Unexpected morphological and molecular diversity of trematode (Haploporidae: Forticulcitinae) parasites of mullets from the ocean Pacific coasts in Middle America.

Two new genera and four new species from subfamily Forticulcitinae are described from the intestines of white mullet (Mugil curema) and flathead grey mullet (Mugil cephalus) collected in 27 localities across a wide geographical range on Pacific Ocean slopes comprising three countries in Middle America: Mexico, Guatemala and Costa Rica. The new genera Ekuarhuni n. gen. and Overstreetoides n. gen. had to be erected to accommodate two new species, Ekuarhuni papillatum n. sp. and Overstreetoides pacificus n. sp., with unique morphological traits that differentiate them from the two genera described previously. In addition, two new species, Forticulcita minuta n. sp. and Forticulcita isabelae n. sp., were described, which were characterized as exhibiting a small body size (< 1100 μm long). These new species were classified as the diminutive morphotype, together with three other congeneric species of Forticulcita. Forticulcita minuta n. sp. is distinguished by being the smallest species within the genus (< 305 μm). Meanwhile, Forticulcita isabelae n. sp. is distinguished by its body size and testis length. In specimens of the four new species, sequencing was performed with two molecular markers, the large subunit (LSU) and the internal transcribed spacer 2 (ITS2) of nuclear rDNA, and the results were compared with other sequences available in GenBank. Phylogenetic analyses performed with the combined dataset of the two nuclear molecular markers (LSU + ITS2) placed all the analysed species within the clade of Forticulcitinae with strong bootstrap support (100%) and a high Bayesian posterior probability (1.0). The four new species showed differences in abundance in their definitive hosts and were widely distributed along the Pacific Ocean coasts of Mexico, Guatemala and Costa Rica, in Middle America.