The Paraguaná moustached bat, Pteronotus paraguanensis (Mormoopidae), is one of the four species of bats endemic to Venezuela. Besides having a geographic distribution restricted to the Paraguaná Peninsula, it is currently classified as Endangered in the IUCN Red List of Threatened Species. Changes in land use, frequent human disturbance at diurnal roosts and poisoning with agro-pesticides from farming operations are some of the main threats that affect this species. Despite its conservation status and the pressing need to protect its feeding areas, habitat use of this species has not been well documented. The aim of this study was to record this bat's activity using acoustic monitoring to identify the vegetation types most frequently used throughout the year. We recorded echolocation calls over the course of eight months between 2015 and 2016, using walking transects distributed along the vegetation types most representative of the peninsula. Activity of P. paraguanensis differed significantly among vegetation types, reaching highest relative activity in thorn woodlands (45%), followed by premontane woodlands (20%), disturbed habitats (19%), thorn scrubs (11%), and columnar cactus forests (4%). Activity (mean number of bat passes/h) was comparatively higher in premontane woodlands (78.17 ± 49.82 SD) than in the other vegetation units (range: 4.39 ± 5.71 SD – 15.51 ± 16.11 SD) where this species was detected. This result is indicative that this species is mainly associated with forest habitats; however, it can also be present at lower frequency in disturbed lands. Our bat call recordings also help confirm that the Paraguaná Peninsula's isthmus represents an ecological barrier that precludes dispersion of P. paraguanensis to the mainland. Based on our findings, we highlight the need to focus conservation actions for this species on protection of the remnant patches of forest vegetation still present in the peninsula.