The aims of this study were to assess levels and patterns of physical activity (PA) in relation to age and regular sport activity, and to examine its relationship to physical fitness in trained and untrained boys. One hundred forty-seven 11-to 15-year-old boys (73 trained and 74 untrained) participated in this study. Trained boys, comprised of 26 soccer, 25 handball and 22 volleyball players, had been training regularly for at least one year. The intensity, duration and frequency of PA were assessed from four complete days of heart rate monitoring with 15-seconds sampling intervals. Aerobic fitness was assessed by determining peakVO(2) with a portable breath-by-breath gas analyzer (Cosmed K4b(2)) and the running speeds at fixed lactate concentrations during an incremental running test. Anaerobic fitness was evaluated with the Wingate Anaerobic Test. Skinfold thicknesses from eight sites and Tanner stages of pubic hair were also obtained. Based on 15-s heart rate data, instead of continuous activity, multiple short bouts of moderate and vigorous PA, lasting up to one minute, were characteristic of daily PA patterns of both trained and untrained boys. PA levels of trained boys were higher than untrained boys (p < 0.01) and the levels of PA decreased with age and maturation in both groups (p < 0.05). Daily PA variables were related to body fatness in both groups (p < 0.05), but the relationships were not consistent in the trained group. Daily PA variables were also related to aerobic fitness in the untrained group (p < 0.05) and these relationships were somewhat better with vigorous PA, whereas in the trained group, none of the PA variables were related to any of the aerobic fitness indices (p > 0.05). No relationship was observed between PA variables and anaerobic fitness in either group (p> 0.05). It seems that such relationships may somewhat depend on the fitness level of the subjects.