Venous thromboembolism (VTE) is the third most common type of cardiovascular disease. An association between high level of physical activity (PA) and the onset of VTE has been found in some, but not all previous studies. We aim to study the association between PA-level and VTE in a cohort of men with updated data on PA levels at four occasions. We used data from the Uppsala Longitudinal Study of Adult Men (ULSAM) study initiated in 1970, a study of men at age 50 years (n = 2,294 at baseline) examined on leisure time PA by questionnaire and traditional cardiovascular risk factors. Examinations were repeated at ages 60, 70, and 77, and follow-up was completed after a median time of 33 years. Cox regression analysis with hazard ratios (HRs) using updated covariates for PA and risk factors was performed on the association of PA levels with incident VTE, with adjustments for established cardiovascular risk factors (systolic blood pressure, LDL- and HDL-cholesterol, BMI, diabetes, and smoking). Totally 186 men experienced a VTE during follow-up of 68,263 person-years at risk. Individuals with the highest PA level had an increased relative risk of VTE, adjusted HR, 2.22 (95% CI 1.05-4.67), when compared to individuals with the lowest level of PA. In this cohort of men with a follow-up of 27 years, the risk of VTE was increased at the highest PA level. Findings indicate that there could be an increased VTE risk with higher PA level including strenuous activities.Graphical AbstractIn this cohort of men with a follow-up of 27 years, the risk of venous thromboembolism was increased at the highest level of physical activity