The use of bacteriological analysis of the blood in the diagnosis of death by drowning

Investigation of blood samples from the femoral vein of subjects drowned in water and died of other causes (control) was carried out. Penetration of microflora contained in water into the bloodflow of the drowned people was shown. Possibility of using bacteriologic analysis of the postmortal blood to diagnose death from drowning was stated. Ps. putida and Ps. fluorescens. that were not present in the blood of persons who had died of causes other than drowning were recommended as test bacteria. Selective media and methods to isolate the bacteria from the cadaveric blood were chosen.     

PCR-DHPLC法检测硅藻SSU基因在溺死鉴定中的应用

目的采用PCR-DHPLC法检测硅藻SSU基因,评估其在溺死鉴定中的应用价值。方法 60只实验兔随机分为生前溺死（水中溺毙）、死后入水（空气栓塞致死后入水）、对照组（空气栓塞致死后不做处理）;溺死人体脏器组织;取各组织检材提取硅藻DNA,PCR扩增硅藻特异的核糖体小亚基（SSU）片段,用琼脂糖凝胶电泳检测、DHPLC检测分析。结果 6份硝酸消化法检测阴性的溺死人体器官组织检材经PCR及琼脂糖凝胶电泳检出5例阳性。生前溺死组肺、肝、肾硅藻检出率分别为100%、90%、85%,死后入水组仅肺检出硅藻（15%）,对照组各组织均为阴性;生前溺死组检出率明显高于死后入水组（P〈0.05）。10份溺死人体器官组织检材采用DHPLC法检出硅藻种类明显多于微波消解-扫描电镜法（P〈0.05）。脏器检出硅藻种类与溺死点水样一致。结论采用PCR-DHPLC法检测硅藻SSU基因,有助于溺死鉴定和溺死地点的推断,具有法医学应用价值。     

Comparison between lung weight and blood strontium in bodies found in seawater

This paper examines the use of lung weight increase as an indicator of seawater drowning compared to the amount of Sr absorbed by the blood. The study population was limited to male victims older than 20 years. Significant differences between cases of drowning and "non-drowning" were detected in terms of the lung-heart weight ratio (L/H) (p<0.001) or lung-body weight ratio (L/B) (p=0.005). However, using lung weight (L), L/H or L/B to distinguish between seawater drownings and saltwater non-drownings some overlap was produced. The factor rendering least overlap was L/B, which also appeared to be non-dependent on the victim's age. Our findings suggest that a value of L/B higher than 19.5 g/kg could be a useful indicator of death by drowning, but that when a lower value is found, additional drowning diagnoses would be needed to establish the manner of death.     

Selective culturing and genus-specific PCR detection for identification of Aeromonas in tissue samples to assist the medico-legal diagnosis of death by drowning

The detection of autochthonous aquatic bacteria in tissue samples from drowning cases is increasingly considered as an alternative approach to assist the medico-legal diagnosis of death by drowning. Bacteria belonging to the genus Aeromonas may be suitable candidates for this application as they are ubiquitous in natural aquatic environments but are generally not part of the human microbiota. The research aims of this study were (i) to develop a sensitive, specific and rapid screening and confirmation method for Aeromonas species in tissue samples and (ii) to evaluate aseptic sternal puncture as a post-mortem sample technique and bone marrow as an alternative matrix to provide evidence of death by drowning. The presence of Aeromonas in tissue samples was verified by cultivation using the selective media Ampicillin Dextrin Agar (ADA) and Ryan's Aeromonas Medium. The use of ADA medium was found most optimal for the sensitive, inexpensive and quick detection of aeromonads in human tissue samples. Positive culture plates were confirmed by harvesting all colonies for DNA extraction and subsequent PCR amplification using Aeromonas genus-specific primers. Aeromonads were detected in lung swab, blood and bone marrow of drowned bodies (n=3), but were negative in these three matrices for all negative controls (n=90) tested. Bone marrow proved to be a suitable alternative matrix and can be sampled post-mortem by an aseptic sternal puncture. In conclusion, this study confirms previous indications that aeromonads in cultures from blood of water bodies can be considered a potential marker for drowning. Given the fact that the number of immersed bodies (drowned and non-drowned) included in this study is statistically not significant, however, more tissue samples need to be investigated to confirm the validity of these methods to aid the diagnosis of death by wet drowning.     

溺死诊断中浮游生物检验方法的研究现状及进展

法医学实践中,判断是否为“生前溺死”是法医学尸检及鉴定工作中的一个重点和难点,目前尚无能明确诊断溺死的生物学和法庭化学指标。在溺死案件的诊断中,浮游生物检验是常见的方法。尽管目前还存在一些争议,但国内法医工作者认为,器官组织中浮游生物的存在仍然是判断生前溺死的重要证据之一。本文就国内外有关浮游生物的检验方法、检验标准及...     

Blood strontium concentration related to the length of the agonal period in seawater drowning cases

The levels of ventricular blood strontium (Sr) from 70 seawater drowning victims were compared with their diagnosis of drowning based mainly on certain criteria selected from their autopsy report. From this comparison, intervals of either the difference of Sr concentration between the left and the right ventricle blood (LVSr-RVSr) or the Sr concentration in the left ventricle blood (LVSr), appear to be related to different time-lapses of the agonal period of drowning. In the aim to diagnose drownings, intervals of both LVSr-RVSr and LVSr were proposed to characterize three different agonal periods in seawater drowning cases: instantaneous death (ID), fast vital-submersion drowning (FVSD) and common vital-submersion drowning (CVSD).     

Postmortem serum uric acid and creatinine levels in relation to the causes of death

Serum uric acid (UA) and creatinine (Cr) mainly derive from skeletal muscle tissues. Although, remarkable postmortem stability of the serum levels has been reported, there appears to be very poor knowledge of the diagnostic value in investigation of death, except for uremia. The aim of the present study was to evaluate postmortem serum UA and Cr levels using 395 forensic autopsy cases, in comparison with blood urea nitrogen (BUN), for investigation of the pathophysiology of death with special regard to the causes of death involving possible skeletal muscle damage, e.g. due to hypoxia, heat or agonal convulsions. Cr and BUN showed relatively good topographic stability in the cadaveric blood, whereas, UA was often much higher in the right heart blood than in the left heart and peripheral blood, independent of postmortem intervals. Moderate to marked elevation of Cr and BUN accompanied with hyperuricemia was observed in delayed death. In the acute death cases (survival time <30 min), UA, especially in the right heart blood, showed a considerable elevation in mechanical asphyxiation and drowning. The Cr level in fire victims with a lower carboxyhemoglobin (COHb) level (<60%) was significantly higher than in those with the possible fatal level (>60%). A similar elevation of Cr was observed in fatalities from heat stroke and methamphetamine (MA) poisoning. The observations suggested that hyperuricemia in acute death may be indicative of advanced hypoxia and that elevated Cr level may reflect the skeletal muscle damage, especially due to thermal influence.     

溺死诊断中浮游生物检验方法的研究现状及进展

法医学实践中,判断是否为生前溺死是法医学尸检及鉴定工作中的一个重点和难点,目前尚无能明确诊断溺死的生物学和法庭化学指标。在溺死案件的诊断中,浮游生物检验是常见的方法。尽管目前还存在一些争议,但国内法医工作者认为,器官组织中浮游生物的存在仍然是判断生前溺死的重要证据之一。本文就国内外有关浮游生物的检验方法、检验标准及检验结果的判定等研究现状作一综述,旨在使人们更加清晰地认识浮游生物检验在溺死诊断中的作用。     

浮游生物16S rDNA检测在大鼠溺死鉴定中的价值

目的建立浮游生物16S rDNA PCR检脸法应用于溺死的鉴定。方法将15只SD大鼠随机分为溺死组、死后抛尸组和正常对照组,死后分别取脑、肝、肾和肺等脏器,提取DNA.用特异性引物扩增浮游生物16S rDNA第三和第四可变区特异片段。结果溺死组5例,肺全部检出(100%)浮游生物16S rDNA;肝、肾分别检出4例(80%);脑检出3例(75%)。死后抛尸组仅1例在肺中检出浮游生物(20%)。正常对照组均未检出。苗论浮游生物16S rDNA PCR检验法可用于溺死的鉴定。     

Limbs found in water: investigation using anthropological analysis and the diatom test

We report the investigation, using a multi-disciplinary approach, of five cases of dismembered limbs which were recovered from Lake Ontario, Lake Erie and the Niagara River, and examined at the Office of the Chief Coroner for Ontario. In all cases, postmortem examination revealed that the limbs had been disarticulated in the postmortem period, by non-human taphonomic processes. In addition to routine gross examination, the femur and/or tibia were assessed using anthropological methods to give estimates of the sex, age, race and stature of the individual. The anthropologic data facilitated the identification of one of the cases. In all cases, nitric acid extracts of the femoral bone marrow were prepared and examined for the presence of diatoms. In all instances, diatom frustules were recovered from marrow extracts, indicating that drowning was the cause of death or at least a significant contributing factor in the cause of death. The use of the diatom test was helpful in excluding the possibility that the limbs were dismembered from individuals who had died by means other than drowning, and had been subsequently 'dumped' into water. The application of anthropological methods and the diatom test for drowning may significantly enhance the medico-legal investigation of body parts recovered from water, and we present an overview of useful techniques here. Anthropological data may facilitate identification, and the diatom test may establish a cause of death.     

溺死的法医学检验现状及进展

本文概括阐述了近年来,国内外学者将硅藻的检验、内脏异物颗粒和微量元素成分的检验、血液化学的检验、电子显微镜的观察等方法应用于溺死的诊断,试图为溺死的法医学鉴定提供准确的诊断指标。     

Study of the diagnostic value of iron in fresh water drowning

The aim of our study was to test the diagnostic value of iron (Fe) in fresh water drowning by investigating the postmortem levels of hemodilution in drowning cases compared to control cases. Twenty-six typical fresh water drowning cases were selected from 128 immersion cases autopsied in our Department of Forensic Pathology between 1998 and 2004. The exclusion criteria were a long postmortem interval and other causes of death than drowning. For all selected cases, the diagnosis of drowning was based on the presence of autopsy findings and positive diatom test. A control population of 12 cases was also selected. For each drowning and control case, iron blood levels were measured in the left ventricle (LV) and right ventricle (RV) of the heart. The mean difference of iron concentration (RVFe-LVFe) between the drowning group and the control group was statistically compared. Furthermore, iron measurements were performed in 19 drowning cases showing advanced putrefaction. The mean difference of iron concentration was significantly higher in the drowning cases compared with controls (P<0.001). All drowning cases showed hemodilution. No overlap was found in the RVFe-LVFe levels between the two groups. Resuscitation attempts seemed to have no effect on the results. In cases of drowning showing advanced putrefaction, the iron test was not reliable because biochemical iron measurement was often prevented by no sufficient blood in the heart or postmortem clots. In conclusion, according to our results, iron seems to be a good biochemical marker in fresh water drowning with a short postmortem interval.     

A promising microbiological test for the diagnosis of drowning

A number of biological and chemical tests have been developed over the years to determine whether a person was drowned. This study focuses on the potential of a microbiological test for detecting common bacterial markers of water faecal pollution such as faecal coliforms (FC) and faecal streptococci (FS) as possible indicators of drowning. A promising previous study was carried out on central and peripheral blood samples of 42 drowned victims (20 cases in saltwater and 22 cases in freshwater) and 30 not-drowned bodies. To improve the accuracy of our previous results and also in order to investigate a possible cause of a false positive due to pulmonary passive diffusion and subsequently endogenous or exogenous bacterial invasion of the blood in the post-mortem interval (PMI), the FC and FS test was applied to bodies submerged in water but died from causes other than drowning. In the present study, blood samples collected from the left ventricle (LV), right ventricle (RV), femoral artery (FA) and, femoral vein (FV) of 10 drowned victims (5 cases in freshwater and 5 cases in seawater) and 3 not-drowned individuals with bodies submerged in water for a while after death have been analysed. Preliminary results are in agreement with other reports dealing with diatoms and marine bacteria that suggest to exclude the hypothesis of a passive penetration of sufficient quantities of drowning medium into circulation after death or during the agonal period. Based on our results there is also no evidence of a relevant dissemination of endogenous micro-flora from the gastrointestinal tract affecting the FS and FC test. There are still several other factors that could influence the applicability of post-mortem FS and FC cultures for the diagnosis of drowning and they need further investigations. The present article provides only a glimpse of the potential of the FS and FC test as bacteriological method for the diagnosis of drowning.     

水中尸体PMCT最新研究进展

溺水是公安法医日常检案中最常见的死因之一,水中发现的尸体是否为生前入水,是公安法医日常鉴定的要点也是难点,而一旦溺亡的案件中掺杂了其他复杂的案情,其鉴定意见失真也极易引起不良的社会影响。法医鉴定过程中,溺死一般依据尸体征象、组织病理学检验和硅藻检验等综合考虑。PMCT为法医诊断溺死提供了新的思路并被许多国家和地区常规使用于诊断溺死。本文综述了国内外近年来使用PMCT对溺死尸体呼吸系统、消化系统和循环系统等方面的研究成果,讨论了PMCT诊断溺死的应用价值与潜力,并对国内PMCT发展的必要性和前景进行了展望。     

Diatoms in forensic expertise of drowning--a Macedonian experience

Results obtained by examination of 22 human cases suspected for drowning, one human case of death other than drowning and several tests on laboratory rats were used as a basis for evaluation of diatom method as supportive in forensic expertise of drowning. The recovery of diatoms from various examined organs, their qualitative and quantitative composition, if properly treated without the possibility of contamination, can be a reliable proof of the time and place of drowning. The priority of organ examination (external microflora determination, lungs, brain, heart (and/or blood), stomach, liver and kidney, and finally bone marrow) is discussed and established as well as the basic future research on cases suspected of drowning, but also on non-drowned victims and laboratory animals.     

Diagnosing death by drowning in fresh water using blood strontium as an indicator

The interpretation of the level of strontium (Sr) in blood has been demonstrated to be useful in the diagnosis of death by drowning occurred in sea water, but its use in fresh water drownings is not so evident because of the low Sr concentration present in most of fresh water media. In this paper, we show a survey of the results obtained in the casework analysis of Sr in ventricular blood used in the diagnosis of drowning in 144 bodies found immersed in fresh water over a period of 10 years. Thirty-two percent of the immersion cases examined could be diagnosed as drownings with a reasonable degree of confidence. It is thought that this percentage of positive diagnoses could be largely improved in the case of blood samples taken a few hours after death.     

非溺死尸体肺脏硅藻最大值在溺死鉴定中的应用初探

探讨非溺死尸体肺脏硅藻最大值（即空气吸入与检验污染之和的最大值）在溺死鉴定中的应用价值。用硝化-光镜法检测贵阳地区40例非溺死尸体肺脏硅藻含量的最大值，同时作了8例溺死尸体肺脏硅藻检验。非溺死组肺脏硅藻最大值2个／5g；溺死组有7例为56～463个／5g，1例是2个／5g。表明（1）水中尸体肺脏硅藻检验值大于最大值（2个／5g），可考虑为溺死；显著大于最大值（本文溺死组7例56～463个／5g）则可鉴定为溺死；（2）等于或小于最大值，提示干性溺死或抛尸入水，可结合尸检发现及案情鉴别。     

Interpretation of the term "hydrocution"

Based on a literature study, several interpretations of the term "hydrocution" are presented and discussed. It is difficult to differentiate between hydrocution and atypical drowning. Therefore, the history, the pathogenetic mechanism and the autopsy findings must be considered to establish a clear diagnosis when assessing a sudden death in the water. Practicable diagnosis criteria are recommended.     

检测浮游生物叶绿素相关基因诊断溺死的实验研究

目的评估浮游生物叶绿素相关基因检测用于溺死诊断的价值。方法将18只大白兔随机分为溺死组（n=10）、死后抛尸组（n=6）和对照组（n=2）,各组分别取心血及肺、肝、肾、脑组织,分离浮游生物并提取其DNA,用PCR技术检测叶绿素相关基因EG（EG1和EG2）及SK（SK1和SK2）。同时用硝酸消化法检验肺和肝组织中的硅藻。结果溺死组心血及肺、肝、肾、脑组织中EG1分别检出9、10、9、7和8例阳性,EG2分别检出8、10、7、5和7例阳性;死后抛尸组仅在心血和肺组织中各检出1例EG1阳性;对照组各组织均未检出EG1和EG2。SK1、SK2除在溺死组心血、肺和肾有少数检出外（≤2例）,在其他组未检出扩增产物。硝酸消化法从溺死组肺、肝组织中分别有9例及3例检出硅藻,死后抛尸组仅在1例肺组织中检出。结论浮游生物叶绿素相关基因EG用于溺死诊断的阳性检出率要高于硝酸消化法,在溺死诊断中具有较高的应用价值。     

A study on the diagnosis of drowning by examination of lung chlorophyll(a) of planktons with a spectrofluorophotometer.

An investigation on the diagnosis of drowning by analysis of chlorophyll(a) (Chl.(a)) of planktons in the lung sample with a spectrofluorophotometer (SFPM) is reported. The diagnostic criteria or main points of death by drowning are suggested are according to our experimental studies. Meanwhile two cases of submersion in water were investigated and the causes of death were diagnosed correctly. It is concluded that examination of lung Chl.(a) of planktons could be used as a reliable diagnostic method in forensic practice.