Bacteriological effects of a Lactobacillus reuteri probiotic on in vitro oral biofilms

Despite the targeted and incidental exposure of the oral cavity to probiotic bacteria, relatively little information is available concerning their bacteriological effects or ecological fate in this environment. We have investigated the effects of an oral probiotic bacterium, Lactobacillus reuteri on salivary biofilm microcosms. Nascent in vitro plaques in hydroxyapatite disc model and mature continuous culture plaques within constant depth film fermenters (CDFFs) were exposed to probiotic strains of L. reuteri. Microbial composition was assessed using differential viable counting and CDFF plaques were additionally characterised using qPCR and PCR-DGGE. Probiotic dosing of nascent plaques was associated with significant increases in lactobacilli and Gram-negative anaerobes in both biofilm and planktonic phases, and with elevated counts of streptococci in the planktonic phase that were not accompanied by decreases in pH. In mature CDFF plaques, differential culture indicated that whilst significant increases in lactobacillus counts occurred during dosing, numbers of other bacterial groups and pH did not significantly change. Monitoring the probiotic strain in CDFFs using selective culture and qPCR indicated that the exogenous bacterium reached and maintained c. 5 log10 CFU/mm2 during dosing and 20 days after cessation. The apparent lack of marked bacterial inhibition by L. reuteri in both model systems was supported by the absence of antagonism in binary antagonism assays. The largest compositional changes occurred in nascent plaques where the addition of lactobacilli caused significant increases in streptococci and Gram-negative anaerobes, the latter possibly via lactate syntrophy. Exogenous L. reuteri persisted in mature plaque microcosms after dosing ceased.