In vitro Studies of Growth and Competition between S. salivarius TOVE-R and Mutans Streptococci

Abstract
Streptococcus salivarius TOVE-R has previously been reported as a successful competitor with Streptococcus mutans 10449S and Streptococcus sobrinus 6715-13 WT on the teeth of rats. We studied, in vitro, some possible bases for this competition, including hydrogen peroxide or catalase production, bacteriocin or enocin production, and growth rates. Growth rates were measured spectrophotometrically in both complex and defined media. We studied conditions of (i) aerobic and anaerobic incubation; (ii) glucose and sucrose medium supplementation at three concentrations each; (iii) various initial pH's; (iv) singly- or doubly-inoculated cultures; and (v) half-strength, normal, or double-strength broth used both fresh and as culture filtrates. TOVE-R grew as well as did mutans streptococci at acid pH, decidedly better at alkaline pH, and nearly twice as fast near neutral pH. The average doubling time for TOVE-R was about 0.5 hr, while that for the mutans streptococci was about 1.0 hr. When TOVE-R was grown together with a mutans streptococcus, the growth rate observed for the doubly-inoculated culture was equal to or less than that of TOVE-R alone, never greater. The presence and proportions of both organisms in mixed cultures were confirmed by plate counts, direct specific immunofluorescence, and Nomarski interference microscopy. There was no evidence, by any of the methods employed, to indicate the production of an inhibitory substance against mutans streptococci by TOVE-R, or vice versa. Also, there was no evidence that the faster growth rate of TOVE-R could be attributed to nutrient limitation of the mutans streptococci. TOVE-R's ability to compete successfully with mutans streptococci in vivo may be attributable to its inherent ability to grow faster over a wide range of environmental conditions, as observed in vitro.