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      A Community in Life and Death: The Late Neolithic Megalithic Tomb at Alto de Reinoso (Burgos, Spain)

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          Abstract

          The analysis of the human remains from the megalithic tomb at Alto de Reinoso represents the widest integrative study of a Neolithic collective burial in Spain. Combining archaeology, osteology, molecular genetics and stable isotope analysis ( 87Sr/ 86Sr, δ 15N, δ 13C) it provides a wealth of information on the minimum number of individuals, age, sex, body height, pathologies, mitochondrial DNA profiles, kinship relations, mobility, and diet. The grave was in use for approximately one hundred years around 3700 cal BC, thus dating from the Late Neolithic of the Iberian chronology. At the bottom of the collective tomb, six complete and six partial skeletons lay in anatomically correct positions. Above them, further bodies represented a subsequent and different use of the tomb, with almost all of the skeletons exhibiting signs of manipulation such as missing skeletal parts, especially skulls. The megalithic monument comprised at least 47 individuals, including males, females, and subadults, although children aged 0–6 years were underrepresented. The skeletal remains exhibited a moderate number of pathologies, such as degenerative joint diseases, healed fractures, cranial trauma, and a low intensity of caries. The mitochondrial DNA profiles revealed a pattern pointing to a closely related local community with matrilineal kinship patterns. In some cases adjacent individuals in the bottom layer showed familial relationships. According to their strontium isotope ratios, only a few individuals were likely to have spent their early childhood in a different geological environment, whilst the majority of individuals grew up locally. Carbon and nitrogen isotope analysis, which was undertaken to reconstruct the dietary habits, indicated that this was a homogeneous group with egalitarian access to food. Cereals and small ruminants were the principal sources of nutrition. These data fit in well with a lifestyle typical of sedentary farming populations in the Spanish Meseta during this period of the Neolithic.

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          Ancient human genomes suggest three ancestral populations for present-day Europeans

          We sequenced genomes from a $\sim$7,000 year old early farmer from Stuttgart in Germany, an $\sim$8,000 year old hunter-gatherer from Luxembourg, and seven $\sim$8,000 year old hunter-gatherers from southern Sweden. We analyzed these data together with other ancient genomes and 2,345 contemporary humans to show that the great majority of present-day Europeans derive from at least three highly differentiated populations: West European Hunter-Gatherers (WHG), who contributed ancestry to all Europeans but not to Near Easterners; Ancient North Eurasians (ANE), who were most closely related to Upper Paleolithic Siberians and contributed to both Europeans and Near Easterners; and Early European Farmers (EEF), who were mainly of Near Eastern origin but also harbored WHG-related ancestry. We model these populations' deep relationships and show that EEF had $\sim$44% ancestry from a "Basal Eurasian" lineage that split prior to the diversification of all other non-African lineages.
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            A revised timescale for human evolution based on ancient mitochondrial genomes.

            Recent analyses of de novo DNA mutations in modern humans have suggested a nuclear substitution rate that is approximately half that of previous estimates based on fossil calibration. This result has led to suggestions that major events in human evolution occurred far earlier than previously thought. Here, we use mitochondrial genome sequences from ten securely dated ancient modern humans spanning 40,000 years as calibration points for the mitochondrial clock, thus yielding a direct estimate of the mitochondrial substitution rate. Our clock yields mitochondrial divergence times that are in agreement with earlier estimates based on calibration points derived from either fossils or archaeological material. In particular, our results imply a separation of non-Africans from the most closely related sub-Saharan African mitochondrial DNAs (haplogroup L3) that occurred less than 62-95 kya. Though single loci like mitochondrial DNA (mtDNA) can only provide biased estimates of population divergence times, they can provide valid upper bounds. Our results exclude most of the older dates for African and non-African population divergences recently suggested by de novo mutation rate estimates in the nuclear genome. Copyright © 2013 Elsevier Ltd. All rights reserved.
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              Crop manuring and intensive land management by Europe's first farmers.

              The spread of farming from western Asia to Europe had profound long-term social and ecological impacts, but identification of the specific nature of Neolithic land management practices and the dietary contribution of early crops has been problematic. Here, we present previously undescribed stable isotope determinations of charred cereals and pulses from 13 Neolithic sites across Europe (dating ca. 5900-2400 cal B.C.), which show that early farmers used livestock manure and water management to enhance crop yields. Intensive manuring inextricably linked plant cultivation and animal herding and contributed to the remarkable resilience of these combined practices across diverse climatic zones. Critically, our findings suggest that commonly applied paleodietary interpretations of human and herbivore δ(15)N values have systematically underestimated the contribution of crop-derived protein to early farmer diets.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                20 January 2016
                2016
                : 11
                : 1
                : e0146176
                Affiliations
                [1 ]Danube Private University, Krems, Austria
                [2 ]Institute for Prehistory and Archaeological Science and Hightech Research Center, Basel University, Basel, Switzerland
                [3 ]State Office for Heritage Management and Archaeology Saxony-Anhalt and State Museum of Prehistory, Halle, Germany
                [4 ]German Mummy Project, Reiss-Engelhorn-Museen, Mannheim, Germany
                [5 ]Department of Prehistory and Archaeology, Faculty of Philosophy and Letters, Autonomous University of Madrid, Madrid, Spain
                [6 ]Curt Engelhorn Centre Archaeometry gGmbH, Mannheim, Germany
                [7 ]Laboratory of Archaeogenetics, Institute of Archaeology, Research Centre for the Humanities, Hungarian Academy of Sciences, Budapest, Hungary
                [8 ]Institute of Anthropology, Mainz University, Mainz, Germany
                [9 ]Arcadia-General Foundation of Valladolid University, Valladolid, Spain
                [10 ]Department of Applied and Analytical Paleontology, Mainz University, Mainz, Germany
                [11 ]Department of Prehistory, University of the Basque Government, Vitoria, Spain
                [12 ]Laboratoire TRACES UMR5608, Université de Toulouse, Toulouse, France
                [13 ]Private Technical Archaeologist, Zaragoza, Spain
                [14 ]Department of Prehistory and Archaeology, Valladolid University, Valladolid, Spain
                University of Otago, NEW ZEALAND
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: KWA MARG RGP. Performed the experiments: KWA CK PH CR DN. Analyzed the data: KWA ASN CR PH CK SZ MARG RGP HAM. Contributed reagents/materials/analysis tools: KWA MARG RGP CTR IGML HAM. Wrote the paper: KWA CK MARG RGP PH ASN CR SZ. Participated in the excavation: MARG CTR IGML. Provided ancient samples, contextual information, and access to unpublished archaeological data: MARG RGP CTR IGML HAM. Provided input on the manuscript: CTR IGML HAM.

                Article
                PONE-D-15-27139
                10.1371/journal.pone.0146176
                4720281
                26789731
                468a3975-e6a2-4021-a177-a65d3681f0a5
                © 2016 Alt et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                : 26 June 2015
                : 13 December 2015
                Page count
                Figures: 7, Tables: 1, Pages: 32
                Funding
                The archaeological research was supported and financed by the Junta de Castilla y León (Spain), the anthropological analyses by the German Research Foundation Al 287/14-1, the Johannes Gutenberg University of Mainz, Germany (no grant number), and the Danube Private University, Krems, Austria (no grant number). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Custom metadata
                All relevant data are within the paper and its Supporting Information files. The HVS-I mtDNA consensus sequences were uploaded to GenBank ( www.ncbi.nlm.nih.gov/genbank) under the accession numbers KT868907-KT868932.

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