Reconstructing how prehistoric humans used the products obtained from large cetaceans is challenging, but key to understand the history of early human coastal adaptations. Here we report the multiproxy analysis (ZooMS, radiocarbon, stable isotopes) of worked objects made of whale bone, and unworked whale bone fragments, found at Upper Paleolithic sites (Magdalenian) around the Bay of Biscay. Taxonomic identification using ZooMS reveals at least five species of large whales, expanding the range of known taxa whose products were utilized by humans in this period. Radiocarbon places the use of whale products ca. 20–14 ka cal BP, with a maximum diffusion and diversity at 17.5–16 ka cal BP, making it the oldest evidence of whale-bone working to our knowledge. δ¹³C and δ¹⁵N stable isotope values reflect taxon-specific differences in foraging behavior. The diversity and chronology of these cetacean populations attest to the richness of the marine ecosystem of the Bay of Biscay in the late Paleolithic, broadening our understanding of coastal adaptations at that time.

Archaeological evidence suggests that dogs diverged from wolves during the Palaeolithic, more than 15,000 years ago ^1–7 . The earliest unequivocal genetic evidence, however, is associated with dog remains from Mesolithic archaeological contexts approximately 10,900 years ago ^8,9 . Here we generate both nuclear and mitochondrial genomes from canid remains at Pınarbaşı in Türkiye (15,800 years ago) ¹⁰ and Gough’s Cave in the UK (14,300 years ago) ¹¹ , as well as from dogs excavated from two Mesolithic sites in Serbia (Padina between 11,500–7,900 years ago and Vlasac 8,900 years ago) ^12,13 . Our analyses indicate that a genetically homogeneous dog population was already widely distributed across Europe and Anatolia during the Late Upper Palaeolithic (by at least 14,300 years ago). This finding suggests that dogs were exchanged among genetically and culturally distinct western Eurasian Late Palaeolithic human populations, namely the Magdalenian, Epigravettian and Anatolian hunter-gatherers ^10,14–16 . Last, we identify a major influx of eastern Eurasian dog ancestry during the Mesolithic, concomitant with the movement of eastern hunter-gatherer populations into Europe ¹⁴ , which led to the establishment of the primary ancestry characteristics that define European dog populations today.

Collagen glue has been used for nearly two centuries to consolidate bone material, although its prevalence in museum collections is only now becoming visible. Identifying and removing collagen glue is crucial before the execution of any geochemical or molecular analyses. Palaeolithic bone objects from old excavations intended for radiocarbon dating were first analysed using ZooMS (Zooarchaeology by Mass Spectrometry) to identify the animal species, however peaks characteristic of both cattle and whale were discovered. Two extraction methods for ZooMS were tested to identify the authentic animal species of these objects, which revealed that these were originally whale bone objects that had been consolidated with cattle collagen glue. This is the first time animal collagen glue has been identified in archaeological remains with ZooMS, illustrating again the incredible versatility of this technique. Another technique, Fourier Transform Infrared Spectroscopy in Attenuated Total Reflectance mode (FTIR-ATR), was also tested if it could rapidly identify the presence of collagen glue in archaeological bone material, which was not the case. Two other cleaning methods were tested to remove bone glue contamination prior to radiocarbon dating, along with two modified collagen extraction methods for ZooMS. These methods were applied to bone blank samples (FmC = 0.0031 ± 0.0002, (n = 219), 47 336 ± 277 yr BP) that were experimentally consolidated with collagen glue and to the Palaeolithic bone material (ca. 15 000 and 12 000 yr BP). The experimental bone blanks produced excellent¹⁴C ages, suggesting the cleaning methods were successful, however the¹⁴C ages for some of the Palaeolithic material remained too young considering their contextual age, suggesting that the collagen glue contamination had most likely cross-linked to the authentic collagen molecule. More research is needed in order to gain a deeper understanding of the occurrence and elimination of cross-linked collagen-based glues in material from museum collections.

Museum collections are extremely valuable sources of material for ongoing research, although the conservation history of some objects is not always recorded, which can be problematic for chemical analyses. While most contamination is removed using the acid-base-acid treatment, this may not be the case for cross-linked contamination. The XAD resin protocol was implemented at the radiocarbon (¹⁴C) laboratory in the Muséum national d’Histoire naturelle, and the setup was tested using known age bone samples and a consolidated Palaeolithic bone. Known age samples were consolidated with shellac or Paraloid, aged for a month, treated with or without the XAD resin and¹⁴C dated. Bone blank results showed that XAD resin was able to remove shellac, which was not the case for the ABA-only method. Results from VIRI I were more variable and VIRI F was possibly too young to show the effects of the consolidants. Two¹⁴C dates on the Palaeolithic bone after XAD treatment are statistically the same, while a sample without XAD treatment was significantly older, suggesting that the contaminant was not fully removed by the ABA-only treatment. This study demonstrates the potential of the XAD treatment to clean heritage bone samples stored in museums prior to geochemical analyses.