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.     

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.     

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.     

Morphology and molecular phylogeny of three new oligotrich ciliates (Protozoa,Ciliophora) from the South China Sea

The present paper documents the morphology and systematic positions of three new oligotrich ciliates, P arallelostrombidium obesum sp. nov. , P arallelostrombidium ellipticum sp. nov. , and S trombidium tropicum sp. nov. , which were sampled from habitats with different salinities in southern China. P arallelostrombidium obesum sp. nov. is characterized by a fat body and the posterior portions of the girdle and ventral kineties extending transversely on the dorsal side. P arallelostrombidium ellipticum sp. nov. is recognizable by the anterior ends of the girdle and ventral kineties being close to each other and the posterior ends of the girdle and ventral kineties intersecting on the dorsal side. S trombidium tropicum sp. nov. is distinguished by a ventrally opened girdle kinety that is slightly spiralled with the right end shifted posteriad. Small subunit rRNA gene trees show that P . obesum sp. nov. and P . ellipticum sp. nov. fall into a mixed group composed of Parallelostrombidium and some Novistrombidium species, and that S . tropicum sp. nov. branches at the base of the clade containing non‐Strombidium species. The relationships of Parallelostrombidium species and that of Strombidium species are both not resolved considering their low support values in our phylogenetic analysis. © 2015 The Linnean Society of London     

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.     

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     

Establishment of a new amphileptid genus, Apoamphileptus nov. gen. (Ciliophora, Litostomatea, Pleurostomatida), with description of a new marine species, Apoamphileptus robertsi nov. spec. from Qingdao, China

A new pleurostomatid genus Apoamphileptus is described, which is diagnosed as: Belonging to the Amphileptidae with spica on right side; on each side of the cell, a single perioral kinety, which encircles the cytostome and does not extend to the posterior end of the cell; somatic kineties of both sides near ventral margin shortened and forming a postoral suture; two to several extra fragments with densely arranged dikinetids located in anterior portion of left side. As the type species, the morphology and infraciliature of Apoamphileptus robertsi nov. spec., isolated from a shrimp-farming pond near Qingdao (Tsingtao), China, have been investigated using living observations and the protargol silver impregnation method. The diagnosis for this new species is: Apoamphileptus 90-180 x 30-60 microm in vivo, body elongate pyriform-shaped and slightly flattened; with one cross-striated band along the cytostome; 2-6 (generally 4) large macronuclear nodules, one micronucleus; 33-43 right somatic kineties; left side 6-8 kineties; two extra anterior fragments on left side; about 13 contractile vacuoles dispersed throughout whole body; extrusomes absent or not recognizable; marine habitat. Some morphologically related morphotypes are discussed and tabulated. Regarding the pattern of infraciliature and other morphological features, the well-described fresh-water species, Amphileptus claparedii Stein, 1867 is believed to be a member of this new genus, hence a new combination is suggested: Apoamphileptus claparedii (Stein, 1867) nov. comb.     

Morphology,morphogenesis and molecular phylogeny of a freshwater ciliate,Monomicrocaryon euglenivorum euglenivorum (Ciliophora,Oxytrichidae)

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.     

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.     

Updating the Trachelocercids (Ciliophora, Karyorelictea). I. A Detailed Description of the Infraciliature of Trachelolophos gigas N. G., N. Sp. and T. filum (Dragesco & Dragesco-Kernéis, 1986) N. Comb

ABSTRACT. Trachelolophos gigas n. g., n. sp. and T. filum (Dragesco & Dragesco-Kernéis, 1986) n. comb. (basionym: Tracheloraphis filum) were discovered in the mesopsammon of the French Atlantic coast at Roscoff. Their morphology and infraciliature were studied in live and protargol impregnated specimens. The new genus, Trachelolophos, belongs to the family Trachelocercidae and is unique in having a conspicuous ciliary tuft, which is very likely a highly modified brosse, in the oral cavity. The two species investigated have a very similar infraciliature, differing only in morphometric characteristics and in the nuclear configuration. The entire somatic and oral infraciliature consists of dikinetids which have both basal bodies ciliated or only the anterior or posterior ones, depending on the region of the cell. The right side is densely and uniformly ciliated. Its kineties extend onto the left side to the glabrous stripe, where an anterior and posterior secant system are formed, reducing the number of kineties in the narrowed neck and tail region. The left side bears a narrow glabrous stripe bordered by slightly irregularly arranged dikinetids having rather stiff cilia (bristles), possibly forming an uninterrupted, prolate ellipsoidal (bristle) kinety as indicated by their ciliation. The bristle kinety commences subapically at the right margin of the glabrous stripe, extends posteriorly, then anteriorly at the left, to end up at the right margin again. The dikinetids of the right posterior portion of the bristle kinety have the posterior basal bodies ciliated, whereas the anterior basal bodies are ciliated in its left and right anterior portion. The ends of the bristle kinety meet distinctly subapically at the right margin of the glabrous stripe, as indicated by the diametrically opposed ciliation of the dikinetids. The anterior region (head) of the cell bears a distinct circumoral kinety composed of very regularly arranged dikinetids, associated with nematodesmata forming an oral basket together with the nematodesmal bundles originating from the oralized somatic dikinetids at the anterior end of the somatic kineties. The systematics of trachelocercid ciliates are briefly reviewed and discussed.     

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.     

Morphological Characterizations of Four Species of Parallelostrombidium (Ciliophora,Oligotrichia), with a Note on the Phylogeny of the Genus

The morphology and phylogeny of four oligotrichid ciliates, Parallelostrombidium paraellipticum sp. n., P. dragescoi sp. n., P. jankowskii (Xu et al. 2009) comb. n., and P. kahli (Xu et al. 2009) comb. n., are described or redescribed based on live observation, protargol stained material, and SSU rRNA gene sequences. The new species P. paraellipticum sp. n. is characterized by its obovoidal cell shape, adoral zone composed of 17–21 collar, 9–11 buccal, and two thigmotactic membranelles, and extrusomes attached in one row along the girdle kinety. The new species P. dragescoi sp. n. is distinguished from its congeners by its obovoidal cell shape and a lack of thigmotactic membranelles. Based on ciliary patterns recognizable in the original slides, Omegastrombidium jankowskii Xu et al. 2009 and O. kahli Xu et al. 2009 should be transferred to the genus Parallelostrombidium Agatha 2004. Phylogenetic analyses based on SSU rRNA gene sequence data demonstrate that all four new sequences cluster with previously described congeners. The genus Parallelostrombidium is separated into two clusters, suggesting its non‐monophyly and probably corresponding to the two subgenera proposed by Agatha and Strüder‐Kypke (2014), as well as their morphological difference (cell dorsoventrally flattened vs. unflattened).     

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.     

Morphology of Three Species of Amphileptus (Protozoa,Ciliophora, Pleurostomatida) from the South China Sea,with Note on Phylogeny of A. dragescoi sp. n.

Two new and one problematic species of pleurostomatids, Amphileptus dragescoi sp. n., A. wilberti sp. n., and A. marinus from coastal areas of the South China Sea, are described based on observations of live and protargol‐impregnated specimens. Amphileptus dragescoi is different from its congeners by the presence of an apical group of extrusomes and the possession of 12–15 right and five left somatic kineties, two macronuclear nodules, and a single terminally positioned contractile vacuole. Amphileptus wilberti is diagnosed by oval or pyriform body, 15–19 right and seven or eight left somatic kineties, extrusomes arranged only in anterior portion of oral slit, usually three ventrally located contractile vacuoles, and two macronuclear nodules. Amphileptus marinus (Kahl, 1931) Song et al., 2004 is redescribed and its diagnosis is improved. One isolate which was misidentified as A. marinus by Song et al. (2004) is believed to represent an unknown form, named here as Amphileptus songi sp. n. Phylogenetic analyses of the SSU rDNA sequences indicate that the genus Amphileptus is paraphyletic, but its monophyly is not rejected by statistical tree topology tests.     

Morphology and Phylogeny of Two Novel Pleurostomatids (Ciliophora,Litostomatea), Establishing a New Genus

Pleurostomatida Schewiakoff, 1896 is a cosmopolitan order of ciliates. In the present study, we investigated two new pleurostomatid species, Apolitonotus lynni gen. et sp. nov. and Protolitonotus clampi sp. nov., with state‐of‐the‐art methods. Apolitonotus lynni lacks its oral extrusomes and its right kineties form an anterior semi‐suture near the dorsal margin. Based on these two features, the new genus Apolitonotus was established within the Protolitonotidae Wu et al., 2017. Protolitonotus clampi differs from its congeners by its size of 80–130 × 15–30 μm, 4–6 left, and 9–11 right kineties, extrusomes arranged along the oral slit, and two macronuclear nodules. Because Litonotus antarcticus possesses an anterior semi‐suture and oral extrusomes, it was transferred to the genus Protolitonotus, becoming P. antarctius comb. nov. (basionym Litonotus antarcticus Song and Wilbert, 2002). Phylogenetic analyses based on SSU rDNA sequences suggest a sister group relationship of P. clampi and the family Kentrophyllidae, and A. lynni is adelphotaxon to Litonotus gracilis, both within the order Pleurostomatida. Based on the new findings, an improved diagnosis for Protolitonotus was also provided.     

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 Morphogenesis of Two Species of the Genus Lembadion (Ciliophora, Oligohymenophora): Lembadion lucens and Lembadion bullinum

ABSTRACT. The morphology and morphogenesis of two species of the genus Lembadion, L. lucens and L. bullinum , are described. In both species, left and right ventral kineties converge behind the mouth forming a postoral suture. Buccal infraciliature is formed by one polykinety and two very close paroral kineties (inner and outer). During stomatogenesis, the new oral structures originate from the paroral kineties. The inner paroral kinety forms the new adoral polykinety and regenerates the outer paroral kinety of the proter, while the paroral kineties of the opisthe originate from the outer paroral kinety of the parental cell. Somatic proliferation starts before the stomatogenesis at the equatorial level of the cell, and extends towards the poles forming an equatorial band. Two large invariant zones, anterior and posterior, remain in the dividing cell. Moreover, the kinetodesmal fibers disappear in the proliferation band during the bipartition (fission) process.