Initial Upper Palaeolithic Homo sapiens from Bacho Kiro Cave, Bulgaria


The Middle to Upper Palaeolithic transition in Europe witnessed the replacement and partial absorption of local Neanderthal populations by Homo sapiens populations of African origin1. However, this process probably varied across regions and its details remain largely unknown. In particular, the duration of chronological overlap between the two groups is much debated, as are the implications of this overlap for the nature of the biological and cultural interactions between Neanderthals and H. sapiens. Here we report the discovery and direct dating of human remains found in association with Initial Upper Palaeolithic artefacts2, from excavations at Bacho Kiro Cave (Bulgaria). Morphological analysis of a tooth and mitochondrial DNA from several hominin bone fragments, identified through proteomic screening, assign these finds to H. sapiens and link the expansion of Initial Upper Palaeolithic technologies with the spread of H. sapiens into the mid-latitudes of Eurasia before 45 thousand years ago3. The excavations yielded a wealth of bone artefacts, including pendants manufactured from cave bear teeth that are reminiscent of those later produced by the last Neanderthals of western Europe4,5,6. These finds are consistent with models based on the arrival of multiple waves of H. sapiens into Europe coming into contact with declining Neanderthal populations7,8.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1: Direct dates for hominins of the Middle to Upper Palaeolithic transition in Eurasia.
Fig. 2: Maximum parsimony tree.
Fig. 3: Bone tools and personal ornaments from Bacho Kiro Cave layers I and J (Niche 1 and Main sectors).

Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request. Genetic sequence reads from all libraries and corresponding negative controls are deposited at European Nucleotide Archive under the study accession number PRJEB35466. The FASTA files of the mitochondrial genomes are deposited in GenBank with the accession numbers MN706602–MN706607. Details are as follows: Bacho Kiro AA7-738, MN706602; Bacho Kiro BB7-240, MN706603; Bacho Kiro BK-1653, MN706604; Bacho Kiro CC7-335, MN706605; Bacho Kiro CC7-2289, MN706606; and Bacho Kiro molar F6-620, MN706607.


  1. 1.

    Hublin, J.-J. The modern human colonization of western Eurasia: when and where? Quat. Sci. Rev. 118, 194–210 (2015).

    ADS  Google Scholar 

  2. 2.

    Kuhn, S. L. & Zwyns, N. Rethinking the initial Upper Paleolithic. Quat. Int. 347, 29–38 (2014).

    Google Scholar 

  3. 3.

    Fewlass, H. et al. A 14C chronology for the Middle to Upper Palaeolithic transition at Bacho Kiro cave, Bulgaria. Nat. Ecol. Evol. (2020).

  4. 4.

    White, R. Personal ornaments from the Grotte du Renne at Arcy-sur-Cure. Athena Review 2, 41–46 (2001).

    Google Scholar 

  5. 5.

    Welker, F. et al. Palaeoproteomic evidence identifies archaic hominins associated with the Châtelperronian at the Grotte du Renne. Proc. Natl Acad. Sci. USA 113, 11162–11167 (2016).

    CAS  PubMed  Google Scholar 

  6. 6.

    Hublin, J.-J. et al. Radiocarbon dates from the Grotte du Renne and Saint-Césaire support a Neandertal origin for the Châtelperronian. Proc. Natl Acad. Sci. USA 109, 18743–18748 (2012).

    ADS  CAS  PubMed  Google Scholar 

  7. 7.

    Hublin, J.-J., Spoor, F., Braun, M., Zonneveld, F. & Condemi, S. A late Neanderthal associated with Upper Palaeolithic artefacts. Nature 381, 224–226 (1996).

    ADS  CAS  PubMed  Google Scholar 

  8. 8.

    Ruebens, K., McPherron, S. J. P. & Hublin, J.-J. On the local Mousterian origin of the Châtelperronian: integrating typo-technological, chronostratigraphic and contextual data. J. Hum. Evol. 86, 55–91 (2015).

    PubMed  Google Scholar 

  9. 9.

    Higham, T. et al. The earliest evidence for anatomically modern humans in northwestern Europe. Nature 479, 521–524 (2011).

    ADS  CAS  PubMed  Google Scholar 

  10. 10.

    Benazzi, S. et al. Early dispersal of modern humans in Europe and implications for Neanderthal behaviour. Nature 479, 525–528 (2011).

    ADS  CAS  PubMed  Google Scholar 

  11. 11.

    White, M. & Pettitt, P. Ancient digs and modern myths: the age and context of the Kent’s Cavern 4 maxilla and the earliest Homo sapiens specimens in Europe. Eur. J. Archaeol. 15, 392–420 (2012).

    Google Scholar 

  12. 12.

    Zilhão, J., Banks, W. E., d’Errico, F. & Gioia, P. Analysis of site formation and assemblage integrity does not support attribution of the Uluzzian to modern humans at Grotta del Cavallo. PLoS ONE 10, e0131181 (2015).

    PubMed  PubMed Central  Google Scholar 

  13. 13.

    Kozłowski, J. K. Excavation in the Bacho Kiro Cave (Bulgaria): Final Report. 172 (Panstwowe Wydawnictwo Naukowe, 1982).

  14. 14.

    Hedges, R. E. M., Housley, R. A., Bronk Ramsey, C. & Klinken, G. J. V. Radiocarbon dates from the Oxford AMS system: archaeometry datelist 18. Archaeometry 36, 337–374 (1994).

    Google Scholar 

  15. 15.

    Tsanova, T. & Bordes, J. G. in The Humanized Mineral World: Towards Social and Symbolic Evaluation of Prehistoric Technologies in South Eastern Europe (Proceedings of the ESF Workshop) (eds Tsonev, T. S. & Montagnari Kokclj, E.) 41–50 (ERAUL, 2003).

  16. 16.

    Bailey, S. E. A closer look at Neanderthal postcanine dental morphology: the mandibular dentition. Anat. Rec. 269, 148–156 (2002).

    PubMed  Google Scholar 

  17. 17.

    Bailey, S. E., Skinner, M. M. & Hublin, J.-J. What lies beneath? An evaluation of lower molar trigonid crest patterns based on both dentine and enamel expression. Am. J. Phys. Anthropol. 145, 505–518 (2011).

    PubMed  Google Scholar 

  18. 18.

    Keith, A. Problems relating to the teeth of the earlier forms of prehistoric man. Proc. R. Soc. Med. 6, 103–124 (1913).

    CAS  PubMed  PubMed Central  Google Scholar 

  19. 19.

    Shaw, J. The Teeth, the Bony Palate and the Mandible in Bantu Races of South Africa (Bale and Danielsson, London, 1938).

    Google Scholar 

  20. 20.

    Kallay, J. in Dental Anthropology (ed. Brothwell, D.) 75–86 (Pergamon, 1963).

  21. 21.

    Buckley, M., Collins, M., Thomas-Oates, J. & Wilson, J. C. Species identification by analysis of bone collagen using matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. Rapid Commun. Mass Spectrom. 23, 3843–3854 (2009).

    CAS  PubMed  Google Scholar 

  22. 22.

    Dabney, J. et al. Complete mitochondrial genome sequence of a Middle Pleistocene cave bear reconstructed from ultrashort DNA fragments. Proc. Natl Acad. Sci. USA 110, 15758–15763 (2013).

    ADS  CAS  PubMed  Google Scholar 

  23. 23.

    Korlević, P. et al. Reducing microbial and human contamination in DNA extractions from ancient bones and teeth. Biotechniques 59, 87–93 (2015).

    PubMed  Google Scholar 

  24. 24.

    Gansauge, M.-T. et al. Single-stranded DNA library preparation from highly degraded DNA using T4 DNA ligase. Nucleic Acids Res. 45, e79 (2017).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. 25.

    Fu, Q. et al. DNA analysis of an early modern human from Tianyuan Cave, China. Proc. Natl Acad. Sci. USA 110, 2223–2227 (2013).

    ADS  CAS  PubMed  Google Scholar 

  26. 26.

    Lippold, S. et al. Human paternal and maternal demographic histories: insights from high-resolution Y chromosome and mtDNA sequences. Investig. Genet. 5, 13 (2014).

    PubMed  PubMed Central  Google Scholar 

  27. 27.

    Kivisild, T. Maternal ancestry and population history from whole mitochondrial genomes. Investig. Genet. 6, 3 (2015).

    PubMed  PubMed Central  Google Scholar 

  28. 28.

    Fu, Q. et al. Genome sequence of a 45,000-year-old modern human from western Siberia. Nature 514, 445–449 (2014).

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  29. 29.

    Fu, Q. et al. A revised timescale for human evolution based on ancient mitochondrial genomes. Curr. Biol. 23, 553–559 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  30. 30.

    van der Made, J. in The Encyclopedia of Archaeological Sciences (ed. López Varela, S. L.) 1–4 (Wiley-Blackwell, 2018).

  31. 31.

    Guérin, C. Première biozonation du Pléistocène Européen, principal résultat biostratigraphique de l’étude des Rhinocerotidae (Mammalia, Perissodactyla) du Miocène terminal au Pléistocène supérieur d’Europe Occidentale. Geobios 15, 593–598 (1982).

    Google Scholar 

  32. 32.

    Kuhn, S. L. et al. The early Upper Paleolithic occupations at Uçağizli Cave (Hatay, Turkey). J. Hum. Evol. 56, 87–113 (2009).

    PubMed  Google Scholar 

  33. 33.

    Kuhn, S. L. Upper Paleolithic raw material economies at Üçagizh cave, Turkey. J. Anthropol. Archaeol. 23, 431–448 (2004).

    Google Scholar 

  34. 34.

    Müller, U. C. et al. The role of climate in the spread of modern humans into Europe. Quat. Sci. Rev. 30, 273–279 (2011).

    ADS  Google Scholar 

  35. 35.

    Hershkovitz, I. et al. Levantine cranium from Manot Cave (Israel) foreshadows the first European modern humans. Nature 520, 216–219 (2015).

    ADS  CAS  PubMed  Google Scholar 

  36. 36.

    Fu, Q. et al. An early modern human from Romania with a recent Neanderthal ancestor. Nature 524, 216–219 (2015).

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  37. 37.

    Dibble, H. L. & Lenoir, M. The Middle Paleolithic Site of Combe-Capelle Bas (France) (The University Museum Press, 1995).

  38. 38.

    Turq, A. et al. in Les Sociétés du Paléolithique dans un Grand Sud-ouest de la France: Nouveaux Gisements, Nouveaux Résultats, Nouvelles Méthodes (eds. Jaubert, J. et al.) 83–94 (Mémoire de la Société Préhistorique Française, 2008).

  39. 39.

    Chase, P. G., Debénath, A., Dibble, H. L. & McPherron, S. P. in The Cave of Fontéchevade: Recent Excavations and their Paleoanthropological Implications (eds Chase, P. G. et al.) 28–62 (Cambridge Univ. Press, 2009).

  40. 40.

    Soressi, M. et al. Neandertals made the first specialized bone tools in Europe. Proc. Natl Acad. Sci. USA 110, 14186–14190 (2013).

    ADS  CAS  PubMed  Google Scholar 

  41. 41.

    Richter, D. et al. The age of the hominin fossils from Jebel Irhoud, Morocco, and the origins of the Middle Stone Age. Nature 546, 293–296 (2017).

    ADS  CAS  PubMed  Google Scholar 

  42. 42.

    Sandgathe, D. M., Dibble, H. L., McPherron, S. J. P. & Goldberg, P. in The Middle Paleolithic Site of Pech de l’Azé IV Cave and Karst Systems of the World (eds Dibble, H. L. et al.) 1–19 (Springer, 2018).

  43. 43.

    Sinet-Mathiot, V. et al. Combining ZooMS and zooarchaeology to study Late Pleistocene hominin behaviour at Fumane (Italy). Sci. Rep. 9, 12350 (2019).

    ADS  PubMed  PubMed Central  Google Scholar 

  44. 44.

    Wilson, J., van Doorn, N. L. & Collins, M. J. Assessing the extent of bone degradation using glutamine deamidation in collagen. Anal. Chem. 84, 9041–9048 (2012).

    CAS  PubMed  Google Scholar 

  45. 45.

    Welker, F. et al. Variations in glutamine deamidation for a Châtelperronian bone assemblage as measured by peptide mass fingerprinting of collagen. Sci. Technol. Archaeol. Res. 3, 15–27 (2017).

    Google Scholar 

  46. 46.

    Fewlass, H. et al. Pretreatment and gaseous radiocarbon dating of 40–100 mg archaeological bone. Sci. Rep. 9, 5342 (2019).

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  47. 47.

    Longin, R. New method of collagen extraction for radiocarbon dating. Nature 230, 241–242 (1971).

    ADS  CAS  PubMed  Google Scholar 

  48. 48.

    Brown, T. A., Nelson, D. E., Vogel, J. S. & Southon, J. R. Improved collagen extraction by modified Longin method. Radiocarbon 30, 171–177 (1988).

    CAS  Google Scholar 

  49. 49.

    Bronk Ramsey, C., Higham, T., Bowles, A. & Hedges, R. Improvements to the pretreatment of bone at Oxford. Radiocarbon 46, 155–163 (2004).

    Google Scholar 

  50. 50.

    Brock, F., Bronk Ramsey, C. & Higham, T. Quality assurance of ultrafiltered bone dating. Radiocarbon 49, 187–192 (2007).

    CAS  Google Scholar 

  51. 51.

    Wacker, L., Němec, M. & Bourquin, J. A revolutionary graphitisation system: fully automated, compact and simple. Nucl. Instrum. Methods Phys. Res. B 268, 931–934 (2010).

    ADS  CAS  Google Scholar 

  52. 52.

    Wacker, L. et al. MICADAS: routine and high-precision radiocarbon dating. Radiocarbon 52, 252–262 (2010).

    CAS  Google Scholar 

  53. 53.

    Korlević, P., Talamo, S. & Meyer, M. A combined method for DNA analysis and radiocarbon dating from a single sample. Sci. Rep. 8, 4127 (2018).

    ADS  PubMed  PubMed Central  Google Scholar 

  54. 54.

    Wacker, L., Christl, M. & Synal, H. A. Bats: a new tool for AMS data reduction. Nucl. Instrum. Methods Phys. Res. B 268, 976–979 (2010).

    ADS  CAS  Google Scholar 

  55. 55.

    Reimer, P. J. et al. IntCal13 and Marine13 radiocarbon age calibration curves 0–50,000 years cal BP. Radiocarbon 55, 1869–1887 (2013).

    CAS  Google Scholar 

  56. 56.

    Bronk Ramsey, C. Bayesian analysis of radiocarbon dates. Radiocarbon 51, 337–360 (2009).

    Google Scholar 

  57. 57.

    Skinner, M. M., Gunz, P., Wood, B. A. & Hublin, J.-J. Enamel–dentine junction (EDJ) morphology distinguishes the lower molars of Australopithecus africanus and Paranthropus robustus. J. Hum. Evol. 55, 979–988 (2008).

    PubMed  Google Scholar 

  58. 58.

    Skinner, M. M., Gunz, P., Wood, B. A., Boesch, C. & Hublin, J.-J. Discrimination of extant Pan species and subspecies using the enamel–dentine junction morphology of lower molars. Am. J. Phys. Anthropol. 140, 234–243 (2009).

    Google Scholar 

  59. 59.

    Slon, V. et al. Neandertal and Denisovan DNA from Pleistocene sediments. Science 356, 605–608 (2017).

    ADS  CAS  PubMed  Google Scholar 

  60. 60.

    Glocke, I. & Meyer, M. Extending the spectrum of DNA sequences retrieved from ancient bones and teeth. Genome Res. 27, 1230–1237 (2017).

    CAS  PubMed  PubMed Central  Google Scholar 

  61. 61.

    Dabney, J. & Meyer, M. Length and GC-biases during sequencing library amplification: a comparison of various polymerase-buffer systems with ancient and modern DNA sequencing libraries. Biotechniques 52, 87–94 (2012).

    CAS  Google Scholar 

  62. 62.

    Kircher, M., Sawyer, S. & Meyer, M. Double indexing overcomes inaccuracies in multiplex sequencing on the Illumina platform. Nucleic Acids Res. 40, e3 (2012).

    CAS  PubMed  Google Scholar 

  63. 63.

    Renaud, G., Stenzel, U. & Kelso, J. leeHom: adaptor trimming and merging for Illumina sequencing reads. Nucleic Acids Res. 42, e141 (2014).

    PubMed  PubMed Central  Google Scholar 

  64. 64.

    Li, H. & Durbin, R. Fast and accurate long-read alignment with Burrows–Wheeler transform. Bioinformatics 26, 589–595 (2010).

    PubMed  PubMed Central  Google Scholar 

  65. 65.

    Meyer, M. et al. A high-coverage genome sequence from an archaic Denisovan individual. Science 338, 222–226 (2012).

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  66. 66.

    Li, H. et al. The Sequence Alignment/Map format and SAMtools. Bioinformatics 25, 2078–2079 (2009).

    PubMed  PubMed Central  Google Scholar 

  67. 67.

    Meyer, M. et al. A mitochondrial genome sequence of a hominin from Sima de los Huesos. Nature 505, 403–406 (2014).

    ADS  CAS  PubMed  Google Scholar 

  68. 68.

    Meyer, M. et al. Nuclear DNA sequences from the Middle Pleistocene Sima de los Huesos hominins. Nature 531, 504–507 (2016).

    ADS  CAS  PubMed  Google Scholar 

  69. 69.

    Green, R. E. et al. A complete Neandertal mitochondrial genome sequence determined by high-throughput sequencing. Cell 134, 416–426 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  70. 70.

    Benazzi, S. et al. The makers of the Protoaurignacian and implications for Neandertal extinction. Science 348, 793–796 (2015).

    ADS  CAS  PubMed  Google Scholar 

  71. 71.

    Ermini, L. et al. Complete mitochondrial genome sequence of the Tyrolean Iceman. Curr. Biol. 18, 1687–1693 (2008).

    CAS  PubMed  Google Scholar 

  72. 72.

    Gilbert, M. T. P. et al. Paleo-Eskimo mtDNA genome reveals matrilineal discontinuity in Greenland. Science 320, 1787–1789 (2008).

    ADS  CAS  PubMed  Google Scholar 

  73. 73.

    Krause, J. et al. A complete mtDNA genome of an early modern human from Kostenki, Russia. Curr. Biol. 20, 231–236 (2010).

    CAS  PubMed  Google Scholar 

  74. 74.

    Green, R. E. et al. A draft sequence of the Neandertal genome. Science 328, 710–722 (2010).

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  75. 75.

    Posth, C. et al. Deeply divergent archaic mitochondrial genome provides lower time boundary for African gene flow into Neanderthals. Nat. Commun. 8, 16046 (2017).

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  76. 76.

    Prüfer, K. et al. The complete genome sequence of a Neanderthal from the Altai Mountains. Nature 505, 43–49 (2014).

    ADS  Google Scholar 

  77. 77.

    Rougier, H. et al. Neandertal cannibalism and Neandertal bones used as tools in Northern Europe. Sci. Rep. 6, 29005 (2016).

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  78. 78.

    Skoglund, P. et al. Separating endogenous ancient DNA from modern day contamination in a Siberian Neandertal. Proc. Natl Acad. Sci. USA 111, 2229–2234 (2014).

    ADS  CAS  PubMed  Google Scholar 

  79. 79.

    Krause, J. et al. The complete mitochondrial DNA genome of an unknown hominin from southern Siberia. Nature 464, 894–897 (2010).

    ADS  CAS  PubMed  Google Scholar 

  80. 80.

    Reich, D. et al. Genetic history of an archaic hominin group from Denisova Cave in Siberia. Nature 468, 1053–1060 (2010).

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  81. 81.

    Sawyer, S. et al. Nuclear and mitochondrial DNA sequences from two Denisovan individuals. Proc. Natl Acad. Sci. USA 112, 15696–15700 (2015).

    ADS  CAS  PubMed  Google Scholar 

  82. 82.

    Slon, V. et al. A fourth Denisovan individual. Sci. Adv. 3, e1700186 (2017).

    ADS  PubMed  PubMed Central  Google Scholar 

  83. 83.

    Horai, S. et al. Man’s place in Hominoidea revealed by mitochondrial DNA genealogy. J. Mol. Evol. 35, 32–43 (1992).

    ADS  CAS  PubMed  Google Scholar 

  84. 84.

    Katoh, K. & Standley, D. M. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol. Biol. Evol. 30, 772–780 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  85. 85.

    Kumar, S., Stecher, G. & Tamura, K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol. Biol. Evol. 33, 1870–1874 (2016).

    CAS  Google Scholar 

  86. 86.

    Schliep, K. P. phangorn: phylogenetic analysis in R. Bioinformatics 27, 592–593 (2011).

    CAS  PubMed  Google Scholar 

  87. 87.

    Kloss-Brandstätter, A. et al. HaploGrep: a fast and reliable algorithm for automatic classification of mitochondrial DNA haplogroups. Hum. Mutat. 32, 25–32 (2011).

    PubMed  Google Scholar 

  88. 88.

    Renaud, G., Slon, V., Duggan, A. T. & Kelso, J. Schmutzi: estimation of contamination and endogenous mitochondrial consensus calling for ancient DNA. Genome Biol. 16, 224 (2015).

    PubMed  PubMed Central  Google Scholar 

  89. 89.

    Bouckaert, R. et al. BEAST 2: a software platform for Bayesian evolutionary analysis. PLOS Comput. Biol. 10, e1003537 (2014).

    PubMed  PubMed Central  Google Scholar 

  90. 90.

    Darriba, D., Taboada, G. L., Doallo, R. & Posada, D. jModelTest 2: more models, new heuristics and parallel computing. Nat. Methods 9, 772 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  91. 91.

    Baele, G. et al. Improving the accuracy of demographic and molecular clock model comparison while accommodating phylogenetic uncertainty. Mol. Biol. Evol. 29, 2157–2167 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  92. 92.

    Stoops, G. Guidelines for Analysis and Description of Soil and Regolith Thin Sections. 184 (Soil Science Society of America, 2003).

  93. 93.

    Courty, M. A., Goldberg, P. & Macphail, R. Soils and Micromorphology in Archaeology 344 (Cambridge Univ. Press, 1989).

Download references


We thank the tourism association of Bacho Kiro Cave in the town of Dryanovo, the History museum – Dryanovo, the Regional History museum in the city of Gabrovo, Dryanovo town hall and V. Lafchiiski for their assistance with the fieldwork and in the laboratory; N. Spassov from the National Museum of Natural History in Sofia for cooperating and hosting researchers of our project; H. Temming and J. Honeyford for their technical assistance and S. Nagel, B. Nickel, B. Schellbach and A. Weihmann for their help with the ancient DNA laboratory procedures and sequencing. Field operations were funded by the Max Planck Society. AixMICADAS and its operation are funded by Collège de France and the EQUIPEX ASTER-CEREGE (principal investigator, E.B.). S.T. is funded by the European Research Council under the European Union’s Horizon 2020 Research and Innovation Programme (grant agreement no. 803147-951 RESOLUTION). The ancient DNA part of this study was funded by the Max Planck Society and the European Research Council (grant agreement no. 694707 to S.P.).

Author information




J.-J.H. designed the study. T. Tsanova, N.S., V.A., S.S., R.S., E.E., Z.R. and S.P.M. collected field data; H.F., B.K., L.W., E.B., Y.F., T. Tuna and S.T. established the radiocarbon dates; V.A. studied the micromorphology of the sediments; S.B., M.M.S. and J.-J.H. analysed hominin dental morphology; V.S.-M., L.P., F.W. and A.W. performed ZooMS; M.H., M.M. and S.P. performed mtDNA analysis; T. Tsanova, N.S., N.Z., S.S., I.K., V.D., J.M. and S.P.M. conducted the study of the lithics; G.M.S., R.S., V.P. and N.L.M. analysed the faunal assemblages and the osseous objects. J.-J.H. wrote the paper with contributions of all authors.

Corresponding author

Correspondence to Jean-Jacques Hublin.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Peer review information Nature thanks William Banks, Richard G. Klein and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data figures and tables

Extended Data Fig. 1 Excavations at Bacho Kiro Cave, 2015–2018.

a, Plan view of the entrance and the excavated areas of the cave, with the grid system of our recent excavations (letters in the left column) and those of the 1971–1975 excavations (letters in the right column). b, Site location in southeastern Europe. c, Photograph of the entrance of the cave. The floor is artificially raised; the original entrance was several metres lower than shown in this photograph. d, Initial stratigraphic section drawing of the exposed profile from the Main sector in 2015 (codes for the archaeological layers are on the left, with the corresponding layers from the 1971–1975 excavations in parentheses). e, Frontal view of the Niche 1 sector and its stratigraphic subdivisions. f, Lower part of the stratigraphic section drawing of the Niche 1 sector, in 2018. Note the thickness and preservation of the lower deposits here in comparison with the Main sector profile. g, Photograph of the Main sector transversal section on the line between squares F5–F6 and squares G5–G6 before excavation in 2015. CF, combustion feature. hn, Hominin remains identified by ZooMS with their IDs: BK-1653 (h) and F6-597 (j) from layer B, with h coming from the 1971–1975 excavations (dashed line); BB7-240 (k), CC7-2289 (l), CC7-335 (m) and AA7-738 (n) from layer N1-I. Continuous lines connect the fossils with their find locations. i, Second lower molar (F6-620) from layer J in the Main sector.

Extended Data Fig. 2 Geographical distributions.

Geographical distribution of the main IUP sites of western and central Eurasia (black dots), directly dated early H. sapiens predating 37,000 cal. bp (empty black dots) and directly dated late Neanderthals associated with Châtelperronian assemblages (orange squares). Bacho Kiro Cave is represented by a red circle.

Extended Data Fig. 3 Photographs of lithic artefacts from layer I of Bacho Kiro Cave.

Pointed retouched blades and fragments (1–4, 6, 7) and piece with bifacial retouch (5). Photographs by V.S.-M. and T. Tsanova.

Extended Data Fig. 4 Drawings of lithic artefacts from layer I of Bacho Kiro Cave.

Pointed retouched blade with slightly oblique truncation and base modified by inverse retouch (1), pointed blade fragments (2 and 5, which has an oblique truncation and slight notch on the left edge, and was perhaps intentionally fragmented), pointed, small blades fragments (3, 7, 8 and 9), pointed blade fragment with opposing pseudo-burin blows on the apex and on the distal fracture edge (perhaps indicating use as a projectile) (4) and Levallois flake (6). Drawings by I.K. and T. Tsanova).

Extended Data Fig. 5 Human lower second molar (F6-620).

a, Mesial, buccal and distal views of the crown, root and pulp chamber (left) and occlusal views of the enamel and dentine crown (right). b, A principal component analysis of the shape of the enamel–dentine junction ridge and cervix places the Bacho Kiro Cave second lower molar (F6-620) represented by a red star within the samples of recent (n = 8) and Pleistocene (n = 9) H. sapiens, and outside the distribution of Neanderthals (n = 20) and H. erectus (n = 3).

Extended Data Fig. 6 MALDI–TOF MS spectra for the six bone specimens identified as hominins through ZooMS analysis.

a, B4-1653 (interface of layers 6a and 7). b, AA7-738 (layer N1-I). c, BB7-240 (layer N1-I). d, CC7-2289 (layer N1-I). e, CC7-335 (layer N1-I). f, F6-597 (layer B).

Extended Data Fig. 7 Frequency of nucleotide substitutions at the beginning and the ends of mtDNA alignments for the Bacho Kiro Cave specimens.

Only fragments of at least 35 base pairs in length that mapped to the revised Cambridge Reference Sequence with a mapping quality of at least 25 were used for this analysis. Solid lines in red depict all fragments and dashed lines depict the fragments that have a C-to-T substitution at the opposing end (‘conditional’ C-to-T substitutions). All other types of substitution are marked in grey.

Extended Data Fig. 8 Bayesian phylogenetic tree relating Bacho Kiro Cave mtDNA to 54 present-day humans, 10 directly radiocarbon dated ancient H. sapiens and the Vindija 33.16 Neanderthal.

The Bacho Kiro Cave specimens are in red. Other ancient H. sapiens used as calibration points to estimate the tip dates of Bacho Kiro Cave specimens are italicized. The posterior probabilities are denoted above the branches. The mtDNA of Vindija 33.16 was used to root the tree (not shown).

Extended Data Table 1 Comparative dental metrics
Extended Data Table 2 mtDNA branch-shortening estimates

Supplementary information

Supplementary Information

This file contains Supplementary Discussion sections 1-7 with Supplementary Figures 1-10, Supplementary Tables 1-16 and additional references.

Reporting Summary

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Hublin, J., Sirakov, N., Aldeias, V. et al. Initial Upper Palaeolithic Homo sapiens from Bacho Kiro Cave, Bulgaria. Nature 581, 299–302 (2020).

Download citation

Further reading


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.