Fusobacterium Nucleatum Dna In Dental Calculus From Ancient Siberian Populations

To assess the possibility of DNA preservation of periodontopathogenic microorganisms in the archaeological material of dental calculus of the Western Siberia populations. A DNA search of periodontopathogenic bacteria was conducted in dental calculus of 38 individuals of indigenous populations in the north of Western Siberia XII-XX centuries. Teeth were washed with a sterile TE buffer and the dental calculus was removed and shredded in the disperser under aseptic conditions. Then DNA extraction was carried out using a fluorochloroplastic method. To check the quality of the extracted DNA, an amplification of fragments of the gene 16S rDNA was conducted using primers to the V4 domain of F515 gene (5′-CACGGTCGKCGGCGCCATT-3′) and R806 gene (5′-GGACTACHVGGGTWTCTAAT-3′). The search for DNA of periodontopathogens was conducted in accordance with the method proposed by [De La Fuente, C., Flores, S., & Moraga, M. DNA from human ancient bacteria: a novel source of genetic evidence from archaeological dental calculus. Archaeometry, 2013. 55(4), 767-778.]. The experiments were repeated three times for each sample. 9 out of 38 cases were positive for DNA amplification of the fadA Fusobacterium nucleatum gene. The obtained sequences showed a high (97.04%-99.4%) percentage of similarity to the sequence of the corresponding gene of Fusobacterium nucleatum subsp. polymorphum strain ChDC F306 (GenBank Acc. # CP013121.1). The evidence of DNA preservation of microorganisms representing the oral cavity microbiota has been obtained, including the periodontopathogenic species Fusobacterium nucleatum in dental calculus from teeth dating to XII-XVIII centuries.