Background: Like other health professionals, community pharmacists are exposed to stress factors (being efficient, avoiding mistakes and bearing emotional load), but they are also under the pressure of entrepreneurial responsibilities. The main objective was to assess the level of work-related stress in French community pharmacies. The other objectives of the study were to assess the associated comorbidities and causes of work-related stress. Methods: This observational cross-sectional study was sent to all French community pharmacies by email. The survey was anonymous and designed to collect the following items: socio-demographic factors, professional status, characteristics of community pharmacy, work-related stress (visual analogic scale-VAS), fatigue (VAS), sleep disturbances (questions), anxiety and depression symptoms (hospital anxiety and depression scale), medical consultation for work-related stress, medication use for work related stress, psychoactive drug-use and causes of work-related stress. Participants were included in the survey if they were pharmacists (owner or assistant) or pharmacy technicians working in a community pharmacy at the time of the survey. Exclusion criteria were defined as follows: pharmacy students or other professionals involved in a community pharmacy (e.g. dietician, beautician) and lack of professional status information. There was no age limitation. Results: After three months of data collection, 1,339 participants answered the survey and 1,272 participants were included in conformity with the inclusion and exclusion criteria, and to avoid missing data on the primary endpoint. Work-related stress was detected in 32.8% (417/1,272) of individuals (scores ≥70/100). Men were significantly more affected than women and there was no difference between professional statuses and no relation with the age of the participants. Work-related stress was significantly associated with anxiety, depression, fatigue, sleep disturbances, medical consultations, medication use, alcohol consumption above the WHO recommendations for men and psychoactive drug use. Three causes of stress were clearly identified and related to stress levels, workload, working atmosphere and deterioration of work quality. However, causes of work-related stress were significantly different among professionals, for example: entrepreneurial burden for pharmacists-in-charge and workload for employees (assistant pharmacists and pharmacy technicians). Discussion: Work-related stress has a very strong impact in French community pharmacies. This stress was associated with several comorbidities and induces health resource consumption. Several causes of work-related stress have been identified such as workload, working atmosphere and deterioration of work quality; however, these causes could be detected and managed to improve stress levels. We recommend developing individual and organizational stress management in French community pharmacies.