It was the purpose of the present investigation to monitor the composition of the subgingival microbiota at selected sites in individuals passing through puberty and to correlate observed changes with the development of pubertal maturation. Between the ages of 11 and 14 years, pubertal and skeletal maturation was monitored annually in 22 boys and 20 girls. During this time, subgingival microbial samples were taken every 4th to 5th month (10 times in 4 years) mesially of the upper first molars. High values in total bacterial counts were reached after the onset of puberty, followed by a decrease towards the end of the observation period. The frequency of detection of Actinomyces odontolyticus and of Capnocytophaga sp. increased with time. The frequencies of other selected species, specifically of black pigmenting Bacteriodes sp. were not found to increase when tested by linear and quadratic models of time trend. However, a statistically significant rise in the frequency of detecting B. intermedius and B. melaninogenicus was noted in the initial pubertal phase identified by the onset of testicular growth in boys (p = 0.05). A significant relationship also existed between testes growth and increase of A. odontolyticus (p less than 0.01). In girls, a similar increase was obtained for A. odontolyticus when studied in relation to the Tanner scores for breast development (p less than 0.01). The changes observed in the subgingival microbiota during puberty may be related to the development of gingivitis, which was demonstrated by a higher tendency for gingival bleeding during the course of the pubertal maturation process.