Morphology and infraciliature of two new marine urostylid ciliates: Metaurostylopsis struederkypkeae n. sp. and Thigmokeronopsis stoecki n. sp. (Ciliophora, Hypotrichida) from China

ABSTRACT. The morphology and infraciliature of two new marine urostylid ciliates, Metaurostylopsis struederkypkeae n. sp. and Thigmokeronopsis stoecki n. sp., collected from the coastal waters off Qingdao (Tsingtao), China, have been investigated. Metaurostylopsis struederkypkeae n. sp. is characterized by the slender body shape, small size, rose-reddish cell colour, and having two kinds of pigment-like granules. The larger pigment-like granules are yellow–green or grass-green in colour, oval in shape, and flattened, whereas the smaller ones are wine-reddish. Infraciliature and nuclear apparatus are similar to the well-known Metaurostylopsis marina. Thigmokeronopsis stoecki n. sp. is characterized by its large size with dark brown cell colour and grass-green cortical granules, which are large, blood-cell shaped, and sparsely distributed. The thigmotactic ciliature is conspicuous: 11–14 rows of densely arranged cirri occupy the most postoral area. Keys are provided for all the known species in both genera.     

Morphology and Molecular Phylogeny of Two New Marine Euplotids,Pseudodiophrys nigricans n. g., n. sp., and Paradiophrys zhangi n. sp. (Ciliophora: Euplotida)

ABSTRACT. The morphology, infraciliature, and molecular phylogeny of Pseudodiophrys nigricans n. g., n. sp., and Paradiophrys zhangi n. sp., isolated from a sandy beach near Qingdao, China, were investigated. Pseudodiophrys is characterized by the Diophrys‐like ciliature pattern, but having only a single, reduced undulating membrane. Pseudodiophrys nigricans, the type species by monotypy, is described from live and silver‐impregnated specimens. Paradiophrys zhangi is similar to the type species Paradiophrys irmgard but can be recognized by its border body and the number (7 vs. 8–10) and arrangement (in rows vs. sparsely distributed) of the frontoventral cirri. Small subunit (SSU) rRNA gene sequence data support the validity of both species. Phylogenetic analyses based on the SSU rRNA gene sequence data currently available for uronychiids and other related taxa indicate that P. nigricans is most closely related to Diophrys scutum and Diophrys apoligothrix, while P. zhangi clusters most closely with Apodiophrys ovalis within a clade that also includes two other Paradiophrys species.     

Kiitricha minuta n. sp., a peculiar hypotrichous ciliate (Ciliophora, Spirotrichea) from the Yellow Sea

The morphology and infraciliature of a new ciliate, Kiitricha minuta n. sp., isolated from the Yellow Sea, were investigated using live observation and protargol impregnation. Kiitricha minuta represents a third member of the rarely known order Kiitrichida. It is unique in the subclass Hypotrichia in having many rows of small uniform cirri along the right side of the body and the dorsal kineties composed of dikinetids, most of which bear two cilia. Kiitricha minuta n. sp. is ovoid and measures about 60 x 45 microm in vivo. It has a huge buccal cavity occupying about 80% of the body length, numerous body extrusomes, one macronucleus and two micronuclei, 27-27 adoral membranelles, 9-12 frontoventral cirral rows, a submarginal row of 7-9 cirri, 6 or 7 transverse cirri, and roughly 7-9 dorsal kineties. This new species differs distinctly from its only congener Kiitricha marina by its smaller size (60 microm vs. 80-150 microm), the presence of body extrusomes (vs. absent), the different macronuclear pattern (one vs. two nodules), and the lower number of frontoventral cirral rows (9-12 vs. 21-26), which terminate at the anterior two-thirds of body (vs. extend to the posterior). The new term "submarginal cirral row" is introduced to distinguish from the marginal cirral row in typical hypotrichs sensu lato. Based on our new observations and the literature, an improved diagnosis for the genus Kiitricha is provided and its phylogenetic importance is discussed.     

Ontogenesis and molecular phylogeny of a new marine urostylid ciliate, Anteholosticha petzi n. sp. (Ciliophora, Urostylida)

A new urostylid ciliate, Anteholosticha petzi n. sp., isolated from the northern China seas, was studied using live observation and protargol impregnation. It differs from all the congeners mainly in the highly flexible and variable body shape, its red-brown to brick-red colour, and the possession of three types of cortical granules. Analysis of morphometric and molecular data confirm the validity of A. petzi as a distinct species. Anteholosticha petzi n. sp. is characterized as follows: by having 3 frontal, 1 buccal, 2 frontoterminal, 2 pretransverse, and 8-11 transverse cirri; a midventral row that comprises 10-16 cirral pairs and extends ca. 60% of cell length; and 3 bipolar dorsal kineties. Ontogenesis is similar to that in Anteholosticha warreni except for the formation of the oral primordium and is characterized by: (1) the parental adoral membranelles are completely renewed and the oral primordium is formed de novo; and (2) the anlagen for the frontal-ventral-transverse cirri are formed as primary primordia. The small subunit rRNA gene sequence of A. petzi n. sp. was also compared with that of its congeners with the sequence differences ranging from 0.39% to 8.62%. Phylogenetic trees based on the SSU rRNA gene sequence were constructed, indicating the nonmonophyly of the genus Anteholosticha, as supported by the approximately unbiased test.     

Tunicothrix rostrata n. g., n. sp., a new urostylid ciliate (Ciliophora, Stichotrichia) from the Yellow Sea

The morphology and infraciliature of a new ciliate, Tunicothrix rostrata n. g., n. sp., isolated from the Yellow Sea, are investigated using live observation and protargol impregnation. Tunicothrix rostrata measures about 160 x 40 mum in vivo, and has a frontal beak-like protrusion, a conspicuous cortical alveolar layer, two right marginal rows, and usually three distinct midventral pairs. The discovery of T. rostrata enables us to reconsider the classification of Erniella wilberti, a curious ciliate with obscure midventral pairs. Both species are highly similar in overall appearance and nuclear and ciliary pattern. Thus, they are united in a new genus of the family Urostylidae, Tunicothrix, and E. wilberti is transferred to Tunicothrix: Tunicothrix wilbertiLin and Song, 2004 n. comb. By contrast, Erniella filiformis, type species of Erniella, has several ventral rows and does not belong to the urostylids. Tunicothrix rostrata is easily distinguished from T. wilberti by its beak-like anterior protrusion and by the distinctly elongated right marginal row 2, which curves anteriorly on the dorsal side of the cells. Tunicothrix is closely related to Parabirojimia, differing by the invariably two (vs. five-eight) right marginal rows and the conspicuous (vs. ordinary) alveolar layer, a unique feature in urostylid ciliates.     

Morphology and infraciliature of three species of Metaurostylopsis (Ciliophora, Stichotrichia): M. songi n. sp., M. salina n. sp., and M. marina (Kahl 1932) from sediments, saline ponds, and coastal waters

Two new urostylid ciliates, Metaurostylopsis songi n. sp. and Metaurostylopsis salina n. sp. and Metaurostylopsis marina (Kahl 1932) are investigated using live observation and protargol impregnation. These species were isolated in Korea from intertidal sediments, saline ponds, and coastal waters. Metaurostylopsis songi is in vivo about 120 microm x 25 microm, has a slenderly ellipsoidal body, colorless cortical granules in rows on ventral and dorsal body sides, about 54 macronuclear nodules, 28-47 adoral membranelles, five frontal, two or three frontoterminal and six or seven transverse cirri, and 9-12 midventral cirral pairs followed posteriorly by 1-3 single cirri. In vivo M. salina is about 60 microm x 25 microm, has a pyriform body, colorless cortical granules irregularly arranged, about 45 macronuclear nodules, 18-23 adoral membranelles, three frontal, three to five frontoterminal and two to five transverse cirri, and four or five midventral cirral pairs followed posteriorly by five to seven single cirri. Both species have three marginal cirral rows on each body side and 3 long dorsal kineties. The Korean specimens of M. marina match the Chinese population in all main features. Metaurostylopsis songi differs from M. marina by the more slender body, the number of frontal cirri (invariably five vs. four), and the arrangement of cortical granules (in rows on dorsal and ventral cortex vs. only along dorsal kineties and anterior body margin). Metaurostylopsis salina differs from its congeners by the distinctly smaller size, the pyriform body shape, the scattered cortical granules (vs. in rows), and number of frontal cirri. It differs from M. marina also by the number of midventral cirral pairs (four or five vs. seven to 11).     

Description of a marine peritrichous ciliate, Pseudovorticella sinensis n. sp. (Ciliophora, Peritrichia) from China

A new marine peritrich ciliate, Pseudovorticella sinensis n. sp. was isolated from a shrimp-farming pond in the littoral area of Qingdao, China. The morphology, infraciliature, and silverline system were studied based on living and silver-impregnated specimens. This species is characterized by (1) an elongated bell-shaped body that measures 50-60 x 35-45 microm in vivo, (2) one large, ventrally located contractile vacuole, and (3) a pellicle covered by a layer of transparent, cortical vesicles. The number of transverse silverlines from the peristomial area to the aboral ciliary wreath is 26-32, and from the aboral ciliary wreath to the scopula is 12-15. The stalk measures about 160-250 microm long x 5-6 microm wide. The spasmoneme has one row of conspicuous thecoplasmic granules, which are about 0.8 microm in diameter.     

Morphology and small subunit rDNA phylogeny of a new soil ciliate, Bistichella variabilis n. sp. (Ciliophora, Stichotrichia)

We isolated a new ciliate, Bistichella variabilis n. sp., from slightly saline soil of the Yellow River Delta in China and studied it using live observation, protargol staining, and small subunit (SSU) rRNA gene sequencing. Bistichella variabilis is elongated ellipsoidal and about 220 × 70 μm in vivo, with a variable number and arrangement of cirri- ventral row VI mostly very short, but a long row observed also in one specimen, and an extra row occasionally exists besides the left or the right marginal row. Bistichella variabilis differs from all congeners except Bistichella buitkampi (Foissner, 1982) in the macronuclear pattern (four vs. two or many nodules); and from B. buitkampi by the usual oblique arrangement of transverse cirri (vs. perpendicular); and the ventral row VI mostly restricted in the posterior quarter of the cell (vs. long and stretching to the anterior quarter). Phylogenetic analyses of SSU rDNA sequences indicated that B. variabilis grouped with the clade of Orthoamphisiella breviseries and Uroleptoides magnigranulosus with high support values. Despite their distinct differences in morphology, these three species have sequence similarities of 99.5-99.7%. Nonetheless, it is still premature to assign Bistichella together with Uroleptoides to the family Orthoamphisiellidae because of the taxonomic uncertainties and undersampling.     

The morphology, ontogeny, and small subunit rRNA gene sequence analysis of Diophrys parappendiculata n. sp. (Protozoa, Ciliophora, Euplotida), a new marine ciliate from coastal waters of southern China

The morphology, morphogenesis, and phylogeny of Diophrys parappendiculata n. sp., a large marine ciliate isolated from the coastal waters of Daya Bay, southern China, were investigated. This new species is characterized by a combination of its large size, appendiculata-pattern of ciliature, and bipartite adoral zone of membranelles. The main stages of morphogenesis during binary fission were also recorded and described. Comparisons of morphological characteristics with similar congeners support the validity of the new species. The small subunit rRNA gene sequence of D. parappendiculata is 96.3-99.94% similar to those of four other congeners; it differs in four nucleotides from that of Diophrys appendiculata (i.e. structural similarity was 99.94%). Phylogenetic analysis indicates that D. parappendiculata n. sp. falls into the Diophrys clade and is most closely related to the well-known D. appendiculata.     

悬游双眉虫(原生动物,纤毛门)无性生殖期间的形态发生

应用蛋白银染色技术研究了悬游双眉虫青岛种群无性生殖期间皮层结构和核器的演化过程,其主要特征为:后仔虫口原基独立地出现于紧靠虫体左侧第一根横棘毛的皮层下小龛,其中毛基粒不参与其它棘毛原基的形成;老的口围带发生后半部的局部重建而非整个的由前仔虫简单继承;在前仔虫中,波动膜原基来自老结构的反分化,而在后仔虫中则来自口原基;所有棘毛原基均为独立发生并与老结构没有任何关系;在前仔虫中,口棘毛(即左侧第一根额棘毛)来自于波动膜的反分化,而在后仔虫中则为独立发生;背触毛列于老的结构当中产生,并由最右一列原基演化出3根尾棘毛;两大核片段的改组带从一端向另一端移动 ,并随着两者的融合而消失.文中同时对前人有关该属发生模式的若干存疑问题做了探讨 [动物学报 54(3):517-524,2008].     

Morphology and phylogeny of a new urostylid ciliate, Monocoronella carnea n. g., n. sp. (Ciliophora, Hypotricha) from Daya Bay, Southern China

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.     

Morphology and Small Subunit rDNA Gene Sequence of Pseudoamphisiella quadrinucleata n. sp. (Ciliophora,Urostylida) from the South China Sea

ABSTRACT. The urosylid genus Pseudoamphisiella was established by Song (1996) with hitherto only two known congeners. In the present work, the morphology and infraciliature of a new member, Pseudoamphisiella quadrinucleata n. sp., a form with conspicuous alveolar layer and four macronuclear nodules isolated from the coastal waters both near Hong Kong and near Guangzhou, South China were investigated using living observation and protargol silver impregnation methods. Pseudoamphisiella quadrinucleata differs from other two known forms mainly by the number of macronuclear nodules: constantly four vs. two in Pseudoamphisiella alveolata and 24–57 in Pseudoamphisiella lacazei. To support this, the sequence of the small subunit rDNA of P. quadrinucleata n. sp. showed 14 and 74 nucleotides in comparison with that of the two known congeners, respectively, which hence firmly supports the validity of the new species.     

Morphology,morphogenesis and molecular phylogeny of a novel soil ciliate,Afrokahliella paramacrostoma n. sp. (Ciliophora,Hypotrichia)

A soil hypotrich ciliate, Afrokahliella paramacrostoma n. sp., was discovered in China. Its morphology, morphogenesis and molecular phylogeny were investigated using standard methods. The new species is characterized as follows: body about 140–180 × 60–70 μm in vivo, cortical granules absent, contractile vacuole positioned about 40% down length of body, 5–9 macronuclear nodules, 34–49 adoral membranelles, 3–5 buccal and 3–6 parabuccal cirri, usually two frontoventral rows, three or four left and two or three right marginal rows, three dorsal kineties and one dorsomarginal kinety; 1–3 and one or two caudal cirri located at the ends of dorsal kineties 1 and 2, respectively. The ontogenetic process is characterized by: (1) the marginal anlagen on each side develop in the outer right and the inner left marginal rows, respectively; (2) five frontoventral-transverse cirral anlagen, anlagen II–IV develop in secondary mode; (3) dorsal morphogenesis follows a typical Urosomoida-pattern, no parental dorsal kineties are retained; (4) caudal cirri are generated at the ends of dorsal kineties 1 and 2. Phylogenetic analyses based on SSU rDNA sequence data reveals that Afrokahliella paramacrostoma n. sp. is closely related to Parakahliella macrostoma and Hemiurosomoida longa.     

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 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.     

Ontogeny and molecular phylogeny of a new marine ciliate genus,Heterokeronopsis g. n. (Protozoa,Ciliophora, Hypotricha), with description of a new species

The morphology and morphogenesis of a new ciliate, Heterokeronopsis pulchra g. n., sp. n., isolated from a mangrove wetland near Shenzhen, southern China, were investigated using live observation and protargol impregnation methods. Heterokeronopsis g. n. is characterized by having a bipartite adoral zone, an extremely shortened paroral membrane, frontal cirri arranged in an indistinct bicorona, a midventral complex composed of midventral pairs and midventral row(s), one left and one right marginal row, bipolar dorsal kineties, and buccal cirri; frontoterminal, transverse, and caudal cirri are absent. The single species H. pulchra sp. n. has a long, slender body that is brownish in colour, two kinds of cortical granules, one posteriorly located contractile vacuole, one buccal cirrus, usually six frontal cirri, three dorsal kineties, and one midventral row. The main morphogenetic features are: (i) the old oral apparatus is completely replaced by new structures derived from the oral primordium of the proter which originates de novo on the dorsal wall of the buccal cavity, (ii) the posteriormost frontal-midventral-transverse cirral anlage generates a midventral row, (iii) no frontoterminal and transverse cirri are formed, (iv) the anlagen for the marginal rows and the dorsal kineties are formed intrakinetally, and (v) the macronuclear nodules fuse into a mass at the middle stage. Phylogenetic analyses based on SSU rRNA gene sequence data reveal that Heterokeronopsis pulchra is a member of the family Pseudokeronopsidae and is most closely related to Nothoholosticha fasciola.     

Morphology and morphogenesis of a new marine cyrtophorid ciliate, Hartmannula sinica nov. spec. (Protozoa, Ciliophora, Cyrtophorida) from China

The living morphology, infraciliature and morphogenesis of a new marine cyrtophorid ciliate, Hartmannula sinica nov. spec., collected from Qingdao, north China, have been investigated. The new species is characterized by: size in vivo 90–130×40–50 μm, body long elliptical in outline, cilium-free field covered with a conspicuous alveolar layer; 24–31 ventral kineties, the rightmost 6–9 of which extend apically; 20–24 nematodesmal rods; about 15 sparsely distributed contractile vacuoles; a yellowish pigment spot is always present near the anterior tip of the cell. Morphogenetic events exhibit a pattern, which is homologous with that of related cyrtophorids. The main features are as follows: (1) preoral and circumoral kineties of opisthe develop from the oral primordium that forms in mid-body from sections of 3 postoral kineties, while parental preoral and circumoral kineties are retained; (2) parental cytostome and nematodesmal rods are resorbed in middle divisional stages and then replaced by newly formed structures; (3) the heteromerous macronucleus unifies in the late divisional stage.     

Morphological,Ultrastructural, and Molecular Characterization of Euplotidium rosati n. sp. (Ciliophora,Euplotida) from Guam

We combined morphological (i.e. live, stained, scanning, and transmission electron microscopy) with morphometric and molecular analysis to describe a ciliate species collected from shallow reefs in Guam, grown, and maintained in our laboratory. The species was recognized as a member of Euplotidium, and compared with established species of the genus: Euplotidium itoi Ito 1958; Euplotidium psammophilus (Vacelet 1961) Borror 1972; Euplotidium arenarium Magagnini and Nobili 1964; Euplotidium helgae Hartwig 1980; Euplotidium prosaltans Tuffrau 1985, and Euplotidium smalli Lei, Choi and Xu, 2002. To obtain more elements to compare the species, new morphometric data and additional SSU rRNA gene sequences of E. itoi and of E. arenarium are reported. On the basis of this comparison, we established the new species Euplotidium rosati that has a cirral pattern composed of 12 frontoventral and six transverse cirri, and lacks the left marginal cirrus. Euplotidium rosati harbors on its dorsal surface epixenosomes, the peculiar extrusive symbionts described in other Euplotidium species. The whole body of our observations together with the analysis of the data available in the literature leads us to propose a redefinition of the genus. The results may also be useful to clarify the tangled relationship between Euplotidium and Gastrocirrhus.     

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.