The aim of this study was to investigate seasonal variations in the prevalence of the nasopharyngeal carriage of respiratory pathogens and identify factors affecting colonisation patterns in healthy children. The nasopharyngeal carriage of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis during two seasons (autumn and spring) was evaluated in 1580 healthy children aged 1-7 years by means of a cohort study conducted in day-care centres and schools in eight Italian cities. A questionnaire was used to obtain the epidemiological data. In all, 309 children (19.5%) carried one or more respiratory pathogens in the autumn, and 375 children (23.7%) in the spring. This variation was due to H. influenzae alone or in combination (autumn: S. pneumoniae 60, 3.8%; H. influenzae 206, 13.0%, M. catarrhalis 71, 4.5%; spring: S. pneumoniae 75, 4.7%; H. influenzae 288, 18.2%, M. catarrhalis 82, 5.2%). Colonisation with two or more pathogens increased from 9.1% in the spring to 17.3% in the autumn. Seasonal variations occur in the prevalence of the nasopharyngeal carriage of respiratory pathogens in healthy children attending day-care centres or schools in Italy. However, although statistically significant, the difference was slight and had limited clinical relevance. Therefore, seasonal influence on the nasopharyngeal carriage of respiratory pathogens in healthy children was negligible.