Supplementary Material for: Association of Root Biofilm Bacteriome with Root Caries Lesion Severity and Activity

Introduction: This research aimed to assess the association of root biofilm bacteriome with root caries lesion severity and activity in institutionalised Colombian elderlies and was conducted to gather data on the root caries bacteriome in this population. Methods: A bacteriome evaluation of biofilm samples from sound, and carious root surfaces was performed. Root caries was categorised (ICDAS Root criteria) based on severity (Sound surfaces, Initial: non-cavitated, Moderate/Extensive combined: cavitated) and activity status (Active and Inactive). DNA was extracted and the V4 region of the 16S rRNA gene was sequenced, after the classification of features employing amplicon sequence variants (ASVs) and taxonomic assignment via the Human Oral Microbiome Database (HOMD). Bacterial richness, diversity (Simpson's and Shannon's indices), and relative abundance estimation were assessed and compared based on root caries severity and activity status (including Sound surfaces). Results: A total of 130 biofilm samples were examined: Sound (n=45) and with root caries lesions (n=85; Initial: n=41; Moderate/Extensive: n=44; Active: n=60; Inactive: n=25). Species richness was significantly lower in biofilms from Moderate/Extensive and Active groups compared to Sound sites. There was a higher relative abundance of species like Lechtotricia wadei, Capnocytophaga granulosa, Cardiobacterium valvarum, Porphyromonas pasteri -in Sound sites; Dialister invisus, Streptococcus mutans, Pseudoramibacter alactolyticus and Bacteroidetes (G-5) bacterium 511 -in Moderate/Extensive lesions, and Fusobacterium nucleatum subsp. animalis, Prevotella denticola, Lactobacillus fermentum, Saccharibacteria (TM7) [G-5] bacterium HMT 356 -in active lesions. Conclusion: Root caries bacteriome exhibited differences in species proportions between the compared groups. Specifically, cavitated lesions and active lesions showed higher relative abundance of acidogenic bacteria.