34The hepatitis B virus (HBV) is one of the most widespread human pathogens known today, 35 yet its origin and evolutionary history are still unclear and controversial. Here, we report the 36 analysis of three ancient HBV genomes recovered from human skeletons found at three 37 different archaeological sites in Germany. We reconstructed two Neolithic and one medieval 38 HBV genomes by de novo assembly from shotgun DNA sequencing data. Additionally, we 39 observed HBV-specific peptides using paleo-proteomics. Our results show that HBV 40 circulates in the European population for at least 7000 years. The Neolithic HBV genomes 41show a high genomic similarity to each other. In a phylogenetic network, they do not group 42 with any human-associated HBV genome and are most closely related to those infecting 43African non-human primates. These ancient virus forms appear to represent distinct lineages 44 that have no close relatives today and went possibly extinct. Our results reveal the great 45 potential of ancient DNA from human skeletons in order to study the long-time evolution of 46 blood borne viruses. 47 48 49

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