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• Only 62.2% knew that hepatitis B and flu vaccines are recommended for them. • Only 13.2% knew all vaccines recommended for them. • High vaccination coverage are reported for hepatitis B, tetanus, and pertussis. • Only 9.2% received all recommended vaccines. Low rates of vaccine coverage have resulted in a resurgence of several vaccine-preventable diseases in many European countries. Routine vaccination of healthcare workers (HCWs) is important to reduce disease transmission, and to promote vaccine awareness and acceptance in the population. The objectives of this cross-sectional study were to investigate knowledge and beliefs about vaccines and to evaluate self-reported immunization coverage with vaccines recommended for HCWs. Additionally, the effects of several factors on these outcomes have been evaluated. A survey was conducted between September and November 2018 among a random sample of HCWs in cardiac, adult, and neonatal critical care units of 8 randomly selected hospitals across the Campania and Calabria Regions in Italy. Multivariate logistic and linear regression analysis has been performed. A total 531 HCWs returned the questionnaire for a response rate of 54.9%. Based on a vaccination knowledge score ranging from 0 to 9, more than half of the participants (55.4%) knew few of the vaccines recommended for HCWs (≤3 correct answers), 16.2% knew some vaccines (4–6 correct answers), and 28.4% knew most vaccines (≥7 correct answers), and only 13.2% knew all the vaccines recommended for HCWs. However, two-thirds (62.2%) knew that hepatitis B and influenza vaccines were recommended, and this knowledge was significantly higher among females (p < 0.001), among HCWs aged between 50 and 59 years (p = 0.01) compared with those aged < 30 years, and in those who search for information about recommended vaccines for HCWs (p = 0.012). The vaccine knowledge was significantly lower among nurses and nursing supporting staff compared with physicians (p = 0.032). Approximately two-thirds (62.7%) of HCWs considered themselves at risk of contracting vaccine-preventable infectious diseases during their professional practice. High rates of coverage were self-reported for hepatitis B (96.3%), tetanus and pertussis (93.7%), whereas they were lower for measles/mumps/rubella (80.5%), chickenpox (65.3%), and influenza (35.8%). Only 9.2% of HCWs reported prior receipt of all recommended vaccines. Male HCWs were less likely to report prior receipt of all recommended vaccines (p = 0.011). HCWs aged between 30 and 39 years compared with those aged < 30 years (p = 0.001) and those who knew some (p < 0.001) and most (p = 0.007) of all vaccines recommended for HCWs were more likely to self-report to be immunized. Additional training about the vaccinations is needed to improve HCWs knowledge and to address specific concerns which may lead to better uptake among this group. [ABSTRACT FROM AUTHOR]