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Besides environmental and psycho-social factors explaining the variation in physical activity levels during adolescence, some evidence suggests that biological processes are involved in regulating habitual daily physical activity and energy expenditure. The purpose of this study was to examine the influence of biological maturity status on physical activity. Chronological age, standing height, sitting height, and body mass were measured cross-sectionally in 268 girls, aged 9.5 to 11.5 years. Biological maturity groups (Early, Average, Late) were created according to estimated age at peak-height-velocity (estAPHV). Habitual physical activity was determined with a pedometer (Yamax Digiwalker SW-200) over a 7-day period. Differences in steps/day across maturity groups were examined by ANCOVA, controlling separately for time the pedometer was worn, leg length, and body mass. Mean pedometer steps/day was 10,822 +/- 2,639. As expected, body size varied by maturity status (e.g., early > average > late). Significant maturity group differences were found with early maturing girls showing lower activity levels compared to average or late maturers. These differences remained after controlling for time the pedometer was worn and leg length; however, the differences were no longer significant when controlling for body mass. The results suggest that biological maturity status influences physical activity levels in girls between 10 and 12 years of age but the relationship is not independent of body mass. Further research is needed to establish the complex inter-relationships among adiposity, biological maturation, and energy expenditure during puberty., (2009 Wiley-Liss, Inc.)