In a scenario where escalating human activities lead to several environmental changes and, consequently, affect mammal abundance and distribution, β-diversity may increase due to differences among sites. Using the ecological uniqueness approach, we analyzed β-diversity patterns of ground-dwelling mammal communities recorded through comprehensive camera trap monitoring within eight tropical forests protected areas in Mesoamerica and South America under variable landscape contexts. We aimed to investigate whether the contribution of single sites (LCBD) and single species (SCBD) to overall β-diversity could be explained by community metrics and environmental variables, and by species metrics and biological traits, respectively. Total β-diversity was also partitioned into species replacement and richness difference. We related LCBD to species richness, total relative abundance, functional indices, and environmental variables (tree basal area, protected area size, NDVI, and precipitation seasonality), and SCBD to species naïve occupancy, relative abundance, and morphoecological traits via beta regression. Our findings showed that LCBD was primarily explained by variation in species richness, rather than relative abundance and functional metrics. Protected area size and tree basal area were also important in explaining variation in LCBD. SCBD was strongly related to naïve occupancy and relative abundance, but not to biological traits, such as body mass, trophic energy level, activity cycle, and taxonomic category. Local β-diversity was a result of species replacements and to a lesser extent differences in species richness. Our approach was useful in examining and comparing the ecological uniqueness among different sites, revealing the regional scale current status of mammal diversity. High LCBD values comprised sites embedded within smaller habitat extents, hosting lower tree basal areas, and harboring low species richness. SCBD showed that relatively ubiquitous species that occur at variable abundances across sites contributed most to β-diversity.