Islands often comprise unique faunal assemblages, particularly when they lie at the boundaries of different bioregions or host endemic taxa. Bats are among the few mammals that can regularly be found on islands around the world, yet knowledge on insular bat assemblages is often poor and anecdotal. Here we integrate different approaches to assess the bat assemblage composition on the Mediterranean island of Pantelleria. We found that at least six species occur on the island, including two typically African taxa, thus doubling the numbers known from previous studies. We provide insights into the distribution, biogeography and conservation planning of these bat species, highlighting the importance of studying and conserving insular bat populations.