Phyllodistomum spinopapillatum sp. nov. is described from the urinary bladder of the Oaxaca killifish, Profundulus balsanus Ahl (Profundulidae) in Rio Pueblo Viejo and Rio Santa Cruz, Oaxaca, southwestern Mexico. The new species is described based on evidence gathered from morphology, scanning electron microscopy (SEM), and partial sequences of the 28S rRNA gene. Diagnostic characters of the new species of Phyllodistomum Braun 1899 are the presence of spines on the entire body surface and having a ventral sucker almost half the size of oral sucker. The new species possess a large number of dome-like papillae on the body surface with scattered distribution along the hindbody, and these papillae are characteristically spinulated. Phyllodistomum spinopapillatum sp. nov. most closely resembles P. inecoli Razo-Mendivil, Perez-Ponce de Leon and Rubio- Godoy, 2013, a parasite of the twospot livebearer Pseudoxiphophorus bimaculatus (Heckel) from Veracruz, in the Atlantic slope of Mexico. In addition to the new species, specimens of P. inecoli were also found parasitising the urinary bladder of cyprinodontiforms such as the Mexican molly Poecilia sphenops Valencienes in a pond at Santa Maria Coyotepec, and in Profundulus sp. in Rio Templo, both in Oaxaca, and in the Porthole livebearer Poeciliopsis gracilis (Heckel) in Rio San Juan, as well as in Profundulus punctatus (Gunter) from Rio Nueva Francia, both in Chiapas. The distribution and host range of P. inecoli is extended to freshwaters of the Pacific slope of Mexico, and to other cyprynodontiforms.