The genus Nematostrigea comprises trematodes parasitising in fish-eating birds of Europe, Africa, and North America. Their life cycle is unknown. To provide the first description of metacercariae of Nematostrigea serpens serpens, a nominative subspecies of the type species of Nematostrigea, and to record metacercaria of Nematostrigea vietnamiensis, with molecular data. Encysted metacercariae of N. serpens serpens and N. vietnamiensis were collected from fish in Russia and Vietnam, respectively, and were processed, identified, and documented using standard morphological techniques. The 28S rRNA gene of metacercariae of both species and the cox1 gene of metacercariae an earlier studied adult of N. serpens serpens were sequenced and used for phylogenetic analysis. Metacercaria of N. serpens serpens have a spatulate body with a foliate forebody and a short hindbody, two long lateral rectilinear pseudosuckers, and the holdfast organ with bifurcated anterior and entire posterior external lobes. The analysis of the cox1and 28S rRNA gene sequences unequivocally showed the conspecificity of metacercariae and adult stage of N. serpens serpens. Based on the 28S rRNA gene sequence-based phylogenetic analysis, the genus Nematostrigea is a sister taxon to the group of strigeid genera Cotylurus + Ichthyocotylurus. Morphological and phylogenetic data demonstrated that N. vietnamiensis does not belong to the genus Nematostrigea and is possibly be affiliated with the crassiphialine trematodes. This is the first record of metacercariae of N. serpens serpens. N. vietnamiensis, renamed Prodiplostomulum vietnamiense comb. nov., must be moved to the crassiphialine ‘Prodiplostomulum’- type metacercariae group.