Molecular identification of pathogens in ancient skeletal remains from Butrint and Diaporit, Albania

Paleopathologists and anthropologists examine skeletal lesions to better understand diseases that afflicted ancient societies. Traditional analyses of skeletal remains have limited success in identifying the particular organism or pathogen responsible. These limitations can be overcome by the addition of molecular techniques to screen for pathogen DNA present in the remains. Analysis of skeletal material from a World Heritage Site in Butrint and Diaporit, Albania demonstrated that pathogen DNA analysis could be a useful tool to the study of ancient cultures. Bone samples were collected from five sets of remains that showed pathologies, three late Roman (5th--7 th century AD) and two from the late Medieval period (11th --13th century AD). Skeletal lesions seen in these remains had commonality indicating that the same pathogen may have been involved. In particular, lesions present on the vertebrae and ribs of the late Medieval samples were almost identical. Bone DNA extracts were screened for the presence of DNA from the causative agents of tuberculosis and brucellosis. Real-time PCR results indicated that the vertebrae and ribs of the late Medieval samples contained Brucella spp. DNA. This provides novel information on the ancient inhabitants of a culturally rich site that would have otherwise been unobtainable. This also represents the first time brucellosis has been molecularly identified in an ancient society and proved that molecular techniques can be used to investigate diseases processes in other skeletal populations and cultures.