Burnt Bone Fragments Research Articles

Robust High-Throughput Proteomics Identification and Deamidation Quantitation of Extinct Species up to Pleistocene with Ultrahigh-Resolution MALDI-FTICR Mass Spectrometry.

Peptide mass fingerprinting (PMF) using MALDI-TOF mass spectrometry allows the identification of bone species based on their type I collagen sequence. In the archaeological or paleontological field, PMF is known as zooarchaeology mass spectrometry (ZooMS) and is widely implemented to find markers for most species, including the extinct ones. In addition to the identification of bone species, ZooMS enables dating estimation by measuring the deamidation value of specific peptides. Herein, we report several enhancements to the classical ZooMS technique, which reduces to 10-fold the required bone sample amount (down to the milligram scale) and achieves robust deamidation value calculation in a high-throughput manner. These improvements rely on a 96-well plate samples preparation, a careful optimization of collagen extraction and digestion to avoid spurious post-translational modification production, and PMF at high resolution using matrix-assisted laser desorption ionization Fourier transform ion cyclotron resonance (MALDI-FTICR) analysis. This method was applied to the identification of a hundred bones of herbivores from the Middle Paleolithic site of Caours (Somme, France) well dated from the Eemian Last Interglacial climatic optimum. The method gave reliable species identification to bones already identified by their osteomorphology, as well as to more challenging samples consisting of small or burned bone fragments. Deamidation values of bones originating from the same geological layers have a low standard deviation. The method can be applied to archaeological bone remains and offers a robust capacity to identify traditionally unidentifiable bone fragments, thus increasing the number of identified specimens and providing invaluable information in specific contexts.

Survival of sharp force trauma in burnt bones: effects of environmental factors

This study investigates how environmental variables, such as temperature and rainfall, affect previously induced cut marks on burnt bones. This research used non-serrated and serrated blade knives to inflict trauma on Sus scrofa ribs (n = 240). The bones were later burnt and left for 1 month in a taphonomic experimental facility. Qualitative and quantitative examinations were conducted using macroscopic and microscopic techniques to assess specific characteristics of the cut marks. Any changes to the dimension and morphology of the cut marks as well as their level of fragmentation were recorded.This study has led to three important outcomes: (1) identification of pre-existing cut marks is possible in reconstructed burnt bone fragments; (2) cut marks from different types of knife blades showed dissimilar responses to heat and the environment; and (3) specific environmental variables affect burnt bone fragmentation. These results have implications for trauma analysis on burnt remains in forensic anthropology casework.

Salorno—Dos de la Forca (Adige Valley, Northern Italy): A unique cremation site of the Late Bronze Age

The archaeological site of Salorno-Dos de la Forca (Bozen, Alto Adige) provides one of the rarest and most significant documentations of cremated human remains preserved from an ancient cremation platform (ustrinum). The pyre area, located along the upper Adige valley, is dated to the Late Bronze Age (ca. 1150-950 BCE) and has yielded an unprecedented quantity of cremated human remains (about 63.5 kg), along with burnt animal bone fragments, shards of pottery, and other grave goods made in bronze and animal bone/antler. This study focuses on the bioanthropological analysis of the human remains and discusses the formation of the unusual burnt deposits at Salorno through comparisons with modern practices and protohistoric and contemporaneous archaeological deposits. The patterning of bone fragmentation and commingling was investigated using spatial data recorded during excavation which, along with the bioanthropological and archaeological data, are used to model and test two hypotheses: Salorno-Dos de la Forca would be the result of A) repeated primary cremations left in situ; or B) of residual material remaining after select elements were removed for internment in urns or burials to unknown depositional sites. By modelling bone weight and demographic data borrowed from regional affine contexts, the authors suggest that this cremation site may have been used over several generations by a small community-perhaps a local elite. With a quantity of human remains that exceeds that of any other coeval contexts interpreted as ustrina, Salorno may be the product of a complex series of rituals in which the human cremains did not receive individual burial, but were left in situ, in a collective/communal place of primary combustion, defining an area of repeated funeral ceremonies involving offerings and libations across a few generations. This would represent a new typological and functional category that adds to the variability of mortuary customs at the end of the Bronze Age in the Alpine are, at a time in which "globalising" social trends may have stimulated the definition of more private identities.

Knife cut marks inflicted by different blade types and the changes induced by heat: a dimensional and morphological study

Detailed information on skeletal trauma analysis of burned bone is important to ascertain the manner and cause of death in forensic casework. This research used three different knife types, one with a non-serrated blade, one a fine-serrated blade, and one a coarse-serrated blade, to inflict trauma to manually macerated Sus scrofa ribs (n = 240), and these ribs were later exposed to heat. Qualitative and quantitative analyses were conducted using macroscopic and microscopic techniques to assess specific characteristics of the cut marks. Differences in cut mark dimension and morphology of the ribs were investigated. After heat exposure, the cut marks on the rib samples remained recognisable and did not alter considerably. A level of dimensional and morphological preservation was reliant on the cutting action and the features of the knife blade as well as surrounding bone injury. The cut marks inflicted by the non-serrated blade remained recognisable despite exposure to the burning process. However, the cut marks inflicted by the coarse-serrated blade were likely to change significantly when exposed to heat. This study leads to two important results: (1) identification of pre-existing cut marks prior to heat exposure is possible in reconstructed burned bone fragments, and (2) cut marks from different types of knife blades showed dissimilar responses to heat. The outcomes obtained in this study stressed the need to adopt great care with the effects of heat on skeletal trauma analysis.

Who was buried with Nestor's Cup? Macroscopic and microscopic analyses of the cremated remains from Tomb 168 (second half of the 8th century BCE, Pithekoussai, Ischia Island, Italy).

Cremation 168 from the second half of the 8th century BCE (Pithekoussai’s necropolis, Ischia Island, Italy), better known as the Tomb of Nestor’s Cup, is widely considered as one of the most intriguing discoveries in the Mediterranean Pre-Classic archaeology. A drinking cup, from which the Tomb’s name derives, bears one of the earliest surviving examples of written Greek, representing the oldest Homeric poetry ever recovered. According to previous osteological analyses, the Cup is associated with the cremated remains of a juvenile, aged approximately 10–14 years at death. Since then, a vast body of literature has attempted to explain the unique association between the exceptionality of the grave good complex, the symposiac and erotic evocation of the Nestor’s Cup inscription with the young age of the individual buried with it. This paper reconsiders previous assessments of the remains by combining gross morphology with qualitative histology and histomorphometric analyses of the burnt bone fragments. This work reveals the commingled nature of the bone assemblage, identifying for the first time, more than one human individual mixed with faunal remains. These outcomes dramatically change previous reconstructions of the cremation deposit, rewriting the answer to the question: who was buried with Nestor’s Cup?.

THE CORE OF WAJO CONFEDERATION: ARCHAEOLOGICAL SURVEY IN TOSORA, CINNOTABI AND LAMASEWANUA

“The beginning of Wajo's establishment began with the migration of people from various places to open rice fields and build settlements to the east of Lake Tempe. The settlements were then transformed into political units which formed a 'state' under the rule of a nobleman based in Cinnotabi. Several political agreements underlie the formation of a confederation of three domains. Regime change resulted in the transfer of the center of government. Lontara Wajo and the oral tradition mention some toponyms but do not explain in detail where the core of the Wajo confederation lies. This research is aimed at determining the location and character of the toponym by conducting field surveys in places that are suspected of being associated with the existence of the pre-Islamic capital of Wajo. Using an archaeological approach and supported by information from textual sources. Surveys in the villages of Tosora, Cinnongtabi and Tajo in Majauleng District have identified the existence of the old capitals around Wajo-wajo, Boli, Leppadeppa, Attunuang, and other sites based on archeological traces such as menhirs, burned bone fragments, pottery and ceramics shards and other artifacts. Identification of imported ceramic fragments from China, Thailand, Vietnam, these sites might be dated between the 14th and 17th centuries. Taking into account the concentration of artefacts and relations between sites, it can be concluded that Tosora was the capital from the end of the 16th century and until the arrival of Islam at the beginning of the 17th century, while the capital of the early period of Wajo hypothetically was dated between the beginning of the 15th century and the end of the 16th century was around the confluence of Wajo-wajoe river which flows into Latamperu and Penrange lake which then empties into Cellue river before ending at the Cenrana mainstream”

The molecular-genetic analysis of mitochondrial DNA from the burnt bones: 'the limits of the possible' problem revisited

The authors report the results of the demonstrative study continuing the cycle of interactive discussions pertinent to the possibility of obtaining reliable genetic information from the analysis of burnt bone fragments. Special emphasis is placed on the worthiness of these materials for genotyping of mitochondrial DNA (mtDNA) with the use of the standard analytical methods employed for the purpose of forensic medical expertise to investigate into the length polymorphism of the amplified mtDNA fragments (PAF) by means of sequencing with fluorescent detection. The study has demonstrated that the mtDNA fragments in the state suitable for genotyping can be found only in the preparations from the bone tissue exposed to the 'mild' thermal impact after which the affected bone is virtually indistinguishable from the native one as far as the outward appearance is concerned. In the cases of a more rigorous thermal impact when the bone tissue exhibits well pronounced signs of heat destruction, it should be considered as inherently unsuitable for genotyping of mtDNA. It was shown that chromosomal DNA is inferior to mtDNA in terms of heat resistance. This finding agrees with the currently adopted view, however this advantage of mtDNA is relatively insignificant from the standpoint of genotyping efficiency.

Scientific evidence for the identification of an Aboriginal massacre at the Sturt Creek sites on the Kimberley frontier of north-western Australia

Archival research into episodes of frontier violence in the Kimberley region of Western Australia indicate that the bodies of Aboriginal victims of massacres were frequently incinerated following the event. This paper presents the results of a scientific investigation of a reported massacre at Sturt Creek where burnt bone fragments were identified in two adjacent sites and documents the archaeological signatures associated with the sites. The methodology used to undertake the project brought together three systems of knowledge: the oral testimonies of the descent group originating from a sole adult survivor of the massacre; archival, historical and scientific research. An archaeological survey defined the two distinct sites containing hundreds of fragile bone fragments; a third site was found to be highly disturbed. Scientific investigations included macroscopic and microscopic examination of selected bone fragments by an anatomical pathologist and a zooarchaeologist and X-ray diffraction analysis of sixteen bone fragments.The anatomical pathologist and zooarchaeologist undertook macroscopic and microscopic examinations of selected bone samples to identify morphological evidence for human origin. It was concluded that three bone fragments examined may have been human, and two of the fragments may have been from the vault of a skull. It was concluded that the likelihood of them being human would be strengthened if it was found that the three samples had been subjected to high temperatures. X-ray diffraction analysis of 16 bone fragments provided this evidence. All fragments showed sharp hydroxylapatite peaks (crystallite sizes 9882nm and 597nm respectively) and all had been subjected to extreme temperatures of either 600°C for more than 80h, 650°C for more than 20h, 700°C for more than 4h or 800°C for more than 1h. XRD analyses were also done on bone samples collected from three cooking hearths at three different archaeological sites. It was found that two of the three samples had been exposed to substantially lower temperatures for a short time period. It was concluded that there was strong pathological and archaeological evidence that the bone fragments were human in origin, but that the evidence was not conclusive. This research also identified archaeological signatures for the identification of massacre sites in similar Australian environments and circumstances.

The Saga of Winkler-1: A Midland Site in Southeast New Mexico

ABSTRACTWinkler-1 is a unique late Pleistocene campsite. The site deposit was found eroding from a buried sandy matrix in Lea County, New Mexico. It was systematically mapped and collected over a period of more than two decades. Chipped stone tools, lithic debris, large mammal bone fragments, and an eyed bone needle comprise the artifact assemblage. Forty-eight Midland point bases, reworked points, and fragments were recovered from the eroding surfaces throughout the site. Edwards chert makes up approximately 85 per cent of the chipped stone assemblage. Three different radiocarbon labs have determined that burned bone fragments date from the mid-10,000s to the mid-9000s 14C yr BP. Unlike other sites where Midland and Folsom artifacts are found mixed together or barely superimposed, Winkler-1 is the only known pure Midland site assemblage that has been thoroughly studied and reported.

Exploring Integrated Geophysics and Geotechnics as a Paleolandscape Reconstruction Tool: Archaeological Prospection of (Prehistoric) Sites Buried Deeply below the Scheldt Polders (NW Belgium)

AbstractDuring extensive construction works in Antwerp harbour, well preserved Late Glacial dune formations were discovered buried deeply below the Scheldt polders and covered by peat, organic matter (OM) rich clays and marine clayey to sandy sediments. First, coring based archaeological prospection strategies for evaluating prehistoric occupation levels in wetland landscapes are reviewed. Next, a more effective approach including near surface geophysical and geotechnical techniques is proposed and tested in Doelpolder Noord. The results indicate that high resolution electromagnetic induction survey at multiple coil spacing provides a suitable approximation of the prehistoric landscape variability but is challenged by variations in groundwater brackishness. Gridded cone penetration tests provide a solution in such cases and serve as an excellent interpretation tool for the conductivity data in general. Due to the required effort, electrical resistance imaging and shear wave land seismics were judged inefficient. Finally, a small dune with indications of paleosol conservation and estimated suitability for Final Palaeolithic to Early Neolithic occupation is sampled by Dutch hand augering and Sonic Drill Aqualock coring. Archaeological indicators for prehistoric occupation such as burnt bone and flint fragments were retrieved from these samples after sieving. Copyright © 2016 John Wiley & Sons, Ltd.

A new Cambrian black pigment used during the late Middle Palaeolithic discovered at Scladina Cave (Andenne, Belgium)

Sedimentary Unit 1A at Scladina Cave, Belgium has yielded archaeological material from a Middle Palaeolithic occupation dating to between 40,210 + 400/−350 BP and 37,300 + 370/−320 BP. Fifty-one fragments of a black, friable rock with a black streak were found in association with 194 burned bone fragments and several thousand lithic artefacts. This black material is interpreted as a pigment brought to the site by Neandertals.The pigment was analysed by petrography, XRD, Raman microspectroscopy, and other geochemical methods. It was identified as a highly siliceous graphitic siltstone. This is a very unique discovery, as European archaeological research has so far only recorded black pigments comprised of manganese oxides from the Middle Palaeolithic. Raman microspectroscopy is a non-destructive method able to distinguish the attributes of black siliceous materials that originate from different tectono-sedimentary contexts. By measuring the degree of alteration of the carbonaceous material, this method allowed for the determination of its geographical and geological origins: a Cambrian formation of very limited extent located near Ottignies, about 40 kilometres north-west of Scladina Cave. The absence of a drainage network connecting the two locations eliminates the possibility of natural transport, and supports its anthropogenic origin.

DNA survival and physical and histological properties of heat-induced alterations in burnt bones

During forensic casework, it is vital to be able to obtain valuable information from burnt bone fragments to ascertain the identity of the victim. Here, we report the findings of an experimental study on burnt bovine compact bone segments. Compact bones were cut to size and heated in an electric furnace at a temperature range of 100–1,100 °C with 100 °C increments. Heat-induced alterations to the bone color,weight, volume, and density were monitored using gross morphology and micro-focus X-ray computed tomography.We found that the increase in temperature caused the color of the compact bones to change in order of yellow, brown, gray,and white. In contrast to the weight reduction that occurred immediately after burning, we measured no significant reduction in volume even at 600 °C; however, volume reduced drastically once the temperature reached 700 °C. Light microscopic histological observations of burnt bone revealed heat induced alterations such as cracking and separation of the osteons at higher temperatures. In addition to these findings,we sought to examine the survival of DNA in the burnt bones using polymerase chain reaction of mitochondrial DNA. No amplification was found in the specimens burnt at 250 °C or higher, indicating the likely difficulty in testing the DNA of burnt bones from forensic casework. The results of this study will enable an estimation of the burning temperatures of burnt bones found in forensic cases and will provide an important framework with which to interpret data obtained during anthropological testing and DNA typing.

River floodplain aggradation history and cultural activities: Geoarchaeological investigation at the Yuezhuang site, Lower Yellow River, China

Archaeobotanical research at the Yuezhuang and Xihe sites of the Houli culture in the Lower Yellow River has found abundant carbonised plant remains, including rice, millet, weed seeds and many other plants (7800–7000 cal BP). Although studies of the diverse economic practices of the Houli people have begun, issues such as ecological diversity and relationships between site formation processes with environmental changes are still poorly understood. Applying soil micromorphology and related methods such as particle size distribution and loss-on-ignition at Yuezhuang, this paper presents results of the geoarchaeological investigation and the implications for the reconstruction of site formation processes in relation to river alluvial history. A long alluvial sequence is reconstructed, consisting of frequent alternations of short-periods surface stabilities and succeeding alluviations. Human occupations corresponded to those short-term surface stabilities and are evidenced by the presence of anthropogenic inclusions in thin sections, including burned bone fragments and pottery sherds. The significances of this study to an ecological approach focusing on environmental changes and cultural adaption are discussed.

Neolithic cremation in south‐east Arabia: archaeological and anthropological observations at FAY‐NE10 in the Emirate of Sharjah (UAE)

Small fragments of cremated human bone, clearly representing numerous individuals, were found in a stratified Neolithic context in a small cave at Jebel Faya in the Central Region of the Emirate of Sharjah, United Arab Emirates. With radiocarbon dates between 6500 and 5800 cal BC, they are among the earliest well‐documented cases of cremations in south‐west Asia. Taxonomic identification of the small burned bone fragments is based on morphological and histological analysis. The deposits are to date the oldest human remains found in south‐east Arabia and represent a phase during the Middle Neolithic for which no other types of burials have yet been documented.

Elemental analysis of burnt human bone for classifying sex and age at death by logistic regression

This study aims to develop a protocol for classifying sex and age groups of deceased persons by using the chemical information in burnt bone fragments. Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES) has been employed to assess the chemical compositions of the burnt bones from the deceased, all of Thai nationality. By performing three ICP-OES measurements on burnt bone samples from 75 individuals, 43 males and 32 females, with an accurate record of their age at death, only six elements i.e. Al, Ca, K, Mg, P, S, were observed in the fragments above the instrumental detection limit of ICP-OES. Fifteen concentration ratios of these particular elements were examined for their significant difference with respect to sex and age group by Mann–Whitney U and Kruskal–Wallis tests, respectively. Subsequently, the significantly different ratios were employed to assess the relationship between bone chemical compositions and sex/age groups of the deceased using logistic regression analysis. Binary and multinomial logistic regressions were required to classify sex and age group at death, respectively. Regression analyses indicated that the input variables representing sex should be included for classifying the age group of the deceased. The chemical information in burnt bone fragments could be employed to assess the sex and age groups of the deceased with 79.60% and 75.10% accuracy, respectively. Chemical measurements were performed on burnt bone samples from two additional individuals to verify the developed logistic equations and classification protocol. More fractions of bones from an individual give a higher percentage of correction for classifying sex and age at death of the deceased.

Vikingetidens gravskik i Danmark – Spor af begravelsesritualer i jordfæstegrave

Vikingetidens gravskik i Danmark – Spor af begravelsesritualer i jordfæstegrave

Reliable genetic identification of burnt human remains

The identification of severely burnt human remains by genetic fingerprinting is a common task in forensic routine work. In cases of extreme fire impact, only hard tissues (bones, teeth) may be left for DNA analysis. DNA extracted from burnt bone fragments may be highly degraded, making an amplification of genetic markers difficult or even impossible. Furthermore, heavily burnt bones are very prone to contamination with external DNA. We investigated whether authentic DNA profiles can be generated from human bones showing different stages of fire induced destruction (well preserved, semi-burnt, black burnt, blue–grey burnt, blue–grey–white burnt). DNA was extracted from 71 bone fragments derived from 13 individuals. Obtained genetic patterns (STRs and mtDNA sequences) were compared to the genetic pattern of the respective bodies. Our results show that the identification via DNA analysis is reliably and reproducibly possible from well preserved and semi-burnt bones. In black burnt bones the DNA was highly degraded and in some cases no nuclear DNA was left, leaving mitochondrial DNA analysis as an option. Blue–grey burnt bones lead only sporadically to authentic profiles. The investigation of blue–grey–white burnt bones barely led to reliable results.

A study by thermal treatment and X-ray powder diffraction on burnt fragmented bones from tombs II, IV and IX belonging to the hypogeic necropolis of “Sa Figu” near Ittiri, Sassari (Sardinia, Italy)

In the course of an anthropological investigation of three tombs of the hypogeic necropolis of “Sa Figu” (near the village of Ittiri in North Sardinia, Italy), numerous burnt bone fragments were collected. In particular, from the whole necropolis we selected a tenth of long bones from tomb IV, a small fragment of femur from tomb II and four fragments belonging to tomb IX. The aim of this work is to determine a temperature range to which the bone fragments were heated, in order to assess a funerary cremation which was presumably used at that time to quickly and hygienically eliminate the flashy parts of deceased people rather than a purification rite with fires inside the sepulchres. We attempted to evaluate the range of temperature to which the bones were likely subjected, making a joint use of thermal treatment and powder X-ray diffraction investigations. From our X-ray line broadening results carried out with a modern approach (Rietveld method), it emerged that a group of five fragmented burnt bones (one specimen belonging to tomb IV, the other four from tomb IX) were subjected to a heat treatment in a temperature range from 600 and 750 °C. Conversely another group of bones (belonging to tombs II and IV) turned out to be subjected to temperatures not higher than 250 °C.

A Novel Model to Test Accuracy and Reproducibility of MDCT Scan Protocols for Coronary Calcium in Vivo

We compared the accuracy and reliability of prospectively triggered, retrospectively ECG gated, and non-gated CT image reconstruction for measurements of coronary artery calcification (CAC) in vivo using a novel animal model. In six Yorkshire farm pigs, prefabricated chains of cortical bone fragments were sutured over the epicardial bed of the major coronary arteries. Using a 4-slice MDCT scanner, each animal was imaged with two different protocols: sequential acquisition with prospective ECG triggering, and spiral acquisition with retrospectively ECG gated image reconstruction- non-gated reconstructions were also generated from these latter scans. Two independent observers measured the 'Agatston score' (AS), the calcified volume (CV), and mineral mass (MM). To calculate accuracy of MM measurements the ash weight of the burned bone fragments was compared to MDCT derived MM. Six pigs successfully underwent surgery and CT imaging (mean heart rate: 86+/-12 bpm). MM measurements from prospectively ECG triggered CT sequential scans were more accurate (p<0.02) and reproducible (p=0.05) than sequential CT scans without ECG triggering or spiral acquisition using retrospective ECG gating. At high heart rates prospective ECG triggered image reconstruction is more accurate and reproducible for CAC scoring than retrospective ECG gated reconstruction and non-gated reconstruction.

Correlation between the responses of lymphocytes and unpleasant emotional behaviour elicited by hypothalamic stimulation in cats

In the course of an anthropological investigation of three tombs of the hypogeic necropolis of “Sa Figu” (near the village of Ittiri in North Sardinia, Italy), numerous burnt bone fragments were collected. In particular, from the whole necropolis we selected a tenth of long bones from tomb IV, a small fragment of femur from tomb II and four fragments belonging to tomb IX. The aim of this work is to determine a temperature range to which the bone fragments were heated, in order to assess a funerary cremation which was presumably used at that time to quickly and hygienically eliminate the flashy parts of deceased people rather than a purification rite with fires inside the sepulchres. We attempted to evaluate the range of temperature to which the bones were likely subjected, making a joint use of thermal treatment and powder X-ray diffraction investigations. From our X-ray line broadening results carried out with a modern approach (Rietveld method), it emerged that a group of five fragmented burnt bones (one specimen belonging to tomb IV, the other four from tomb IX) were subjected to a heat treatment in a temperature range from 600 and 750 °C. Conversely another group of bones (belonging to tombs II and IV) turned out to be subjected to temperatures not higher than 250 °C.