Pseudonannolene spelaea n. sp. is the first strictly cave-dwelling species described for the family Pseudonannolenidae. It is found in iron ore caves in the Brazilian Amazon. The family Pseudonannolenidae is exclusively Neotropical and frequently found in caves of Brazil, from which 20 species are known. The new species is compared with its congeners and with related cave-dwelling species. The family Pseudonannolenidae is discussed, and comments are presented on the conservation status of the caves where the species is found, which potentially may be the target of anthropogenic impacts resulting from iron ore extraction.