A cross-sectional epidemiological study has been undertaken to relate the bacterial composition of approximal dental plaque with the earliest stages of caries development in schoolchildren. Small samples of plaque were removed from multiple sites around the contact areas of 42 premolars extracted for orthodontic reasons from 29 schoolchildren (mean age = 13.5 yr). Caries diagnosis was based on polarized light microscopy and contact microradiography of thin sections cut through the sample sites. Fifty-seven percent of sites (37l60) showed histological evidence of demineralization. Both the isolation frequency and the mean percentage viable count of mutans streptococci and Actinomyces viscosus were higher at sites with early caries, although mutans streptococci could not be detected at 37% of sites with early caries. At these latter sites, the proportions of Veillonella were markedly reduced. Lactobacilli were rarely isolated and were never recovered from caries-free surfaces. Analysis of the data shows that the relationship between plaque bacteria and enamel is neither merely passive nor indifferent, and that particular stages of lesion formation may be associated with different combinations of bacteria.