Bats have become an epidemiologically significant source of pathogenic microorganisms, such as leptospires, the causative agents of leptospirosis. However, little information exists about bats and their potential role as a reservoir of pathogenic Leptospira spp. in Colombia. The aim of this study was to evaluate the presence of pathogenic Leptospira spp. in the kidneys of bats from the Caribbean region of Colombia deposited in the collection of mammals of the Museo Javeriano de Historia Natural (MPUJ-MAMM).DNA was extracted from twenty-six kidney samples from a total of 13 species of bats captured in Colombia. First, 16S ribosomal RNA conventional PCR was performed to detect the presence of Leptospira spp. Then, in samples that tested positive, LipL32 PCR was performed to detect pathogenic Leptospira spp. by sequencing and phylogenetic analysis.The presence of Leptospira spp. was observed in 7/26 (26.9%) bats from the following 6 species: Carollia perspicillata, Glossophaga soricina, Dermanura phaeotis, Uroderma bilobatum, Desmodus rotundus, and Lophostoma silvicolum, and pathogenic Leptospira spp. were detected in 4/26 samples (15.4%).This study suggests that bats present in the Caribbean region of Colombia could be potential reservoirs of pathogenic Leptospira spp.