Here I report a fossil microvertebrate fauna from a late Holocene cave deposit in northwestern Cuba. This study provides new chronological data for the understanding of the post-Pleistocene survival of some of Cuba's rarest extinct bats, and a post-Columbian record of Desmodus rotundus. Remains were excavated from a superficial, mound-like deposit with no dicernible stratigraphy. The bones of 13 bat species, one owl, one rail, a small colubrid snake, one frog, a nesophontid shrew, a rat, a mouse, and two capromyid rodents were extracted from the assemblage. The chiropterans were the most abundant and included the extinct taxa Desmodus rotundus and Artibeus anthonyi, the rare Lasiurus insularis,and the endangered Natalus primus. The mixed assemblage originated from accumulated owl pellets and carcasses of roosting bats. Three Artibeus jamaicensis scapulae from the same level and bone coloration as the Desmodus material yielded a modern radiocarbon content indicative of post-thermonuclear testing era (114.9 % ± 0.6 pMC), indicating an essentially modern age for the specimens. Material from the deepest level, however, is interpreted to be older due to factors such as a probable slow rate of accumulation, difference in bone coloration, and mineralization. These results support the survival of some extinct bat species well into the late Holocene of Cuba.