Human activities can substantially alter forest structure, consequently affecting forest animal communities. An important effect on bat behavior is given by canopy discontinuities like clearings or trails: while the former act as foraging sites, the latter are known to be used by bats to move inside the forest. For this reason, we can expect trails to have a strong influence on bat activity, with differences determined by the species' flying abilities. Here we assess the impact of trails on a local bat community by quantifying the effect of trails’ characteristics on the activity of bat species in relation to their wing morphology. We collected paired acoustic data and environmental variables along trails and in internal areas of a forest in central Italy. We modelled the activity levels of three bat guilds differing in wing morphology and identified the species mostly responsible for the observed compositional dissimilarity in between trails and internal zones by adopting a multivariate approach. Trail width and tree’s diameter were the main drivers of the observed differences between bat activity along trails and internal areas, but their effect differed among bat guilds. Edge and open-space foragers increased their activity along wider trails, while closed-space foragers showed an opposite trend; the latter also avoid trails in favor of internal areas especially when trees are larger. Four species yielded a significant contribution to the dissimilarity in activity levels between trails and internal areas, and namely Pipistrellus pipistrellus, Nyctalus noctula, Miniopterus schreibersii and Barbastella barbastellus. Our results show a clear effect of forest trails on bat activity, highlighting differences across functional groups in relation to trail characteristics. Furthermore, within guilds, not all species respond with the same intensity, suggesting differences in how different species are attracted to linear features and forest gaps. Overall, our results depict a complex interaction between forest trails and bat activity, suggesting that structural changes in forests can trigger diverse responses in bats. Future research on the topic may focus on assessing how such effects can affect bat communities at the landscape scale and longer time-scales.