OBJECTIVE To investigate relationships between epicardial fat volume and atherosclerosis in multiple major vessel beds. RESULTS From the population-based Rotterdam Study, 2298 participants underwent computed tomography examinations to quantify epicardial fat volume and atherosclerotic calcification volume in the coronary arteries, aortic arch, and extracranial and intracranial internal carotid arteries. Using linear regression modelling, we investigated relationships of epicardial fat volume with atherosclerotic calcification volume in each vessel bed, adjusting for conventional cardiovascular risk factors (waist circumference, systolic and diastolic blood pressure, total and high-density lipoprotein cholesterol, smoking, diabetes, and usage of blood pressure-lowering and lipid-lowering medication). To test whether associations of epicardial fat with calcification per vessel bed were independent of calcification elsewhere, we created a model in which all vessel beds were entered together. We found that a larger epicardial fat volume was associated with larger calcification volumes in the coronary arteries, aortic arch, and extracranial carotid arteries in both sexes. After adjustment for cardiovascular risk factors, larger epicardial fat volume was related to coronary and extracranial carotid artery calcification volume in males only [difference in calcification volume per SD increase in epicardial fat volume: 0.12 (95% confidence interval, CI: 0.04; 0.19) and 0.14 (95% CI: 0.06; 0.22)]. These associations remained unchanged after entering all vessel beds into one model. CONCLUSIONS Larger volumes of epicardial fat are associated with larger amounts of coronary and extracranial carotid artery atherosclerosis in males, independent of cardiovascular risk factors. This could imply that epicardial fat also exerts a systemic effect on atherosclerosis development. Future longitudinal research is warranted to further disentangle these relationships with a specific focus on sex differences.