The phylogenetic relationships of 14 species of Stilestrongylus were analyzed using the comparative morphology of 21 characters. We obtained 2 shortest trees of 50 steps, with a consistency index of 0.540 and 25 apomorphic character states. Ingroup monophyly was supported on these trees by 2 and 3 synapomorphies, respectively, and the ingroup was defined by the following characters: greater number of ventral ridges relative to the number of ridges dorsally, asymmetric bursa, and externodorsal rays differing in size. Sister-group relationships among Stilestrongylus and the other genera designated as outgroups are relatively consistent with those postulated by Durette-Desset's in a classification of the Nippostrongylinae. The ancestor of the species groups comprising Hassalstrongylus, Guerrerostrongylus, and Stilestrongylus originated and diversified in murids in the Neotropical region. Species of Hassalstrongylus occur in rodents between southeastern North America and the eastern part of South America (Brazil and Argentina), whereas species of Guerrerostrongylus and Stilestrongylus diversified exclusively in rodents in the Neotropics.