The application of genetic techniques has allowed to identify a number of cryptic species of bats and to better define their distribution ranges and conservation status. Long-eared bats Plecotus spp. occur throughout Eurasia and Africa, with a high number of morphologically similar species, often occurring in sympatry. Therefore, disentangling the distribution of one species from another may be challenging. Italy represents a diversity hotspot for long-eared bats in Europe, hosting six species belonging to the genus Plecotus. In this study we report on both unpublished ad-hoc investigations on the Alpine long-eared bat Plecotus macrobullaris by integrating field and molecular methods, as well as opportunistic verified records from citizen scientists, to redefine the distribution of this species in Italy. We retrieved a total of 27 unpublished confirmed records, namely 11 from iNaturalist, 4 from Facebook, and 13 from personal communications from bat workers. In Italy, the species was known to occur throughout the Alpine and Pre-Alpine chain, in some plain sites in Northeastern regions, and near the sea in Liguria. Based on new records, we proved that it occurs also in the other areas, including the Northern Apennine ridge. Thus, from the legal point of view, our findings also included a novel administrative region (Tuscany) within the area of presence of P. macrobullarisi.e., expand the responsibility of the species monitoring and implementation of conservation measures to additional authorities, as expected by the Habitats Directive.