Fly larvae: a new toxicological method of investigation in forensic medicine

Toxicological analyses on a putrefied cadaver are sometimes difficult to achieve because of the absence of blood and urine. In this study, maggots, living material, are proposed as a new medium of investigation in forensic medicine. Five drugs (triazolam, oxazepam, phenobarbital, alimemazine, and clomipramine) were identified and assayed in some tissues of a putrefied cadaver and in the maggots found on and in the body.     

Fly larvae and their relevance in forensic toxicology

Toxicological analyses on a putrefied cadaver are sometimes difficult to perform because of the absence of blood and urine. In this study, fly larvae, being living material, are proposed as a new medium of investigation in forensic toxicology. Bromazepam and levomepromazine were identified and assayed in the remains of cerebral tissue, in the clavicle of a putrefied cadaver, and in the fly larvae found on and in the corpse.     

Microbial contamination of ABO antigens in bone tissue

Fragments of human bones were stored in different media for two years and then expression of the ABO antigens was indicated. Simultaneously, microbiological investigations were performed. In almost all cases, different ABO substances were detected in putrefied and fresh bones taken from the same person. Blood group antigens found in putrefied bones were compared with serological activity of bacterium cultured from these tissues. Attempts were made to remove unspecific reactions. The authors assume that bacteria are responsible for nonspecific serological reactions, not only as a carrier of blood-group-like substances, but also as a source of enzymes responsible for changes in the structure of ABO antigens in putrefied bones.     

电击兔肢体所致骨损伤后的实验性研究

目的220V交流电击兔肢体后,观察电击部位骨表面的病理改变,为法医病理诊断电击伤提供客观、量化的指标。方法用220V交流电作为损伤电压。动物分为:生前电击组、死后5m in电击组、腐败组及正常对照组。每组骨骼标本均进行肉眼检察、X线检测及SEM观察;同时运用能谱仪测量电击部位元素的种类及含量。结果①电击部位骨皮质SEM下骨皮质紊乱、小孔穴形成、周围高密度影环绕,且钙、磷元素升高,碳元素下降是诊断电击伤的指标,并能鉴别生前、死后电击伤及腐败骨。②小孔穴的形态及钙、磷、碳元素的变化可用于推断电击时间。结论电击造成骨组织的形态及电击部位元素的改变可作为鉴别生前、死后电击伤、腐败骨和推断电击时间的指标。     

Variability of simulants used in recreating stab events

Formic acid (FA) concentration was measured in post-mortem blood and urine samples as methyl formate using a headspace in-tube extraction gas-chromatography-mass-spectrometry method. A total of 113 cases were analyzed, each including a blood and urine sample fortified with 1% sodium fluoride. The cases were divided into three groups: regular (n=59), putrefied (n=30), and methanol-positive (n=22) cases. There was no evidence of ante-mortem methanol consumption in the regular and putrefied cases. In regular cases, the mean (and median) FA concentrations were 0.04 g/l (0.04 g/l) and 0.06 g/l (0.04 g/l) in blood and urine, respectively. In putrefied cases, the mean (and median) FA concentrations were substantially higher, 0.24 g/l (0.22 g/l) and 0.25 g/l (0.15 g/l) in blood and urine, respectively. In three putrefied cases, FA concentration in blood exceeded 0.5 g/l, a level associated with fatal methanol poisoning. Ten putrefied cases were reanalyzed after 3-4 months storage, and no significant changes in FA concentrations were seen. These observations suggest that FA was formed by putrefaction during the post-mortem period, not during sample storage when sodium fluoride was added as a preservative. In methanol-positive cases, the mean (and median) FA concentrations were 0.80 g/l (0.88 g/l) and 3.4 g/l (3.3 g/l) in blood and urine, respectively, and the concentrations ranged from 0.19 to 1.0 g/l in blood and from 1.7 to 5.6 g/l in urine. The mean (and median) methanol concentrations in methanol-positive cases were 3.0 g/l (3.0 g/l) and 4.4 g/l (4.7 g/l) in blood and in urine, respectively. The highest methanol concentrations were 6.0 g/l and 8.7 g/l in blood and urine, respectively. No ethyl alcohol was found in the methanol-positive blood samples. Poor correlation was shown between blood and urine concentrations of FA. Poor correlations were also shown, in both blood and urine, between methanol and FA concentrations.     

Macroscopic, histological and toxicological aspects of early Gammarus pulex scavenging

Two cases of scavenging postmortem freshwater shrimps (Gammarus pulex) are presented. We report the two first illustrated observations of cutaneous postmortem injuries inflicted by a G. pulex population, a small freshwater crustacean, on two non putrefied drowning victims, and we describe their particular histological features and their potential in forensic investigations.     

Detection of marine and freshwater bacterioplankton in immersed victims: Post-mortem bacterial invasion does not readily occur

We previously applied our method of detecting marine or freshwater bacterioplankton (bacteria) in the blood of immersed victims as a marker of drowning. However, we did not confirm the absence of post-mortem bacterial invasion during immersion. Here we examined the nature of bacterioplankton in blood samples from 21 immersed and 4 non-immersed cadavers. We found only freshwater bacterioplankton in the blood of two victims that were retrieved from the sea or an estuary inhabited by marine bacterioplankton even though one victim was highly putrefied. The results of diatom testing suggested that these two victims had drowned in fresh or brackish water with low salinity and then flowed out to the estuary or the sea. Two others were submerged in water, but representative bacterioplankton were undetectable in their blood although one victim was highly putrefied. Autopsy findings and the results of diatom tests did not indicate that the cause of death was drowning. As in previous studies, we identified freshwater bacterioplankton in the blood of seven other victims that had drowned in freshwater, marine bacterioplankton in the blood of four victims that had drowned in seawater and none in four victims found on land that had died by means other than drowning. Bacterioplankton in the blood of drowned victims appears to reflect the type of water aspirated and blood does not become easily contaminated with bacteria post-mortem even in decomposed bodies.     

Infanticide in Malaysia: two case reports and a review of literature

In a strict and conservative society like Malaysia, the number of cases of infanticide has continued to occur. The authors present two typical cases of infanticide in Malaysia. Case 1 concerned a body of a fully mature newborn fetus disposed in a rubbish bin. The head was traumatically amputated by the rubbish truck's compactor. The umbilical cord was still attached to the body, with no reddening around the insertion. The severed neck showed features consistent with post-mortem amputation. The significant finding was expanded crepitant lungs, which floated in water. The histology of the lungs showed expanded alveoli. It was concluded that the baby had been born alive, but no cause of death could be elicited. Case 2 concerned a decomposed mature newborn found in a scrub forest. The internal organs showed advanced putrefaction, the lungs being collapsed, congested, and hemorrhagic, typical of decomposed lungs. No conclusion could be made about the state of birth or the cause of death because of the putrefied state of the body. The two cases illustrate the typical cases and problems faced by pathologists locally and probably elsewhere in the world. Most of the bodies are found in a putrefied state. Pathologists have to ascertain not only maturity and live birth but also the cause of death, which may be very subtle or masked by putrefaction. The problems of diagnosis of live birth are discussed.     

Toxicologic survey of findings in putrefied corpses. Corpse putrefaction in air

Autopsies were conducted on 234 putrefied corpses (exposed to the air, not buried) at the Institute for Forensic Medicine of the University of Hamburg from January 1979 to May 1983. Estimating the time it had taken to reach the respective degree of putrefaction was almost impossible. Only anamnestic clues provided evidence to ascertain the approximate time of death (between 3 days and 46 months ago). The cadavers were found in different locations: outdoors, inside heated or non-heated rooms. Some corpses wore clothes; others were naked; some were covered with blankets or other material. Chemicotoxicological analyses consisted of routine methods and additional modern sophisticated techniques: capillary gas chromatography, combined GC/MS, infrared spectrometry, radio- and enzyme-immunological tests. In organ specimens and body fluids of 47 corpses organic medicaments or poisons could be detected. Nearly all of these 47 forensic autopsies were carried out by request of the legal authorities to rule out legal intoxication as well as the influence of illegal drugs or medicaments. Unlike 20 years ago, today it is possible to detect and quantify organic drugs and poisons as well as their metabolic products, especially minor or major tranquilizers or barbiturates, in highly putrefied specimens. In spite of analytical progress, interpretation of some chemotoxicological results remains difficult. Poor anamnestic data provide insufficient clues. Therefore, pathologicoanatomical and histological results have special significance for the final conclusions. Good cooperation between the pathologist and toxicologist is one of the imperative prerequisites to come to a satisfying result.     

Identification of the human Y-chromosomal microsatellite locus DYS19 from degraded DNA

STR DYS19 seems to be one of the most useful markers for population genetic, evolutionary, and forensic applications. However, the authors have noticed that the amplification of the DYS19 polymorphism fails when highly degraded DNA is used as a template. The authors designed a new pair of primers that reduce the DYS19 fragment sizes compared with those of the known protocol. Using these primers, an improved success rate can be achieved, particularly when putrefied samples are under investigation.     

The determination of carboxyhaemoglobin in the presence of sulphaemoglobin

The quantitative analysis of carboxyhaemoglobin (HbCO) in fresh blood samples containing sulphaemoglobin (SHb) and in post-mortem blood samples was investigated using two automated spectrophotometers. The OSM 3 Hemoximeter gives much more accurate determinations of HbCO than the IL 282 Co-Oximeter in the presence of SHb. In addition, the OSM 3 Hemoximeter is designed to correct for the presence of SHb and turbidity of blood samples. It is thus suitable for the analysis of post-mortem samples from fire victims and decomposed bodies which may be very putrefied and contain SHb.     

Implantation of radio frequency identification device (RFID) microchip in disaster victim identification (DVI)

The tsunami catastrophe of December 2004 left more than 200,000 dead. Disaster victim identification (DVI) teams were presented with the unprecedented challenge of identifying thousands of mostly markedly putrefied and partially skeletised bodies. To this end, an adequate body tagging method is essential. Conventional body bag tagging in terms of writing on body bags and placing of tags inside body bags proved unsatisfactory and problem prone due to consequences of cold storage, formalin (formaldehyde) embalming and body numbers inside storage facilities. The placement of radio frequency identification device (RFID) microchips inside victim bodies provided a practical solution to problems of body tagging and attribution in the DVI setting encountered by the Austrian DVI team in Thailand in early 2005.     

Phosphoglucomutase isoenzymes in human teeth

Powdered dentine samples were prepared from fresh teeth and teeth allowed to putrefy for up to 90 days. In some samples dentine only was included, whereas in others dentine plus pulp remnants were used. Extracts of the powders were tested for phosphoglucomutase activity by starch gel electrophoresis and PGM typing was carried out. Fresh teeth showed activity in at least 50% of cases as did putrefied dentine plus pulp remnants. Putrefied dentine alone showed minimal activity. The method may be useful for identification procedures using fragments of teeth.     

Putrefactive pleural effusions as an alternative sample for drug quantification

In the investigation of drug-related deaths it is occasionally necessary to examine putrefied bodies. In such cases the availability of conventional body fluids for toxicologic analysis is usually limited, whereas blood-stained pleural effusion is often present in adequate quantity in the pleural cavity. Fifty-five cases involving numerous drugs are presented in which pleural effusion drug concentrations are compared with corresponding blood or liver concentrations. More than 90% of comparisons produced coincident interpretations of the role that drugs played in the deaths. In most instances pleural effusion analysis was shown to be a valid alternative to the analysis of blood or other conventional fluids or tissues in cases exhibiting advanced putrefaction.     

Detection of diatoms in blood by a combination of membrane filtering and chemical digestion

An improved method for detecting diatom in blood is reported. Blood of cadavers was obtained by cardiac puncture at inquest or from the left atrium directly at autopsy. The blood was hemolyzed by sodium dodecyl sulfate (SDS) and filtered through membrane filter (47 mm in diameter and 5 microns in pore size). When the blood was putrefied, two or three pieces of membranes filters were needed because of choking membrane pore. The membrane filters were digested with fuming nitric acid and diluted with distilled water. The diluted solution was filtered through membrane filter (25 mm in diameter) again. After drying the membrane filter was immersed in oil. Diatoms on the membrane filter were clearly observed microscopically.     

Time-lapse resistivity surveys over simulated clandestine graves

The aim of this study was to develop a better understanding of how electrical resistivity surveys can be used to locate clandestine graves. Resistivity surveys were conducted regularly over three simulated clandestine graves containing a pig cadaver, no cadaver and a pig cadaver wrapped in tarpaulin, respectively. Additionally, soil and groundwater samples were collected from two more simulated graves outside the survey area. The grave containing a pig cadaver was detectable from a low resistivity anomaly in the survey data. Groundwater data suggest that the resistivity anomaly associated with the surveyed pig grave was caused by a localised increase in groundwater conductivity. Wrapping a cadaver was found to initially change the resistivity response of a grave to a high resistivity anomaly. Resistivity surveys did not detect the disturbed soil in the grave that did not contain a cadaver. Although soil samples showed grave soil to be more porous than undisturbed soil, the lack of response from the grave that did not contain a cadaver suggests that disturbed soil was not responsible for the resistivity anomalies observed in this study. Resistivity surveys successfully detected all graves containing cadavers throughout the study, whilst also showing the potential to eliminate the need for mass excavation in a genuine search.     

Cadaver dog and handler team capabilities in the recovery of buried human remains in the southeastern United States

The detection of human remains that have been deliberately buried to escape detection is a problem for law enforcement. Sometimes the cadaver dog and handler teams are successful, while other times law enforcement and cadaver dog teams are frustrated in their search. Five field trials tested the ability of four cadaver dog and handler teams to detect buried human remains. Human and animal remains were buried in various forested areas during the summer months near Tuscaloosa, Alabama. The remains ranged in decomposition from fresh to skeletonized. Cadaver dogs detected with varying success: buried human remains at different stages of decomposition, buried human remains at different depths, and buried decomposed human and animal remains. The results from these trials showed that some cadaver dogs were able to locate skeletonized remains buried at a significant depth. Fresh and skeletonized remains were found equally by the cadaver dogs along with some caveats. Dog handlers affected the reliability of the cadaver dog results. Observations and videotape of the cadaver dogs during field trials showed that they were reliable in finding buried human remains.     

Detection of methamphetamine and amphetamine in a skeletonized body buried for 5 years

A 28-year-old male methamphetamine abuser, who had been buried for 5 years after being killed by strangulation, was found skeletonized. Methamphetamine and amphetamine in the significantly denatured fatty material of the bone marrow were analyzed by gas chromatography and gas chromatography-mass spectrometry. The confirmation of the chemicals was carried out by chemical ionization (CI) mass chromatography, CI mass spectrometry and CI mass fragmentography. The concentrations of methamphetamine and amphetamine determined by CI mass fragmentography were 1.0 mumol/100 g and 0.1 mumol/100 g, respectively. The method used would seem to be very useful for determination of methamphetamine and amphetamine in marked putrefied biological materials.     

The effects of environment and substrata on deoxyribonucleic acid (DNA): the use of casework samples from New York City

This study was designed to analyze the effects of the environment and substrata on the quality of deoxyribonucleic acid (DNA) isolated from evidentiary specimens. The quality of DNA isolated from actual casework specimens was determined by measuring its size by agarose gel electrophoresis. The information obtained could be used to predict the suitability of the DNA in the samples for restriction fragment length polymorphism (RFLP) analysis. The evidentiary specimens chosen for DNA were classified according to substrate (scrapings, plastic bags, synthetics, denim, and carpet) and according to a subjective evaluation of the condition of the stain (soiled, damp, or putrefied) and to its size (small or large). The results show that DNA of sufficient quality and high molecular weight (HMW) can be reliably isolated from bloodstains deposited on evidentiary items which have an unknown environmental history and which have dried onto a variety of substrata. Subsequent RFLP analysis of a selected number of these samples verified that the DNA was suitable for this type of analysis.