Reevaluation of cortical developmental patterns in Euplotes (s. l.), including a morphogenetic redescription of E. charon (Protozoa, Ciliophora, Euplotida)

We documented the pattern of cell development in Euplotes charon. The ontogenesis of this species was similar to many of its congeners, except for the formation of the caudal cirri. In E. Charon, a caudal cirrus is formed posterior to each of the rightmost two or three dorsal kinety anlage in the proter, and the second rightmost dorsal kinety in the opisthe. In addition, two caudal cirri are formed posterior to the rightmost dorsal kinety in the opisthe. This pattern of development represents a completely new type. Based on our evaluation, and in comparison with previous studies, we also conclude that the pattern of cell development is variable among species in the Euplotes genera. The variation is particularly evident during the formation of frontoventral and caudal cirri. Based on the segmentation pattern of frontal-midventral transverse cirral anlagen, cirri reduction, and migration of frontoventral cirri, we identified five types: the affinis-type, the eurystomus-type, the charon-type, the raikovi-type and orientalis-type. Euplotes (s. l.) can also be divided into three types based on the formation of caudal cirri: focardii-type, vannus-type and charon-type. Indeed, we conclude that the number (one or two) of marginal cirri should be given as much consideration as the genetic separation. Given this, we reassessed the validity of using genetic separation to classify the group. Generally, the morphogenetic data disagreed with the molecular data (SSrRNA gene sequences). Given these discrepancies, it is too early to draw conclusions on the systematic arrangement of this species-rich taxon.     

Taxonomic description of a new marine ciliate,Euplotes qingdaoensis n. sp. (Ciliophora: Euplotida)

The characteristics and infraciliature of a new euplotid ciliate,Euplotes qingdaoensis n.sp.,discovered from the coastal waters of Jiaozhou Bay,northern China,were studied using live observation and silver impregnation.The new marine species is characterized by a slightly fl attened body(90–110 μm×70–80 μm,in vivo),an adoral zone of 26–29 membranelles,10 strong frontoventral and fi ve transverse cirri,two caudal cirri,a single left marginal cirrus,seven dorsal kineties,a middle kinety with about 15 pairs of basal bodies,a silverline system of the double-eurystomus type,and a C-shaped macronucleus.     

Population dynamics of marine ciliate Euplotes vannus (Protozoa, Ciliophora) in different artificial seawaters

To study population dynamics of marine ciliates in different artificial seawaters (ASW), the population growth dynamics of a common marine ciliate Euplotes vannus were investigated using beef extract media and rice media for five types of ASW and natural seawater (NSW). The results show that: (1) the population growth rate was in the order of NSW>Flack ASW>Nakamula ASW>Schmadz ASW>Oshima ASW>Subow ASW and was considerably higher in rice media than in beef extract media (apart from Subow ASW); (2) the maximum density of E. vannus in stationary phase in each treatment was ranked as Flack ASW>Nakamula ASW>Schmadz ASW>NSW>Oshima ASW>Subow ASW, and was again higher in rice media than in beef extract media (except for Subow ASW); (3) the exponential and stationary phases were longer in rice media than in beef extract media; (4) strains of E. vannus that had been domesticated for >1 year in ASW grew significantly slower, with lower maximum density and longer stationary phase than those isolated and maintained in NSW. It was demonstrated that: (1) E. vannus may grow well in Flack, Nakamula and Schmads ASW compared with NSW (mainly in terms of growth rate); and (2) Oshima ASW is the preferred choice for stock cultures of E. vannus, but the ASWs Flack, Nakamula and Schmadz are preferred for mass culture. These findings suggest that these three ASWs are effective for the cultivation of marine protozoa for experimental studies on ecology, toxicology and molecular biology.     

Phylogenetic relationships within the genus Aspidisca (Protozoa, Ciliophora, Euplotida) revealed by ITS1-5.8S-ITS2 region sequences

The internal transcribed spacer regions (ITS1 and ITS2) and 5.8S rRNA genes were sequenced in six populations of four Aspidisca species, namely A. leptaspis, A. orthopogon, A. magna and A. aculeata. Phylogenetic trees were constructed by means of Bayesian inference (BI), Maximum Parsimony (MP), Neighbor-Joining (NJ), and Maximum Likelihood (ML) to assess the inter- and intra-species relationships within the genus Aspidisca. All trees show similar topologies with stable supports and indicate that: (1) four well known groups, i.e., Oligotrichia, Stichotrichia, Choreotrichia and Hypotrichia, are distinctly outlined within the class Spirotrichea, and all are monophyletic other than Hypotrichia; (2) members of Aspidisca can be distinguished well, based on the ITS1-5.8S-ITS2 region sequences, and A. leptaspis and A. magna shared a closer relationship than other species; (3) Aspidisca and Euplotes branch early in the subclass Hypotrichia. To compare the phylogenetic relationships based on different genes, SSU rRNA trees were also constructed with nearly the same species inclusion, which revealed different topologies of inter-species, inter-genera and inter-subclasses.     

Phylogeny of subclass Scuticociliatia (Protozoa, Ciliophora) using combined data inferred from genetic, morphological, and morphogenetic evidence

Gene sequence-based genealogies of scuticociliates are different from those produced by morphological analyses. For this reason, 11 representative scuticociliates and two ambiguously related genera were chosen to test the ability of combined phylogenetic analyses using both gene sequences and morphological/morphogenetic characteristics. Analyses of both the SSrRNA gene sequences and the combined datasets revealed a consistent branching pattern. While the terminal branches and the order level relationships were generally well resolved, the family level relationships remain unresolved. However, two other trees based on ITS1-5.8S-ITS2 region sequences and morphological/morphogenetic characters showed limited information, due to a lack of informative sites in these two datasets. Our data suggest, however, that the combined analysis of morphological/morphogenetic characters and gene sequences did produce some changes to the phylogenetic estimates of this group.     

A new marine ciliate,Amphileptus litonotiformis nov. SP. (Protozoa Ciliophora)

The morphology and infraciliature of a marine ciliateAnphileptus litonotiformis nov. sp. inhabiting a shrimp farming pond in Sheyang (Jiangsu Province) are described. According to the infraciliature, habitat, and other morphological characters, it can not be congeneric with any other species of the genus. The special features ofA. litonotiformis are theLitonotus-like body shape and the arrangement of its extrusomes regularly located along the margin of the cytostome. The most important characters of some relatedAnphileptus-species from marine water are biometrically tabled and discussed.     

Report and Phylogeny of a Novel Euplotes Species from China: E. mazeii n. sp., and Review of E. balteatus(Alveolata,Ciliophora)

The freshwater species, Euplotes mazeii n. sp., and the marine species, E. balteatus(Dujardin, 1841) Diesing, 1850 were collected from China and investigated based on their living morphology, ciliary pattern, and small subunit ribosomal RNA(SSU rRNA)gene sequence data. Euplotes mazeii n. sp. was characterized by its small cell size((40 – 55) μm ×(25 – 35) μm), nine frontoventral cirri,one marginal and two caudal cirri, seven dorsal kineties, and a double-eurystomus type of dorsal silverline syst...     

A general overview of the typical 18 frontal-ventral-transverse cirri oxytrichidae s. l. genera (Ciliophora,Hypotrichia)

Oxytrichidaes. l.ciliates usually have 18 frontal-ventral-transverse cirri which are clustered to six distinct groups usu-ally originating from six longitudinal primordia segregating 1, 3, 3, 3, 4, 4 c...     

Tolerance of ciliated protozoan Paramecium bursaria （Protozoa,Ciliophora） to ammonia and nitrites

The tolerance to ammonia and nitrites in freshwater ciliate Paramecium bursaria was measured in a conventional open system. The ciliate was exposed to different concentrations of ammonia and nitrites for 2h and 12h in order to determine the lethal concentrations. Linear regression analysis revealed that the 2h-LC50 value for ammonia was 95.94 mg/L and for nitrite 27.35 mg/L using probit scale method (with 95% confidence intervals). There was a linear correlation between the mortality probit scale and logarithmic concentration of ammonia which fit by a regression equation y=7.32x-9.51 (R^2=0.98; y, mortality probit scale; x, logarithmic concentration of ammonia), by which 2 h-LC50 value for ammonia was found to be 95.50 mg/L. A linear correlation between mortality probit scales and logarithmic concentration of nitrite is also followed the regression equation y=2.86x 0.89 (R^2=0.95; y, mortality probit scale; x, logarithmic concentration of nitrite). The regression analysis of toxicity curves showed that the linear correlation between exposed time of ammonia-N LC50 valueand ammonia-N LC50 value followed the regression equation y=2 862.85e-0.08x (R^2=0.95; y, duration of exposure to LC50 value; x, LC50 value), and that between exposed time of nitrite-N LC50 value and nitrite-N LC50 value followed the regression equation y = 127.15e-0.13x (R^2=0.91; y, exposed time of LC50 value; x, LC50 value). The results demonstrate that the tolerance to ammonia in P. bursaria is considerably higher than that of the larvae or juveniles of some metozoa, e.g. cultured prawns and oysters. In addition, ciliates, as bacterial predators, are likely to play a positive role in maintaining and improving water quality in aquatic environments with high-level ammonium, such as sewage treatment systems.     

Tolerance of ciliated protozoan Paramecium bursaria (Protozoa, Ciliophora) to ammonia and nitrites

1 INTRODUCTION Ammonia and nitrites are common pollutants in waters used for intensive aquiculture, and are harmful to cultured animals and their larvae when present in sufficiently high concentrations (Zhou, 1991). Ammonia in water is usually derived from animal excretion and catabolism of organic materi- als. It occurs in un-ionized and ionized forms. Un-ionized ammonia can pass through cell mem- brane and is harmful to the cells due to its fat-soluble feature (Chen, 2000; Chin, 2001).…     

Molecular Phylogenetic Lineage of Plagiopogon and Askenasia (Protozoa,Ciliophora) Revealed by Their Gene Sequences

Prostomates and haptorians are two basal groups of ciliates with limited morphological characteristics available for taxonomy. Morphologically, the structures used to identify prostomates and haptorians are similar or even identical, which generate heavy taxonomic and phylogenetic confusion. In present work, phylogenetic positions lineage of two rare genera, Plagiopogon and Askenasia, were investigated. Three genes including small subunit ribosomal RNA gene(hereafter SSU r DNA), internal transcribed spacer region(ITS region), and large subunit ribosomal RNA gene(LSU r DNA) were analyzed, 10 new sequences five species each. Our findings included 1) class Prostomatea and order Haptorida are multiphyletic; 2) it may not be appropriate to place order Cyclotrichiida in subclass Haptoria, and the systematic lineage of order Cyclotrichiida needs to be verified further; 3) genus Plagiopogon branches consistently within a clade covering most prostomes and is basal of clade Colepidae, implying its close lineage to Prostomatea; and 4) Askenasia is phylogenetically distant from the subclass Haptoria but close to classes Prostomatea, Plagiopylea and Oligohymenophorea. We supposed that the toxicyst of Askenasia may be close to taxa of prostomes instead of haptorians, and the dorsal brush is a more typical morphological characteristics of haptorians than toxicysts.     

Taxonomy and Phylogeny of a New Marine Planktonic Ciliate,Strombidium pseudorapulum sp. n.(Protozoa,Ciliophora, Oligotrichia)

As one of the main groups of marine planktonic protozoa, oligotrich ciliates have shown a high biodiversity. Recently, a new aloricate oligotrich ciliate, Strombidium pseudorapulum sp. n. was isolated from coastal waters of southeast China. The living morphology and infraciliature of this species were studied using live cell observation and protargol impregnation. Strombidium pseudorapulum sp. n. differs from its congeners by having a radish-like body with a bumped anterior end and an elongated tail-like posterior end, possessing a red ‘eyespot' in apical protrusion of cell, adoral zone of membranelles composed of 12–16 collar and 9–12 buccal membranelles, one spindle-shaped macronucleus, girdle kinety in anterior 2/5 of cell with 45–60 densely spaced dikinetids, and ventral kinety occupying about the posterior 1/3 of the cell with 16–19 dikinetids. Strombidium pseudorapulum sp. n. clusters together with those species having ‘eyespot' in apical protrusion in the phylogenetic tree based on the small subunit rRNA gene sequence. It is a sister species of S. cuneiforme with 31 unmatched nucleotides between their SSU rRNA gene sequences.     

Molecular phylogeny of oligotrich genera Omegastrombidium and Novistrombidium (Protozoa, Ciliophora) for the systematical relationships within Family Strombidiidae

The phylogeny of the oligotrich ciliates is currently a hot debate despite the availability of both morphological and molecular data. In the present paper, further small subunit rRNA (SS rRNA) genes were analyzed from the Genera Omegastrombidium and Novistrombidium, as well as from Strombidium, and combined with three new SS rRNA sequences from Strombidium basimorphum, S. sulcatum population QD-1, and Novistrombidium testaceum population GD. The phylogenetic positions of these organisms were inferred using Bayesian inference, Maximum Likelihood, and Maximum Parsimony methods. The main results are: (1) the SS rRNA gene sequence analyses match the recent findings about the molecular evolution of oligotrichs, indicating that the family Strombidiidae is paraphyletic; (2) the Genus Omegastrombidium is separated from the Genus Strombidium, as shown in recent cladistic analyses; (3) morphospecies in Genus Novistrombidium, based on similarity of somatic ciliature, are separated from each other in all topological trees, indicating that this genus could be a paraphyletic group; (4) the molecular data indicate a possibility of paraphyly for the genus Strombidium; and (5) the similarities of the SS rRNA gene of specimens identified as S. sulcatum and S. inclinatum are 99.8%–100%. However, present knowledge on the oligotrichs sensu stricto is still insufficient and further studies based on both molecular and other technologies are required.     

Morphology and SSU rD NA Sequence Analysis of Two Hypotrichous Ciliates(Protozoa,Ciliophora, Hypotrichia) Including the New Species Metaurostylopsis parastruederkypkeae n.sp.

The morphology and phylogeny of two hypotrichous ciliates, Metaurostylopsis parastruederkypkeae n. sp. and Neourostylopsis flavicana(Wang et al., 2011) Chen et al., 2013 were investigated based on morphology, infraciliature and the small subunit(SSU) ribosomal RNA gene(r RNA) sequence. The new species, M. parastruederkypkeae n. sp. was identified according to its characteristics: body shape ellipsoidal, size about(165–200) ×(45–60) μm in vivo, cell color reddish; two types of cortical granules including wheat grain-like and yellow-greenish larger ones along the marginal cirri rows and dorsal kineties and dot-like and reddish smaller ones, grouped around marginal cirri on ventral side and arranged in short lines on dorsal side; 26–41 adoral membranelles; three frontal and one parabuccal, five to seven frontoterminal, one buccal, and three to six transverse cirri; seven to thirteen midventral pairs; five to nine unpaired ventral cirri, five to seven left and three to five right marginal rows; and three complete dorsal kineties. Phylogenetic analysis based on SSU r DNA sequences showed that both Metaurostylopsis and Neourostylopsis are monophyletic. As the internal relationship between and within both genera are not clear, further studies on the species in these two genera are necessary. The key characteristics of all known twelve Metaurostylopsis-Apourostylopsis-Neourostylopsis species complex were updated.     

Morphology and SSU rDNA sequence analysis of two hypotrichous ciliates (Protozoa,Ciliophora, Hypotrichia) including the new species <Emphasis Type="Italic">Metaurostylopsis parastruederkypkeae</Emphasis> n. sp.

The morphology and phylogeny of two hypotrichous ciliates, Metaurostylopsis parastruederkypkeae n. sp. and Neourostylopsis flavicana (Wang et al., 2011) Chen et al., 2013 were investigated based on morphology, infraciliature and the small subunit (SSU) ribosomal RNA gene (rRNA) sequence. The new species, M. parastruederkypkeae n. sp. was identified according to its characteristics: body shape ellipsoidal, size about (165–200) × (45–60) μm in vivo, cell color reddish; two types of cortical granules including wheat grain-like and yellow-greenish larger ones along the marginal cirri rows and dorsal kineties and dot-like and reddish smaller ones, grouped around marginal cirri on ventral side and arranged in short lines on dorsal side; 26–41 adoral membranelles; three frontal and one parabuccal, five to seven frontoterminal, one buccal, and three to six transverse cirri; seven to thirteen midventral pairs; five to nine unpaired ventral cirri, five to seven left and three to five right marginal rows; and three complete dorsal kineties. Phylogenetic analysis based on SSU rDNA sequences showed that both Metaurostylopsis and Neourostylopsis are monophyletic. As the internal relationship between and within both genera are not clear, further studies on the species in these two genera are necessary. The key characteristics of all known twelve Metaurostylopsis-Apourostylopsis-Neourostylopsis species complex were updated.     

Taxonomy,Morphology and Phylogeny of a New Oligotrich Ciliate-Omegastrombidium hongkongense n. sp.(Protozoa: Ciliophora) from Clear Water Bay,Hong Kong

One new marine oligotrich ciliate, Omegastrombidium hongkongense n. sp., was isolated from a bloom of Noctiluca scuntillans near Port Shelter, Hong Kong. The morphology and infraciliature of this new species were studied on both living and protargol-stained specimens. Its phylogenetic position was discussed based on the sequence of the small subunit r RNA gene. O. hongkongense is different from its congeners with special characters. The cells are usually heart-shaped, and the cell size usually is(20–35) ×(20–30) μm in vivo. Its deep buccal cavity extends obliquely to about 1/2 of cell length. It shows prominent apical protrusion. The adoral zone of membranelles is divided into 17–19 collar membranelles and four buccal membranelles. It has one ball-like macronucleus. The girdle kinety forms a closed loop which obliquely surrounds the body. The ventral kinety and thigmotactic membranelles are not observed. The SSU r RNA sequence of O. hongkongense was close to those of Strombidium paracalkinsi and Varistrombidium kelum with approximately 99% similarity. In the phylogenetic trees, O. hongkongense can be grouped with O. elegans and V. kielum species with very low support(16% ML).