Identification of Crystals Forming on Porcine Articular Cartilage: A New Method for the Estimation of the Postmortem Interval

Articular cartilage was examined to determine its decomposition sequence and its potential for assessing the postmortem interval. Scanning electron microscopy of articular cartilage from buried porcine trotters showed the presence of microcrystals on the synovial surface. These orthorhombic pyramidal or “coffin”‐shaped crystals, appeared at 3 weeks (22 days) after interment and disappeared after 6 weeks. The disappearance of these crystals was linked to decompositional changes to the integrity of the synovial joint. The formation and disappearance of these crystals was associated with a pH change at the cartilage surface. Scanning electron microscopy–energy‐dispersive X‐ray (SEM‐EDX) analysis showed that the five main elements contained within these crystals were carbon, nitrogen, oxygen, magnesium, and phosphorous. Such elemental analysis suggested the crystals may be struvite (MgNH₄PO₄6(H₂O)). Bacteria cultured from the cartilage synovial surface produced struvite crystals when grown in suitable media and were identified by DNA analysis to be Comamonas sp.     

Potential Use of Bacterial Community Succession in Decaying Human Bone for Estimating Postmortem Interval,,

Bacteria are taphonomic agents of human decomposition, potentially useful for estimating postmortem interval (PMI) in late‐stage decomposition. Bone samples from 12 individuals and three soil samples were analyzed to assess the effects of decomposition and advancing time on bacterial communities. Results indicated that partially skeletonized remains maintained a presence of bacteria associated with the human gut, whereas bacterial composition of dry skeletal remains maintained a community profile similar to soil communities. Variation in the UniFrac distances was significantly greater between groups than within groups (p < 0.001) for the unweighted metric and not the weighted metric. The members of the bacterial communities were more similar within than between decomposition stages. The oligotrophic environment of bone relative to soft tissue and the physical protection of organic substrates may preclude bacterial blooms during the first years of skeletonization. Therefore, community membership (unweighted) may be better for estimating PMI from skeletonized remains than community structure (weighted).     

An Aquatic Decomposition Scoring Method to Potentially Predict the Postmortem Submersion Interval of Bodies Recovered from the North Sea

This study aimed to develop an aquatic decomposition scoring (ADS) method and investigated the predictive value of this method in estimating the postmortem submersion interval (PMSI) of bodies recovered from the North Sea. This method, consisting of an ADS item list and a pictorial reference atlas, showed a high interobserver agreement (Krippendorff's alpha ≥ 0.93) and hence proved to be valid. This scoring method was applied to data, collected from closed cases—cases in which the postmortal submersion interval (PMSI) was known—concerning bodies recovered from the North Sea from 1990 to 2013. Thirty‐eight cases met the inclusion criteria and were scored by quantifying the observed total aquatic decomposition score (TADS). Statistical analysis demonstrated that TADS accurately predicts the PMSI (p < 0.001), confirming that the decomposition process in the North Sea is strongly correlated to time.     

Bioelectrical Impedance as a Technique for Estimating Postmortem Interval,,

Estimation of postmortem interval (PMI) is a critical component of death investigation. A cadaver can be hypothesized to be a resistor–capacitor (RC) circuit the impedance (Z) of which changes in a quantifiable manner as the cadaver decomposes. This hypothesis was tested using bioelectrical impedance analysis (BIA) equipment to apply a current with a fixed amplitude at a single frequency to four cadavers over time and measuring two components of Z, resistance (R) and reactance (X_c). Quadratic regression analysis between Z and accumulated degree days (ADD) showed a statistically significant parabolic relationship. The parabolic relationship poses an initial challenge to the use of the method, and additional research is needed to address this issue. However, the results of the reported research support the hypothesis that Z measured using BIA has a relationship to PMI.     

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.     

Potential Forensic Application of Receptor for Advanced Glycation End Products (RAGE) as a Novel Biomarker for Estimating Postmortem Interval

Studies which estimate the postmortem interval (PMI) based on diatoms or bacterial communities have been well‐conducted. However, predicting the PMI based on microorganisms has limited accuracy. The objective of this study was to discover specific biomarkers for estimating the PMI for drowning. The in vivo experimental model consisted of rats classified into nine groups (seven drowned groups from Day 1 to Day 7 and two control groups). Their lung tissues were analyzed for the receptor for advanced glycation end products (RAGE) mRNA expression determined by qRT‐PCR. RAGE protein levels were determined by Western blotting and immunohistochemical (IHC) staining. RAGE mRNA and protein levels gradually degraded over time. Immunostaining analysis revealed that RAGE in the Day 1 sample was significantly higher than that in the Day 6 or Day 7 samples. These results indicate that RAGE in drowned lungs could be a potential biomarker for the determination of the PMI after drowning.     

Soil Fungi: Their Potential use as a Forensic Tool

As a grave is an anomalous environment and differs from its surroundings, criminal investigators employ different techniques for locating, recovering, and analyzing clandestine graves. In this study were identified the fungi found in the soil under corpses in decomposition with an aim at relating the copresence of human remains and different fungal species. Were isolated the fungi in three ways: soil washing, serial dilutions, and moist chamber growth. Dichotomomyces cejpii, Talaromyces trachyspermus, Talaromyces flavus, and Talaromyces sp. were the representative species found—with those belonging to the ammonia group, whose fungi are the first in the succession of cadaver decomposition directly in the ground. The mycobiota found at the present study area clearly differs to mycobiota identified in control sample and from previously described species for other areas of Buenos Aires Province, Argentina. Further forensic examples of this type are needed to develop fully the detailed use of mycology as a forensic tool.     

The Effects of Soil Environment on Postmortem Interval: A Macroscopic Analysis

Abstract:  Burial environment, in particular soil moisture, has a significant impact on the type, rate, and extent of bone degradation, which ultimately affects estimations of the postmortem interval (PMI). The purpose of this research is to determine the effects of soil moisture on the color, weight, condition, and texture of bone as it relates to the PMI. Bone changes occurring over two different time intervals (2 and 5 months) were examined using 120 sus scrofa leg bones. During each time interval bones were buried in two soil environments, one of which was drier than the other. The bones in both environments lost weight over time but the net weight loss was greater for bones in the higher moisture environment. There was no change in color, texture, or overall condition, indicating that 150 days is not long enough for such alterations to occur, regardless of the moisture level of the burial environment.     

Rodents as taphonomic agents: bone gnawing by brown rats and gray squirrels

Passive infrared technology was used to film diurnal and nocturnal scavenging behavior of brown rats and gray squirrels at the University of Tennessee's Anthropological Research Facility. This direct documentation demonstrated that brown rats modified fat-laden cancellous bone while gray squirrels generally gnawed the thicker bone cortices only after fats had leached away. A case study placed in a shaded portion of the Facility indicated the postmortem interval for initial gnawing by gray squirrels was slightly over 30 months. An examination of 53 human skeletons in the William M. Bass Forensic Skeletal Collection revealed that 10 cases had gnaw marks consistent with those made by gray squirrels. One of the 10 cases had been gnawed within 16 months of time-since-death, while the remaining nine had postmortem intervals >30 months. Additional observed modifications made to nonhuman bone by gray squirrels indicate that squirrel gnaw marks on bone can serve as a minimal estimate of time-since-death in a temperate environment similar to that of East Tennessee.     

The Difficult Task of Assessing Perimortem and Postmortem Fractures on the Skeleton: A Blind Text on 210 Fractures of Known Origin

The distinction between perimortem and postmortem fractures is an important challenge for forensic anthropology. Such a crucial task is presently based on macro‐morphological criteria widely accepted in the scientific community. However, several limits affect these parameters which have not yet been investigated thoroughly. This study aims at highlighting the pitfalls and errors in evaluating perimortem or postmortem fractures. Two trained forensic anthropologists were asked to classify 210 fractures of known origin in four skeletons (three victims of blunt force trauma and one natural death) as perimortem, postmortem, or dubious, twice in 6 months in order to assess intraobserver error also. Results show large errors, ranging from 14.8 to 37% for perimortem fractures and from 5.5 to 14.8% for postmortem ones; more than 80% of errors concerned trabecular bone. This supports the need for more objective and reliable criteria for a correct assessment of peri‐ and postmortem bone fractures.     

Measurement of Postmortem Pupil Size: A New Method with Excellent Reliability and Its Application to Pupil Changes in the Early Postmortem Period

Measurement of postmortem pupil width is a potential component of death time estimation. However, no standardized measurement method has been described. We analyzed a total of 71 digital images for pupil–iris ratio using the software ImageJ. Images were analyzed three times by four different examiners. In addition, serial images from 10 cases were taken between 2 and 50 h postmortem to detect spontaneous pupil changes. Intra‐ and inter‐rater reliability of the method was excellent (ICC > 0.95). The method is observer independent and yields consistent results, and images can be digitally stored and re‐evaluated. The method seems highly eligible for forensic and scientific purposes. While statistical analysis of spontaneous pupil changes revealed a significant polynomial of quartic degree for postmortem time (p = 0.001), an obvious pattern was not detected. These results do not indicate suitability of spontaneous pupil changes for forensic death time estimation, as formerly suggested.     

Specific Patterns of Canine Scavenging in Indoor Settings

Postmortem animal mutilations due to domestic dogs in isolated domestic deaths are taphonomic modifications regularly observed by forensic pathologists. They are rarely described in the literature; however, even though they present specific patterns. Through 41 cases, 10 at the forensic institute in Lille (France) and 31 at the New York City Office of Chief Medical Examiner (USA), plus 22 cases from the literature, specific locations and patterns of postmortem scavenging lesions are proposed. These lesions are mainly distributed in three locations: the face, especially the nose and the mouth (73.1%), the neck (43.1%), and the arm (shoulder/upper limb [29.2%], hand [26.8%]). We discuss the time span between death and scavenging, the consequences on identification, and comparison with outdoor settings. Outdoor scavenging lesions are mainly located on the trunk and limbs usually sparing the head, which strongly differs from indoor distribution and imply different animal motivations.     

Internal Morphological Analysis for Age Estimation of Blow Fly Pupae (Diptera: Calliphoridae) in Postmortem Interval Estimation*

Abstract: Larvae and pupae of blow fly species are frequently used in postmortem interval estimation, their age indicating minimum time since death. Most studies have considered age estimation of larvae, neglecting study of pupae. Relative development of external pupal features is useful, but there are also internal changes during metamorphosis that may be indicators of age, utilizing histological techniques. This study aimed to optimize preservation and histological analysis of blow fly pupae, specifically Calliphora vicina Robineau‐Desvoidy and Lucilia sericata (Meigen), and to examine internal features with potential for age estimation. Effect of hot‐water‐killing and different preservatives were examined. It was determined that blow fly pupae should be pierced through the three tagma, hot‐water‐killed, and preserved in 80% ethanol as the optimal preservation for subsequent analyses. Hematoxylin and eosin stained pupal sections revealed differences in brain and thoracic muscle development throughout the pupal stage with potential for age estimation.     

The Potential of High-throughput Metagenomic Sequencing of Aquatic Bacterial Communities to Estimate the Postmortem Submersion Interval

Human remains can be discovered in freshwater or marine ecosystems, circumstances where insects and other invertebrates have infrequently been used for understanding the time of postmortem submersion. In this study, the identification and succession of epinecrotic bacterial communities on vertebrate remains were described during decomposition in a temperate headwater stream during two seasons (summer and winter). Bacterial communities were characterized with 454 pyrosequencing and analyzed at phyletic and generic taxonomic resolutions. There was a significant increase in genera richness over decomposition during both seasons. Additionally, multivariate statistical modeling revealed significant differences in bacterial communities between seasons at both taxonomic resolutions and siginificant genera differences among sampling days within each season, suggesting a succession of these communities. These data are the first to describe aquatic bacterial succession using high-throughput metagenomic sequencing on vertebrate remains submerged in a freshwater habitat, and provide initial evidence for their potential use in forensic investigations.     

Predicting the Postmortem Submersion Interval for Human Remains Recovered from U.K. Waterways*

Abstract: This article aims to increase accuracy in estimating the postmortem submersion interval (PMSI) for bodies recovered from rivers in the United Kingdom. Data were collected from closed case files, crime scene reports, and autopsy files concerning bodies recovered over a 15‐year period from the River Clyde, Scotland, and the River Mersey and canals in northwest England. One hundred and eighty‐seven cases met the study criteria and were scored by quantifying the overall amount of decomposition observed in each case. Statistical analysis showed that the duration of a body’s submergence in water and the temperatures to which it was exposed, as measured in accumulated degree days (ADD), had a significant effect on the decay process. Further analysis indicated that there were no significant differences in decomposition between the waterways. By combining the data from all study samples, it was possible to produce a single linear regression model for predicting ADD from observed decomposition.     

近十年来我国现代法医昆虫学研究进展

法医昆虫学(Forensic entomology)指的是应用昆虫学知识解决法律上问题的法医学分支学科,在死后间隔时间判断,特别是腐败尸体的死后间隔时间判断等方面有著无可比拟的优越性。本文讨论了法医昆虫学的研究领域,从理论研究与实践应用两方面概述了近十年来法医昆虫学在我国的进展。     

The Effect of Body Mass on Outdoor Adult Human Decomposition

Forensic taphonomy explores factors impacting human decomposition. This study investigated the effect of body mass on the rate and pattern of adult human decomposition. Nine males and three females aged 49–95 years ranging in mass from 73 to 159 kg who were donated to the Complex for Forensic Anthropology Research between December 2012 and September 2015 were included in this study. Kelvin accumulated degree days (KADD) were used to assess the thermal energy required for subjects to reach several total body score (TBS) thresholds: early decomposition (TBS ≥6.0), TBS ≥12.5, advanced decomposition (TBS ≥19.0), TBS ≥23.0, and skeletonization (TBS ≥27.0). Results indicate no significant correlation between body mass and KADD at any TBS threshold. Body mass accounted for up to 24.0% of variation in decomposition rate depending on stage, and minor differences in decomposition pattern were observed. Body mass likely has a minimal impact on postmortem interval estimation.     

The Role of Vitreous Magnesium Quantification in Estimating the Postmortem Interval

The use of magnesium as a parameter to estimate the time of death is controversial. In 32 traumatic deaths with known postmortem intervals (PMIs), small quantities of vitreous humor (VH) were sampled repetitively every 3 h until 24 h postmortem. The bodies were kept at the constant ambient temperature of 20°C (68°F). The concentrations of magnesium were in the range of 0.47–1.15 mM. A statistically significant correlation of the concentration of magnesium with the PMI was observed (r = 0.453, p < 0.01), but with small predictive value—coefficient of variation for regression was 45.5%; the average of the paired differences between the estimated and actual PMIs was 5.24 + 3.19 h. Although useful results might be expected due to the large transmembrane gradient for magnesium, the results of this study strongly disprove the usefulness of measuring magnesium in VH to estimate the time of death.     

The formation of early stage adipocere in submerged remains: a preliminary experimental study

In some circumstances, the presence of adipocere may retard decomposition and complicate postmortem interval estimation. This article explores the correlation between Accumulated Degree Days (ADD) and early stage formation of adipocere. Sixty wild rabbit (Oryctolagus cuniculus) carcasses were used in this experiment; a control group (N = 30) deposited directly on the ground surface and an experimental group (N = 30) completely submersed in water in individual buckets. Data (water and inner body temperature, pH, and total body score) were collected every 100 ADD. Results indicated that early stage adipocere is correlated to ADD and that its formation on submersed remains is more likely to occur after 630 ADD. Skin sloughing promoted the formation of adipocere. No adipocere was formed on any of the control group rabbits. This study also highlights the fact that multiple factors influence adipocere formation and it is suggested that further research needs to be conducted into this area.