Transient hypoxia drives soil microbial community dynamics and biogeochemistry during human decomposition.

Human decomposition in terrestrial ecosystems is a dynamic process creating localized hot spots of soil microbial activity. Longer-term (beyond a few months) impacts on decomposer microbial communities are poorly characterized and do not typically connect microbial communities to biogeochemistry, limiting our understanding of decomposer communities and their functions. We performed separate year-long human decomposition trials, one starting in spring, another in winter, integrating bacterial and fungal community structure and abundances with soil physicochemistry and biogeochemistry to identify key drivers of microbial community change. In both trials, soil acidification, elevated microbial respiration, and reduced soil oxygen concentrations occurred. Changes in soil oxygen concentrations were the primary driver of microbial succession and nitrogen transformation patterns, while fungal community diversity and abundance was related to soil pH. Relative abundance of facultative anaerobic taxa (Firmicutes and Saccharomycetes) increased during the period of reduced soil oxygen. The magnitude and timing of the decomposition responses were amplified during the spring trial relative to the winter, even when corrected for thermal inputs (accumulated degree days). Further, soil chemical parameters, microbial community structure, and fungal gene abundances remained altered at the end of 1 year, suggesting longer-term impacts on soil ecosystems beyond the initial pulse of decomposition products.

Utilization of qPCR to Determine Duration and Environmental Drivers Contributing to the Persistence of Human DNA in Soil.

Little is known about the underlying mechanisms that contribute to the persistence and degradation of DNA within soil. The goals of this study are to determine the duration of mitochondrial DNA (mtDNA) and nuclear DNA (nuDNA) persistence in soils enriched by surface-level human decomposition and to better understand the contribution of environmental factors. The surface-level decomposition of three human cadavers was documented over 11 weeks. Based on quantitative PCR results, we found nuDNA to persist in soils six weeks post-placement, while mtDNA was recoverable for the entire 11-week decomposition period. Principle components analyses and Spearman's rank correlations revealed that (1) time, (2) total body score, and (3) weekly average air temperature were significantly correlated with concentrations of nuDNA and mtDNA in soil, suggesting these factors play a role in the degradation of DNA in soils.

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.

Sr-Pb isotope differences in pre- and post-burial human bone, teeth, and hair keratin: implications for isotope forensics.

The isotopic signatures of human tissues can provide valuable information on geographic origin for medicolegal investigations involving unidentified persons. It is important to understand the impact of diagenetic processes on isotopic signatures, as alterations could result in incorrect estimation of geographic origin. This study examines alterations in isotope signatures of different tissues of five human body donors studied throughout decomposition at the Forensic Anthropology Research Facility (FARF), San Marcos, TX. Two body donors were buried, two were placed in open pits, and one was first allowed to naturally mummify and then buried. Remains were recovered after a period of 7-34 months. The preplacement and post-recovery Sr-Pb isotope data of scalp hair, bone (iliac and tibia), and tooth enamel and dentine were compared. The hair samples record significant shifts in Sr-Pb isotope compositions, with hair keratin Pb isotope composition shifting towards the Pb signature of local soil samples. Hair keratin Sr isotope compositions were altered by the burial environment and possibly also by the lab sample cleaning method. The spongy iliac bone samples show inconsistencies in the recoverability of the preplacement Sr-Pb isotope signatures. The post-placement signatures of the buried donors show slight elevation over preplacement signatures. The post-placement signatures of donors placed in open pits are significantly elevated. The tibia and dental samples record the most consistent isotopic data with the least alteration. These more densely mineralised elements show good recoverability of the preplacement isotope signatures in burials and open pits and are thus deemed better targets for forensic investigative purposes.

ICPUTRD: Image Cloud Platform for use in tagging and research on decomposition.

Human decomposition studies aim to understand the various factors influencing human decay to assess the deceased and develop postmortem interval (PMI) estimation methods. These types of studies are typically conducted through physical experiments examining the deceased; however, big data systems have the potential to transform how large-scale forensic anthropology research questions can be addressed with curated images of donors with known demographic, climatic, and postmortem historical data. This study introduces ICPUTRD (Image Cloud Platform for Use in Tagging and Research on Decomposition), a web-based software system, which enables forensic scientists to easily access, enhance (or curate), and analyze very large photographic collections documenting the longitudinal process of human decomposition. ICPUTRD, a JavaScript-based application, was designed and built through a combination of the Waterfall and Agile software development life-cycle methods and provides an image search and tagging features with a predefined nomenclature of forensic-related keywords. To evaluate the system, a user study was conducted, involving 27 participants who completed pre- and post-study surveys and three research tasks. Analysis of the study results confirmed the feasibility and practicality of ICPUTRD to facilitate aspects of forensic research and casework involving large collections of digital photographs of human decomposition. It was observed that the nomenclature lacked certain law enforcement keywords, so future work will focus on expanding it to ensure ICPUTRD is suited for all its intended users.

Retrospective analysis of factors affecting rate of skeletonization within a tropical climate.

Estimating the post mortem interval (PMI) in skeletonized cases is an extremely challenging exercise. Sri Lanka lacks adequate taphonomic research which is a serious limitation when assessing PMI in forensic death investigations. Methods that have been proposed to estimate PMI using the total body score (TBS) and accumulated degree days (ADD) are mainly based on data from continental and temperate climates and have shown less reliability in tropical climates. With the intention of developing a region-specific, evidence-based guide which would be applicable to tropical climates like Sri Lanka, we selected thirteen skeletonized remains with known PMIs from forensic case records and analysed their taphonomy in relation to selected weather data. We also compared the ADD values within our dataset with reference ranges calculated using published formula. All except one were found from outdoor locations. The TBS ranged from 24 to 32 and had a weak positive correlation with the PMI. The earliest appearance of skeletonization was 15 days in a body found indoors. The highest rate of skeletonization was seen in a body with a TBS of 32 and a PMI of 23 days. The average daily temperature and relative humidity were similar across all the cases however, the amount of rainfall varied. Bodies exposed to monsoon rains (n = 6) had a lower mean rate of skeletonization compared to those that were not exposed (n = 4) suggesting lower rates of decomposition during periods of heavy rainfall. No correlation was found between ADD and TBS. In 9 (69.2%) cases, the actual ADD was much lower than reference ADD ranges for TBS values, indicating poor applicability of TBS and ADD based formulae in estimating PMI within the Sri Lankan climate. Our study shows a strong need for taphonomic and entomological research in tropical climates to further explore the impact of monsoons on biotic and abiotic factors affecting skeletonization.

Estimating the time of human decomposition based on skeletal muscle biopsy samples utilizing an untargeted LC-MS/MS-based proteomics approach.

Accurate estimation of the postmortem interval (PMI) is crucial in forensic medico-legal investigations to understand case circumstances (e.g. narrowing down list of missing persons or include/exclude suspects). Due to the complex decomposition chemistry, estimation of PMI remains challenging and currently often relies on the subjective visual assessment of gross morphological/taphonomic changes of a body during decomposition or entomological data. The aim of the current study was to investigate the human decomposition process up to 3 months after death and propose novel time-dependent biomarkers (peptide ratios) for the estimation of decomposition time. An untargeted liquid chromatography tandem mass spectrometry-based bottom-up proteomics workflow (ion mobility separated) was utilized to analyse skeletal muscle, collected repeatedly from nine body donors decomposing in an open eucalypt woodland environment in Australia. Additionally, general analytical considerations for large-scale proteomics studies for PMI determination are raised and discussed. Multiple peptide ratios (human origin) were successfully proposed (subgroups < 200 accumulated degree days (ADD), < 655 ADD and < 1535 ADD) as a first step towards generalised, objective biochemical estimation of decomposition time. Furthermore, peptide ratios for donor-specific intrinsic factors (sex and body mass) were found. Search of peptide data against a bacterial database did not yield any results most likely due to the low abundance of bacterial proteins within the collected human biopsy samples. For comprehensive time-dependent modelling, increased donor number would be necessary along with targeted confirmation of proposed peptides. Overall, the presented results provide valuable information that aid in the understanding and estimation of the human decomposition processes.

Decomposition and insect succession on human cadavers in a humid, continental (Dfb) climate (Quebec, Canada).

Forensic entomology has been developing globally for decades. Despite this discipline being used in criminal investigations around the world, only a few controlled studies have been performed on human cadavers in human decomposition facilities, with the majority of these being conducted in warm and often dry climates. Therefore, the purpose of our research was to catalogue the first published data on insects associated with decomposed human bodies in a humid, continental (Dfb) climate. Specifically, our objective was to document the diversity and succession of the entomofauna associated with human cadavers throughout the decomposition process, in the Quebec province of Canada, during the summer season. Two human cadavers were studied in 2020 at the site for Research in Experimental and Social Thanatology, REST, located in Bécancour (Quebec, Canada). Insects (and other arthropods) were regularly sampled by visual observations, collection from the cadavers, and by using an entomological net and pitfall traps. Our results highlight that the decomposition process is a heterogeneous and complex process in Quebec, with cadavers showing signs of precocious desiccation/mummification. In addition, our observations confirm that the presence of superficial skin lesions accelerates the colonization of blow flies (Diptera: Calliphoridae) and, consequently, the process of decomposition. Finally, we were able to discriminate between "early colonizers" (e.g., Calliphoridae Lucilia sp. or Calliphora livida), "late colonizers" (e.g., larvae of Piophilidae or Heleomyzidae), and "non-specific colonizers." We also officially report the first observation of Cochliomyia macellaria (Diptera: Calliphoridae) in Quebec. These findings will provide new information to help medico-legal death investigations by determining the minimum time elapsed since death and the circumstances surrounding death.

Complexity of human death: its physiological, transcriptomic, and microbiological implications.

Human death is a complex, time-governed phenomenon that leads to the irreversible cessation of all bodily functions. Recent molecular and genetic studies have revealed remarkable experimental evidence of genetically programmed cellular death characterized by several physiological processes; however, the basic physiological function that occurs during the immediate postmortem period remains inadequately described. There is a paucity of knowledge connecting necrotic pathologies occurring in human organ tissues to complete functional loss of the human organism. Cells, tissues, organs, and organ systems show a range of differential resilience and endurance responses that occur during organismal death. Intriguingly, a persistent ambiguity in the study of postmortem physiological systems is the determination of the trajectory of a complex multicellular human body, far from life-sustaining homeostasis, following the gradual or sudden expiry of its regulatory systems. Recent groundbreaking investigations have resulted in a paradigm shift in understanding the cell biology and physiology of death. Two significant findings are that (i) most cells in the human body are microbial, and (ii) microbial cell abundance significantly increases after death. By addressing the physiological as well as the microbiological aspects of death, future investigations are poised to reveal innovative insights into the enigmatic biological activities associated with death and human decomposition. Understanding the elaborate crosstalk of abiotic and biotic factors in the context of death has implications for scientific discoveries important to informing translational knowledge regarding the transition from living to the non-living. There are important and practical needs for a transformative reestablishment of accepted models of biological death (i.e., artificial intelligence, AI) for more precise determinations of when the regulatory mechanisms for homeostasis of a living individual have ceased. In this review, we summarize mechanisms of physiological, genetic, and microbiological processes that define the biological changes and pathways associated with human organismal death and decomposition.

Body Mass Index (BMI) Impacts Soil Chemical and Microbial Response to Human Decomposition.

Microorganisms are key decomposers of vertebrate mortalities, breaking down body tissues and impacting decomposition progress. During human decomposition, both extrinsic environmental factors and intrinsic cadaver-related factors have the potential to impact microbial decomposers either directly or indirectly via altered physical or chemical conditions. While extrinsic factors (e.g., temperature, humidity) explain some variation in microbial response during human decomposition in terrestrial settings, recent work has noted that even under the same environmental conditions, individuals can have different decomposition patterns, highlighting the potential for intrinsic factors to impact microbial decomposers. The goal of this study was to investigate the effects of several intrinsic factors (age, sex, diseases at time of death, and body mass index [BMI]) on chemical and microbial changes in decomposition-impacted soils. In a field study conducted at the University of Tennessee Anthropology Research Facility, soils were collected from the decomposition-impacted area surrounding 19 deceased human individuals through the end of active decomposition. Soil physicochemical parameters were measured, and microbial (bacterial and fungal) communities were assessed via amplicon sequencing. BMI was shown to explain some variation in soil pH and microbial response to human decomposition. Hierarchical linear mixed (HLM) effects models revealed that BMI category significantly explained variation in pH response within decomposition-impacted soils over time (HLM F = 9.647; P < 0.001). Additionally, the relative abundance of soil Saccharomycetes in decomposition soils under underweight donors displayed little to no changes (mean maximum change in relative abundance, +6.6%), while all other BMI categories displayed an increased relative abundance of these organisms over time (normal, +50.6%; overweight, +64.4%; and obese, +64.6%) (HLM F = 3.441; P = 0.11). Together, these results reveal intrinsic factors influencing decomposition patterns, especially within the soil environment, and suggest BMI is an important factor for controlling decomposition processes. IMPORTANCE This work begins to address questions about interindividual variation in vertebrate decomposition attributed to intrinsic factors, that is, properties of the carcass or cadaver itself. Most research on factors affecting decomposition has focused on the extrinsic environment, such as temperature or humidity. While these extrinsic factors do explain some variation in decomposition patterns, interindividual variability is still observed. Understanding how intrinsic factors influence microbial decomposers will help reveal the ecological impacts of decomposition. This work also has forensic applications, as soil chemical and biological changes have been suggested as indicators of postmortem interval. We reveal factors that explain variation in the decomposition environment that should be considered in these estimates. This is particularly important as we consider the implications of variations in human populations due to diet, age, BMI, disease, toxicological loading, etc. on forensic investigations dealing with decomposing remains.

Taphonomic study on drowned victims in a non-sequestered aquatic environment in the Mediterranean Sea.

Human decomposition in sea water poses several challenges to forensic practitioners tasked with the analysis of drowned bodies. Postmortem changes in the marine environment have not been extensively investigated and the current literature is mainly based on bodies retrieved from shallow waters or on limited samples. On 18 April 2015, a fishing boat carrying allegedly 1,000 migrants sank in the Mediterranean Sea. In a fifteen-month time span, humanitarian missions were carried out to recover the bodies from the sea. The present study investigates postmortem changes on the drowned victims in a non-sequestered environment in the Mediterranean Sea. A retrospective study was performed by two observers on the autopsy photographic records in the series of bodies recovered from the open sea. For 184 bodies, the postmortem changes were evaluated according to facial, body, limb, and total aquatic decomposition score (FADS, BADS, LADS, TADS, respectively). Furthermore, a modification to the current scoring system that divides upper and lower limbs was suggested. The interobserver agreement was assessed using Krippendorff's alpha coefficient. Possible relations between the decomposition scores and PMSI were investigated with Pearson correlation analysis. According to the sequence of the recovery missions, increasing FADS, BADS, LADS, and TADS were observed. The modified scoring system showed a strong agreement between observers, allowing a more accurate description of the actual extent of decomposition. The scores showed a significant relation with the PMSI (p < 0.01). Prolonged submersion in the open marine environment was confirmed to show increasing decomposition rates, from moderate decay to full disarticulation. This study provides a descriptive unicum of the postmortem changes in the open sea, which may contribute to strengthen the discipline and aid description of bodies recovered in similar circumstances, especially if a body needs to be associated to a disaster or period with respect to another, thus facilitating families or authorities in the search for specific victims.

Characterization of Bacterial Community Dynamics of the Human Mouth Throughout Decomposition via Metagenomic, Metatranscriptomic, and Culturing Techniques.

The postmortem microbiome has recently moved to the forefront of forensic research, and many studies have focused on the idea that predictable fluctuations in decomposer communities could be used as a "microbial clock" to determine time of death. Commonly, the oral microbiome has been evaluated using 16S rRNA gene sequencing to assess the changes in community composition throughout decomposition. We sampled the hard palates of three human donors over time to identify the prominent members of the microbiome. This study combined 16S rRNA sequencing with whole metagenomic (MetaG) and metatranscriptomic (MetaT) sequencing and culturing methodologies in an attempt to broaden current knowledge about how these postmortem microbiota change and might function throughout decomposition. In all four methods, Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes were the dominant phyla, but their distributions were insufficient in separating samples based on decomposition stage or time or by donor. Better resolution was observed at the level of genus, with fresher samples from decomposition clustering away from others via principal components analysis (PCA) of the sequencing data. Key genera in driving these trends included Rothia; Lysinibacillus, Lactobacillus, Staphylococcus, and other Firmicutes; and yeasts including Candida and Yarrowia. The majority of cultures (89%) matched to sequences obtained from at least one of the sequencing methods, while 11 cultures were found in the same samples using all three methods. These included Acinetobacter gerneri, Comamonas terrigena, Morganella morganii, Proteus vulgaris, Pseudomonas koreensis, Pseudomonas moraviensis, Raoutella terrigena, Stenotrophomonas maltophilia, Bacillus cereus, Kurthia zopfii, and Lactobacillus paracasei. MetaG and MetaT data also revealed many novel insects as likely visitors to the donors in this study, opening the door to investigating them as potential vectors of microorganisms during decomposition. The presence of cultures at specific time points in decomposition, including samples for which we have MetaT data, will yield future studies tying specific taxa to metabolic pathways involved in decomposition. Overall, we have shown that our 16S rRNA sequencing results from the human hard palate are consistent with other studies and have expanded on the range of taxa shown to be associated with human decomposition, including eukaryotes, based on additional sequencing technologies.

Patterns of striped skunk scavenging on human remains.

Scavenging is a known behavior of striped skunks (Mephitis mephitis), though it is rarely documented. Striped skunks were observed and documented scavenging nine human donors at the Forensic Investigation Research Station (FIRS) in Whitewater, Colorado from October 2017 through September 2019. Human remains are placed on the surface, outdoors, in a fenced area, and monitored daily through notes, photographs, and multiple systems for scoring decomposition. Motion-activated game cameras are strategically positioned to monitor potential scavenger activity within the facility. When scavenging occurred, game camera images were temporally correlated to daily photographs and notes to confirm the source of observed defects. Striped skunks at the FIRS scavenged the soft tissue of the arms, legs, genitals, and torso. Game camera images documented skunks digging, climbing on top of remains, repositioning limbs, and using their paws to manipulate tissue. These activities impacted soft tissue, bone, and the surrounding environment. Scavenging patterns varied seasonally, with higher frequency and lower selectivity in the winter. Striped skunks are common throughout most of North America and are likely to scavenge human remains when available. Understanding patterns and behaviors of specific scavengers can help investigators accurately interpret defects found on and around human remains.

Human Bone Proteomes before and after Decomposition: Investigating the Effects of Biological Variation and Taphonomic Alteration on Bone Protein Profiles and the Implications for Forensic Proteomics.

Bone proteomic studies using animal proxies and skeletonized human remains have delivered encouraging results in the search for potential biomarkers for precise and accurate post-mortem interval (PMI) and the age-at-death (AAD) estimation in medico-legal investigations. The development of forensic proteomics for PMI and AAD estimation is in critical need of research on human remains throughout decomposition, as currently the effects of both inter-individual biological differences and taphonomic alteration on the survival of human bone protein profiles are unclear. This study investigated the human bone proteome in four human body donors studied throughout decomposition outdoors. The effects of ageing phenomena (in vivo and post-mortem) and intrinsic and extrinsic variables on the variety and abundancy of the bone proteome were assessed. Results indicate that taphonomic and biological variables play a significant role in the survival of proteins in bone. Our findings suggest that inter-individual and inter-skeletal differences in bone mineral density (BMD) are important variables affecting the survival of proteins. Specific proteins survive better within the mineral matrix due to their mineral-binding properties. The mineral matrix likely also protects these proteins by restricting the movement of decomposer microbes. New potential biomarkers for PMI estimation and AAD estimation were identified. Future development of forensic bone proteomics should include standard measurement of BMD and target a combination of different biomarkers.

Perspectives on the establishment of a canadian human taphonomic facility: The experience of REST[ES].

REST[ES] is the first Canadian human taphonomic facility (HTF) dedicated to research and training relating to human decomposition in a northern temperate climate. The following paper outlines the measures taken to successfully establish, open and operate this novel Canadian HTF with particular focus on: project team and partnerships, facility location, approvals and permits, infrastructure and social acceptability. It is intended that our experience of establishing REST[ES] may serve as an example to help others with the establishment of future HTFs, thus contributing to the expansion in the global accessibility to human decomposition research and training.

Plants to Remotely Detect Human Decomposition?

In the USA, 100 000 people go missing every year. Difficulty in the rapid identification of sites of human decomposition complicates the recovery of bodies, especially in forests. We propose that spectral responses in tree and shrub canopies could act as guides to find cadavers using remote sensing platforms for societal benefit.

Comparative Decomposition of Humans and Pigs: Soil Biogeochemistry, Microbial Activity and Metabolomic Profiles.

Vertebrate decomposition processes have important ecological implications and, in the case of human decomposition, forensic applications. Animals, especially domestic pigs (Sus scrofa), are frequently used as human analogs in forensic decomposition studies. However, recent research shows that humans and pigs do not necessarily decompose in the same manner, with differences in decomposition rates, patterns, and scavenging. The objective of our study was to extend these observations and determine if human and pig decomposition in terrestrial settings have different local impacts on soil biogeochemistry and microbial activity. In two seasonal trials (summer and winter), we simultaneously placed replicate human donors and pig carcasses on the soil surface and allowed them to decompose. In both human and pig decomposition-impacted soils, we observed elevated microbial respiration, protease activity, and ammonium, indicative of enhanced microbial ammonification and limited nitrification in soil during soft tissue decomposition. Soil respiration was comparable between summer and winter, indicating similar microbial activity; however, the magnitude of the pulse of decomposition products was greater in the summer. Using untargeted metabolomics and lipidomics approaches, we identified 38 metabolites and 54 lipids that were elevated in both human and pig decomposition-impacted soils. The most frequently detected metabolites were anthranilate, creatine, 5-hydroxyindoleacetic acid, taurine, xanthine, N-acetylglutamine, acetyllysine, and sedoheptulose 1/7-phosphate; the most frequently detected lipids were phosphatidylethanolamine and monogalactosyldiacylglycerol. Decomposition soils were also significantly enriched in metabolites belonging to amino acid metabolic pathways and the TCA cycle. Comparing humans and pigs, we noted several differences in soil biogeochemical responses. Soils under humans decreased in pH as decomposition progressed, while under pigs, soil pH increased. Additionally, under pigs we observed significantly higher ammonium and protease activities compared to humans. We identified several metabolites that were elevated in human decomposition soil compared to pig decomposition soil, including 2-oxo-4-methylthiobutanoate, sn-glycerol 3-phosphate, and tryptophan, suggesting different decomposition chemistries and timing between the two species. Together, our work shows that human and pig decomposition differ in terms of their impacts on soil biogeochemistry and microbial decomposer activities, adding to our understanding of decomposition ecology and informing the use of non-human models in forensic research.

Death is in the air: Confirmation of decomposition without a corpse.

This case report summarises the investigation of a death scene in the trunk of a car. Air sampling, laser-induced breakdown spectroscopy, and gas chromatography/mass spectrometry on samples of carpet and tyre well scrapings from the vehicle's trunk were utilised to confirm the presence of a human decompositional event even though no human remains were discovered in the vehicle. Air sampling has been used in numerous industries for many decades, but only recently has been applied to forensic investigations although it has been at the centre of controversy over the use of this technique in such cases. This report also describes the value of such investigative tools and points to the discovery of evidence, which, without the use of these techniques, would not have been identified.

An interdisciplinary review of the thanatomicrobiome in human decomposition.

Death does not occur instantaneously and organs do not decompose at the same rate or in the same way. Nulligravid human uteri and prostate glands are the last internal organs to deteriorate during decomposition; however, the reason for this very important observation is still enigmatic. Recent studies have elucidated that the composition and abundance of microbes in the human thanatomicrobiome (microbiome of death) varies by organ and changes as a function of time and temperature. The ileocecal area has the largest absolute postmortem burden that spreads to the liver and spleen and continues to the heart and brain depending on the cause of death. To truly understand the mechanisms of microbial assembly during decomposition, a thorough examination of different strategies utilized by the trillions of microbes that colonize decaying tissues is needed from a multi-organ and multidisciplinary approach. In this review, we highlight interdisciplinary research and provide an overview of human decomposition investigations of thanatomicrobiomic changes in internal organs.

Monitoring the extent of vertical and lateral movement of human decomposition products through sediment using cholesterol as a biomarker.

Due to the lack of human decomposition research facilities available in different geographical regions, the extent of movement of human decomposition products from a cadaver into various sedimentary environments, in different climates, has not been able to be studied in detail. In our study, a human cadaver was placed on the surface of a designated plot at the Australian Facility for Taphonomic Experimental Research (AFTER), the only human decomposition facility in Australia, where the natural process of decomposition was allowed to progress over 14days in the Australian summer. Sediment columns (approximately 1m deep) were collected at lateral distances of 0.25m, 0.5m, 1.0m and 2.5m in each of four directions from the centre of the torso. Plot elevation and weather data were also collected. Each sediment column was subdivided, dried and homogenised. A sample was isolated from each sediment subdivision, extracted with hexane, and the hexane extract cleaned with citrate buffer (pH 3), filtered and spiked with cholesterol-D7 internal standard. After derivatisation with BSTFA+1% TMCS, cholesterol was monitored in the samples using targeted gas chromatography tandem mass spectrometry analysis. A positive result for decomposition products was given if the cholesterol abundance in the test sample was higher than that detected in the 'control' samples of a similar substrate type collected prior to cadaver placement. Within the confines of the experimental design and the measured parameters, lateral leaching was observed over distances of up to 2.5m from the centre of the torso, which was the maximum distance tested in the study. Vertical leaching was detected to depths of up to 49cm below the ground surface. Such data can aid the development of policies related to plot sizing and sediment renewal and regeneration at other human decomposition facilities and at cemeteries. The density and distribution of cholesterol surrounding the cadaver in this study can also help forensic investigators interpret cases involving remains that have been moved or scavenged.