Pig decomposition in lotic aquatic systems: the potential use of algal growth in establishing a postmortem submersion interval (PMSI)

While algal community composition has been examined as a qualitative indicator of postmortem submersion interval (PMSI), there have been no quantitative studies on using algal growth rates as PMSI estimators. The present study was undertaken to examine pig decomposition in streams and to develop a more quantitative approach to estimate a PMSI. Pigs and ceramic tiles were completely submerged and regularly sampled for periphyton growth. Five stages of decomposition were identified for the submerged pig carcasses according to physical characteristics. Algal growth rates, measured quantitatively as a function of chlorophyll-a concentration, were greater on pigs compared with tiles; however, microhabitat (pools versus riffles) did not significantly influence algal growth. Additionally, there was a strong correlation between algal growth rate and time on pigs and tile substrates. This strong correlation was observed after significant rain events. Our study documents for the first time a quantitative technique to determine the length of time a corpse has been submerged in water. We suggest that algal growth rates may be a useful quantitative indicator in criminal investigations involving corpses that are completely submerged in stream or riverine habitats.     

The Potential of High-throughput Metagenomic Sequencing of Aquatic Bacterial Communities to Estimate the Postmortem Submersion Interval

Human remains can be discovered in freshwater or marine ecosystems, circumstances where insects and other invertebrates have infrequently been used for understanding the time of postmortem submersion. In this study, the identification and succession of epinecrotic bacterial communities on vertebrate remains were described during decomposition in a temperate headwater stream during two seasons (summer and winter). Bacterial communities were characterized with 454 pyrosequencing and analyzed at phyletic and generic taxonomic resolutions. There was a significant increase in genera richness over decomposition during both seasons. Additionally, multivariate statistical modeling revealed significant differences in bacterial communities between seasons at both taxonomic resolutions and siginificant genera differences among sampling days within each season, suggesting a succession of these communities. These data are the first to describe aquatic bacterial succession using high-throughput metagenomic sequencing on vertebrate remains submerged in a freshwater habitat, and provide initial evidence for their potential use in forensic investigations.     

Assessment of post-mortem submersion interval using total aquatic decomposition scores of drowned human cadavers

Investigating drowning-related deaths remains a significant problem for forensic personnel all over the world. The previously published decomposition scoring method like the total aquatic decomposition (TAD) score promises to estimate the correct post-mortem submersion interval (PMSI) in aquatic habitats through the assessment and calculation of the decomposition rate and accumulated degree days (ADD). The current study comprised of 53 drowned death cases belonging to various districts of Haryana from May 2016 to August 2017. The regression and Pearson's correlation indicated a significant correlation between the TAD scores and the actual ADD (calculated through water temperatures) (r² = 0.917) and between the actual and the estimated ADDs (calculated through TAD scores used by Heaton et al. [21]) (r = 0.9585). The results indicated that the estimated ADD tends to over predict the PMSI compared to the actual ADD. It is further confirmed by paired t-test, which showed the mean of actual ADD (mean = 349) to be significantly lower than the mean of estimated ADD (mean = 663). Moreover, these methods will help forensic investigators and researchers formulate region-specific regression equations for PMSI estimation.     

An Aquatic Decomposition Scoring Method to Potentially Predict the Postmortem Submersion Interval of Bodies Recovered from the North Sea

This study aimed to develop an aquatic decomposition scoring (ADS) method and investigated the predictive value of this method in estimating the postmortem submersion interval (PMSI) of bodies recovered from the North Sea. This method, consisting of an ADS item list and a pictorial reference atlas, showed a high interobserver agreement (Krippendorff's alpha ≥ 0.93) and hence proved to be valid. This scoring method was applied to data, collected from closed cases—cases in which the postmortal submersion interval (PMSI) was known—concerning bodies recovered from the North Sea from 1990 to 2013. Thirty‐eight cases met the inclusion criteria and were scored by quantifying the observed total aquatic decomposition score (TADS). Statistical analysis demonstrated that TADS accurately predicts the PMSI (p < 0.001), confirming that the decomposition process in the North Sea is strongly correlated to time.     

Comparison of Protocols for Measuring and Calculating Postmortem Submersion Intervals for Human Analogs in Fresh Water

Protocols for determining postmortem submersion interval (PMSI) have long been problematic for forensic investigators due to the wide variety of factors affecting the rate of decomposition of submerged carrion. Likewise, it has been equally problematic for researchers to develop standardized experimental protocols to monitor underwater decomposition without artificially affecting the decomposition rate. This study compares two experimental protocols: (i) underwater in situ evaluation with photographic documentation utilizing the Heaton et al. total aquatic decomposition (TAD) score and (ii) weighing the carrion before and after submersion. Complete forensic necropsies were performed as a control. Perinatal piglets were used as human analogs. The results of this study indicate that in order to objectively measure decomposition over time, the human analog should be examined at depth using the TAD scoring system rather than utilizing a carrion weight evaluation. The acquired TAD score can be used to calculate an approximate PMSI.     

Use of aquatic insects in determining submersion interval

Although its potential is great, the use of aquatic insects in determining submersion intervals at death-scene investigations has not been exploited in the past. Aquatic environments have no known true specific indicator species, as do terrestrial habitats. However, aquatic environmental studies show that organisms may colonize a substrate dependent on factors such as size, position, exposure to current, water temperature, current speed, water depth, the presence of algal communities, or detritus. Certain aquatic insects such as the chironomid midges (Diptera, Chironomidae), and the caddisflies (Trichoptera), are capable of colonizing immersed bodies; and with the known biology of a specific species of insect for a certain geographic area, time intervals of submersion can be established.     

Marine bacterial succession as a potential indicator of postmortem submersion interval

The process of decomposition of bodies in the marine environment is poorly understood and almost nothing is currently known about the microorganisms involved. This study aimed to investigate the microbes involved in decomposition in the sea and to evaluate the potential use of marine bacterial succession for postmortem submersion interval (PMSI) estimation, for which there is currently no reliable method. Partial pig remains were completely submerged during autumn and winter and were regularly sampled to document marine bacterial colonisation and the changes in community composition over time. Five stages of decomposition were recognised, some of which exhibited characters specific for partial carrion. Marine bacteria rapidly colonised the submerged remains in a successional manner. Seasonal differences were observed for the rate of decomposition and also for several groups of colonising bacteria. Marine bacteria specific for particular PMSIs were identified. This study provides an insight into the involvement of saprophytic marine bacteria in the decomposition of mammalian remains in the sea and is the first to explore the use of marine bacterial colonisation and succession as a novel tool for PMSI estimation. We propose that with further study, marine bacterial succession will prove useful for determination of the length of time a body may have been immersed in a marine environment.     

The "coffin fly"Conicera tibialis (Diptera: Phoridae) breeding on buried human remains after a postmortem interval of 18 years

The "coffin fly,"Conicera tibialis Schmitz (Order: Diptera, Family: Phoridae), is well known for its frequent occurrence on buried corpses, in some cases after postmortem intervals of even 3-5 years. The present report describes the presence of a large amount of individuals of C. tibialis inside the coffin of a buried human corpse exhumed 18 years after death in central Spain. Adults, some of them newly emerged, and empty puparia were found in connection with the remains. Such postmortem interval is significantly longer than previously known for this species and raises the question on the current state of knowledge about the use of insects for estimating the postmortem interval in old, buried remains.     

The potential to determine a postmortem submersion interval based on algal/diatom diversity on decomposing mammalian carcasses in brackish ponds in Delaware

Recently botanical evidence has been studied to determine if it is useful in forensic investigations. This study was performed to examine stillborn piglet decomposition in a brackish water environment and to semi-quantitatively document stages of decomposition, degree day accumulation per stage as well as the algal/diatom diversity useful in determining a postmortem submersion interval (PMSI). Piglets and ceramic tiles were submerged in brackish ponds and sampled on a regular basis to document algal diversity and succession between substrates and stages of decomposition. Significantly greater weight was lost from piglet carcasses during the early floating and advanced floating decay stages. Seasonal effects were observed in degree-day accumulations. Diatom diversity was significantly greater on piglet carcasses compared to tile substrates. Algal diversity decreased over time on the piglet carcasses as well as the stage of decomposition. A significant relationship and strong correlation between algal diversity found on the piglet substrate with time was observed. Our results indicate that more research is needed to examine the potential to use diatoms in not only determining manner of death but also the duration of time (PMSI) a victim may have been immersed in an aquatic environment.     

Taphonomy of child-sized remains: a study of scattering and scavenging in Virginia, USA

Child-sized pig carcasses (Sus scrofa) were placed in surface deposit and buried scenarios in a wooded area of Virginia from May 1998 through December 2000, to examine the taphonomic effects of decompositional changes, predator scavenging, and the extent of remains scattering. Changes were observed through on-site examination, charting of remains, and recorded video imaging. Analysis of data revealed that utilization of corpses as food sources by vertebrates was dependent upon invertebrate colonization. Vertebrates avoided feeding on the corpses while invertebrate colonization was active, and would feed before invertebrates successfully colonized a corpse, or would wait until the invertebrate populations migrated away from the corpse. Among vertebrates, there was no apparent succession order for the animals utilizing the remains as a food source. Different vertebrates would feed at different times based upon diurnal or nocturnal predilection. Analysis noted an accidental cooperative relationship between the invertebrates and vertebrates scavenging on the corpses. Certain vertebrates gained access to the internal tissues by utilizing openings in the corpses caused by invertebrate and other vertebrate scavenging. Alternately, carrion-frequenting insects were afforded access to previously inaccessible colonization sites as a result of scavenging vertebrate activities.     

新乡地区夏季常见嗜尸性昆虫群落演替规律初步研究

目的研究新乡地区夏季嗜尸性昆虫的种类及其群落演替规律。方法观察法研究新乡医学院动物房后偏僻草地上6只大白鼠和5只家兔尸体腐败过程中嗜尸性蝇类的主要物种及其群落演替规律。结果该地区主要出现嗜尸性蝇类为双翅目3科7种,包括家蝇、大头金蝇、丝光绿蝇、棕尾别麻蝇、黑尾黑麻蝇、厩腐蝇、元厕蝇,且呈现出演替规律性。结论研究结果可作为新乡地区对尸体死亡时间的推断依据。     

Practice of forensic entomology--usability of insect fragments in the estimation of the time of death

Forensic entomology analyzes and evaluates the presence of necrophagous or necrophilic insects on human corpses to estimate the post-mortem interval. The three cases presented are good examples illustrating that not only the various stages of development of the necrophagous insects from egg via larva to pupa are useful for forensic assessment. Depending on the situation in the individual case, even inconspicuous remains of past insect activity such as empty pupal cases or larval skins may supply important entomological information. For this purpose it is indispensable to preserve such evidence, which may appear only as fragments or pieces at first sight, for proper entomological assessment.     

Biolog-Eco法检测尸体微生物群落的代谢功能变化

目的检测死后不同时间尸体上微生物群落功能多样性的变化并评估其在死亡时间推断中的应用价值。方法采用Biolog-Eco微平板对野外实验中来自于死后0~240 h猪和人尸体的肛门拭子进行微生物群体的培养,监测其引起的光密度值变化,结合法医病理学及蝇类演替,观察自然腐败尸体微生物群落的代谢特性及其变化。结果微生物代谢功能多样性与蛆虫数量变化呈负相关关系。冷冻在0 h对每孔颜色平均变化率影响最大,48 h后基本消失,192 h后尸体微生物群落多样性相对不稳定。主成分分析将31种碳源综合为5个主成分(累积贡献率90%)。碳源tsquare分析显示,N-乙酰-D-葡萄糖氨和L-丝氨酸是推断人和猪样品PMI(0~240 h)的优势碳源。结论 Biolog-Eco法能表现死亡后0~240 h尸体上微生物群落对部分碳源利用的代谢差异,有望为死亡时间推断提供新依据。     

鉴定溺死的实验研究——荧光分光光度计检测肺组织浮游生物叶绿素(a)

本实验应用荧光分光光度计检测肺组织浮游生物叶绿素(a),以期通过准确的定性、定量研究来诊断溺死。实验性研究表明本法准确可靠,选择性强、灵敏度高,不受腐败及污染等因素影响,亦不存在含叶绿素(a)浮游生物生前进入并存在于人体器官的问题。同法对2例水中尸体进行了检测,结果与实验性研究相符、提示本法可应用于实际检案。     

Soft Tissue Decomposition of Submerged,Dismembered Pig Limbs Enclosed in Plastic Bags*

Abstract: This study examines underwater soft tissue decomposition of dismembered pig limbs deposited in polyethylene plastic bags. The research evaluates the level of influence that disposal method has on underwater decomposition processes and details observations specific to this scenario. To our knowledge, no other study has yet investigated decomposing, dismembered, and enclosed remains in water environments. The total sample size consisted of 120 dismembered pig limbs, divided into a subsample of 30 pig limbs per recovery period (34 and 71 days) for each treatment. The two treatments simulated non‐enclosed and plastic enclosed disposal methods in a water context. The remains were completely submerged in Lake Ontario for 34 and 71 days. In both recovery periods, the non‐enclosed samples lost soft tissue to a significantly greater extent than their plastic enclosed counterparts. Disposal of remains in plastic bags therefore results in preservation, most likely caused by bacterial inhibition and reduced oxygen levels.     

Temperature‐dependent Development of Parasarcophaga similis (Meade 1876) and its Significance in Estimating Postmortem Interval

Flesh flies are commonly found insects on decaying corpses that appears slightly later than blowflies, and their development patterns are significant indicators for minimum postmortem interval (PMI_min) estimation. In this study, the flesh fly Parasarcophaga similis (Meade 1876) was reared at nine constant temperatures ranging from 15°C to 35°C to examine indicators for estimating their age. We generated three development models, including isomorphen diagram, isomegalen diagram, and thermal summation model. Larval body length at different rearing temperatures was fit into an L = a + bT + cT² + dT³ equation with which the relationship between the larval body length (L) and the time after larviposition (T) was confirmed. The pupal stage was categorized into 13 substages according to intrapuparial morphological changes, and a detailed table was generated of the pupal developmental stages at five rearing temperatures, 15°C, 20°C, 25°C, 30°C, and 35°C. This study provides fundamental data in supporting P. similis as an indicator for PMI_min estimation.     

Initial study of arthropods succession and pig carrion decomposition in two freshwater ecosystems in the Colombian Andes

Entomological succession and trophic roles of arthropods associated with different stages of carcass decomposition were studied to estimate the post-mortem submersion interval in two freshwater ecosystems in the Colombian Andes, at an altitude of 2614 m. Pig carcasses were employed as models placed 68 m apart, one in a stream (lotic) and another in an artificial lake (lentic). Decomposition time to skeletal remains was 74 days in the lake and 80 days in the stream. Six phases of decomposition were established: submerged fresh, early floating, floating decay, bloated deterioration, floating remains and sunken remains. A total of 18,832 organisms associated with the carcasses were collected: 11,487 in the lake (four orders, 19 families and 33 species) and 7345 in the stream (eight orders, 15 families and 25 species). Organisms were classified in the following ecological categories: shredders, collectors, predators, necrophagous, sarcosaprophagous and opportunists. Physical and chemical properties of the habitats, such as water temperature, CO(2) and conductivity, varied according to rainfall. In the lake, shredders (Coleoptera: Tropisternus sp. and Berosus sp.) and collectors (Diptera: Chironomus sp.) were found to be associated with submerged phases. Predators (Odonata) were only present during the first phases. Coleoptera (Dytiscidae) were found during floating decay and bloated deterioration stages. In the stream, shredders (Hyalella sp.) and collectors (Simulium sp.) were found during all stages, whereas the predator Oxelytrum discicolle was found exclusively during the floating stages, during which body temperature increased in a fashion similar to active decay in terrestrial environments.