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Most grouper species worldwide are threatened by overfishing. Effective marine protected areas (MPAs) are known to enable population recovery, and consideration of vulnerable species' reproductive behaviours is fundamental to monitoring and management plans. Many groupers produce sounds associated with reproductive behaviours. Recording these sounds helps to locate spawning sites and improve management efforts to ensure reproduction and viability. This study focuses on a poorly studied yet likely vulnerable grouper species, Mycteroperca rubra, providing novel insights into its reproductive biology by combining underwater visual census surveys, direct visual observations and passive acoustic monitoring within a Mediterranean MPA during two consecutive summers (2017 and 2018). Results indicate that M. rubra individuals were more abundant and larger at one of the protected study sites, where they also occasionally formed unusual aggregations (<30 individuals), likely for spawning. These aggregations and the observation of courtship behaviours suggest that M. rubra spawns in the surroundings of this study site. Moreover, grouper-like unknown sounds were recorded exclusively at this site, suggesting they are associated with M. rubra courtship behaviours. Therefore, this study provides a basis for validating M. rubra sound production and supports the monitoring of its spawning sites via passive acoustics to improve MPA conservation effectiveness. [ABSTRACT FROM AUTHOR]