Three new species of Creptotrema (Trematoda, Allocreadiidae) with an amended diagnosis of the genus and reassignment of Auriculostoma (Allocreadiidae), based on morphological and molecular evidence.

Diversity of Creptotrema was investigated using morphological and molecular tools, including data for Creptotrema creptotrema (type-species). Three new species, parasites of Brazilian fishes, are described: Creptotrema conconae n. sp. (type-host, Imparfinis mirini Haseman), Creptotrema schubarti n. sp. (type-host, Characidium schubarti Travassos) and Creptotrema megacetabularis n. sp. (type-host, Auchenipterus osteomystax (Miranda Ribeiro)). The diagnosis of the genus was amended to include new features. The new species differ from each other mainly in terms of body shape, relative sucker size, and testes position. DNA sequences were obtained from Creptotrema spp. from Brazil, including 28S, ITS and COI. Genetic divergences among the new species and C. creptotrema varied from 2.1 to 5.2% (21-49 bp) for 28S, and 6.6 to 16.4% (21-45 bp) for COI. Phylogenetic analysis (28S) placed the newly generated DNA sequences of Creptotrema in a clade (Creptotrema clade sensu stricto) composed of C. creptotrema, the new species described herein, and all species previously described as Auriculostoma, revealing that Auriculostoma is best interpreted as a synonym of Creptotrema based on the principle of priority of zoological nomenclature. Creptotrema funduli, the single sequence of the genus previously available, was not grouped within the Creptotrema clade sensu stricto, suggesting the need for reevaluation of the taxonomic status of this species. Our results showed that Creptotrema represents a monophyletic genus of trematodes widely distributed across the Americas, which currently comprises 19 valid species of parasites of teleosts and anurans.