This article presents attitudes and practices regarding COVID-19 vaccination in the South American population. The study collected data from a self-administered survey distributed through social media platforms between February and April 2022 (N = 6555). The survey included questions related to participants’ sociodemographic background, flu vaccination practices, sources of information about COVID-19, and opinions regarding pandemic management and vaccination against SARS-CoV-2. The respondents agreed with the statement that COVID-19 vaccines were necessary (86.4%), effective (79.8%), safe (79.1%), and should be mandatory (64%). Overall, 83.4% accepted vaccination and 12.3% refused it completely. Main rejection reasons were safety (65.8%) and efficacy (54.9%) issues, and rushed development and approvals (49.1%). Vaccine uptake was associated with being ≥60 years, being a healthcare worker, previous influenza vaccine uptake, adherence to preventive measures, the death of ≥1 close people from COVID-19, and being informed through mass media or health authorities’ channels. Vaccine uptake inversely correlated with male gender, low educational level, and use of closed social networks for COVID-19 information purposes. This study provides valuable insights into COVID-19 vaccination attitudes and practices in South America that may be used to promote vaccine uptake in the region. Higher COVID-19 vaccination acceptance among people with previously acquired prevention habits reinforces the importance of routine health promotion strategies.