Redescription and molecular characterization of two Trichodina species (Ciliophora, Peritrichia, Mobilida) from freshwater fish in China.

During an investigation of parasitic ciliates in northern China, two Trichodina species, T. acuta Lom, 1970 and T. nigra Lom, 1960, were isolated from the freshwater fish Cyprinus carpio Linnaeus, 1758. The morphology of each species was investigated based on dry silver nitrate-stained specimens. In addition, the molecular phylogeny of each was analyzed based on small subunit ribosomal DNA (SSU rDNA) sequence data. Trichodina acuta can be distinguished from its congeners by the undefined periphery of the central circle, the distinct gap between the rays and the central circle, and the distinctly sickle-shaped blades. Trichodina nigra is a cosmopolitan ciliate and is characterized by its densely linked denticles, broad, rounded spatula-shaped blades, robust central parts, and well developed rays. Phylogenetic analyses revealed that T. acuta and T. nigra nest within different clades, supporting the assertion that the GC content of SSU rDNA sequences could reflect evolutionary relationships among Trichodina species.

Morphology, morphogenesis and phylogeny of Anteglaucoma orientalis n. sp. (Ciliophora, Oligohymenophorea), a freshwater hymenostomatid from northeastern China.

A new hymenostomatid ciliate, Anteglaucoma orientalis n. sp., isolated from a freshwater pond in Harbin, northeastern China, was investigated using live observation and silver staining methods. Anteglaucoma orientalis is characterized as follows: size in vivo about 50-60 × 30-35 µm; oval body shape; buccal area occupies about 25% of body length; 28-36 somatic kineties; membranelle 1 having six or seven basal body rows, membranelle 2 five to seven rows, and membranelle 3 three rows; single macronucleus with one micronucleus attached. Morphogenesis of the genus Anteglaucoma is revealed for the first time. The main events during binary fission are as follows: morphogenesis begins with proliferation of kinetosomes in the middle part of postoral kinety 1, and kinetosomes of this primordial field multiply and organize to finally form the paroral membrane and membranelles 1-3 of the opisthe; the parental apparatus in the proter does not take part in the stomatogenetic process. Phylogenetic analyses based on SSU rRNA gene sequences show that Anteglaucoma orientalis n. sp. clusters with the type species, A. harbinensis Pan et al., 2017, with full support.

Morphology and molecular phylogeny of three Epistylis species found in freshwater habitats in China, including the description of E. foissneri n. sp. (Ciliophora, Peritrichia).

The peritrich genus Epistylis is speciose, however many species lack complete morphological description based on modern criteria and/or molecular data. In the present study, one new species, i.e., E. foissneri n. sp., and two morphologically similar species, i.e., E. hentscheli Kahl, 1935 and E. vaginula Stokes, 1884, collected from freshwater habitats in China, were studied. Epistylis foissneri n. sp. is characterized by its extremely slender zooids encased in a gelatinous sheath, symmetrically dichotomously branched stalk, trochal band located at mid-body, contractile vacuole located on dorsal wall of infundibulum, infundibular polykinety 3 (P3) composed of three equal-length rows that terminate above infundibular polykinety 1 (P1), 105-110 silverlines between the peristome and the trochal band, and about 110 silverlines between the trochal band and the scopula. Epistylis hentscheli is characterized by its asymmetric pyriform zooids (average length ca. 160 µm in vivo), dichotomously branched stalk with transverse striations on the surface of the upper portion, P3 three-rowed and terminating slightly above P1, 60-75 silverlines between the peristome and the trochal band, and 55-90 silverlines between the trochal band and the scopula. Epistylis vaginula is characterized by its elongated body shape (about 100 µm in length in vivo), dichotomously branched and smooth stalk, P3 three-rowed and terminating above P1, 80-100 silverlines between the peristome and the trochal band, and 45-80 silverlines between the trochal band and the scopula. The small subunit ribosomal DNA gene (SSU rDNA) of these three species was sequenced and supported the validity of each. Phylogenetic analyses based on SSU rDNA sequence data revealed that all three morphospecies group with other congeners within the major clade of Epistylis.

First record of dark leathery surface of geoduck clams in Panopea globosa.

Dark Leathery Surface of Geoduck Clams (LSGC) is an alteration that affects the periostracum of the mantle and siphon of Panopea generosa from the northwest coast of Canada and Mexico. This alteration affects commercialization and possibly the survival of the clams. The cause of LSGC is unknown but has been correlated with presence of fungi and protozoans. We detected a similar alteration in Panopea globosa from Baja California, Mexico and the histophagous ciliate Uronema marinum was isolated from affected siphon tissue. U. marinum was identified by its morphology and by genetic analysis of the gene 18S rRNA. This is the first record of LSGC in P. globosa and the first identification of a histophagous protozoan associated with it.

Morphological, Phylogenetic and Ecophysiological Characterization of a New Ciliate, Platynematum rossellomorai n. sp. (Oligohymenophorea, Scuticociliatia), Detected in a Hypersaline Pond on Mallorca, Spain.

With highly specialized morphology and unexplored functional capacities, ciliates from extreme habitats are drawing increasing attention. During a microbial investigation of a solar saltern pond (salinity 240‰) on Mallorca, Spain, a previously unknown scuticociliate, Platynematum rossellomorai n. sp. was isolated, cultured and studied using a tripartite approach consisting of a morphological description, a molecular analysis and an ecophysiological characterization. The ciliate has distinct morphological characteristics and its main diagnostic features include a large anteriorly positioned oral area (occupying almost half of the body length), two caudal cilia and a small number of somatic kineties. However, due to the most important generic feature of Cinetochilidae, the consistency of the arrangement of the adoral membranes, the ciliate is classified as a new member of the genus Platynematum. Its 18S rRNA gene sequence shows a sequence similarity of 91.0% to the closest deposited relative, Platynematum salinarum, and a phylogenetic analysis reveals a close relationship to other members of the family Cinetochilidae Perty, 1852. Growth experiments identify the ciliate as a borderline halophile, with a tolerance range between 180 and 280‰ salinity. The ciliate apparently accumulates the compatible solutes glycine betaine and ectoine to counterbalance osmotic stress, however, other osmoregulatory mechanisms are not excluded.

Morphology, molecular phylogeny, and taxonomy of trichodinids (Ciliophora, Mobilida) from Calanoid copepods.

Trichodinid ciliates were isolated from calanoid copepods and were characterized by morphological and molecular techniques. The trichodinids found in this study were morphologically similar to Trichodina acuta, but morphometric differences were observed between our specimens and T. acuta. The T. acuta specimens, isolated from fish in several geographical locations around the world, were slightly larger than specimens in our population. In fact, our morphological and morphometric data are more similar to Trichodina diaptomi populations, also found on copepods. Moreover, our phylogenetic analyses revealed that the 18S rRNA gene sequence from our samples emerged in a clade with two T. acuta sequences, infesting fish hosts in China. These three sequences showed 100% identity. Our data, along with a taxonomic review, suggest that the species T. diaptomi, described recently, is actually a synonym of T. acuta and that this species may be related to fish and copepod hosts. In conclusion, we highlight the urgent need to better investigate the polymorphism and host specificity within the Mobilida, as this data may represent important characters to better understand the evolution into this order.

Taxonomy and molecular phylogeny of three freshwater scuticociliates, with descriptions of one new genus and two new species (Protista, Ciliophora, Oligohymenophorea).

Three freshwater scuticociliates, Apouronema harbinensis gen. nov. spec. nov., Cyclidium vorax spec. nov., and C. glaucomaMüller, 1773, collected from rivers in Hulan District, Harbin, northeastern China, were investigated using morphological and phylogenetic criteria. Apouronema gen. nov., assigned to the family Uronematidae, is mainly distinguished from the other genera of the family by its paroral membrane extending anteriorly to the middle of membranelle 1. Apouronema harbinensis spec. nov. is defined by body size in vivo about 45-55 × 20-25 µm, buccal field about 70-80% of cell length; 12 or 13 somatic kineties; membranelle 1 having two rows, with 16-18 basal bodies in each kinety; membranelle 2 and membranelle 3 both having two rows each; scutica X-shaped with five pairs of basal bodies. Cyclidium vorax spec. nov. is characterized by the following features: body size 35-40 × 18-20 µm in vivo; 9 or 10 somatic kineties; membranelle 1 having two longitudinal rows, much shorter than M2; M2 triangle-shaped. The phylogenetic analyses show that: (1) Apouronema clustered in the Uronematidae clade, and grouped with genera Uronemita and Uronema; (2) Cyclidium vorax spec. nov. grouped with C. glaucoma and C. sinicum, which supports the assignment of the new species to the genus Cyclidium; (3) Cyclidium remains non-monophyletic with the addition of the new sequence.

Hyalophysa lynni n. sp. (Ciliophora, Apostomatida), a new pathogenic ciliate and causative agent of shrimp black gill in penaeid shrimp.

The parasitic ciliate causing shrimp black gill (sBG) infections in penaeid shrimp has been identified. The sBG ciliate has a unique life cycle that includes an encysted divisional stage on the host's gills. The ciliature of the encysted trophont stage has been determined and is quite similar to that of the closely related apostomes Hyalophysa bradburyae and H. chattoni. Hyalophysa bradburyae is a commensal ciliate associated with freshwater caridean shrimp and crayfish, while H. chattoni is a common commensal found on North American marine decapods. Based on 18S rRNA gene sequence comparisons, the sBG ciliate is more closely related to the marine species H. chattoni than to the freshwater species H. bradburyae. The unique life cycle, morphology, 18S rRNA gene sequence, hosts, location, and pathology of the sBG ciliate distinguish this organism as a new species, Hyalophysa lynni n. sp.

Morphology and molecular analyses of four epibiotic peritrichs on crustacean and polychaete hosts, including descriptions of two new species (Ciliophora, Peritrichia).

Four epibiotic sessilid peritrichs, i.e., Zoothamnium wilberti n. sp., Baikalonis microdiscus n. sp., Epistylis anastatica (Linnaeus, 1767) Ehrenberg, 1830, and Rhabdostyla commensalisMöbius, 1888, were isolated from one syllid polychaete and three crustacean hosts in Qingdao, China. For each species, specimens were observed both in vivo and following silver staining. Their SSU rDNA was also sequenced for phylogenetic analyses. Zoothamnium wilberti n. sp. is characterized by the appearance of its colony, which is up to 350 µm high, and usually has fewer than 16 zooids, and the dichotomously branched stalk with transverse wrinkles, the conspicuously conical peristomial disc, and infundibular polykinety 3 comprising three isometric ciliary rows. Baikalonis microdiscus n. sp. can be recognized by its barrel-shaped zooid, small peristomial disc, smooth and short stalk, and its unusual infundibular polykinety 3 comprising a long inner row and a short outer row. Two poorly known species, i.e., Epistylis anastatica and Rhabdostyla commensalis, are redescribed and redefined. Phylogenetic analyses reveal that: (i) R. commensalis is closely related to the family Astylozoidae rather than to the morphologically similar Epistylididae; (ii) B. microdiscus n. sp. is sister to the family Scyphidiidae; (iii) E. anastatica groups with vorticellids and ophrydiids, which further supports the polyphyly of the genus Epistylis; and (iv) Z. wilberti n. sp. is nested within the Zoothamniidae, as expected.

The complete life cycle of the unusual apostome Hyalophysa clampi (Ciliophora, Apostomatida), a symbiont of crayfish in Alabama (USA).

Hyalophysa clampi Browning and Landers, 2012 was reexamined to determine all stages in the life cycle of this symbiotic ciliate. The cell feeds as a normal exuviotroph within the exoskeleton of its molted crayfish host but does not encyst following the trophont stage. Trophonts transform into swimming tomont stages, which divide by palintomy over successive divisions, splitting to two cells, separating, and repeating. The divisions cease when the daughter cells attain the size of the infestive tomite stage, which attaches to a new crayfish. This unique life cycle is most similar to the European hermit crab symbiont Polyspira delagei, which forms chains of daughter cells during division. Scanning electron microscopy confirmed the unusual presence of two contractile vacuoles in H. clampi, unique among the Apostomatida, and provided ultrastructural details to better understand light microscopy silver staining. The genus diagnosis for Hyalophysa is modified herein to accommodate this new life cycle.

Morphological and molecular characterization of two Trichodina (Ciliophora, Peritrichia) species from freshwater fishes in China.

In the present study, we provide morphological and molecular characterization of two Trichodina species, T. acuta Lom, 1970 and T. funduliWellborn, 1967, isolated from koi (Cyprinus carpio) and loach (Paramisgurnus dabryanus), respectively. Morphological characters of the two Trichodina species were mainly investigated on the basis of dry silver nitrate-impregnated specimens. Both species are medium-sized and possess well-developed denticles comprising strongly sickle-shaped blades, well-developed central parts, and straight rays. Trichodina acuta can be easily distinguished from the other Trichodina species that possess a clear central circle by the well-developed sharp blade apophysis, and the gap between ray tip and central circle. Trichodina funduli is a poorly known species that is easily confused with T. heterodentata Duncan, 1977, however the latter species has thinner denticles. The small subunit ribosomal RNA gene sequences of Trichodina acuta and T. funduli were incorporated into phylogenetic analyses. Our findings suggest that the phylogenetic lineage of trichodinids might not correspond with their living environments, host species or even some morphological characteristics.

Taxonomy and phylogeny of Pseudovorticella littoralis sp. n. and P. alani sp. n. (Ciliophora: Peritrichia) from coastal waters of southern China.

The morphology, infraciliature, and silverline system of two peritrich ciliates, Pseudovorticella littoralis sp. n. and P. alani sp. n., isolated from coastal waters of southern China, were investigated based on both living and silver-stained specimens. Pseudovorticella littoralis sp. n. is characterized by the following combination of characters: cell inverted cone-shaped; contractile vacuole ventral; J-shaped macronucleus; infundibular polykinety 3 with two kinetosome rows of equal length; 19-26 silverlines from peristome to trochal band and 5-14 from trochal band to scopula. Pseudovorticella alani sp. n. is characterized by: cell inverted bell-shaped; contractile vacuole ventral; J-shaped macronucleus recurved almost forming a loop; infundibular polykinety 3 with three kinetosome rows, outer two rows longer than inner one; 48-61 silverlines between peristome and aboral trochal band, and 12-20 between aboral trochal band and scopula. The SSU rDNA sequences of both new species are reported and their genetic distances with congeners and phylogenetic relationships are investigated. Pseudovorticella and Epicarchesium cluster into two subclades with low support values. One subclade contains nearly all the available sequences of Pseudovorticella and Epicarchesium. Another one contains P. monilata and E. pectinatum. This calls on the need of a generic re-classification of Pseudovorticella and Epicarchesium based on more morphological and molecular data.

Morphology and Phylogeny of Three Pseudovorticella Species (Ciliophora: Peritrichia) from Brackish Waters of China.

The biodiversity of peritrich ciliates from brackish biotopes is rarely investigated, especially members of the genus Pseudovorticella. Here, the morphology of three species of Pseudovorticella, i.e. P. cf. vestita (Stokes, 1883) Jankowski, 1976, P. spathulata sp. n., and P. qinghaiensis sp. n. isolated from brackish waters were studied. Pseudovorticella cf. vestita is characterized by inverted bell-shaped cell; a J-shaped macronucleus; a single contractile vacuole ventrally located; P3 three-rowed; pellicle striated with highly developed pellicular vesicles; 18-22 transverse silverlines between peristome and aboral trochal band, and 9-13 between aboral trochal band and scopula. Pseudovorticella spathulata sp. n. differs from its congeners by the following combination of characters: elongate-elliptical cell; a single contractile vacuole near ventral wall of infundibulum; a J-shaped macronucleus; P3 three-rowed; 24-34 silverlines between oral area and aboral trochal band and 6-10 between aboral trochal band and scopula. Pseudovorticella qinghaiensis sp. n. is characterized by: cell with an oval outline; a single contractile vacuole near ventral wall of infundibulum; a C-shaped macronucleus; P3 three-rowed; 30-35 and 9-11 transverse silverlines above and below the trochal band, respectively. The SSU rDNA sequences of five Pseudovorticella species, namely P. annulata, P. monilata, P. parakenti, P. spathulata sp. n., and P. cf. vestita, plus that of Zoothamnium hartwigi, are reported for the first time and their evolutionary relationships are investigated. Five undefined Pseudovorticella forms are considered might be conspecific with P. monilata. Two congeners are conspecific with P. spathulata sp. n. Phylogenetic analyses based on SSU rDNA sequences reveal that Pseudovorticella is not monophyletic and Z. hartwigi clusters with its congeners as expected.

Morphological and Molecular Identification of a New Ciliate, Zoothamnium palmphlatum nov. spec. (Ciliophora, Peritrichia) from North China.

A new peritrichous ciliate, Zoothamnium palmphlatum nov. spec., was collected from an estuary in Yantai, China. It was investigated, using both live observation and silver staining. The new species can be identified by a palm-shaped colony consisting of highly developed and alternately arranged secondary branches, a double-layered peristomial lip, and an infundibular polykinety 3 composed of two parallel kinetosomal rows. Phylogenetic analyses of the small subunit (SSU) rRNA gene sequence show that Z. palmphlatum clusters with other members of the family Zoothamniidae. Furthermore, the comparison of primary and secondary SSU rDNA structures indicates that Z. palmphlatum is distinctly different from its morphologically similar species (93.2-97.0% in sequence similarity) in combination of H10 and H31 regions. Although Z. palmphlatum shares highest sequence similarity with Zoothamnium mucedo (98.9%), the new species has distinctly different structures in the H11, H12, H18, and H31 regions compared to Z. mucedo, which indicates that sequence similarity may not determine the similarity of the secondary structure.

Morphological Redescription and Molecular Identification of Trichodina reticulata Hirschmann & Partsch, 1955 (Ciliophora, Mobilida, Trichodinidae) with the Supplemental New Data of SSU rDNA and ITS-5.8S rDNA.

During a survey of trichodinids in Chongqing, China, two populations of Trichodina reticulata Hirschmann & Partsch, 1955 were isolated from gills of the freshwater fishes, Carassius auratus and Ctenopharyngodon idellus, and 11 molecular samples containing small subunit (SSU) rDNA and internal transcribed spacer (ITS)-5.8S rDNA were newly sequenced. Both populations fell within the range of morphometry and corresponded well with the diagnostic morphological features of Trichodina reticulata Hirschmann & Partsch, 1955. It should be noted that one population possessed obvious central granules in the adhesive disc, while the other one did not. The detailed morphological redescription for these two populations is given in this work. Although some intraspecific differences were found, both populations were confirmed as Trichodina reticulata based on the morphological study and molecular data, including the sequence analysis of the primary and secondary structures of SSU rDNA, genetic distances and phylogenetic tree. Our study supports the assertion that central granules in the adhesive disc might be an auxiliary feature rather than a heritable character for determining taxonomic affiliations among trichodinids.

Morphology and Phylogeny of Four New Vorticella Species (Ciliophora: Peritrichia) from Coastal Waters of Southern China.

Four new species of Vorticella, V. parachiangi sp. n., V. scapiformis sp. n., V. sphaeroidalis sp. n., and V. paralima sp. n., were isolated from coastal brackish waters of southern China. Their morphology, infraciliature, and silverline system were investigated based on observations of specimens both in vivo and following silver staining. Vorticella parachiangi sp. n. is distinguished by: a J-shaped macronucleus; a single dorsally located contractile vacuole; a two-rowed infundibular polykinetid 3, in which row 1 is shorter than row 2; 21-31 silverlines between peristome and aboral trochal band, 6-11 between aboral trochal band and scopula. Vorticella scapiformis sp. n. is characterized by its conspicuously thin and irregularly edged peristomial lip; a J-shaped macronucleus; a single, ventrally located contractile vacuole; row 1 of the infundibular polykinetid 3 proximally shortened; 18-25 silverlines between peristome and aboral trochal band, 8-12 between aboral trochal band and scopula. Vorticella sphaeroidalis sp. n. can be identified by its small, sub-spherical zooid; a C-shaped macronucleus; a ventrally located contractile vacuole; an aboral trochal band adjacent to the scopula; 16-18 silverlines between persitome and aboral trochal band, two between aboral trochal band and scopula. Vorticella paralima sp. n. can be identified by its ovoidal zooid; a J-shaped macronucleus; a dorsally positioned contractile vacuole; rows 1 and 2 of the infundibular polykinetid 3 proximally shortened; 26-35 silverlines from peristome to aboral trochal band, and 7-13 from aboral trochal band to scopula. The SSU rDNA genes of these four species were sequenced and their phylogeny was analyzed.

Trichodina amblypharyngodoni sp. n. and Trichodina hoffmani Wellborn, 1967 (Ciliophora: Trichodinidae) from the freshwater fishes in the Baikka Beel of Moulvibazar district in Sylhet division, Bangladesh

During a survey on species diversity of trichodinid ciliates from freshwater fishes in the Baikka Beel of Moulvibazar district in Sylhet division from January to December 2015, two species of ciliates were identified, of which one is new to science. Trichodina hoffmani Wellborn, 1967 is collected from the gills of Mystus tengara, and Trichodina amblypharyngodoni sp. n. is described from Amblypharyngodon mola using the silver nitrate impregnation method. T. amblypharyngodoni sp. n. is characterized by having a large, darkly stained central area; rectangularly-rounded butterspoon-shaped blade with very narrow interblade space and blunt tangent point; indistinct anterior and posterior blade apophysis, but a rounded apex near the base of the central part of denticle; slender, sometimes slightly triangular central part with rounded tip; and filamentous, curved, anteriorly directed rays with inflated tip and indistinct central groove. Based on these characters and the unique shape and absence of variability of the denticles among the silver impregnated specimens of the present species, it may be said that to a lesser extent, it resembles Trichodina prowazeki Grupcheva and Lom, 1980.

Dawn of astome ciliates in light of morphology and time-calibrated phylogeny of Haptophrya planariarum, an obligate endosymbiont of freshwater turbellarians.

Morphology, systematic position and time-calibrated phylogeny of Haptophrya planariarum were investigated. This endosymbiont of freshwater turbellarians is characterized by: (i) a length of about 200-900 µm; (ii) a campanulate to truncate claviform body carrying an anterior adhesive sucker; (iii) an ellipsoidal macronucleus localized in the rear body end; (iv) a contractile canal extending along the dorsal margin; and (v) usually more than 150 meridional ciliary rows, a horseshoe-shaped suture line along the sucker, and two inconspicuous secant systems at lateral ends of the suture line. In 18S rRNA gene phylogenies, astomes were depicted as a non-monophyletic group within the scuticociliate clade, whereby H. planariarum clustered with the loxocephalid genus Dexiotricha. After considering morphological evidence, statistical tree topology tests and evolutionary distances, we find astomes as a distinct group that evolved from a free-living scuticociliate ancestor in the early Paleozoic. Molecular clock analyses indicated that astomes living in annelids diverged from those inhabiting turbellarians within about 50 Ma during the Late Cambrian and the Upper Ordovician. This comparatively short time span might have not sufficed for fixation of molecular synapomorphies in the 18S rRNA gene and/or they might have been erased by substitutions during the almost 500 Ma-long evolutionary history of astomes.

A new species of Trichodina Ehrenberg, 1830 (Ciliophora: Trichodinidae) infecting farmed Clarias gariepinus (Burchell) (Siluriformes: Clariidae) in Cuba.

A new species of Trichodina Ehrenberg, 1830 collected from the skin and fins of farmed North African catfish Clarias gariepinus (Burchell) fingerlings, is described. The new species can be distinguished from other trichodinids by the characteristics of the adhesive disc, especially by the great number of denticles. Trichodina merciae n. sp. is morphologically similar to T. renicola (Mueller, 1931) and T. marplatensis Martorelli, Marcotegui & Alda, 2008, in the number of denticles, but differs in the morphometric data, denticle morphology, environment and location. Trichodina merciae n. sp. has broad sickle-shaped blades and thin, straight rays, while T. marplatensis has broad club-shaped blades and wide S-shaped rays. Besides, denticle length, blade length, ray length, width of central part and denticle span of the new species are greater than T. marplatensis. However, the diameter of denticle ring and the diameter of the central area in T. marplatensis is larger than the ones in T. merciae n. sp. This is the first record of freshwater ectoparasite trichodinid with an average number of denticles greater than 50.

New data of two trichodinid ectoparasites (Ciliophora: Trichodinidae) from farmed freshwater fishes in Hubei, China.

Trichodinids are ciliated protozoan parasites or symbionts of marine and freshwater organisms. During a parasitic ciliate survey in Hubei, China, two Trichodina species, T. acutaLom, 1961 and T. hyperparasitis Chen & Hsish, 1984, were isolated from body surface of loach Misgurnus anguillicaudatus and gills of yellow catfish Pelteobagrus fulvidraco, respectively. The two species were investigated morphologically, based on silver-strained specimens, and their small subunit ribosomal RNA gene (SSU rDNA) sequences were sequenced. Morphologically, T. acuta is distinguished mainly by the acute sickle-shaped blade with well-developed sharp blade apophysis, the irregular circular granule, and the robust ray. Trichodina hyperparasitis is mainly characterized by the broad blade and the inconspicuous round ray apophysis. Phylogenetic analyses revealed that the two Trichodina species investigated were nested within a clade including several freshwater Trichodina species. This indicates that the central granule is a useful taxonomic feature, but it may not be an important phylogenetic characteristic. Histopathological analysis revealed that T. hyperparasitis firmly colonized gills, which resulted in discrete hyperplasia and injuries of the gill filaments.