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Streptococcus mutans is recognized as the main cause of dental caries, and the formation of a plaque biofilm is required for caries development. This study aimed to determine the inhibitory effect of Lactobacillus plantarum FB-T9 on S. mutans biofilm formation in vitro and on the prevention and treatment of dental caries in rats. During in vitro experiments, FB-T9 exhibited good bacteriostatic ability in a plate competition assay. This strain also significantly reduced the biomass and viability of S. mutans biofilms and induced structural damage during the early (6 h), middle (12 h) and late (24 h) stages of biofilm formation. In a 70-day in vivo experiment, FB-T9 significantly reduced the levels of S. mutans on the dental surfaces of rats by more than 2 orders of magnitude of the levels in the dental caries model group (p < 0.05). Moreover, FB-T9 significantly reduced the caries scores (modified Keyes scoring method) in both the prevention and treatment groups (p < 0.05) and had great colonization potential in the oral cavity. These results indicate the potential usefulness of L. plantarum FB-T9 as a probiotic for the prevention and treatment of caries. [ABSTRACT FROM AUTHOR]