A new species of Alloglossidium is described from the intestines of 2 madtom species (Noturus leptacanthus and Noturus gyrinus) that were collected from the run of a small, unnamed spring system that drains into the Santa Fe River, Florida. Alloglossidium floridense n. sp. is morphologically very similar to other nonprecocious Alloglossidium spp. that use ictalurids as definitive hosts, but can be distinguished by a combination of its smaller overall size (length and width), large eggs in relation to its small body size, position of the vitellaria, ovary shape, and position of the ovary in relation to the cirrus sac. A comparison of nuclear rDNA sequences (spanning partial 18s, complete ITS1, 5.8s, ITS2, and partial 28s regions) showed that A. floridense n. sp. diverged by 0.70–3.17% from the other 4, nonprecocious species that infect ictalurids (Alloglossidium corti, Alloglossidium fonti, Alloglossidium geminum, and Alloglossidium kenti). The new species of Alloglossidium, described herein, is the first of the genus to be reported from Florida and the first to be recorded from N. leptacanthus. In light of the subtle morphological differences among the nonprecocious species that infect ictalurids, we discuss how previous descriptions of species traits that are not supported with genetic data are difficult to interpret because of the possible past nonrecognition of distinct species.