Title: Tuberculosis in mummies – New findings, perspectives and limitations

Authors: Albert Zink¹, Heidi Y. Jäger¹, Guido Valverde¹, Ildikó Pap², Ildikó Szikossy², György Pálfi², Frank Maixner¹

Affiliations: ¹Institute for Mummy Studies, Eurac Research, Viale Druso 1, 39100 Bolzano, Italy; ²Department of Biological Anthropology, Faculty of Science and Informatics, University of Szeged, Szeged, Hungary

Abstract: The molecular analysis of ancient pathogen DNA represents a unique opportunity for the study of infectious diseases in ancient human remains. Among other diseases, paleogenetic studies have been successful in proving tuberculous DNA in ancient human remains. In the beginning of ancient DNA studies, the presence of TB DNA was assessed using a PCR-based assay targeting specific regions of the MTB complex, such as the repetitive element IS6110. The advent of high-throughput sequencing has enabled the reconstruction of full ancient TB genomes in the field of paleomicrobiology. However, despite the numerous paleopathological and PCR-based studies on the presence of tuberculosis in ancient human remains, full genome wide reconstructions are still limited to well-preserved specimens with low environmental contamination and connected with extensive screening efforts. This has led to some controversies regarding the evolutionary history of its causative agent Mycobacterium tuberculosis. In this context, mummies have been shown to be a good source for the detection of MTB complex DNA due to a low exposure to environmental influences and the overall good state of preservation of hard and soft tissues in the human remains. Here, we present new findings on the presence of mixed TB infections in the 18^th century naturally mummified human remains from Vác, Hungary and the presence of MTB complex DNA in a naturally mummified individual form Pre-Columbian Bolivia. The future perspectives of detecting tuberculosis in mummies will be discussed in the light of methodological aspects, as well as ethical and conservatory challenges.