BACKGROUND: Clinical guidelines recommend physical activity (PA) to prevent diabetic peripheral neuropathy (DPN). However, these recommendations lack specificity in terms of optimal PA intensity and duration. We aimed to determine associations between PA, DPN and painful DPN (pDPN). METHODS: In a cross-sectional study of persons with diabetes, PA was assessed with a wrist-worn accelerometer, recording time spent at different milli-gravity (mG) intensity levels with categories set as light (45-100 mG), moderate (100-400 mG), or vigorous (>400 mG). DPN was defined by Michigan Neuropathy Screening Instrument questionnaire and pDPN by presence of bilateral foot pain. RESULTS: In 2878 participants (mean (SD) age: 58.6 (7.1) years, 39.1% female), time spent above 75 mG decreased DPN odds (OR, 95% CI) (0.97, 0.95 to 0.997). Further, more time spent performing moderate (0.95, 0.91 to 0.99) or vigorous (0.40, 0.22 to 0.70) PA decreased DPN odds. Overall, PA did not affect pDPN; however, among females, more time spent above 50 mG (0.92, 0.87 to 0.98) decreased pDPN odds. CONCLUSIONS: Increased PA decreased DPN likelihood, and in females, pDPN likelihood. Whether lack of PA occurs secondary to the presence of DPN or contributes to DPN remains to be determined.