Applying Knowledge of Species‐Typical Scavenging Behavior to the Search and Recovery of Mammalian Skeletal Remains

Forensic investigations involving animal scavenging of human remains require a physical search of the scene and surrounding areas. However, there is currently no standard procedure in the U.K. for physical searches of scavenged human remains. The Winthrop and grid search methods used by police specialist searchers for scavenged remains were examined through the use of mock red fox (Vulpes vulpes) scatter scenes. Forty‐two police specialist searchers from two different regions within the U.K. were divided between those briefed and not briefed with fox‐typical scavenging information. Briefing searchers with scavenging information significantly affected the recovery of scattered bones (χ² = 11.45, df = 1, p = 0.001). Searchers briefed with scavenging information were 2.05 times more likely to recover bones. Adaptions to search methods used by searchers were evident on a regional level, such that searchers more accustom to a peri‐urban to rural region recovered a higher percentage of scattered bones (58.33%, n = 84).     

Forensic Imaging‐Guided Recovery of Nuclear DNA from the Spinal Cord*,†

Abstract: Our objective is to document the recovery of DNA from the spinal cord or surrounding dura mater in 11 cases of severely burned human remains. Radiographs established that portions of charred tissue contained spine segments. Multidetector computed tomography (MDCT) revealed that each spine specimen contained an intact spinal cord remnant. A full DNA profile was obtained from seven specimens using spinal cord dura mater in six specimens and spinal cord medulla in one specimen. A partial profile was obtained from four specimens (spinal cord dura mater, 2; spinal cord medulla, 2). Bone and muscle surrounding the spinal cord appear to insulate nucleic acid containing tissue from critical thermal degradation. The spinal cord, which is easily identified by MDCT examination of remains and easily recovered at the postmortem examination, can be a source of DNA with extraction yields comparable with other tissue sources. Specimens of dura mater are preferable as processing time is faster than bone.     

Scanning Electron Microscopy–Energy‐Dispersive X‐Ray (SEM/EDX): A Rapid Diagnostic Tool to Aid the Identification of Burnt Bone and Contested Cremains

This study investigates the use of Scanning electron microscopy–energy‐dispersive X‐ray (SEM‐EDX) as a diagnostic tool for the determination of the osseous origin of samples subjected to different temperatures. Sheep (Ovis aries) ribs of two experimental groups (fleshed and defleshed) were burned at temperatures of between 100°C and 1100°C in 100°C increments and subsequently analyzed with the SEM‐EDX to determine the atomic percentage of present elements. Three‐factor ANOVA analysis showed that neither the exposure temperature, nor whether the burning occurred with or without soft tissue present had any significant influence on the bone's overall elemental makeup (p > 0.05). The Ca/P ratio remained in the osseous typical range of between 1.6 and 2.58 in all analyzed samples. This demonstrates that even faced with high temperatures, the overall gross elemental content and atomic percentage of elements in bone remain stable, creating a unique “fingerprint” for osseous material, even after exposure to extreme conditions.     

Determining Postmortem Interval Using Glycoproteinous Adhesion Deposits by Balanus improvisus on Human Skeletal and Dental Remains

An anthropological analysis was conducted on skeletal and dental remains brought to the Galveston County Medical Examiner's office. The skeletal remains were dry, fragmented, and absent of typical fluvial characteristics. During microscopic examination, semitransparent, circular objects were discovered on the dentition, the mandible, tibial plateau, and distal femur. The objects were glycoproteinous adhesions deposited by the acorn barnacle, Balanus improvisus. B. improvisus is an intertidal barnacle found in estuaries in Galveston Bay. Basal diameter of the adhesions on the dentition were significantly smaller than those found on the postcranial bones (p = 0.010), indicating two consecutive cohorts adhered to the bone and dentition. As settlement typically occurs once a year, this would indicate that the remains were in the fluvial environment for at least 375–410 days. It is important in geographic areas that have prevalent fluvial environments that human remains, particularly dentition, are microscopically examined for marine life evidence.     

Elemental Analysis of Variably Contaminated Cremains Using X‐ray Fluorescence Spectrometry

Analyzing and identifying skeletal remains becomes increasingly difficult when remains have been cremated, especially in cases where the cremated material may have been intentionally contaminated with nonskeletal material. This study examined the potential of X‐ray fluorescence spectrometry (XRF) to detect the presence of nonskeletal contaminants in samples of cremains. Eleven samples of cremains were variably combined with concrete mix and analyzed using XRF. Photon counts of elements in each sample were analyzed, and the coefficient of determination (R²) using unweighted linear regression as a function of percent cremains was calculated. Results showed that with changes in the proportion of skeletal material and contaminant, there were significant (R² > 0.90) changes in detected levels of phosphorus, potassium, zinc, aluminum, and sulfur. The use of XRF is concluded to be a valid approach in the identification of the presence of nonskeletal material in potentially contaminated cremains.     

Gender Determination of Adult Individuals by Three‐Dimensional Modeling of Canines

Gender determination is a fundamental issue in forensic anthropology. Many techniques based on bone and dental remains have been proposed. It is not always possible to implement the techniques using bones, but teeth are often perfectly preserved. It has been demonstrated that the canine has the greatest sexual dimorphism, and the aim of this work was to provide an easy and accurate dental technique for determining the gender in the absence of other skeletal elements. The sample was composed of 210 CT scans with four healthy canines. The 840 canines were modeled using MIMICS^® 10.01 software. The total volume of each tooth was determined. Seven mathematical models were determined by binary logistic regressions and ranked in order of relative performance. The seven proposed predictive models thus performed (0.910 ≤ AUC ≤ 0.938), with overall rates of correct predictions between 82.38 and 85.24%. The 4‐canine model is the most powerful for predicting the gender.     

X‐ray Scattering Evaluation of Ultrastructural Changes in Human Dental Tissues with Thermal Treatment

Micro‐ and ultrastructural analysis of burned skeletal remains is crucial for obtaining a reliable estimation of cremation temperature. Earlier studies mainly focused on heat‐induced changes in bone tissue, while this study extends this research to human dental tissues using a novel quantitative analytical approach. Twelve tooth sections were burned at 400–900°C (30‐min exposure, increments of 100°C). Subsequent combined small‐ and wide‐angle X‐ray scattering (SAXS/WAXS) experiments were performed at the Diamond Light Source synchrotron facility, where 28 scattering patterns were collected within each tooth section. In comparison with the control sample, an increase in mean crystal thickness was found in burned dentine (2.8‐fold) and enamel (1.4‐fold), however at a smaller rate than reported earlier for bone tissue (5–10.7‐fold). The results provide a structural reference for traditional X‐ray scattering methods and emphasize the need to investigate bone and dental tissues separately to obtain a reliable estimation of cremation temperature.     

CT‐Guided,Minimally Invasive,Postmortem Needle Biopsy Using the B‐Rob II Needle‐Positioning Robot

CT‐guided, minimally invasive needle biopsy techniques can be used to retrieve tissue or body fluid samples for histologic tissue diagnoses in forensic investigations. The purpose of this study was an evaluation of CT‐guided needle‐positioning robot B‐Rob II. To operate under CT guidance, we adopted the B‐Rob II robotic needle placement system and its workflow. The accuracy and speed of the procedure were tested on a gelatin phantom in a series of 21 biopsies. We achieved an average needle placement accuracy of 1.8 mm (±1.1 mm) using robotic assistance. The procedure required an average of 2 min 21 s. The needle placement accuracy for minimally invasive needle biopsies using the B‐Rob II biopsy robot is sufficiently accurate and fast for forensic postmortem examinations of focal organ changes. Further tests will be performed to test the feasibility of the robot for performing biopsies of focal organ changes in human bodies.     

A Reliable Regression‐Based Approach for Reassociating Human Skeletal Elements of the Lower Limbs from Commingled Assemblages

Accurate sorting of commingled human remains comprises a fundamental requirement for all further anthropological analyses. The lower limb bones are particularly important for reconstructing biological profiles. This study introduces a metric technique for sorting these elements using eight standard anthropological measurements and 222 adult individuals from Greece. The bones utilized were the os coxae, the femora, the tibiae and the tali. Simple regression analyses were used to develop functions for reassociating articulating bones, providing strong correlations (r = 0.74–0.95, p‐value <0.05) and high coefficients of determination (r²=0.54–0.91). Blind tests demonstrated that combining metric and morphoscopic techniques provides an excellent sorting accuracy for the hip and knee joints (ten of ten individuals), allowing for a reliable reassociation between a sex and age indicator (os coxae) and a body size indicator (femur). Overall, these results indicate the high value of metric methods in sorting commingled human remains.     

Effect of Temperature on Bone Tissue: Histological Changes

The analysis of burned human remains has been of great interest among forensic anthropologists largely due to the difficulty that their recovery, classification, reconstruction, and identification present. The main purpose of this analysis is to present histological methodology for the interpretation of bones altered by thermal processes. We include analyses of the microscopic changes among bones exposed to different temperatures, with the goal of establishing categories of histological morphology in relation to fire temperature. Samples of bone (ilium) were exposed systematically to controlled temperatures. Analysis of the resulting histological changes has allowed the formation of a clear four‐stage classification of the alterations observed. This classification should prove useful in assessing bone changes in relation to temperature of exposure, particularly in cases where this temperature was previously not known.     

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.     

Dynamics of Ninhydrin‐Reactive Nitrogen and pH in Gravesoil During the Extended Postmortem Interval

Estimating the postmortem interval of skeletal remains is difficult, as few tools exist to do so. To address this problem, we conducted a field experiment to measure the chemistry of swine (Sus scrofa domesticus Linnaeus) gravesoil after 1 year and 3 years postmortem. Carcasses were placed on the soil surface of a pasture during June in a cold (Dfa) climate. Significant (p < 0.05) increases in ninhydrin‐reactive nitrogen, pH, total nitrogen, and nitrate in gravesoil were detected 1 year postmortem. Significant differences in gravesoil chemistry were not detected 3 years postmortem. These observations coincided with gaps in plant growth 1 year postmortem and the development of lush vegetation 3 years postmortem. We conclude that these phenomena can be used to assist the decision‐making process regarding the allocation of resources during the early stages of a death investigation.     

Formulae for Estimating Skeletal Height in Modern South‐East Asians

Estimating stature in human skeletal remains of Asian ancestry is problematic for forensic anthropologists due to the paucity and uncertain suitability of regression formulae. To address this issue, our study analyzed 64 individuals from a modern skeletal collection of South‐East Asian origin and developed population‐specific ordinary least squares regression formulae to estimate skeletal height from each of the long bones of the upper and lower limbs, as well as from trunk length. Results indicate that the most accurate estimates of skeletal height from a single bone (as measured by standard error of the estimate—SEE) are from tibial length in males (SEE = 2.40 cm) and from humeral length in females (SEE = 2.59 cm), followed by femoral length (SEE = 2.84 cm). When multiple elements are considered, the combination of femoral and tibial length yields the best estimates in both sexes as well as combined sex samples (male SEE = 2.40 cm; female SEE = 2.77 cm; combined sex SEE = 2.54 cm).     

A Comparative Assessment of the Chen et al. and Suchey‐Brooks Pubic Aging Methods on a North American Sample, ,

Accurately estimating the age‐at‐death of adult human skeletons is fundamental in forensic anthropology. This study evaluates the accuracy of two pubic bone age estimation methods—Chen et al. and Suchey‐Brooks. Specimens were obtained from a known collection of modern pubic bones curated at the Maricopa County Forensic Science Center in Phoenix, Arizona. A sample of 296 left male pubic bones of European ancestry was statistically evaluated via bias, absolute mean error, and intra‐ and inter‐observer error. Results indicate that the two methods are similar; the Suchey‐Brooks method is the most accurate for aging young adults (error c. 7 years), while the Revised Chen et al. method is most accurate for aging middle‐age adults (error c. 6 years). Thus, the Chen et al. method is an important contribution to forensic anthropology for aging older adult skeletal remains. There are, however, some limitations such as subjectivity and the intricate scoring system of Chen et al. method.     

Shells and Bones: A Forensic Medicine Study of the Association of Terrestrial Snail Allopeas micra with Buried Human Remains in Brazil

Little is known regarding the scavenger fauna associated with buried human corpses, particularly in clandestine burials. We report the presence of 20 shells of the terrestrial snail Allopeas micra, within hollow bones of human remains buried for 5 years, during the process of collecting DNA material. The fact that a large number of shells of A. micra had been found in the corpse and in the crime scene supports the assumption that there was no attempt to remove the corpse from the area where the crime occurred. Despite this, our observations cannot be used to estimate the postmortem interval because there is no precise knowledge about the development of this species. This is the first record of a terrestrial snail associated with a human corpse and its role in this forensic medicine case.     

Positive Dental Identification Using Tooth Anatomy and Digital Superimposition

Dental identification of unknown human remains continues to be a relevant and reliable adjunct to forensic investigations. The advent of genomic and mitochondrial DNA procedures has not displaced the practical use of dental and related osseous structures remaining after destructive incidents that can render human remains unrecognizable, severely burned, and fragmented. The ability to conclusively identify victims of accident and homicide is based on the availability of antemortem records containing substantial and unambiguous proof of dental and related osseous characteristics. This case report documents the use of digital comparative analysis of antemortem dental models and postmortem dentition, to determine a dental identification. Images of dental models were digitally analyzed using Adobe Photoshop^TM software. Individual tooth anatomy was compared between the antemortem and postmortem images. Digital superimposition techniques were also used for the comparison. With the absence of antemortem radiographs, this method proved useful to reach a positive identification in this case.     

An Investigation of Red Fox (Vulpes vulpes) and Eurasian Badger (Meles meles) Scavenging,Scattering, and Removal of Deer Remains: Forensic Implications and Applications

Within northwest Europe, especially the United Kingdom, the red fox (Vulpes vulpes) and the Eurasian Badger (Meles meles) are the largest wild scavengers capable of modifying a set of remains through scavenging. Knowledge of region-specific and species-typical scavenging behaviors of scavengers within the crime scene area and surroundings can aid in more efficient and accurate interpretations. The scavenging behaviors of captive and wild foxes and badgers were recorded and compared through actualistic methods and direct observation. The scavenging by wild foxes and badgers of surface-deposited baits and whole deer (Cervus nippon; Capreolus capreolus) in a woodland was observed and analyzed. Wild foxes were found to scavenge deer more frequently than badgers. The scavenging of deer remains by foxes was also compared with forensic cases. The scavenging pattern and recovery distances of deer and human remains scavenged by foxes were similar but were potentially affected by the condition and deposition of a body, and the presence of clothing.     

Determination of Nitrite in Whole Blood by High‐Performance Liquid Chromatography with Electrochemical Detection and a Case of Nitrite Poisoning

Although nitrite is widely used in meat processing, it is a major toxicity hazard to children and is responsible for the blue‐baby syndrome. A simple and effective method to determine nitrite in whole blood has been devised using ion chromatography with suppressed conductivity detection. The blood sample was deproteinized by adding acetonitrile and purified with mini‐cartridges to remove hydrophobic compounds, chloride ions, and metal ions. An aliquot of the filtrate was injected onto the ion chromatography. The retention time for nitrite was 13.8 min and the detection limit of nitrite in whole blood was 0.4 μmol/L. The calibration curve was linear (r² = 0.9999) over the concentration working range. The blood nitrite concentration of a victim who attempted suicide by ingesting sodium nitrite powder was determined using the present method. The basal levels for nitrite in human blood was determined with 7.1 ± 0.9 μmol/L (n = 12).     

Powdered Activated Carbon: An Alternative Approach to Genomic DNA Purification

Forensic evidence samples are routinely found as stains on various substrates, which may contain substances known to inhibit polymerase chain reaction (PCR). The goal of this study was to evaluate post‐Chelex^®100 purification using powdered activated carbon (PAC). Mock crime scene DNA extracts were analyzed using quantitative PCR and short tandem repeat (STR) profiling to test the DNA recovery and inhibitor removal using PAC with those of the Amicon^®Ultra 100K. For extracted bloodstains on soil and wood substrates, PAC and Amicon^®Ultra 100K generated similar DNA yield and quality. Moreover, the two methods significantly decreased the concentration of humic substances and tannins compared to nonpurified extracts (p < 0.001). In instances where extracts contained indigo dye (bloodstains on denim), Amicon^®Ultra 100K performed better than PAC due to improved amplifiability. Efficient adsorption of humic substances and tannins, which are common inhibitors, indicates PAC's potential application in the purification of high‐template DNA extracts.