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Includes supplementary materials for the online appendix. Human populations underwent range contractions during the Last Glacial Maximum (LGM) which had lasting and dramatic effects on their genetic variation. The genetic ancestry of individuals associated with the post-LGM Magdalenian technocomplex has been interpreted as being derived from groups associated with the pre-LGM Aurignacian. However, both these ancestries differ from that of central European individuals associated with the chronologically intermediate Gravettian. Thus, the genomic transition from pre- to post-LGM remains unclear also in western Europe, where we lack genomic data associated with the intermediate Solutrean, which spans the height of the LGM. Here we present genome-wide data from sites in Andalusia in southern Spain, including from a Solutrean-associated individual from Cueva del Malalmuerzo, directly dated to ~23,000 cal yr BP. The Malalmuerzo individual carried genetic ancestry that directly connects earlier Aurignacian-associated individuals with post-LGM Magdalenian-associated ancestry in western Europe. This scenario differs from Italy, where individuals associated with the transition from pre- and post-LGM carry different genetic ancestries. This suggests different dynamics in the proposed southern refugia of Ice Age Europe and posits Iberia as a potential refugium for western European pre-LGM ancestry. More, individuals from Cueva Ardales, which were thought to be of Palaeolithic origin, date younger than expected and, together with individuals from the Andalusian sites Caserones and Aguilillas, fall within the genetic variation of the Neolithic, Chalcolithic and Bronze Age individuals from southern Iberia. We thank all members of the Archaeogenetics Department of the Max Planck Institute for Evolutionary Anthropology, especially the Population Genetics group, and we thank Paleogenomics Lab members from Instituto de Biología Evolutiva for the data discussion. We especially thank M. Hajdinjak and I. Olalde for valuable comments and suggestions. We thank G. Brandt, A. Wissgott, S. Clayton and K. Prüfer for sequencing and handling the raw data. We thank all the members of the general research project in Cueva de Ardales and Sima de las Palomas de Teba. We thank all colleagues, archaeologists, geologists and experts in different scientific disciplines, as well as students from the Universities of Cádiz, Cologne and Tübingen, who have participated in the field and laboratory works. We are grateful for the great collaboration of the Town Councils of Ardales and Teba, within the framework of collaboration agreements with the University of Cadiz and the Neanderthal Museum. This work was supported by the Max Planck Society (V.V.-M., A.B.R., J.K. and W.H.), Margarita Salas 2022 funded by Unión Europea-Next Generation EU (V.V.-M.) and the European Research Council under the European Union’s Horizon 2020 research and innovation programme (grant agreement no. 771234-PALEoRIDER to W.H. and no. 803147-RESOLUTION to S.T.). The information provided on recent dating and archaeological records from Cueva de Ardales is part of the General Research Project, authorized by the Consejería de Cultura y Patrimonio Histórico de la Junta de Andalucía: entitled ‘Prehistoric societies (from the Middle Palaeolithic to the Final Neolithic) in the Cueva de Ardales and Sima de las Palomas de Teba (Málaga, Spain)’. A geoarchaeological, chronological and environmental study was conducted under the direction of J.R.-M. and G.-C.W. between 2015 and 2021. Excavations and field and laboratory studies have been carried out in Cueva de Ardales within the framework of the projects entitled ‘Analysis of prehistoric societies from the Middle Palaeolithic to the Late Neolithic on the two shores of the Strait of Gibraltar’. Relaciones y contactos—HAR2017-8734P (Ministerio de Economía, Industria y Competitividad-Agencia Estatal de Investigación, cofinanced by FEDER funds) for which J.R.-M. and Salvador Domínguez-Bella are the responsible researchers. Collaborative Research Centre—CRC 806 Our Way to Europe, funded by the Deutsche Forschungsgemeinschaft (DFG-German Research Foundation). Project no. 57444011, under the responsibility of G.-C.W.

Acknowledgements: The authors thank G. Marciani and O. Jöris for comments on archaeology; C. Jeong, M. Spyrou and K. Prüfer for comments on genetics; M. O’Reilly for graphical support for Fig. 5 and Extended Data Fig. 9; the entire IT and laboratory teams at the Department of Archaeogenetics of MPI-SHH for technical assistance; M. Meyer and S. Nagel for support with single-stranded library preparation; K. Post, P. van Es, J. Glimmerveen, M. Medendorp, M. Sier, S. Dikstra, M. Dikstra, R. van Eerden, D. Duineveld and A. Hoekman for providing access to human specimens from the North Sea (The Netherlands); M. D. Garralda and A. Estalrrich for providing access to human specimens from La Riera (Spain); J. Górski and M. Zając for providing access to human specimens from Maszycka cave; C. Di Patti for providing access to human specimens from San Teodoro 2 (Italy); P. Blaževičius for providing access to the Donkalnis human remains and the new radiocarbon dates; the Italian Ministry of Culture and Soprintendenza Archeologia Belle Arti e Paesaggio for the Provinces of Verona, Rovigo, and Vicenza for granting access to the human remains of Tagliente 2; F. Fontana, who carries out investigations of the Riparo Tagliente site (Italy); the Friuli Venezia Giulia Superintendency for providing access to the human tooth Pradis 1; and the Soprintendenza Archeologia Belle Arti e Paesaggio for the Provinces of Barletta-Andria-Trani and Foggia for providing access to the Paglicci human remains. This project has received funding by the European Research Council under the European Union’s Horizon 2020 research and innovation programme under grant agreements no. 803147-RESOLUTION (to S.T.), no. 771234-PALEoRIDER (to W.H.), no. 864358 (to K.M.), no. 724703 and no. 101019659 (to K.H.). K.H. is also supported by the Deutsche Forschungsgemeinschaft (DFG FOR 2237). E.A. has received funding from the Van de Kamp fonds. PACEA co-authors of this research benefited from the scientific framework of the University of Bordeaux’s IdEx Investments for the Future programme/GPR Human Past. A.G.-O. is supported by a Ramón y Cajal fellowship (RYC-2017-22558). L. Sineo, M.L. and D.C. have received funding from the Italian Ministry of University and Research (MUR) PRIN 2017 grants 20177PJ9XF and 20174BTC4R_002. H. Rougier received support from the College of Social and Behavioral Sciences of CSUN and the CSUN Competition for RSCA Awards. C.L.S. and T. Saupe received support from the European Union through the European Regional Development Fund (project no. 2014-2020.4.01.16-0030) and C.L.S. received support from the Estonian Research Council grant PUT (PRG243). S. Shnaider received support from the Russian Science Foundation (no. 19-78-10053)., 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.

The recent discovery of an exceptionally rich grave at La Almoloya in south-eastern Spain illuminates the political context of Early Bronze Age El Argar society. The quantity, variety and opulence of the grave goods emphasise the technological, economic and social dimensions of this unique culture. The assemblage includes politically and ideologically emblematic objects, among which a silver diadem stands out. Of equally exceptional character is the building under which the grave was found—possibly one of the first Bronze Age palaces identified in Western Europe. The architecture and artefacts from La Almoloya provide new insight into emblematic individuals and the exercise of power in societies of marked economic asymmetry. Introduction Argaric diadems as embodied emblems La Almoloya grave 38 Spaces of power Conclusions

The characterization of Neandertals’ diets has mostly relied on nitrogen isotope analyses of bone and tooth collagen. However, few nitrogen isotope data have been recovered from bones or teeth from Iberia due to poor collagen preservation at Paleolithic sites in the region. Zinc isotopes have been shown to be a reliable method for reconstructing trophic levels in the absence of organic matter preservation. Here, we present the results of zinc (Zn), strontium (Sr), carbon (C), and oxygen (O) isotope and trace element ratio analysis measured in dental enamel on a Pleistocene food web in Gabasa, Spain, to characterize the diet and ecology of a Middle Paleolithic Neandertal individual. Based on the extremely low δZn value observed in the Neandertal’s tooth enamel, our results support the interpretation of Neandertals as carnivores as already suggested by δN isotope values of specimens from other regions. Further work could help identify if such isotopic peculiarities (lowest δZn and highest δN of the food web) are due to a metabolic and/or dietary specificity of the Neandertals., This study was funded by Deutsche Forschungsgemeinschaft Project PALEODIET (378496604), European Research Council (ERC) Project ARCHEIS (Grant 803676), ERC Project VERTEBRATE HERBIVORY (Grant 681450), and the Max Planck Society. G.M.S. received funding from the European Union’s Horizon Europe Research and Innovation Programme under Marie Skłodowska–Curie Grant Agreement 101027850. We thank Maria Alonso Lescún from the Museum of Huesca for her help during the collection sampling, Adeline Le Cabec for her help in understanding of tooth development, Sven Steinbrenner for technical help, Mary Kita for her help in the improvement of the figures, Jeremy McCormack for helpful discussions, and Hubert Vonhof for access to the stable isotope lab at MPIC Mainz. D.C.S.G. acknowledges funding by the Generalitat Valenciana (CIDEGENT/2019/061), and L.M. and P.U. from the Ministerio de Ciencia e Innovación (MICINN) (PID2020-116598GB-I00 GAPS AND DATES. DINAMICAS CULTURALES EN LA PREHISTORIA DE LA CUENCA DEL EBRO).

Uniparentally-inherited markers on mitochondrial DNA (mtDNA) and the non-recombining regions of the Y chromosome (NRY), have been used for the past 30 years to investigate the history of humans from a maternal and paternal perspective. Researchers have preferred mtDNA due to its abundance in the cells, and comparatively high substitution rate. Conversely, the NRY is less susceptible to back mutations and saturation, and is potentially more informative than mtDNA owing to its longer sequence length. However, due to comparatively poor NRY coverage via shotgun sequencing, and the relatively low and biased representation of Y-chromosome variants on capture assays such as the 1240 k, ancient DNA studies often fail to utilize the unique perspective that the NRY can yield. Here we introduce a new DNA enrichment assay, coined YMCA (Y-mappable capture assay), that targets the "mappable" regions of the NRY. We show that compared to low-coverage shotgun sequencing and 1240 k capture, YMCA significantly improves the mean coverage and number of sites covered on the NRY, increasing the number of Y-haplogroup informative SNPs, and allowing for the identification of previously undiscovered variants. To illustrate the power of YMCA, we show that the analysis of ancient Y-chromosome lineages can help to resolve Y-chromosomal haplogroups. As a case study, we focus on H2, a haplogroup associated with a critical event in European human history: the Neolithic transition. By disentangling the evolutionary history of this haplogroup, we further elucidate the two separate paths by which early farmers expanded from Anatolia and the Near East to western Europe., Open Access funding enabled and organized by Projekt DEAL. This study was funded by the Max Planck Society, the French (ANR) and German (DFG) Research Foundations under the INTERACT project (ANR-17-FRAL-0010, DFG-HA-5407/4-1, 2018-2021) to M.R. and W.H., the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program under Grant agreement no. 771234-PALEoRIDER to W.H., the award Praemium Academiae of the Czech Academy of Sciences to M.E. and the project RVO 67985912 of the Institute of Archaeology of the Czech Academy of Sciences, Prague to M.S.

We present a very intriguing set of finds coming from Tragaluz Norte, one of the most inaccessible rock shelters in the Vero canyon, which runs north to south through the Sierra de Guara, in the Huesca province (north-east Spain). There, a skeleton from a 9-years-old child with his toy wooden bow as well as other elements (pieces of wood, fabric and pottery, and a stone wall) were found. The whole ensemble belongs to a single moment in the second half of the 4th century CE. We hypothesize that this child was part of a group of rural laborers who died accidentally and was buried there. As an explanation for their presence in such an inaccessible place we think that they could be gathering honey from man-made cane and mud honeycombs -locally called arnas-, a task documented in the area until recent times.

Description, Ancient human DNA from southern Iberia reveals social and genomic changes during the emergence of the Bronze Age in Europe., The emerging Bronze Age (BA) of southeastern Iberia saw marked social changes. Late Copper Age (CA) settlements were abandoned in favor of hilltop sites, and collective graves were largely replaced by single or double burials with often distinctive grave goods indirectly reflecting a hierarchical social organization, as exemplified by the BA El Argar group. We explored this transition from a genomic viewpoint by tripling the amount of data available for this period. Concomitant with the rise of El Argar starting ~2200 cal BCE, we observe a complete turnover of Y-chromosome lineages along with the arrival of steppe-related ancestry. This pattern is consistent with a founder effect in male lineages, supported by our finding that males shared more relatives at sites than females. However, simple two-source models do not find support in some El Argar groups, suggesting additional genetic contributions from the Mediterranean that could predate the BA.

Hepatitis B virus (HBV) has been infecting humans for millennia and remains a global health problem, but its past diversity and dispersal routes are largely unknown. We generated HBV genomic data from 137 Eurasians and Native Americans dated between ~10,500 and ~400 years ago. We date the most recent common ancestor of all HBV lineages to between ~20,000 and 12,000 years ago, with the virus present in European and South American hunter-gatherers during the early Holocene. After the European Neolithic transition, Mesolithic HBV strains were replaced by a lineage likely disseminated by early farmers that prevailed throughout western Eurasia for ~4000 years, declining around the end of the 2nd millennium BCE. The only remnant of this prehistoric HBV diversity is the rare genotype G, which appears to have reemerged during the HIV pandemic., The research was funded by the Max Planck Society, the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (771234–PALEoRIDER, to W.H.; 805268–CoDisEASe to K. Bos; 834616–ARCHCAUCASUS to S.H.), the Slovak Academy of Sciences and the European Union’s Seventh Framework Programme and Marie Curie Actions under the Programme SASPRO (1340/03/03 to P.C.R.), the ERA.NET RUS Plus–S&T programm of the European Union’s Seventh Framework Programme (277–BIOARCCAUCASUS to S.Re. and S.H.), the Werner Siemens Stiftung (“Paleobiochemistry”, to CW), the Award Praemium Academiae of the Czech Academy of Sciences (to M.E.), the Institute of Archaeology of the Czech Academy of Sciences (RVO 67985912, to M.Dobe.), the Russian Foundation for Basic Research (19-09-00354a, to M.K.K. and V.V.K.; 19-78-10053 to SSh), the German Research Foundation (DFG-HA-5407/4-1–INTERACT to W.H. and RE2688/2 to S.Re.), the French National Research Agency (ANR-17-FRAL-0010–INTERACT, to M.F.D., M.Ri., S.Ro., S.Sai., D.Bi., and P.Le.), the Wenner-Gren Dissertation Fieldwork Grant (9558 to S.Sab.), and the Ministry of Education and Science of the Republic of Kazakhstan (AP08856654 to L.B.D., L.M., and E.Kh. and AP08857177 to A.Z.B.).

Southern Italy is a key region for understanding the agricultural transition in the Mediterranean due to its central position. We present a genomic transect for 19 prehistoric Sicilians that covers the Early Mesolithic to Early Neolithic period. We find that the Early Mesolithic hunter-gatherers (HGs) are a highly drifted sister lineage to Early Holocene western European HGs, whereas a quarter of the Late Mesolithic HGs ancestry is related to HGs from eastern Europe and the Near East. This indicates substantial gene flow from (south-)eastern Europe between the Early and Late Mesolithic. The Early Neolithic farmers are genetically most similar to those from the Balkan and Greece, and carry only a maximum of ∼7% ancestry from Sicilian Mesolithic HGs. Ancestry changes match changes in dietary profile and material culture, except for two individuals who may provide tentative initial evidence that HGs adopted elements of farming in Sicily.One-sentence summaryGenome-wide and isotopic data from prehistoric Sicilians reveal a pre-farming connection to (south-) eastern Europe, and tentative initial evidence that hunter-gatherers adopted some Neolithic aspects prior to near-total replacement by early farmers.

There is an increasing abundance in the archaeological record in Iberia for the Late Neolithic and the beginning of the Chalcolithic periods, mostly regarding burials. The higher pre-Pyrenean areas began to be settled more frequently, but the poor weather conditions have led researchers to suggest that human presence mostly took the form of sporadic visits. This argument has provoked substantial controversy given the increase not only in the archaeological artefacts recorded but also in the number of burial sites in less accessible places. To shed more light on the knowledge of these Chalcolithic mountain groups, we have carried out strontium isotope analysis of human enamel of individuals from the funerary cave Cueva de los Cristales (Sarsa de Surta, Huesca, Espana), located 1,300 m above sea level (m a.s.l.). Our results point to a pre-Pyrenean origin of the Chalcolithic groups, despite differences found among the individual values, which may be related to different locations in the pre-Pyrenean area. The added value of the study resides in the large amount of data points (up to 40) of bioavailable strontium values which will be useful for future mobility studies performed in the Pyrenean territory.

Carbon (δ13C) and nitrogen (δ15N) stable isotope analyses from bone collagen provide information about the dietary protein input, while strontium isotopes (87Sr/86Sr) from tooth enamel give us data about provenance and potential territorial mobility of past populations. To date, isotopic results on the prehistory of the Western Pyrenees are scarce. In this article, we report human and faunal values of the mentioned isotopes from the Early-Middle Neolithic site of Fuente Hoz (Anuntzeta) and the Late Neolithic/Early Chalcolithic site of Kurtzebide (Letona, Zigoitia). The main objectives of this work are to analyse the dietary and territorial mobility patterns of these populations. Furthermore, as an additional aim, we will try to discuss social ranking based on the isotope data and existing literature on this topic in the region of study. Our results show that, based on the bioavailable Sr values, both purported local and non-local humans were buried together at the sites. Additionally, they suggest similar resource consumption based on C3 terrestrial resources (i.e. ovicaprids, bovids, and suids) as the main part of the protein input. Overall, this study sheds light on how individuals from different backgrounds were still buried together and shared the same “dietary lifestyle” at a time in the Prehistory of Iberia when social complexities started to appear.

The use of isotopic analysis in human and animal remains from the Holocene has proved to be a very useful tool to explore the exploitation and adaptation of past populations to different environments. In this study we present isotopic analysis results of carbon, nitrogen and strontium from the Late Neolithic-Chalcolithic site of San Juan cave (Loarre, Spain). We analysed 33 humans, divided in adult and subadult groups, and 16 animals recovered from the same archaeological context. Stable isotope analysis of carbon and nitrogen has allowed to distinguish an homogeneous subsistence pattern during the Late Neolithic-Chalcolithic transition. The use of strontium isotopes (87Sr/86Sr) in human dental enamel suggests 19% (4 out of 21) are non-local individuals, based on comparison with the local bioavailable 87Sr/86Sr range calculated using microfauna teeth from the archaeological context, modern plants and snails. This new study gives information about Late Neolithic communities located in the north-east of the Iberian Peninsula, and it allows inference of the socio-economic structure, territorial mobility and individual provenance of humans. Fil: Villalba Mouco, Vanessa. Universidad de Zaragoza; España Fil: Sauqué Latas, Víctor. Universidad de Zaragoza; España. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Antropología de Córdoba. Universidad Nacional de Córdoba. Facultad de Filosofía y Humanidades. Instituto de Antropología de Córdoba; Argentina Fil: Sarasketa Gartzia, Izaskun. Universidad del País Vasco; España Fil: Pastor, M. Victoria. Universidad de Zaragoza; España Fil: le Roux, Petrus J.. Cape Town Universtity; Sudáfrica Fil: Vicente, Diana. Universidad de Zaragoza; España Fil: Utrilla, Pilar. Universidad de Zaragoza; España Fil: Salazar García, Domingo C.. Cape Town Universtity; Sudáfrica. Universidad del País Vasco; España

The Late Neolithic and Chalcolithic periods are poorly understood in northeastern Iberia. Most of the information comes from the sepulchral structures rather than habitat settlements. The high number of individuals usually recovered from this types of collective burial spaces, together with the low number of direct radiocarbon dates available on them, forces us to be cautious and consider all the studied assemblages as belonging to the so-called Late Neolithic-Chalcolithic time period. To evaluate human dietary patterns of the Late Neolithic-Chalcolithic populations from the northeast of Iberia, stable carbon and nitrogen isotope analysis was carried out on 78 humans and 32 faunal bones from Cova de la Guineu (Font-rubi, Barcelona) and Cueva de Abauntz (Arraitz, Navarra), both of them sepulchral sites. Results show a common dietary pattern in both sites, indicating an homogeneous protein diet based on C3 terrestrial resources and no isotopic evidence of the consumption of C4 plants. Only one individual from Cueva de Abauntz, who directly dates to the first moments of the use of the cave as a burial place, suggests a different protein intake. The inter-population analysis shows a significant difference between both human and faunal δ13C values, suggesting an environmental influence on the isotope values depending on the geographic location. This effect should not be discarded and always assessed with baseline isotopic values in future studies at each area of Iberia and for different chronological moments.

The Iberian Peninsula in southwestern Europe represents an important test case for the study of human population movements during prehistoric periods. During the Last Glacial Maximum (LGM), the peninsula formed a periglacial refugium [1] for hunter-gatherers (HGs) and thus served as a potential source for the re-peopling of northern latitudes [2]. The post-LGM genetic signature was previously described as a cline from Western HG (WHG) to Eastern HG (EHG), further shaped by later Holocene expansions from the Near East and the North Pontic steppes [3, 4, 5, 6, 7, 8, 9]. Western and central Europe were dominated by ancestry associated with the ∼14,000-year-old individual from Villabruna, Italy, which had largely replaced earlier genetic ancestry, represented by 19,000–15,000-year-old individuals associated with the Magdalenian culture [2]. However, little is known about the genetic diversity in southern European refugia, the presence of distinct genetic clusters, and correspondence with geography. Here, we report new genome-wide data from 11 HGs and Neolithic individuals that highlight the late survival of Paleolithic ancestry in Iberia, reported previously in Magdalenian-associated individuals. We show that all Iberian HGs, including the oldest, a ∼19,000-year-old individual from El Mirón in Spain, carry dual ancestry from both Villabruna and the Magdalenian-related individuals. Thus, our results suggest an early connection between two potential refugia, resulting in a genetic ancestry that survived in later Iberian HGs. Our new genomic data from Iberian Early and Middle Neolithic individuals show that the dual Iberian HG genomic legacy pertains in the peninsula, suggesting that expanding farmers mixed with local HGs., The genetic research was funded by the Max Planck Society and the European Research Council ERC-CoG 771234 PALEoRIDER (W.H.). V.V.-M. was funded by a predoctoral scholarship of the Gobierno de Aragon and the Fondo Social Europeo (BOA20150701025) and a 3-month research stay grant (CH 76/16) by Programa CAI-Ibercaja de Estancias de Investigacio´ n. V.V.-M. and P.U. are members of the Spanish project HAR2014-59042-P (Transiciones clima´ ticas y adaptaciones sociales en la prehistoria de la Cuenca del Ebro), and of the regional government of Aragon PPVE research group (H-07: Primeros Pobladores del Valle del Ebro). The Goyet project was funded by the Wenner-Gren Foundation (7837 to H.R.), the College of Social and Behavioral Sciences of CSUN, and the CSUN Competition for Research, Scholarship, and Creative Activity Awards.

We assembled genome-wide data from 271 ancient Iberians, of whom 176 are from the largely unsampled period after 2000 BCE, thereby providing a high-resolution time transect of the Iberian Peninsula. We document high genetic substructure between northwestern and southeastern hunter-gatherers before the spread of farming. We reveal sporadic contacts between Iberia and North Africa by ~2500 BCE and, by ~2000 BCE, the replacement of 40% of Iberia's ancestry and nearly 100% of its Y-chromosomes by people with Steppe ancestry. We show that, in the Iron Age, Steppe ancestry had spread not only into Indo-European-speaking regions but also into non-Indo-European-speaking ones, and we reveal that present-day Basques are best described as a typical Iron Age population without the admixture events that later affected the rest of Iberia. Additionally, we document how, beginning at least in the Roman period, the ancestry of the peninsula was transformed by gene flow from North Africa and the eastern Mediterranean., J.M.F., F.J.L.-C., J.I.M., F.X.O., J.D., and M.S.B. were supported by HAR2017-86509-P, HAR2017-87695-P, and SGR2017-11 from the Generalitat de Catalunya, AGAUR agency. C.L.-F. was supported by Obra Social La Caixa and by FEDER-MINECO (BFU2015- 64699-P). L.B.d.L.E. was supported by REDISCO-HAR2017-88035-P (Plan Nacional I+D+I, MINECO). C.L., P.R., and C.Bl. were supported by MINECO (HAR2016-77600-P). A.Esp., J.V.-V., G.D., and D.C.S.-G. were supported by MINECO (HAR2009-10105 and HAR2013-43851-P). D.J.K. and B.J.C. were supported by NSF BCS-1460367. K.T.L., A.W., and J.M. were supported by NSF BCS-1153568. J.F.-E. and J.A.M.-A. were supported by IT622-13 Gobierno Vasco, Diputación Foral de Álava, and Diputación Foral de Gipuzkoa. We acknowledge support from the Portuguese Foundation for Science and Technology (PTDC/EPH-ARQ/4164/2014) and the FEDER-COMPETE 2020 project 016899. P.S. was supported by the FCT Investigator Program (IF/01641/2013), FCT IP, and ERDF (COMPETE2020 – POCI). M.Si. and K.D. were supported by a Leverhulme Trust Doctoral Scholarship awarded to M.B.R. and M.P. D.R. was supported by an Allen Discovery Center grant from the Paul Allen Foundation, NIH grant GM100233, and the Howard Hughes Medical Institute. V.V.-M. and W.H. were supported by the Max Planck Society.

Objectives The Early Neolithic involved an important social and economic shift that can be tested not only with the material culture, but also through biomolecular approaches. The Iberian Peninsula presents few Early Neolithic sites where fauna and humans can be analyzed together from an isotopic perspective. Here we present an isotopic study on the site of Cueva de Chaves as an example for understanding the dietary and economical changes that took place during Early Neolithic in Iberia. Material and methods Here we apply carbon and nitrogen stable isotope analysis to bone collagen from 4 humans and 64 faunal samples from 14 different species. The large dataset belongs to the same unique chrono-cultural context secured by 20 radiocarbon dates. Three direct new radiocarbon dates were carried out on the human remains analyzed. Results Faunal isotope values show no significant differences between wild and domestic herbivores, although the latter have more homogeneous values. Domestic pigs, potentially considered omnivorous, also show signatures of a herbivore diet. Human isotopic results show a diet mainly based on terrestrial C3 resources and possibly high meat consumption. The only individual found buried with a special funerary treatment presents a slightly different protein intake, when taking into account the long contemporaneous baseline analyzed. Discussion Similar values between wild and domestic species could be the result of common feeding resources and/or grazing on the same parts of the landscape. The herbivore diet seen amongst domestic pigs rules out feeding on household leftovers. High meat consumption by humans would support the hypothesis of the existence of a specialized animal husbandry management community in which agriculture was not intensively developed. Our results suggest that the development of agricultural practices and animal husbandry were not necessarily associated together in the early stages of the Western Mediterranean Neolithic.

However, we are afraid that there was a misunderstanding in the interpretation of our data. Ben-Dor and Barkai (2) wrote that "One explanation presented in the [our] paper was that Neandertals may have consumed body parts and animals that were relatively poor in zinc, like the liver or deer." This sentence is not reflecting our conclusions: We discussed Zn isotope ratios (i.e., 66Zn/64Zn ratio expressed as a δ66Zn value) and not Zn concentrations. To explain low δ66Zn values of the Neandertal tooth, we propose that Neandertals ate food items depleted in heavy Zn isotopes (and therefore enriched in light Zn isotopes), but those foods can have variable Zn contents, independent of the Zn isotope composition as such.