A new species of a predator aphelenchoidid, Seinura caverna n. sp., is described and illustrated. The new species was isolated from bat guano collected from a calcareous cave in Japan. The new species is characterised by its three-lined lateral field, secretory-excretory pore at the level of the posterior two-thirds of the metacorpus, a long post-uterine sac, hermaphrodite tail shape elongate conoid with a filiform tip, and an androdioecious reproductive mode. The new species is typologically and biologically close to S. steineri, but is distinguished by its longer post-uterine sac, slightly longer stylet and slightly larger median bulb. The comparisons with other morphologically similar species, i.e., S. chertkovi, S. tenuicaudata and S. steineri, are discussed. A molecular phylogenetic analysis based on two small and large subunit ribosomal RNA genes revealed that the new species is located at the basal position of clade 3 of the Aphelenchoididae, clearly separate from S. demani, its congener with unclear rectum and anus, which is located at the derived position, suggesting that further generic revision is necessary for the genus. Aphelenchoides lii n. comb. (=Seinura lii) is proposed.