even more importantly, time-saving, it is attractive for use in forensic casework.

Parasitic and predatory behavior of Alysia manducator (Hymenoptera: Braconidae) on blow fly larvae feeding on an adult pig carcass in the Western Cape Province of South Africa: preliminary observations and forensic implications.

Wasps are part of the entomofauna associated with vertebrate carrion. They are known to parasitize and prey on specific life stages of insect hosts such as eggs, larvae, pupae, and/or adults associated with vertebrate carrion. However, reports of parasitic behavior of wasps on carrion-associated insect life stages and their possible forensic implications are non-existent in the Western Cape Province of South Africa. This study is part of ongoing research investigating the entomofauna and their pattern of succession on an adult pig carcass in Cape Town, South Africa. During this study, the parasitic wasp Alysia manducator was noted parasitizing and preying on blow fly larvae associated with the decomposing carcass. The arrival of A. manducator coincides with the occurrence of blow fly eggs and/or larvae on the carcass. These wasps were seen in close association with the eggs and larvae of blow flies on various parts of the carcass and some wasps were seen dragging fly larvae attached to their ovipositors away from one part of the carcass to another. Some A. manducator were also observed walking over several larvae on the carcass while exhibiting a stabbing behavior presumably in search of a host for oviposition. We suggest that the observations recorded in this study are of considerable forensic importance as the dragging effect and predatory and stabbing behavior exhibited by A. manducator could potentially disrupt the feeding and development of the fly larvae on the carcass. This could subsequently alter the process of carcass decomposition and/or affect minimum post-mortem interval estimations.

An effective method for preparing histological sections of blow fly intra-puparial stages for minimum PMI estimations.

Blow flies (Diptera: Calliphoridae) are generally early colonisers of fresh cadavers, enabling the estimation of a minimum post-mortem interval (minPMI) based on an accurate aging of the oldest immature stages associated with a cadaver. In blow flies, the pupal stage and the subsequent development of the adult take place inside a protective case, the puparium, formed from the hardened and darkened cuticle of the third instar larva. Because the puparium is an opaque structure that shows virtually no external changes, qualitative analyses of the internal tissues can be very informative for determining reliable age-specific morphological markers. Those analyses can be performed using either non-invasive but expensive and not widely accessible techniques, or traditional histological methods, which are invasive as they require the serial sectioning of the sample. Histological methods are often readily available for forensic researchers and practitioners; however, the histological study of blow fly intra-puparial stages has traditionally been hampered by the poor paraffin infiltration of tissues due to the abundance of fat bodies, resulting in usually fragmented sections and the subsequent loss of relevant information. We present here an effective method for the preparation of histological sections of blow fly intra-puparial stages, maximising the paraffin infiltration while enabling the production of clean and entire sections that allow for the use of reliable age-specific morphological markers, thus improving the accuracy of minPMI estimations when access to more costly techniques is not feasible.

Survival of Calliphora vicina (Diptera: Calliphoridae) embryos under cold temperature conditions: forensic implications.

Most blow flies (Diptera: Calliphoridae) species are sarcosaprophagous during the larval stage, primarily feeding on the soft tissues of carcasses during the early stages of decomposition, making them valuable forensic indicators for minimum post-mortem interval (minPMI) estimations. Like other insects, their developmental rates are strongly influenced by the environmental temperature. Although several studies have examined the influence of temperature on the development of different blow fly species, the impact of cold temperatures remains largely unstudied, despite its potential forensic implications. The present study investigates the effect of three cold temperatures (0, -2.5 and -5°C) on the survival of Calliphora vicina embryos of five different ages (0%, 20%, 40%, 60% and 80% of the total embryonic development) and two exposure times (6 and 24 h). Our results revealed significant differences in egg survival at the earliest embryonic stages (0% and 20% of the total embryonic development), resulting in high mortality rates. While at 20% of the total embryonic development high mortality was only observed under -5°C, at 0% of the total embryonic development high mortality rates were observed at all the temperatures tested. Although C. vicina embryos demonstrate tolerance to cold temperatures once they have completed the first 20% of the total embryonic development, potentially mitigating the impact of cold weather events, the possibility of minPMI underestimations due to the death of the first egg batches should not be disregarded. Additionally, considering that the embryonic development stages may last for several days under low temperatures, caution should be taken in the analysis of entomological evidence if a cadaver is discovered following cold weather episodes.

A methodological approach to age estimation of the intra-puparial period of the forensically relevant blow fly Calliphora vicina via Fourier transform infrared spectroscopy.

Estimating the age of immature blow flies is of great importance for forensic entomology. However, no gold-standard technique for an accurate determination of the intra-puparial age has yet been established. Fourier transform infrared (FTIR) spectroscopy is a method to (bio-)chemically characterise material based on the absorbance of electromagnetic energy by functional groups of molecules. In recent years, it also has become a powerful tool in forensic and life sciences, as it is a fast and cost-effective way to characterise all kinds of material and biological traces. This study is the first to collect developmental reference data on the changes in absorption spectra during the intra-puparial period of the forensically important blow fly Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae). Calliphora vicina was reared at constant 20°C and 25°C and specimens were killed every other day throughout their intra-puparial development. In order to investigate which part yields the highest detectable differences in absorption spectra throughout the intra-puparial development, each specimen was divided into two different subsamples: the pupal body and the former cuticle of the third instar, that is, the puparium. Absorption spectra were collected with a FTIR spectrometer coupled to an attenuated total reflection (ATR) unit. Classification accuracies of different wavenumber regions with two machine learning models, i.e., random forests (RF) and support vector machines (SVMs), were tested. The best age predictions for both temperature settings and machine learning models were obtained by using the full spectral range from 3700 to 600 cm-1. While SVMs resulted in better accuracies for C. vicina reared at 20°C, RFs performed almost as good as SVMs for data obtained from 25°C. In terms of sample type, the pupal body gave smoother spectra and usually better classification accuracies than the puparia. This study shows that FTIR spectroscopy is a promising technique in forensic entomology to support the estimation of the minimum post-mortem interval (PMImin), by estimating the age of a given insect specimen.

Novel use of a servosphere to study apodous insects: Investigation of blow fly post-feeding larval dispersal.

Blow flies (Diptera: Calliphoridae) are arguably the most important providers of an estimate of minimum post-mortem interval in forensic investigations. They usually undergo a post-feeding dispersal from the body. While previous studies have looked at dispersal of groups of larvae, recording the dispersal activity of individual larvae has not previously been demonstrated. A servosphere was used here to record the speed, directionality and phototaxis of individual post-feeding larvae of two species of blow fly on a smooth plastic surface over time. The servosphere rotates to compensate for the movement of an insect placed at its apex, thereby enabling its unimpeded locomotion in any direction to be studied and behavioural changes to external stimuli recorded. To our knowledge, the servosphere has not previously been used to study apodous insects. The objective of our study was to compare dispersal behaviour of Calliphora vicina Robineau-Desvoidy and Protophormia terraenovae (Robineau-Desvoidy), both common primary colonisers of human and animal cadavers, but showing different post-feeding dispersal strategies. Larvae of C. vicina generally disperse from the body while those of P. terraenovae remain on or close to the body. Our aims were to study (1) changes in dispersal speed over a 1-h period; (2) changes in dispersal speed once a day for 4 days, between the end of feeding and onset of pupariation; and (3) response of dispersing larvae to light. We demonstrated that (1) the movement of three C. vicina larvae tracked for 1 continuous hour on 1 day slowed from an average of 3 to <1.7 mms-1; (2) the average speed of 20 larvae of C. vicina (4.08 mms-1) recorded for 5 min once per day over a 4-day period between onset of dispersal and pupariation was significantly greater than that of P. terraenovae (2.36 mms-1; p < 0.0001), but that speed of both species increased slightly over the 4 days; (3) the responses of larvae of C. vicina to changes in light direction from the four cardinal directions of the compass, showed that they exhibited a strong negative phototactic response within 5 s, turning to move at approximately 180° away from the new light position. While conducted to observe larval calliphorid post-feeding behaviour, the results of this proof of concept study show that apodous insects can be studied on a servosphere to produce both qualitative and quantitative data.

Post-mortem interval determinations using insects collected from illegally hunted and dehorned rhinoceros in the Republic of South Africa from 2014 to 2021.

Wildlife forensic science is a growing research field globally with application in criminal cases of illegal hunting requiring an estimate of time of death based on insect fauna. The techniques and procedures of forensic entomology acquired over the last 40 years, used in legal cases relating to human remains, can be adapted to decomposing wildlife. Research on carrion utilising the rate of development of insect immatures provides a biological clock from which a minimum post-mortem interval (minPMI) can be derived. The following study concerns 19 rhinoceros that were illegally killed and dehorned in the Republic of South Africa between 2014 and 2021. The paper details 74 samples of insect evidence collected from these rhinoceros remains from which an accurate estimate of their PMI was calculated. The specimens comprised 18 species from 12 families belonging to three insect orders. Many Dipteran and Coleopteran species were found on and around each carcass. The species of fly larvae (family Calliphoridae) used in each case to estimate the PMI are as follows: Chrysomya marginalis (Wiedemann) (13 cases), Chrysomya chloropyga (Wiedemann) (2 cases), Chrysomya albiceps (Wiedemann) (1 case) and Chrysomya megacephala (Fabricius) (1 case). Two species of Coleoptera from the family Dermestidae and Silphidae involved Dermestes maculatus (DeGeer) and Thanatophilus micans (Fabricius), respectively, also were involved in one PMI estimation each. The paper highlights opportunities for improving our global understanding of gaps in procedures and training related to wildlife criminal cases.

First description of immature stages and notes on the biology of the blow fly, Calliphora lopesi Mello, 1962.

Several species of the worldwide distributed genus Calliphora Robineau-Desvoidy (Insecta, Diptera, Calliphoridae) are medically important vectors and agents of myiasis. Furthermore, these flies are relevant in forensics because they are found in corpses. Information regarding the taxonomy, bionomics and distribution of Calliphora species endemic to South America, including Calliphora lopesi Mello, is scarce. To expand knowledge on C. lopesi, this study presents descriptions of eggs, larvae, puparia and developmental data at 14, 17, 20, 23 and 26 ± 1°C for the first time. Adult flies were collected from the field and kept in the laboratory to obtain samples for morphological and biological studies. Immatures were examined using light and scanning electron microscopy. To assess the growth rate, 10 specimens from each temperature group were randomly removed from the diet and weighed every 24 h from larval hatching until pupation. The minimum developmental threshold, thermal constant and linear development-rate equations were calculated for each stage. Considering weight gain records and survival rates, the optimum temperature for the development of C. lopesi ranges from 23 to 26°C. A key to third-instar larvae of known Neotropical species of Calliphora was also provided to assist in identification. The information provided in this study should be useful in expanding knowledge about Neotropical Calliphoridae species of forensic importance.

Várias espécies do gênero Calliphora Robineau­Desvoidy (Insecta, Diptera, Calliphoridae), distribuídas mundialmente, são vetores e agentes causadoras de miíases clinicamente importantes. Além disso, tais moscas são relevantes no âmbito forense porque são encontradas em cadáveres. Informações sobre a taxonomia, bionomia e distribuição de espécies de Calliphora endêmicas da América do Sul, incluindo Calliphora lopesi Mello, ainda são escassas. Para ampliar o conhecimento sobre a espécie C. lopesi, este estudo apresenta pela primeira vez as descrições de ovos, larvas e pupários, assim como dados sobre o desenvolvimento de imaturos a 14, 17, 20, 23 e 26 ± 1°C. Moscas adultas foram coletadas no campo e mantidas em laboratório para obtenção de amostras para estudos morfológicos e biológicos. Os imaturos foram examinados utilizando microscopia óptica e eletrônica de varredura. Para avaliar a taxa de crescimento, 10 espécimes de cada grupo de temperatura foram retirados aleatoriamente do substrato em que se alimentavam e pesados a cada 24 horas, desde a eclosão das larvas até a pupariação. O limiar mínimo de desenvolvimento, a constante térmica e as equações lineares da taxa de desenvolvimento foram calculados para cada estágio. Levando em consideração os registros de ganho de peso e taxas de sobrevivência, a temperatura ótima para o desenvolvimento de C. lopesi varia de 23 a 26°C. Uma chave para larvas de terceiro estádio de espécies neotropicais conhecidas de Calliphora também está sendo disponibilizada para auxiliar na identificação. Espera­se que as informações fornecidas neste estudo possam ser úteis para ampliar o conhecimento sobre espécies Neotropicais de Calliphoridae de importância forense.

A preliminary study of carrion insects in Greece and their attraction to three animal baits: a forensic entomology perspective.

The current study provides data on the composition of the dipteran and coleopteran fauna attracted to three different animal baits (chicken stomach, beef liver, fish) in the urban ecosystem of Athens, Greece. An additional objective was to examine the effect of bait choice by the collected taxa. Three trap locations were established within the Agricultural University of Athens. At each location two different types of baited traps were used to capture and record the local dipteran and coleopteran biodiversity. During the sampling period, a total of 2416 dipteran and coleopteran specimens representing 43 species were collected and identified. The three most abundant dipteran taxa were Sarcophaga sp. (Diptera: Sarcophagidae), Calliphora vicina Robineau-Desvoidy, Lucilia sericata (Meigen) (Diptera: Calliphoridae). The results showed that the structure of the local insect community associated with the decomposition of animal baits depends on the bait used for sampling. This study reinforced the feasibility of using animal tissues as attractants to a wide diversity of medical, parasitological and forensically important taxa. Overall, beef liver attracted the most insects with Diptera preferring beef liver and Coleoptera preferring chicken stomach. Furthermore, this study revealed that baits of one animal tissue did not attract all species but in combination a much better understanding of this biodiversity was achieved. Finally, from a forensic entomology perspective this study highlights the lack of understanding of the value of entomological data related to case work in Greece.

New geographic location data on the occurrence and abundance of carrion insects of forensic interest.

The world's forensic entomologists have much in common. They face similar research challenges, apply the same scientific methodology, study the same kinds of evidence, and access global research databases. Nevertheless, some regional heterogeneity inevitably exists. For most countries, and particularly those that have complex and diverse ecosystems, the current priority is to stimulate use of forensic entomology by establishing open access databases with time series data using standardised protocols for occurrence, abundance, distribution, niche preferences, life cycle, and identification characteristics for the key regional species that may be encountered in forensic entomology cases. Even in countries where forensic entomology is routinely used as a tool in reconstructing the history of corpses found on crime scenes in accordance with principles found in the rapidly developing body of literature, there is still much to learn about forensically useful insects. Examples of regional gaps include the taxonomy of lesser-known carrion insect species, seasonal occurrence of carrion species in the country's various geographical regions, and rates of development of the local species modeled in the many site situations of forensic interest. The first published study of carrion insects found in Athens, Greece published in this journal is an example of entomologists in a region taking the necessary first step towards establishing baseline data about native and introduced species and hence, physiological and behavioural responses to local environmental conditions, including life cycles and likelihood of occurrence or absence in the region.

Decomposition and insect succession on human cadavers in a humid, continental (Dfb) climate (Quebec, Canada).

Forensic entomology has been developing globally for decades. Despite this discipline being used in criminal investigations around the world, only a few controlled studies have been performed on human cadavers in human decomposition facilities, with the majority of these being conducted in warm and often dry climates. Therefore, the purpose of our research was to catalogue the first published data on insects associated with decomposed human bodies in a humid, continental (Dfb) climate. Specifically, our objective was to document the diversity and succession of the entomofauna associated with human cadavers throughout the decomposition process, in the Quebec province of Canada, during the summer season. Two human cadavers were studied in 2020 at the site for Research in Experimental and Social Thanatology, REST, located in Bécancour (Quebec, Canada). Insects (and other arthropods) were regularly sampled by visual observations, collection from the cadavers, and by using an entomological net and pitfall traps. Our results highlight that the decomposition process is a heterogeneous and complex process in Quebec, with cadavers showing signs of precocious desiccation/mummification. In addition, our observations confirm that the presence of superficial skin lesions accelerates the colonization of blow flies (Diptera: Calliphoridae) and, consequently, the process of decomposition. Finally, we were able to discriminate between "early colonizers" (e.g., Calliphoridae Lucilia sp. or Calliphora livida), "late colonizers" (e.g., larvae of Piophilidae or Heleomyzidae), and "non-specific colonizers." We also officially report the first observation of Cochliomyia macellaria (Diptera: Calliphoridae) in Quebec. These findings will provide new information to help medico-legal death investigations by determining the minimum time elapsed since death and the circumstances surrounding death.

Initial laboratory validation of temperature development models for Necrodes littoralis L. (Staphylinidae: Silphinae).

Development models of necrophagous insects are applied in forensic entomology for post-mortem interval estimation. Such estimates may be used as scientific evidence in legal investigations. For this reason, it is important that the models are valid and that the expert witness is aware of their limitations. Necrodes littoralis L. (Staphylinidae: Silphinae) is a necrophagous beetle species that frequently colonizes human cadavers. Temperature models of development for the Central European population of these beetles were recently published. In this article, we present results of the laboratory validation study for these models. Errors of beetle age estimation differed significantly between the models. Thermal summation models yielded the most accurate estimates, and the isomegalen diagram least accurate estimates. Errors of the beetle age estimation varied across beetle developmental stages and rearing temperatures. In general, most development models of N. littoralis were satisfactorily accurate in estimating beetle age under laboratory conditions; therefore, the study provides initial evidence to support their validity in forensic cases.