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Introduction. The aim of the study was to evaluate the prevalence of cardiovascular risk factors (CVRF) and to analyse the influence of sex, work shift and job position on sleep quality and other CVRF. Methods. This was a multicenter prevalence study conducted in 2023 using a survey among nursing staff at three Spanish hospitals. Sleep quality was assessed using the Oviedo Questionnaire; nicotine addiction with the Fagerström Test; physical activity with the IPAQ questionnaire; adherence to the Mediterranean diet with the PREDIMED questionnaire; and perceived stress with the Cohen scale. Results. A total of 727 workers participated in the study. Rotating-shift workers (71.65%) exhibited poorer sleep quality (p<0.001), lower levels of physical activity (p<0.001), and higher perceived stress (p<0.001) compared to day-shift workers. Hypercholesterolemia (p<0.05) and hypertension (p<0.05) were more prevalent among day-shift workers. Women reported higher perceived stress (p<0.01) and poorer sleep quality (p<0.05) than men, whereas men had a higher prevalence of cardiovascular events (13%). Workers who had experienced a cardiovascular event or had higher perceived stress showed worse sleep quality (p<0.01). Conclusions. There is evidence that the negative health effects associated with shift work increase with age. Sleep quality, hypertension, and stress are particularly affected. Shift work and physical activity appear to negatively influence sleep quality, which is worse among rotating-shift workers.