Identification of new Neandertal remains at Goyet and their biogeochemical characterization

The reanalysis of the Goyet material comprised (i) the revision of the human skeletal material, (ii) systematic sorting of the faunal collections to check for unidentified human remains (Supplementary Fig. S2), and (iii) a multidisciplinary study of the human remains and their context. Two-hundred and eighty three human remains were identified from different periods, including 96 bone specimens and three isolated teeth identifiable as Neandertal (Supplementary Table S1 and Supplementary Notes S3, S4 and S5). A good number (n = 47) of the bone specimens refit, reducing the total number of isolated Neandertal remains to 64 (Fig. 1 and Supplementary Table S2), of which 10 were directly radiocarbon (14C) dated, 15 were sampled for stable isotope analyses, and 10 for DNA extraction (Table 1 and Supplementary Table S3). Based on their morphology and morphometric characteristics, developmental stage and side for paired elements, as well as the successful recovery of endogenous mitochondrial DNA (mtDNA) sequences, the minimum number of individuals (MNI) represented by the Goyet sample is estimated at five (four adolescents/adults and one child represented by a single tooth; Supplementary Note S5 and Supplementary Fig. S3). Although the Neandertal sample includes cranial and postcranial elements (Fig. 1), with long bones best represented and extremities mostly absent, the minimum number of elements (MNE = 35) demonstrates a very low overall skeletal representation. The best represented elements are, in decreasing order, the tibia (six of the eight tibias expected for four adolescents/adults, 75% representation), femur and cranium (50%), humerus and mandible (25%; Supplementary Table S4).

Figure 1: Neandertal remains from the Troisième caverne of Goyet (Belgium). *Designates the specimens that have been directly dated. Scale = 3 cm. Full size image

Table 1 Sample information and results of the 14C and genetic analyses of the Neandertal remains from Goyet. Full size table

Chemical elemental analyses performed together with stable isotope analyses were used to assess collagen preservation in preparation of 14C dating (see Methods). The ecology of the Goyet Neandertals was also investigated using δ13C and δ15N isotope composition of bone collagen28. Direct 14C dates obtained from the newly identified skeletal material place the Goyet Neandertals to ca. 40.5–45.5 ky calBP. However, when the youngest ages, which likely reflect undetected bone collagen contamination, are excluded (Supplementary Note S6), we cannot rule out the possibility that the Goyet Neandertals represent a single chronological group dating to ca. 44–45.5 ky calBP. Although this appears the most parsimonious hypothesis when individual bone associations, taphonomic aspects and similar anthropogenic modifications observed across the sample are taken into account, we retain the conservative range of ca. 40.5–45.5 ky calBP for the Goyet Neandertals in the absence of definitive evidence.

Out of the 10 samples processed for genetic analysis, seven show three distinct complete or almost complete mtDNA lineages (noted 1–3 in Table 1). The newly reconstructed mtDNAs from Goyet were compared with the mtDNA of 54 modern humans, eight previously sequenced Neandertals and one Denisovan individual29,30,31,32,33,34. Phylogenetic relationships were assessed using maximum parsimony and maximum likelihood trees (Fig. 2 and Supplementary Fig. S4), confirming the analysed specimens to fall within the known diversity of Neandertal mtDNA. The Goyet Neandertal mtDNAs appear most closely related to late Neandertal mtDNAs from Central and Western Europe, such as those from the Neandertal type-site (Germany), El Sidrón (Spain) and Vindija (Croatia), which all show only modest genetic variation despite large geographic distances when compared to modern humans. As previously suggested31, this might reflect a low effective population size of Neandertals in general, and for the late Neandertals in particular.

Figure 2: Maximum parsimony tree for the seven analysed Goyet samples that produced complete or almost complete mitochondrial genomes compared to 63 published modern human, Neandertal and Denisovan mtDNAs. Numbers at the main branch nodes represent bootstrap values after 1,000 iterations. Full size image

Taphonomic analysis of the Goyet Neandertal material and anthropogenic modifications

Overall, the Neandertal remains are highly fragmented. Forty-nine percent of the bone specimens (47 out of 96) were refit to at least one other, with the number of specimens per refit set ranging from 2 to 8 (tibia I; Supplementary Fig. S5). Several examples of refits between levels 1 through 3 were also identified. None of the Neandertal bones are complete, although the proximal extremity of a hand phalanx (2878–37) is only slightly eroded (Fig. 1). Cortical surfaces are well preserved and exhibit limited post-depositional modifications. Most long bones fractures involve green breaks, as indicated by smooth margins and spiral fractures35. Traces of peeling may also provide evidence for the fresh bone fracture of a cranial fragment and several ribs (ref. 11; Supplementary Fig. S6). Although bears can produce such traces36, the presence of cutmarks on several ribs (see below) suggests that the most parsimonious hypothesis is that they are anthropogenic. Traces of human chewing37,38 are also suspected on the Neandertal phalanges but are inconclusive (Supplementary Fig. S6). The numerous unambiguous anthropogenic marks on the Goyet Neandertal remains can be attributed to three categories of bone surface modifications (Figs 3, 4, 5, Table 2, and Supplementary Figs S7 and S8):

1 Cutmarks. Nearly a third of the Neandertal specimens bear cutmarks. The locations of the limited number of cutmarks observed on the upper limb may indicate disarticulation whereas those on the lower limb are consistent with defleshing. Several cutmarks on the internal and external surfaces of the ribs may be connected to evisceration, dismemberment of the thoracic cage and removal of the thoracic muscles. An additional cutmark on the medial side of the mandible, close to the mandibular condyle, appears consistent with dismemberment. 2 Two types of percussion marks (notches and pits) were identified. Observed only on a single radius alongside several femurs and tibias, notches are likely connected to the fracturing of fresh diaphyses and marrow extraction. Percussion pits are common and probably indicate failed attempts at fracturing bones. Both percussion notches and pits were also identified on eight bones (e.g. femur I, Fig. 5). 3 Retouching marks. These marks, found on a femur and three tibias (Supplementary Figs S9–S12), result from retouching the edges of stone tools. The fact that none of the affected areas overlap on adjacent fragments suggests the bones to probably have first been marrow cracked. Femur III shows two retouching zones on the anterior and postero-medial surfaces, both located at mid-shaft. Interestingly, the traces found on the tibias are located in the same areas of the shaft on all three bones (posterior or postero-medial surface at mid-shaft). The retouchers are made on four different Neandertal bones that represent at least three of the four adolescent/adult Neandertal individuals (Supplementary Note S5).

Figure 3: Overview of the anthropogenic modifications observed on the Neandertal remains from the Troisième caverne of Goyet (Belgium). See Supplementary Fig. S8 for individual Neandertal bones with anthropogenic modifications. Skeleton diagrams modified from https://en.wikipedia.org/wiki/File:Human_skeleton_front_en.svg and https://en.wikipedia.org/wiki/File:Human_skeleton_back_en.svg using Adobe Illustrator CS4 v. 14.0.0. Full size image

Figure 4: Retouching marks (b1,b2) and cutmarks (c1,c2) present on the Goyet Neandertal bones (example of femur III). (a) femur III in anterior view; (b1,c1) close-up photos; (b2,c2) images obtained using a minidome (see Methods). Full size image

Figure 5: Percussion pits (b1,b2) and percussion notch (c1,c2) present on the Goyet Neandertal bones (example of femur I). (a) femur I in posterior view; (b1,c1) close-up photos; (b2,c2) images obtained using a minidome (see Methods). Full size image

Table 2 Numbers and proportions of Neandertal, horse, reindeer and carnivore remains bearing anthropogenic modifications and toothmarks in the Goyet assemblage. Full size table

While animal bone retouchers are common in European Middle Palaeolithic contexts (e.g., refs 39, 40, 41), Goyet is one of only four sites (Krapina in Croatia42, La Quina and Les Pradelles in France43,16) to have yielded retouchers on Neandertal skeletal elements and the sole to have produced multiple examples (Table 3). At Krapina and Les Pradelles, femur shaft fragments were used as retouchers, whereas the La Quina example is on a parietal fragment. According to the criteria proposed by Mallye et al.40, the blanks used for the Goyet retouchers made on Neandertal bones were most likely green due to the absence of scaled areas, and in addition, two of the five retoucher areas exhibit concentrated and superposed marks which imply prolonged use. The rectilinear morphology of the marks also supports the use of the bones for retouching flint flakes, the most common raw material found at Goyet.

Table 3 Description of the Neandertal bone retouchers from Goyet using the criteria of Mallye et al. 40 and Daujeard et al. 41. Full size table

Comparative taphonomic analysis of the fauna from the Troisième caverne

Due to the large size of the Goyet faunal collection (>30,000 specimens), only a sample from Dupont’s excavation was examined (see Methods; Supplementary Fig. S2 and Supplementary Table S5). The skeletal material analysed corresponds mostly to long bone shaft fragments from various species that were mixed together within the collection and did not appear to have been previously sorted. We focused on remains from levels 3 and 2, which yielded the Neandertal remains, and on material from the same storage trays containing the human remains in order to have an overview of the associated faunal spectrum and assess food procurement and management strategies. Horse and reindeer are by far the most frequent species in the studied assemblage (86% of the 1,556 identified specimens; Supplementary Table S5). No rodent toothmarks were observed, carnivore remains are relatively sparse and carnivore damage is extremely rare on the Neandertal, horse and reindeer remains (Table 2), indicating carnivores to have had limited access to the bone material.

Anatomical profiles reveal numerous similarities between the Neandertal sample on one hand and horse and reindeer on the other (Supplementary Table S6 and Supplementary Fig. S13). The tibia is the most abundant element of all three species, whereas the axial skeleton and extremities of the forelimb and hindlimb are poorly represented. Bones of the hindlimb are better represented for all three species compared to forelimb elements, this is especially the case with the Neandertal material. The only notable difference between the faunal and Neandertal remains is the high representation of cranial elements for the latter. Unfortunately, the absence of contextual data precludes an analysis of the spatial distribution of both the faunal and Neandertal remains within the Troisième caverne.

The most intensely processed Neandertal elements are femurs and tibias (Supplementary Fig. S7), which are also the bones with the highest nutritional content (meat and marrow). The same pattern was documented for horse and reindeer bones. Overall, anthropogenic marks on the Neandertal remains match those most commonly recorded on the faunal material (Supplementary Figs S14–S16). All three taxa were intensively exploited, exhibiting evidence of skinning, filleting, disarticulation and marrow extraction. However, the Neandertal remains stand out as they show a high number of percussion pits (Table 2), which may be linked to the thick cortical structure of Neandertal long bones. Although the Neandertal remains show no traces of burning, the possibility that they may have been roasted or boiled cannot be excluded. The high number of cutmarks and the fact that DNA could be successfully extracted are, however, inconsistent with this possibility44,45,46. Lastly, similar to what has been noted at other sites40,41,47, the Neandertal retouchers are made on fragments of dense bones with comparable mechanical properties to the horse and reindeer bones. At Goyet, as at several French Middle Palaeolithic sites, large bone fragments of medium and large-sized animals were selected40,41,48,49,50,51. Among the Goyet Neandertal material, the largest and thickest fragments were also selected, as was the case at Les Pradelles16 and Krapina42. Interestingly, a femur and tibias of cave bears were also among the retoucher blanks selected by Neandertals at Scladina52.

The observed patterns of faunal exploitation can be interpreted as the selective transport of meat and marrow rich elements to the site that were subsequently intensively processed. However, this apparent pattern may reflect a collection bias favoring the largest and most easily identifiable fragments. Similarities in anthropogenic marks observed on the Neandertal, horse and reindeer bones do, however, suggest similar processing and consumption patterns for all three species.