Healthcare workers have a high risk of burnout . This study aimed to investigate if the numbers of physical symptoms are associated with burnout among healthcare workers . We conducted a cross-sectional survey at a large university in Tokyo, Japan, in 2016 . Participants were 1080 : 525 faculties and 555 hospital workers . We investigated 16 physical symptoms perceived more than once per week and examined the association between the number of physical symptoms and Copenhagen Burnout Inventory (CBI); work-related (WBO), personal (PBO), and client-related (CBO) burnout . All CBI scores were higher among hospital workers than among faculties: WBO (43 vs. 29), PBO (50 vs. 33), CBO (33 vs. 29). Moreover, the higher the number of physical symptoms perceived, the higher the degree of burnout scores became (trend p-values <0.001), except for CBO among faculties . Job strain (all except for CBO among hospital workers) and work–family conflict were associated with an increased risk of burnout . Being married (WBO and CBO among faculties), having a child (except for PBO and CBO among faculties), and job support (faculty and hospital workers with WBO and faculties with PBO) were associated with a decreased risk of burnout . Multiple physical symptoms might be useful for identifying high risk individuals for burnout.