Estimating the preappearance interval from temperature in Creophilus maxillosus L. (Coleoptera: Staphylinidae)

Abstract: Some carrion insects appear on carcasses late in decomposition. While using these late‐arriving taxa to estimate postmortem interval, it is vital to accurately estimate an interval preceding their appearance on a cadaver called the preappearance interval (PAI). This paper tests the possibilities of a PAI estimation from temperature in case of late‐arriving, predatory carrion beetle Creophilus maxillosus. The relationship between temperature and PAI of adult and larval C. maxillosus was modeled using the results of a large‐scale experiment on succession in forests of Central Europe. In both stages, PAI was found to be strongly, inversely related to temperature averaged for the duration of PAI. The PAI estimation from mean degree‐day accumulations (mDDA), estimated DDA, and “PAI*T” functions were tested using external successional data. The greatest accuracy of PAI estimates was achieved using an exponential “PAI*T” function.     

Electrophysiological and Behavioral Responses of Thanatophilus sinuatus Fabricius (Coleoptera: Silphidae) to Selected Cadaveric Volatile Organic Compounds

Soon after death, carcasses release volatile chemicals that attract carrion insects including Silphidae. Nevertheless, it is not known which chemical cues are involved in the attractiveness of the carcass. So far, little information is available on the chemical ecology of carrion beetles, particularly concerning the subfamily of Silphinae. The biological role of selected cadaveric volatile organic compounds including dimethyldisulfide (DMDS), butan‐1‐ol, n‐butanoic acid, indole, phenol, p‐cresol, putrescine, and cadaverine on the silphine species, Thanatophilus sinuatus Fabricius, was investigated using both electrophysiological and behavioral techniques. Among the tested cadaveric compounds, butan‐1‐ol and DMDS elicited the strongest electroantennography (EAG) from both T. sinuatus male and female antennae. In a two‐arm olfactometer, males and females were significantly attracted to DMDS for both tested doses, whereas only males were attracted to p‐cresol at 100 ng. Putrescine was repellent to males at the dose of 1 μg.     

Diel Activity and Effect of Carcass Decomposition on the Attractiveness to the Forensically Important Species Oxelytrum Discicolle (Coleoptera: Silphidae)†

Oxelytrum discicolle is a carrion beetle commonly found in the Neotropical region, mainly associated with cadavers and carcasses. Information on behavior related to the likelihood of cadaver colonization is scarce. We performed two field experiments in peri‐urban forest in Brazil in order to strengthen our knowledge on O. discicolle behavior. In the first experiment, we assessed the preference for piglet carcasses at different stages of decomposition offered simultaneously in two seasons, dry and rainy, while in the second experiment we investigated the diel activity of adults. A total of 493 adults were collected in the experiments. A significantly higher abundance was detected in the rainy season. There was a marked preference for carcasses at advanced stages. Adults exhibited a typical nocturnal behavior, the number trapped at night was 17 times greater than those collected during the day. Oxelytrum discicolle is nocturnal species that prefers carcasses in advanced stage of decomposition.     

Dermestes maculatus (Coleoptera: Dermestidae) development under fluoxetine effect using two drug administration models

Fluoxetine is a selective serotonin reuptake inhibitor, commonly used for the treatment of a variety of psychopathological conditions. As such, fluoxetine may be expected to appear in clinical and forensic cases. Dermestes maculatus De Geer (Coleoptera: Dermestidae) has been recognized as a relevant component of the insect fauna associated with decomposing human and animal remains. Experiments were conducted to study the effect of fluoxetine on developing D. maculatus using two‐drug administration models: a non‐living animal model (pork muscle) and a living one (Sus scrofa L. pigs). We assessed the duration of immature stages and total life cycle, as well as morphological parameters (body length, cephalic width, and weight). The effect of fluoxetine was studied at an overdose concentration: In the non‐living animal model the drug was mixed with macerated pork muscle (2000 mg/kg) and in the living animal model, pigs were given the drug orally (833 mg/kg). A control was used for each model. Daily observations were performed from the beginning to the end of the experiments. GC‐MS was used for drug detection and quantification. There were no statistically significant differences in the duration of immature stages, life cycle, larval mortality, morphological parameters, or sex ratio, between treatment and control, regardless of the drug administration model. Given that fluoxetine had no detectable effect on the development of D. maculatus, detection of this drug in forensic situations would not compromise the accuracy of PMI estimations.     

Decomposition and Arthropod Succession in Whitehorse,Yukon Territory,Canada

Forensic arthropod succession patterns are known to vary between regions. However, the northern habitats of the globe have been largely left unstudied. Three pig carcasses were studied outdoors in Whitehorse, Yukon Territory. Adult and immature insects were collected for identification and comparison. The dominant Diptera and Coleoptera species at all carcasses were Protophormia terraneovae (R‐D) (Fam: Calliphoridae) and Thanatophilus lapponicus (Herbst) (Fam: Silphidae), respectively. Rate of decomposition, patterns of Diptera and Coleoptera succession, and species dominance were shown to differ from previous studies in temperate regions, particularly as P. terraenovae showed complete dominance among blowfly species. Rate of decomposition through the first four stages was generally slow, and the last stage of decomposition was not observed at any carcass due to time constraints. It is concluded that biogeoclimatic range has a significant effect on insect presence and rate of decomposition, making it an important factor to consider when calculating a postmortem interval.     

Analysis of the Effect of Cyclophosphamide and Methotrexate on Chrysomya megacephala (Diptera: Calliphoridae),

Forensic entomotoxicology investigates the effects of chemical substances in the development of scavenger insects and the reflection on estimating the minimum postmortem interval (PMI). To evaluate the impact of the chemotherapeutic drugs cyclophosphamide (CF) and methotrexate (MTX) on the postembryonic development of Chrysomya megacephala, controlled experiments were performed by rearing the larvae on spiked minced beef with different concentrations of these drugs. The results indicated that CF significantly decreased their developmental rate up to 28 h but had no significant effect on larval and adult sizes, survival rate, and sex ratio, whereas MTX decreased larval and adult sizes, survival rate, and there was a deviation in the expected sex ratio toward females in MTX‐exposed larvae but had no significant impact on developmental rate. These negative interference factors should be considered in cases of suspected death of people that have undergone chemotherapy.     

Host‐habitat Location by the Parasitoid,Nasonia vitripennis Walker (Hymenoptera: Pteromalidae)

This study investigated the role of odorant cues used during host‐habitat location by the generalist parasitoid, Nasonia vitripennis Walker. Nasonia vitripennis is a common parasitoid of Dipteran pupae found in association with decaying carrion. Behavioral assays were used to investigate the host‐habitat searching behavior under different scenarios. First, we demonstrated N. vitripennis to be significantly attracted toward odorant cues associated with decaying meat. The biological activity of nine of the volatile molecules constituting the odor of decaying meat was tested on the searching behavior of parasitoid females through two complementary chemoecological approaches: electroantennography (EAG) and olfactometry bioassays. Butanoic acid and butan‐1‐ol elicited high olfactory responses, but no attraction was induced by these two chemicals. Behavioral assays showed that, among the VOCs tested, methyldisulfanylmethane (DMDS) was the only volatile chemical to induce attraction in N. vitripennis.     

The Influence of Preburial Insect Access on the Decomposition Rate

Abstract: This study compared total body score (TBS) in buried remains (35 cm depth) with and without insect access prior to burial. Sixty rabbit carcasses were exhumed at 50 accumulated degree day (ADD) intervals. Weight loss, TBS, intra‐abdominal decomposition, carcass/soil interface temperature, and below‐carcass soil pH were recorded and analyzed. Results showed significant differences (p << 0.001) in decomposition rates between carcasses with and without insect access prior to burial. An approximately 30% enhanced decomposition rate with insects was observed. TBS was the most valid tool in postmortem interval (PMI) estimation. All other variables showed only weak relationships to decomposition stages, adding little value to PMI estimation. Although progress in estimating the PMI for surface remains has been made, no previous studies have accomplished this for buried remains. This study builds a framework to which further comparable studies can contribute, to produce predictive models for PMI estimation in buried human remains.     

Estimating the pre-appearance interval from temperature in Necrodes littoralis L. (Coleoptera: Silphidae)

While estimating post-mortem interval from entomological evidence it is useful to divide it into two separate intervals: the pre-appearance interval (PAI) and the development interval in the case of immature insects and PAI and the presence interval in the case of adult insects. An accurate estimate of PAI is critically important particularly in the case of those insects which appear on remains late in decomposition. In this paper the possibilities of PAI estimation from temperature are evaluated in late-arriving, forensically important carrion beetle Necrodes littoralis. The relationship between PAI of adult and larval N. littoralis and temperature was modelled by using results of succession experiments in forests of Central Europe. In both stages PAI was found to be strongly, inversely dependent on mean daily ground level temperatures averaged for the duration of PAI. Several methods of PAI estimation were proposed and their accuracy was initially evaluated with external successional data.     

Molecular Identification of Necrophagous Dermestes Species in South Korea Using Cytochrome c Oxidase Subunit I Nucleotide Sequences (Genus Dermestes)

Species identification of necrophagous insects found on a dead body is an essential key in applying medicolegal entomology to the estimation of postmortem interval (PMI). Due to limited morphological identification of insect evidence, several studies have identified species using molecular information such as DNA markers. While considerable cytochrome c oxidase subunit I (COI) gene sequence data of necrophagous fly species have been collected and annotated, those of necrophagous beetle species have not. Since necrophagous beetles such as Dermestes species have a larval period longer than that of flies, beetles are useful in even the late decomposition phase in estimating minimum PMI. To obtain the full-length COI gene sequences of six Dermestes species collected from South Korea, we designed primers for polymerase chain reaction amplification and sequencing. The obtained full COI nucleotide sequences were used for performing phylogenic analysis and comparison with previously reported sequences. The results demonstrated that the COI gene sequences could be used to identify forensically important Dermestes species in South Korea.     

A Forensic Entomological Analysis Can Yield an Estimate of Postmortem Interval,and Not Just a Minimum Postmortem Interval: An Explanation and Illustration Using a Case

Several authors claimed that a forensic entomological analysis can never be interpreted as the postmortem interval (PMI), but that it can be interpreted as a minimum postmortem interval (PMI_min), or dead for not less than X amount of time. Because in most instances, a PMI estimate should be a range, that is, the set of values from maximum postmortem interval (PMI_max) to PMI_min, objections to PMI estimation are objections to PMI_max estimation. The arguments for this position did not address the substantial body of literature describing estimation of both PMI_max and PMI_min using insect succession analysis. This report reviews the scientific issues and presents a recent homicide investigation in which several forensic entomology experts used the absence of carrion insects on the corpse, a kind of succession analysis, to estimate PMI or PMI_max.     

Growth of Chrysomya megacephala (Fabricius) Maggots in a Morgue Cooler*

Abstract: In estimating the postmortem interval (PMI) using maggots obtained during autopsy, the forensic entomologist makes decisions regarding the effects of low‐temperature storage of the body on the insects. In this case report, a corpse was found in an abandoned house in the residential area of Bukit Mertajam, Penang, Malaysia. The maggots were found to be alive inside the mouth of the deceased although the corpse had been in the morgue cooler for 12 days. The maggots were reared and identified as Chrysomya megacephala (Fabricius). The emerged adult flies were kept as a stock colony, and the duration of development under the indoor fluctuating temperature regime was studied. The total duration of developmental process of this species was 9.5 ± 0.5 days, and the PMI estimated was 3.2 ± 0.6 days. This case report demonstrates the survival of Ch. megacephala maggots for 12 days and their growth inside the morgue cooler.     

Using Frons Width to Differentiate Blow Fly Species (Diptera: Calliphoridae) Phormia regina (Meigen) and Protophormia terraenovae (Robineau‐Desvoidy)

Protophormia terraenovae (Robineau‐Desvoidy) (Diptera: Calliphoridae) and Phormia regina (Meigen) (Diptera: Calliphoridae) are morphologically similar blow fly species commonly used for estimating postmortem intervals. Field collection and storage of adults can result in color changes, in particular on calypters and palps; often collected specimens show damage such as wing fray or fungal growth. We measured the frons width: total head width ratio using photographs (ImageJ version 1.49) to differentiate these two species. Both sexes were distinguishable to species, with the greatest difference between males: 12.34% P. terraenovae versus 1.62% P. regina, less so for females: 40.25% P. terraenovae, versus 33.65% P. regina. Incorporating this feature into future blow fly keys would help with distinguishing field‐caught specimens when other features are obstructed.     

Diversity of Forensic Rove Beetles (Coleoptera,Staphylinidae) Associated with Decaying Pig Carcass in a Forest Biotope

Most forensic studies are focused on Diptera pattern colonization while neglecting Coleoptera succession. So far, little information is available on the postmortem colonization by beetles and the decomposition process they initiate under temperate biogeoclimatic countries. These beetles have, however, been referred to as being part of the entomofaunal colonization of a dead body. Forensic entomologists need increased databases detailing the distribution, ecology, and phenology of necrophagous insects, including staphylinids (Coleoptera, Staphylinidae). While pig carcasses are commonly used in forensic entomology studies to surrogate human decomposition and to investigate the entomofaunal succession, very few works have been conducted in Europe on large carcasses. Our work reports the monitoring of the presence of adult rove beetles (Coleoptera, Staphylinidae) on decaying pig carcasses in a forest biotope during four seasons (spring, summer, fall, and winter). A total of 23 genera comprising 60 species of rove beetles were collected from pig carcasses.     

Influence of Paraquat on Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) Infesting Minced‐beef Substrates in Kelantan,Malaysia

This study investigated the influence of paraquat, a prevalent poison used by suicides, on initial oviposition and development of Chrysomya megacephala (Fabricius) using minced‐beef substrates. Paraquat in lethal dose for human (40 mg/kg), two times the lethal dose (80 mg/kg) and five times the lethal dose (200 mg/kg) were mixed thoroughly with respective minced‐beef substrates (1 kg each) that were decomposed in a shaded habitat fully protected from rain. Results of four replications of the above experiment revealed that the presence of paraquat neither delayed initial oviposition nor prolonged the developmental stages of C. megacephala. Therefore, estimation of postmortem interval (PMI) based on empirical baseline data obtained using animal models devoid of any poisons would still be appropriate for estimating PMI in paraquat‐related deaths.     

DEET (N,N‐Diethyl‐meta‐toluamide) Induced Delay of Blowfly Landing and Oviposition Rates on Treated Pig Carrion (Sus scrofa L.)

Abstract: The question of whether the insect repellent N,N‐Diethyl‐meta‐toluamide (DEET) affected fly attraction, oviposition, and larval development was investigated; in part, to determine whether the common habit of wearing DEET as a repellent could affect the rate of human decomposition. Experiments using pig surrogates of human decedents were carried out in a rural environment. Dead piglets were sprayed with DEET, and fly behavior, colonization levels, and maggot development were compared with those in nonsprayed controls. Piglets treated with DEET experienced significant delays in fly visitation and oviposition and delayed appearance of each larval instar, as well as reduced total larval numbers (p < 0.01 for all variables), with subsequently reduced decomposition (p < 0.05). Such changes in fly behavior and larval population development would significantly impact the estimation of the period following the death from entomological evidence in decedents wearing DEET at the time of their death.     

A Preliminary Forensic Entomological Study of Beetles (Coleoptera) in Interior Alaska,USA*

Forensic entomology uses knowledge of arthropod ecology to help solve crimes. There has been no published forensic entomological research in Alaska. We used one piglet carcass split in half to create two carcass plots in Fairbanks (~64.8°N, subarctic) that were sampled over a period of 59 days in 2019. Four pitfall traps were placed around each carcass, and four similarly arranged pitfall traps were placed 40 m distant as controls. Traps were emptied approximately weekly covering the first four stages of decomposition. We focused on adults of the larger-bodied (>9 mm) families and subfamilies of Coleoptera: Staphylinidae (subfamily Staphylininae), Carabidae, and Silphidae. A total of 621 specimens were collected and processed: 29 staphylinines, 210 carabids, and 382 silphids. A one-way ANOVA showed no significant difference between the mean numbers of staphylinines or carabids caught in carcass versus control traps. Silphids showed significantly greater mean number of beetles caught in carcass traps relative to control traps. Four species of Silphidae were documented, but contrary to similar studies, the vast majority of specimens belonged to two species of Nicrophorus (N. vespilloides Herbst and N. investigator Zetterstedt). Each of the three target taxa showed a peak in the number of specimens collected during the bloat stage of decomposition despite the carabid peak being driven by a phytophagous species.     

Sex Determination from the Frontal Bone: A Geometric Morphometric Study

Sex estimation in human skeletal remains when using the cranium through traditional methods is a fundamental pillar in human identification; however, it may be possible to incur in a margin of error due because of the state of preservation in incomplete or fragmented remains. The aim of this investigation was sex estimation through the geometric morphometric analysis of the frontal bone. The sample employed 60 lateral radiographs of adult subjects of both sexes (30 males and 30 females), aged between 18 and 40 years, with mean age for males of 28 ± 4 and 30 ± 6 years for females. Thin‐plate splines evidenced strong expansion of the glabellar region in males and contraction in females. No significant differences were found between sexes with respect to size. The findings suggest differences in shape and size in the glabellar region, besides reaffirming the use of geometric morphometrics as a quantitative method in sex estimation.     

深圳地区夏季室内外猪尸体上嗜尸性昆虫的群落演替

目的：根据深圳地区夏季室内外猪尸体上主要嗜尸性昆虫的生长发育和群落演替规律来推断死亡时间（PMI）。方法从2013年5月初到8月初,在深圳市公安局法医检验中心模拟的室内、外场地分别放置成年猪尸体两头,观察尸体腐败的过程和猪尸体上出现主要昆虫的种类,记录猪尸体腐败各阶段的历期,以及主要昆虫在尸体上的群落演替。结果室内、外的猪尸体分别经历了412.5 h和325 h进入白骨化阶段。主要的嗜尸性昆虫,蝇类以大头金蝇、绯颜裸金蝇、星岛金蝇,甲虫类以大隐翅甲、赤颈郭公甲、丽腐阎甲、白腹皮蠹为优势代表性种类。室外猪尸体在较多雨水的影响下,优势蝇群明显产生二代,室内的猪尸体上未见较大规模二代产生。结论深圳地区夏季室内外猪尸体上主要昆虫的群落演替有规律性,7种代表性昆虫及其幼虫整体的活动规律对推断PMI有较大的意义。