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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.

The systematic position and validity of species within genus Syngamus have always been controversial. In this present work, we evaluated the taxonomic status and phylogenetic relationships between three species of Syngamus nematodes ( S yngamus trachea , Syngamus taiga and Syngamus merulae ) and one taxa, determined only to the generic level, collected from respiratory tracts of passeriform hosts from Central Europe using newly obtained sequences of 2 nuclear markers (internal transcribed spacers 1 and 2 [ITS1, ITS2]) and a fragment of mitochondrial cytochrome oxidase I [ COI ] gene. Our results clearly showed that S. trachea , S. taiga and S. merulae are valid, molecularly and morphologically distinct species. Moreover, molecular analysis of adult female of Syngamus sp. collected from tracheae of the European robin Erithacus rubecula clearly indicate that these comprise separate species. In the derived phylogeny, the Syngamus clade is divided into two sub-clades: one comprised Syngamus species with a characteristic, well-developed cuticular collar around the oral opening ( S. trachea and S. taiga ) and a second that groups taxa without or with rudimentary collar ( S. merulae and currently sequenced Syngamus sp. from Erithacus rubecula ). These results clearly suggest that the degree of collar development (well-developed vs. rudimentary/absent) may be an important phylogenetic feature for determining the structure of the genus Syngamu s on subgeneric level. Additionally, our results support historical division of the genus Syngamus into two subgenera S. ( Syngamus ) and S. ( Ornithogamus ). [ABSTRACT FROM AUTHOR]

The genus Brachylecithum was for the first time subject to molecular taxonomic phylogenetic analysis in order to ascertain relationships among its component taxa. We used two markers-the nuclear ribosomal 28S ribosomal DNA (rDNA) gene and the mitochondrial cox1 gene, for six species of the genus; 11 sequences of partial 28S rDNA and partial cox1 were obtained from adult B. capilliformis, B. glareoli, B. kakea, B. laniicola, B. lobatum, and B. strigis, and from larval stages obtained from snails of the genus Cepaea. We propose to synonymize B. strigis with B. lobatum, while the genetic differences in the 28S rDNA gene and mitochondrial cox1 gene confirm the species status of B. capilliformis and indicate a distinct group within Brachylecithum, including B. kakea and B. laniicola. Cercarial and metacercarial isolates from the snails showed 100 % similarity to B. lobatum; thus, it is the first record of Cepaea snails as intermediate hosts of this species and the first report on life cycle abbreviation within the Dicrocoeliidae. [ABSTRACT FROM AUTHOR]

The systematic position of the Collyricloides massanae, a rare cyst-dwelling parasite, located on intestinal wall of European birds and rodents, have always been controversial. Based on newly obtained sequences of the 28 sDNA of C. massanae from avian and rodent host from Central Europe, and on the previously published sequences of several genera and families among Microphalloidea, we evaluate its taxonomic position and the phylogenetic relationships within the genera Collyriclum Kossack, 1911 and Collyricloides Vaucher, which form the family Collyriclidae Ward, . In the cladogram, C. massanae appears among the Pleurogenidae, forming a clade with Gyrabascus amphoraeformis (Modlinger, 1930) and Cortrema magnicaudata (Bykhovskaya-Pavlovskaya, 1950). We reject the commonly accepted placement of Collyricloides as the sister genus to Collyriclum within the Collyriclidae. Besides, we present and discuss the unusual records of C. massanae in the bank vole Myodes glareolus from northeastern Poland. [ABSTRACT FROM AUTHOR]

Genus Mesocestoides is a representative of the small cyclophyllidean family Mesocestoididae that is found parasitizing the small intestine of carnivores. The life cycle of cestodes from this genus is complex and requires two intermediate hosts. Cysticercoids are produced in the first intermediate host (oribatid mites), which when eaten by the second intermediate host (mainly rodents, but also other mammalian species, birds, reptiles, or amphibians) form tetrathyridia in the body cavity. Because of the rich history of nomenclatural evaluation of Mesocestoididae, the taxonomic status within the genus Mesocestoides is still unclear. Additional problem constitute the difficulty or even the impossibility in the determination of tetrathyridia based on morphological features. Thus, the aim of our study was to identify a molecular characteristic of the isolates of Mesocestoides from the second intermediate hosts (rodents) based on nuclear and mitochondrial ribosomal DNA data. We choose to analyze metacestodes isolated from two species of rodents ( Apodemus agrarius and Myodes glarolus) from different sites. As a result of amplification of 18S rDNA, we obtained partial sequences from four isolates ranging from 1,116 to 1,162 bp. In relation to mitochondrial sequence, 354 bp product of 12S rDNA was obtained from one isolate. The neighbor joining and maximum parsimony trees were constructed in order to examine the phylogenetic relationship within Mesocestoides spp. occurring in rodents from central Europe. The results of our research on the larval stages from rodents, living in a periphery of urban agglomeration as well as in an area of reserve protection, confirm the data of more frequently occurring Mesocestoides litteratus. [ABSTRACT FROM AUTHOR]