Conversion of natural land covers to agriculture is a major cause of the global biodiversity decline. Bats are an important component of biodiversity in agricultural landscapes because they provide pest control services. Although management recommendations towards the enhancement of insectivorous bat populations in agro-ecosystems have previously been highlighted, little information is available for promoting bat conservation within viticultural landscapes. In the present study, we examined the role of the adjacent habitat on bat activity in vineyards of central Chile. We also evaluated differences in bat activity between the edges and the interiors of the vineyards in relation to the type of adjacent habitat. To accomplish this, we conducted acoustic surveys along edge and the interior of 16 vineyards bordering different adjacent habitats. Overall bat activity in vineyards was not influenced by the adjacent habitat type, but it was by the location within the vineyard; edges showed significant higher activity than the interior of the vineyards. Vineyards adjacent to native vegetation showed the highest levels of activity for Lasiurus varius, Lasiurus villosissimus and Myotis chiloensis compared to those adjacent to monoculture or urban areas. All bat species were most active at the edges of the vineyards as compared to the interior, which increase the probability of this group providing ecosystem services in vineyards. Therefore, vineyard edges, in particular those adjacent to native vegetation, should be considered as part of agricultural management in order to promote bat diversity and abundance in this crop.