Adults of the genus Posthodiplostomum, Dubois, 1936 are parasites of fish-eating birds, mainly of the family Ardeidae, and are globally distributed. The genus currently comprises 35 species, although recent molecular evidence has shown that the diversity of the genus is underestimated since several candidate species have been recognized. In the Neotropical region of Mexico, at least 6 Posthodiplostomum lineages have been detected with metacercaria stages recovered from unrelated fish hosts. Here, we obtained adult specimens of Posthodiplostomum from 6 fish-eating birds representing 2 families (Butorides virescens, Ardea herodias, Nycticorax nycticorax, Tigrisoma mexicanum – Ardeidae, and Rynchops niger and Leucophaeus atricilla – Lariidae) from 4 localities in southern Mexico. Specimens were sequenced for 2 nuclear (28S and ITS1–5.8S–ITS2) and 1 mitochondrial (cox1) molecular marker. Phylogenetic analyses allowed us to link metacercariae and adult specimens and recognized a lineage, which was described morphologically. The new species can be distinguished from its congeners by its prosoma morphology and body size; this is the first described species in the Neotropical region of Mexico. Additionally, new host and locality records for P. macrocotyle and P. pricei are presented, expanding their geographical distribution range in the Americas.