Identification of post-mortem product of zolpidem degradation by hemoglobin via the Fenton reaction.

Zolpidem, N,N-dimethyl-2-[6-methyl-2-(4-methylphenyl)imidazo[1,2-a]pyridin-3-yl]acetamide, is a hypnotic agent widely used in clinical practice but is detected in many clinical cases of fatal intoxication and suicide. In forensic toxicology, the precise determination of zolpidem concentration in blood is a must to provide concrete evidence of death by zolpidem poisoning. However, the concentrations of zolpidem in blood at autopsy often differ from those at the estimated time of death. In the present study, we found that zolpidem was degraded by hemoglobin (Hb) via the Fenton reaction at various temperatures. The mechanism underlying zolpidem degradation involved the oxidation of its linker moiety. The MS and MS/MS spectra obtained by liquid chromatography quadrupole-Orbitrap mass spectrometry (LC-Q-Orbitrap-MS) showed the formation of 2-hydroxy-N,N-dimethyl-2-(6-methyl-2-(p-tolyl)imidazo[1,2-a]pyridin-3-yl)acetamide (2-OH ZOL) in Hb/H2O2 solution incubated with zolpidem and in the blood of several individuals who died from ingestion of zolpidem. These results suggest that 2-OH ZOL is the post-mortem product of zolpidem degradation by Hb via the Fenton reaction.

Effect of humidity on postmortem changes in rats.

IMPORTANCE: In veterinary forensic science, accurately determining the postmortem interval (PMI) is crucial for identifying the causes of animal deaths. Autolysis, a significant postmortem process, influences PMI estimation, but its relationship with humidity is not well understood. OBJECTIVE: This study aimed to improve the accuracy of PMI estimates in veterinary forensic cases by looking into how different humidity levels affect autolysis in different organs of rats. METHODS: The study involved 38 male rats, examining histopathological changes in their heart, liver, and pancreas. These organs were subjected to controlled humidity levels (20%, 55%, and 80%) at a constant 22°C. Tissue samples were collected at several intervals (0 h, 12 h, 24 h, 3 days, and 8 days) for comprehensive analysis. RESULTS: Distinct autolytic characteristics in animal organs emerged under varying humidity conditions. The low-humidity environment rapidly activated autolysis more than the high-humidity environment. In addition, it was found that lower humidity caused nuclear pyknosis, cytoplasmic disintegration, and myofiber interruption. The liver, in particular, showed portal triad aggregation and hepatocyte individuation. The pancreas experienced cell fragmentation and an enlarged intracellular space. High humidity also caused the loss of striations in cardiac tissues, and the liver showed vacuolation. Under these conditions, the pancreas changed eosinophilic secretory granules. CONCLUSIONS AND RELEVANCE: The study successfully established a clear connection between the autolytic process in PMIs and relative humidity. These findings are significant for developing a more accurate and predictable method for PMI estimation in the field of veterinary forensic science.

Postmortem Diffusion of Aconitum Alkaloids and Their Metabolites in Rabbits.

OBJECTIVES: To explore the postmortem diffusion rule of Aconitum alkaloids and their metabolites in poisoned rabbits, and to provide a reference for identifying the antemortem poisoning or postmortem poisoning of Aconitum alkaloids. METHODS: Twenty-four rabbits were sacrificed by tracheal clamps. After 1 hour, the rabbits were administered with aconitine LD50 in decocting aconite root powder by intragastric administration. Then, they were placed supine and stored at 25 ℃. The biological samples from 3 randomly selected rabbits were collected including heart blood, peripheral blood, urine, heart, liver, spleen, lung and kidney tissues at 0 h, 4 h, 8 h, 12 h, 24 h, 48 h, 72 h and 96 h after intragastric administration, respectively. Aconitum alkaloids and their metabolites in the biological samples were analyzed by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). RESULTS: At 4 h after intragastric administration, Aconitum alkaloids and their metabolites could be detected in heart blood, peripheral blood and major organs, and the contents of them changed dynamically with the preservation time. The contents of Aconitum alkaloids and their metabolites were higher in the spleen, liver and lung, especially in the spleen which was closer to the stomach. The average mass fraction of benzoylmesaconine metabolized in rabbit spleen was the highest at 48 h after intragastric administration. In contrast, the contents of Aconitum alkaloids and their metabolites in kidney were all lower. Aconitum alkaloids and their metabolites were not detected in urine. CONCLUSIONS: Aconitum alkaloids and their metabolites have postmortem diffusion in poisoned rabbits, diffusing from high-content organs (stomach) to other major organs and tissues as well as the heart blood. The main mechanism is the dispersion along the concentration gradient, while urine is not affected by postmortem diffusion, which can be used as the basis for the identification of antemortem and postmortem Aconitum alkaloids poisoning.

Postmortem metabolomics: influence of time since death on the level of endogenous compounds in human femoral blood. Necessary to be considered in metabolome study planning?

INTRODUCTION: The (un)targeted analysis of endogenous compounds has gained interest in the field of forensic postmortem investigations. The blood metabolome is influenced by many factors, and postmortem specimens are considered particularly challenging due to unpredictable decomposition processes. OBJECTIVES: This study aimed to systematically investigate the influence of the time since death on endogenous compounds and its relevance in designing postmortem metabolome studies. METHODS: Femoral blood samples of 427 authentic postmortem cases, were collected at two time points after death (854 samples in total; t1: admission to the institute, 1.3-290 h; t2: autopsy, 11-478 h; median ∆t = 71 h). All samples were analyzed using an untargeted metabolome approach, and peak areas were determined for 38 compounds (acylcarnitines, amino acids, phospholipids, and others). Differences between t2 and t1 were assessed by Wilcoxon signed-ranked test (p < 0.05). Moreover, all samples (n = 854) were binned into time groups (6 h, 12 h, or 24 h intervals) and compared by Kruskal-Wallis/Dunn's multiple comparison tests (p < 0.05 each) to investigate the effect of the estimated time since death. RESULTS: Except for serine, threonine, and PC 34:1, all tested analytes revealed statistically significant changes between t1 and t2 (highest median increase 166%). Unpaired analysis of all 854 blood samples in-between groups indicated similar results. Significant differences were typically observed between blood samples collected within the first and later than 48 h after death, respectively. CONCLUSIONS: To improve the consistency of comprehensive data evaluation in postmortem metabolome studies, it seems advisable to only include specimens collected within the first 2 days after death.

LC-MS-based metabolomics reveals metabolite dynamic changes of beef after superchilling early post-mortem.

To explore the underlying mechanisms by which superchilling (SC, -3 °C within 5 h of slaughter) improves beef tenderness, an untargeted metabolomics strategy was employed. M. Longissimus lumborum (LL) muscles from twelve beef carcasses were assigned to either SC or very fast chilling (VFC, 0 °C within 5 h of slaughter) treatments, with conventional chilling (CC, 0 âˆ¼ 4 °C until 24 h post-mortem) serving as the control (6 per group). Biochemical properties and metabolites were investigated during the early post-mortem period. The results showed that the degradation of µ-calpain and caspase 3 occurred earlier in SC treated sample, which might be attributed to the accelerated accumulation of free Ca2+. The metabolomic profiles of samples from the SC and CC treatments were clearly distinguished based on partial least squares-discriminant analysis (PLS-DA) at each time point. It is noteworthy that more IMP and 4-hydroxyproline were found in the comparison between SC and CC treatments. According to the results of metabolic pathways analysis and the correlation analysis between traits related to tenderness and metabolites with significant differences (SC vs. CC), it can be suggested that the tenderization effect of the SC treatment may be related to the alteration of arginine and proline metabolism, and purine metabolism in the early post-mortem phase.

Assessing the impact of ultrasound on the rate and extent of early post-mortem glycolysis in bovine Longissimus thoracis et lumborum.

The rate of pH decline, early post-mortem, has been identified as a key factor that impacts the tenderness of meat, and manipulating this rate of pH decline is highly relevant to ensure consistent high quality meat. Ultrasound is a potential intervention in early post - mortem muscle that may have an impact on the rate of glycolysis through its ability to alter enzyme activity. Following a variety of different ultrasound treatments frequencies (25 and 45 kHz) and durations (15, 30 and 45 min), it was found, when analysed in muscle, that ultrasound treatment duration, specifically the 30 min treatment, and interaction between treatment duration and frequency, had a significant impact on the rate of pH decline, post - treatment. Frequency did not have a significant effect on the rate of pH decline, post - treatment, in muscle. Ultrasound did not have a significant permanent effect on the activity of glycolytic enzymes present in bovine Longissimus lumborum et thoracis muscle, where no significant differences were observed on the rate of pH decline and rate of change of reducing sugars, glycogen and lactic acid, when analysed in an in vitro glycolytic buffer. It seems that the impact observed in intact muscle is not as a consequence of a permanent change in enzymatic activity, instead indicating an impact on conditions in the muscle which enhanced enzyme activity.

MEDICO-LEGAL APPLICATIONS OF FRACTURE HEMATOMA: REVIEW.

This review aimed to elucidate the critical role of fracture hematoma in forensic medicine, with a specific focus on its utility in differentiating antemortem from postmortem fractures. The study seeks to provide a comprehensive synthesis of current knowledge on the subject, highlighting the biological and medico-legal implications of fracture hematoma analysis in forensic investigations. A systematic review of literature was conducted, encompassing various scientific databases including PubMed, Scopus, and Web of Science, focusing on studies published from 2000 to 2024. The search employed keywords such as "fracture hematoma," "antemortem fractures," "perimortem fractures" and "postmortem fractures," among others, to explore relevant data. Selected studies were scrutinized based on their relevance, the presence of substantial data on fracture hematoma, and their contribution to forensic analysis. The review underscores the significance of fracture hematoma as an indicator of antemortem injuries, revealing that active blood circulation at the time of injury facilitates hematoma formation. Detailed analyses within the selected studies illustrate the interplay of cellular and molecular dynamics within fracture hematomas, emphasizing the roles of cytokines, particularly IL-6, and cellular constituents in the healing process. Fracture hematoma analysis emerges as a vital forensic tool in establishing the vitality of bone fractures, enhancing the accuracy of forensic assessments. However, the review also acknowledges the challenges posed by individual healing variability and postmortem changes, suggesting a need for further research to refine the interpretative frameworks used in forensic hematoma analysis.

CT analysis of skull contents in naturally mummified human corpses, a multicentric study.

INTRODUCTION: This study aimed to assess skull contents, brain appearance, and density on postmortem computed tomography in naturally mummified corpses. MATERIAL AND METHODS: For this purpose, a retrospective multicentric study, including mummified corpses from two French centers (Brest and Nantes) and from the New Mexico Decedent Image Database (USA), was performed by analyzing postmortem computed tomography (PMCT) focused on the head and neck of partially or fully mummified corpses discovered between 2011 and 2022. The PMCT analysis provided data on the CT appearance of brains, allowing them to be classified into four different categories (desiccation, liquefaction, dura mater only (DMO), and absence), and to measure densities (HU) of the brain remains. In addition, data on postmortem intervals (PMI) from Nantes and Brest centers were collected and analyzed to test the link between brain densities and PMIs. RESULTS: 54 cases of naturally mummified corpses were included. The brains were classified as liquefied (56%), desiccated (17 %), DMO (20 %), and absent (7 %) based on their CT appearance. Dehydrated brains were significantly (p < 0.004) denser (median 102 HU, interquartile range (IQR) 41) than either liquefied brains (median 39.5 HU, IQR 9) or brains with DMO (median -25 HU, IQR 57). However, the density of brain remains was not significantly affected by where the bodies were found (p = 0,41). Analysis of PMI and brain densities was performed on 22 cases. The results showed that brain remains were significantly (p = 0.039) denser when they were found after a PMI of more than six months. CONCLUSION: Brain desiccation was the aspect with the highest densities on PMCT, and for which we were able to highlight great preservation of anatomical structures observable in living organisms.

Inhibition of pyruvate dehydrogenase accelerates anaerobic glycolysis under postmortem simulating conditions.

This research aimed to explore the potential influence of mitochondria on the rate of anaerobic glycolysis. We hypothesized that mitochondria could reduce the rate of anaerobic glycolysis and pH decline by metabolizing a portion of glycolytic pyruvate. We utilized an in vitro model and incorporated CPI-613 and Avidin to inhibit pyruvate dehydrogenase (PDH) and pyruvate carboxylase (PC), respectively. Four treatments were tested: 400 µM CPI-613, 1.5 U/ml Avidin, 400 µM CPI-613 + 1.5 U/ml Avidin, or control. Glycolytic metabolites and pH of the in vitro model were evaluated throughout a 1440-min incubation period. CPI-613-containing treatments, with or without Avidin, decreased pH levels and increased glycogen degradation and lactate accumulation compared to the control and Avidin treatments (P < 0.05), indicating increased glycolytic flux. In a different experiment, two treatments, 400 µM CPI-613 or control, were employed to track the fates of pyruvate using [13C6]glucose. CPI-613 reduced the contribution of glucose carbon to tricarboxylic acid cycle intermediates compared to control (P < 0.05). To test whether the acceleration of acidification in reactions containing CPI-613 was due to an increase in the activity of key enzymes of glycogenolysis and glycolysis, we evaluated the activities of glycogen phosphorylase, phosphofructokinase, and pyruvate kinase in the presence or absence of 400 µM CPI-613. The CPI-613 treatment did not elicit an alteration in the activity of these three enzymes. These findings indicate that inhibiting PDH increases the rate of anaerobic glycolysis and pH decline, suggesting that mitochondria are potential regulators of postmortem metabolism.

[Features of detection and interpretation of intravital and postmortem changes according to the results of traditional X-ray and X-ray computed tomography of objects from historical graves and artefacts]. / Osobennosti vyyavleniya i interpretatsii prizhiznennykh i posmertnykh izmenenii po rezul'tatam traditsionnoi rentgenografii i rentgenovskoi komp'yuternoi tomografii ob"ektov iz istoricheskikh zakhoronenii i artefaktov.

OBJECTIVE: To study emergence mechanism, physical nature, pattern of intravital and postmortem changes of biological and non-biological objects originated in the period from 1550 to 1918 yr. using traditional X-ray and X-ray computed tomography. MATERIAL AND METHODS: The relics of Saint Macarius the Roman of Novgorod, the remains of the First Reverend of the Resurrection Novodevichy Convent in Saint Petersburg Mother Superior Theophania, damages on the chair leg on which Tsesarevich Alexey sat during the shooting of Russian Emperor Nicholas II, his family and entourage in 1918 in Yekaterinburg were stidued. RESULTS AND CONCLUSION: The application of highly informative methods of traditional X-ray and X-ray computed tomography of biological and non-biological objects showed their high informativity and allowed to correctly interpret the emergence mechanism, physical nature, pattern of intravital and postmortem changes of skeleton bones and historical artefact (chair legs) originated long ago. The necessity of special professional training and advanced training of experts in forensic radiology to prevent possible diagnostic and expert errors has been substantiated.

The influence of altitude on the abundance and occurrence of species of the family Calliphoridae (Diptera) in corpses in a Mediterranean area.

A carcass is defined as a temporary resource that can support high levels of diversity compared to other resources. The level of diversity often depends on the environmental conditions in which the corpse is found. Calliphoridae (Diptera) are the most important necrophagous insects used in forensic investigations because this family is common, widespread, abundant, and usually the first to colonize a corpse. However, understanding the processes responsible for variation in calliphorid species abundance and richness along gradients in ecology remains a key challenge. In this study, we analyzed the influence of altitude on the abundance and diversity of Calliphoridae species in corpses at three different altitudes (400, 900, and 1500 m a.s.l.) in a mountainous area of southeastern Spain. The results revealed a gradient of decreasing abundance with increasing altitude. An altitudinal substitution of species was observed, with Chrysomya albiceps being found at the lowest altitude and Calliphora vicina at the highest altitude. Seasonal variation was also noted, with Ch. albiceps being the dominant species in the warmer months and C. vicina being the dominant species in winter. Our results confirm the importance of Calliphorids as seasonal, altitudinal, and environmental indicators, given the wide distribution and abundance of this family. This information is of great interest on the interpretation of data in forensic practice.

Volabolomic Fingerprinting for Post-Mortem Interval Estimation: A Novel Physiological Approach.

Death is a multifaceted process wherein each individual cell and tissue has a metabolic homeostasis and a time of functional cessation defined by the dying process as well as by intrinsic and extrinsic factors. Decomposition is physiologically associated with the release of different types of volatile organic compounds (VOCs), and these form volaboloma mortis. The main purpose of this study was to record the volabolomic fingerprint produced by volatile molecules during the physiological decomposition process of human tissue and muscle cells. The volatile chemical signature has important implications for an open issue in forensics and pathology, namely the estimation of the postmortem interval (PMI), which decreases in accuracy with the passage of time. Volatile metabolites emitted from human tissues and muscle cells at 0, 24, 48, and 72 h were recorded in real time with an electronic nose sensor device. The key findings were the continuous sampling of VOCs emitted from tissues and cells. These showed a common behavior as time progressed; particularly, after 48 h the distributions became dispersed, and after 72 h they became more variable. Volabolomic fingerprinting associated with time progression relevant to the study of PMIs was reconstructed. Additionally, there may be broader applications, such as in dog training procedures for detecting human remains, and perhaps even for studying scavenger and insect attractants.

Estimation of the post-mortem interval; added value of mechanical excitability of human skeletal muscle.

Estimation of the post-mortem interval (PMI) is mainly based on the state of body cooling, post-mortem lividity (livor mortis) and post-mortem muscle stiffness (rigor mortis). However, the time span of development of these post-mortem phenomena are influenced by a variety of factors concerning the body of the deceased and the environment in which the body is found. Subsequently, this leads to a substantial spread in upper and lower limits of PMI based on determination of the state of these phenomena. Moreover, interpretation of post-mortem phenomena like lividity, rigor and interpretation of the correction factor for Henssge's nomogram is subjective. For this reason, PMI estimations are often broad, possibly too broad to be helpful for answering questions which are relevant for the criminal investigation. Therefore, combining the outcome of different methods for estimating the PMI, the so-called compound method, is recommended. Supravital muscle reaction by mechanical stimulation of skeletal muscle is a less known aspect of the compound method. Here we present a series of cases series in which supravital muscle reaction contributed to a more precise estimation of the PMI.

Postmortem sympathomimetic iris excitability.

BACKGROUND: A reliable estimation of time since death can be important for the law enforcement authorities. The compound method encompassing supravital reactions such as the chemical excitability of the iris can be used to further narrow intervals estimated by temperature-based methods. Postmortem iris excitability was mostly assessed by parasympatholytic or parasympathomimetic substances. Little is known regarding sympathomimetic agents. The present study aims to describe the postmortem iris excitability using the sympathomimetic drug phenylephrine. METHODS: Cadavers were included after body donors gave written informed consent during lifetime. Exclusion criteria were known eye disease, or a postmortem interval exceeding 26 hours. A pupillometer with a minimum measurement range of 0.5 mm was used to determine the horizontal pupil diameter before and 20 minutes after the application of phenylephrine. Increase in pupil diameter was labeled as positive reaction, unchanged pupil diameter was labeled as negative reaction, and decrease in pupil diameter was labeled as paradox reaction. RESULTS: 30 eyes from 16 cadavers (median age = 80.0; 9 males, 7 females) were examined. Initial pupil size was in median 3.5 mm (interquartile range [IQR]: 3.0-4.5 mm) and progressed to 4.0 mm (IQR: 3.5-5.0 mm) 20 minutes after drug instillation. The achieved pupil diameter difference comprised in median 0.5 mm (IQR: 0.0-1.0 mm). A positive reaction was observed in 21 cases. Negative reactions were observed in 5 cases and paradox reactions in 4 cases. Overall, there was a statistically significant difference in diameter between the initial and the reactive pupil (P = 0.0002). CONCLUSION: Although relatively rarely used, sympathomimetic drugs seem to be eligible for chemical postmortem iris excitability. Currently, assessment of postmortem iris excitability usually only involves parasympatholytic and parasympathomimetic agents. The findings of the present study give a hint that the application of a third agent with a sympathomimetic mechanism of action could provide additional information. Further studies assessing such a triple approach in the compound method in comparison with the current gold standard for estimation of time since death are mandatory to ensure reliable results.

Postmortem chondrocyte viability in porcine articular cartilage: Influence of time, temperature, and burial under winter conditions.

The aim of the present study was to investigate the effects of time, temperature, and burial in a natural environment on the viability of chondrocytes in porcine femoral condyles using confocal laser scanning microscopy. Hind trotters from 10 pigs were buried or left unburied. Samples were collected daily and stained with a combination of vital dyes (calcein-AM and ethidium homodimer-1). The chondrocytes showed an intense staining corresponding to their vitality. In the first 3 days, viability decreased slowly and showed no statistical difference between buried and unburied samples. After the first 3 days, it decreased rapidly, with the viability of the buried samples being 66% on day 4, decreasing to 25% on day 8 and to 16% on day 10, while in the unburied samples it decreased to 43% on day 4, 13% on day 8 and 5% on day 10. Our results indicate a time, temperature, and burial dependent decrease in chondrocyte viability and suggest the use of chondrocyte viability as a marker for estimating PMI in both the natural environment and in animals, as well as its potential use in humans.

Post-mortem formation of ethanol: Is 1-propanol a reliable marker? A proof-of-concept study using an in vitro putrefactive environment setup.

Ethanol is the psychoactive substance identified most frequently in post-mortem specimens. Unfortunately, interpreting post-mortem ethanol concentrations can be difficult because of post-mortem alcohol redistribution and the possibility of post-mortem alcohol neogenesis. Indeed, in the time interval between death and sample collection, the decedent may be exposed to non-controlled environments for an extended period, promoting microbial colonization. Many authors report that in the presence of carbohydrates and other biomolecules, various species of bacteria, yeast, and fungi can synthesize ethanol and other volatile substances in vitro and in vivo. The aim of this study was to study the impact of several variables on microbial ethanol production as well as develop a mathematical model that could estimate the microbial-produced ethanol in correlation with the most significant consensual produced higher alcohol, 1-propanol. An experimental setup was developed using human blood samples and cadaveric fragments incubated under strictly anaerobic conditions to produce a novel substrate, "cadaveric putrefactive blood" mimicking post-mortem corpse conditions. The samples were analyzed daily for ethanol and 1-propanol using an HS-GC-FID validated method. The formation of ethanol was evaluated considering different parameters such as putrefactive stage, blood glucose concentration, storage temperature, and storage time. Statistical analysis was performed using the Mann-Whitney non-parametric test and simple linear regression. The results indicate that the early putrefactive stage, high blood glucose concentration, high temperature, and time of incubation increase microbial ethanol production. In addition, the developed mathematical equation confirms the feasibility of using 1-propanol as a marker of post-mortem ethanol production.

Morphometric analysis for determination of larval instars in Dermestes frischii Kugelann and Dermestes undulatus Brahm (Coleoptera: Dermestidae).

Dermestes frischii Kugelann, 1792 and Dermestes undulatus Brahm, 1790 are the most abundant species worldwide at outdoor or indoor crime scenes during the dry and skeletal stages of decomposition. The attribution of larval age in these beetles is problematic due to the variable number of instars, which is influenced by environmental factors. In this study, a morphometric approach was used to look for potential morphological features as evidence of larval stages. Breeding and monitoring were performed for both species in an incubator with a preset temperature of 28°C ± 0.5 without a photoperiod. Morphometric measurements were made on 10 larvae per instar for each species using length, width, and thickness parameters. Linear discriminant analysis was then used to generate decision boundaries that clearly separated larval stages. The cross-validation procedure demonstrated that the morphometric approach successfully discriminated adjacent larval stages in both species with high values of sensitivity and specificity. This less-invasive approach could improve the ability to estimate minPMI in forensic studies of Dermestidae beetles. Future studies may extend this approach to other species and establish good practices for collecting and storing specimens for morphometric analysis.

Ant bite artifacts in a case of hanging.

Artifacts produced by postmortem animal scavenging are commonly encountered at autopsy. Knowledge of the pattern of artifacts produced by postmortem animal scavenging is essential for the correct interpretation of the autopsy finding. In household deaths, such artifacts are usually caused by domestic animals and by small insects such as flies, ants, beetles, etc. Ants are one of the early scavengers which feed on the dead bodies. The artifacts produced by the postmortem ant activity are usually superficial and non-bleeding type. Rarely, in the congested body regions and areas of marked hypostasis, postmortem bleeding artifacts due to ant bites are possible. In the reported case of hanging, such postmortem bleeding artifacts were present over both the legs, predominantly over the left leg. Typical ant bite lesions in the form of superficial excoriations were also present over the peri-ligature area, over and around both nipples and over the lower part of the abdomen. Morphologically, postmortem bleeding artifacts produced by ant bites exhibit four patterns: droplet pattern, stripe pattern, pool pattern, and mixed pattern. In this case, a mixed pattern (droplet pattern and stripe pattern) of postmortem bleeding artifacts was observed. The presence of postmortem bleeding artifacts over the lower limbs was attributed to the pooling of the blood due to suspension of the body, followed by passive escape of blood due to ant bites. The possibility of such artifacts produced by ant bites should be considered when the origin of the lesion is unclear.

Evaluation of porcine decomposition and total body score (TBS) in a central European temperate forest.

The total body score (TBS) is a visual scoring method to scale the succession of decomposition stages. It compares decomposition between cadavers, to connect it with external taphonomic factors and estimate the post-mortem interval. To study decomposition in various climatic environments, pigs are often used as human proxies. Currently, there is one TBS system by Keough et al. (J Forensic Sci. 2017;62:986) for surface-deposited domestic pigs, coming from South Africa. Our study aims to evaluate this method and analyze porcine decomposition in Central Europe to inform forensic research and casework. We conducted an experiment studying six 50 kg pig carcasses in a temperate Swiss forest. Three observers documented decomposition patterns and rated the decomposition stages from photographs based on the porcine TBS model by Keough et al. (J Forensic Sci. 2017;62:986). We documented discrepancies between the carcass decomposition of our specimens and those in the South African study, especially related to the high insect activity in our experiment. Furthermore, we noted factors complicating TBS scoring, including rainfall and scavengers. The agreement between TBS observers from photographs was in the highest agreement category apart from one "substantial agreement" category. Our study is the first in Europe to systematically test the Keough et al. (J Forensic Sci. 2017;62:986) method. The results evidence that regional adaptations are required to be applicable for other environments. We present a modified approach based on experimental observations in a Swiss temperate forest. The identification of regional decomposition patterns and drivers will inform future taphonomy research as well as forensic casework in comparable contexts in Central Europe.