A High-Coverage Mesolithic Aurochs Genome and Effective Leveraging of Ancient Cattle Genomes Using Whole Genome Imputation.

Ancient genomic analyses are often restricted to utilizing pseudohaploid data due to low genome coverage. Leveraging low-coverage data by imputation to calculate phased diploid genotypes that enables haplotype-based interrogation and single nucleotide polymorphism (SNP) calling at unsequenced positions is highly desirable. This has not been investigated for ancient cattle genomes despite these being compelling subjects for archeological, evolutionary, and economic reasons. Here, we test this approach by sequencing a Mesolithic European aurochs (18.49×; 9,852 to 9,376 calBCE) and an Early Medieval European cow (18.69×; 427 to 580 calCE) and combine these with published individuals: two ancient and three modern. We downsample these genomes (0.25×, 0.5×, 1.0×, and 2.0×) and impute diploid genotypes, utilizing a reference panel of 171 published modern cattle genomes that we curated for 21.7 million (Mn) phased SNPs. We recover high densities of correct calls with an accuracy of >99.1% at variant sites for the lowest downsample depth of 0.25×, increasing to >99.5% for 2.0× (transversions only, minor allele frequency [MAF] ≥ 2.5%). The recovery of SNPs correlates with coverage; on average, 58% of sites are recovered for 0.25× increasing to 87% for 2.0×, utilizing an average of 3.5 million (Mn) transversions (MAF ≥2.5%), even in the aurochs, despite the highest temporal distance from the modern reference panel. Our imputed genomes behave similarly to directly called data in allele frequency-based analyses, for example consistently identifying runs of homozygosity >2 Mb, including a long homozygous region in the Mesolithic European aurochs.

Long range inland-coastal networks during the Late Magdalenian: evidence for individual acquisition of marine resources at Andernach-Martinsberg, German Central Rhineland.

Recent re-examination of the osseous material assemblage from Andernach-Martinsberg, Central Rhineland, has resulted in the identification of an implement manufactured from cetacean bone (probably whale). Argued to be the proximal half of a foreshaft, this artefact is not only one of few such projectile elements to be identified in Magdalenian deposits in northern Europe, but also demonstrates that the exploitation of marine raw materials for use in manufacturing projectile elements is not restricted to southern France, instead extending to at least inland Germany. Additionally, in conjunction with the appearance of marine molluscs and engravings of seals at Andernach, it can now be forcefully argued that this region formed part of an inland-coastal network during the Late Magdalenian and allows us for the first time to suggest that we can identify the movements of individuals transporting valued marine sourced raw materials and their personal experiences across this vast region.

Nitrogen palaeo-isoscapes: Changing spatial gradients of faunal δ15N in late Pleistocene and early Holocene Europe.

Nitrogen isotope ratio analysis (δ15N) of animal tissue is widely used in archaeology and palaeoecology to investigate diet and ecological niche. Data interpretations require an understanding of nitrogen isotope compositions at the base of the food web (baseline δ15N). Significant variation in animal δ15N has been recognised at various spatiotemporal scales and related to changes both in baseline δ15N, linked to environmental and climatic influence on the terrestrial nitrogen cycle, and animal ecology. Isoscapes (models of isotope spatial variation) have proved a useful tool for investigating spatial variability in biogeochemical cycles in present-day marine and terrestrial ecosystems, but so far, their application to palaeo-data has been more limited. Here, we present time-sliced nitrogen isoscapes for late Pleistocene and early Holocene Europe (c. 50,000 to 10,000 years BP) using herbivore collagen δ15N data. This period covers the Last Glacial-Interglacial Transition, during which significant variation in the terrestrial nitrogen cycle occurred. We use generalized linear mixed modelling approaches for interpolation and test models which both include and exclude climate covariate data. Our results show clear changes in spatial gradients of δ15N through time. Prediction of the lowest faunal δ15N values in northern latitudes after, rather than during, the Last Glacial Maximum is consistent with the Late Glacial Nitrogen Excursion (LGNE). We find that including climatic covariate data does not significantly improve model performance. These findings have implications for investigating the drivers of the LGNE, which has been linked to increased landscape moisture and permafrost thaw, and for understanding changing isotopic baselines, which are fundamental for studies investigating diets, niche partitioning, and migration of higher trophic level animals.

Screening archaeological bone for palaeogenetic and palaeoproteomic studies.

The recovery and analysis of ancient DNA and protein from archaeological bone is time-consuming and expensive to carry out, while it involves the partial or complete destruction of valuable or rare specimens. The fields of palaeogenetic and palaeoproteomic research would benefit greatly from techniques that can assess the molecular quality prior to sampling. To be relevant, such screening methods should be effective, minimally-destructive, and rapid. This study reports results based on spectroscopic (Fourier-transform infrared spectroscopy in attenuated total reflectance [FTIR-ATR]; n = 266), palaeoproteomic (collagen content; n = 226), and palaeogenetic (endogenous DNA content; n = 88) techniques. We establish thresholds for three different FTIR indices, a) the infrared splitting factor [IRSF] that assesses relative changes in bioapatite crystals' size and homogeneity; b) the carbonate-to-phosphate [C/P] ratio as a relative measure of carbonate content in bioapatite crystals; and c) the amide-to-phosphate ratio [Am/P] for assessing the relative organic content preserved in bone. These thresholds are both extremely reliable and easy to apply for the successful and rapid distinction between well- and poorly-preserved specimens. This is a milestone for choosing appropriate samples prior to genomic and collagen analyses, with important implications for biomolecular archaeology and palaeontology.

Radiocarbon and stable isotope investigations at the Central Rhineland sites of Gönnersdorf and Andernach-Martinsberg, Germany.

The late glacial open-air sites of Gönnersdorf and Andernach-Martinsberg in the German Central Rhineland are well known for their Magdalenian occupation and activities. The latter site also produced evidence for a younger, Final Palaeolithic occupation of the locality by people of the Federmessergruppen. Both sites are particularly well preserved, largely due to their burial beneath volcanic deposits of the late glacial Laacher See eruption. We conducted a program of AMS radiocarbon dating and stable isotope analyses with the aim of improving understanding of the chronological history and ecological setting of the two sites. Previously published radiocarbon dates appeared to indicate that the earliest Magdalenian occupation at Gönnersdorf fell around 12,900 uncalibrated (14)C yr BP, while the earliest occupation at Andernach may have been more than 500 radiocarbon years earlier. The AMS determinations presented here revise this impression and suggest that the onset of occupation at the two sites was in fact simultaneous and prior to the warming of Greenland Interstadial GI 1e. At Gönnersdorf, a chronological hiatus exists between the main Magdalenian faunal assemblage and mega-faunal remains interpreted as collected sub-fossil material. At Andernach-Martinsberg, there is a clear chronological hiatus between the Magdalenian occupation and subsequent Federmessergruppen activities at the site. However, an intermediate radiocarbon date on an atypically preserved horse bone is suggestive of ephemeral human visits to the site between these well demonstrated phases. A date of similar age on an elk bone from Gönnersdorf may indicate broadly contemporaneous human presence at Gönnersdorf too. Stable isotope analysis of faunal remains from Gönnersdorf and Andernach-Martinsberg was conducted with the aim of both reconstructing and comparing local environmental conditions at the two sites, and also potentially identifying subtle variations in the chronological development of the two sites not detectable at the level of precision of current radiocarbon dating techniques. No spatial trends in the faunal isotope signatures were observed within each site. In the case of samples with both radiocarbon and isotope data, no chronological pattern was observed for the isotope results. The Magdalenian faunal isotope signatures at the two sites resembled each other, suggesting comparable climatic and environmental conditions. The faunal delta(13)C signatures at Gönnersdorf and Andernach-Martinsberg were similar to those at contemporary European sites. While the faunal delta(15)N values were similar to those at contemporary sites in Germany, the UK, and Belgium, they were lower than those from the South of France. This difference in delta(15)N values is thought to relate to regional differences in the timing of changes in soil and plant nitrogen cycling in response to ameliorating climatic conditions.

Stable isotopes reveal patterns of diet and mobility in the last Neandertals and first modern humans in Europe.

Correlating cultural, technological and ecological aspects of both Upper Pleistocene modern humans (UPMHs) and Neandertals provides a useful approach for achieving robust predictions about what makes us human. Here we present ecological information for a period of special relevance in human evolution, the time of replacement of Neandertals by modern humans during the Late Pleistocene in Europe. Using the stable isotopic approach, we shed light on aspects of diet and mobility of the late Neandertals and UPMHs from the cave sites of the Troisième caverne of Goyet and Spy in Belgium. We demonstrate that their diet was essentially similar, relying on the same terrestrial herbivores, whereas mobility strategies indicate considerable differences between Neandertal groups, as well as in comparison to UPMHs. Our results indicate that UPMHs exploited their environment to a greater extent than Neandertals and support the hypothesis that UPMHs had a substantial impact not only on the population dynamics of large mammals but also on the whole structure of the ecosystem since their initial arrival in Europe.

Evidence for close-range hunting by last interglacial Neanderthals.

Animal resources have been part of hominin diets since around 2.5 million years ago, with sharp-edged stone tools facilitating access to carcasses. How exactly hominins acquired animal prey and how hunting strategies varied through time and space is far from clear. The oldest possible hunting weapons known from the archaeological record are 300,000 to 400,000-year-old sharpened wooden staves. These may have been used as throwing and/or close-range thrusting spears, but actual data on how such objects were used are lacking, as unambiguous lesions caused by such weapon-like objects are unknown for most of human prehistory. Here, we report perforations observed on two fallow deer skeletons from Neumark-Nord, Germany, retrieved during excavations of 120,000-year-old lake shore deposits with abundant traces of Neanderthal presence. Detailed studies of the perforations, including micro-computed tomography imaging and ballistic experiments, demonstrate that they resulted from the close-range use of thrusting spears. Such confrontational ways of hunting require close cooperation between participants, and over time may have shaped important aspects of hominin biology and behaviour.

At the end of the 14C time scale--the Middle to Upper Paleolithic record of western Eurasia.

The dynamics of change underlying the demographic processes that led to the replacement of Neandertals by Anatomically Modern Humans (AMH) and the emergence of what are recognized as Upper Paleolithic technologies and behavior can only be understood with reference to the underlying chronological framework. This paper examines the European chronometric (mainly radiocarbon-based) record for the period between ca. 40 and 30 ka 14C BP and proposes a relatively rapid transition within some 2,500 years. This can be summarized in the following falsifiable hypotheses: (1) final Middle Paleolithic (FMP) "transitional" industries (Uluzzian, Chatelperronian, leaf-point industries) were made by Neandertals and date predominantly to between ca. 41 and 38 ka 14C BP, but not younger than 35/34 ka 14C BP; (2) initial (IUP) and early (EUP) Upper Paleolithic "transitional" industries (Bachokirian, Bohunician, Protoaurignacian, Kostenki 14) will date to between ca. 39/38 and 35 ka 14C BP and document the appearance of AMH in Europe; (3) the earliest Aurignacian (I) appears throughout Europe quasi simultaneously at ca. 35 ka 14C BP. The earliest appearance of figurative art is documented only for a later phase ca. 33.0/32.5-29.2 ka 14C BP. Taken together, the Middle to Upper Paleolithic transition appears to be a cumulative process involving the acquisition of different elements of "behavioral modernity" through several "stages of innovation."

A critical review of the German Paleolithic hominin record.

We review the hominin fossil record from western Central Europe in light of the recent major revisions of the geochronological context. The mandible from Mauer (Homo heidelbergensis), dated to circa 500,000 years ago, continues to represent the earliest German hominin and may coincide with the occupation of Europe north of the high alpine mountain chains. Only limited new evidence is available for the Middle Pleistocene, mostly in the form of skull fragments, a pattern that may relate to taphonomic processes. These finds and their ages suggest the gradual evolution of a suite of Neandertal features during this period. Despite new finds of classic Neandertals, there is no clear proof for Neandertal burial from Germany. Alternatively, cut marks on a skull fragment from the Neandertal type site suggest special treatment of that individual. New Accelerator Mass Spectrometry (AMS) radiocarbon dates of previous finds leave little reliably dated evidence for anatomically modern humans (AMH) in Europe before 30,000 BP; the remains from Hahnöfersand, Binshof-Speyer, Paderborn-Sande, and Vogelherd are now of Holocene age. Thus, a correlation of AMH with the Aurignacian remains to be proven, and the general idea of a long coexistence of Neandertals and AMH in Europe may be questioned. In western Central Europe, evidence of Gravettian human fossils is also very limited, although a new double grave from lower Austria may be relevant. The only dated burial from the German Upper Paleolithic (from Mittlere Klause) falls into a time period (circa 18,600 BP) represented by only a few occupation sites in western Central Europe. A number of human remains at Magdalenian sites appear to result from variable (secondary) burial practices. In contrast, the Final Paleolithic (circa 12,000-9600 cal. BC) yields an increase of hominin finds, including multiple burials (Bonn-Oberkassel, Neuwied-Irlich), similar to the situation in western and southern Europe.