Parastrombidinopsis minima n. sp. (Ciliophora: Oligotrichia) from the Coastal Waters of Northeastern Taiwan: Morphology and Small Subunit Ribosomal DNA Sequence

ABSTRACT. Parastrombidinopsis minima n. sp. is investigated, using live observations, protargol preparations, and molecular data. In living cells, the ranges of cell length are 85–95 μm, cell width 60–70 μm, and oral diameter 40–50 μm. In protargol‐impregnated specimens, cell length ranges between 43 and 71 μm, cell width between 23 and 42 μm, and oral diameter between 13 and 24 μm. The numbers of external oral polykinetids are 12–16 and of somatic kineties are 11–13. There are always two ovoid macronuclei (9–16 × 4–9 μm). Based on the analysis of morphologic data, the new species can be placed in the family Strombidinopsidae, but based on the small subunit rRNA gene sequence data, the Parastrombidinopsis species are more closely associated with strobilidiids and tintinnids.     

Ultrastructural and Protargol Studies of Lagenophrys callinectes* (Ciliophora: Peritrichida)

Ultrastructural and protargol studies reveal that the trophont of Lagenophrys callinectes, though highly specialized, generally conforms to the basic peritrich structural pattern. Features described for L. callinectes trophonts which are unique for the genus are the fine structure and arrangement of the lorica and lips, the attachment organelle of the peristomial cytoplasm, its attachment to the loricastome walls, and the arrangement of the aboral kinetosomes of the trophont. Lack of a distinct scopularized region, and of a ventral lorica wall also characterize L. callinectes trophonts. The 4-row terminal peniculus, as revealed by protargol staining, differs from the 6-row terminal peniculus of L. nassa suggesting that the patterns of infundibular structure, as revealed by protargol, should be useful in future taxonomic studies of Lagenophrys species.     

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.     

Morphology and Molecular Phylogeny of Trimyema koreanum n. sp., a Ciliate from the Hypersaline Water of a Solar Saltern

ABSTRACT. A new ciliate, Trimyema koreanum n. sp., isolated from hypersaline water (salinity of 293‰) from a solar saltern in Korea, was investigated using live observation, protargol impregnation, and gene sequencing. Trimyema koreanum is about 30 × 13 μm in vivo, has usually 23 longitudinal ciliary rows forming two distinct ciliary girdles visible both in vivo and in protargol impregnation. A third indistinct ciliary girdle as well as a girdle of mucocysts is distinguishable only in impregnated cells. We suggest T. koreanum as a new species, differing from the most similar species, T. marinum, by the presence of two distinct ciliary girdles (T. marinum usually has six ciliary girdles clearly visible in living cells and three anterior spirals that encircle the cell completely). Although the number of known 18S rRNA sequences in the genus Trimyema was limited, the Trimyema group including T. koreanum forms a strong clade. The phylogenetic position confirms that the isolate belongs to the genus Trimyema and is different from previously sequenced species. Trimyema koreanum is able to consume both prokaryotes and small eukaryotes (specifically, the alga Dunaliella sp.).     

Taxonomic Descriptions of One New Species and Redescription of Four Species in the Family Strombidiidae (Ciliophora,Oligotrichida)1

ABSTRACT Five ciliate species collected from the Woods Hole area were examined by protargol silver impregnation and scanning electron microscopy. These ciliates have been shown to sequester and use chloroplasts obtained from flagellate prey. One new species, Strombidium chlorophilum, is described. Four other species, Strombidium capitatum (Leegaard, 1915) Kahl, 1932, Strombidium conicum (Lohmann, 1908) Wulff, 1919, Strombidium acutum (Leegaard, 1915) Kahl, 1932, and Laboea strobila Lohmann, 1908, are redescribed. Characters used in describing the Strombidiidae include cell size and shape, anterior and ventral polykinetids, macronuclear shape and size, the kinetid “girdle,” the ventral kinety, the trichites, and the paroral kinety. The rationale for using these characters as taxonomic criteria is discussed.     

Two Gonostomatid Ciliates from the Soil of Lombardia,Italy; including Note on the Soil Mapping Project

Two gonostomatid ciliates, Gonostomum paronense n. sp. and G. strenuum, isolated from the soil sample of paddy field, Lombardia, Italy, were investigated using live observation and protargol impregnation. Gonostomum paronense n. sp. is mainly characterized by a tailed body, frontoventral cirri arranged in pairs, and presence of pretransverse and transverse cirri. Morphologically and morphometrically, the new species is similar to Gonostomum namibiense in having a tailed body and frontoventral cirral pairs; however, it differs mainly in the number of frontoventral cirral pairs (seven vs. three). Phylogenetic analyses based on the SSU rDNA sequences show that the new species is more closely related to G. namibiense than to G. strenuum, supporting the morphological classification based on the cirral pattern and the tailed body. However, due to the poor nodal support and absence of gene sequence of the type species Gonostomum, a more robust phylogeny of this group still remains unresolved. The biometric data of the Italian population of Gonostomum strenuum overlap with those from other known populations. Both species were collected from the industrial area of Parona, in the framework of the “Soil Mapping, Lombardia” project in which, for the first time in Italy, soil ciliates were used as bioindicators of soil quality.     

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

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

Patterns of basal body addition in ciliary rows in Tetrahymena

Most naked basal bodies visualized in protargol stains on the surface of Tetrahymena are new basal bodies which have not yet developed cilia. The rarity of short cilia is explained by the rapid development of the ciliary shaft once it begins to grow. The high frequency of naked basal bodies (about 50 percent) in log cultures indicates that the interval between assembly of the basal body and the initiation of the cilium is long, approximately a full cell cycle. Naked basal bodies are more frequent in the mid and posterior parts of the cell and two or more naked basal bodies may be associated with one ciliated basal body in these regions. Daughter cells produced at division are apparently asymmetric with respect to their endowment of new and old organelles.     

Notes on Technic: Pkotargol: Old and New

Stocks of protargol of foreign origin are becoming exhausted, and since such protargol is no longer available, the American-made product must be used for histologic staining. Some of the earlier domestic protargol seems not to have given satisfaction in neurohistology; therefore it seemed desirable to compare the staining qualities of the pre-war material with several lots furnished recently by Winthrop-Stearns, Inc., New York City.     

Two Anaerobic Ciliates (Ciliophora,Armophorea) from China: Morphology and SSU rDNA Sequence,with Report of a New Species,Metopus paravestitus nov. spec

The morphology and phylogeny of two metopid ciliates, collected from anaerobic habitats in China, were investigated using live observation, protargol staining method, and SSU rDNA sequencing. The new species Metopus paravestitus nov. spec. can be distinguished by a combination of the following features: oblong cell with densely arranged ectobiotic prokaryotes perpendicular to cell surface, filiform intracytoplasmic structures packed in the anterior portion of the cell. Our work also demonstrates the wide geographical distribution of Metopus es (Müller, 1776) Lauterborn, 1916. The order Metopida is consistently depicted as a paraphylum in SSU rDNA phylogeny. Metopus paravestitus nov. spec. is closely related to its marine congeners than to freshwater forms. The present study confirms once again the non-monophyly of the genus Metopus and genus Metopidae.     

Notes on Technic: Pkotargol: Old and New

Stocks of protargol of foreign origin are becoming exhausted, and since such protargol is no longer available, the American-made product must be used for histologic staining. Some of the earlier domestic protargol seems not to have given satisfaction in neurohistology; therefore it seemed desirable to compare the staining qualities of the pre-war material with several lots furnished recently by Winthrop-Stearns, Inc., New York City.     

The Morphology of Three Loxophyllum Species (Ciliophora,Pleurostomatida) from Southern China,L. lembum sp. n., L. vesiculosum sp. n. and L. perihoplophorum Buddenbrock, 1920, with Notes on the Molecular Phylogeny of Loxophyllum

Two new pleurostomatid ciliates, Loxophyllum lembum sp. n., L. vesiculosum sp. n., and the poorly known L. perihoplophorum Buddenbrock, 1920, isolated from brackish waters in coastal regions of southern China, are described following observations of live cells and protargol‐impregnated specimens. Loxophyllum lembum sp. n. is distinguished by a combination of characters including two macronuclear nodules, 6–9 contractile vacuoles along the ventral margin, 11–14 right and 6–8 left kineties and the presence of cortical granules. Loxophyllum vesiculosum sp. n. differs from its congeners mainly by the unique distribution of contractile vacuoles, several of which lie along the dorsal margin and one on the ventral margin, and 15–21 right and 6–8 left kineties. Loxophyllum perihoplophorum is characterized by its large cell size (350–450 μm long in vivo), 3–5 contractile vacuoles along the dorsal margin in the posterior region of the body, and 19–23 right and 7–9 left kineties. An improved diagnosis of L. perihoplophorum is provided. The SSU rDNA sequence of L. perihoplophorum is reported for the first time and its molecular phylogeny is analyzed. Maximum likelihood and Bayesian inference analyses of SSU rDNA sequence data recover the monophyly both of the order Pleurostomatida and of the genus Loxophyllum.     

Morphology and morphogenesis of a new marine hypotrichous ciliate (Protozoa,Ciliophora, Pseudoamphisiellidae), including a report on the small subunit rRNA gene sequence

The morphology and morphogenesis of a new marine hypotrichous ciliate Pseudoamphisiella elongata sp. nov. isolated from mussel‐farming waters near Qingdao, China, are described based on living and protargol‐impregnated specimens. Morphologically, the new species can be distinguished from its known congeners by its elongate body shape, narrow oral field, having fewer dorsal kineties and caudal cirri, more marginal cirri, and differentiated pretransverse cirri. The identification as a new species is firmly supported by the sequences of the small subunit ribosomal rRNA (SSU rRNA) gene, compared with other known Pseudoamphisiella species, and the phylogenetic analysis. The morphogenetic characteristics can be summarized as follows: (1) the parental adoral zone of membranelles and undulating membranes are entirely rebuilt by the oral primordium, which develops de novo in the outermost region of the cortex; (2) the oral primordium in the opisthe and the frontoventral–transverse (FVT) anlagen in both dividers are formed independently on the cell surface; (3) an ‘extra’ marginal anlage originates to the right of the right marginal anlage, and develops into two or three ‘extra’ marginal cirri; (4) the FVT anlagen develop in the primary mode, and the last FVT streak contributes two migratory cirri (frontoterminal cirri), which are probably resorbed; (5) the right marginal anlagen in both dividers occur close together, independent of the old structure. © 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 158 , 231–243.     

Morphology and Morphogenesis of Fuscheria terricola n. sp. and Spathidium muscorum (Ciliophora: Kinetofragminophora)1

The morphology and morphogenesis of the kinetofragminophoran soil ciliates, Fuscheria terricola n. sp. and Spathidium muscorum Dragesco & Dragesco-Kerneis, 1979, are described. Stained specimens (protargol) are characterized biometrically. The new species differs from the other species of the genus in its body size, body shape, number of kineties, length of extrusomes, and habitat. Both species have telokinetal stomatogenesis, which commences with a proliferation of kinetosomes at those kineties which bear the brosse. Fuscheria terricola does not have a complex perioral ciliature; indeed, it might be that this species has only monokinetids. Thus only a proliferation of kinetosomes and the separation of the kineties takes place in the prospective division furrow. In contrast, S. muscorum differentiates short dikinetid kinetofragments in the region of the division furrow, which are arranged to form the perioral kinety of the opisthe in the intermediate and late stages of the stomatogenesis. The right part of the perioral kinety develops first. This and other studies show that telokinetal stomatogenesis proceeds very differently depending on the differentiation of the oral ciliature; however, detailed studies on the morphogenesis of kinetofragminophoran ciliates are still too few in number for subtypes to be defined.     

红色伪角毛虫的形态学重描述及与相近种的比较(纤毛门,腹毛目)

利用活体观察、蛋白银染色技术对1种海水腹毛目纤毛虫-红色伪角毛虫Pseudokeronopsis rubra的青岛种群之活体形态和纤毛图式做了研究.其特征如下:虫体呈长带状,尾钝圆,身体柔软,高度可曲;皮层颗粒棕黄色,主要呈玫瑰花形分布,其余散布;表膜下具另1种无色、双面凹形颗粒.口围带小膜数为48～55;左、右缘棘毛各1列,中腹棘毛排成典型的锯齿状并延伸至横棘毛附近;额棘毛无明确分化,呈冠状排布并与中腹棘毛列相连续;口棘毛1根,额前棘毛2根;背触毛4列.大核90～140枚.伸缩泡位于体后左侧1/3处.经比较表明,尽管纤毛图式高度相似,P.rubra仍可以很容易地依据体色、伸缩泡的位置、皮层颗粒的大小、颜色、排布等活体特征与相近种区分.说明活体观察在纤毛虫种类鉴定中具有十分重要的作用.     

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.     

Neuronal and non-neuronal properties of neuroblastoma cells.

Three different mouse neuroblastoma cell lines (S20Y, N2CL10, N115) were examined for acetylcholinesterase activity, ganglioside composition, cell processes, and affinity for protargol silver (i.e., argyrophilia). Assays were made on cloned cells, corresponding tumors which developed after subcutaneous injection of $\text{A}\text{J}$ strain male mice with 2 × 10⁶ cells, and primary and secondary cultures. Acetylcholinesterase activity was present in all cells assayed, with maximal activity noted in cloned cells. Ganglioside patterns of neuroblastoma cells differed from those of neural cells, but remained qualitatively unchanged for a given cell line grown in vivo or in vitro. Some cells were stained with protargol silver in primary cultures, but few cells in cloned or secondary cultures, or those in in vivo tissues, were impregnated with protargol silver. These findings show that while neuroblastoma cell lines maintain some neuronal characteristics (i.e., high acetylcholinesterase levels, cell processes), they do not express other accepted neuronal properties (i.e., ganglioside patterns, argyrophilia), and suggest that direct analogies between normal neurons and “differentiated” neuroblastoma cells should be made with caution.     

Morphogenesis and Molecular Phylogeny of a New Freshwater Ciliate,Notohymena apoaustralis n. sp. (Ciliophora,Oxytrichidae)

The live morphology, infraciliature, and morphogenesis of a new oxytrichid ciliate, Notohymena apoaustralis n. sp. collected from a freshwater pond in Qingdao (Tsingtao), China, were studied in vivo and after protargol impregnation. Notohymena apoaustralis n. sp. is characterized as follows: undulating membranes in Notohymena‐pattern; cortical granules yellow‐green, grouped around the marginal cirri and dorsal bristles, and in short irregular rows elsewhere in the cell; single contractile vacuole positioned at anterior 1/3 of the body length; two macronuclear nodules and one micronucleus; about 39 adoral membranelles; 18 frontoventral transverse cirri in typical Oxytricha‐pattern; one right and one left marginal row, almost confluent posteriorly; dorsal ciliature in typical Oxytricha‐pattern; 8–10 caudal cirri arranged in three rows, one each at the posterior end of dorsal kineties 1, 2, and 4, indistinguishable from marginal cirri in life. The morphogenetic process in N. apoaustralis n. sp. is consistent with that of the type species, Notohymena rubescens Blatterer and Foissner, 1988. Phylogenetic analyses based on small subunit rDNA sequence data suggest a sister relationship between N. apoaustralis n. sp. and Paraurostyla weissei, which cluster in a clade with Rubrioxytricha ferruginea.     

A Somatic Kinetid Study of the Pycnotrichid Ciliate Vestibulongum corlissi N. G., N. Sp. (Class: Litostomatea), Symbiont in the Intestines of the Surgeonfish,Acanthurus xanthopterus1

ABSTRACT The ciliate Vestibulongum corlissi n. g., n. sp. was collected from the intestines of surgeonfish, Acanthurus xanthopterus, in the summer of 1986. In has been examined in the light microscope after protargol staining and by scanning and transmission electron microscopy. Its form is distinct from that of other pycnotrichid ciliates at the generic level. Somatic kinetids were examined; these demonstrate that its cytostome is posterior and that the kinetid structures and the presence of a second transverse microtubular ribbon confirm its placement in the class Litostomatea.