Scanning Electron Microscopy-Energy-Dispersive X-Ray (SEM/EDX): A Rapid Diagnostic Tool to Aid the Identification of Burnt Bone and Contested Cremains.

This study investigates the use of Scanning electron microscopy-energy-dispersive X-ray (SEM-EDX) as a diagnostic tool for the determination of the osseous origin of samples subjected to different temperatures. Sheep (Ovis aries) ribs of two experimental groups (fleshed and defleshed) were burned at temperatures of between 100°C and 1100°C in 100°C increments and subsequently analyzed with the SEM-EDX to determine the atomic percentage of present elements. Three-factor ANOVA analysis showed that neither the exposure temperature, nor whether the burning occurred with or without soft tissue present had any significant influence on the bone's overall elemental makeup (p > 0.05). The Ca/P ratio remained in the osseous typical range of between 1.6 and 2.58 in all analyzed samples. This demonstrates that even faced with high temperatures, the overall gross elemental content and atomic percentage of elements in bone remain stable, creating a unique "fingerprint" for osseous material, even after exposure to extreme conditions.

The Effect of Soft Tissue on Temperature Estimation from Burnt Bone Using Fourier Transform Infrared Spectroscopy.

This study investigated the effect of soft tissue and different exposure times on the prediction of burning temperatures of bone when using Fourier transform infrared spectroscopy (FTIR). Ovis aries rib bones were burnt at different temperatures and for varying time intervals. Results of a linear regression analysis indicated that burn temperatures can be predicted with a standard error of ±70 °C from defleshed bone spectra. Exposure time does not have a significant impact on prediction accuracy. The presence of soft tissue has a significant impact on heat-induced changes of the bone matrix in low (<300 °C) as well as high temperatures (>800 °C), slowing down combustion in the former and accelerating it in the latter (p < 0.05). At medium temperatures, no significant difference was noted. These results provide forensic investigators a new perspective with which to interpret the results of crystallinity measures derived from burnt bone.

Soil fungal community shift evaluation as a potential cadaver decomposition indicator.

Fungi metabolise organic matter in situ and so alter both the bio-/physico-chemical properties and microbial community structure of the ecosystem. In particular, they are responsible reportedly for specific stages of decomposition. Therefore, this study aimed to extend previous bacteria-based forensic ecogenomics research by investigating soil fungal community and cadaver decomposition interactions in microcosms with garden soil (20 kg, fresh weight) and domestic pig (Sus scrofa domesticus) carcass (5 kg, leg). Soil samples were collected at depths of 0-10 cm, 10-20 cm and 20-30 cm on days 3, 28 and 77 in the absence (control -Pg) and presence (experimental +Pg) of Sus scrofa domesticus and used for total DNA extraction and nested polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) profiling of the 18S rRNA gene. The Shannon-Wiener (H') community diversity indices were 1.25±0.21 and 1.49±0.30 for the control and experimental microcosms, respectively, while comparable Simpson species dominance (S) values were 0.65±0.109 and 0.75±0.015. Generally, and in contrast to parallel studies of the bacterial 16S rRNA and 16S rDNA profiles, statistical analysis (t-test) of the 18S dynamics showed no mathematically significant shifts in fungal community diversity (H'; p=0.142) and dominance (S; p=0.392) during carcass decomposition, necessitating further investigations.

Estimating temperature exposure of burnt bone - A methodological review.

Forensic anthropologists are frequently confronted with the need to interpret burnt bone. Regardless of the context, one of the key factors for the correct interpretation of the remains and a reconstruction of the incidents leading to incineration is the estimation of the maximum exposure temperature. The recent years have seen an influx in experimental research focusing on temperature estimation, spanning from colour assessment, mechanical strength measurements, histology and structural observations, biochemical changes and crystallinity studies, vastly advancing the understanding of heat induced changes in bone, thus facilitating a more accurate interpretation. This paper draws together and evaluates all currently available methodologies for temperature estimation.

Sexual dimorphism of the lateral angle of the internal auditory canal and its potential for sex estimation of burned human skeletal remains.

The potential of the petrous bone for sex estimation has been recurrently investigated in the past because it is very resilient and therefore tends to preserve rather well. The sexual dimorphism of the lateral angle of the internal auditory canal was investigated in two samples of cremated Portuguese individuals in order to assess its usefulness for sex estimation in burned remains. These comprised the cremated petrous bones from fleshed cadavers (N = 54) and from dry and disarticulated bones (N = 36). Although differences between males and females were more patent in the sample of skeletons, none presented a very significant sexual dimorphism, thus precluding any attempt of sex estimation. This may have been the result of a difficult application of the method and of a differential impact of heat-induced warping which is known to be less frequent in cremains from dry skeletons. Results suggest that the lateral angle method cannot be applied to burned human skeletal remains.

An investigation into the persistence of textile fibres on buried carcasses.

A significant amount of research has been carried out on fibres to aid the forensic scientist in determining the significance of these when found on a victim or suspect. This work has focused on open-air environments, and as such no research has been undertaken to examine the persistence of fibres on bodies in the burial environment. Wool and cotton fibres, known to fluoresce under ultraviolet (UV) light, were transferred onto the skin of four porcine (Sus scrofa) carcasses (two carcasses per fibre type). The number of fibres transferred was recorded from images taken under UV light. The remains were subsequently placed in four burial sites and left interred for 14 days. After this period the carcasses were excavated and lightly brushed down to remove the soil layer that had adhered to the skin. Once again photography under UV light was used to record the number of fibres which persisted on the skin. Results showed that after 14 days, wool and cotton fibres remain on the surface of the buried carcasses. In no circumstance was there a total loss of fibres suggesting that in such scenarios, the likelihood of finding fibres is high but the initial number of fibres transferred would be strongly diminished. This has important implications for both the excavation protocol for buried remains and the subsequent analysis for physical evidence.

Weight references for burned human skeletal remains from Portuguese samples.

Weight is often one of the few recoverable data when analyzing human cremains but references are still rare, especially for European populations. Mean weights for skeletal remains were thus documented for Portuguese modern cremations of both recently deceased individuals and dry skeletons, and the effect of age, sex, and the intensity of combustion was investigated using both multivariate and univariate statistics. The cremains from fresh cadavers were significantly heavier than the ones from dry skeletons regardless of sex and age cohort (p < 0.001 to p = 0.003). As expected, males were heavier than females and age had a powerful effect in female skeletal weight. The effect of the intensity of combustion in cremains weight was unclear. These weight references may, in some cases, help estimating the minimum number of individuals, the completeness of the skeletal assemblage, and the sex of an unknown individual.

Investigation into the merits of infrared imaging in the investigation of tattoos postmortem.

Infrared imaging has a history of use in the forensic examination of artwork and documents and is investigated here for its wider use in the detection of tattoos on the human body postmortem. Infrared photographic and reflectographic techniques were tested on 18 living individuals, displaying a total of 30 tattoos. It was observed that neither age, sex, age of the tattoo, nor, most significantly, skin color affected the ability to image the tattoos using infrared imaging techniques. Second, a piglet carcass was tattooed and the impact of the decomposition process on the visibility of the tattoos assessed. Changes were recorded for 17 days and decomposition included partial mummification and skin discoloration. Crucially, the discoloration was recorded as greatly affecting the image quality using conventional photography, but was insignificant to the infrared recording of these tattoos. It was concluded that infrared reflectography was beneficial in the investigation into tattoos postmortem.

The potential of X-ray diffraction in the analysis of burned remains from forensic contexts.

In view of the difficulties in extracting quantitative information from burned bone, we suggest a new and accurate method of determining the temperature and duration of burning of human remains in forensic contexts. Application of the powder X-ray diffraction approach to a sample of human bone and teeth allowed their microstructural behavior, as a function of temperature (200-1000 degrees C) and duration of burning (0, 18, 36, and 60 min), to be predicted. The experimental results from the 57 human bone sections and 12 molar teeth determined that the growth of hydroxylapatite crystallites is a direct and predictable function of the applied temperature, which follows a nonlinear logistic relationship. This will allow the forensic investigator to acquire useful information about the equilibrium temperature brought about by the burning process and to suggest a reasonable duration of fire exposure.