Palaeogenomics of Upper Palaeolithic to Neolithic European hunter-gatherers.

Modern humans have populated Europe for more than 45,000 years1,2. Our knowledge of the genetic relatedness and structure of ancient hunter-gatherers is however limited, owing to the scarceness and poor molecular preservation of human remains from that period3. Here we analyse 356 ancient hunter-gatherer genomes, including new genomic data for 116 individuals from 14 countries in western and central Eurasia, spanning between 35,000 and 5,000 years ago. We identify a genetic ancestry profile in individuals associated with Upper Palaeolithic Gravettian assemblages from western Europe that is distinct from contemporaneous groups related to this archaeological culture in central and southern Europe4, but resembles that of preceding individuals associated with the Aurignacian culture. This ancestry profile survived during the Last Glacial Maximum (25,000 to 19,000 years ago) in human populations from southwestern Europe associated with the Solutrean culture, and with the following Magdalenian culture that re-expanded northeastward after the Last Glacial Maximum. Conversely, we reveal a genetic turnover in southern Europe suggesting a local replacement of human groups around the time of the Last Glacial Maximum, accompanied by a north-to-south dispersal of populations associated with the Epigravettian culture. From at least 14,000 years ago, an ancestry related to this culture spread from the south across the rest of Europe, largely replacing the Magdalenian-associated gene pool. After a period of limited admixture that spanned the beginning of the Mesolithic, we find genetic interactions between western and eastern European hunter-gatherers, who were also characterized by marked differences in phenotypically relevant variants.

Sex estimation of the adult Neandertal Regourdou 1 (Montignac, France): Implications for sexing human fossil remains.

Sex is a biological trait fundamental to the study of hominin fossils. Among the many questions that can be addressed are those related to taxonomy, biological variability, sexual dimorphism, paleoobstetrics, funerary selection, and paleodemography. While new methodologies such as paleogenomics or paleoproteomics can be used to determine sex, they have not been systematically applied to Pleistocene human remains due to their destructive nature. Therefore, we estimated sex from the coxal bone of the newly discovered pelvic remains of the Regourdou 1 Neandertal (Southwest France, MIS 5) based on morphological and metric data employing two methods that have been recently revised and shown to be reliable in multiple studies. Both methods calculate posterior probabilities of the estimate. The right coxal bone of Regourdou 1 was partially reconstructed providing additional traits for sex estimation. These methods were cross validated on 14 sufficiently preserved coxal bones of specimens from the Neandertal lineage. Our results show that the Regourdou 1 individual, whose postcranial skeleton is not robust, is a male, and that previous sex attributions of comparative Neandertal specimens are largely in agreement with those obtained here. Our results encourage additional morphological research of fossil hominins in order to develop a set of methods that are applicable, reliable, and reproducible.

Isotopic calcium biogeochemistry of MIS 5 fossil vertebrate bones: application to the study of the dietary reconstruction of Regourdou 1 Neandertal fossil.

The calcium isotopic composition (δ44/42Ca) of bone and tooth enamel can be used for dietary reconstructions of extant and extinct mammals. In natural conditions, the δ44/42Ca value of bone and teeth varies according to dietary intake with a constant isotopic offset of about -0.6‰. Owing to the poor conservation of collagen, carbon (C), and nitrogen (N) isotopic compositions of the Regourdou Mousterian site (MIS 5, Dordogne, France) previously failed to provide any paleodietary information. Therefore, to reconstruct the trophic chain, we have measured calcium (Ca) isotopes from fossil bone samples of the fauna from the Regourdou site, as well as from three bone samples of the Regourdou 1 Neandertal specimen. The results show a taxon-dependent patterning of the Ca isotopic compositions: herbivores generally have higher δ44/42Ca values than carnivores. All the δ44/42Ca values of Regourdou 1 are low (<-1.6‰), placing this specimen amid carnivores. Using a bone-muscle Ca isotopic offset determined on extant animals, we further show that the δ44/42Ca value of the Regourdou 1 diet, and that of most carnivores, cannot be accounted for by the consumption of meat only, as plants and meat have indistinguishable δ44/42Ca values. Mass balance calculations indicate that the low δ44/42Ca values of the Neandertal's carnivorous diet are explained by the ingestion of bone marrow containing as little as 1% trabecular bone. Our results show that the Regourdou 1 Neanderthal consumed a mixture of various herbivorous prey, as well as trabecular bone, which probably occurred when marrow was ingested, by accident or intentionally.

Cervical ribs in human early life: morphological variability and first identification as a morbidity criterion in a past population.

Despite the medical literature on supernumerary cervical ribs in extant adult samples, little is known about their development and occurrence in early infancy. The documentation of cervical ribs in modern samples of fetuses and neonates is indeed affected by ethical as well as technical limitations. The aim of the present study was to investigate their frequencies and morphological variability in the first known archaeological collection of very young children with this anatomical variant. The study sample comes from the 8B-51 necropolis on the Saï island (Sudan) and dates to the Classic Kerma Period (XVIIIe-XVIe centuries BC). It consists of 64 individuals deceased between 24 weeks of amenorrhoea and 2 years of age. Bilateral or unilateral cervical ribs were found in 27 individuals. A total of 43 cervical ribs were identified, 38 of which are fully preserved. According to these observations, at least 42% of the individuals have unilateral or bilateral cervical ribs, with an average maximum length of < 1 cm. This frequency is very high compared to those observed in contemporary adult samples (up to 3%). First, the comparison of our results with biological and genetic research demonstrating the link between the occurrence of cervical ribs and a reduced chance of survival during infancy allows the first identification of this trait as an indicator of morbidity in an archaeological collection, a morbidity to which a genetic homogeneity or even endogamy could contribute. Second, the number of ribs studied makes it possible to propose a morphological classification based on the general shape and the shape of the articular facets, classification that can be used tos refine the analyses of the trait in future samples.

Cranial trepanation and healing process in modern patients-Bioarchaeological and anthropological implications.

The diagnosis of cranial trepanation in the archaeological record has always been a big challenge for archaeologists. The identification is first and foremost based on the shape and edges of the opening, and in cases where the individual survived, the shape and edges resulted from both the surgical procedure and the healing process. Because of the lack of reliable data on the osseous remodelling process of the skull, it is difficult to distinguish true trepanations from some pathological or physiological defects called pseudotrepanations. Furthermore, it is challenging to investigate the post-operative survival time. The purpose of this paper was to summarize observations made on a total of 90 archaeological samples of cranial lesions interpreted as 'partially' or 'completely' healed trepanations, and compare them with 14 modern case studies. Observations made on monitoring post-operative scans of modern patients, at varying times after craniotomy, provided a preliminary timetable for the successive post-surgery bony changes in the skull, and confirmed that the process of osseous remodelling does lead to smooth and rounded edges of the profile of the opening. However, contrary to what has been observed in several archaeological case studies, none of the cases shows a complete closure of the cranial vault. The sharp vertical edges of the opening become bevelled at late stages of healing, which has to be taken into consideration when interpreting the procedures and methods in past population. By bridging bioarchaeology and medical sciences, this study adds to previous discussions on the diagnosis of healed trepanation in the archaeological record, by providing detailed descriptions of morphological changes at various healing stages, that may help archaeologists to identify ancient trepanations more accurately.

The Neanderthal teeth from Marillac (Charente, Southwestern France): Morphology, comparisons and paleobiology.

Few European sites have yielded human dental remains safely dated to the end of MIS 4/beginning of MIS 3. One of those sites is Marillac (Southwestern France), a collapsed karstic cave where archeological excavations (1967-1980) conducted by B. Vandermeersch unearthed numerous faunal and human remains, as well as a few Mousterian Quina tools. The Marillac sinkhole was occasionally used by humans to process the carcasses of different prey, but there is no evidence for a residential use of the site, nor have any hearths been found. Rare carnivore bones were also discovered, demonstrating that the sinkhole was seasonally used, not only by Neanderthals, but also by predators across several millennia. The lithostratigraphic units containing the human remains were dated to ∼60 kyr. The fossils consisted of numerous fragments of skulls and jaws, isolated teeth and several post-cranial bones, many of them with traces of perimortem manipulations. For those already published, their morphological characteristics and chronostratigraphic context allowed their attribution to Neanderthals. This paper analyzes sixteen unpublished human teeth (fourteen permanent and two deciduous) by investigating the external morphology and metrical variation with respect to other Neanderthal remains and a sample from modern populations. We also investigate their enamel thickness distribution in 2D and 3D, the enamel-dentine junction morphology (using geometric morphometrics) of one molar and two premolars, the roots and the possible expression of taurodontism, as well as pathologies and developmental defects. The anterior tooth use and paramasticatory activities are also discussed. Morphological and structural alterations were found on several teeth, and interpreted in light of human behavior (tooth-pick) and carnivores' actions (partial digestion). The data are interpreted in the context of the available information for the Eurasian Neanderthals.

Issues of theory and method in the analysis of Paleolithic mortuary behavior: A view from Shanidar Cave.

Mortuary behavior (activities concerning dead conspecifics) is one of many traits that were previously widely considered to have been uniquely human, but on which perspectives have changed markedly in recent years. Theoretical approaches to hominin mortuary activity and its evolution have undergone major revision, and advances in diverse archeological and paleoanthropological methods have brought new ways of identifying behaviors such as intentional burial. Despite these advances, debates concerning the nature of hominin mortuary activity, particularly among the Neanderthals, rely heavily on the rereading of old excavations as new finds are relatively rare, limiting the extent to which such debates can benefit from advances in the field. The recent discovery of in situ articulated Neanderthal remains at Shanidar Cave offers a rare opportunity to take full advantage of these methodological and theoretical developments to understand Neanderthal mortuary activity, making a review of these advances relevant and timely.

A case of marked bilateral asymmetry in the sacral alae of the Neandertal specimen Regourdou 1 (Périgord, France).

OBJECTIVES: A marked asymmetry was previously reported in the sacral alae and S1-L5 facets orientation of the Neandertal individual Regourdou 1. Here, we provide a detailed description and quantification of the morphology and degree of asymmetry of this sacrum. MATERIAL AND METHODS: Regourdou 1 was compared to a modern human sample composed of 24 females and 17 males, and to other Neandertal individuals. Both traditional and geometric morphometric analyses were used in order to quantify the degree of sacral asymmetry of Regourdou 1. RESULTS: The asymmetry of both sacral alae and facets orientation substantially exceeds directional and absolute asymmetry of the healthy modern sample. Regourdou 1 shows a considerably shorter right ala, which is absolutely and relatively outside of the modern and Neandertal variations. CONCLUSION: Regourdou 1 shows marked sacral asymmetry that probably originated in early ontogenetic development. An asymmetric sacrum reflects asymmetric load dissipation and could relate to other morphological abnormalities observed in the skeleton, especially the mild scoliosis of the spine and the asymmetry of the femoral diaphyses. Further investigation is necessary to elucidate the relationship between those morphologies as well as a potential impact on the life of the individual.

The costal skeleton of the Regourdou 1 Neandertal.

The morphology and size of the Neandertal thorax is a subject of growing interest due to its link to general aspects of body size and shape, including physiological aspects related to bioenergetics and activity budgets. However, the number of well-preserved adult Neandertal costal remains is still low. The recent finding of new additional costal remains from the Regourdou 1 (R1) skeleton has rendered this skeleton as one of the most complete Neandertal costal skeletons with a minimum of 18 ribs represented, five of which are complete or virtually complete. Here we describe for the first time all the rib remains from R1 and compare them to a large modern Euroamerican male sample as well as to other published Neandertal individuals. The costal skeleton of this individual shows significant metric and morphological differences from our modern human male comparative sample. The perceived differences include: dorsoventrally large 1st and 2nd ribs, 3rd ribs with a very closed dorsal curvature and large maximum diameters at the posterior angle, a large tubercle-iliocostal line distance in the 4th rib, thick shafts at the dorsal end of its 6th ribs, thick mid-shafts of the 8th ribs, large articular tubercles at the 9th ribs, and thick shafts of the 11th and 12th ribs. Here we also describe a new mesosternal fragment: the left lateral half of sternebral segments 4 and 5. This portion reveals that the mesosternum of R1 had a sternal foramen in its inferiormost preserved sternal segment and supports previous estimation of the total length of this mesosternum. The new costal remains from R1 support the view that Neandertals, when compared with modern humans, show a significantly different thorax, consistent with differences found in other anatomical regions such as the vertebral column and pelvis.

Neandertal foot remains from Regourdou 1 (Montignac-sur-Vézère, Dordogne, France).

Regourdou is a well-known Middle Paleolithic site which has yielded the fossil remains of a minimum of two Neandertal individuals. The first individual (Regourdou 1) is represented by a partial skeleton while the second one is represented by a calcaneus. The foot remains of Regourdou 1 have been used in a number of comparative studies, but to date a full description and comparison of all the foot remains from the Regourdou 1 Neandertal, coming from the old excavations and from the recent reanalysis of the faunal remains, does not exist. Here, we describe and comparatively assess the Regourdou 1 tarsals, metatarsals and phalanges. They display traits observed in other Neandertal feet, which are different from some traits of the Sima de los Huesos (Atapuerca) hominins and of Middle Paleolithic, Upper Paleolithic and recent modern humans. These Neandertal features are: a rectangular talar trochlea with a large lateral malleolar facet, a broad talar head, a broad calcaneus with a projecting sustentaculum tali, a wide and wedged navicular with a projecting medial tubercle, large and wide bases of the lateral metatarsals, and mediolaterally expanded and robust phalanges that also show hallux valgus in a strongly built hallux.

Neonatal postcrania from Mezmaiskaya, Russia, and Le Moustier, France, and the development of Neandertal body form.

Neandertal and modern human adults differ in skeletal features of the cranium and postcranium, and it is clear that many of the cranial differences-although not all of them-are already present at the time of birth. We know less, however, about the developmental origins of the postcranial differences. Here, we address this deficiency with morphometric analyses of the postcrania of the two most complete Neandertal neonates-Mezmaiskaya 1 (from Russia) and Le Moustier 2 (from France)-and a recent human sample. We find that neonatal Neandertals already appear to possess the wide body, long pubis, and robust long bones of adult Neandertals. Taken together, current evidence indicates that skeletal differences between Neandertals and modern humans are largely established by the time of birth.

Rabbits in the grave! Consequences of bioturbation on the Neandertal "burial" at Regourdou (Montignac-sur-Vézère, Dordogne).

The understanding of Neanderthal societies, both with regard to their funerary behaviors and their subsistence activities, is hotly debated. Old excavations and a lack of taphonomic context are often factors that limit our ability to address these questions. To better appreciate the exact nature of what is potentially the oldest burial in Western Europe, Regourdou (Montignac-sur-Vézère, Dordogne), and to better understand the taphonomy of this site excavated more than 50 years ago, we report in this contribution a study of the most abundant animals throughout its stratigraphy: the European rabbit (Oryctolagus cuniculus). In addition to questions surrounding the potential bioturbation of the site's stratigraphy, analysis of the Regourdou rabbits could provide new information on Neandertal subsistence behavior. The mortality profile, skeletal-part representation, breakage patterns, surface modification, and comparison with modern reference collections supports the hypothesis that the Regourdou rabbit remains were primarily accumulated due to natural (attritional) mortality. Radiocarbon dates performed directly on the rabbit remains give ages ranging within the second half of Marine Isotope Stage 3, notably younger than the regional Mousterian period. We posit that rabbits dug their burrows within Regourdou's sedimentological filling, likely inhabiting the site after it was filled. The impact of rabbit activity now brings into question both the reliability of the archaeostratigraphy of the site and the paleoenvironmental reconstructions previously proposed for it, and suggests rabbits may have played a role in the distribution of the Neandertal skeletal remains.

Evidence supporting an intentional Neandertal burial at La Chapelle-aux-Saints.

The bouffia Bonneval at La Chapelle-aux-Saints is well known for the discovery of the first secure Neandertal burial in the early 20th century. However, the intentionality of the burial remains an issue of some debate. Here, we present the results of a 12-y fieldwork project, along with a taphonomic analysis of the human remains, designed to assess the funerary context of the La Chapelle-aux-Saints Neandertal. We have established the anthropogenic nature of the burial pit and underlined the taphonomic evidence of a rapid burial of the body. These multiple lines of evidence support the hypothesis of an intentional burial. Finally, the discovery of skeletal elements belonging to the original La Chapelle aux Saints 1 individual, two additional young individuals, and a second adult in the bouffia Bonneval highlights a more complex site-formation history than previously proposed.

Hyperostosis frontalis interna in a Neandertal from Marillac (Charente, France).

The site of Marillac (Charente, France) has yielded an important stratigraphic sequence containing numerous Neandertal remains (some of them with peri-mortem manipulations) from lithofacies 2 (Quina Mousterian). This level has been correlated with MIS 4 and is associated with a TL date of 57,600 ± 4600 years BP (before present). The study of one of the cranial fragments (Marillac 3) revealed a grade 2 or Type B Hyperostosis frontalis interna (HFI), remodelling and altering the internal table of the thick frontal bone. This pathology has been analysed macroscopically together with radiography and sections made using a microscanner and a scanner. The development of the HFI is compared with published evidence for Sangiran 3 (Homo erectus), two other Neandertals (Forbes' Quarry and Shanidar 5), and several archaeological samples. Forbes' Quarry seems to display more advanced HFI than either Shanidar 5 or Marillac 3. The three Neandertals may be considered mature individuals (≥40 years) and it seems likely that the aetiology of this pathology may be associated with hormonal alterations, as has been suggested for past and extant populations. While the prevalence of HFI in contemporary post-menopausal women is well documented, the identification of HFI amongst males from several archaeological samples (Neanderthals, Ancient Egypt, Syrian Bronze Age or the Anasazi), with different stages of development, confirm that the pathology affected both sexes in past populations. Additional data and research are still needed to elucidate the etiopathogenesis of this illness and to better understand the relationship between environmental factors and their possible influences/consequences for the development of metabolic disorders in prehistoric populations.

Neanderthals from El Salt (Alcoy, Spain) in the context of the latest Middle Palaeolithic populations from the southeast of the Iberian Peninsula.

We present a bioanthropological study of dental remains recovered from El Salt Middle Palaeolithic site (Alcoy, Alicante, Spain). The dental remains were found in a sedimentary layer representing a calm depositional environment within a freshwater spring system. The corresponding archaeological context comprises a Middle Palaeolithic faunal and lithic assemblage that represents the last documented evidence of human occupation at the site, dating to between 47.2 ± 4.4 and 45.2 ± 3.4 ka (thousands of years ago). This evidence is overlain by an archaeologically sterile deposit dated to 44.7 ± 3.2 ka. Results show that the teeth belong to a single juvenile or young adult individual with morphological and metric features falling within the Neanderthal range of variability, although the considered traits are not taxonomically highly discriminant. The reported fossils are representative of the latest Middle Palaeolithic groups in the region and may be considered in the ongoing debate on the disappearance of Neanderthals and the end of the Middle Palaeolithic.

Neanderthal infant and adult infracranial remains from Marillac (Charente, France).

At the site of Marillac, near the Ligonne River in Marillac-le-Franc (Charente, France), a remarkable stratigraphic sequence has yielded a wealth of archaeological information, palaeoenvironmental data, as well as faunal and human remains. Marillac must have been a sinkhole used by Neanderthal groups as a hunting camp during MIS 4 (TL date 57,600 ± 4,600BP), where Quina Mousterian lithics and fragmented bones of reindeer predominate. This article describes three infracranial skeleton fragments. Two of them are from adults and consist of the incomplete shafts of a right radius (Marillac 24) and a left fibula (Marillac 26). The third fragment is the diaphysis of the right femur of an immature individual (Marillac 25), the size and shape of which resembles those from Teshik-Tash and could be assigned to a child of a similar age. The three fossils have been compared with the remains of other Neanderthals or anatomically Modern Humans (AMH). Furthermore, the comparison of the infantile femora, Marillac 25 and Teshik-Tash, with the remains of several European children from the early Middle Ages clearly demonstrates the robustness and rounded shape of both Neanderthal diaphyses. Evidence of peri-mortem manipulations have been identified on all three bones, with spiral fractures, percussion pits and, in the case of the radius and femur, unquestionable cutmarks made with flint implements, probably during defleshing. Traces of periostosis appear on the fibula fragment and on the immature femoral diaphysis, although their aetiology remains unknown.

Unraveling the Life History of Past Populations through Hypercementosis: Insights into Cementum Apposition Patterns and Possible Etiologies Using Micro-CT and Confocal Microscopy.

The "teeth-as-tools" hypothesis posits that Neanderthals used their anterior teeth as a tool or a third hand for non-dietary purposes. These non- or para-masticatory activities (e.g., tool-making or food preparation prior to ingestion) have also been described in other past and extant human populations, and other Primates. Cementum is the mineralized tissue that covers the tooth root surface and anchors it to the alveolar bone. Under certain conditions (e.g., mechanical stress, infection), its production becomes excessive (i.e., beyond the physiological state) and is called 'hypercementosis'. Several studies in dental anthropology have established a correlation between the teeth-as-tools and hypercementosis. The present work aims to characterize the different patterns of cementum apposition on archeological teeth and discuss their supposed etiology. Using microtomography and confocal microscopy, the patterns of cementum apposition (i.e., thickness, location, and surface characteristics) were analyzed in 35 hypercementotic teeth (Sains-en-Gohelle, France; 7th-17th c. A.D.). Four groups were identified with distinct hypercementosis patterns: (1) impacted, (2) infected, (3) hypofunctional, and (4) hyperfunctional teeth. Characterizing hypercementosis can contribute to documenting the oral health status (paleopathology) and/or masticatory activity of individuals, even from isolated teeth. This has implications for the study of fossil hominins, particularly Neanderthals, known for their use of anterior teeth as tools and frequent and substantial occurrence of hypercementosis.

The vertebral column of the Regourdou 1 Neandertal.

The Regourdou 1 partial skeleton was found in 1957 in level IV of the eponymous site located in Montignac-sur-Vézère (Dordogne, France) and until now it has been only partially published. The ongoing revision of the faunal remains from the site has yielded additional fossils that pertain to this skeleton. Here we study the vertebral column of this individual, providing for the first time detailed descriptions for all of the fossils and reassessing the anatomical position of all of the fragments. The vertebral column of Regourdou 1 is one of the most complete in the Neandertal fossil record with at least 20 pre-sacral vertebrae (seven cervicals, nine thoracic and four lumbars), a partial sacrum and a fragmentary first coccygeal vertebra. When compared with modern humans, the vertebrae of Regourdou 1 display significant metric differences, and fit well within the range of Neandertal variability. A preliminary analysis of the most complete thoracic vertebrae of this individual indicates that Neandertals displayed significant differences from modern humans in the thoracic spine, which adds to the differences already observed in the cervical and lumbar regions. Finally, we have also observed mild signs of osteoarthrosis, albeit to a lower degree of that present in other Neandertals such as La Chapelle-aux-Saints, La Ferrassie 1 or Shanidar 3. This is consistent with the younger adult age for Regourdou 1.