The Himalayan region is a global biodiversity hotspot that faces severe pressures from a growing human population, rapid urbanization and climate change. While taxa like birds, plants and butterflies have been effectively surveyed along vast elevational gradients, there is immense paucity in such data for nocturnal and secretive animals like bats. We surveyed 22 sites in Uttarakhand in the western Himalayas spread across 600 m to 3,000 m a.s.l. A total of 35 species from 15 genera and five families were observed. We report the echolocation calls for 32 species, of which nine species have been recorded for the first time in the world. Additionally, we sequenced the cytochrome oxidase-I (COI) gene from 26 individuals belonging to 13 species. Eight species: Tadarida teniotis, Murina leucogaster, Murina aurata, Murina cyclotis, Myotis cf. frater, Myotis cf. annectans, Mirostrellus joffrei, Arielulus circumdatus and Eptesicus tatei are new records for the western Himalayan region. Myotis cf. frater is a new record for the entire Indian Subcontinent. In addition, we present an updated checklist of 49 species of bats for Uttarakhand state by reviewing previous literature. Our study will serve as a crucial baseline for future research by researchers and forest management authorities as the echolocation calls will be used to collect more occurrence data to elucidate the patterns of species richness across elevational gradients and to initiate the monitoring of bat populations.