Background: The deterioration of Venezuela's health system in recent years undoubtedly contributes to an increased impact of the COVID-19 pandemic. Understanding healthcare workers' (HCWs) knowledge, attitudes, and practices (KAPs) toward COVID-19 in the early stages of the pandemic could inform their medical training and improve their preparedness.Methods: A online national cross-sectional survey was conducted between May 26th and May 30th, 2020, to assess KAPs among HCWs in Venezuela.Results: A total of 1,441 HCWs from all 24 regions of the country responded to the survey. The mean age of the HCWs was 44 (SD [standard deviation] 14) years; most were women (66.4%). Most HCWs were specialized doctors (48%), followed by nurses (13%) and resident doctors (12.3%). The majority of HCWs had good knowledge (76.3%), obtained information mainly from scientific literature (85.4%); had negative attitudes (53.6%), felt uncomfortable with their work during the current pandemic (59.8%); and reported appropriate practices (76.9%). However, participation in COVID-19 related training was absent in more than half of the HCWs. Positive attitudes were significantly more frequent in frontline workers than in non-frontline workers (p = 0.001). Bioanalysts, students, and doctors were more likely to have good knowledge; participating in training was a predictor for positive attitudes and older age was an appropriate practice predictor.Conclusions: HCWs, knowledge in Venezuela could be improved by strengthening education and training programs. Strategies should focus on reducing fear and improving attitudes toward the care of COVID-19 patients, as well as the promotion of preventive practices.