Neolithic and medieval virus genomes reveal complex evolution of hepatitis B

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