Two hundred and seventeen approximal spaces, initially caries free, in 58 patients were studied clinically and radiographically, at intervals of 6 months, for 3 years. Samples of approximal dental plaque were removed for culture of Streptococcus mutans, Streptococcus sobrinus, and lactobacilli. During the study approximal caries developed in 16 subjects (27.6%) at 30 sites (13.8%), involving 42 teeth. 56 of 58 (96.7%) subjects harboured S. mutans at some time during the study, 62.1% lactobacilli, and 29.3% had S. sobrinus. The counts of S. mutans were significantly higher in those subjects that also carried S. sobrinus than in the remaining subjects. A persisting high count of S. mutans > 10(5) colony-forming units per millilitre (CFU/ml) or a count that rose by > or = 1 x log10 CFU/ml during the study was seen in 25 of 30 sites that subsequently became carious (p 10(3) CFU/ml or rising by > or = 1 x log10 CFU/ml were seen in 17 of 30 sites that became carious (p