A new heterophyid species, Ascocotyle (Ascocotyle) nunezae n. sp., is described from adults found in the intestine of naturally infected heron, Casmerodius albus (type host), from the coastal lagoon of Celestún. Yucatan, Mexico, and a domestic chick (Gallus gallus), experimentally infected with metacercariae from Cichlasoma octofasciatum. The new species is characterized mainly by the number (32-37) and arrangement of circumoral spines, which form I complete row of 25-27 circumoral spines and 6-10 accessory spines on the dorsal side, and by the morphology of the ventrogenital sac with a large gonotyl, consisting of 2 indistinctly separated lobes of vesicular tissue. Ascocotyle (A.) nunezae is placed into the nominotypical subgenus Ascocotyle because of the presence of uterine loops at the pharyngeal region and position of vitelline follicles. However, it differs distinctly from other members of this subgenus by the presence of long intestinal ceca reaching posterior to the ventral sucker. Cichlids of the genus Cichlasoma from cenotes, lakes, and the river Río Hondo in the Yucatan Peninsula were natural second intermediate hosts of A. (A.) nunezae, with metacercariae encysted on their gills. Cichlasoma meeki (Brind) was the most heavily infected fish host (total prevalence 75%; mean intensity 11 +/- 9).