BACKGROUND: Total Worker Health METHODS: We conducted a comprehensive evaluation of a TWH leadership training program within a major Latin American agribusiness operating in Guatemala, Nicaragua, and Mexico. The evaluation followed best practices, encompassing behavior change and outcome assessment, including a post-training evaluation measuring training reactions, TWH leadership practices, training transfer intentions, modifiable health indicators, training transfer facilitators and barriers, and goal setting. A follow-up evaluation occurred three months post-training to assess the sustainability of TWH leadership practices, intentions, and health indicators. Linear mixed effects models were employed for analyzing training reactions. RESULTS: A total of 121 leaders (61% of all leaders) participated in TWH leadership training. The findings indicated consistently high training ratings across countries. Leaders reported positive leadership practices immediately post-training, and these improvements persisted three months later. Health indicators among leaders in different countries suggested a reduction in home and work-related stress and overall improvement in well-being. CONCLUSION: Positive changes in leadership practices and notable health improvements were observed. No significant differences were found among countries. This study contributes to understanding TWH implementation in Latin America, emphasizing the importance of tailoring programs to address specific organizational and cultural needs.