Serum biochemical markers in drowning: diagnostic efficacy of Strontium and other trace elements

The aim of our study was to analyse several serum biochemical markers in order to evaluate the discriminant capacity of trace elements individually and jointly in drowning (seawater drowning and freshwater drowning) and their correlation with age, sex, postmortem interval, time in water and concentrations of the trace elements in the drowning medium. Sixty-seven cases of drowning (53 seawater drownings (SWD); 14 freshwater drownings (FWD) and 73 control cases (other asphyxias, n=44, and other causes of death, n=29) were selected according to the scene, cause and circumstances of death, together with autopsy findings. Serum strontium (Sr), magnesium (Mg), sodium (Na), chloride (Cl), calcium (Ca), iron (Fe), urea, creatinine (Cr) and cardiac Troponine T (cTn-T) were measured in the left ventricle (Lv), right ventricle (Rv) and peripheral blood. Lv-Rv differences for each marker and Sr, Mg, Na, Cl, Ca and Fe concentrations in the drowning medium were determinated. Mean concentrations of Sr, Cl and Mg in both ventricles and peripheral serum and Lv-Rv differences and Ca Lv and Na Rv were significantly higher in cases of drowning than for other causes of death. In SWD, Sr, Mg, Ca, Na and Cl were significantly higher in Lv than in Rv as a result of aspirating water. In contrast, haemodilution is evident from the significantly higher levels of Fe and urea in Rv than in Lv in cases of SWD, and from the higher Mg and Cr levels in Rv in FWD. In the case of SWD, serum levels of Sr are confirmed as the best parameter for diagnosis, although other trace elements may also be useful, such as the serum concentrations of Mg and Cl. In the case of FWD, the joint determination of Sr and other biochemical markers, especially Fe, may increase correct diagnosis.     

Plasmatic levels of atrial natriuretic peptide (ANP) in drowning. A pilot study

Bodies found in water may cause problems for forensic pathologists who have to differentiate drowning from postmortem immersion or fresh from salt water drowning. The exact physiopathology of drowning is still controversial and complementary tests can not exactly establish the exact cause of death if macroscopic findings at autopsy are not conclusive. We have employed atrial natriuretic peptide (ANP) as a marker in an experimental series of fresh and salt water drowning, comparing their results with a non-drowned control series. There are differences between the plasma basal levels of the control series (79 pg/ml) and the levels in animals drowned in fresh water (358 pg/ml, P less than 0.001) and between control and rabbits drowned in salt water (190 pg/ml, P less than 0.001). According to these values, there are also differences between fresh and salt water drowned animals (P less than 0.001). We propose this peptide as a new marker in cases of drowning, with the ability to differentiate drowning from postmortem immersion and fresh from salt water drowning.     

测定肺锶含量鉴定溺死的研究

作者借助ICP 检测溺死和死后沉入水中,浸泡不同时间家兔及对照组肺腮含量。结果表明,溺死组肺锶含量显著高于死后入水组和对照组(P<0. 01) ;无论溺死或死后沉入水中浸泡不同时间,家兔肺锶含量之间无明显差异(P>0. 05) ,提示肺锶含量受水中浸泡时间或腐败因素的影响并不显著。2例实际案例的肺锶含量测定支持实验性研究结果,说明肺锶含量测定可用于鉴定溺死,具有实用价值。     

Aquatic fatalities--a systematic retrospective analysis

In a retrospective analysis of the autopsy material (n = 5,767) of the Institute of Legal Medicine of the Hanover Medical School covering the period of 1998-2007, all aquatic fatalities were evaluated, categorized and systematically compared under epidemiological and forensic criteria. The total of 156 cases of death by drowning (2.7 % of all autopsies) included 38 bathtub drownings and 28 deaths in the water for which no pathological anatomical cause of death could be reliably demonstrated. A control group (n = 221) was investigated for the presence of aqueous liquid in the sphenoid sinuses and compared with the findings of the drowning cases without signs of putrefaction. About 16 % of the control cases had fluid in the sphenoid sinuses compared with 57.6 % in the drowning group. Most of the drowning victims were men (60.9 %), whereas in the group of bathtub drownings the majority were women. More than half of the drowning cases (n = 89) could be classified as accidents. The individual groups showed a different incidence of findings associated with drowning.     

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).     

The usefulness of lung surfactant phospholipids (LSPs) in the diagnosis of drowning

The authors have studied the usefulness of some lung surfactant phospholipids (LSPs) isolated from lung tissues as markers of drowning. Two different groups of rabbits were sacrificed by drowning in fresh and salt water, and their phospholipid compositions were compared with those of a non-drowned control series. For the phospholipids studied in lung lavages (phosphatidyl choline, phosphatidyl ethanolamine, and phosphatidyl glycerol) the proportions differed between the control group and the drowned group, and between the fresh-water and salt-water drowned animals. According to these results, the lipids we have analyzed can be employed as markers in forensic autopsies, where it is necessary to differentiate between death by drowning and postmortem immersion and between fresh-water and salt-water drowning. In lung tissue, only phosphatidyl choline and phosphatidyl inositol showed significative differences. These results also confirm that LSPs are strongly affected in drowning.     

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.     

测定血锶含量鉴定溺死的法医学意义

为评价血锶含量测定对诊断鉴定溺死的实用价值,作者用家兔进行实验,分别测定溺死与死后入水家兔左心血中锶含量。结果证实:1.死后较短时间内血锶含量测定确可鉴定溺死;2、死后24小时内仅可帮助鉴定溺死。实验表明水中浸泡时间因素对血锶含量确有显著影响。两例实际检案的应用观察,较实验性研究为佳,值得进一步研究。     

Hemolytic staining of the intima of the aortic root in freshwater drowning: a retrospective study

A retrospective study was conducted of files at the Institute of Legal Medicine in Hamburg, Germany from 1998 to 2003 of all cases of drowning where there was no putrefaction, to document the rate of recording of hemolytic staining of the intima of the great vessels. All cases had full police investigations with reviews of previous histories, circumstantial evidence, and autopsy findings. A series of control subjects who had died of various natural and nonnatural, nondrowning deaths, matched for age, sex, and postmortem interval, was also reviewed. One hundred twenty cases of freshwater drowning were identified. Drowning occurred in a garden pool in 1 case, in a bath in 2, and in the river Elbe in 117. The age range was from 2 to 91 years (mean = 55 years; M:F = 1:1.8). Hemolytic intimal staining of the aortic root was documented in 6 cases (6 of 120; 5%). This consisted of reddened discoloration of the proximal portion of the aortic root, without any significant staining of the proximal pulmonary artery. No significant hemolytic staining of the intima of the great vessels was recorded in any of the 120 control cases. Although under-reporting of findings may occur in retrospective analyses, this study has shown that at least 5% of freshwater drowning cases showed differential staining of the pulmonary trunk and aorta, with hemolytic discoloration of the aortic intima. When present, hemolytic staining of the aortic root intima may be a useful and possibly under-recognized corroborative sign of freshwater drowning.     

Interpretation of postmortem change in cadavers in Spain

Estimating time since death is especially difficult in the examination of poorly preserved cadavers and depends on the experience of the examiner and comparison with previously documented cases showing similar characteristics. The present study reports on information obtained over the past ten years through the work of the Laboratorio de Antropología y Odontología Forense (LAF) of the Instituto Anatómico Forense de Madrid, Spain, in particular evaluating how the type of fracture influences postmortem change. From the original 225 forensic cases examined between 1992 and 2002 in the LAF, a sample of 29 cases were selected from various regions of the Spanish mainland. A data collection protocol was established to reflect factors which the existing specialized literature, documenting the relation existing in the sample analyzed between time since death and the extent of postmortem change, which in the environments examined are distributed into the following phases: Phase 1 (putrefaction): one week to one month on the surface and two months in water. Phase 2 (initial skeletonization): two months on the surface and five to six months in water. Phase 3 (advanced skeltonization): six months to 1.5 years on the surface and 2.5 years buried. Phase 4 (complete skeletonization): about one year on the surface and three years buried. This paper also provide useful information on the impact of carrion insect activity, location, climate, seasonality, and predator.     

PCR-DGGE法检测浮游生物16S rDNA在溺死鉴定中的应用

目的建立并评估PCR-DGGE法检测浮游生物对溺死鉴定的应用价值。方法大白兔30只随机分为3组:溺死组(n=12),死后抛尸组(n=12)和对照组(n=6);溺死组和死后抛尸组又分为2个亚组:东湖水域组(n=6)和墨水湖水域组(n=6)。死后提取心血和肺、肝、肾、脑等组织,匀浆后,采用Percoll密度梯度离心法分离浮游生物并提取其DNA,PCR扩增浮游生物特异的16S rDNA片段后分别用琼脂糖凝胶电泳及DGGE检测分析。2个溺死案例检材同法检验。结果溺死组各组织器官中浮游生物检测多呈阳性:肺(100%)、肝(83%)、肾(75%)、心血(83%)、脑(42%);死后抛尸组仅2例肺组织(16.7%)检出阳性;对照组全部阴性。溺尸肺组织DGGE分型图谱与相应溺死点水样分型图谱相似,而与非溺死点水样分型结果差异显著。2实际案例均呈阳性。结论本方法不仅有助于定性诊断溺死,而且通过比较产物的多样性可以推断溺死地点,在法医学溺死鉴定中具有较大实用价值。     

硅藻定量检验和种属鉴定的实验研究

本实验用家兔18只,分为溺死组、死后(抛尸)入水组及对照组,每组各6只。采用消化—光镜法作肺、肝、肾硅藻计数(定量)及种属鉴定检验,同时作水样检验及空白对照。结果:(一)进一步证实生前硅藻可经空气源性进入肺脏并存积于大循环内脏(肝、肾)的概念;(二)提示水中尸体肺脏如有大量硅藻检出,仅此即可作为诊断溺死的可靠依据;空气源性吸入及检验过程中的污染并不影响该检验结果对鉴定溺死的判断。种属分析溺死组肺与水样的主要硅藻种属相符,而肝、肾中硅藻计数及种属分析均无诊断意义。     

溺死法医学鉴定的研究新进展

综述了近年来在硅藻检验、水中浮游生物叶绿素（A）检测、血液化学和组织化学检验等方面的最新文献报道,并对各种溺死检验方法的优缺点进行了评价：在硅藻检验中,硝酸乙醇法、破机罐法及微波消解法,可缩短检验时间,提高办案效率;酶消化法及PCR法硅藻检出率高,适用于可疑水样中硅藻密度低等情况。早期器官组织中浮游生物叶绿素（A）、血液和组织中其他生化指标,可作为鉴定溺死的重要参考。微量元素锶检测可用于鉴定海水中溺死。另外,硅藻及其他浮游生物遗传多态性片断PCR,可望成为新的、灵敏的溺死检测方法。     

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。     

The forensic medical evaluation of the results of determining diatomaceous plankton in the diagnosis of drowning

Evaluation of demonstrative value of method based on diatom plankton determination as a diagnostical sign of death due to drowning was performed. Plankton was detected in the viscera in 37% of cases (as a whole 376 bodies of people who drowned in fresh water were examined). Plankton was detected in aspiration type of drowning and its mixed variant. In spastic and reflector types of drowning plankton wasn't detected. According to author's opinion this method is an objective diagnostical test to determine the cause of death of people when their bodies are found in water.     

Postmortem chest computed tomography for the diagnosis of drowning:a feasibility study

It may be difficult to distinguish the cause of death in drowning cases without specific findings. The aim of this study was to explore the forensic value of thoracic postmortem computed tomography (PMCT) using routine images and three-dimensional (3D) image reconstructions. The imaging data of PMCT examinations of six drowning cadavers, aged 21–54 years, were analyzed. Twelve victims of sudden death from coronary artery disease (CAD) were chosen as a control group. After 3D bilateral lung images were reconstructed using image processing software, an interactive medical image control system was used to measure and analyze parameters including lung volume, lung volume ratio, mean CT value of the whole lung, and lung CT value distribution curves. Lung volume and lung volume ratio were used to assess the shape changes of the lung. Lung CT value distribution curves showed the corresponding number of pixels of the different CT values in the lung image. Lung volume was not significantly larger in drowning cases (mean 2 958 cm³) than in controls (mean 2 342 cm³). Lung volume ratio values in the drowning group (mean 0.3156) were greater than those in the control group (mean 0.2763); (P = 0.02). There was no significant difference between the drowning and control group in the mean CT value of the whole lung. There were differences between lung CT value distribution curves in drowning victims and controls, with drowning victims showing a single peak and CAD cases showing a bimodal distribution. Thoracic PMCT is helpful for the forensic medical diagnosis of drowning. Lung volume ratio and lung CT value distribution are potential indicators to distinguish between drowning and CAD.     

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.     

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.     

水中尸体肺组织硅藻检验与死因分析

目的 探讨肺组织中硅藻对于判断水中尸体死亡原因的应用价值.方法 收集水中尸体407例,对死亡原因、案件性质、组织器官中硅藻检验结果进行分析.取45只兔按照生前、死后入水及不同季节入水等随机分为9组,应用硝酸消化法处理检材,检测肺组织中硅藻含量.结果 407例水中尸体,硅藻检验阳性372例,其中意外死亡和自杀351例,他...     

Diatom test with Soluene-350 to diagnose drowning in sea water.

The authors tested the Soluene-350 method to detect diatoms in three cases of sea water drowning. The negative results obtained in all three cases prompted us to re-test the method on samples of fresh water and sea water microalgae. The experiment showed that the siliceous frustule of sea water diatoms is solubilized by Soluene-350 while that of fresh water diatoms is resistant to the treatment. The method, which is effective in cases of drowning in fresh water, should therefore be used with caution in sea water drowning.