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Background  Leeches (Hirudinida) play a significant role as intermediate hosts in the circulation of trematodes in the aquatic environment. However, species richness and the molecular diversity and phylogeny of larval stages of strigeid trematodes (tetracotyle) occurring in this group of aquatic invertebrates remain poorly understood. Here, we report our use of recently obtained sequences of several molecular markers to analyse some aspects of the ecology, taxonomy and phylogeny of the genera  Australapatemon  and  Cotylurus , which utilise leeches as intermediate hosts.    Methods  From April 2017 to September 2018, 153 leeches were collected from several sampling stations in small rivers with slow-flowing waters and related drainage canals located in three regions of Poland. The distinctive forms of tetracotyle metacercariae collected from leeches supplemented with adult Strigeidae specimens sampled from a wide range of water birds were analysed using the 28S rDNA partial gene, the second internal transcribed spacer region (ITS2) region and the cytochrome  c  oxidase (COI) fragment.    Results  Among investigated leeches, metacercariae of the tetracotyle type were detected in the parenchyma and musculature of 62 specimens (prevalence 40.5%) with a mean intensity reaching 19.9 individuals. The taxonomic generic affiliation of metacercariae derived from the leeches revealed the occurrence of two strigeid genera:  Australapatemon  Sudarikov, 1959 and  Cotylurus  Szidat, 1928. Phylogenetic reconstructions based on the partial 28S rRNA gene, ITS2 region and partial COI gene confirmed the separation of the  Australapatemon  and  Cotylurus  clades. Taking currently available molecular data and our results into consideration, recently sequenced tetracotyle of  Australapatemon  represents most probably  Au. minor ; however, unclear phylogenetic relationships between  Au. burti  and  Au. minor  reduce the reliability of this conclusion. On the other hand, on the basis of the obtained sequences, supplemented with previously published data, the metacercariae of  Cotylurus  detected in leeches were identified as two species:  C. strigeoides  Dubois, 1958 and  C. syrius  Dubois, 1934. This is the first record of  C. syrius  from the intermediate host.    Conclusions  The results of this study suggest the separation of ecological niches and life cycles between  C. cornutus  (Rudolphi, 1808) and  C. strigeoides/C. syrius , with potential serious evolutionary consequences for a wide range of host–parasite relationships. Moreover, phylogenetic analyses corroborated the polyphyletic character of  C. syrius , the unclear status of  C. cornutus  and the separate position of  Cotylurus raabei  Bezubik, 1958 within  Cotylurus . The data demonstrate the inconsistent taxonomic status of the sequenced tetracotyle of  Australapatemon , resulting, in our opinion, from the limited availability of fully reliable, comparative sequences of related taxa in GenBank.

parasites and vectors

Leeches as the intermediate host for strigeid trematodes: genetic diversity and taxonomy of the genera Australapatemon Sudarikov, 1959 and Cotylurus Szidat, 1928