Morphology and Molecular Phylogeny of Two New Ciliates,Holostichides heterotypicus n. sp. and Holosticha muuiensis n. sp. (Ciliophora: Urostylida)

Two new urostylid species, Holostichides heterotypicus n. sp. and Holosticha muuiensis n. sp., were discovered in South Korea. Morphological and phylogenetic analyses were carried out to confirm that these species are new to science. Holostichides heterotypicus is mainly characterized by the following combination of features: 110–205 μm long in vivo; 5–10 frontoterminal cirri; 6–8 midventral pairs with 2–3 midventral cirral rows; cortical granules present; four bipolar dorsal kineties; and 6–9 caudal cirri. Ontogenetic features of H. heterotypicus are similar to those of H. typicus. Phylogenetic analyses revealed that H. heterotypicus was distantly separated from bakuellid genera Apobakuella, Bakuella, Metaurostylopsis, and Neobakuella. This result is supported by the following features: transverse cirri (present in the other four bakuellids vs. absent in Holostichides) and caudal cirri (absent in the other four bakuellids vs. present in Holostichides). Holosticha muuiensis n. sp. is mainly distinguished from its congeners by the following combination of features: 100–185 long in vivo; shortened undulating membrane; cortical granules lacking; contractile vacuole absent; 51–66 adoral zone of membranelles; 42–60 macronuclear nodules; and five bipolar dorsal kineties. In the phylogenetic tree, Holosticha muuiensis n. sp. clustered with a Holosticha group (containing Holosticha diademata, Holosticha foissneri, and Holosticha heterofoissneri).     

Redescription of two little-known urostyloid ciliates,Anteholosticha randani () and A. antecirrata (Ciliophora,Urostylida)

The morphology of two little-known urostyloid ciliates, Anteholosticha randani (Grolière, 1975) Berger, 2003 and A. antecirrata Berger, 2006, collected from freshwater biotopes in southern China, was studied based on live observations and protargol staining. Anteholosticha randani is characterized by its bipartite adoral zone and short, longitudinally aligned undulating membranes. One early stage of reorganization/morphogenesis, one early-middle stage of reorganization and one middle stage of morphogenesis are also reported. Anteholosticha antecirrata is characterized by its large body size in vivo (200–400 × 40–80 μm), a row of buccal cirri and conspicuous, yellow-green cortical granules. Phylogenetic analyses based on SSU rDNA sequence data reveal that A. antecirrata may share a most recent common ancestor with Urostyla grandis and Bakuella granulifera, whereas A. randani branches independently and is sister to a large clade that includes Pseudourostyla, Pseudokeronopsis, Caudiholosticha and several species of Anteholosticha.     

A Comparative Analysis of the Morphology of Holosticha corlissi N. Sp. (Ciliophora,Hypotrichida) and Related Species1

ABSTRACT The morphology of the hypotrichous ciliate, Holosticha corlissi n. sp., found in the moss Calliergonella cuspidata taken from the beech wood of Montejo de la Sierra (Madrid, Spain) is described. Some characteristics (organization of the mid-ventral, frontal and fronto-terminal cirri, presence of a buccal cirrus and the number of transverse cirri) are sufficiently different from the closely related species H. intermedia, H. sigmoidea and H. xanthichroma to suggest that it is a separate species, although its body shape, nucleus and buccal apparatus are very similar.     

An Analysis of the Formation of Ciliary Primordia in the Hypotrich Ciliate Urostyla weissei*

SYNOPSIS. The protargol technic was used in a study of the development of oral, cirral, and dorsal primordia of Urostyla weissei fixed during division, reorganization, and regeneration following transection at different levels. While the course of development is similar in all situations, differences were observed in the way in which some primordia are initiaily formed. The primordium of the new AZM always appears posterior to the old AZM. It develops into an entire new membranellar band in dividing cells and in opimers (posterior fragments from equatorial transections), while it eventually joins with a portion of the old AZM in reorganizers, promers (anterior fragments from equatorial transections) and “large opimers” (cells whose anterior tip has been cut off). The UM-primordium of proters is derived from disaggregation of the kinetosomes of the 2 old UM's, that of opisthes and opimers is formed “de novo” to the right of the AZM-primordium, while the UM-primordium of reorganizers, promers, and “large opimers” is of composite origin, partly “de novo” and partly from the old UM's. The UM primordium differentiates into the new UM's and the 1st frontal cirrus. The primordia of the remaining frontal, ventral, transversal (F-V-T) and marginal cirri originate as “streaks” of cilia, most of which are derived from re-alignment of the constituent cilia of certain pre-existing cirri. New cirri differendiate from the streaks, and replace the remaining old cirri. The streaks are formed similarly in all developmental situations, except for the 1st 3 F-V-T streaks. In proters, reorganizers, and promers, these originate from the posterior 3 frontal cirri, while in opisthes and opimers they are formed “de novo” to the right of the UM-primordium. In the “large opimers” these streaks are formed “de novo” behind the 1st 3 frontal cirri, in spite of the continued presence of these cirri at the anterior tip of the fragments. The site of formation of these streaks thus appears to be determined by an anteriorposterior gradient, rather than by any preformed cortical structure. The new dorsal bristle rows I to III develop from the proliferation of portions of the old rows, while rows IV and V originate from short kineties formed “de novo” on the right margin. New caudal cirri differentiate at the posterior ends of the new rows I to III. The numbers of ventral cirral rows and transversal cirri are variable; these variations are correlated, and related to variations in numbers of developing streaks. A survey of hypotrich developmental patterns revealed extensive parallels, especially in the sites of appearance of primordia. The primordium site appears to be a more constant feature of cortical development than is the “source” of ciliary units. It is concluded that sites of primordia are determined by cellular gradients, with competent preformed structures being utilized if they are appropriately positioned within these gradients.     

Palp morphology in two species of Prionospio (Polychaeta: Spionidae)

The palp morphology of Prionospio fallax Söderström, 1920 from Sweden and Prionospio cf. saldanha Day, 1961 from Thailand was examined with a scanning electron microscope. Prionospio fallax was also studied in vivo using light-microscopy. Both species have grooved feeding palps, adorned with up to five ciliary characters: frontal cilia, transverse ciliary bands (or bandlets), latero-frontal cirri, lateral cilia and randomly distributed non-motile cirri. All, except the frontal cilia and non-motile cirri, are asymmetrically arranged relative to the long axis of the palps. Prionospio fallax possesses transverse bandlets and the other four groups, while P. cf. saldanha has transverse bands (consisting of several contiguous bandlets), frontal cilia and some randomly scattered cirri. Asymmetrical palp ciliation was previously only known in Marenzelleria viridis (Verril, 1873) and the genus Scolelepis Blainville, 1828. The newly recognised transverse ciliary bands and bandlets are considered to be homologous with the transverse ciliary rows found basally on the palps of Paraprionospio pinnata (Ehlers, 1901). This multistate character (named transverse cilia) may prove useful in elucidating the phylogeny of the Prionospio-complex of genera.     

Morphology and Small Subunit rDNA Phylogeny of Two New Marine Urostylid Ciliates,Caudiholosticha marina sp. nov. and Nothoholosticha flava sp. nov. (Ciliophora,Hypotrichia)

Two marine urostylid ciliates, Caudiholosticha marina sp. nov. and Nothoholosticha flava sp. nov., isolated from intertidal sediment in the Yellow Sea, are investigated using morphological and small subunit rDNA phylogenetic analyses. Caudiholosticha marina is 210?310 μm × 40?55 μm in vivo, and has 10?20 macronuclear nodules, 23?37 midventral cirral pairs extending to 5?8 transverse cirri, and two caudal cirri. It differs from congeners by its marine habitat, larger size, macronuclear arrangement pattern and high number of midventral pairs. Molecular phylogenetic analyses indicate a polyphyly of Caudiholosticha. Nothoholosticha flava is yellow to brownish and 240?320 μm × 40?60 μm sized, and has a bipartite adoral zone, six frontal cirri in atypical bicorona, usually four frontoterminal, one buccal and 5?7 transverse cirri and 28?54 midventral pairs. Phylogenetic analyses allocate N. flava as sister of N. fasciola, type of the genus. The two Nothoholosticha species differ distinctly by the presence/absence of frontoterminal cirri, a feature often used to define genera in the Hypotrichia. However, the SSU rDNA sequence similarity between these two species is 99.3%, which weakens the justification for separating the new isolate at genus level. The taxonomic significance of frontoterminal cirri is discussed based on morphological and molecular data.     

Molecular Phylogeny and Taxonomy of Two Novel Brackish Water Hypotrich Ciliates,with the Establishment of a New Genus,Antiokeronopsis gen. n. (Ciliophora,Hypotrichia)

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.     

The Structure and Morphogenesis of the Ventral Ciliature in Paraurostyla hymenophora*

SYNOPSIS. The structure and morphogenesis of the ventral ciliature of Paraurostyla hymenophora (Stokes) are described. The oral primordium apparently originates in association with transverse cirrus #6, from which it migrates anteriorly simultaneous with kinetosomal proliferation. The primordium eventually forms an elongate ciliary field from which the future opisthe's fronto-ventro-transverse (FVT) and undulating membrane primordial fields arise. Concomitantly, the future proter's FVT primordial field is initiated by the disaggregation of frontal cirri #4, #5, and #6. Primordia then develop simultaneously within marginal and ventral cirral rows by a disaggregation of cirri within the respective rows, and do not give rise to new cirri until the FVT fields complete segregation into discrete cirri. Near the completion of cirral production from the FVT primordia, each ventral cirral primordium (VCP) forms the 2 rightmost transverse cirri. Segregation of new cirri within the marginal cirral primordia and VCP then occurs, eventually replacing all old cirri within their respective marginal and ventral cirral rows. At the end of cortical morphogenesis, all old ciliary organelles, with the exception of the adoral zone of membranelles, are either reorganized or replaced. These results suggest an evolutionary affinity between the ventral and marginal cirral rows and raise questions about the control of the developmental competence of individual primordia.     

Pattern Formation in Mirror-Image Doublets of the Ciliate Stylonychia pustulata

ABSTRACT. Mirror-image symmetry doublets of the ciliate Stylonychia pustulata were obtained from the progenies of dividing cells in which cell division was inhibited by heat-shocks. In two components consisting of the doublet, the left (cell's) component possessed ciliary organelles arranged in almost the same pattern as in normal singlets, while the right one had surface organelles located in a mirror-image symmetry of those of the left component. In cell division of the doublet, two sets of ciliary primordia that were arranged in a mirror-image symmetry developed synchronously in both components. In about 80% of oral primordia (OP) of the right components, the arrangement of the membranellar bands became abnormal. In some cases, OP of the right component were occasionally separated into two longitudinal halves, each consisting of normal membranelles and inverted membranelles. A set of primordia of the paroral membranelles and fronto-ventro-transverse cirri was rarely derived from the basal bodies of the right half with a band of normal membranelles. As a result, a third component with the ciliary organelles normally arranged emerged on the right side of the original right component. The differentiation of membranelles and segmentation of the primordial streaks into cim proceeded from anterior to posterior. A cytoplasmic bulge with multiple right marginal cirral rows was frequently formed at the right margin of the doublet. The behavior in the separation of third and fourth streaks from a primordial streak of dorsal cirri was not mirror-image symmetrical in each component.     

Description of the Halophile Euplotes qatarensis nov. spec. (Ciliophora,Spirotrichea, Euplotida) Isolated from the Hypersaline Khor Al‐Adaid Lagoon in Qatar

The morphology, ontogenesis, and phylogenetic relationships of a halophile euplotid ciliates, Euplotes qatarensis nov. spec., isolated from the Khor Al‐Adaid Lagoon in Qatar were investigated based on live observation as well as protargol‐ and silver nitrate‐impregnated methods. The new species is characterised by a combination of features: the halophile habitat, a cell size of 50–65 × 33–40 μm, seven dorsal ridges, 10 commonly sized frontoventral cirri, two widely spaced marginal cirri, 10 dorsolateral kineties, and a double silverline pattern. The morphogenesis is similar to that of its congeners: (i) the oral primordium develops hypoapokinetally and the parental oral apparatus is retained; (ii) the frontoventral‐transverse field of five streaks gives rise to the frontal, ventral, and transverse cirri, but not to the cirri I/1 and the marginal cirri; (iii) the dorsal somatic ciliature develops by intrakinetal proliferation of basal bodies in two anlagen per kinety that are just anterior and posterior to the future division furrow; (iv) the caudal cirri are formed by the two rightmost dorsolateral kineties. The SSU rDNA sequence of E. qatarensis branches with full support in the Euplotopsis elegans–Euplotes nobilii–Euplotopsis raikovi clade. The closest related publicly available SSU rDNA sequence is the one of E. nobilii, with which E. qatarensis has 93.4% sequence similarity. Euplotes parawoodruffi Song & Bradbury, 1997 is transferred to the genus Euplotoides based on the absence of frontoventral cirrus VI/3.     

A redescription of the marine hypotrichous ciliate, Nothoholosticha fasciola (Kahl, 1932) nov. gen., nov. comb. (Ciliophora: Urostylida) with brief notes on its cellular reorganization and SS rRNA gene sequence

The morphology and infraciliature of the marine hypotrichous ciliate Nothoholosticha fasciola (Kahl, 1932) nov. gen., nov. comb., isolated from mariculture waters near Qingdao, China, are redescribed based on live and protargol-impregnated specimens. Features reported for the first time include the possession of more than 50 macronuclear nodules and details of the infraciliature, i.e. 50–60 adoral membranelles, shortened paroral and endoral membranes, six frontal, one buccal and two to seven transverse cirri, ca. 40 pairs of midventral cirri, ca. 60–120 left and 70–120 right marginal cirri, three dorsal kineties, caudal and frontoterminal cirri absent. In addition, brief details of two stages of cellular reorganization in N. fasciola are supplied and comparisons with some related urostylids based on SS rRNA gene sequence data are reported. The new genus Nothoholosticha is established based primarily on the absence of frontoterminal cirri, which distinctly separates it from similar urostylid genera. Anteholosticha longissima is transferred to Nothoholosticha as N. longissima (Dragesco and Dragesco-Kernéis, 1986) nov. comb. and Holosticha antarctica is transferred to Pseudokeronopsis as P. antarctica (Wilbert and Song, 2008) nov. comb.     

Morphology and Molecular Phylogeny of a New Hypotrich Ciliate,Pseudourostyla guizhouensis sp. nov. from Southern China,with Notes on a Chinese Population of Hemicycliostyla franzi (Foissner, 1987) Paiva et al., 2012 (Ciliophora,Hypotricha)

The morphology and molecular phylogeny of a soil hypotrich ciliate, Pseudourostyla guizhouensis sp. nov., collected from southern China, were investigated. Pseudourostyla guizhouensis sp. nov. has an elongate elliptical body measuring 180–310 × 65–85 μm in vivo; invariably two right and three or four left marginal rows; six or seven dorsal kineties; adoral zone consisting of 57–70 membranelles; 12–16 frontal cirri, one buccal cirrus, 13–20 midventral pairs, two frontoterminal cirri, two pretransverse cirri, and five to seven transverse cirri. Morphogenesis during physiological regeneration indicates that the marginal rows of each side originate from a common anlage that differentiates into several rows. Molecular phylogenetic analysis based on SSU rDNA sequence data reveals that P. guizhouensis sp. nov. clusters with the type species P. cristata (Jerka‐Dziadosz, 1964) Borror, 1972 and that the genus Pseudourostyla is monophyletic. The morphological characters of another soil hypotrich ciliate, Hemicycliostyla franzi (Foissner, 1987) Paiva et al., 2012, are also described based on a Chinese (Guizhou) population.     

Effects of calcium on the longituindal flagellum of Oxyrrhis marina

Summary— 2–4 nm filaments represent a new class of cytoskeletal components. They are found in ciliary and flagellar roots and centrosomes of all eucaryotes. They are also the major components of paraflagellar rods (PFR) in Euglena, trypanosomes and dinoflagellates. Oxyrrhis marina, a marine dinoflagellate, possesses a transverse and a longitudinal flagellum. Only the longitudinal flagellum carries the PFR along the proximal two-thirds of its length. This flagellum is not only capable of the classic flagellar beat but is also able to retract and bend, a property mediated by external calcium. To determine if calcium has a direct role in the bending, experimental conditions were established to permeabilization and reactivation. Our conditions to reactivate the axoneme function (wave propagation) appear similar to those observed in the case of the sea urchin sperm. The results show that in vitro, an increase in calcium concentration induces a conformational change of the longitudinal flagellum in the absence of ATP with a half maximum effect at 0.1 μM. In the presence of ATP, this morphology modification causes a total inhibition of the wave propagation which is replaced by non-propulsive contractions of low amplitude. As these properties are not shared by reactivated sea urchin sperm flagella or the transverse flagellum of O marina devoid of PFR, we propose that PFR are responsible for the bending phenomenon. A calcium shock also induces flagellar excision with a half maximum effect at 0.3 μM, and immunofluorescence results suggest that a centrin-like protein is present in O marina and is responsible for this excision.     

Cladotricha and Phylogeny in the Suborder Stichotrichina (Ciliophora,Hypotrichida)*

Synopsis. The cortical anatomy and morphogenesis of Cladotricha koltzowii Gajewskaja and Cladotricha variabilis Rumen from the Great Salt Lake, Utah, are described. When compared with 2 marine hypotrichs, Uroleptoides kihni Wenzel and Stichotricha secunda Kahl, from the east coast of the United States, their structure and development suggest that Cladotricha may have descended from an intermediate species within the line of evolution of the Spirofilidae from primitive Kahliella-like hypotrichs. It is suggested that the Kahliella-Cladotricha-Uroleptoides series is separated at the familial level from both the tubicolous and planktonic spirofilids. Further, the structural and morphogenetic differences between Cladotricha and the holostichid Uroleptus appear to be at a subordinal level. These findings are significant in revising definitions of the Hypotrichida, its suborders, and the Urostylidae.     

Schmidingerothrix salinarum nov. spec. is the Molecular Sister of the Large Oxytrichid Clade (Ciliophora,Hypotricha)

In 2012, Foissner described a curious hypotrich: Schmidingerothrix extraordinaria. This ciliate, which he discovered in hypersaline soils (~100‰) from Namibia, had a frayed buccal lip, three‐rowed adoral membranelles, only one frontal cirrus, and a miniaturized first frontal membranelle, while a paroral membrane, dorsal bristle rows and buccal, transverse and caudal cirri were absent. All opisthe structures developed de novo, while parental structures were involved in the proter. When Foissner's study became available, we discovered a similar species in a Portuguese solar saltern, differing from S. extraordinaria mainly by the number of frontoventral cirral rows (3 vs. 1). Furthermore, parental structures were involved in the ontogenesis of both proter and opisthe. The small subunit (SSU) rDNA shows Schmidingerothrix as sister of a large clade containing most classical oxytrichids (e.g. Sterkiella, Oxytricha, Steinia) and many related taxa (e.g. Pattersoniella, Bistichella, Uroleptus). This clade shows a bifurcation named “Oxytricha subclade” and “Uroleptus subclade”. Foissner ( 2012 ) interpreted the peculiarities of Schmidingerothrix as a reduction caused by the extreme habitat. However, the molecular data do not exclude that Schmidingerothrix presents an ancient state. A morphology‐based scheme is presented, showing how the subclades might have evolved from a Schmidingerothrix‐like ancestor.     

Morphology and infraciliature of Holosticha bradburyae nov. spec. (Ciliophora,Hypotrichida) from the Yellow Sea,China

The living morphology and infraciliature of a new marine hypotrichous ciliate, Holosticha bradburyae nov. spec., collected from the coastal water off Qingdao (Tsingtao), China, are investigated. This species is characterized by: body size 150–320 × 25–75 m with brownish to dark brown cell colour, ca. 53 adoral membranelles and 1 anteriorly positioned buccal cirrus; 3 frontal, 2 frontoterminal and 20–26 transverse cirri; midventral rows comprising 27-32 pairs of cirri; one conspicuous gap always present between anterior and posterior parts of AZM, and 2–5 distinctly elongated membranelles are always present at the posteriormost end; cortical granules conspicuous, round and flattened with central depression, arranged in about 10 lines on dorsal side; 28–33 irregularly arranged macronuclear nodules; 9–11 complete dorsal kineties; caudal cirri absent.     

Morphology,ontogeny and molecular phylogeny of a new urostylid ciliate,Bakuella (Pseudobakuella) guangdongica n. sp. (Ciliophora,Hypotrichia) from southern China

The morphology, morphogenesis, and molecular phylogeny of Bakuella (Pseudobakuella) guangdongica n. sp., isolated from southern China, were investigated. The new species is characterized by a body length of 150–225 μm in vivo; 35–42 adoral membranelles; 3–5 buccal, two frontoterminal, 7–12 transverse and two pretransverse ventral cirri; midventral complex comprised of 10–20 pairs and two rows extending to transverse cirri; posterior part of marginal rows slightly overlapping; colorless cortical granules about 1 μm across, arranged in small groups; soil habitat. Its main ontogenetic features are: (1) in the proter, the parental adoral zone of membranelles is completely renewed by new structures; (2) in the opisthe, the oral primordium originates apokinetally, some old midventral cirri join the formation of frontoventral-transverse cirral anlagen; (3) the anlagen for marginal rows and dorsal kineties develop intrakinetally; and (4) the numerous macronuclear nodules fuse into a single mass before dividing. Phylogenetic analyses based on the SSU rDNA sequence suggest the non-monophyly of the genus Bakuella.     

Morphology,morphogenesis and molecular phylogeny of a new marine ciliate,Trichototaxis marina n. sp. (Ciliophora,Urostylida)

The live morphology, infraciliature and morphogenesis of a new urostylid ciliate, Trichototaxis marina n. sp., collected from coastal water in Qingdao, China, were studied based on the observations of live and silver stained specimens. The new species is characterised as follows: body very flexible and contractile, slight to brick-reddish in colour due to irregularly-shaped, brick-red pigments; ca. 70 adoral membranelles; about 17 frontal cirri arranged in a bicorona; average 67 midventral pairs, the right base of each pair being conspicuously larger than the left base; five to seven transverse cirri; constantly two frontoterminal, one buccal and two pretransverse ventral cirri; two or three left marginal rows; right and innermost left marginal rows with 56–92 and 66–106 cirri, respectively; six bipolar dorsal kineties; more than 100 macronuclear nodules. The characteristic morphogenetic feature in T. marina is the development of the left marginal rows, that is, only one left marginal row is newly built the other one or two being retained from the parental cell. Phylogenetic analyses based on small subunit ribosomal gene sequence data reveal a close relationship of T. marina with members of family Pseudokeronopsidae.