The cross-sectional study assessed the knowledge, attitudes, and practices regarding the recommended vaccinations and factors affecting such outcomes among a sample of healthcare workers (HCWs) in public hospitals in Italy. Only 14.1% knew all the recommended vaccinations for HCWs. Physicians and those who had received information about vaccinations from scientific journals, educational activities, or professional associations were more likely to have this knowledge, while those aged 36–45 were more likely to have less knowledge than those in the age group below 36 years. Only 57.3% agreed that the information received about vaccinations was reliable. Respondents who had children, who worked in pediatric/neonatal wards, who were more knowledgeable, or who did not need further information about vaccinations considered the available information to be reliable. Only 17.7% of respondents always recommended vaccinations to their patients. This behavior was more likely to occur in physicians, in HCWs, in pediatric/neonatal wards, in those who considered the information received about vaccinations reliable, and in those who considered themselves to be at high risk of transmitting an infectious disease to their patients. Health promotion programs and efforts are needed to improve the level of knowledge about vaccinations and immunization coverage among HCWs.