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A new ciliate species of the genus Loxophyllum Dujardin, 1841, Loxophyllum paludosum sp. n., is described from a mangrove wetland near Daya Bay in Guangdong Province, southern China, based on morphological and molecular analyses. The new species can be distinguished from its congeners by a combination of the following characters: (1) 12-14 right kineties and 4-6 left kineties; (2) two macronuclear nodules and one micronucleus; (3) a single contractile vacuole located terminally; (4) extrusomes bar-shaped, evenly spaced along entire ventral margin, and clustered to form 5-7 warts along dorsal margin; and (5) presence of three ridges on the left side of cell. The new species is divergent from its congeners from 0.4% to 6.7% (5-104 nucleotide sites) based on the small subunit (SSU) rRNA gene sequence data. The validity of the new species is also supported by molecular phylogenetic trees inferred from SSU rRNA gene sequences., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

The morphology of two new scuticociliates, Uronema apomarinum sp. nov. and Homalogastra parasetosa sp. nov., isolated from a mangrove wetland in Shenzhen, PR China, was studied using live observation and the protargol impregnation method. Uronema apomarinum is characterized by a body size of about 20-35×10-15 µm in vivo , a partly two-rowed membranelle 1, and 12 or 13 somatic kineties. Homalogastra parasetosa is distinguished by a membranelle 1 comprising two longitudinal rows of basal bodies. Three Homalogastra setosa populations are suggested as subjective synonyms of the new species. Improved diagnoses of the genus Homalogastra Kahl, 1926 and its type species Homalogastra setosa Kahl, 1926 are provided. Results of phylogenetic analyses based on 18S rRNA gene and ITS1-5.8S-ITS2 region sequences indicate that U. apomarinum is most closely related to U. marinum , while the closest relative of H. parasetosa is H. setosa .

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., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

The morphology, morphogenesis and molecular phylogeny of the oxytrichid ciliate, Monomicrocaryon euglenivorum euglenivorum (Kahl, 1932) Foissner, 2016, isolated from freshwater in a seaside park, Guangzhou, China, were investigated. Monomicrocaryon euglenivorum euglenivorum can be recognized as follows: caudal cirri in midline of body; dorsal kinety 1 without a one-kinetid-wide gap; transverse cirri acicular or rod-shaped with a fringed distal end; right marginal row commences at level of buccal vertex or anterior to buccal vertex. The main events during binary fission are as follows: (1) the proter retains the parental adoral zone of membranelles entirely; (2) frontoventral-transverse cirral anlagen I-VI are segmented in the ordinary pattern 1:3:3:3:4:4 from left to right, which form three frontal, four frontoventral, one buccal, three postoral ventral, two pretransverse ventral and five transverse cirri, respectively; (3) dorsal morphogenesis is in the typical Oxytricha-pattern, but fragmentation of dorsal kinety 3 is indistinct; and (4) three caudal cirri are formed, one at the posterior end of each of dorsal kineties 1, 2 and 4. Phylogenetic analyses based on SSU rDNA sequences showed that M. euglenivorum euglenivorum clustered with Kleinstyla dorsicirrata and Heterourosomoida lanceolata rather than with its congener M. elegans. The genus Monomicrocaryon is not monophyletic in this study; however, its monophyly is not rejected by the AU test., (Crown Copyright © 2019. Published by Elsevier GmbH. All rights reserved.)

Questions regarding ciliate distribution (endemism vs. cosmopolitanism) and degree of genetic diversity (high vs. low) remain unsettled, even when the same organism is under investigation. Presence of genes with high copy number and amplification of non-dominant haplotypes might account for the observed discordance in these studies. Herein, we used direct PCR and cloning sequencing to examine intra-population sequence diversity and its effect on assessments of phylogeography of Paramecium multimicronucleatum. Totally, 381 ITS1-5.8S rDNA-ITS2-28S rDNA and 304 mitochondrial cytochrome oxidase subunit I (COI) gene sequences were generated for 18 populations of P. multimicronucleatum. The following results were obtained: (1) Direct sequencing of PCR products captured the dominant ITS and LSU haplotypes, indicating that it is an appropriate strategy for constructing phylogeography of large-scale spatial populations. (2) Deep cloning was deemed more appropriate for the COI gene for population level studies, as direct sequencing could not easily capture the dominant haplotypes. (3) No endemic populations of P. multinucleatum were noted, indicating origin from a single founder population. (4) Nuclear genetic diversity within temporal populations was high, but only the dominant haplotypes seemed to be passed on to subsequent generations., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

Three new cyrtophorian ciliates isolated from coastal areas of China were described based on morphological and genetic data. The Chlamydodon mnemosyne-like species Chlamydodon similis sp. n. differs from its congeners mainly by its number of somatic kineties. Chlamydodon oligochaetus sp. n. is distinguished from its congeners mainly by having fewer somatic kineties, and/or an elongated body shape. Chlamydodon crassidens sp. n. is characterized mainly by an inverted triangular body shape, a posteriorly interrupted cross-striated band (5-6 μm wide), and a large cytostome. Moreover, we provided small-subunit (SSU) rDNA sequences of C. similis sp. n. and C. oligochaetus sp. n. Maximum likelihood (ML) and Bayesian inference (BI) consistently placed C. similis sp. n. as a sister to C. paramnemosyne, but showed different branching position of C. oligochaetus sp. n., which may be due to a low taxon sampling in the Chlamydodontidae and/or an insufficient resolution of the marker gene at species level., (© 2017 The Author(s) Journal of Eukaryotic Microbiology © 2017 International Society of Protistologists.)

Two oxytrichid ciliates, Stylonychia (Metastylonychia) nodulinucleata Shi and Li, 1993 and Gastrostyla steinii Engelmann, 1862; collected from Huguangyan Maar Lake and Shenzhen Mangrove Nature Protection Area, southern China, respectively, were investigated using standard methods. The uncommon species Stylonychia (Metastylonychia) nodulinucleata can be recognized by its large body size, the conspicuous caudal cirri in vivo, and a moniliform macronucleus. We provide the first record of the small subunit ribosomal RNA (SSU rRNA) gene sequence for the species. The division of Stylonychia into two subgenera based on morphological and morphogenetic data is supported by the phylogenetic analyses, in which Stylonychia (Metastylonychia) nodulinucleata is placed as a sister branch to the clade including all the sequences of Stylonychia (Stylonychia) mytilus complex. The first brackish water population of Gastrostyla steinii is described in detail, with emphasis on its morphogenesis, which corresponds well with previous populations. In the phylogenetic trees, all the available Gastrostyla sequences, except for Gastrostyla sp. Y2 (KT780432) (probably a misidentification), nest together in the big group of the subfamily Stylonychinae with moderate to high support (ML/BI, 91%/0.95), very likely revealing the monophyly of the genus Gastrostyla., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

The morphology, infraciliature, and silverline system of three peritrichous ciliates, Zoothamnium bucciniiformum sp. n., Zoothamnium florens sp. n., and Zoothamnium zhanjiangense sp. n., were investigated based on both living and silver-stained specimens. Zoothamnium bucciniiformum sp. n., collected from coastal waters (salinity 30‰) off Zhanjiang, southern China, can be distinguished by the following characters: dichotomously branched stalk, peristomial lip with medial circumferential infolding, contractile vacuole apically positioned, 32-49 silverlines between the anterior end and the aboral trochal band, 15-26 between the aboral trochal band and the scopula; two kineties in peniculus 3, not parallel to each other. Zoothamnium florens sp. n., collected from a mangrove wetland (salinity 13‰) off Zhanjiang, is characterized by its large conical zooid, tuberculate peristomial lip, asymmetrical dichotomously branched colony, 59-81 silverlines between the anterior end and the aboral trochal band and 29-36 between the aboral trochal band and the scopula. Zoothamnium zhanjiangense, collected from a mangrove wetland (salinity about 9.5‰) off Zhanjiang, differs from its congeners by the alternately branched stalk, peristomial lip with medial circumferential infolding, 40-63 silverlines from the peristomial area to the aboral trochal band and 13-24 from the aboral trochal band to the scopula. The comparison and analysis of SSU rDNA sequences also support present identifications., (© 2016 The Author(s) Journal of Eukaryotic Microbiology © 2016 International Society of Protistologists.)

The ciliate genus Protocruzia is a highly confused group, which was formerly placed in the class Heterotrichea or Karyorelictea, and is according to the most recent system tentatively assigned to the class Spirotrichea. In the present study, the morphology, ciliary pattern, and molecular phylogeny of two poorly known species, Protocruzia tuzeti Villeneuve-Brachon, 1940, and Protocruzia granulosa Kahl, 1933, isolated from coastal waters of China, were investigated. Protocruzia tuzeti differs from its congeners mainly in possessing 6 adoral membranelles, 8-11 somatic kineties, and postoral dikinetids. Protocruzia granulosa is characterized by its extremely slender body, three postoral kineties, and 13 or 14 somatic kineties. The morphogenesis of P. granulosa is similar to that of P. tuzeti, especially in the parakinetal mode of stomatogenesis and the reorganization of the parental paroral membrane; however, more than one somatic kinety joins in the formation of the oral primordium in P. granulosa. Phylogenetic analyses based on small subunit ribosomal RNA gene revealed that six Protocruzia species form a fully supported clade that does not belong to any ciliate class; therefore, our data support the establishment of the class Protocruziea Gao et al. (Sci. Rep., 6, 2016, 24874)., (© 2016 The Author(s) Journal of Eukaryotic Microbiology © 2016 International Society of Protistologists.)

The morphology, ciliary pattern and small subunit rDNA (SSU rDNA) sequences of two new cyrtophorian ciliates, Aegyria foissneri sp. n. and Lynchella minuta sp. n., isolated from brackish waters in southern China, were investigated. Aegyria foissneri sp. n. is characterized as follows: cell size 85-170×45-80μm in vivo; body inverted oval with a protrusion and a dark pigment spot on anterior left part; 42-77 somatic kineties; one preoral and three to six circumoral kineties; five to eight transpodial segments; 31-44 nematodesmal rods; 12-16 contractile vacuoles; and single oval macronucleus. Lynchella minuta sp. n. is distinguished from its congeners by having a cell size of 20-30×15-20μm in vivo, oval body outline; four preoral and 14 or 15 postoral kineties, three circumoral kineties; ca. 11 nematodesmal rods; one finger-like tentacle on the ventral side; and two diagonally located contractile vacuoles. Molecular phylogenetic analyses support the genus assignment of Aegyria foissneri sp. n. and indicate the monophyly of the genus. While Lynchella minuta sp. n. clusters with Coeloperix species, which indicates that Lynchella is non-monophyletic., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Haptorian ciliates in the closely similar genera Kentrophyllum and Epiphyllum possess a unique pattern of ciliature and are distinguished from one another only by the presence or absence of cytoplasmic spines projecting from the margin of the cell. Phylogenetic analyses based on SSU rDNA sequences of three new samples from coastal habitats in China revealed that species in the two genera clustered together indiscriminately (i.e. forms of neither genus clustered into an independent clade) as a maximally supported, monophyletic clade that branches basally to all other clades in the order Pleurostomatida and is strongly divergent from other members of the family in which the genera have been placed. As a result, we propose that Epiphyllum be synonymized with Kentrophyllum and that a new family Kentrophyllidae fam. n. be established for the genus. We hypothesize that the two-sutures of Kentrophyllum is a plesiomorphy within the Pleurostomatida and the unique peripheral kinety might represent an autapomorphy of Kentrophyllum. In addition, we provide a taxonomic revision of Kentrophyllum including description of three new species (K. bispinum sp. n., K. strumosum sp. n., and K. qingdaoense sp. n.), redescription of K. verrucosum (Stokes, 1893) Petz et al., 1995, and three new combinations (K. soliforme (Fauré-Fremiet, 1908) comb. n., K. hohuensis (Wang and Nie, 1933) comb. n. and K. shenzhenense (Pan et al., 2010) comb. n.). The surface ultrastructure of the genus Kentrophyllum is recorded for the first time. And a key to all known species of Kentrophyllum was also suggested.

The morphology and morphogenesis of an oxytrichid ciliate, Rubrioxytricha haematoplasma (Blatterer & Foissner, 1990) Berger, 1999, collected from brackish and marine waters in China, were investigated using live observation and the protargol staining method. The main features of the morphogenetic process are: (i) the parental adoral zone of membranelles is retained completely in the proter and the anlage of undulating membranes originates from dedifferentiation of the old structures; (ii) three frontal, four frontoventral, one buccal, five ventral and five transverse cirri are derived from the anlagen of the undulating membranes and the five streaks of frontal-ventral-transverse anlagen in the pattern of 1:3:3:3:4:4 from left to right; (iii) the morphogenesis of the dorsal kineties is simpler than the Oxytricha pattern, i.e. without fragmentation of the dorsal kinety 3 anlagen; (iv) the single caudal cirrus originates from the dorsal kinety 3 anlage on the right side; (v) the two macronuclear nodules fuse into a single mass during the mid-stage of morphogenesis. These features correspond well with Rubrioxytricha indica, indicating that the morphogenetic pattern of Rubrioxytricha is stable. Phylogenetic analysis based on small-subunit rRNA gene sequence data supports the monophyly of the genus Rubrioxytricha, which is nested within the non-Stylonychinae clade., (© 2015 IUMS.)

Two novel brackish water urostyloid ciliates, Anteholosticha paramanca sp. n. and Antiokeronopsis flava gen. n., sp. n., isolated from the Shenzhen Mangrove Nature Protection Area on the coast of the South China Sea, were investigated using live observation and protargol impregnation techniques. Anteholosticha paramanca sp. n. is characterized by its spherical yellowish cortical granules arranged in lines, shortened midventral complex and three transverse cirri. Morphogenesis is similar to that in Anteholosticha manca. The new genus Antiokeronopsis is diagnosed by having a continuous adoral zone of membranelles, frontal cirri arranged in a bicorona, midventral complex composed of midventral pairs only, one marginal cirral row on each side, the presence of frontoterminal and transverse cirri, and the lack of buccal and caudal cirri. The type species A. flava sp. n. is characterized by its elongated body shape, brown to yellowish body color and two types of cortical granules. Small subunit ribosomal RNA gene sequence data justify the classification of both species. Phylogenetic analyses indicate that A. paramanca clusters with Bakuella subtropica within a clade that includes two other Anteholosticha species, while Antiokeronopsis groups within the core urostylids and is most closely related to the well-known genera Pseudokeronopsis and Uroleptopsis., (© 2014 The Author(s) Journal of Eukaryotic Microbiology © 2014 International Society of Protistologists.)

In order to assess how dataset-selection for multi-gene analyses affects the accuracy of inferred phylogenetic trees in ciliates, we chose five genes and the genus Paramecium, one of the most widely used model protist genera, and compared tree topologies of the single- and multi-gene analyses. Our empirical study shows that: (1) Using multiple genes improves phylogenetic accuracy, even when their one-gene topologies are in conflict with each other. (2) The impact of missing data on phylogenetic accuracy is ambiguous: resolution power and topological similarity, but not number of represented taxa, are the most important criteria of a dataset for inclusion in concatenated analyses. (3) As an example, we tested the three classification models of the genus Paramecium with a multi-gene based approach, and only the monophyly of the subgenus Paramecium is supported., (Copyright © 2013 Elsevier Inc. All rights reserved.)

This paper investigates the morphology, morphogenesis and SSU rRNA gene-based phylogeny of Apoholosticha sinica n. g., n. sp., isolated from mangrove wetland in Shenzhen, southern China. The new genus Apoholosticha is characterized by its bipartite adoral zone, clearly differentiated frontal cirri arranged in a bicorona, midventral complex composed of midventral pairs only, one marginal cirral row on each side, presence of frontoterminal and transverse cirri, and the lack of a buccal cirrus and caudal cirri. The type species, Apoholosticha sinica n. sp. is diagnosed by the elongated body shape and two kinds of cortical granules. Its main morphogenetic features are similar to that of Pseudokeronopsis except for (1) no buccal cirrus is formed and (2) its macronuclear nodules fuse into a single mass during cell division. Phylogenetic analyses for the new taxon indicate that Apoholosticha n. g. is most closely related to Nothoholosticha and Heterokeronopsis, and falls into the family Pseudokeronopsidae within the core Urostylida clade. In addition, a species that had been misidentified in previous literature is here recognized and assigned to the new genus as Apoholosticha sepetibensis (Wanick and Silva-Neto, 2004) n. comb. (basionym: Pseudokeronopsis sepetibensis Wanick and Silva-Neto, 2004)., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

Oligotrich ciliates are common members of marine microplankton. However, their biodiversity is not well documented. In this study, the morphology and phylogenetic positions of three new oligotrich species, Spirostrombidium apourceolare spec. nov., Spirostrombidium subtropicum spec. nov. and Parallelostrombidium conicum spec. nov., collected from coastal habitats of southern China, were investigated. Spirostrombidium apourceolare is characterized by the girdle kinety which encircles the cell twice as two dextrally oriented whorls with some undulations and by the presence of several macronuclear nodules. Spirostrombidium subtropicum is recognized by the girdle kinety encircling the cell as two dextrally oriented whorls and extrusomes arranged in a stripe along the girdle kinety. Parallelostrombidium conicum differs from its congeners by the obconic body shape and the posterior portion of the girdle kinety extending downwards on the left ventral side to reach the posterior pole. In small-subunit rRNA gene trees, S. subtropicum clusters with Omegastrombidium elegans and Varistrombidium kielum, and Parallelostrombidium conicum is sister to the clade containing Novistrombidium sinicum, Novistrombidium orientale and Parallelostrombidium sp.

This paper investigates the morphology, ontogenesis and small subunit (SSU) rRNA gene-based phylogeny of a new urostylid ciliate, Bakuella subtropica sp. n., discovered from the estuary of the Pearl River in Guangzhou, southern China. The new species is diagnosed by its elongate body, one buccal and one parabuccal cirrus, midventral complex comprised of 9-23 midventral pairs and one or two midventral rows extending to four fifths of body length, yellow-brown to yellow-greenish cortical granules and an estuary habitat. Its main ontogenetic features are: (1) in the proter, the parental adoral zone of membranelles is completely renewed by new structures and old midventral pairs join the formation of frontal-midventral-transverse cirral anlagen (FVT-anlagen); (2) in the opisthe, the oral primordium originates apokinetally, FVT-anlagen are formed besides and some old midventral cirri join the formation; (3) the anlagen for marginal rows and dorsal kineties develop intrakinetally; and (4) the numerous macronuclear nodules fuse into a single mass before dividing. Based on the SSU rDNA sequences, phylogenetic analyses show a close relationship between Bakuella subtropica sp. n., Apobakuella and Neobakuella, forming a clade separated from the other genera in the family Bakuellidae. Available morphological and ontogenetic data challenge the monophyly of Bakuellidae., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

Two new species of the taxonomically confused genus Amphisiella were isolated from brackish-water habitats in southern China, and their morphology and morphogenesis were investigated. The genes coding for small subunit (SSU) rRNA were sequenced in each species and included in a phylogenetic analysis with all data available from nominal species of Amphisiella and related hypotrichs. Both new species are diagnosed by an elongate body shape, yellow-brown/grey-brown cell color, one kind of cortical granule, and two macronuclear nodules. The cortical granules in A. pulchra sp. n. are grouped in small clusters which are irregularly distributed both ventrally and dorsally; in A. candida sp. n., they are irregularly and sparsely distributed ventrally and form longitudinal stripes dorsally. The ontogenetic processes of both species show similarities to those of other congeners. Phylogenetic trees based on SSU rRNA gene sequences suggest that the family Amphisiellidae is a paraphyletic assemblage. The results further demonstrate that two isolates identified as Amphisiella annulata (DQ832260 and GU170843) are very likely cryptic species, and a sequence identified as A. milnei (DQ845293) may belong to a genus other than Amphisiella., (Copyright © 2012 Elsevier GmbH. All rights reserved.)

Members of the ciliate genus Frontonia are common colonizers of periphytic communities in aquatic biotopes. Recent studies indicate that their species diversity is higher than previously supposed. In this study the morphology and infraciliature of three new species, Frontonia sinica spec. nov., F. pusilla spec. nov., and F. elegans spec. nov., isolated from coastal waters of China, were investigated using live observation and silver impregnation methods. Frontonia sinica differs from its congeners by the following combination of characters: ellipsoidal body, about 116 somatic and five or six vestibular kineties, peniculi 1 and 2 four-rowed, peniculus 3 two-rowed, and a single contractile vacuole. Frontonia pusilla has about 72 somatic kineties, four-rowed peniculi 1 and 2, a two-rowed peniculus 3, and two contractile vacuoles. Frontonia elegans has 73 somatic kineties, four-rowed peniculi 1 and 2, a three-rowed peniculus 3, and two contractile vacuoles. In the present work, six new small-subunit rRNA gene sequences of six Frontonia species are used to construct the phylogenetic trees. Our phylogenetic analysis supports that the genus Frontonia may be paraphyletic. Meanwhile, no pattern of correlation could be found between the structures of peniculi and the phylogenetic relationships of Frontonia species in the present study., (Copyright © 2012 Elsevier GmbH. All rights reserved.)

The morphology, infraciliature and small subunit ribosomal RNA gene-based phylogeny of an urostylid ciliate, Monocoronella carnea n. g., n. sp., found in coastal areas off Daya Bay, Southern China, were investigated. The new genus Monocoronella n. g. is recognized by the following features: having conspicuous frontal cirri forming a long and single corona; buccal and frontoterminal cirri present; single marginal row on each side; adoral zone, midventral complex and transverse cirri in Pseudokeronopsis mode. The type species M. carnea n. sp. is diagnosed by the combination of marine habitat and brown-reddish color of the cortical granules. Phylogenetic analyses for the new taxon indicate that Monocoronella n. g. is most closely related to Bergeriella, and is located within the core Urostylida clade. A misidentification in previous literature was recognized and a new species, Monocoronella dragescoi n. sp. [Basionym: Holosticha (Keronopsis) monilata (Kahl 1928) sensu Dragesco (1970) et sensu Dragesco and Dragesco-Kernéis (1986), non sensu Kahl (1928)], was suggested., (© 2011 The Author(s). Journal of Eukaryotic Microbiology © 2011 International Society of Protistologists.)

The morphology, morphogenesis, and phylogeny of an undescribed oligotrich sensu lato (s. lat.) ciliate, Lynnella semiglobulosa n. g., n. sp., found in Daya Bay, southern China, were investigated. This species shares some features with both oligotrichs sensu stricto and choreotrichs, but most morphological and morphogenetic characters as well as the phylogenetic analysis suggest that it should be assigned into subclass Choreotrichia temporarily. Lynnella semiglobulosa is distinguished from members of all known genera and families of the subclass Choreotrichia by a unique combination of characteristics of the buccal and somatic ciliatures. Thus, a new family Lynnellidae n. fam. and a new genus Lynnella n. g. are proposed for it. The new family is distinguished by an open adoral zone of membranelles (AZM) in which, however, there are no ventral membranelles; the distal and proximal portions of the new adoral zone lie close to each other forming an open circle in stomatogenesis. The new genus Lynnella is characterized by possessing two longitudinally oriented somatic kineties, one dorsal and one ventral, several proximal membranelles progressively lengthened toward the proximal end of adoral zone, and two macronuclear nodules. In phylogenetic analyses based on small subunit rRNA gene sequences, L. semiglobulosa clustered basally to all choreotrichs, but with relatively weak support; nevertheless, the possibility of a relationship with the subclass Oligotrichia was not rejected by the approximately unbiased nor Shimodaira-Hasegawa test. Based on morphological, morphogenetic, and molecular evidence of L. semiglobulosa, it is confirmed that the open AZM should be a plesiomorphic character of oligotrichs s. lat. as suggested previously., (© 2010 The Author(s). Journal of Eukaryotic Microbiology © 2010 International Society of Protistologists.)

The morphology, infraciliature, and small subunit (SSU) rRNA gene sequences of two new pleurostomatid ciliates, Epiphyllum shenzhenense n. sp. and Loxophyllum spirellum n. sp., isolated from a mangrove wetland near Shenzhen, South China, were investigated. Epiphyllum shenzhenense n. sp. is morphologically characterized by leaf-shaped cell about 150 x 35 microm in vivo, usually with four contractile vacuoles, 20-29 right kineties and 10-26 left kineties, ca. four macronuclear nodules, and two types of extrusomes (i.e. short spindle-shaped and long bar-shaped). As a new species, L. spirellum n. sp. is distinguished from its congeners by its posterior dorsal margin twisted onto the left side, the distribution of extrusomes (evenly arranged along the oral slit, the posterior end, and clustered to 7-13 warts on dorsal margin), the subterminally positioned contractile vacuole, the number of kineties (8-10 on right side, 4-5 on left side), and its genetic distance from congeners. Phylogenetic trees based on the SSU rRNA gene sequence for both organisms were constructed, which indicate that Epiphyllum is a distinct genus and occupies a basal position in the Pleurostomatida clade; L. spirellum n. sp. falls well into the Loxophyllum clade, which has a close relationship with Litonotus and Spiroloxophyllum.

The small subunit ribosomal RNA genes of nine species belonging to six genera of litostome ciliates, namely Amphileptus aeschtae, Chaenea teres, Chaenea vorax, Lacrymaria marina, Litonotus paracygnus, Loxophyllum sp.-GD-070419, Loxophyllum jini, Loxophyllum rostratum, and Phialina salinarum, were sequenced for the first time. Phylogenetic trees were constructed using different methods to assess the inter- and intra-generic relationships of haptorians, of which Chaenea, Lacrymaria, Litonotus, and Phialina were analyzed for the first time based on molecular data. Monophyly of the order Pleurostomatida was strongly confirmed, and the two existing families of pleurostomatids, created on the basis of morphology, were confirmed by molecular evidence. Within the Pleurostomatida, Siroloxophyllum utriculariae occupied a well-supported position basal to the Loxophyllum clade, supporting the separation of these genera from one another. Both the subclass Haptoria and the order Haptorida were partially unresolved, possibly paraphyletic assemblages of taxa in all analyses, creating doubts about the traditional placement of some haptorid taxa. The existing sequence of L. rostratum in GenBank (DQ411864) was conspicuously different from that of the isolate from Qingdao, China sequenced in the present work, indicating that they are different species. The isolate from Qingdao was verified as L. rostratum by morphological analysis, and the published morphology of existing GenBank record of L. rostratum is different from it. Based on both morphological and molecular evidence, the latter may be congeneric with an undescribed species of Loxophyllum from Guangdong Province, China.

The cortical development during binary fission of the relatively poorly known stichotrich ciliate, Trachelostyla pediculiformis (Cohn, 1866) Borror, 1972, found in coastal waters near Qingdao, China, was investigated using the protargol impregnation method. The morphogenetic process reveals some pretty unusual characteristics, which do not follow the Oxytricha-pattern: (1) the parental oral apparatus is entirely renewed from an oral primordium formed de novo in the proter; (2) in the proter, the parental undulating membranes are not involved in the formation of the newly formed oral primordium; both undulating membrane-anlagen (UM-anlage) and frontoventral-transverse cirral anlagen (FVT-anlagen) develop from the oral primordium in the proter; (3) the dorsal kineties (DK) are generated in a unique way, that is, in both dividers, two separate groups of DK-anlagen develop in the right- and left-most DK, generate all the DK and evolve to replace the old structures; (4) three caudal cirri are formed at the posterior ends of three right-most dorsal kinety anlagen; (5) eight frontal, five ventral and five transverse cirri are derived from six streaks, namely, the UM-anlage and 5 FVT-anlagen; the cirri are segregated from these anlagen in the pattern 1:3:3:3:4:4 (from left to right) in the Oxytricha mode. Based on both SSrRNA gene sequencing and morphogenetic data, the systematic positions of the genus Trachelostyla Borror, 1972 as well as the family Trachelostylidae Small and Lynn, 1985 are briefly analyzed. The results indicate that this genus/family could be a highly isolated lineage and might be ancestral to other well-known oxytrichids.

The "well-known" marine hypotrichous ciliate, Amphisiella marioni Gourret & Roeser, 1888 has been repeatedly misidentified for over a century, which has led to great confusion in the species identification. Based on a population collected from mariculture water in Qingdao (Tsingtao), north China, the morphology and infraciliature were investigated using live observations and the protargol impregnation method. The Chinese population corresponds perfectly with the original description by Gourret and Roeser (1888). As the type species of Amphisiella, its morphological characteristics include: an elongated body about 90-150 microm long in vivo; with grouped cortical granules arranged in irregular rows throughout the whole body; a contractile vacuole located slightly behind mid-body; adoral zone of membranelles slightly bipartite in structure with the anterior part ventrally located; amphisiellid median cirral row (ACR) extending to the level of the transverse cirri (TC); about eight frontal cirri left of the ACR; closely spaced marginal rows, displaced inwards, each with about 32 cirri; six TC arranged in a V-shape; no caudal cirrus; five to six dorsal kineties; and typically two macronuclei. We conclude that the populations described by Mansfeld (1923) and Wicklow (1982), respectively, were misidentified. To clarify the taxonomic status of this species, the population from the northern Chinese coast of the Yellow Sea is designated as a neotype.

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The ciliate genus Chlamydodon is characterized by a unique cross-striated band (CSB) along the cell perimeter. To date, more than 15 nominal species have been assigned to this genus, all of which are exclusively from marine or brackish water. In the present work, we have revised the genus according to the available data and suggest an illustrated key to aid species diagnosis. In addition, the systematic relationships of chlamydodontid congeners were analysed based on SSU rRNA gene sequences, indicating that all congeners belong to a well-defined clade. Furthermore, we investigated three species from coastal areas of China, including two new species, Chlamydodon wilberti sp. nov. and C. bourlandi sp. nov., using morphological and phylogenetic criteria. Chlamydodon wilberti sp. nov. is characterized by a cell size of 65-105  ×  35-60 µm, a complete CSB, and 38-49 somatic kineties. Chlamydodon bourlandi sp. nov. has a cell size of 150-250  ×  65-150 µm, a complete CSB, a reddish to violet cell colour, 83-97 somatic kineties, and 40-68 contractile vacuoles. As a further contribution, a well-studied species, C. triquetrus (Müller, 1786) Kahl, 1931, is also re-described. www.zoobank.org/urn:lsid:zoobank.org:pub:B83A5466-9D2B-4502-9A23-F16A61D48172. [ABSTRACT FROM AUTHOR]

Abstract: Three new cyrtophorian ciliates isolated from coastal areas of China were described based on morphological and genetic data. The Chlamydodon mnemosyne‐like species Chlamydodon similis sp. n. differs from its congeners mainly by its number of somatic kineties. Chlamydodon oligochaetus sp. n. is distinguished from its congeners mainly by having fewer somatic kineties, and/or an elongated body shape. Chlamydodon crassidens sp. n. is characterized mainly by an inverted triangular body shape, a posteriorly interrupted cross‐striated band (5–6 μm wide), and a large cytostome. Moreover, we provided small‐subunit (SSU) rDNA sequences of C. similis sp. n. and C. oligochaetus sp. n. Maximum likelihood (ML) and Bayesian inference (BI) consistently placed C. similis sp. n. as a sister to C. paramnemosyne, but showed different branching position of C. oligochaetus sp. n., which may be due to a low taxon sampling in the Chlamydodontidae and/or an insufficient resolution of the marker gene at species level. [ABSTRACT FROM AUTHOR]

A high diversity of pleurostomatid ciliates has been discovered in the last decade, and their systematics needs to be improved in the light of new findings concerning their morphology and molecular phylogeny. In this work, a new genus, Protolitonotus gen. n., and two new species, Protolitonotus magnus sp. n. and Protolitonotus longus sp. n., were studied. Furthermore, 19 novel nucleotide sequences of SSU rDNA, LSU rDNA and ITS1-5.8S- ITS2 were collected to determine the phylogenetic relationships and systematic positions of the pleurostomatid ciliates in this study. Based on both molecular and morphological data, the results demonstrated that: (i) as disclosed by the sequence analysis of SSU rDNA, LSU rDNA and ITS1-5.8S- ITS2, Protolitonotus gen. n. is sister to all other pleurostomatids and thus represents an independent lineage and a separate family, Protolitonotidae fam. n., which is defined by the presence of a semi-suture formed by the right somatic kineties near the dorsal margin of the body; (ii) the families Litonotidae and Kentrophyllidae are both monophyletic based on both SSU rDNA and LSU rDNA sequences, whereas Amphileptidae are non-monophyletic in trees inferred from SSU rDNA sequences; and (iii) the genera Loxophyllum and Kentrophyllum are both monophyletic, whereas Litonotus is non-monophyletic based on SSU rDNA analyses. ITS1-5.8S- ITS2 sequence data were used for the phylogenetic analyses of pleurostomatids for the first time; however, species relationships were less well resolved than in the SSU rDNA and LSU rDNA trees. In addition, a major revision to the classification of the order Pleurostomatida is suggested and a key to its families and genera is provided. [ABSTRACT FROM AUTHOR]

Two new pleurostomatid ciliates, Loxophyllum lembum sp. n., L. vesiculosum sp. n., and the poorly known L. perihoplophorum Buddenbrock, 1920, isolated from brackish waters in coastal regions of southern China, are described following observations of live cells and protargol-impregnated specimens. Loxophyllum lembum sp. n. is distinguished by a combination of characters including two macronuclear nodules, 6-9 contractile vacuoles along the ventral margin, 11-14 right and 6-8 left kineties and the presence of cortical granules. Loxophyllum vesiculosum sp. n. differs from its congeners mainly by the unique distribution of contractile vacuoles, several of which lie along the dorsal margin and one on the ventral margin, and 15-21 right and 6-8 left kineties. Loxophyllum perihoplophorum is characterized by its large cell size (350-450 μm long in vivo), 3-5 contractile vacuoles along the dorsal margin in the posterior region of the body, and 19-23 right and 7-9 left kineties. An improved diagnosis of L. perihoplophorum is provided. The SSU rDNA sequence of L. perihoplophorum is reported for the first time and its molecular phylogeny is analyzed. Maximum likelihood and Bayesian inference analyses of SSU rDNA sequence data recover the monophyly both of the order Pleurostomatida and of the genus Loxophyllum. [ABSTRACT FROM AUTHOR]

Abstract: We investigate the living morphology, infraciliature, and small subunit rRNA gene sequences of two poorly known marine euplotids, Aspidisca fuscaKahl, 1928 and A. hexerisQuennerstedt, 1869, isolated from intertidal sewage outfall, northern China and sandy sediments, southern China, respectively. Improved diagnoses and morphometric data are provided for both species. Aspidisca fusca is characterized by having a peristomial spur, seven frontoventral cirri in a “polystyla-arrangement”, and two oval macronuclear nodules. Aspidisca hexeris can be recognized by having an elliptical body shape, a peristomial spur, seven membranelles in the anterior portion of adoral zone and 11–13 in the posterior part, as well as seven frontoventral cirri arranged in two oblique rows. In addition, the presence and degree of projections along the left margin of A. hexeris are highly variable among populations. Phylogenetic analyses based on small subunit rRNA gene sequence data support the validity of both A. hexeris and A. fusca as distinct species, and indicate that A. fusca is most closely related to A. aculeata and A. steini, whereas A. hexeris shows a close relationship with A. magna. [Copyright &y& Elsevier]

Three new pleurostomatid ciliates, Loxophyllum meridionale sp. n., L. salinum sp. n., and L. planum sp. n., isolated from brackish waters of mangrove wetlands in southern China, were investigated using live observation and protargol impregnation. The main characteristic feature of L. meridionale sp. n. is the presence of three contractile vacuoles (CVs) along its dorsal margin. Loxophyllum salinum sp. n. can be identified by the presence of three closely spaced CVs along the ventral margin and 12-15 right and four to six left kineties. Loxophyllum planum sp. n. is characterized by the combination of two macronuclei, a single, subterminal CV, and the absence of cortical granules. Phylogenetic trees based on the small subunit r DNA sequence data recovered Loxophyllum as a monophyletic group within the pleurostomatids. Loxophyllum meridionale sp. n., L. salinum sp. n., and L. planum sp. n. were all nested within the Loxophyllum clade. Based on its ciliary pattern and general morphological features, Loxophyllum asetosum is transferred to the genus Amphileptus and a new combination is suggested: Amphileptus asetosus (Burkovsky 1970) comb. n. [basionym L. asetosum Burkovsky 1970]. [ABSTRACT FROM AUTHOR]

Recent studies indicate that there is a high diversity of pleurostomatid ciliates in the coastal waters of China. Here, three new congeners of Loxophyllum, L. caudatum sp. n., L. rugosum sp. n., and L. chinense sp. n., are described following observations of live cells and protargol-impregnated specimens. All three species usually have two macronuclear nodules and prominent warts along the dorsal margin formed by clustered extrusomes. In addition, L. caudatum sp. n. is characterized by its long conspicuous tail, dot-like cortical granules, 4 or 5 left and 9 or 10 right kineties, and a single subterminal contractile vacuole. Loxophyllum rugosum sp. n. is distinguished by possessing three prominent ridges on the left side, 7-11 right and 5-7 left kineties. Loxophyllum chinense sp. n. is characterized by having several contractile vacuoles distributed along the ventral margin, 13-18 right and 6-8 left kineties. The small subunit ribosomal DNA ( SSU rDNA) sequence similarities among six congeners range from 96.46% to 99.94%. Phylogenetic trees based on the SSU rDNA sequences indicate that all Loxophyllum spp. form a well-supported monophyletic group. A brief review of the marine and brackish Loxophyllum species is supplied and one new combination, Litonotus multiplicatus (Kahl 1931) comb. n. (basionym Loxophyllum multiplicatum Kahl 1931), and one new name, Litonotus dragescoi nom. n. (basionym L. fasciolatus Dragesco 1966), are suggested. [ABSTRACT FROM AUTHOR]

Cyrtophorids are a specialized group of ciliated protozoa with multitudinous morphotypes. In the present work, the morphology and infraciliature of two new and three rarely known species, including two new genera of cyrtophorid ciliates, Heterohartmannula fangi gen. et sp. nov., Aporthotrochilia pulex (Deroux, 1976) gen. et comb. nov., Trochilia alveolata sp. nov., Trochochilodon flavus Deroux, 1976, and Hypocoma acinetarum Collin, 1907, are described. Heterohartmannula gen. nov. is mainly characterized by a combination of features: two circumoral kineties obliquely arranged, podite not surrounded by somatic kineties, and no distinct gap between left and right ciliary field. Aporthotrochilia gen. nov. is diagnosed mainly by: podite present, oral ciliature reduced to two fragments, several kinety fragments positioned on the right posterior of frontoventral kineties and several terminal fragments. Phylogenetic analyses based on the small subunit rRNA (SSU rRNA) gene sequences support the establishment of two new genera and indicate that Heterohartmannula is most closely related to Hartmannula, and Aporthotrochilia is basal to the Cyrtophoria-Chonotrichia clade. Trochilia alveolata sp. nov. differs from its congeners mainly by having a conspicuous alveolar layer. In addition, detailed live and infraciliature data of Hypocoma acinetarum and Trochochilodon flavus are supplied. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 164, 1-17. [ABSTRACT FROM AUTHOR]

The morphology and small subunit rRNA (SSrRNA) gene sequence of a marine oligotrich ciliate, Williophrya maedai gen. nov., sp. nov., are reported. The new genus Williophrya is characterized by the adoral zone with no differentiation of membranelles, and the reduced somatic ciliature which comprises a bipartite girdle kinety only. In addition, the in vivo morphologies of two other oligotrichs, namely Strombidium basimorphum Martin & Montagnes, 1993 and Pseudotontonia simplicidens (Lynn & Gilron, 1993) Agatha, 2004, are reported for the first time based on Chinese populations. Improved diagnoses of both species are supplied. The phylogenetic position of Williophrya maedai is investigated based on SSrRNA gene sequence data. These show that: (1) Williophrya maedai is most closely related to Strombidium purpureum and S. apolatum; (2) Williophrya is assigned to the family Strombidiidae although it has some unique morphological features regarding its oral and somatic ciliatures. [ABSTRACT FROM AUTHOR]

The marine ciliate Nothoholosticha is characterized by having a Holosticha-like ciliature pattern but without migratory frontoterminal cirri. However, its systematic position and its relationship to other members of the Holosticha-complex have not yet been resolved. In order to gain deeper insights into these relationships, the small subunit rRNA (SSrRNA) gene and the rRNA internal transcribed spacer and 5.8SrRNA (ITS1-5.8S-ITS2) region of two marine Anteholosticha species, as well as the ITS1-5.8S-ITS2 region of N. fasciola, were sequenced, and molecular trees (BI, ML, NJ and MP trees) were constructed. Although our analyses failed to conclusively resolve the phylogeny of this assemblage, certain conclusions could be drawn. Firstly, Nothoholosticha is a valid genus that is more closely related to Pseudokeronopsis than to other Holosticha-complex genera. Secondly, sequence analyses and phylogenetic trees of several Anteholosticha species revealed a high molecular diversity, which does not support the monophyly of this genus. Thirdly, the current assignment of certain well-known genera, e.g. Holosticha, Anteholosticha, Apokeronopsis, Parabirojimia, Psammomitra, Diaxonella, Metaurostylopsis and Thigmokeronopsis, to the families Bakuellidae (sensu Berger 2006), Urostylidae (sensu Berger 2006) or Holostichidae (sensu Berger 2006) is challenged by the molecular data presented here. And fourthly, the families Holostichidae and Pseudokeronopsidae (sensu Lynn 2008) are probably paraphyletic, and their systematic assignments await further evaluation. [ABSTRACT FROM AUTHOR]

Ciliates possess two distinct nuclear genomes and unique genomic features, including highly fragmented chromosomes and extensive chromosomal rearrangements. Recent transcriptomic surveys have revealed that ciliates have several multi-copy genes providing an ideal template to study gene family evolution. Nonetheless, this process remains little studied in ciliated protozoa and consequently, the evolutionary patterns that govern it are not well understood. In this study, we focused on obtaining fine-scale information relative to ciliate species divergence for the first time. A total of 230 actin gene sequences were derived from this study, among which 217 were from four closely related Pseudokeronopsis species and 13 from other hypotrichous ciliates. Our investigation shows that: (1) At least three duplication events occurred in ciliates: diversification of three actin genes (Actin I, II, III) happened after the divergence of ciliate classes but before that of subclasses. And several recent and genus-specific duplications were followed within Actin I ( Sterkiella , Oxytricha , Uroleptus , etc.), Actin II ( Sterkiella ), respectively. (2) Within the genus Pseudokeronopsis , Actin I gene duplication events happened after P. carnea and P. erythrina diverged. In contrast, in the morphologically similar species P. flava and P. rubra , the duplication event preceded diversification of the two species. The Actin II gene duplication events preceded divergence of the genus Pseudokeronopsis . (3) Phylogenetic analyses revealed that actin is suitable for resolving ciliate classes, but may not be used to infer lower taxon relationships. [ABSTRACT FROM AUTHOR]