This study aimed to assess health literacy (HL), preventive behaviors (PBs), and the factors associated with PBs toward COVID-19 among village health volunteers (VHVs) and residents in urban and rural areas in the upper south of Thailand. A cross-sectional study was conducted using valid questionnaires administered using an online platform. Data were analyzed using descriptive statistics, t-tests, and binary logistic regression analysis. A total of 846 people participated in this study, and 62.8% of participants had an adequate level of HL. Furthermore, 55.0% of participants had an adequate level of PBs. Among VHVs, PB levels were adequate, whereas among residents, they were inadequate. Moreover, among VHVs, underlying diseases (adjusted odds ratio (OR): 1.77, 95% CI: 1.13–2.76), access to HL (adjusted OR: 4.28, 95% CI: 1.42–12.88), appraisal of HL (adjusted OR: 3.09, 95% CI: 1.90–5.02), and application of HL (adjusted OR: 2.51, 95% CI: 1.52–4.12) were associated with COVID-19 PB. Additionally, among residents, only marital status (adjusted OR: 1.59, 95% CI: 1.04–2.44), appraisal of HL (adjusted OR: 1.78, 95% CI: 1.18–2.84), and application of HL (adjusted OR: 3.85, 95% CI: 2.39–6.20) were associated with PBs towards COVID-19. Furthermore, individuals with inadequate HL demonstrated noteworthy significance in all dimensions of PBs. To prioritize and optimize preventive programs aimed at enhancing PBs, the government must take proactive initiatives in implementing robust health communication strategies across diverse media platforms, both in the present and future.