Genomic ancestry and social dynamics of the last hunter-gatherers of Atlantic France.

Since the early Holocene, western and central Europe was inhabited by a genetically distinct group of Western Hunter-Gatherers (WHGs). This group was eventually replaced and assimilated by the incoming Neolithic farmers. The western Atlantic façade was home to some of the last Mesolithic sites of mainland Europe, represented by the iconic open-air sites at Hoedic and Téviec in southern Brittany, France. These sites are known for the unusually well-preserved and rich burials. Genomic studies of Mesolithic European hunter-gatherers have been limited to single or a few individuals per site and our understanding of the social dynamics of the last Mesolithic hunter-gatherers of Europe and their interactions with incoming farmers is limited. We sequenced and analyzed the complete genomes of 10 individuals from the Late Mesolithic sites of Hoedic, Téviec, and Champigny, in France, four of which sequenced to between 23- and 8-times genome coverage. The analysis of genomic, chronological and dietary data revealed that the Late Mesolithic populations in Brittany maintained distinct social units within a network of exchanging mates. This resulted in low intra-group biological relatedness that prevented consanguineous mating, despite the small population size of the Late Mesolithic groups. We found no genetic ancestry from Neolithic farmers in the analyzed hunter-gatherers, even though some of them may have coexisted with the first farming groups in neighboring regions. Hence, contrary to previous conclusions based on stable isotope data from the same sites, the Late Mesolithic forager community was limited in mate-exchange to neighboring hunter-gatherer groups, to the exclusion of Neolithic farmers.

The Ornaments of the Arma Veirana Early Mesolithic Infant Burial.

Personal ornaments are widely viewed as indicators of social identity and personhood. Ornaments are ubiquitous from the Late Pleistocene to the Holocene, but they are most often found as isolated objects within archaeological assemblages without direct evidence on how they were displayed. This article presents a detailed record of the ornaments found in direct association with an Early Mesolithic buried female infant discovered in 2017 at the site of Arma Veirana (Liguria, Italy). It uses microscopic, 3D, and positional analyses of the ornaments as well as a preliminary perforation experiment to document how they were perforated, used, and what led to their deposit as part of the infant's grave goods. This study provides important information on the use of beads in the Early Mesolithic, in general, as well as the relationship between beads and young subadults, in particular. The results of the study suggest that the beads were worn by members of the infant's community for a considerable period before they were sewn onto a sling, possibly used to keep the infant close to the parents while allowing their mobility, as seen in some modern forager groups. The baby was then likely buried in this sling to avoid reusing the beads that had failed to protect her or simply to create a lasting connection between the deceased infant and her community. Supplementary Information: The online version contains supplementary material available at 10.1007/s10816-022-09573-7.

Mesolithic hearth-pits and formation processes: a geoarchaeological investigation of sediments from El Arenal de la Virgen site (SE Iberia).

Hearth-pits are some of the most common archaeological features documented in open-air Mesolithic sites, especially in coversand areas of NW Europe. However, very few geoarchaeological studies have addressed their formation, function and relationship with occupation surfaces. This work introduces new interdisciplinary investigations on the sediments of the Mesolithic open-air site of El Arenal de la Virgen (SE Iberia). A selection of five hearth-pits from two different occupation phases (Phase 1: 9.3-9.1 cal ka BP and Phase 2: 8.6-8.3 cal ka BP) has been analysed using stratigraphy, texture, soil chemistry, micromorphology, petrography and OSL and TL analyses. Combustion traits of the carbonate rock assemblages preserved in the sediments of the hearth-pits have also been investigated and compared to reference and experimental data from local geogenic materials. Our results allowed us to discuss the anthropogenic origin and taphonomy of the hearth-pits studied and approach their function. The structures from Phase 1 are interpreted as a possible oven and a dumping feature linked to single/occasional use events. In contrast, for hearth-pits from Phase 2, we propose they were related to combustion and dwelling areas subject to recurrent occupation episodes and disturbance. Finally, our sedimentary and soil data revealed existing favourable paleoenvironmental conditions during the Mesolithic occupation of the site characterized by increased moisture, temperature and vegetation cover, in contrast to the Pleistocene and Middle Holocene periods pre- and post-dating the human settlement. This work highlights the potential of integrating geoarchaeological and contextual evidence to clarify the factors involved in the formation of hearth-pits and infer intra-site occupation patterns. Supplementary information: The online version contains supplementary material available at 10.1007/s12520-023-01794-5.

Carbon, nitrogen and sulphur isotope data of archaeological fish and mammal bone collagen from Lithuania.

Until relatively recently, stable sulphur isotope analysis of bone collagen was seldom undertaken in bioarchaeological research. With increasing frequency, its application has proven useful in reconstructing palaeodiets and palaeoecologies, as well as identifying potential migration and mobility patterns. Here, sulphur (δ34S) isotope analysis, together with carbon (δ13C) and nitrogen (δ15N), was performed on six fish and 34 mammal bone collagen samples from 14 prehistoric sites in Lithuania dating from the Late Mesolithic (ca. 7000-5000 cal BC) to the Late Bronze Age (ca. 1100-500 cal BC). We present the first δ34S data from Lithuania, including coupled δ13C and δ15N data, offering a crucial dataset for future research to explore spatial and temporal variability in the region and beyond.

Hunter-gatherer admixture facilitated natural selection in Neolithic European farmers.

Ancient DNA has revealed multiple episodes of admixture in human prehistory during geographic expansions associated with cultural innovations. One important example is the expansion of Neolithic agricultural groups out of the Near East into Europe and their consequent admixture with Mesolithic hunter-gatherers.1,2,3,4 Ancient genomes from this period provide an opportunity to study the role of admixture in providing new genetic variation for selection to act upon, and also to identify genomic regions that resisted hunter-gatherer introgression and may thus have contributed to agricultural adaptations. We used genome-wide DNA from 677 individuals spanning Mesolithic and Neolithic Europe to infer ancestry deviations in the genomes of admixed individuals and to test for natural selection after admixture by testing for deviations from a genome-wide null distribution. We find that the region around the pigmentation-associated gene SLC24A5 shows the greatest overrepresentation of Neolithic local ancestry in the genome (|Z| = 3.46). In contrast, we find the greatest overrepresentation of Mesolithic ancestry across the major histocompatibility complex (MHC; |Z| = 4.21), a major immunity locus, which also shows allele frequency deviations indicative of selection following admixture (p = 1 × 10-56). This could reflect negative frequency-dependent selection on MHC alleles common in Neolithic populations or that Mesolithic alleles were positively selected for and facilitated adaptation in Neolithic populations to pathogens or other environmental factors. Our study extends previous results that highlight immune function and pigmentation as targets of adaptation in more recent populations to selection processes in the Stone Age.

Stable isotope analyses of amino acids reveal the importance of aquatic resources to Mediterranean coastal hunter-gatherers.

Determining the degree to which humans relied on coastal resources in the past is key for understanding long-term social and economic development, as well as for assessing human health and anthropogenic impacts on the environment. Prehistoric hunter-gatherers are often assumed to have heavily exploited aquatic resources, especially those living in regions of high marine productivity. For the Mediterranean, this view has been challenged, partly by the application of stable isotope analysis of skeletal remains which has shown more varied coastal hunter-gatherer diets than in other regions, perhaps due to its lower productivity. By undertaking a more specific analysis of amino acids from bone collagen of 11 individuals from one of the oldest and best-known Mesolithic cemeteries in the Mediterranean, at El Collado, Valencia, we show that high levels of aquatic protein consumption were achieved. By measuring both carbon and nitrogen in amino acids, we conclude that some of the El Collado humans relied heavily on local lagoonal fish and possibly shellfish, rather than open marine species. By contrast to previous suggestions, this study demonstrates that the north-western coast of the Mediterranean basin could support maritime-oriented economies during the Early Holocene.

Wild cereal grain consumption among Early Holocene foragers of the Balkans predates the arrival of agriculture.

Forager focus on wild cereal plants has been documented in the core zone of domestication in southwestern Asia, while evidence for forager use of wild grass grains remains sporadic elsewhere. In this paper, we present starch grain and phytolith analyses of dental calculus from 60 Mesolithic and Early Neolithic individuals from five sites in the Danube Gorges of the central Balkans. This zone was inhabited by likely complex Holocene foragers for several millennia before the appearance of the first farmers ~6200 cal BC. We also analyzed forager ground stone tools (GSTs) for evidence of plant processing. Our results based on the study of dental calculus show that certain species of Poaceae (species of the genus Aegilops) were used since the Early Mesolithic, while GSTs exhibit traces of a developed grass grain processing technology. The adoption of domesticated plants in this region after ~6500 cal BC might have been eased by the existing familiarity with wild cereals.

Before humans invented agriculture and the first farmers appeared in southwestern Asia, other ancient foragers (also known as hunter-gatherers) in southeastern Europe had already developed a taste for consuming wild plants. There is evidence to suggest that these foragers were intensely gathering wild cereal grains before the arrival of agriculture. However, until now, the only place outside southwestern Asia this has been shown to have occurred is in Greece, and is dated around 20,000 years ago. In the past, researchers proposed that forager societies in the Balkans also consumed wild cereals before transitioning to agriculture. But this has been difficult to prove because plant foods are less likely to preserve than animal bones and teeth, making them harder to detect in prehistoric contexts. To overcome this, Cristiani et al. studied teeth from 60 individuals found in archaeological sites between Serbia and Romania, which are attributed to the Mesolithic and Early Neolithic periods. Food particles extracted from crusty deposits on the teeth (called the dental calculus) were found to contain structures typically found in plants. In addition, Cristiani et al. discovered similar plant food residues on ground stone tools which also contained traces of wear associated with the processing of wild cereals. These findings suggest that foragers in the central Balkans were already consuming certain species of wild cereal grains 11,500 years ago, before agriculture arrived in Europe. It is possible that sharing knowledge about plant resources may have helped introduce domesticated plant species in to this region as early as 6500 BC. This work challenges the deep-rooted idea that the diet of hunter-gatherers during the Palaeolithic and Mesolithic periods primarily consisted of animal proteins. In addition, it highlights the active role the eating habits of foragers might have played in introducing certain domesticated plant species that have become primary staples of our diet today.

Where Asia meets Europe - recent insights from ancient human genomics.

CONTEXT: The peopling of Europe by modern humans is a widely debated topic in the field of modern and ancient genomics. While several recent syntheses have focussed on this topic, little has been discussed about the genetic history of populations in the continent's surrounding regions. OBJECTIVE: We explore genetic transformations in three key areas that played an essential role in the formation of the European genetic landscape through time, focussing on the periods spanning from the Epipalaeolithic/Mesolithic and up until the Iron Age. METHODS: We review published ancient genomic studies and integrate the associated data to provide a quantification and visualisation of major trends in the population histories of the Near East, the western Eurasian Steppe and North East Europe. RESULTS: We describe cross-regional as well as localised prehistoric demographic shifts and discuss potential research directions while highlighting geo-temporal gaps in the data. CONCLUSION: In recent years, archaeogenetic studies have contributed to the understanding of human genetic diversity through time in regions located at the doorstep of Europe. Further studies focussing on these areas will allow for a better characterisation of genetic shifts and regionally-specific patterns of admixture across western Eurasia.

The use of lithic raw materials at the Early Mesolithic open-air site Feuersteinacker (Vogelsbergkreis, Germany).

The open-air site Feuersteinacker near Stumpertenrod has yielded one of the largest lithic assemblages in Central Germany. It repeatedly served as a workshop for the production of stone tools during an early phase of the Mesolithic. The range of lithic raw materials is extremely diverse, but until today, there is only a limited number of archaeological studies on the occurrence of lithic resources in the area. The following study presents the first in-depth investigation of the use of different rock types by Mesolithic hunter-fisher-gatherers at the site. Provenance analyses using petrographic methods permit raw materials to be assigned to a specific source and provide new insights into their formation. Furthermore, this study explores the way in which the materials were processed throughout the reduction sequence. A comparison of topographic parameters suggests that the location was situated on an important transit route during prehistoric times. The presented results contribute to a better understanding of mobility patterns and subsistence strategies of Early Mesolithic groups in Central Germany.

Heterogeneous Hunter-Gatherer and Steppe-Related Ancestries in Late Neolithic and Bell Beaker Genomes from Present-Day France.

The transition from the Late Neolithic to the Bronze Age has witnessed important population and societal changes in western Europe.1 These include massive genomic contributions of pastoralist herders originating from the Pontic-Caspian steppes2,3 into local populations, resulting from complex interactions between collapsing hunter-gatherers and expanding farmers of Anatolian ancestry.4-8 This transition is documented through extensive ancient genomic data from present-day Britain,9,10 Ireland,11,12 Iberia,13 Mediterranean islands,14,15 and Germany.8 It remains, however, largely overlooked in France, where most focus has been on the Middle Neolithic (n = 63),8,9,16 with the exception of one Late Neolithic genome sequenced at 0.05× coverage.16 This leaves the key transitional period covering ∼3,400-2,700 cal. years (calibrated years) BCE genetically unsampled and thus the exact time frame of hunter-gatherer persistence and arrival of steppe migrations unknown. To remediate this, we sequenced 24 ancient human genomes from France spanning ∼3,400-1,600 cal. years BCE. This reveals Late Neolithic populations that are genetically diverse and include individuals with dark skin, hair, and eyes. We detect heterogeneous hunter-gatherer ancestries within Late Neolithic communities, reaching up to ∼63.3% in some individuals, and variable genetic contributions of steppe herders in Bell Beaker populations. We provide an estimate as late as ∼3,800 years BCE for the admixture between Neolithic and Mesolithic populations and as early as ∼2,650 years BCE for the arrival of steppe-related ancestry. The genomic heterogeneity characterized underlines the complex history of human interactions even at the local scale.

Exploring late Paleolithic and Mesolithic diet in the Eastern Alpine region of Italy through multiple proxies.

OBJECTIVES: The analysis of prehistoric human dietary habits is key for understanding the effects of paleoenvironmental changes on the evolution of cultural and social human behaviors. In this study, we compare results from zooarchaeological, stable isotope and dental calculus analyses as well as lower second molar macrowear patterns to gain a broader understanding of the diet of three individuals who lived between the end of the Late Pleistocene and the Early Holocene (ca., 17-8 ky cal BP) in the Eastern Alpine region of Italy. MATERIALS AND METHODS: We analyze individuals buried at the sites of Riparo Tagliente (Verona), Riparo Villabruna, and Mondeval de Sora (Belluno). The three burials provide a unique dataset for diachronically exploring the influence of climatic changes on human subsistence strategies. RESULTS: Isotopic results indicate that all individuals likely relied on both terrestrial and freshwater animal proteins. Even though dental calculus analysis was, in part, hindered by the amount of mineral deposit available on the teeth, tooth macrowear study suggests that the dietary habits of the individuals included plant foods. Moreover, differences in macrowear patterns of lower second molars have been documented between Neanderthals and modern humans in the present sample, due to a prevalence of Buccal wear among the former as opposed to higher values of Lingual wear in modern human teeth. DISCUSSION: Isotopic analyses have emphasized the contribution of animal proteins in the diet of the three foragers from the Eastern Alpine region. The possible intake of carbohydrate-rich plant foods, suggested by the retrieval of plant remains in dental calculus, is supported by the signal of macrowear analysis. Moreover, the latter method indicates that the distribution of macrowear in lower second molars (M2 s) allows us to discriminate between Neanderthals and modern humans within the present reference sample. Overall, our results show these three prehistoric hunter-gatherers were well adapted to the environment in which they lived exploiting many natural resources.

Late Glacial and Early Holocene human demographic responses to climatic and environmental change in Atlantic Iberia.

Successive generations of hunter-gatherers of the Late Glacial and Early Holocene in Iberia had to contend with rapidly changing environments and climatic conditions. This constrained their economic resources and capacity for demographic growth. The Atlantic façade of Iberia was occupied throughout these times and witnessed very significant environmental transformations. Archaeology offers a perspective on how past human population ecologies changed in response to this scenario. Archaeological radiocarbon data are used here to reconstruct demographics of the region over the long term. We introduce various quantitative methods that allow us to develop palaeodemographic and spatio-temporal models of population growth and density, and compare our results to independent records of palaeoenvironmental and palaeodietary change, and growth rates derived from skeletal data. Our results demonstrate that late glacial population growth was stifled by the Younger Dryas stadial, but populations grew in size and density during the Early to Middle Holocene transition. This growth was fuelled in part by an increased dependence on marine and estuarine food sources, demonstrating how the environment was linked to demographic change via the resource base, and ultimately the carrying capacity of the environment. This article is part of the theme issue 'Cross-disciplinary approaches to prehistoric demography'.

Demographic estimates from the Palaeolithic-Mesolithic boundary in Scandinavia: comparative benchmarks and novel insights.

Prehistoric demography has recently risen to prominence as a potentially explanatory variable for episodes of cultural change as documented in the archaeological and ethnographic record. While this has resulted in a veritable boom in methodological developments seeking to address temporal changes in the relative size of prehistoric populations, little work has focused on the manner in which population dynamics manifests across a spatial dimension. Most recently, the so-called Cologne Protocol has led the way in this endeavour. However, strict requirements of raw-material exchange data as analytical inputs have prevented further applications of the protocol to regions outside of continental Europe. We apply an adjusted approach of the protocol that makes it transferable to cases in other parts of the world, while demonstrating its use by providing comparative benchmarks of previous research on the Late Glacial Final Palaeolithic of southern Scandinavia, and novel insights from the early Holocene pioneer colonization of coastal Norway. We demonstrate again that population size and densities remained fairly low throughout the Late Glacial, and well into the early Holocene. We suggest that such low population densities have played a significant role in shaping what may have been episodes of cultural loss, as well as potentially longer periods of only relatively minor degrees of cultural change. This article is part of the theme issue 'Cross-disciplinary approaches to prehistoric demography'.

Let's groove: attachment techniques of Eurasian elk (Alces alces) tooth pendants at the Late Mesolithic cemetery Yuzhniy Oleniy Ostrov (Lake Onega, Russia).

More than 4300 Eurasian elk (Alces alces) incisors, most of them pendants, were found in 84 burials in the Late Mesolithic cemetery of Yuzhniy Oleniy Ostrov, Northwest Russia. We analysed the manufacture techniques of elk teeth (4014), in the collection of the Peter the Great Museum of Anthropology and Ethnography, St Petersburg. A striking observation is that the manufacture of these pendants is similar in all burials. Teeth were worked by carving one or several grooves around the root tip. In addition to grooved ones, a number of teeth were not worked at all. The uniformity of the chosen species, tooth and techniques indicates that strict norms prevailed in the pendant industry. Despite the overall similarity, our study shows some variation in making pendants. A groove can cut the whole circumference of the root, or several distinct grooves can mark opposite sides of the root. Sometimes the grooves are deep and made carefully, and sometimes they are weak and made hastily. A typology of various groove types was created. In many graves, one groove type dominates. We interpret that this inter-burial variation and domination of one type resulted from personal choice and taste based on practicality. Such variation could also be associated with kin identifiers, but we did not find clear support for that in our study. Our study indicates that the groove types as such had no connection with particular ornaments, garments or hanging positions.

Organic residue analysis shows sub-regional patterns in the use of pottery by Northern European hunter-gatherers.

The introduction of pottery vessels to Europe has long been seen as closely linked with the spread of agriculture and pastoralism from the Near East. The adoption of pottery technology by hunter-gatherers in Northern and Eastern Europe does not fit this paradigm, and its role within these communities is so far unresolved. To investigate the motivations for hunter-gatherer pottery use, here, we present the systematic analysis of the contents of 528 early vessels from the Baltic Sea region, mostly dating to the late 6th-5th millennium cal BC, using molecular and isotopic characterization techniques. The results demonstrate clear sub-regional trends in the use of ceramics by hunter-gatherers; aquatic resources in the Eastern Baltic, non-ruminant animal fats in the Southeastern Baltic, and a more variable use, including ruminant animal products, in the Western Baltic, potentially including dairy. We found surprisingly little evidence for the use of ceramics for non-culinary activities, such as the production of resins. We attribute the emergence of these sub-regional cuisines to the diffusion of new culinary ideas afforded by the adoption of pottery, e.g. cooking and combining foods, but culturally contextualized and influenced by traditional practices.

Survival of Late Pleistocene Hunter-Gatherer Ancestry in the Iberian Peninsula.

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 [3-9]. 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. VIDEO ABSTRACT.

The Late Upper Palaeolithic and earliest Mesolithic evidence of burials in Europe.

Burials of the Late Palaeolithic (14 000-11 600 cal years before present, henceforth BP) are a rare phenomenon in Europe. Several sites possess burials of single and double individuals. As with the preceding Magdalenian, the burial of more than two individuals in the same grave cutting seems to be unusual, but does occur occasionally. The deposition of isolated and disarticulated human remains with or without cut marks seems additionally to belong to the Magdalenian context. In the final Palaeolithic phase (13 000-11 600 cal years BP) there is evidence for cemetery-like clusters of burials, which contrast to the Magdalenian evidence, instead showing some similarities with the succeeding Mesolithic. The earliest Mesolithic burials 11 600-10 500 cal BP) are a very rare phenomenon, covering a short time span between the beginning of the Preboreal and the beginning of the Boreal phase of the early Holocene. Here the evidence includes single inhumations, cemetery-like structures and a number of isolated human remains. Caves and rock shelters were the most common places for inhumations in both the final Palaeolithic and the early Mesolithic. Although the number of sites with a chronological continuity from the LUP to the Early Mesolithic burial is low, several aspects indicate a general continuity in burial patterns over this period. Apart from this continuity, the Mesolithic burials in general seem to represent a new level of diversity in burial practices.This article is part of the theme issue 'Evolutionary thanatology: impacts of the dead on the living in humans and other animals'.

Resolving relationships between several Neolithic and Mesolithic populations in Northern Eurasia using geometric morphometrics.

OBJECTIVES: Remains from several Eastern European and Siberian Mesolithic and Neolithic sites are analysed to clarify their biological relationships. We assume that groups' geographical distances correlate with genetic and, therefore, morphological distances between them. MATERIALS AND METHODS: Material includes complete male crania from several Mesolithic and Neolithic burial sites across Northern Eurasia and from several modern populations. Geometric morphometrics and multivariate statistical techniques are applied to explore morphological trends, group distances, and correlations with their geographical position, climate, and the time of origin. RESULTS: Despite an overlap in the morphology among the modern and archeological groups, some of them show significant morphological distances. Geographical parameters account for only a small proportion of cranial variation in the sample, with larger variance explained by geography and age together. Expectations of isolation by distance are met in some but not in all cases. Climate accounts for a large proportion of autocorrelation with geography. Nearest-neighbor joining trees demonstrate group relationships predicted by the regression on geography and on climate. DISCUSSION: The obtained results are discussed in application to relationships between particular groups. Unlike the Ukrainian Mesolithic, the Yuzhny Oleni Ostrov Mesolithic displays a high morphological affinity with several groups from Northern Eurasia of both European and Asian origin. A possibility of a common substrate for the Yuzhny Oleni Ostrov Mesolithic and Siberian Neolithic groups is reviewed. The Siberian Neolithic is shown to have morphological connection with both modern Siberian groups and the Native North Americans.

Paleogenomic Evidence for Multi-generational Mixing between Neolithic Farmers and Mesolithic Hunter-Gatherers in the Lower Danube Basin.

The transition from hunting and gathering to farming involved profound cultural and technological changes. In Western and Central Europe, these changes occurred rapidly and synchronously after the arrival of early farmers of Anatolian origin [1-3], who largely replaced the local Mesolithic hunter-gatherers [1, 4-6]. Further east, in the Baltic region, the transition was gradual, with little or no genetic input from incoming farmers [7]. Here we use ancient DNA to investigate the relationship between hunter-gatherers and farmers in the Lower Danube basin, a geographically intermediate area that is characterized by a rapid Neolithic transition but also by the presence of archaeological evidence that points to cultural exchange, and thus possible admixture, between hunter-gatherers and farmers. We recovered four human paleogenomes (1.1× to 4.1× coverage) from Romania spanning a time transect between 8.8 thousand years ago (kya) and 5.4 kya and supplemented them with two Mesolithic genomes (1.7× and 5.3×) from Spain to provide further context on the genetic background of Mesolithic Europe. Our results show major Western hunter-gatherer (WHG) ancestry in a Romanian Eneolithic sample with a minor, but sizeable, contribution from Anatolian farmers, suggesting multiple admixture events between hunter-gatherers and farmers. Dietary stable-isotope analysis of this sample suggests a mixed terrestrial/aquatic diet. Our results provide support for complex interactions among hunter-gatherers and farmers in the Danube basin, demonstrating that in some regions, demic and cultural diffusion were not mutually exclusive, but merely the ends of a continuum for the process of Neolithization.