The complete life cycle and developmental stages of the fluke, Metorchis ussuriensis sp. nov. (Trematoda, Platyhelminthes), are herein described. The results of the present experiments showed that, for flukes from the Primorsky Region in the Russian southern Far East, the first intermediate hosts are the snails Parafossarulus spiridonovi and Boreoelona ussuriensis, and the second intermediate hosts are freshwater fish, tadpoles, and snails. The definitive host in this experiment was Anas platyrhynchos dom. Morphometric parameters of M. ussuriensis sp. nov. demonstrate similarities with Metorchis taiwanensis, but the two species differ in the sizes of their bodies, sizes of suckers of adult worms, and sizes of cercariae, as well as respective positions of the finfold in cercariae. Phylogenetic reconstructions and genetic distances using the cox1 gene sequences support the conclusion that M. ussuriensis sp. nov. is well distinguished from all other species of the genus Metorchis, while sequences of internal transcribed spacers (ITS1 and ITS2) failed to separate M. ussuriensis sp. nov., Metorchis bilis, and Metorchis xanthosomus. In addition, we sequenced 1,402 bp of the 28SrRNA gene of M. ussuriensis sp. nov. being the first 28S sequences in the genus Metorchis. Comparison to other trematodes suggests that 28SrRNA could proof suitable for the differentiation of trematode species.
As a result of experimental studies conducted in the Russian southern Far East, adult worms from the genus Metagonimus were obtained. A comparative analysis of the morphometry of these worms with other Metagonimus representatives showed that they are most similar to M. katsuradai Izumi, 1935 and M. otsurui Shimazu & Urabe, 2002 found in Japan, due to the ratio of suckers and the positions of the testicle, uterus and vitellaria. However, Russian worms differ from species in Japan by other metric characters: they differ from M. otsurui by the maximum size of most organs and from M. katsuradai by body width, pharynx length, and maximum size of testes and ovary. At the same time, they are identical to a trematode from the Russian southern Far East, which was previously identified as M. katsuradai. The validity of this species was also confirmed by genetic data. According to the 28S gene and the internal transcribed spacer 2 (ITS2) region of rDNA, as well as the cytochrome c oxidase I (cox1) gene of mtDNA, the Metagonimus specimens found in Russia differ from published genetic data for other members of this genus. However, both morphological similarity and molecular data showed that M. pusillus sp. nov., M. katsuradai and M. otsurui are most likely cryptic species. Furthermore, additional data based on a mitochondrial marker were provided for M. suifunensis Shumenko, Tatonova & Besprozvannykh, 2017 from Russia.
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