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Evidence supports the health benefits of physical activity, but the longitudinal patterns of physical activity associated with long-term health outcomes remain insufficiently understood. In 231,488 health professionals from three US cohorts with repeated physical activity assessments for 32 years, we prospectively examined the associations of long-term physical activity patterns (volume, consistency, and trajectories) with incidence of major chronic diseases, including type 2 diabetes, major cardiovascular disease, and cancer. We find that maintaining a volume at the recommended level throughout the follow-up is related to a greater risk reduction than engaging in sporadic high-volume activity in some periods mixed with inactivity. The trajectory analysis reveals that being physically active throughout middle adulthood is associated with a 10-28% lower incidence of major chronic diseases after age 60. This work demonstrates that maintaining physical activity over the long term provides added benefits beyond activity volume alone for chronic disease prevention.