Biochemistry Changes That Occur after Death: Potential Markers for Determining Post-Mortem Interval

Death is likely to result in very extensive biochemical changes in all body tissues due to lack of circulating oxygen, altered enzymatic reactions, cellular degradation, and cessation of anabolic production of metabolites. These biochemical changes may provide chemical markers for helping to more accurately determine the time since death (post-mortem interval), which is challenging to establish with current observation-based methodologies. In this study blood pH and changes in concentration of six metabolites (lactic acid, hypoxanthine, uric acid, ammonia, NADH and formic acid) were examined post-mortem over a 96 hour period in blood taken from animal corpses (rat and pig) and blood from rats and humans stored in vitro. The pH and the concentration of all six metabolites changed post-mortem but the extent and rate of change varied. Blood pH in corpses fell from 7.4 to 5.1. Concentrations of hypoxanthine, ammonia, NADH and formic acid all increased with time and these metabolites may be potential markers for post-mortem interval. The concentration of lactate increased and then remained at an elevated level and changes in the concentration were different in the rat compared to the human and pig. This is the first systematic study of multiple metabolic changes post-mortem and demonstrates the nature and extent of the changes that occur, in addition to identifying potential markers for estimating post-mortem interval.     

常绿阔叶林片段中木荷凋落叶分解速率及中小型土壤节肢动物群落的结构动态

2004年6月-2006年4月,选择浙、闽、赣交界山地的4个森林片段作为研究地点,并以浙西的古田山国家级自然保护区作为连续森林对照,采用尼龙网袋法,研究了不同常绿阔叶林群落优势种木荷凋落叶的分解速率及土壤节肢动物群落的结构和动态.结果表明：5个研究地共获土壤节肢动物1050个,分属8纲23目,其中鳞翅目、膜翅目、弹尾目和双翅目占个体总数的10%以上,为该地区的优势类群.片段化对群落物种组成的影响主要体现在稀有类群的差异上.不同分解阶段各种土壤节肢动物类群优势度因其功能的不同而各异.经过2年的分解,各研究地木荷凋落叶损失量为60%～70%,研究地凋落物中土壤节肢动物的各项多样性指标均有一定的变化,且与连续森林的变化规律不一致.     

Sarcomeric disorganization in post-mortem fish muscles.

1. The post-mortem evolution of protein pattern in fish striated muscle was followed by SDS-PAGE, after different conditions of storage time and temperature. 2. Sarcoplasmic and sarcomeric fractions were analyzed respectively by low and high ionic strength extractions of fish muscle samples. 3. No evident modification of electrophoretic patterns was observed during the pre-rigor mortis period. 4. The high mol. wt proteins titin and nebulin were highly sensitive to proteolysis during the rigor mortis period. 5. Myosin extraction was predominantly influenced by the storage temperature. The myosin content of the extracts decreased during the rigor mortis period at storage temperatures greater than 8 degrees C. 6. alpha-Actinin was very resistant to proteolysis, but could be released from Z-disc structure during post-mortem aging.     

Refining decomposition stage estimation: A study of seasonal insect composition and carcass weight change

In this study, we explore the interplay of seasonal insect assemblages and carcass weight change to refine the estimation of minimum post-mortem interval (PMI_min), a key variable in forensic investigations. By integrating these parameters, we shed light on their combined influence on decomposition stages, thus paving the way for a more robust method of PMI_min estimation. Our approach relies on rigorous field studies examining pig carcasses across spring, summer and autumn, with comprehensive measurements of carcass weight and detailed documentation of insect succession. We observed distinct insect assemblages and weight-change trends characteristic of each season, underlining the influence of seasonal variability on decomposition dynamics. To further enhance the reliability of our model, we combined the novel parameter of carcass weight change with the conventional total body score (TBS) method, ensuring a more objective assessment of decomposition stages. Although our study indicates promising advancements, it also acknowledges the limitations of using a single pig carcass per season, suggesting future research should incorporate larger sample sizes.     

Predicting postmortem interval based on microbial community sequences and machine learning algorithms

Microbes play an essential role in the decomposition process but were poorly understood in their succession and behaviour. Previous researches have shown that microbes show predictable behaviour that starts at death and changes during the decomposition process. Research of such behaviour enhances the understanding of decomposition and benefits estimating the postmortem interval (PMI) in forensic investigations, which is critical but faces multiple challenges. In this study, we combined microbial community characterization, microbiome sequencing from different organs (i.e. brain, heart and cecum) and machine learning algorithms [random forest (RF), support vector machine (SVM) and artificial neural network (ANN)] to investigate microbial succession pattern during corpse decomposition and estimate PMI in a mouse corpse system. Microbial communities exhibited significant differences between the death point and advanced decay stages. Enterococcus faecalis, Anaerosalibacter bizertensis, Lactobacillus reuteri, and so forth were identified as the most informative species in the decomposition process. Furthermore, the ANN model combined with the postmortem microbial data set from the cecum, which was the best combination among all candidates, yielded a mean absolute error of 1.5 ± 0.8 h within 24-h decomposition and 14.5 ± 4.4 h within 15-day decomposition. This integrated model can serve as a reliable and accurate technology in PMI estimation.     

Microbial traces and their role in forensic science

Forensic microbiology, also known as the microbiology of death, is an emerging branch of science that is still underused in criminal investigations. Some of the cases might be difficult to solve with commonly used forensic methods, and then they become an operational field for microbiological and mycological analyses. The aim of our review is to present significant achievements of selected studies on the thanatomicrobiome (micro-organisms found in the body, organs and fluids after death) and epinecrotic community (micro-organisms found on decaying corpses) that can be used in forensic sciences. Research carried out as a part of the forensic microbiology deals with the thanatomicrobiome and the necrobiome—communities of micro-organisms that live inside and outside of a putrefying corpse. Change of species composition observed in each community is a valuable feature that gives a lot of information related to the crime. It is mainly used in the estimation of post-mortem interval (PMI). In some criminal investigations, such noticeable changes in the microbiome and mycobiome can determine the cause or the actual place of death. The microbial traces found at the crime scene can also provide clear evidence of guilt. Nowadays, identification of micro-organisms isolated from the body or environment is based on metagenome analysis and 16S rRNA gene amplicon-based sequencing for bacteria and ITS rRNA gene amplicon-based sequencing for fungi. Cultivation methods are still in use and seem to be more accurate; however, they require much more time to achieve a final result, which is an unwanted feature in any criminal investigation.     

Temporal dynamics of biotic and abiotic drivers of litter decomposition

Climate, litter quality and decomposers drive litter decomposition. However, little is known about whether their relative contribution changes at different decomposition stages. To fill this gap, we evaluated the relative importance of leaf litter polyphenols, decomposer communities and soil moisture for litter C and N loss at different stages throughout the decomposition process. Although both microbial and nematode communities regulated litter C and N loss in the early decomposition stages, soil moisture and legacy effects of initial differences in litter quality played a major role in the late stages of the process. Our results provide strong evidence for substantial shifts in how biotic and abiotic factors control litter C and N dynamics during decomposition. Taking into account such temporal dynamics will increase the predictive power of decomposition models that are currently limited by a single‐pool approach applying control variables uniformly to the entire decay process.     

RNA 检测技术在法医病理学中的应用研究

DNA技术广泛应用于法医学中,RNA分析技术为调查疾病和死亡原因提供依据,RNA检测可以作为法医病理学检测一个有效的工具。RNA还可以应用在损伤时间和死亡后间隔时间的检测。在法医案件分析中,体液中细胞特异mRNA的表达的分子检测已经为DNA分析的重要补充手段。本文综述了RNA检测技术在法医病理学中的研究进展及其应用。     

"Muscle to meat" molecular events and technological transformations: the proteomics insight

Cellular death is characterized by a complex pattern of molecular events that depend on cell type. Specifically, muscle cells first undergo rigor mortis due to ATP depletion, and later, on the time scale of days, muscle fiber degradation due to proteolytic enzyme activity. In the present review, we will refer to proteomic investigations on the post-mortem evolution of the protein patterns of animal muscle cells. These studies, carried out with the application of either bottom-up or top-down methods, are relevant for understanding the biochemical reactions that i) convert muscle to meat, ii) are associated with meat aging and iii) impact on meat tenderness, a feature of significant commercial value. We also report on the proteomic investigations that have been made to analyze the transformation of meat in industrial processes. These studies are primarily aimed at identifying protein patterns and/or individual proteins diagnostic of the quality of the final product.     

Identification of brain cell death associated proteins in human post-mortem cerebrospinal fluid

Following any form of brain insult, proteins are released from damaged tissues into the cerebrospinal fluid (CSF). This body fluid is therefore an ideal sample to use in the search for biomarkers of neurodegenerative disorders and brain damage. In this study, we used human post-mortem CSF as a model of massive brain injury and cell death for the identification of such protein markers. Pooled post-mortem CSF samples were analyzed using a protocol that combined immunoaffinity depletion of abundant CSF proteins, off-gel electrophoresis, SDS-PAGE and protein identification by LC-MS/MS. A total of 299 proteins were identified, of which 172 proteins were not previously described to be present in CSF. Of these 172 proteins, more than 75% have been described as intracellular proteins suggesting that they were released from damaged cells. Immunoblots of a number of proteins were performed on individual post-mortem CSF samples and confirmed elevated concentrations in post-mortem CSF compared to ante-mortem CSF. Interestingly, among the proteins specifically identified in the post-mortem CSF, several have been previously described as biochemical markers of brain damage.     

Elephant dung decomposition and coprophilous fungi in two habitats of semi-arid Botswana

In order to understand the impact of habitat changes on ecosystem processes caused by increased populations of elephants, elephant dung decomposition was studied in semi-arid Botswana. Dung decomposition rates were studied with and without the presence of arthropods, using pairs of exposed dung and dung enclosed in nylon-mesh bags, respectively. Dung decomposition rates were lower in the absence of arthropods. The rates in the late wet season were higher in the scrubland than in the woodland. In the early dry season, immediately after the wet season, the rates were higher in the woodland than in the scrubland. The difference in decomposition rates between habitats was attributed to microclimatic conditions created by vegetation cover. With regard to fungal succession, Cladosporium cladosporioides and Eurotium brefeldianum occurred only in the late stages of dung decomposition whereas Talaromyces helicus, Cercophora coprophila and Sporormiella minima occurred in all the stages. Although there was no significant difference in Shannon–Weiner fungal species diversity index between habitats, seasons, dung ages and laboratory incubation periods, there were significant differences in fungal community composition between these parameters. Species richness was higher in the late wet season than in the early dry season, indicating the importance of moist conditions for a large diversity of fungal species.     

Entomofauna resource distribution associated with pig cadavers in Bogotá DC

A cadaver represents a temporal energy‐loaded resource, which provides arthropods with food, protection and a place in which to find a mate. Insects are usually the first organisms to discover and colonize a cadaver; as decomposition progresses, insects colonize cadavers in a predictable sequence. This work aimed to establish cadaverous entomofauna relationships with regard to stages of decomposition and environmental conditions using multiple correspondence analysis and thereby to identify the way in which insects distribute a perishable and changing resource. Entomofauna were thus collected in a semi‐rural area near Bogotá from the cadavers of three pigs (Sus scrofa L.) which had been shot. Environmental variables were recorded for each sampling. Multiple correspondence analyses were carried out for adult forms belonging to Diptera and Coleoptera families and stages of decomposition, and for Diptera and Coleoptera adult forms and environmental conditions. Stages of decomposition were a primary determining factor for structuring four guilds of entomofauna. However, environmental conditions influenced insect activity and were therefore a relevant factor in the structure of the entomofauna community. The results showed that the insects' distribution of available resources was related to changes in the stage of decomposition.     

Effect of body length and temperature on the crawling speed of Protophormia terraenovae larvae (Robineau-Desvoidy) (Diptera Calliphoridae)

This work investigates variation in the crawling speed of Protophormia terraenovae larvae (Robineau-Desvoidy) (Diptera Calliphoridae) as a function of body length and ambient temperature. A video-tracking system was used to follow the movement of larvae in an experimental arena, and to estimate their average crawling speed. Recordings were carried out at various temperatures using larvae of different developmental stages. As temperature increased, the larvae crawled at a faster speed. Furthermore, speed increased as a function of body length. We present a general allometric relationship to express the average crawling speed of larvae as a combined function of temperature and body length, expressed in the following equation: speed (cm/min)=5.45 x log[length (mm)]+0.66 x temperature (degrees C)-12.8. These results are particularly useful in forensic entomology for estimating the time that elapsed following the larva's departure from the corpse. This information can provide more accuracy for post-mortem interval estimations and a better comprehension of larvae behaviour and displacement.     

氨基酸硫酸镧固体配合物热分解过程研究

报道了用ＴＧ—ＤＴＧ法研究七个氨基酸硫酸镧固体配合物的热分解过程。研究表明,配合物有一系列分解阶段,多数均经生成低水合物、无水盐等过程。最终分解产物均为氧化镧。     

Larval growth rates of the blowfly, Calliphora vicina, over a range of temperatures

Blowfly larvae (Diptera: Calliphoridae) fulfil an important ecological function in the decomposition of animal remains. They are also used extensively in forensic entomology, predominantly to establish a minimum time since death, or a minimum post-mortem interval, using the larval length as a 'biological clock'. This study examined the larval growth rate of a forensically important fly species, Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae) at temperatures of between 4 degrees C and 30 degrees C, under controlled laboratory conditions. The laboratory flies had been trapped initially in London, U.K. The minimum developmental temperature was estimated to be 1 degrees C and 4700 accumulated degree hours (ADH) were required for development from egg hatch to the point of pupariation. Lines fitted to the laboratory larval growth data were found to adequately explain the growth of larvae in the field. The nature of variation in growth rates from geographically isolated populations is discussed.     

Phylogenetic relationships of annelids,molluscs, and arthropods evidenced from molecules and morphology

Annelids and arthropods have long been considered each other's closest relatives, as evidenced by similarities in their segmented body plans. An alternative view, more recently advocated by investigators who have examined partial 18S ribosomal RNA data, proposes that annelids, molluscs, and certain other minor phyla with trochophore larva stages share a more recent common ancestor with one another than any do with arthropods. The two hypotheses are mutually exclusive in explaining spiralian relationships. Cladistic analysis of morphological data does not reveal phylogentic relationships among major spiralian taxa but does suggest monophyly for both the annelids and molluscs. Distance and maximum-likelihood analyses of 18S rRNA gene sequences from major spiralian taxa suggest a sister relationship between annelids and molluscs and provide a clear resolution within the major groups of the spiralians. The parsimonious tree based on molecular data, however, indicates a sister relationship of the Annelida and Bivalvia, and an earlier divergence of the Gastropoda than the Annelida–Bivalvia clade. To test further hypotheses on the phylogenetic relationships among annelids, molluscs, and arthropods, and the ingroup relationships within the major spiralian taxa, we combine the molecular and morphological data sets and subject the combined data matrix to parsimony analysis. The resulting tree suggests that the molluscs and annelids form a monophyletic lineage and unites the molluscan taxa to a monophyletic group. Therefore, the result supports the Eutrochozoa hypothesis and the monophyly of molluscs, and indicates early acquisition of segmented body plans in arthropods. Received: 25 September 1995 / Accepted: 15 March 1996     

不同垦复方式油茶林土壤节肢动物多样性特征

【目的】油茶新造林特别是纯林不断骤增导致油茶林害虫发生日趋严重,食用茶油安全倍受威胁。为探索有效控制油茶林害虫发生的途径,保护分解者跳虫等土壤节肢动物多样性。【方法】基于Tullgren干漏斗法和巴氏罐法,对半垦复油茶幼林、全垦复油茶幼林、全垦复油茶成林和未垦复油茶成林4种不同油茶林的20个土样和15个凋落物样中的土壤跳虫和土壤其他节肢动物多样性特征进行了研究。【结果】共分离出跳虫315头,隶属6科8属;其他土壤节肢动物11科17种295头。多样性分析结果表明,油茶成林地比幼林地土壤动物的多样性指数高,多样化的地被物有利于提高土壤动物的多样性,垦复和施肥等人为干扰显著地降低土壤跳虫的多样性,却有利于增加罐诱土壤动物的多样性,水源是土壤动物生存的关键因子,对林地跳虫和罐诱动物的多样性提高有显著促进作用。【结论】探明了不同垦复类型油茶林土壤跳虫和土壤其他罐诱节肢动物多样性特征,对合理开展油茶林垦复,发挥油茶林土壤跳虫和土壤节肢动物分解功能提供了科学依据。     

Role of arthropod communities in bioenergy crop litter decomposition†

The extensive land use conversion expected to occur to meet demands for bioenergy feedstock production will likely have widespread impacts on agroecosystem biodiversity and ecosystem services, including carbon sequestration. Although arthropod detritivores are known to contribute to litter decomposition and thus energy flow and nutrient cycling in many plant communities, their importance in bioenergy feedstock communities has not yet been assessed. We undertook an experimental study quantifying rates of litter mass loss and nutrient cycling in the presence and absence of these organisms in three bioenergy feedstock crops—miscanthus (Miscanthus x giganteus), switchgrass (Panicum virgatum), and a planted prairie community. Overall arthropod abundance and litter decomposition rates were similar in all three communities. Despite effective reduction of arthropods in experimental plots via insecticide application, litter decomposition rates, inorganic nitrogen leaching, and carbon–nitrogen ratios did not differ significantly between control (with arthropods) and treatment (without arthropods) plots in any of the three community types. Our findings suggest that changes in arthropod faunal composition associated with widespread adoption of bioenergy feedstock crops may not be associated with profoundly altered arthropod‐mediated litter decomposition and nutrient release.     

Traits reveal ecological strategies driving carrion insect community assembly

1. The succession of carrion-associated (necrophilous) insects on decomposing carrion is well documented. To exploit the changing nutritious and dynamic resources available throughout the carrion decomposition process, different species colonise and consume carrion in a predictable temporal sequence. The traits of these necrophilous insects should reflect their ecological strategies. Morphological traits of these insects, such as body size and wing size, however, have not previously been examined during active and advanced decomposition. 2. We used fourth-corner multivariate generalised linear models to identify insect community morphological trait patterns and to quantify their change through time on decomposing rabbit carcasses in grassland and woodland environments. 3. We found that larger-bodied species of flies and carrion-specialist beetles were associated with the early stages of decomposition. The morphological traits of ants, in contrast, showed no changes at carcasses through time and instead showed body size differences between grassland and woodland environments. 4. Our findings indicate that specialist flies and beetles that arrive early in the decomposition process possess traits that enable rapid discovery of carrion at a large scale. Generalist beetles and ants do not share this same trait and are instead adapted to locate and consume a wider variety of resources in their preferred habitat type at their local scale. 5. Our results provide insights into the morphological adaptations linked to the ecological strategies of distinct components of carrion insect communities. Further, our results offer insights into the community assembly dynamics that structure the communities of necrophilous insect species.