The reef fish communities of the Guanahacabibes National Park have been studied for 20 years using various methodologies that have allowed us to understand aspects of their diversity and structure. However, due to gaps in information about the abundance and distribution of mesopredators (big fish and sharks), a new study was conducted in 2017 to determine their structure, explore the influence of different factors on their spatial variability, and evaluate their behavior. To achieve this, the Baited Remote Underwater Video Surveys (BRUVs) methodology was successfully applied, locating a single set of BRUVs at 90 sites distributed across 9 sectors of the park’s functional zoning. Variability in mesopredator metrics and their potential prey was assessed through a PERMANOVA analysis; a distance-based linear model (DISTLM) was used to explore the relationship between mesopredator abundance and biological, abiotic, and condition variables; and animal behavior was classified as incidental, cautious, or aggressive. A total of 64 fish species were identified, 7 of which were mesopredators, and 3 were sharks. An uneven distribution and abundance were observed among sectors, with the most abundant mesopredators being Carcharhinus perezi, Sphyraena barracuda, and Mycteroperca bonaci. Mesopredator abundance was more closely related to the condition of zone use and its regulations than to biological and abiotic variables. Sharks were more abundant in strictly protected areas, which coincided with relatively murky waters and stronger currents. More than 50% of the observed sharks displayed exploratory and aggressive behavior towards the bait basket. The analyzed metrics validate the effectiveness of the management of the protected area and suggest the presence of healthy and resilient mesopredator fish communities.