Intrapopulation and interpopulation variation in blood pressure (BP) often reflects the joint effect of a complex set of risk factors, including lifestyle factors such as physical activity, diet, smoking and alcohol use. In this study, we set out to quantify the impact of habitual levels of physical activity on BP within and between three populations at contrasting levels of population risk of hypertension. Individuals were randomly sampled from communities in Nigeria (n=57), Jamaica (from Kingston, n=35) and the United States (from the Chicago area, n=32). Activity energy expenditure (AEE) (estimated from resting energy expenditure measured by indirect calorimetry and total expenditure measured with doubly labelled water) was used as an objective estimate of physical activity. In each of the three samples, there was a consistent negative correlation between BP and AEE. This negative association persisted after adjustment for age, sex and body fat (body mass index or percent fat mass). In multivariate models, adiposity was no longer a significant predictor of BP after accounting for low AEE. In conclusion these data suggest that habitual levels of physical activity may have a generalizable relationship with BP in populations with widely different social and environmental characteristics.