Partial 18S rRNA gene sequences of the three trichodinids, namely Trichodina modesta Lom, 1970, Trichodina paraheterodentata Tang and Zhao 2012. and Trichodinella epizootica (Raabe 1950) Šrámek-Hušek, 1953, were acquired and used to construct phylogenetic trees. The results revealed that Trichodinella epizootica clustered with Trichodinella sp.; Trichodina paraheterodentata Tang and Zhao 2012 was sister to the clade composed of Trichodina heterodentata Duncan, 1977 and Trichodinanobilis Chen, 1963; Trichodina modesta Lom, 1970 clustered with Trichodina reticulata Hirschman and Partsch, 1955. The branching order of species within the Mobilia clade was closely correlated with GC content. Furthermore, blade morphology was also found to be the primary morphological character in determining the phylogenetic relationships among members of the genus Trichodina. The present findings suggest that the genus Trichodina is paraphyletic when species of Trichodinella are included in the analyses.
In the present study, we isolated three populations of Myxobolus ampullicapsulatus from the gills of crucian carp, Carassius auratus auratus, two from Yongchuan, Chongqing area and one from Poyang Lake, Jiangxi area, China, sequenced their complete small subunit ribosome RNA gene, analyzed their genetic distance and gene similarity, and explored their relationship based on Bayesian inference and maximum likelihood analyses of their small subunit ribosomal DNA. The results combined with their morphological characteristics suggest that M. ampullicapsulatus infecting the gills and pharynx of allogynogenetic gibel carp, Carassius auratus gibelio, should be Myxobolus honghuensis. This study highlights the importance of DNA sequence comparisons for distinguishing Myxobolus species and indicates that the intra-species identification for the two Myxobolus species mentioned in the present research should be less than ten variation sites. In morphology, M. honghuensis Liu et al. (2012) parasitic on the gills of C. auratus auratus (goldfish) was collected from Chongqing area, and its mature spore was 16.5–19.5 × 8.5–10.0 μm in size, polar capsule was 7.0–10.0 × 2.5–4.0 μm in size, and polar filament had 9–10 coils. M. honghuensis Liu et al. (2012) isolated from the pharynx of C. auratus gibelio was sampled in Hubei area, and its mature spore was 15.1–19.5 × 9.0–11.3 μm in size, polar capsule was 7.9–8.1 × 3.0–4.5 μm in size, and polar filament had 7–8 coils.
It is difficult to differentiate similar trichodinids solely based on morphological examination, thus other identification methods, such as molecular identification, are necessary for identification. One mobilid ciliate named Trichodina pseudoheterodentata sp. n. was isolated from the gills of channel catfish, Ictalurus punctatus, in Chongqing, China. In the present study, its SSU rDNA was sequenced for the first time. Based on the results from both morphological identification and SSU rDNA sequencing, the new species was identified and compared with similar species. The morphological analysis revealed that T. pseudoheterodentata is a large Trichodina species (cell diameter 73.0-82.5 μm) and possesses robust denticles with broad blades and well-developed blade connections. Characterization of its primary and secondary SSU rDNA structures indicated that T. pseudoheterodentata was distinctly different from congeneric species in H12, H15, E10_1, and V4 regions. Phylogenetic analysis revealed that the genetic distances among the new species and similar species reached interspecific levels, furthermore, the phylogenetic study also validated the identification of T. pseudoheterodentata and its placement in the genus Trichodina.
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