Molecular and morphological differentiation of two similar species of Accacoeliidae (Digenea): Accacladocoelium macrocotyle and A. nigroflavum from sunfish, Mola mola.

In a study of 106 sunfish, Mola mola (L.), from Mediterranean waters, 2,731 worms, belonging to 2 congeneric species of accacoeliids, Accacladocoelium macrocotyle (Diesing, 1858) Robinson, 1934 and Accacladocoelium nigroflavum (Rudolphi, 1819) Robinson, 1934 , were collected from the digestive system. It is often difficult to differentiate between these 2 species as they are sympatric and very similar; in fact, according to previous descriptions, the extent of the vitellarium is the only interspecific difference, described as extending posteriorly to the ovary in A. macrocotyle and as anterior to the anterior testis in A. nigroflavum. However, this diagnostic trait is not always valid; moreover, it is often indistinguishable because it is masked by the uterine eggs. Here, the morphology of new specimens of both species has been studied in detail and combined with molecular analysis. This study shows that the extension of the vitellarium is very similar in both species, but the degree of ramification differs. Furthermore, although the morphological differences are very slight, both species seemed genetically different: intraspecific differences ranged between 0.8 and 1.5% in internal transcribed spacer (ITS)-2 rRNA gene (ITS2) and between 0.5 and 1.6% in cytochrome c oxidase subunit 1 (COI) and interspecific differences ranged between 2 and 3.2% in ITS2 and between 9.6 and 10.6% in COI. In addition, we observed previously undescribed morphological differences, which help to differentiate these 2 species: the oral sucker is relatively smaller in A. nigroflavum than in A. macrocotyle and the ovary is usually relatively longer in A. nigroflavum than in A. macrocotyle.