Oxygen isotopes in bioarchaeology : Principles and applications, challenges and opportunities

The Max-Planck-Society and the University of Aberdeen are thanked for professional and financial support during production of this manuscript. Ciara Gigleux of CG editing is thanked for help with copy-editing. Thanks are also due to Matthew Collins (Copenhagen/Cambridge) and Michelle Alexander (York) for contributions to the production of Figure 8. We also thank the Editors and two anonymous reviewers whose comments on an earlier version of this manuscript greatly improved this work.Peer reviewedPostprin

Anion exchange resin and slow precipitation preclude the need for pretreatments in silver phosphate preparation for oxygen isotope analysis of bioapatites

The authors would like to thank Wolfram Meier-Augenstein (Robert Gordon University) for advice on TC/EA677 IRMS and to Raquel Maria (Kimmel Center for Archaeological Science, Weizmann Institute of Science) for advice on FTIR-ATR. Thanks to Birke Brumme (MPI EVA) for practical support with sample preparation. Thanks are also due to Sahra Talamo (MPI EVA/University of Bologna) for providing aliquots of the S-EVA-2000 and S-EVA-2001 in-house bone standards and to Klervia Jaouen (MPI EVA/Géosciences Environnment Toulouse) for providing extracted collagen used in the preparation of synthetic bones. This research was funded by the Max-Planck-Society as part of SP’s doctoral research. The authors would also like to thank the Max-Planck-Society, the University of Aberdeen and the Vreije Universiteit Brussels for professional and financial support during the production of this manuscript. CS thanks the Research Foundation - Flanders for his post-doctoral fellowship. We also thank Christophe Lécuyer and an anonymous reviewer for their valuable comments and suggestions.Peer reviewedPostprin

Late Pleistocene Neanderthal exploitation of stable and mosaic ecosystems in northern Iberia shown by multi-isotope evidence

The carbon, nitrogen and sulphur stable isotope analyses and the stable isotope analyses of bioapatite carbonates were funded as part of the ABRUPT project (HAR2017-84997-P) funded by the Ministry of Science, Innovation and Universities and the SUBSILIENCE project funded by the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement no. 818299—ERC-2018-Consolidator), both awarded to ABM-A. SP was supported by the Max Planck Society and the University of Aberdeen during the time of this project, and the oxygen isotope analysis of bioapatite phosphates was funded by the Max Planck Society. Access to the archaeological collections was granted by the Museo de Arqueología de Bizkaia (Basque Government), and initial sampling for carbon and nitrogen isotope analysis was achieved by Hazel Reade funded by the FP7-PEOPLE-2012-CIG- 322112 project) and ABM-A. We appreciate Joseba Rios-Garaizar's advice about Axlor stratigraphy during the sampling process. We thank Ignacio Valera (IBBTEC, University of Cantabria) for kindly allowing the use of his laboratory facilities for collagen extraction. We thank Carlos Revilla Gómez (IBBTEC, University of Cantabria) for laboratory assistance during collagen extraction. Thanks are also due to Manuel Trost (MPI-EVA) for assistance during silver phosphate preparation and to Sven Steinbrenner for assistance with TC/EA-IRMS. KB is supported by a Philip Leverhulme Prize from the Leverhulme Trust (PLP-2019-284).Peer reviewedPublisher PD

Oxygen isotope analyses of Equus teeth evidences early Eemian and early Weichselian palaeotemperatures at the Middle Palaeolithic site of Neumark-Nord 2, Saxony-Anhalt, Germany

We thank Annabell Reiner (MPI-EVA) for technical and practical support with preparation of samples and Bernd Steinhilber for the oxygen isotope measurements of the silver phosphate samples at the Institut für Geowissenschaften (Universität Tübingen); Thanks to the Landesamt für Denkmalpflege und Archäologie, Sachsen-Anhalt, and Landesmuseum Sachsen-Anhalt in Halle for providing samples; and to Geoff Smith (RGZM Monrepos and MPI-EVA) for comments on earlier versions of this manuscript. Financial support for the Neumark-Nord 2 excavations was provided by the Lausitzer Mitteldeutsche Braunkohlengesellschaft mbH, the Landesamt für Denkmalpflege und Archäologie Sachsen-Anhalt (Harald Meller, Susanne Friederich), the Römisch-Germanisches Zentralmuseum Mainz, the Leids Universiteits Fonds “Campagne voor Leiden” program and the NetherlandsOrganization for Scientific Research (N.W.O.). The isotope research was funded by the Max Planck Institute and a Deutscher Akademischer Austausch Dienst Junior Research Grant to KB (ref: A0970923). Thanks also to the University of Aberdeen, and The Leverhulme Trust (RPG-2017-410) for financial and professional support during this project and preparation of the manuscript. TT acknowledges funding by the German National Science foundation in the framework of the Emmy Noether Program (DFG grant TU 148/2-1 “Bone Geochemistry”).Peer reviewedPostprin

Ecological evolution in northern Iberia (SW Europe) during the Late Pleistocene through isotopic analysis on ungulate teeth.

During the Late Pleistocene, stadial and interstadial fluctuations affected vegetation, fauna, and human groups that were forced to cope with these pronounced climatic and environmental changes in time and space. These changes were especially abrupt during the Marine Isotopic Stage (MIS) 3. However, little is still known about the local and regional climatic conditions experienced by hominins in Europe. Here we reconstruct the climatic trends in northern Iberia considering the stable isotopic composition of ungulate skeletal tissues found in archaeological deposits dated between 80 to 15,000 cal BP. The carbon and oxygen isotopic composition preserved in the carbonate fraction of tooth enamel provides a reliable and high-resolution proxy of the food and water consumed by these animals, which is indirectly related to the local vegetation, environment, and climate, allowing us to estimate paleotemperatures and rainfall data. This study presents 44 bovine, equid, and cervid teeth from five archaeological sites in the Vasco-Cantabrian region (El Castillo, El Otero, Axlor, Labeko Koba, Aitzbitarte III) and one in the Mediterranean area (Canyars), where human evidence is attested from the Mousterian to the Magdalenian. The carbon isotope values reflect animals feeding on C3 plants with a mix-feeder diet mainly developed in open environments. However, carbon isotope value ranges point to differentiated ecological niches for equids and bovines, especially during the Aurignacian in the Vasco-Cantabrian region. Temperature estimations based on oxygen isotopic compositions and rainfall obtained from carbon isotopic compositions indicate colder and more arid conditions than nowadays from the Late Mousterian to the Aurignacian. The contemporary Mediterranean site shows slightly lower temperatures related to an arid period when animals mainly graze in open landscapes. In the Vasco-Cantabrian region, during the MIS2, the Gravettian data reflect a landscape opening, whereas the Magdalenian point to warmer conditions but still arid

Reconstructing Late Pleistocene paleoclimate at the scale of human behavior: an example from the Neandertal occupation of La Ferrassie (France)

Exploring the role of changing climates in human evolution is currently impeded by a scarcity of climatic information at the same temporal scale as the human behaviors documented in archaeological sites. This is mainly caused by high uncertainties in the chronometric dates used to correlate long-term climatic records with archaeological deposits. One solution is to generate climatic data directly from archaeological materials representing human behavior. Here we use oxygen isotope measurements of Bos/Bison tooth enamel to reconstruct summer and winter temperatures in the Late Pleistocene when Neandertals were using the site of La Ferrassie. Our results indicate that, despite the generally cold conditions of the broader period and despite direct evidence for cold features in certain sediments at the site, Neandertals used the site predominantly when climatic conditions were mild, similar to conditions in modern day France. We suggest that due to millennial scale climate variability, the periods of human activity and their climatic characteristics may not be representative of average conditions inferred from chronological correlations with long-term climatic records. These results highlight the importance of using direct routes, such as the high-resolution archives in tooth enamel from anthropogenically accumulated faunal assemblages, to establish climatic conditions at a human scale.Projekt DEALinfo:eu-repo/semantics/publishedVersio

Subarctic climate for the earliest Homo sapiens in Europe

Acknowledgments The re-excavation of Bacho Kiro Cave was jointly conducted by the National Institute of Archaeology with Museum, Bulgarian Academy of Sciences, Sofia and the Department of Human Evolution at the MPI-EVA. We would like to thank the National Museum of Natural History (Sofia), the Archaeology Department at the New Bulgarian University (Sofia), the Regional Museum of History in Gabrovo, and the History Museum in Dryanovo for assistance on this project and the opportunity to study the Bacho Kiro Cave faunal material. We would like to thank M. Trost, S. Hesse, M. Kaniecki, and P. Dittmann (MPI-EVA) for technical assistance during stable isotope sample preparation. S. Steinbrenner is thanked for technical assistance with TC/EA-IRMS maintenance. Thanks are also due to H. Temming and U. Schwarz (MPI-EVA) for the production of microCT scans and replicas of the sample materials. We would also like to acknowledge the assistance of to D. Veres with taking OSL samples. Last but not least we would like to thank the handling editor, S. Ortman, as well as three anonymous reviewers for their thoughtful comments that greatly improved this manuscript. Funding: The field work was financed by the Max Planck Society. The stable isotope work was funded by the Max Planck Society as part of S.P.’s doctoral project. S.P. was supported by the Max Planck Society and the University of Aberdeen. K.B. was supported by a Philip Leverhulme Prize from The Leverhulme Trust (PLP-2019-284). N.B.’s work was supported as part of a grant by the German Research Foundation (“PALÄODIET” Project 378496604). V.A. was supported by a grant from the Foundation for Science and Technology, Portugal (IF/01157/2015/CP1308/CT0002). Author contributions: The study was devised by S.P., K.B., S.P.M., J.-J.H., and T.T. Archaeological excavation was undertaken by N.S. and T.T. in collaboration with Z.R. and S.P.M. who all contributed contextual information. V.A. collected sedimentological data at the site and untertook micromorphological investigations that provided information on site formation for this study. Zooarchaeological and paleontological analyses were performed by G.M.S. and R.S. OSL dating was carried out by T.L. Radiocarbon dating and recalibration of radiocarbon dates were conducted by H.F. MC-ICPMS analysis was conducted by N.B. and S.P. Sampling, sample processing for oxygen and strontium stable isotope analysis, and TC/EA-IRMS analysis were carried out by S.P. Code and data analyses were written and conducted by S.P. N.-H.T. consulted on statistical analysis and coding. S.P. wrote the paper with input from all authors. Competing interests: The authors declare that they have no competing interests. Data and materials availability: All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials.Peer reviewedPublisher PD

Trophic position of Otodus megalodon and great white sharks through time revealed by zinc isotopes

Diet is a crucial trait of an animal’s lifestyle and ecology. The trophic level of an organism indicates its functional position within an ecosystem and holds significance for its ecology and evolution. Here, we demonstrate the use of zinc isotopes (δ66Zn) to geochemically assess the trophic level in diverse extant and extinct sharks, including the Neogene megatooth shark (Otodus megalodon) and the great white shark (Carcharodon carcharias). We reveal that dietary δ66Zn signatures are preserved in fossil shark tooth enameloid over deep geologic time and are robust recorders of each species’ trophic level. We observe significant δ66Zn differences among the Otodus and Carcharodon populations implying dietary shifts throughout the Neogene in both genera. Notably, Early Pliocene sympatric C. carcharias and O. megalodon appear to have occupied a similar mean trophic level, a finding that may hold clues to the extinction of the gigantic Neogene megatooth shark.publishedVersio

The ecology, subsistence and diet of ~45,000-year-old Homo sapiens at Ilsenhöhle in Ranis, Germany

Recent excavations at Ranis (Germany) identified an early dispersal of Homo sapiens into the higher latitudes of Europe by 45,000 years ago. Here we integrate results from zooarchaeology, palaeoproteomics, sediment DNA and stable isotopes to characterize the ecology, subsistence and diet of these early H. sapiens. We assessed all bone remains (n = 1,754) from the 2016-2022 excavations through morphology (n = 1,218) or palaeoproteomics (zooarchaeology by mass spectrometry (n = 536) and species by proteome investigation (n = 212)). Dominant taxa include reindeer, cave bear, woolly rhinoceros and horse, indicating cold climatic conditions. Numerous carnivore modifications, alongside sparse cut-marked and burnt bones, illustrate a predominant use of the site by hibernating cave bears and denning hyaenas, coupled with a fluctuating human presence. Faunal diversity and high carnivore input were further supported by ancient mammalian DNA recovered from 26 sediment samples. Bulk collagen carbon and nitrogen stable isotope data from 52 animal and 10 human remains confirm a cold steppe/tundra setting and indicate a homogenous human diet based on large terrestrial mammals. This lower-density archaeological signature matches other Lincombian-Ranisian-Jerzmanowician sites and is best explained by expedient visits of short duration by small, mobile groups of pioneer H. sapiens. [Abstract copyright: © 2024. The Author(s).

Stable isotopes show Homo sapiens dispersed into cold steppes ~45,000 years ago at Ilsenhöhle in Ranis, Germany

The spread of Homo sapiens into new habitats across Eurasia ~45,000 years ago and the concurrent disappearance of Neanderthals represents a critical evolutionary turnover in our species' history. 'Transitional' technocomplexes, such as the Lincombian-Ranisian-Jerzmanowician (LRJ), characterize the European record during this period but their makers and evolutionary significance have long remained unclear. New evidence from Ilsenhöhle in Ranis, Germany, now provides a secure connection of the LRJ to H. sapiens remains dated to ~45,000 years ago, making it one of the earliest forays of our species to central Europe. Using many stable isotope records of climate produced from 16 serially sampled equid teeth spanning ~12,500 years of LRJ and Upper Palaeolithic human occupation at Ranis, we review the ability of early humans to adapt to different climate and habitat conditions. Results show that cold climates prevailed across LRJ occupations, with a temperature decrease culminating in a pronounced cold excursion at ~45,000-43,000 cal BP. Directly dated H. sapiens remains confirm that humans used the site even during this very cold phase. Together with recent evidence from the Initial Upper Palaeolithic, this demonstrates that humans operated in severe cold conditions during many distinct early dispersals into Europe and suggests pronounced adaptability. [Abstract copyright: © 2024. The Author(s).