Evaluation of bacterial nitrate reduction in the human oral cavity

It is hypothesized that the enterosalivary nitrate circulation encourages nitrate reducing bacteria to reside within the oral cavity. Nitrite production may then limit the growth of acidogenic bacteria as a result of the production of antimicrobial oxides of nitrogen, including nitric oxide. This study was carried out with 10 subjects to characterize oral nitrate reduction and identify the bacteria responsible. Nitrate reduction varied between individuals (mean 85.4 +/- 15.9 nmol nitrite min(-1) with 10 ml 1 mm KNO(3) mouth wash) and was found to be concentrated at the rear of the tongue dorsal surface. Nitrate reductase positive isolates identified, using 16S rDNA sequencing, from the tongue comprised Veillonella atypica (34%), Veillonella dispar (24%), Actinomyces odontolyticus (21%), Actinomyces naeslundii (2%), Rothia mucilaginosa (10%), Rothia dentocariosa (3%) and Staphylococcus epidermidis (5%). Nitrite production rates, using intact and permeabilized cells, of the major tongue nitrate reducers were determined in the presence of methyl and benzyl viologen. Under anaerobic conditions in the presence of nitrate, rates in decreasing order were: A. odontolyticus > R. mucilaginosa > R. dentocariosa > V. dispar > V. atypica. In conclusion, Veillonella spp. were found to be the most prevalent taxa isolated and thus may make a major contribution to nitrate reduction in the oral cavity.