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 and morphogenesis of a new oxytrichid ciliate,Notohymena limus n. sp. (Ciliophora,Oxytrichidae) from Delhi,India

The morphology and morphogenesis of a new oxytrichid ciliate, Notohymena limus n. sp. were studied in vivo and after protargol impregnation. The new ciliate was isolated from the sewage sludge at Delhi Jal Board Sewage Treatment Plant located at Rithala, Delhi, India, using the non-flooded Petri dish method. N. limus n. sp. is characterized as follows: flexible dorsoventrally flattened ellipsoidal body; Notohymena-pattern undulating membranes; adoral zone of membranelles (AZM) occupied about 39% of the body length, and consists of around 26 membranelles; large and deep buccal cavity; colorless subpellicular granules present in groups and arranged around the bases of dorsal bristles; 4 macronuclear nodules; 2 micronuclei; 18 fronto-ventral-transverse (FVT) cirri in typical Oxytricha-pattern; 6 dorsal rows of bristles; 3 caudal cirri; about 16 right and 15 left marginal cirri; N. limus n. sp. is a new species on the basis of the combination of morphological, morphometric and morphogenetic characteristic features.     

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 brackish water ciliate Bakuella subtropica sp. n. (Ciliophora,Hypotricha) from southern China

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

Morphology and morphogenesis of Apoholosticha sinica n. g., n. sp. (Ciliophora,Hypotrichia), with consideration of its systematic position among urostylids

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

Molecular Phylogeny and Taxonomy of a New Freshwater Hymenostomatid from Northeastern China,with the Establishment of a New Genus Anteglaucoma gen. n. (Protista,Ciliophora, Oligohymenophorea)

The morphology, infraciliature and SSU rDNA sequence of a new freshwater hymenostomatid ciliate, Anteglaucoma harbinensis gen. nov., spec. nov., collected from a farmland pond in Harbin, China, were investigated. The new genus Anteglaucoma is characterized as follows: small to medium‐sized Glaucomidae with oral apparatus in anterior one‐third of cell; paroral membrane composed of almost longitudinally arranged dikinetids; three adoral membranelles nearly equal in length and arranged almost longitudinally in parallel; silverline pattern tetrahymenid. The improved diagnosis of family Glaucomidae Corliss 1971 is provided based on the previous and present work. The type species Anteglaucoma harbinensis spec. nov. is defined by having 32–35 somatic kineties; four or five postoral kineties; membranelle 1 and membranelle 2 having five or six kinetosomal rows, membranelle 3 having three kinetosomal rows; single macronuclear nodule; contractile vacuole on average 15% from posterior body end; locomotion characterized by crawling with a rather hectic jerking motion; freshwater habitat. Phylogenetic analyses show that Anteglaucoma clusters in the family Glaucomidae and groups with the genera Glaucoma. The molecular and morphological data indicate that Glaucomidae is related to the family Bromeliophryidae in the phylogenetic trees.     

Morphology,morphogenesis, and molecular phylogeny of a novel saline soil ciliate,Heterourosomoida sinica n. sp. (Ciliophora,Hypotrichia)

The living morphology, infraciliature, morphogenesis and phylogenetic position of a new hypotrichous ciliate, Heterourosomoida sinica n. sp., discovered in saline soil in Northwest of China, were investigated. Heterourosomoida sinica n. sp. is characterized as follows: body size about 70–95 × 20–30 μm in vivo, ellipsoid or elongate; cortical granules dark green, 0.5 μm across, arranged around dorsal cilia and in short irregular rows on ventral side; consistently 18 frontoventral-transverse cirri; left and right marginal rows composed of 17–24 and 13–21 cirri, respectively; two macronuclear nodules and one or two micronuclei. The detailed morphogenesis of a species of Heterourosomoida is reported for the first time. The ontogenesis on dorsal side is basically similar to that in Urosomoida-species and the process on ventral side is characterized by: 18 frontoventral-transverse cirri develop from six anlagen with the anlagen V and VI formed in the primary mode. Phylogenetic analyses based on SSU rDNA sequence data reveal that Heterourosomoida sinica n. sp. clusters with Kleinstyla dorsicirrata and forms a clade with the type species H. lanceolata.     

Morphology,morphogenesis, and molecular phylogeny of a new freshwater ciliate,Gonostomum jangbogoensis n. sp. (Ciliophora,Hypotricha), from Victoria Land,Antarctica

In a study on ciliate diversity, we discovered the new hypotrich species, Gonostomum jangbogoensis n. sp., in freshwater from Terra Nova Bay, Victoria Land, southeast Antarctica. We describe its morphology and morphogenesis using standard methods, and the SSU rRNA gene phylogeny is provided as well. Morphology of Gonostomum jangbogoensis n. sp. is characterized as follows: slender to elongated body shape; grayish under low magnification; cortical granules present; 32–41 adoral membranelles; 3 enlarged frontal cirri; 1 buccal cirrus; 2 frontoterminal cirri; 3 or 4 frontoventral cirral pairs, 2 pretransverse cirri, 6–7 transverse cirri; 13–19 left and 18–26 right marginal cirri; 17–23 paroral kinetids; 3 dorsal kineties; 3 caudal cirri; 2 macronuclear nodules with 1–3 micronuclei. The morphogenesis of the new species confirms that it has at least seven frontal-ventral-transverse cirral anlagen, which is also reported in Gonostomum sp. 1 sensu Shin from Korea. Even though these two populations occur very far from each other, the morphometric data prove that this character state, the seven cirral anlagen, is a stable feature across these populations and might be an apomorphy. The phylogenetic analyses show that the genus Gonostomum is non-monophyletic and that the new species is a sister to G. bromelicola.     

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.     

Morphology,morphogenesis and molecular phylogeny of Lamtostyla granulifera sinensis subsp. nov. (Ciliophora,Hypotrichia) from a wetland in China

The morphology, morphogenesis and molecular phylogeny of a hypotrichous ciliate, Lamtostyla granulifera sinensis subsp. nov., isolated from northern China, were investigated. This population appeared highly similar in morphology to L. granulifera Foissner, 1997. However, on detailed investigation some non-overlapping features were identified, i.e., the body shape and the arrangement of the cortical granules. These differences suggested the separation at subspecies level. Furthermore, the morphogenesis of the new subspecies is described, which is characterized by: (1) the posterior part of the parental adoral zone of membranelles is renewed; (2) the amphisiellid median cirral row is formed from two anlagen; and (3) the frontoventral-transverse cirral anlagen II to VI generate one transverse cirrus each. Phylogenetic analyses based on SSU rDNA sequence data show that Lamtostyla species are scattered in different clades. The monophyly of the genus Lamtostyla is also rejected by the AU test in this study.     

Morphology,ontogenesis and molecular phylogeny of a new saline soil ciliate,Uroleptoides salina nov. spec. (Ciliophora,Hypotrichia)

The morphology, morphogenesis and molecular phylogeny of a new saline soil hypotrich ciliate, Uroleptoides salina nov. spec., discovered from China, was investigated. The new species is characterized as follows: body 150–215 × 40–50 μm in vivo, slender and highly flexible; usually four ellipsoidal macronuclear nodules; contractile vacuole absent; cortical granules absent; endosymbiotic algae present; amphisiellid median cirral row consists of 14–25 cirri and terminates about 47% down length of body; usually three buccal cirri and 3–13 cirri left of anterior portion of amphisiellid median cirral row; 3–5 transverse cirri. Morphogenesis during binary fission is characterized by: (1) the parental adoral zone of membranelles is retained completely, parental paroral contributes to the formation of the undulating membranes anlage for the proter; (2) the oral primordium of the opisthe is formed apokinetally; and (3) the amphisiellid median cirral row is formed from two anlagen. Phylogenetic analyses based on SSU rDNA sequence data show that Uroleptoides salina nov. spec. has a close relationship with its morphologically similar species, U. longiseries, U. magnigranulosus, Orthamphisiella breviseries, and Parabistichella variabilis.     

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.     

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.     

Morphology,morphogenesis and molecular phylogeny of a novel saline soil ciliate,Urosoma quadrinucleatum n. sp. (Ciliophora,Hypotrichia)

The morphology and morphogenesis of a new saline soil hypotrichous ciliate, Urosoma quadrinucleatum n. sp., collected from northwestern China, were studied based on live observations and protargol stained specimens. The new species is characterized as follows: size in vivo 90–130 × 20–30 μm; body outline elongate-elliptical with both ends broadly rounded; four macronuclear nodules; cortical granules present; paroral in front of endoral; usually 16 frontal-ventral-transverse cirri. Urosoma quadrinucleatum n. sp. has almost the same morphogenetic characteristics as its congeners U. gigantea and U. salmastra, and differs from other three congeners whose morphogenesis is known in the formation of the frontal-ventral-transverse cirral anlagen as well as the development of marginal and dorsal kineties anlagen. The sequence differences among U. quadrinucleatum n. sp. and other Urosoma species further support the validity of the present organism as novel species. Further, U. quadrinucleatum n. sp. clusters with U. salmastra in the phylogenetic analyses based on small subunit ribosomal RNA gene (SSU rDNA) sequence data.     

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.     

Morphology and phylogeny of Vannella croatica n. sp. (Amoebozoa,Discosea, Vannellida)

We isolated and described a new species of freshwater vannellid amoeba from Krka natural reserve in Croatia – Vannella croatica n. sp. This species has certain morphological differences from all known vannellids and differs at the level of SSU sequence. It resembles in size and morphology Vannella lata; to facilitate direct comparison we publish images of V. lata CCAP 1589/12 strain (type strain, which is now lost) taken in 1999. Vannela croatica feeds on bacteria and can be easily grown in large amount in relatively pure culture and thus is suitable for molecular and biochemical studies requiring large amounts of material.     

Anteholosticha foissneri n. sp., a marine hypotrich ciliate (Ciliophora: Spirotrichea) from Vietnam: Morphology,morphogenesis, and molecular phylogeny

In a study of marine ciliate diversity, we collected an Anteholosticha monilata-like population from Vietnam. To identify this population, we analyzed its morphology, some morphogenetic stages, and molecular phylogeny. Based on these data, we conclude that the Vietnamese population is new to science. Anteholosticha foissneri n. sp. resembles Anteholosticha monilata-like species considering (1) the number and arrangement of macronuclear nodules and micronuclei; (2) the presence of cortical granules; and (3) the saline habitat. However, the new species can be easily distinguished from these species by the arrangement, color, and shape of the cortical granules. The divisional morphogenesis commences with the de novo proliferation of basal bodies as a single longitudinal patch left of the posteriormost midventral cirral pair. This character state has not been reported before in Anteholosticha (based on check of the available data) and probably reflects a distinct clade within the nuclear small subunit ribosomal RNA gene tree.     

Morphology and small subunit rRNA gene sequence of Uronemita parabinucleata n. sp. (Ciliophora,Uronematidae), with an improved generic diagnosis

The morphology and infraciliature of a new species, Uronemita parabinucleata n. sp., isolated from intertidal sediments in a coastal region in northern China, were investigated using live observation and silver impregnation methods. The new species is characterized by an in vivo body size of about 20–50 × 10–25 μm, 22 or 23 somatic kineties, two macronuclear nodules, and one caudal cilium. Its small subunit ribosomal RNA gene (SSU rDNA) was sequenced and compared with those of other Uronemita species to reveal nucleotide differences. Phylogenetic analyses indicated that Uronemita is monophyletic and that the new species clusters with its congener Uronemita filificum, with full support provided by both Bayesian inference and maximum likelihood algorithms. Based on previous studies and the present study, an improved diagnosis of the genus Uronemita is supplied, which has been absent since the establishment of this genus. A key to the Uronemita species is also provided.