Evolution of ciliary patterns in the Oligotrichida (Ciliophora, Spirotricha) and its taxonomic implications

Although the somatic ciliature of the Oligotrichida typically comprises only a girdle and ventral kinety, a considerable diversity of ciliary patterns occurs. The four main girdle kinety patterns are identically found in tailed and tail-less species. The contractile tail has a complicated and unique ultrastructure and is potentially useful for the cell's movement and/or stabilization during feeding. Accordingly, I assume that this structure has evolved only once, namely, in the Tontoniidae nov. fam., and that the different girdle kinety patterns developed convergently in the tailed and tail-less taxa. Further distinct features suggest the establishment of the families Cyrtostrombidiidae nov. fam. (with cyrtos-like pharyngeal fibres and lack of ventral membranelles and endoral) and Pelagostrombidiidae nov. fam. (with neoformation organelle). An attempt is made to reconstruct the evolution of the kinety patterns based on morphologic, ontogenetic, and ultrastructural data. Some genera of tail-less Oligotrichida base on differences in the ciliary pattern; Omegastrombidium nov. gen. is erected for a further girdle kinety pattern. Likewise, the tailed genus Tontonia is split, resulting in two new genera, viz., Pseudotontonia nov. gen. and Spirotontonia nov. gen. Furthermore, the genus Spirostrombidium is split due to the different origin of the parallel course of girdle and ventral kinety, and Parallelostrombidium nov. gen. is established. However, the genus Thigmostrombidium is rejected because its enlarged thigmotactic membranelles are interpreted as an adaptation to the benthic lifestyle, which occurred several times within different girdle kinety patterns.     

Pelagostrobilidium liui n. sp. (Ciliophora,Choreotrichida) from the Coastal Waters of Northeastern Taiwan and an Improved Description of Pelagostrobilidium minutum Liu et al., 2012

The number of somatic kineties in Pelagostrobilidium ranges from 4 to 6 according to the present state of knowledge. This study investigates Pelagostrobilidium liui n. sp. using live observation, protargol stain, and small subunit rDNA data sequencing. Pelagostrobilidium liui n. sp. is characterized by having a spherical‐shaped body, four somatic kineties, with kinety 2 spiraled around the left side of body, about six elongated external membranelles, and invariably no buccal membranelle. It differs from its most similar congener, Pelagostrobilidium minutum Liu et al., 2012 , in (i) cell shape; (ii) macronucleus width; (iii) oral apparatus; (iv) anterior orientation of kinety 2; (v) location where kinety 2 commences; (vi) arrangement of kinety 1; (vii) distance between the anterior cell end and the locations where kineties commence; and (viii) the presence of 12 different bases (including two deletions) in the small subunit rDNA sequences. The diagnosis of P. minutum Liu et al., 2012 is also improved to include the following new characteristics: invariably four somatic kineties; kineties 2 and 4 alone commence at the same level; kinety 2 originates from right anterior cell half on ventral side, extends sinistrally posteriorly, over kinety 1, around left posterior region, terminates near posterior cell end on dorsal side; kinety 1 commences below anterior third of kinety 2.     

Updated hypothesis on the evolution of oligotrichid ciliates (Ciliophora, Spirotricha, Oligotrichida) based on somatic ciliary patterns and ontogenetic data

The two recently established genera ApostrombidiumXu et al., 2009 and VaristrombidiumXu et al., 2009 and the analysis of ontogenetic data in Strombidium constrictum, S. montagnesi, S. wilberti, Omegastrombidium elegans, and Paratontonia gracillima necessitated a revision of the hypothesis about the somatic ciliary pattern evolution in oligotrichid ciliates. As a consequence, the species-rich genus Strombidium was split, establishing two genera for species with a horizontal girdle kinety posterior to the oral primordium: Opisthostrombidium nov. gen. with the extrusome attachment sites along the anterior margin of the girdle kinety and posterior to the oral primordium and Foissneridium nov. gen. with the extrusome attachment sites distinctly apart from the girdle kinety and anterior to the oral primordium. The ontogenetic data revealed that the Ω-shaped girdle kinety pattern evolved convergently from the Pseudotontonia pattern with its horizontal girdle kinety in the tailed genus Paratontonia and from the Novistrombidium pattern with its dextrally spiralled girdle kinety in the tailless genus Omegastrombidium. The somatic ciliary pattern of the latter genus probably gave rise to the patterns of Apostrombidium and Varistrombidium.     

New contributions to the Cyrtophoria ciliates (Protista,Ciliophora): Establishment of new taxa and phylogenetic analyses using two ribosomal genes

Periphytic ciliates play a vital role in the material cycle and energy flow of microbial food web, however, their taxonomy and biodiversity are inadequately studied given their high species richness. Two new and one little known species, viz. Derouxella lembodes gen. et sp. nov., Cyrtophoron multivacuolatum sp. nov., and Cyrtophoron apsheronica Aliev, 1991, collected from coastal waters of China, were investigated using modern methods. Derouxella gen. nov. can be recognized by having dorsoventrally flattened body, a podite, one fragmented preoral kinety, two parallel circumoral kineties, and somatic kineties progressively shortened from right to left. Morphological classification and phylogenetic analyses based on nuclear small subunit ribosomal RNA (nSSU rRNA) and mitochondrial small subunit ribosomal RNA (mtSSU rRNA) gene sequence data inferred that Derouxella gen. nov. occupies an intermediate position between Hartmannulidae and Dysteriidae. Cyrtophoron multivacuolatum sp. nov. is characterized by large body size, the numbers of somatic kineties and nematodesmal rods, and having numerous contractile vacuoles. The genus Cyrtophoron and the poorly known species C. apsheronica were redefined. Even with the addition of newly obtained nSSU rRNA and mtSSU rRNA gene sequences of Cyrtophoron, the family Chlamydodontidae was still recovered as a monophyletic group, the monophyly of Cyrtophoron was supported too.     

A new oligotrich (Ciliophora,Oligotrichia) from Argentina,with redefinition of Novistrombidium Song and Bradbury

A new oligotrich similar to Novistrombidium was discovered in plankton samples from an artificial tributary of the Salado River, Buenos Aires province, Argentina, in summer 2010. Propecingulum fistoleramalliei sp. n. has an obovate and anteriorly truncated body, with a conspicuous ventral furrow, is flattened ventrally, and has a prominent right apical protrusion. It temporarily attaches to the substratum by a posterior mucous thread. Rod-shaped extrusomes arranged equidistantly and insert directly above the girdle kinety. The macronucleus is globular to ellipsoidal. The contractile vacuole is located in the left, anterior quarter of the cell and the adoral zone is composed of 30–35 collar, 9–14 buccal, and two thigmotactic membranelles. The girdle kinety is dextrally spiraled and ventrally open; the ventral kinety is posterior to anterior end of the girdle kinety. The oral primordium develops posterior to the right thigmotactic membranelle and anterior the stripe of extrusomes above left, lateral portion of the girdle kinety. The SSUrDNA phylogeny confirms one more time that Novistrombidium is not monophyletic; consequently, we elevate the subgenus Propecingulum up to genus rank and redefine the genus Novistrombidium.     

Reconciling cladistic and genetic analyses in choreotrichid ciliates (Ciliophora, Spirotricha, Oligotrichea)

Fifty-six features of halteriid, oligotrichid, and choreotrichid ciliates are cladistically analysed, including an updated hypothesis about the evolution of the somatic ciliary patterns. Based on its morphology, Lynnella clusters with Parastrombidinopsis, Parastrombidium, and Strombidinopsis, while it is basal to the other choreotrichids in the molecular phylogenies. The two clusters of Favella species in small subunit rRNA gene trees are supported by morphological features, justifying a separation at genus and family level. The genus Favella has a smooth lorica surface and a somatic ciliary pattern comprising a left and lateral ciliary field as well as two dorsal kineties and a monokinetidal ventral kinety abutting on the right ciliary field. The new genus Schmidingerella n. gen., established for the second Favella cluster, groups with Metacylis and Rhabdonella in the molecular trees. It differs from Favella in (i) a lorica wall with reticulate surface ridges and minute openings and (ii) a ventral kinety that is distinctly apart from the right ciliary field and composed of a monokinetidal anterior and a dikinetidal posterior portion. The genera Codonaria, Codonella, and Codonellopsis are affiliated with the family Dictyocystidae, whose diagnosis is improved to include the lorica sac.     

Three new entocommensal ciliates from digestive tract of sea urchins of the Weddell Sea,Antarctica (Protozoa,Ciliophora)

Three new entocommensal ciliates (Spirostrombidium echini sp. nov., Entodiscus pseudoechini sp. nov. and Cryptochilum antarcticum sp. nov.) from the digestive tract of sea urchins in Antarctica (Weddell Sea) were studied using protargol and silver nitrate impregnation methods. Based on our reinvestigations, improved diagnoses for the genera Entodiscus and Cryptochilum are provided. According to the general morphology, E. pseudoechini is recognized by a higher number of ciliary rows, the absence of a caudal cilium which is typically present for other congeners, and shortened somatic kineties direct to the ventral surface, of which the last somatic kinety does not pass through the caudal pole as in most other scuticociliates. C. antarcticum differs from the known congeners in its particularly high number of somatic kineties. S. echini is distinguished by the combination of habitat, body shape, structure of buccal apparatus and arrangement of somatic kineties. Accepted: 29 March 1999     

Agolohymena aspidocauda nov. gen., nov. spec., a histophagous freshwater tetrahymenid ciliate in the family Deltopylidae (Ciliophora,Hymenostomatia), from Idaho (northwest U.S.A.): Morphology,ontogenesis and molecular phylogeny

Morphology, ontogeny and the molecular phylogeny of Agolohymena aspidocauda nov. gen., nov. spec., a new freshwater tetrahymenid ciliate from Idaho, U.S.A, are described. The ontogeny and histophagous mode of nutrition are similar to those of Deltopylum rhabdoides Fauré-Fremiet and Mugard, 1946. The new genus is placed with Deltopylum in the resurrected family Deltopylidae Song and Wilbert, 1989. We emend the diagnostic features of the family to include division by polytomy, right and left somatic kineties extending into the preoral suture, crook-shaped or sigmoid adoral membranelles 1 and 2, markedly reduced adoral membranelle 3 and a tetrahymenid silverline pattern. The main diagnostic features of the new genus are a disc-shaped caudal ciliary array and formation of two types of resting cysts, one smooth and the other bearing tangled tubular or cylindrical lepidosomes. Nuclear small subunit ribosomal RNA gene and mitochondrial cytochrome oxidase subunit 1 gene sequences place the new genus basal within the order Tetrahymenida, well separated from members of the family Tetrahymenidae (Lambornella and Tetrahymena) and also from other tetrahymenids (Colpidium, Dexiostoma, Glaucoma). The genetic divergences between this species and other genera in Tetrahymenida are large enough to suggest placement of the new genus in a separate family. This corroborates the morphological data, since the elaborate caudal ciliary array and the lepidosome-covered resting cyst of this species are not found in other Tetrahymenidae.     

Morphology,Morphogenesis and Molecular Phylogeny of a New Obligate Halophile Ciliate,Schmidtiella ultrahalophila gen. nov., spec. nov. (Ciliophora,Hypotrichia) Isolated from a Volcanic Crater on Sal (Cape Verde Islands)

A new hypotrichous ciliate, Schmidtiella ultrahalophila gen. nov., spec. nov., was isolated from a solar saltern on the island of Sal, Cape Verde. The possession of only one short dorsal kinety clearly distinguishes S. ultrahalophila from other known hypotrichous genera and species. Further diagnostic characters include: a flexible and slender body, an average size of 85 × 15 μm in vivo; a bipartite adoral zone with two hypertrophied frontal adoral membranelles and nine to twelve ventral adoral membranelles; three frontal, one parabuccal, two frontoventral, two or three postoral ventral, and two or three frontoterminal cirri; and marginal cirral rows variable in number, usually one on each side. Ontogenetic data indicate the following: the frontal‐ventral cirri originate from six or five anlagen; the proter inherits the parental adoral zone; the frontal and ventral cirri originate from five or six anlagen; and the marginal cirral rows and the dorsal kinety tend to originate intrakinetally. Additional marginal rows are rarely derived from de novo anlagen. Based on its morphology, morphogenesis and its SSU rRNA phylogenetic placement, the new species should be assigned to the order Sporadotrichida Fauré‐Fremiet, 1961. Due to low taxon sampling, however, its exact position in this order remains enigmatic.     

Taxonomy of five species of cyrtophorids (Protozoa: Ciliophora) including consideration of the phylogeny of two new genera

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.     

A barotolerant ciliate isolated from the abyssal deep sea of the North Atlantic: Euplotes dominicanus sp. n. (Ciliophora,Euplotia)

A new Euplotes species, isolated from abyssal depths (>4000 m) of the North Atlantic Ocean, was described based on morphology, ciliary pattern and molecular data. Euplotes dominicanus sp. n. is characterized by a small body size (29–40 × 17–27 μm in vivo), 18–22 adoral membranelles, 10 frontoventral, five transverse and two left marginal cirri and one caudal cirrus, five or six dorsolateral kineties with 7–9 dikinetids in mid-dorsolateral kinety (DK3), and dorsal silverline system of the double-eurystomus type. Phylogenetic analyses inferred from 18S rRNA sequences show that Euplotes dominicanus sp. n. is most closely related to E. curdsi, with a sequence similarity of 97.6 %. Euplotes dominicanus sp. n. was able to survive hydrostatic pressures up to 500 bar indicating its barotolerance. Metabarcoding data demonstrate the presence of E. dominicanus sp. n. in sediments of several deep-sea basins.     

Novistrombidium sinicum n. sp. and Novistrombidium orientale n. sp. (Protozoa: Ciliophora): Two New Oligotrich Ciliates from a Mangrove Wetland, South China

ABSTRACT. The morphology and infraciliature of two new marine oligotrich ciliates, Novistrombidium sinicum n. sp. and Novistrombidium orientale n. sp., isolated from a mangrove wetland near Guangzhou, southern China, were studied from live and protargol-stained specimens. Novistrombidium sinicum is different from its congeners by the combination of the following characters: three posteriorly directed thigmotactic membranelles, one ellipsoidal macronucleus, the extrusomes equidistantly arranged, and the ventral kinety commencing below the right end of the girdle kinety. The small-sized N. orientale can be separated from its congeners by two posteriorly directed thigmotactic membranelles, one ellipsoidal macronucleus, the extrusomes equidistantly arranged, and the ventral kinety commencing below the right end of the girdle kinety.     

Morphology and molecular phylogeny of two new species of Spirostrombidium (Ciliophora,Oligotrichia), with a key to the species of Spirostrombidium

Microscopic methods were used to investigate the morphological characterization of two novel oligotrich ciliates, Spirostrombidium paraurceolare sp. nov. and Spirostrombidium faurefremieti sp. nov., isolated from a mangrove wetland in Zhanjiang and an intertidal sandy beach in Qingdao, respectively. Spirostrombidium paraurceolare sp. nov. is characterized by three thigmotactic and 8–10 buccal membranelles, the girdle kinety spiralling around cell with one and a half whorls, and located at right anterior third of dorsal side anteriorly. Spirostrombidium faurefremieti sp. nov. can be recognized by a prominently deep and broad buccal cavity, two thigmotactic and 15–19 buccal membranelles, and the girdle kinety spiralling around cell with two whorls. The small subunit ribosomal RNA genes of these two species were sequenced and compared with those of their congeners to reveal nucleotide differences. Phylogenetic analyses revealed that the genus Spirostrombidium is non-monophyletic. Spirostrombidium faurefremieti sp. nov. falls into a clade comprising most congeners, but Spirostrombidium paraurceolare sp. nov. branches off and groups with Varistrombidium kielum with moderate support. A key to the identification of Spirostrombidium species is also provided.www.zoobank.org/urn:lsid:zoobank.org:pub:AB96BEE6-BE3A-4B95-B75A-3469B1C53ABB2     

The apertospathulidae, a new family of haptorid ciliates (protozoa, ciliophora)

We studied the morphology of three rare haptorid ciliates, using live observation and silver impregnation: Apertospathula verruculifera n. sp., Longispatha elegans n. gen., n. sp., and Rhinothrix porculus (Penard, 1922) n. gen., n. comb. Simple ethanol fixation (50-70%, v/v) is recommended to reveal the ciliary pattern of "difficult" ciliates, such as R. porculus, by protargol impregnation. The three genera investigated have a distinct feature in common, viz., a lasso-shaped oral bulge and circumoral kinety, where the right half is slightly to distinctly longer than the left and the circumoral kinety is open ventrally. Thus, they are united in a new spathidiid family, the Apertospathulidae n. fam., which probably evolved from a Bryophyllum-like ancestor by partial reduction of the oral bulge and circumoral kinety. Apertospathula verruculifera has a wart-like process, the palpus dorsalis, at the anterior end of the dorsal brush. The right branch of the circumoral kinety is only slightly longer than the left one. Longispatha elegans has a straight oral bulge and circumoral kinety, the right branch of which extends to the posterior end of the body while the left branch ends in the anterior third of the body. Rhinothrix porculus, a curious ciliate with a snout-like dorsal elongation of the oral bulge, the palpus oralis, has a highly characteristic ciliary pattern: the oral pattern is as in Longispatha, but the bulge and circumoral kinety extend spirally to the posterior end of the body while the somatic kineties course meridionally. This is achieved by inserting some shortened kineties in the curves of the oral bulge.     

Taxonomy and molecular systematics of three oligotrich (s.l.) ciliates including descriptions of two new species,Strombidium guangdongense sp. nov. and Strombidinopsis sinicum sp. nov. (Protozoa,Ciliophora)

In this study we investigated the morphology of three oligotrich (s.l.) ciliates, Strombidium guangdongense sp. nov., Cyrtostrombidium paralongisomum Tsai et al., 2015 and Strombidinopsis sinicum sp. nov. Strombidium guangdongense sp. nov. is characterized by its elongate obconical to obovoidal body shape and widely spaced dikinetids in the girdle and ventral kineties. Another new species, Strombidinopsis sinicum sp. nov. is diagnosed by its small size and semi-globular body shape without mineral envelopes. Some additional morphological data of the recently described species Cyrtostrombidium paralongisomum Tsai et al., 2015, such as the endoral membrane, are supplied based on our population. We also analysed the molecular phylogeny of each species based on small subunit rRNA (SSU rRNA) gene sequence data. The monophyly of Cyrtostrombidium is supported by our phylogenetic analyses, but the monophyly of Strombidinopsis and of the family Strombidinopsidae are both rejected by AU tests. In addition, Strombidium species have a tail branch separately from one another in phylogenetic trees, whereas strombidiids with a pigment spot group together, suggesting the latter character is a synapomorphy for this group of strombidiids.

http://zoobank.org/urn:lsid:zoobank.org:act:35FE2AFD-A582-4885-BD01-48901E4C76C4

http://zoobank.org/urn:lsid:zoobank.org:act:E9D4A497-DAD6-4AA0-A044-A5A1D2C1A057     

Three new anascosporic genera of the Saccharomycotina: Danielozyma gen. nov., Deakozyma gen. nov. and Middelhovenomyces gen. nov

Three new non-ascosporic, ascomycetous yeast genera are proposed based on their isolation from currently described species and genera. Phylogenetic placement of the genera was determined from analysis of nuclear gene sequences for D1/D2 large subunit rRNA, small subunit rRNA, translation elongation factor-1α and RNA polymerase II, subunits B1 and B2. The new taxa are: Deakozyma gen. nov., type species Deakozyma indianensis sp. nov. (type strain NRRL YB-1937, CBS 12903); Danielozyma gen. nov., type species Danielozyma ontarioensis comb. nov. (type strain NRRL YB-1246, CBS 8502); D. litseae comb. nov. (type strain NRRL YB-3246, CBS 8799); Middelhovenomyces gen. nov., type species Middelhovenomyces tepae comb. nov. (type strain NRRL Y-17670, CBS 5115) and M. petrohuensis comb. nov. (type strain NRRL Y-17663, CBS 8173).     

Two new marine scuticociliates,Sathrophilus planus n. sp. and Pseudoplatynematum dengi n. sp., with improved definition of Pseudoplatynematum (Ciliophora,Oligohymenophora)

Two new marine scuticociliates, Sathrophilus planus n. sp. and Pseudoplatynematum dengi n. sp., isolated from coastal waters of Qingdao, northern China, were investigated using live observation and silver impregnations. Sathrophilus planus can be recognized by its elongate and conspicuously flattened body, ca. 16 somatic kineties, single postoral kinety, extremely elongated first kinety row of membranelle 1, and its marine habitat. Among these features, the structure of membranelle 1 is the most characteristic and enables this species to be easily distinguished from its congeners. Pseudoplatynematum dengi is characterized by the following features: cell surface conspicuously strengthened and notched; prominent spines both at anterior and posterior ends of cell; membranelles 1 and 2 three-rowed, membranelle 3 single-rowed; paroral membrane composed of two parts, anterior end of upper part extending to about level of membranelle 2; ca. 20 somatic kineties, one postoral kinety; single prolonged caudal cilium; contractile vacuole caudally positioned. The diagnosis of the genus Pseudoplatynematum was renewed to include some new characteristics revealed by silver impregnation methods.     

Morphology and infraciliature of two new marine ciliates, Paracyrtophoron tropicum nov. gen., nov. spec. and Aegyria rostellum nov. spec. (Ciliophora, Cyrtophorida), isolated from tropical waters in southern China

The morphology and infraciliature of two new marine cyrtophorid ciliates, Paracyrtophoron tropicum nov. gen., nov. spec. and Aegyria rostellum nov. spec., isolated from tropical waters in southern China, were investigated using live observation and protargol impregnation methods. Paracyrtophoron nov. gen. differs from the closely related Cyrtophoron by lack of fragment kinety at anterior ends of right somatic kineties and thigmotactic cilia in posterior portion of ventral surface, while from the well-defined Chlamydodon by lack of the cross-striped band around the periphery of the somatic field. Paracyrtophoron tropicum nov. spec., the type of the new genus, can be recognized by the combination of the following characters: cell size about 150-175×70-90μm in vivo; elliptical to kidney-shaped in outline, dorsoventrally flattened about 2.5:1; conspicuous cortical granules; one canal-like depression extending from postoral area to subcaudal region of cell; ca. 90 somatic kineties; 12-16 nematodesmal rods; one or two terminal fragments on dorsal side. Aegyria rostellum is characterized by the following features: size about 90-150×40-70μm in vivo, triangular or ear-shaped body with broad anterior end, having a rostriform structure and pigment spots, 56-63 somatic kineties, one preoral kinety, three or four circumoral kineties, and 32-42 nematodesmal rods. Based on previous and current studies, the definition for the genus Aegyria is updated: body dorsoventrally flattened; oral ciliature consisting of one preoral and several circumoral kineties; podite located in posterior ventral region and surrounded by somatic kineties; no obvious gap between right and left somatic kineties; postoral and left somatic kineties progressively shortened posteriorly from right to left. Additionally, two new combinations were proposed.     

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. glaucoma Müller, 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.