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To clarify the confusion surrounding species identification and state intraspecific variations, this research studied the morphological taxonomy of five species, Kentrophyllum cf. setigerum Petz et al., 1995; Zosterodasys shumerica Aliev, 1990; Frontonia tchibisovae Burkovsky, 1970; Neourostylopsis orientalis Chen et al., 2013; and Apoholosticha cf. sinica Fan et al., 2014, which were collected from Chongming Island's marsh wetlands. Kentrophyllum cf. setigerum possesses less somatic kineties than K. setigerum (19–26 right and 18–25 left kineties vs 35–40 right and 30 left kineties). Whether Z. shumerica is identical to Z. transversa Foissner et al., 1994, which was presented in a previous report, was negated. The improved diagnosis for the species is provided. Frontonia tchibisovae was confirmed to have three or four micronuclei, and the number of kinetosome rows in its peniculus 3 varied between China and Russia populations. The new population of N. orientalis bears cortical granules, which differs from the original population that has an arrangement of rows across the dorsal bristles. For A. cf. sinica, there are colorless cortical granules on the ventral side of the new population, whereas in the previously reported populations bright orange, spherical granules were clustered around dorsal cilia. Additionally, we sequenced the small-subunit (SSU) rRNA genes of K. cf. setigerum, N. orientalis, and A. cf. Sinica, and compared the new and reported populations. For the first time, we also provide a species list, including 33 ciliates collected from the wetlands based on living observation and silver staining. [ABSTRACT FROM AUTHOR]

Abstract: The morphologies of two novel ciliates, Arcanisutura chongmingensis n. gen., n. sp. and Naxella paralucida n. sp., collected from Shanghai, China, have been investigated using live observation and silver staining methods. Arcanisutura n. gen. can be easily distinguished from related genera by its inconspicuous, oblique anterior suture. Arcanisutura chongmingensis n. sp. is mainly recognized by its elongated body with a tail‐like posterior end, 25–33 somatic kineties, and 4–11 excretory pores. Naxella paralucida n. sp. can be distinguished from its congeners based on its two short nassulid organelles, fusiform trichocysts, 37–49 somatic kineties, and 16 nematodesmal rods. The small‐subunit (SSU) rRNA gene sequences of these two species are presented, revealing the phylogenetic positions of Arcanisutura and Naxella. Phylogenetic analyses show that Arcanisutura forms a sister clade to other synhymeniid genera, namely, Chilodontopsis, Orthodonella, and Zosterodasys; Naxella is most closely related to Nassula spp. and is located within the monophyletic clade of the family Nassulidae. [ABSTRACT FROM AUTHOR]

Ciliates are able to form resting cysts as a survival strategy in response to stressful environmental factors. Studies on the characteristics of cellular structure during encystment may provide useful information for further understanding of the regulatory mechanism of cellular patterns and supply new clues regarding the phylogeny of ciliates. Scanning and transmission electron microscopies were used to observe the ultrastructure of cells during encystment of the soil ciliateAustralocirruscf.australis. The dedifferentiation of ciliature was revealed for the first time. Ciliary shafts first shortened, and the remaining ciliature, including basal bodies and the fibrillar cirral basket, retracted into the cytoplasm and was surrounded by the autophagic vacuoles and then gradually digested. A large number of autophagic vacuoles were observed in mature resting cysts. Autophagy might not only be necessary for the differentiation of cellular structures during encystment but might also be important to sustain the basic life activities in the resting stage.Australocirruscf.australisformed a kinetosome-resorbing cyst and contained four layers in the cyst wall: the ectocyst, mesocyst, endocyst and granular layer. The ciliature resorbing state and the number of layers in the cyst wall were consistent with those found in other oxytrichous ciliates. However, the phenomenon wherein the two macronuclear nodules are not fused during encystment is not commonly observed among oxytrichids. Additionally, the octahedral granules in the mesocyst of this species exhibit different morphology from the congeners. [ABSTRACT FROM PUBLISHER]

Previously, a series of Simocephalus taxa (Cladocera: Daphniidae) from China were described. Most were proposed to be junior synonyms in the last revision of the genus. Using original material from China and data from GenBank, we investigate the biodiversity and phylogeny of Simocephalus using sequences of the cytochrome c oxidase subunit I (COI) and the nuclear 18S genes. In both cases, neighbor-joining, maximum likelihood and Bayesian inference analyses led to highly congruent tree topologies. The grouping of the deeper clades agrees with the inter-generic classification of Orlova-Bienkowskaja (2001). Only the populations of S. serrulatus from Eurasia and North America seem to be closely related, and there are no other shared species between the two continents. Our study unambiguously confirms the existence of many lineages from the subgenera of Simocephalus (Echinocaudus) and Simocephalus s.str. in China, but their morphology needs to be reexamined by taking a wider range of characters (e.g., of female thoracic limbs and adult males) into consideration. [ABSTRACT FROM AUTHOR]

Three species of Microthoracids, Lopezoterenia paratorpens n. sp., Trochiliopsis australis Foissner et al., 1988 and Discotricha papillifera Tuffrau, 1954, collected from Chinese coastal waters, were investigated using live observation and protargol staining methods. Lopezoterenia paratorpens n. sp. was characterized by its squarely shaped cortical papillae and by dorsal kineties which contained loosely distributed basal bodies. Trochiliopsis australis was revealed to have two oral membranelles, which was not recorded in the original report. Phylogenetic analyses were carried out based on SSU rRNA gene sequence data from each of the three species, and on other available data for microthoracids. The results showed that the order Microthoracida is not monophyletic because the family Discotrichidae, which contains L. paratorpens and D. papillifera, forms a clade separated from the 'core' Microthoracids clade. The topologies of the maximum likelihood and Bayesian inference trees, along with the distinct morphological characteristics found previously, suggest that the family Discotrichidae should not be assigned to the order Microthoracida. We propose to designate a new order, Discotrichida n. ord. which diagnosed as: flattened ciliates with conspicuous cortical papillae on both dorsal and ventral faces, rod-shaped mucocysts, and an asymmetric cytopharyngeal basket. Also, the fact that Leptopharynx clusters with the assemblage including T. australis, and Pseudomicrothorax is located distantly from Leptopharynx indicates that the classification of Pseudomicrothoracidae and Microthoracidae by Foissner (1985) is justified. [ABSTRACT FROM AUTHOR]

Extrusomes of different categories in ciliates have been suggested to have different structures and functions. To investigate the characteristics of extrusomes in the hypotrichous ciliate, we examinedArchitricha indicausing scanning and transmission electron microscopy and electron microscopic enzymo-cytochemistry. The results showed that the mature extrusomes were ellipsoidal vesicles with structureless cavities on their anterior parts and arranged in rows beneath the pellicle; the extruded structures of extrusomes were cyathiform and can remain outside of the cell surface after extrusion. These findings differ distinctly from that of previously reported extrusomes in other ciliates and thus suggest a new type of extrusomes. These organelles originated from endoplasmic reticulum in the deeper part of the cytoplasm, underwent a serious of development, gradually migrated to the cortical cytoplasm and finally positioned in their functional areas. In contrast with lower forms of ciliates, density, arrangement, and localization of extrusomes inA. indicashowed obvious differences between the ventral and dorsal surface of cells. This may be in some way related to the differentiation of the dorsoventral cortex, and functional ciliature happened to the hypotrichous ciliates. In addition, the extrusomes may perform protective or defensive function, which associates with the activity of mitochondria and contributes to the pellicle retrieval or renewal. [ABSTRACT FROM PUBLISHER]

Ciliature microtubular organelles, which organize into ciliary units, are highly complex microtubular systems. To investigate the formation of ciliature microtubular organelles and the role that different tubulins play in this process, we examined the distribution and function of α-, γ-, and δ-tubulins in the hypotrich ciliateStylonychia pustulata. Immunofluorescence analysis demonstrated that α-tubulin localized to all microtubular organelles, while γ- and δ-tubulins colocalized at basal bodies. Immunogold labeling revealed that γ- and δ-tubulins colocalize to the proximal part of basal bodies, suggesting that γ- and δ-tubulins probably interact and may function together. In addition, the permanent localization of γ-tubulin to the adoral zone of membranelles suggests that γ-tubulin play a role in the maintenance of oral ciliature base-associated microtubules, but does not play a role in the biogenesis of these ciliature. We observe that δ-tubulin is restricted to the basal body and the connective fibers of the basal body, suggesting this protein may be required for the extension and stability of the microtubules in these structures. Dramatically different concentrations of these three tubulins were observed in the cytoskeleton and the insoluble and soluble fractions of the cell, suggesting that these highly related proteins perform different functions inS. pustulata. [ABSTRACT FROM PUBLISHER]

Electron microscopic structure and enzymo-cytochemistry of light- and dark-cultured Paramecium bursaria was compared. In light-cultured Paramecium bursaria, the Chlorella symbionts grew normally and only a few of them were digested. In dark-cultured Paramecium bursaria, the number of food vacuoles containing symbionts increased significantly and the Chlorella symbionts in food vacuoles degenerated gradually and were finally digested by host cells. The activity of the enzymes acid phosphatase, adenosine triphosphatase, succinate dehydrogenase and glucose-6-phosphatase was studied in paramecia cultured in both light and dark conditions. The results showed that paramecia cultured under the two different conditions differed in their cytoplasmic structures and enzyme cytochemical reactions. The dark-cultured paramecia displayed stronger enzyme cytochemical reactivity. It is suggested that under dark-culture conditions, the gradual degeneration of Chlorella symbionts can stimulate their host to intensify its food uptake and the digestive activities of related organelles; whereas under the light-culture conditions, Chlorella symbionts can supply nutrition to their host and inhibit the functional activities of some organelles of their host. [Copyright &y& Elsevier]

Abstract: PRK2/PKNγ is a Rho effector and a member of the protein kinase C superfamily of serine/threonine kinases. Here, we explore the structure–function relationship between various motifs in the C-terminal half of PRK2 and its kinase activity and regulation. We report that two threonine residues at conserved phosphoacceptor position in the activation loop and the turn motif are essential for the catalytic activity of PRK2, but the phosphomimetic Asp-978 at hydrophobic motif is dispensable for kinase catalytic competence. Moreover, the PRK2-Δ958 mutant with the turn motif truncated still interacts with 3-phosphoinositide-dependent kinase-1 (PDK-1). Thus, both the intact hydrophobic motif and the turn motif in PRK2 are dispensable for the binding of PDK-1. We also found that while the last seven amino acid residues at the C-terminus of PRK2 are not required for the activation of the kinase by RhoA in vitro, however, the extreme C-terminal segment is critical for the full activation of PRK2 by RhoA in cells in a GTP-dependent manner. Our data suggest that the extreme C-terminus of PRK2 may represent a potential drug target for effector-specific pharmacological intervention of Rho-medicated biological processes. [Copyright &y& Elsevier]

Abstract: The classical swine fever virus (CSFV) full-length NS3 protein (NS3F) and the truncated NS3 protein (NS3H) with postulated helicase domain were expressed and demonstrated to have helicase activity. Further, the electrophoretic mobility shift assays containing NS3H and the viral 3′ terminal sequences showed that NS3H specifically bound to the plus- and minus-strand 3′UTR. The minus-strand 3′UTR had higher binding activity. The 21-nt fragments at the 3′-most terminal sequences of both 3′UTRs were essential to NS3H binding. A 12-nt insertion, CUUUUUUCUUUU, present in the 3′UTR of a CSFV live attenuated vaccine strain, was also found to be deleterious to helicase binding. Intact secondary structure of 3′ terminal sequence of 3′UTR might be important in helicase binding. Our results show that interaction between the helicase and the viral 3′UTR is similar to that between the replicase and the 3′UTR, suggesting that NS3 helicase is important for CSFV genomic replication. [Copyright &y& Elsevier]

The morphologies of two novel ciliates, Arcanisutura chongmingensis n. gen., n. sp. and Naxella paralucida n. sp., collected from Shanghai, China, have been investigated using live observation and silver staining methods. Arcanisutura n. gen. can be easily distinguished from related genera by its inconspicuous, oblique anterior suture. Arcanisutura chongmingensis n. sp. is mainly recognized by its elongated body with a tail-like posterior end, 25-33 somatic kineties, and 4-11 excretory pores. Naxella paralucida n. sp. can be distinguished from its congeners based on its two short nassulid organelles, fusiform trichocysts, 37-49 somatic kineties, and 16 nematodesmal rods. The small-subunit (SSU) rRNA gene sequences of these two species are presented, revealing the phylogenetic positions of Arcanisutura and Naxella. Phylogenetic analyses show that Arcanisutura forms a sister clade to other synhymeniid genera, namely, Chilodontopsis, Orthodonella, and Zosterodasys; Naxella is most closely related to Nassula spp. and is located within the monophyletic clade of the family Nassulidae., (© 2017 The Author(s) Journal of Eukaryotic Microbiology © 2017 International Society of Protistologists.)

Ciliates are able to form resting cysts as a survival strategy in response to stressful environmental factors. Studies on the characteristics of cellular structure during encystment may provide useful information for further understanding of the regulatory mechanism of cellular patterns and supply new clues regarding the phylogeny of ciliates. Scanning and transmission electron microscopies were used to observe the ultrastructure of cells during encystment of the soil ciliate Australocirrus cf. australis . The dedifferentiation of ciliature was revealed for the first time. Ciliary shafts first shortened, and the remaining ciliature, including basal bodies and the fibrillar cirral basket, retracted into the cytoplasm and was surrounded by the autophagic vacuoles and then gradually digested. A large number of autophagic vacuoles were observed in mature resting cysts. Autophagy might not only be necessary for the differentiation of cellular structures during encystment but might also be important to sustain the basic life activities in the resting stage. Australocirrus cf. australis formed a kinetosome-resorbing cyst and contained four layers in the cyst wall: the ectocyst, mesocyst, endocyst and granular layer. The ciliature resorbing state and the number of layers in the cyst wall were consistent with those found in other oxytrichous ciliates. However, the phenomenon wherein the two macronuclear nodules are not fused during encystment is not commonly observed among oxytrichids. Additionally, the octahedral granules in the mesocyst of this species exhibit different morphology from the congeners.

Cells of Cryptocaryon irritans at different life history stages were studied using both light and electron microscopy. The characteristics of several organelles were revealed for the first time at the ultrastructural level. It was confirmed that the cytostome of trophonts, protomonts and theronts was surrounded by cilium-palp triplets rather than ciliary triplets. The nematodesmata underlying the circumoral dikinetids were single bundles, whereas these were always paired in Prorodontids. Toxicysts were present in late-stage tomonts and theronts, but were absent in trophonts and protomonts. We posited that toxicysts might play a role in infection and invasion of host-fish tissue by theronts. The adoral brosse was unlike that of any other family of the class Prostomatea based on its location and morphology. Membranous folds were present in trophonts, protomonts and theronts. These folds were longer and more highly developed in C. irritans than in exclusively free-living prostome ciliates suggesting that they might be linked to parasitism in C. irritans. Trophonts, protomonts and theronts had multiple contractile vacuoles. The basic ultrastructure of the contractile vacuole of C. irritans was similar to that of other kinetofragminophoran ciliates. They might play different roles in different stages of the life cycle since their ultrastructure varied among trophonts, protomonts and theronts.

The morphology and infraciliature of the four hypotrichous ciliates; Rigidohymena inquieta (Stokes, 1887) Berger, 2011, Pattersoniella vitiphila Foissner, 1987, Notohymena australis Foissner & O' Donoghue, 1990, and Cyrtohymena (Cyrtohymenides) australis (Foissner, 1995) Foissner, 2004, collected from east China, were investigated by using live observation and protargol impregnation method. An improved diagnosis for R. inquieta was supplied based on descriptions of present and previous populations. New morphology and morphogenesis information based on Chinese populations of another three hypotrichids were also supplemented. The Small-subunit rRNA (SSU rRNA) gene sequences of the four species were characterized and their phylogenetic positions were revealed by means of Bayesian inference and Maximum-likelihood analysis. The analyses shows that R. inquieta clusters with other members of the subfamily Stylonychinae, which confirms the monophyly of the subfamily and verified R. inquieta as a separated species from R. candens though it differs from others mainly by body size. C. (C.) australis occupying the basal position of the clade which contains cyrtohymenids and some other groups, declines the idea of separating Cyrtohymena into two subgenus. Notohymena australis and China population of Pattersoniella vitiphila respectively clustering with their congeners correspond well with the systematics revealed by morphological similarities.

The morphology and infraciliature of two soil ciliates, Parafurgasonia zhangi spec. nov. and Chilodonella acuta Kahl, 1931, collected from Saudi Arabia, were investigated by observations of both living cells and specimens after standard staining methods. P. zhangi differs from its congeners by the combination of the following features: excretory pore quite near posterior end of paroral membrane, 16 or 17 somatic kineties with about 11 kinetids in each one on dorsal side, paroral membrane gently curved and composed of about 15 dikinetids, and hypostomial organelle composed of four or five files of kinetids with four monokinetids each. The diagnosis of Chilodonella acuta was renewed to include characteristics revealed by the silver impregnation method: cells in vivo measuring 33-45) × 18-26) µm, dorsal hump and tail-like podite present, two contractile vacuoles, seven left and five right kineties, 9-11 nematodesmal rods, and dorsal brush containing about 11 basal bodies. Phylogenetic analyses based on small-subunit rRNA gene sequences showed that P. zhangi was closer to species of the Colpodidiidae rather than the Furgasoniidae represented by Furgasonia blochmanni, and Chilodonella acuta clustered with its congener Chilodonella uncinata but was a well-outlined species of the genus., (© 2014 IUMS.)

Two Dexiotricha species (Dexiotricha elliptica nov. comb. and Dexiotricha cf. granulosa), respectively isolated from soil north-west of Riyadh, Saudi Arabia, and freshwater in Shanghai, eastern China, were investigated using standard methods. The species Loxocephalus ellipticus Kahl, 1931 is reclassified here in the genus Dexiotricha and was characterized mainly by constantly showing 16 somatic kineties, three post-oral kineties with the middle one shortened, a contractile vacuole located subcaudally with an excretory pore near the posterior end of somatic kinety 2 and single caudal cilia. A Dexiotricha granulosa-like organism having a subcaudally located contractile vacuole and fewer somatic kineties was designated D. cf. granulosa. The small-subunit rRNA gene (SSU rDNA) sequences of these two species were characterized and their phylogenetic positions based on SSU rDNA sequences were revealed by means of Bayesian inference and maximum-likelihood analysis. Phylogenetic analyses confirmed Dexiotricha as a monophyletic genus and supported its assignment to the order Loxocephalida. However, its family assignment remains unsupported.

Three species of Microthoracids, Lopezoterenia paratorpens n. sp., Trochiliopsis australis Foissner et al., 1988 and Discotricha papillifera Tuffrau, 1954, collected from Chinese coastal waters, were investigated using live observation and protargol staining methods. Lopezoterenia paratorpens n. sp. was characterized by its squarely shaped cortical papillae and by dorsal kineties which contained loosely distributed basal bodies. Trochiliopsis australis was revealed to have two oral membranelles, which was not recorded in the original report. Phylogenetic analyses were carried out based on SSU rRNA gene sequence data from each of the three species, and on other available data for microthoracids. The results showed that the order Microthoracida is not monophyletic because the family Discotrichidae, which contains L. paratorpens and D. papillifera, forms a clade separated from the "core" Microthoracids clade. The topologies of the ML and BI trees, along with the distinct morphological characteristics found previously, suggest that the family Discotrichidae should not be assigned to the order Microthoracida. We propose to designate a new order, Discotrichida n. ord. which diagnosed as: flattened ciliates with conspicuous cortical papillae on both dorsal and ventral faces, rod-shaped mucocysts, and an asymmetric cytopharyngeal basket. Also, the fact that Leptopharynx clusters with the assemblage including T. australis, and Pseudomicrothorax is located distantly from Leptopharynx indicates that the classification of Pseudomicrothoracidae and Microthoracidae by Foissner (1985) is justified. This article is protected by copyright. All rights reserved., (This article is protected by copyright. All rights reserved.)