Two new scuticociliates from southern China: Uronema apomarinum sp. nov. and Homalogastra parasetosa sp. nov., with improved diagnoses of the genus Homalogastra and its type species Homalogastra setosa (Ciliophora, Oligohymenophorea).

The morphology of two new scuticociliates, Uronema apomarinum sp. nov. and Homalogastra parasetosa sp. nov., isolated from a mangrove wetland in Shenzhen, PR China, was studied using live observation and the protargol impregnation method. Uronema apomarinum is characterized by a body size of about 20-35×10-15 µm in vivo, a partly two-rowed membranelle 1, and 12 or 13 somatic kineties. Homalogastra parasetosa is distinguished by a membranelle 1 comprising two longitudinal rows of basal bodies. Three Homalogastra setosa populations are suggested as subjective synonyms of the new species. Improved diagnoses of the genus Homalogastra Kahl, 1926 and its type species Homalogastra setosa Kahl, 1926 are provided. Results of phylogenetic analyses based on 18S rRNA gene and ITS1-5.8S-ITS2 region sequences indicate that U. apomarinum is most closely related to U. marinum, while the closest relative of H. parasetosa is H. setosa.

Morphological redescriptions and neotypification of two poorly known tintinnine ciliates (Alveolata, Ciliophora, Tintinnina), with a phylogenetic investigation based on SSU rRNA gene sequences.

Two poorly known tintinnine ciliates collected from the coastal waters of PR China, viz., Codonellopsis mobilis Wang, 1936 and Tintinnopsis chinglanensis Nie & Ch'eng, 1947, were redescribed and neotypified using live observation, protargol staining and SSU rRNA gene sequencing. Ciliature information and SSU rRNA gene sequence data of both species were revealed for the first time and improved diagnoses were given based on the original descriptions and data from the present study. Further phylogenetic analyses inferred from SSU rRNA gene sequences and morphological data suggested that the genus Tintinnopsis is polyphyletic and that the genus Codonellopsis is non-monophyletic. The approximately unbiased test, however, does not reject the possibility that Codonellopsis is monophyletic.

Analysis of silver-associated proteins in pathogen via combination of native SDS-PAGE, fluorescent staining, and inductively coupled plasma mass spectrometry.

Characterization of silver-associated proteins is important to elucidate underlined mechanisms of silver-containing materials against microbes. Gel electrophoresis based methods are the most popular and basic strategy for the analysis of biomolecules, i.e., proteins and nucleic acids. It solely provides molecular weights of analytes. Extending the method from molecular weight to elemental composition is highly desired when investigating metal-containing molecules. Herein, a gel electrophoresis based method combining native sodium dodecyl sulfate-polyacrylamide gel electrophoresis (native SDS-PAGE), fluorescent staining, and inductively coupled plasma mass spectrometry (ICP-MS) strategy was developed for separation and detection of silver-associated proteins. Two home-made silver-labeled proteins, carbonic anhydrase and ovalbumin, were used for validation of the strategy performance. Silver-associated proteins in Pseudomonas aeruginosa and Staphylococcus aureus treated with silver nanoparticles were further characterized by this method. Some well-known and new proteins were identified to associate to silver in both P. aeruginosa and S. aureus, demonstrating the feasibility of the developed strategy. In conclusion, the current study provides a convenient method for readily identification of silver-associated proteins in biological samples.

An Optimized Protocol of Protargol Staining for Ciliated Protozoa.

Protargol staining is a crucial method to reveal the infraciliature of ciliates, which is the most important morphological character for species identification. In the present study, Wilbert's protocol of protargol staining was emended mainly toward the highly happened improper bleaching. Through reciprocal treatments, both insufficient and excessive bleachings were much eliminated from the protargol protocol and the tests performed with four different species of ciliates established that the stainings were considerably improved and more reliable with optimized bleaching. Compared to the original protocol, the optimized method was proved to be more suitable for the groups difficult to stain, and it is also friendlier for the beginners and researchers in related fields.

Dartintinnus alderae n. g., n. sp., a Brackish Water Tintinnid (Ciliophora, Spirotrichea) with Dual-ended Lorica Collapsibility.

A tintinnid ciliate isolated from waters of the Thames River (Connecticut, USA) is described through combined in vivo observation, protargol impregnation, and phylogenetic analysis. The novel genus Dartintinnus and its type species, D. alderae are distinct from established tintinnid taxa by a lorica that collapses on both anterior and posterior ends. Dartintinnus is placed in the family Eutintinnidae based on a hyaline, elongated lorica opened at both ends, a ciliary pattern including a ventral kinety, at least one dorsal kinety, and right, left and lateral fields, and a sister relationship with Eutintinnus in gene trees. Main differences between D. alderae and Eutintinnus species include a 5.5-6.5% divergence in the small subunit rRNA gene, the geometry of the lorica (resembling an isosceles tetrahedron when collapsed vs. a cylinder, respectively), the number of macronuclear nodules (two vs. four), and the number of dorsal kineties (one vs. usually two). Considering the features of the new genus, we improve the diagnosis of the family Eutintinnidae, including the presence of a lateral ciliary field that had been overlooked in some Eutintinnus species. This work exemplifies the potential for novel diversity, even in these relatively well-studied protists, and the importance of an integrated approach for the description of tintinnid taxa.

A cobertura ideal para tratamento em paciente queimado: uma revisão integrativa da literatura / The ideal coverage for treatment in a burning patient: an integrating literature review / La cobertura ideal para el tratamento en un paciente quemado: una revisión de la literatura integrando

Objetivo: O presente estudo objetiva uma revisão de literatura detalhada a respeito da difícil escolha da melhor cobertura do paciente acometido por queimaduras. Método: A análise foi realizada por meio de pesquisa de literatura on-line em periódicos nacionais e internacionais relacionados com o tema. Tratase de uma revisão integrativa da literatura, na qual a coleta de dados ocorreu no período de agosto de 2016 a agosto de 2017. As bases de dados utilizadas foram: MEDLINE (Medical Literature Analysis and Retrieval System Online), Literatura Latino-Americana e do Caribe em Ciências da Saúde (LILACS), Scientific Eletronic Library Online (Scielo), por meio de busca na Biblioteca Virtual em Saúde (BVS). Foi estabelecido o recorte do tempo para inclusão de publicações de 2006 a 2017. Resultados: Existem vários tipos de coberturas indicadas no tratamento da queimadura. A prata é o tratamento padrão. Nos dias atuais, diante das várias opções e inovações, tem-se utilizado prata mais associações, devido à eficácia e melhor custo-benefício. Conclusão: É necessário conhecimento específico do dermatoterapeuta para que o tratamento ao paciente queimado tenha êxito. É preciso treinar enfermeiros para oferecer o suporte necessário a esses pacientes, atendendo-os de forma ímpar, com estudo específico de cada caso. Sugere-se a prata com associações para o tratamento da queimadura. A literatura sobre esse tema é escassa. Sendo assim, o surgimento de novas terapias mais eficazes ainda é esperado.

Objective: The present study aims at a detailed literature review regarding the difficult choice of the best coverage of patients suffering from burns, considering a range of options. Methods: The analysis was performed through online literature research in national and international journals related to the topic. It is an integrative review of the literature, in which the data collection took place from August 2016 to August 2017. The databases used were: MEDLINE (Medical Literature Analysis and Retrieval System Online), Latin American Literature and the Caribbean in Health Sciences (LILACS), Scientific Electronic Library Online (Scielo), through the search in the Virtual Health Library (VHL). The time cut was established for the inclusion of publications from 2006 to 2017. Results: There are several types of coverage indicated in the treatment of burn. Silver is the standard treatment. Nowadays, in front of the various options and innovations has been used more silver associations, due to the effectiveness and better cost-benefit. Conclusion: Specific knowledge of the dermatologist is necessary so that the treatment of the burned patient can be successfully achieved. It is necessary to train nurses to offer the necessary support to these patients, attending them in an odd way, with specific study of each case. Silver is suggested with associations for the treatment of burn. The literature on this subject is scarce. Thus, the emergence of more effective new therapies is still expected

Objetivo: El presente estudio objetiva una revisión de literatura detallada acerca de la difícil elección, de la mejor cobertura del paciente acometida por quemaduras. Método: El análisis fue realizado a través de la investigación de literatura on line en periódicos nacionales e internacionales relacionados con el tema. Se trata de una revisión integrativa de la literatura, en la cual la recolección de datos ocurrió en el período de agosto de 2016 a agosto de 2017. Las bases de datos utilizadas fueron: MEDLINE (Medical Literature Analysis and Retrieval System Online), Literatura Latinoamericana y el Caribe en Ciencias de la Salud (LILACS), Scientific Eletronic Library Online (Scielo), a través de la búsqueda en la Biblioteca Virtual en Salud (BVS). Se estableció el recorte del tiempo para la inclusión de publicaciones de 2006 a 2017. Resultados: Existen varios tipos de coberturas indicadas en el tratamiento de la quemadura. La plata es el tratamiento estándar. En los días actuales, frente a las diversas opciones e innovaciones se han utilizado plata más asociaciones, debido a la eficacia y mejor costo-beneficio. Conclusión: Se necesita conocimiento específico del dermatoterapeuta para que el tratamiento al paciente quemado pueda ser logrado con éxito. Es necesario entrenar a enfermeros para ofrecer el soporte necesario a estos pacientes, atendiéndolos de forma impar, con estudio específico de cada caso. Se sugiere la plata con las asociacio

Morphogenesis of a saline soil ciliate Urosoma salmastra (Dragesco and Dragesco-Kernéis, 1986) Berger, 1999 with notes on the phylogeny of Urosoma (Ciliophora, Hypotrichia).

Morphogenesis in a population of Urosoma salmastra collected from saline soil in Weinan, China, was investigated using protargol staining. The main morphogenetic event that characterizes U. salmastra is the inclusion of cirrus III/2 in the formation of the frontoventra-transverse cirral anlagen. We also provide small subunit ribosomal DNA gene sequences for a population of U. salmastra and for two populations of U. emarginata. The molecular phylogeny indicates that Urosoma is not monophyletic, viz, U. emarginata branches separately from its congeners and the closest relative of U. salmastra is Oxytricha granulifera.

Redescription and Phylogenetic Position of Condylostoma arenarium Spiegel, 1926 (Ciliophora, Heterotrichea) from Guanabara Bay, Brazil.

Details on Condylostoma arenarium infraciliature have not been described; therefore, it is considered a poorly known species. The lack of detailed description on C. arenarium morphology caused several misidentifications that have accumulated in the literature. In this study, we present the first complete description of C. arenarium infraciliature based on protargol-impregnated organisms and scanning electron microscopy. We also have inferred the phylogenetic position of this species based on 18S rRNA sequences. The main characteristics of C. arenarium population from Guanabara Bay are as follows: in vivo elongated body shape with 350-600 µm length × 70-220 µm width, they are highly contractile when subjected to disturbances, green-yellowish cortical granules are present, contractile vacuoles absent, V-shaped peristome comprises approximately 1/5 of the total length, adoral zone with 83-145 membranelles, 1-2 small frontal cirrus observed only in impregnated specimens, 10-15 fiber-like stripes arranged transversely on the inner wall of the oral cavity, 30-45 somatic kineties, moniliform macronucleus with 15-20 nodules. Some observations on morphogenesis of C. arenarium were also included. In phylogenetic analyses, C. arenarium clustered with Condylostoma sp. within a clade composed of three C. curva sequences with high support values.

Phylogenetic position of the genus Cyrtostrombidium, with a description of Cyrtostrombidium paralongisomum nov. spec. and a redescription of Cyrtostrombidium longisomum Lynn & Gilron, 1993 (Protozoa, Ciliophora) based on live observation, protargol impregnation, and 18S rDNA sequences.

We redescribe Cyrtostrombidium longisomum Lynn & Gilron, 1993, the type species of the genus Cyrtostrombidium, and describe the new species Cyrtostrombidium paralongisomum n. sp. using live observation, protargol staining and molecular data. The morphological characters of these two species are clearly distinct, i.e., dikinetid numbers in the girdle and ventral kineties; however, it is difficult to separate them by 18S rDNA sequences because they differ by only 8 bp, indicating that 18S rDNA sequences are insufficient for separating different species in the genus Cyrtostrombidium. We not only observed the position of the oral primordium in the genus Cyrtostrombidium but also observed a possibly homoplasious trait, a dorsal split in the girdle kinety, in (1) Apostrombidium, (2) Varistrombidium, and (3) Cyrtostrombidium/Williophrya. This partially supports the hypothesis of somatic ciliary pattern evolution recently put forth by Agatha and Strüder-Kypke.

[The comparative effectiveness of framycetin included in combined therapy of adenoiditis in the children].

The objective of the present study was to evaluate the effectiveness of framycetin included in combined therapy of adenoiditis in the children. The study involved 67 children at the mean age of 6.9±2.7 years. Group 1 was comprised of 35 children given framycetin as topical therapy, the patients of group 2 were treated by the endonasal administration of a 2% silver proteinate solution. It was shown that the use of framycetin as a component of combined therapy of adenoiditis enhances the effectiveness of the treatment and compliance to therapy in comparison with the same parameters in the case of the application of traditional topical antibacterial preparations.

An update of 'basic light and scanning electron microscopic methods for taxonomic studies of ciliated protozoa'.

This is an update of the review by Foissner (1991). Since then, I have improved some methods considerably. The following methods are described in detail: live observation, supravital staining with methyl green-pyronin, dry silver nitrate impregnation, wet silver nitrate impregnation, silver carbonate impregnation, protargol impregnation (three procedures), scanning electron microscopy, and deciliation. Familiarity with these methods (or modifications) is a prerequisite for successful taxonomic work. No staining method is equally appropriate to all kinds of ciliates. A table is provided which indicates those procedures which work best for certain groups of ciliates. A second table relates to the structures revealed by the procedures. Good descriptions usually demand at least live observation, silver nitrate and protargol or silver carbonate impregnation. Some instructions are provided for distinguishing mono- and dikinetids as well as ciliated and non-ciliated basal bodies in silvered ciliates. Furthermore, I added a section on 'Deposition and Labeling of Preparations'. All methods work not only with ciliates but also with many other heterotrophic and autotrophic flagellated and amoeboid protists. The brilliancy of silver preparations has tempted some taxonomists to neglect live observation. However, many important species characteristics cannot be seen or are changed in silvered specimens. I thus consider all species descriptions based exclusively on silver slides as incomplete and of doubtful value for both α-taxonomists and ecologists.

Technique: imaging earliest tooth development in 3D using a silver-based tissue contrast agent.

Looking in microscopic detail at the 3D organization of initiating teeth within the embryonic jaw has long-proved technologically challenging because of the radio-translucency of these tiny un-mineralized oral tissues. Yet 3D image data showing changes in the physical relationships among developing tooth and jaw tissues are vital to understand the coordinated morphogenesis of vertebrate teeth and jaws as an animal grows and as species evolve. Here, we present a new synchrotron-based scanning solution to image odontogenesis in 3D and in histological detail using a silver-based contrast agent. We stained fixed, intact wild-type mice aged embryonic (E) day 10 to birth with 1% Protargol-S at 37°C for 12-32 hr. Specimens were scanned at 4-10 µm pixel size at 28 keV, just above the silver K-edge, using micro-computed tomography (µCT) at the Canadian Light Source synchrotron. Synchrotron µCT scans of silver-stained embryos showed even the earliest visible stages of tooth initiation, as well as many other tissue types and structures, in histological detail. Silver stain penetration was optimal for imaging structures in intact embryos E15 and younger. This silver stain method offers a powerful yet straightforward approach to visualize at high-resolution and in 3D the earliest stages of odontogenesis in situ, and demonstrates the important of studying the tooth organ in all three planes of view.

Protargol synthesis: an in-house protocol.

The protargol staining method has proved to be indispensable for revealing the cellular structures of a variety of protozoa, especially the flagellates and ciliates. Protargol provides permanent stains of a variety of cellular structures: nuclei, extrusomes, basal bodies, and microfilamentous constituents of cells. Together with the older silver nitrate methods, protargol impregnations have provided the basis for the detailed descriptions of nearly all ciliates to date. The performance of commercially available preparations has varied widely. Recently, suppliers have stopped stocking the effective forms of protargol resulting in a worldwide shortage. Thus, it has become necessary for protistologists to explore on-site synthesis of this critically important agent. An optimum protocol for synthesis of protargol should be rapid, relatively inexpensive, simple enough to be done by non-chemists, and achievable without specialized equipment. In this article, the authors briefly review the interesting history of protargol and describe a protocol, based on the early studies of neuroanatomists, that yields a protargol producing impregnations of ciliates comparable to those obtained with previously available commercial preparations.

New observations on the ciliate genus Vestibulongum (Pycnotrichidae): vestibular ultrastructure, macronuclear endosymbiotic bacteria, biogeography, and evidence for host specificity.

Two isolates of the pycnotrichid ciliate genus, Vestibulongum, were collected from the host fish, Acanthurus xanthopterus, from two locations in the Southern Pacific Ocean. One was from the Great Barrier Reef (GBR), and a second from Papua New Guinea. These sites are thousands of km from the type locality, off the coast of South Africa. New data were collected from protargol-stained samples to more fully characterize the general form and light microscopic structures of the ciliate. Specimens from all three sites had a long vestibule, characteristic of most members of the family. Data suggest that specimens from each site are the same genus. The kinetids of the Vestibulongum isolated from the GBR contained the typical components of postciliary, transverse, and nemodesmatal microtubules, and Kd fibrils. Also, two quite different forms of endomacronuclear bacteria were observed and are described. One of those has distinct endospores, which are similar to endospores in nuclear endosymbiotic bacteria in a species of Balantidium from the gut of another species of surgeonfish.

Three new Loxophyllum species (Ciliophora: Pleurostomatida) from China with a brief review of the marine and brackish Loxophyllum species.

Recent studies indicate that there is a high diversity of pleurostomatid ciliates in the coastal waters of China. Here, three new congeners of Loxophyllum, L. caudatum sp. n., L. rugosum sp. n., and L. chinense sp. n., are described following observations of live cells and protargol-impregnated specimens. All three species usually have two macronuclear nodules and prominent warts along the dorsal margin formed by clustered extrusomes. In addition, L. caudatum sp. n. is characterized by its long conspicuous tail, dot-like cortical granules, 4 or 5 left and 9 or 10 right kineties, and a single subterminal contractile vacuole. Loxophyllum rugosum sp. n. is distinguished by possessing three prominent ridges on the left side, 7-11 right and 5-7 left kineties. Loxophyllum chinense sp. n. is characterized by having several contractile vacuoles distributed along the ventral margin, 13-18 right and 6-8 left kineties. The small subunit ribosomal DNA (SSU rDNA) sequence similarities among six congeners range from 96.46% to 99.94%. Phylogenetic trees based on the SSU rDNA sequences indicate that all Loxophyllum spp. form a well-supported monophyletic group. A brief review of the marine and brackish Loxophyllum species is supplied and one new combination, Litonotus multiplicatus (Kahl 1931) comb. n. (basionym Loxophyllum multiplicatum Kahl 1931), and one new name, Litonotus dragescoi nom. n. (basionym L. fasciolatus Dragesco 1966), are suggested.

A new interpretation of stomatogenesis in a peritrich ciliate: using Campanella umbellaria as a model system.

The process of stomatogenesis in peritrich ciliates is still incompletely understood. Previous studies on the stomatogenesis of four species of peritrichs, Telotrochidium sp., Carchesium polypinum, Opercularia coarctata, and Astylozoon pyriforme conflict with one another in some cases and omit details of events in others. We described the entire process of stomatogenesis in the peritrich ciliate Campanella umbellaria (C. umbellaria) using an improved method of staining with protargol. Our results disagree with some previous studies with regard to the formation of some rudimentary structures, reorganization of the parental haplokinety, formation of new germinal rows, and separation of daughter oral complexes. The pattern of stomatogenesis characteristic of peritrichs is compared to the stomatogenetic patterns of three other oligohymenophorean subclasses and a hypothesis about the evolution of stomatogenesis in the class Oligohymenophorea is offered. Details of stomatogenesis need to be described and verified in a greater variety of peritrichs to clarify possible differences between taxa and make it possible to relate stomatogenesis to evolution within the subclass Peritrichia. Ultrastructural studies are the next step in description of morphogenetic processes in peritrichs, and characteristics of C. umbellaria make it a useful model for this work.

Morphological redescriptions of four marine ciliates (Ciliophora: Cyrtophorida: Dysteriidae) from Qingdao, China.

The morphology and infraciliature of four marine cyrtophorid ciliates isolated from Qingdao, China, were investigated. Based on the present work and on previous data, improved diagnoses for three rarely known species are provided: (1) Mirodysteria decora; small-sized marine Mirodysteria about 35-60 × 25-35 µm in vivo, oval in outline; body surface with two or three conspicuous dorsal spines and one caudal spine; three right kineties, the rightmost one extending dorso-apically; left frontal kineties reduced, each consisting of three basal bodies only; podite subcaudally positioned; two ventrally located contractile vacuoles. (2) Dysteria legumen; body oval with two longitudinal grooves on different plates; six right kineties, the rightmost two of which extend dorso-apically; two left frontal kineties and two ventrally located contractile vacuoles. (3) Dysteria proraefrons; body about 60×35 µm in vivo; six right kineties, the two rightmost of which extend dorso-apically and the leftmost one is considerably shortened; three left frontal kineties; two ventrally located contractile vacuoles. A population of D. derouxi with eight or nine right kineties is also briefly described. The current investigation further demonstrates high diversity and cosmopolitan distribution of this highly specialized group of benthic ciliates.

Morphological and molecular information of a new species of Geleia (Ciliophora, Karyorelictea), with redescriptions of two Kentrophoros species from China.

The morphology and infraciliature of three karyorelictean ciliates, Geleia sinica spec. nov. and two poorly known Kentrophoros species, K. flavus and K. gracilis, isolated from the intertidal zone of a beach at Qingdao, China, were investigated. Geleia sinica spec. nov. is distinguished from its congeners by the following combination of characters: body medium-sized and slender-cylindrical; with a conspicuous prebuccal fossa; 28-34 somatic kineties; about 40 short adoral polykineties; intrabuccal kinety composed of 25-34 dikinetids; paroral kineties composed of closely spaced dikinetids. The comparison with similar congeners clearly supports the validity of this new species based on morphological and small subunit (SSU) rRNA gene sequence data. In light of these new data the "well-known" morphotype, Geleia simplex (Fauré-Fremiet, 1951), is redefined. Two Kentrophoros species are redescribed and improved diagnoses are supplied. Kentrophoros flavus Raikov and Kovaleva, 1968 is mainly characterized by having about 33 macronuclei and 12 micronuclei forming a row that extends along the cell meridian, and 12-19 ciliary rows on the right side of the cell. Kentrophoros gracilis Raikov, 1963 is characterized by having about 14 macronuclei, 13 micronuclei and 10-13 kineties on the right side of the cell.

Morphological description of three marine ciliates (Ciliophora, Scuticociliatia), with establishment of a new genus and two new species.

Three marine scuticociliates, Falcicyclidium fangi nov. gen., nov. spec., Falcicyclidium atractodes nov. spec., and Cristigera media Kahl, 1928 were investigated using live observation and silver impregnation methods. The genus Falcicyclidium is distinguished by the combination of: (i) dorsoventrally flattened body, (ii) hook-like (falciform) paroral membrane, (iii) anterior end of paroral membrane posterior to anterior end of membranelle 1, and (iv) multiple caudal cilia. Falcicyclidium fangi nov. spec., the type of the new genus, can be recognized by the combination of its large size, extremely dorsoventrally flattened (3:1) body, consistently 10 somatic kineties, and the broad, elongate buccal area occupying 60% of the body length. Falcicyclidium atractodes nov. spec. is mainly characterized by a unique spine projecting from both the anterior and posterior end. The uncommon form, Cristigera media is redescribed based on the population from Qingdao, the statistic data and additional features, especially the morphology of the living cells, are documented.