A new species of argasid tick, Ornithodoros (Alectorobius) rioplatensis Venzal, Estrada-Peña, & Mangold, is described from larval and adult specimens collected in Uruguay. Further specimens from Argentina, Paraguay, and Chile are ascribed to the new species. Key distinguishing characters of larvae of the species are the number of pairs of dorsal setae (19–20 but typically 20 in O. rioplatensis, compared with 16–17 but typically 17 in O. puertoricensis and O. talaje), and the length of the hypostome, which is clearly longer in O. rioplatensis than in O. talaje. Morphological details of the adults of these species are inadequate for taxonomic purposes. The genetic distance between O. puertoricensis and O. rioplatensis, based on 16S rDNA sequencing, is ≈12.7%. Variability in some morphological features, using multivariate morphometric discrimination to assess congruence between populations, was examined among larval specimens of O. talaje and O. puertoricensis collected from the United States, and Central and South America. Larval specimens of O. talaje from Guatemala (near the type locality of the species) were morphologically different from Mexican and southern U.S. populations originally described as O. talaje. In the absence of DNA data for these populations, and because of inadequate statistical discrimination among body characters, we prefer to refer to the Mexican and U.S. material as O. nr. talaje until further analysis of population variability is conducted using an adequate sample size.