Between 2021 and 2024, Ecuador’s penitentiary system faced an unprecedented security crisis marked by deadly prison massacres linked to organized crime. Based on these unusual circumstances, the current research examined the working conditions and labor rights of prison security personnel and their biopsychosocial impacts. Framed within a comprehensive public security approach, following the March 29, 2023, reform to the Public and State Security Law, a mixed-methods strategy was used, being legal review, previous research, and digital surveys with officers from prisons of the Provinces of Cotopaxi, Guayas, and Azuay. Findings reveal that, despite a legal framework that guarantees labor rights, these are often not enforced. Staff members work under extreme violence, stress, and lack of proper equipment, training, and institutional support. This results in significant health effects such as chronic stress and anxiety. However, self-reporting on mental health issues was low, suggesting normalization or underestimation of psychological harm. The study underscores the urgent need for institutional reforms to enforce labor rights, enhance training, and provide psychosocial support, particularly during prison emergencies. Improving prison officers’ conditions is essential not only for their well-being but also for strengthening Ecuador’s broader public security system against organized crime threats.