Forestry plantations represent about 4 % of the global land cover and demand for wood is steadily increasing worldwide. Impacts of forest plantations on biodiversity are controversial; forest plantations could positively influence biodiversity by producing a buffer zone between native forests and agriculture, while replacement of native forests with plantations could reduce biodiversity. Chile is one of the main producers of wood worldwide, and production is largely based on intensively managed monocultures of exotic tree species. Only a few studies have looked at the effects of forestry plantations on biodiversity in Chile, mainly focusing on pine plantations. The aim of this study was to characterize habitat use and richness of bats between native forests, eucalyptus plantations and grasslands in a biodiversity hotspot in southern Chile to determine how land use affects an important mammalian taxa. We found no difference in use or richness of bats in eucalyptus plantations versus native forests. Regional context within the larger Valdivian watershed (Andes, central valley, coastal range) had a stronger influence on bat activity and richness than land use type (native forest, plantation, grassland), with the Andean region being the most diverse and where most bat activity is concentrated. Our results suggest that the composition and structure of the surrounding landscape mosaic may be fundamental to determine the impacts of forestry and human land use on biodiversity.