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(1) Background: this study investigated the preventive measures implemented in the workplace and evaluated knowledge, attitudes and adherence behaviors regarding SARS-CoV-2 routes of transmission and preventive measures in a group of workers. (2) Methods: this cross-sectional study was conducted from May to July 2021 among 501 workers in the Campania region, in Southern Italy. (3) Results: 80.5% of respondents declared that their company had implemented the main COVID-19 preventive measures, and 54.7% of respondents knew SARS-CoV-2 routes of transmission along with the main preventive measures. Moreover, 34.2% were highly concerned about contracting SARS-CoV-2 in the workplace and transmitting it to family. Adherence to all preventive measures in the workplace involved 42.5% of respondents. The results of the multivariate logistic regression model revealed that significant determinants of adherence to all preventive measures in the workplace were being female, working not as a manager or office employee, cohabiting with someone that received a diagnosis of SARS-CoV-2 infection, knowing SARS-CoV-2 routes of transmission and the related main preventive measures, being highly concerned of contracting SARS-CoV-2 in the workplace and transmitting it to family and believing that COVID-19 vaccine offers high protection against disease. At the time of the survey, 47.5% of respondents had already received COVID-19 vaccine. Among unvaccinated respondents, 11.8% expressed unwillingness to get vaccinated for COVID-19. (4) Conclusions: These findings highlighted a good awareness about COVID-19 prevention and underlined a good propensity to get vaccinated among workers. Therefore, there is the need that preventive measures should be prioritized in the working context.