The Red Swamp Crayfish Procambarus Clarkii (the Louisiana Crayfish) as a Particular Scavenger on a Human Corpse

An 85-year-old man was found deceased floating in an irrigation ditch 18 days after his disappearance. During crime scene investigation, specimens of the red swamp crayfish Procambarus clarkii were found in proximity of the body. The feeding activity of these nonendemic crayfishes contributed to the formation of specific injuries on the body and in the production of a large substance defects inside the corpse. The aim of this paper was to illustrate the scavenging activity of P. clarkii on a human body and highlight the potential postmortem artefacts caused by this species. This is the first report on a real case of postmortem injuries produced by P. clarkii crayfishes on a submerged human body. So far, crustaceans are not considered useful for the estimation of the minimum postmortem interval. However, the important modifications on the corpses deriving from the activity of these animals should be kept in consideration.     

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.     

Use of Cardiac Injury Markers in the Postmortem Diagnosis of Sudden Cardiac Death

In the daily practice of forensic pathology, sudden cardiac death (SCD) is a diagnostic challenge. Our aim was to determine the usefulness of blood biomarkers [creatine kinase CK‐MB, myoglobin, troponins I and T (cTn‐I and T), and lactate dehydrogenase] measured by immunoassay technique, in the postmortem diagnosis of SCD. Two groups were compared, 20 corpses with SCD and 8 controls. Statistical significance was determined by variance analysis procedures, with a post hoc Tukey multiple range test for comparison of means (p < 0.05). SCD cases showed significantly higher levels (p < 0.05) of cTn‐T and cTn‐I compared to the control group. Although only cases within the first 8 h of postmortem interval were included, and the control group consisted mainly of violent death cases, our results suggest that blood troponin levels may be useful to support a diagnosis of SCD.     

Necrophagous Caterpillars Provide Human mtDNA Evidence*

Abstract: Decomposition of large mammalian carcasses is greatly accelerated through the action of insects. Specialized feeders capable of digesting keratin and collagen found in skin, hair, and tendons and ligaments are attracted to corpses in late stages of dry decomposition and include Tinea pellionella, the casemaking clothes moth, and Tineola bisselliella, the webbing clothes moth (Lepidoptera; Tineidae). Until now, details of the caterpillar behavior as necrophagous insects were vague. Here, we detail the behavior of each species and document the incorporation of human hair into the portable larval shelters constructed by the caterpillars of T. pellionella. Hair of the decedent used as building material for caterpillar shelters provided enough starting template to amplify and sequence the HVI and HVII sections of the control region (mtDNA) of the decedent.     

Differentiation of Morphologically Similar Human Head Hairs from Two Demographically Similar Individuals Using Amino Acid Ratios,

Human hair is frequently encountered as forensic evidence and can contribute valuable information to investigators. Conventional forensic hair analyses include microscopic hair comparison (MHC) and DNA analysis. However, MHC is not supported by statistics and DNA analysis cannot always be performed. Recent studies have demonstrated that evaluation of differences in the hair proteins may offer an alternate method to these analyses. In this study, an evaluation of the amino acids present in hair was investigated as an approach to differentiate morphologically indistinguishable hair samples from two demographically similar individuals. Proteins in the hair were digested using hydrochloric acid, and the resulting amino acids were derivatized with N,O-Bis(trimethylsilyl)trifluoroacetamide (BSTFA) for analysis using gas chromatography-mass spectrometry (GC-MS). Eight derivatized amino acids were detected and quantified relative to an internal standard, L-norvaline, and used to construct twenty-eight amino acid ratios. Hair samples were collected from four areas of the head on various days over the course of one month, and no significant differences in amino acid ratios (p-value > 0.05) were observed among the areas of the head, and the ratios were consistent over the time period of this study. Additionally, fifteen of these amino acid ratios were found to be significantly different between the two individuals when compared using a two-sample t-test (p-value ≤ 0.05). These data indicate that amino acid analysis was able to differentiate two morphologically similar hair samples from different individuals and demonstrates the applicability of this method to distinguish similar hair samples when DNA analysis cannot be performed.     

Artifacts Caused by Leaf-Cutting Ants of the Genus Atta (Hymenoptera: Formicidae): Postmortem Bite Injuries and the Tearing of Clothes

Ants are one of the first insects to find an exposed cadaver and can be present during all stages of decomposition. Although these organisms are not commonly used in postmortem interval estimates, they are to be taken into account on criminal investigations involving human corpses, since they can leave bite marks that can be mistaken for antemortem or perimortem injuries, which could be misleading when ascertaining the occurrence of abuse or physical altercation during a crime. A few studies report the action of ants on human cadavers and even though leaf-cutting ants of the genus Atta are frequently encountered in succession studies that use animal carcasses, there are no records of these fungus-growing species on human corpses. Atta is a genus restricted to the New World, ranging from northern Argentina to southern United States and acts as one of the most conspicuous neotropical herbivores. In this study, we report three cases of violent death that illustrate the impact of ants, especially those of the genus Atta, in a forensic setting. We compare the patterns displayed by postmortem bite injuries caused by leaf-cutter ants and other common species with less robust mandibles. We also present the capability of Atta ants to create artifacts by cutting victim’s clothes in a crime scene, contributing to the knowledge of ant-mediated confounding factors in crime scene investigation.     

Assessment of the Role Played by N‐propanol Found in Postmortem Blood in the Discrimination Between Antemortem Consumption and Postmortem Formation of Ethanol Using Rats

This study disproves the reliability of n‐propanol as a biomarker to establish whether the ethanol found in postmortem blood is derived from antemortem ingestion or postmortem putrefactive processes. Two groups of rats were given ethanol or normal saline solution, respectively, and sacrificed 1.5 h later. After putrefaction, blood and, in a few cases, urine samples from the rats were analyzed for ethanol and n‐propanol by head‐space gas chromatography equipped with flame ionization detection. Although the concentration ratios of ethanol/n‐propanol in the postmortem blood collected from the bodies without prior alcohol consumption were expected to be <20 (as per limited case reports and previous in vitro studies), in samples from several rats that were on saline solution, this ratio was found to exceed 20. In conclusion, the concentration ratio of ethanol/n‐propanol in postmortem blood does not allow for the discernment between antemortem ingestion and the postmortem synthesis of ethanol.     

The Significance of Adrenal Hemorrhage: Undiagnosed Waterhouse‐Friderichsen Syndrome,A Case Series

A retrospective series of five cases of nontraumatic gross adrenal hemorrhage were identified in 800 consecutive forensic autopsies. All patients were males, of different ethnicities and with ages ranging from 2 to 48 years. All patients had a clinical history and autopsy findings suggestive of sepsis. Pre‐ or postmortem microbiological cultures were variably positive for Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus aureus, and methicillin‐resistant Staphylococcus aureus. The fifth case was positive for yeast and a coagulase negative staphylococcus; contamination of this culture medium cannot be excluded. No cases had a culture positive for Neisseria meningitidis. We find that the reviewed patients with grossly or microscopically identifiable adrenal hemorrhage were otherwise healthy individuals who died suddenly as a consequence of bacterial infection. In each case, signs and symptoms compatible with premortem adrenal insufficiency were reported; in no instance was the adrenal hemorrhage clinically identified.     

Forensic Features of Lethal Late‐Presenting Diaphragmatic Hernias

Diaphragmatic defects are a relatively common and benign finding in adults which may be congenital or secondarily acquired. The case files at Forensic Sciences South Australia were reviewed over a 10‐year period from July 2005 to June 2015 for all adult (>17 years) cases in which diaphragmatic hernias were identified at postmortem examination that had either caused or contributed to death. Five cases were found: age range 49–90 years (average 67.2 years); male:female ratio 2:3. Herniated organs included the stomach (N = 3), small (N = 3) and large intestines (N = 2). Mechanisms of death involved lung compression with respiratory failure and/or mediastinal shift, and vascular compromise with gastric or intestinal infarction and/or perforation. Diaphragmatic hernias may not be identified until the time of autopsy and may be quite complex entities to evaluate due to a lack of clinical history and to difficulties in determining their origin and possible contributions to mechanisms of death.     

Determination of Malathion levels and its effect on the development of Chrysomya megacephala (Fabricius) in South China

The present study determines the concentration of Malathion in rabbit tissues and Dipteral larvae feeding on those tissues. Malathion was found in all muscle and liver samples of the test rabbit, as well as larvae fed on it. Samples from the control rabbit and pupae from all colonies were negative for Malathion. Correlations were found between administered dosage, tissue concentrations and younger or prepuparial larvae. Effects of Malathion on the development rate of Chrysomya megacephala (Fabricius) were also reported. C. megacephala is the most common fly species found on corpses in South China during the early stages of decomposition. Significant differences in larval growth rate were both observed among the colonies fed on muscle and liver. The presence of Malathion in both muscle and liver appears to retard the normal growth rate of C. megacephala in larval stage. Larvae from all colonies fed on tissues from rabbits treated with Malathion were smaller and attained maximum length later than those from the control colony. Duration of the larval and pupal stages was both significantly prolonged for larvae on tissues from rabbit receiving Malathion than those from the control colony. The difference of the duration of the larval and pupal stages together from the muscle colonies would alter the postmortem interval estimation by up to 36 h. As for liver colonies, it would alter the postmortem interval estimation by up to 28 h. A significantly different duration of the larval and pupal stages from the muscle colonies would alter a postmortem estimate by up to 28 h relative to the liver colonies.     

Corneal‐Smart Phone: A novel method to intelligently estimate postmortem interval

The changes of postmortem corneal opacity are often used to roughly estimate the postmortem interval (PMI) in forensic practice. The difficulty associated with this time estimate is the lack of objective means to rapidly quantify postmortem corneal changes in crime scenes. This study constructed a data analysis model of PMI estimation and implemented an intelligent analysis system for examining the sequential changes of postmortem corneal digital images, named Corneal‐Smart Phone, which can be used to quickly estimate PMI. The smart phone was used in combination with an attachment device that provided a darkroom environment and a steady light source to capture postmortem corneal images. By segmenting the corneal pupil region images, six color features, Red (R), Green (G), Blue (B), Hue (H), Saturation (S), Brightness (V) and four texture features Contrast (CON), Correlation (COR), Angular Second Moment (ASM), and Homogeneity (HOM), were extracted and correlated with PMI model. The results indicated that CON had the highest correlation with PMI (R² = 0.983). No intra/intersubject variation in CON values were observed (p > 0.05). With the increase in ambient temperature or the decrease in humidity, the CON values were increased. PMI prediction error was <3 h within 36 h postmortem and extended to about 6–8 h after 36 h postmortem. The correct classification rate of the blind test samples was 82%. Our study provides a method that combines postmortem corneal image acquisition and digital image analysis to enable users to quickly obtain PMI 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.     

Citrate Content of Bone for Time Since Death Estimation: Results from Burials with Different Physical Characteristics and Known PMI

A recently introduced method to determine the postmortem interval (PMI) based on quantification of the citrate content in bone was applied on the temporal bones and femora of 20 individuals buried in wooden coffins (WO) and body bags (BB), respectively. Concerning known vs. calculated PMI, a significant difference between the temporal and the femur bone samples of the same individuals was observed in the BB group (p = 0.012). In contrast, differences were insignificant for the WO group (p = 0.400). Moreover, similar levels of underestimation of PMIs resulted from the analysis of the femora for both burial groups (p = 0.247). Also, there was consistently less citrate preserved in the flat temporal bones as compared to the femora, indicating that the cortical layer of the long bones should be preferentially employed for citrate‐based PMI estimations. The results call for additional research on subsurface‐buried and surface‐deposited remains to enhance the accuracy of the published PMI equation.     

Temperature‐Dependent Postmortem Changes in Human Cardiac Troponin‐T (cTnT): An Approach in Estimation of Time Since Death

Estimation of time of death is an indispensible requirement of every medico‐legal autopsy, but unfortunately, there is not a single method by which it could be determined accurately. This study focused on the temperature‐dependent postmortem degradation of cardiac troponin‐T and its association with postmortem interval (PMI) in human. The analysis involved extraction of the protein, separation by denaturing gel electrophoresis (SDS‐PAGE), and visualization by Western blot using cTnT‐specific monoclonal antibodies. The area of the bands within a lane was quantified by scanning and digitizing the image using Gel Doc (Universal Hood). The results indicate a characteristic banding pattern among human cadavers (n = 6) and a pseudo‐linear relationship between percentage of cTnT degradation and the log of the time since death (r > 0.95), which can be used to estimate the postmortem interval. The data presented demonstrate that this technique can provide an extended time range during which PMI can be more accurately estimated.     

Volatile Organic Compound Profiling from Postmortem Microbes using Gas Chromatography–Mass Spectrometry

Volatile organic compounds (VOCs) are by-products of cadaveric decomposition and are responsible for the odor associated with decomposing remains. The direct link between VOC production and individual postmortem microbes has not been well characterized experimentally. The purpose of this study was to profile VOCs released from three postmortem bacterial isolates (Bacillus subtilis, Ignatzschineria indica, I. ureiclastica) using solid-phase microextraction arrow (SPME Arrow) and gas chromatography–mass spectrometry (GC-MS). Species were inoculated in headspace vials on Standard Nutrient Agar and monitored over 5 days at 24°C. Each species exhibited a different VOC profile that included common decomposition VOCs. VOCs exhibited upward or downward temporal trends over time. Ignatzschineria indica produced a large amount of dimethyldisulfide. Other compounds of interest included alcohols, aldehydes, aromatics, and ketones. This provides foundational data to link decomposition odor with specific postmortem microbes to improve understanding of underlying mechanisms for decomposition VOC production.     

Do bone elasticity and postmortem interval affect forensic fractographic analyses?

Forensic fractographic features of bone reliably establish crack propagation in perimortem injuries. We investigated if similar fracture surface features characterize postmortem fractures. Experimentally induced peri- and postmortem fractures were used to assess if fractographic features vary as bone elasticity decreases during the postmortem interval (PMI). Thirty-seven unembalmed, defleshed human femoral shafts from males and females aged 33–81 years were fractured at varying PMIs with a drop test frame using a three-point bending setup and recorded with a high-speed camera. Vital statistics, cause of death, PMI length, temperature, humidity, collagen percentage, water loss, fracture energy, and fractography scores were recorded for each sample. Results showed that fractographic features associated with perimortem fractures were expressed in PMIs up to 40,600 accumulated degree hours (ADH), or 60 warm weather days. Hackle was the most consistently expressed feature, occurring in all fractures regardless of ADH. The most variable characteristics were wake features (78.4%) and arrest ridges (70.3%). Collagen percentage did not correlate strongly with ADH (r = −0.04, p = 0.81); however, there was a strong significant correlation between ADH and water loss (r = 0.74, p < 0.001). Multinomial logistic regression showed no association between fractographic feature expression and ADH or collagen percentage. In conclusion, forensic fractographic features reliably determine initiation and directionality of crack propagation in experimentally induced PMIs up to 40,600 ADH, demonstrating the utility of this method into the recent postmortem interval. This expression of reliable fractographic features throughout the early PMI intimates these characteristics may not be useful standalone features for discerning peri- versus postmortem fractures.     

Investigation of homicides interred in concrete--the Los Angeles experience

Decedents interred in concrete present unique problems and investigation of these deaths necessitates a team of forensic specialists. The Los Angeles County Department of Coroner has had five such deaths in the past 18 years. The buried cases needed layer-by-layer excavation to establish the time and cause of death. Metal detectors are often used in this process. X-rays of the interred remains were completed to help with locating the decedent's position in the concrete. The breaking of concrete in some of the cases required the use of a sledgehammer and later a chisel in a manner that would not damage the remains. Postmortem dismemberment was frequent in our cases. The decedents were all female or prepubescent children, and the perpetrators were closely related to the decedents. While concrete can interfere with determination of postmortem interval, it can also preserve the remains and assist with identification.     

Microscopic Pattern of Bone Fractures as an Indicator of Blast Trauma: A Pilot Study

The assessment of fractures is a key issue in forensic anthropology; however, very few studies deal with the features of fractures due to explosion in comparison with other traumatic injuries. This study focuses on fractures resulting from blast trauma and two types of blunt force trauma (manual compression and running over), applied to corpses of pigs; 163 osteons were examined within forty fractures by the transmission light microscopy. Blast lesions showed a higher percentage of fracture lines through the Haversian canal, whereas in other types of trauma, the fractures went across the inner lamellae. Significant differences between samples hit by blast energy and those runover or manually compressed were observed (p < 0.05). The frequency of pattern A is significantly higher in exploded bones than in runover and compressed. Microscopic analysis of the fracture line may provide information about the type of trauma, especially for what concerns blast trauma.     

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.     

Quantitative MRI in Isotropic Spatial Resolution for Forensic Soft Tissue Documentation. Why and How?*

Abstract: A quantification of T1, T2, and PD in high isotropic resolution was performed on corpses. Isotropic and quantified postmortem magnetic resonance (IQpmMR) enables sophisticated 3D postprocessing, such as reformatting and volume rendering. The body tissues can be characterized by the combination of these three values. The values of T1, T2, and PD were given as coordinates in a T1–T2–PD space where similar tissue voxels formed clusters. Implementing in a volume rendering software enabled color encoding of specific tissues and pathologies in 3D models of the corpse similar to computed tomography, but with distinctively more powerful soft tissue discrimination. From IQpmMR data, any image plane at any contrast weighting may be calculated or 3D color‐encoded volume rendering may be carried out. The introduced approach will enable future computer‐aided diagnosis that, e.g., checks corpses for a hemorrhage distribution based on the knowledge of its T1–T2–PD vector behavior in a high spatial resolution.