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Biological soil crusts (BSC) are associations of different macro and microorganisms and aggregated soil particles located on the surface of soils in many different habitats. BSC harbour a diverse and complex community of ciliates and testate amoebae. These phagotrophic protists play an important role in C and N recycling in soil ecosystems but have not been frequently studied in BSC. In this context, the effects of three increasing N inputs on ciliates and testate amoebae in crusts from a semi-arid Mediterranean ecosystem were evaluated. A field experiment with artificial N-deposition was designed to mimic the effects caused by anthropogenic N depositions. The results have shown that the protist populations of these semi-arid Mediterranean environments have lower species richness than other soil environments. The increase in N produces a net loss of diversity in the populations studied and shifts in the community structure. It has also been shown that some ciliates and testate amoebae, due to their population responses to increased N concentrations, could potentially be used as bio-indicators of N contamination in these BSCs.

Rain-fed granite rock pools are geological landforms with a worldwide distribution. However, their role as habitats for microorganisms has been barely explored. We carried out a detailed morphological inventory of the ciliated protists in the sediments of three granite rain-fed rock pools from a Spanish National Park. The ciliate seed bank in the rock pools comprised 54 morphospecies. The species number inferred for each pool by rarefaction analysis was similar to that observed by microscopy. The most representative species were small bacterivorous ciliates, although the distribution of ciliate groups was significantly different in each rock pool. Testate amoebae were also found to be rich in species. This study demonstrates the existence of a diverse ciliate community adapted to persist in these ephemeral and extreme rock pool habitats through species resistance structures, many of which have not been described previously. The presence of competitive species in dormant stages prevents local extinctions through a sequential excystation over time. Our results provide a benchmark for studying protist and microbial biodiversity within rain-fed granite rock pools, a unique habitat that merits bio- and geo- conservation.

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.

Las pilas representan un tipo de forma presente en casi todos los ambientes climáticos. El trabajo estudia el papel de los ciclos térmicos como un agente importante del modelado y asimetría que presentan las paredes de las pilas en un clima Mediterrá- neo templado - frío. El estudio se ha realizado en el macizo granítico de la Pedriza de Manzanares, zona protegida de gran valor ambiental, incluida en el Parque Nacional de la Sierra de Guadarrama (Madrid, España). El análisis de la variabilidad térmica mediante modelos de regresión perió- dica múltiple, pone de manifiesto la influencia de los ciclos diario y anual en función de las orientaciones norte y sur de las paredes de las pilas. Un modelo matemático de regresión lineal muestra cómo la variabilidad térmica diaria influye en la alteración de las paredes de las pilas, generando superficies planas o de concavidad más o menos pronunciada. El trabajo también establece relaciones entre el grado de alteración de las diferentes partes de las pilas, con la presencia de líquenes y dureza relativa de la roca. Los resultados avalan la hipótesis de relación causal entre la variabilidad térmica y alteración de las paredes de las pilas según su orientación norte o sur.

Unusual prostomatid specimens were found in the biological reactor of a wastewater treatment plant in a health resort in Valencia, Spain. These ciliates were attached to flocs unlike other free-swimming prostomatid ciliates described to date in the mixed liquor of activated sludge plants. The morphological study of this species led to a typically different combination of characteristics: elongated cell shape, 20-30 somatic kineties, 2 perioral kineties, and 1 circumoral kinety, 1 large vacuole protruding at the terminal end, a lorica tapered toward the aperture with a smooth neck, and 11-16 annular ridges. These characteristics place this representative as a new species of the genus Metacystis-Metacystis galiani n. sp. This species became the dominant population within the biological reactor when high values of conductivity (4,244 mS/cm) and temperature (26.8 degrees C) were recorded.

International audience; Since ciliates rarely possess structures that easily fossilize, we are limited in our ability to use paleontological studies to reconstruct the early evolution of this large and ecologically important clade of protists. Tintinnids, a group of loricate (house-forming) planktonic ciliates, are the only group that has a significant fossil record. Putative tintinnid fossils from rocks older than Jurassic, however, possess few to no characters that can be found in extant ciliates; these fossils are best described as `incertae sedis eukaryotes'. Here, we review the Devonian fossil Nassacysta reticulata and propose that it is likewise another `incertae sedis eukaryote due to the lack of any unambiguous ciliate characters. Future tintinnid fossil descriptions would be most helpful if: (i) neutral terminology is used in the descriptions but ciliate-specific terminology in the interpretations; (ii) the current ciliate classification is used, although fossil data may expand or modify classifications based on modem forms; (iii) close collaboration with specialists studying extant ciliates is done; and (iv) editors include an expert of extant ciliates in the review process. (C) 2015 Elsevier B.V. All rights reserved.

We give an account of the loricate mixotrophic nanoflagellate Ollicola vangoorii (basionym Codonomonas Van Goorii, synonym Calycomonas Van Goorii) (Chrysomonadidae = Chrysophyceae p.p. ) in the plankton of the western Mediterranean Sea. Using digital imaging and field-emission scanning electron microscopy (SEM), we describe the lorica morphology and ultrastructure based on some 150 specimens occurring either as dispersed (unattached) cells or cells attached to suspended particulate matter in coastal waters off Barcelona (Catalonia, Spain) in July 1998. The spectrum of morphological variation includes that of Calycomonas gracilis sensu Wulff non Lohmann, Calycomonas gracilis sensu Espeland & Throndsen non Lohmann, Calycomonas wulffii Conrad et Kufferath, Ollicola cylindrical (Conrad et Kufferath) Vørs (= Calycomonas cylindrical (Conrad et Kufferath) Lund, Codonomonas cylindrica Conrad et Kufferath), Ollicola dilatata (Conrad et Kufferath) Vørs (= Calycomonas dilatata (Conrad et Kufferath) Lund, Codonomonas dilatata Conrad et Kufferath), suggesting that those taxa are conspecific with Ollicola vangoorii. By contrast, Ollicola pascheri (van Goor) Vørs (basionym Codonomonas pascheri van Goor) is best maintained as a separate species and retained in the genus Codonomonas van Goor. From an ultrastructural viewpoint, the basic architecture of the lorica is that of a fibrillar ribbon coiled around the cell and eventually hardened as a result of inorganic mineralization from the environment. The appearance of the fibrillar component and the degree of mineralization may vary considerably. The cell apex is modified in a newly documented apical cytostome, substantiating an existing observation that cells are capable of ingesting bacteria. Although the statistical correlation between SEM-based nanoflagellate cell counts and epifluorescence-based bacterial cell counts is at best only weakly significant, it is possible that in the Barcelona populations O. vangoorii relies partly on bacterivory and therefore its ecological role is partly that of a carbon consumer. We also hypothesize that the presence of an external lorica increases the probability of intercepting bacterial prey, ready to be ingested by the cell in the apical cytostome. Aportamos una descripción del nanoflagelado mixotrófico provisto de lorica Ollicola vangoorii (basiónimo Codonomonas Van Goorii, sinónimo Calycomonas Van Goorii) (Chrysomonadidae = Chrysophyceae p.p.) en el plancton del Mar Mediterráneo Noroccidental. Mediante la utilización de imágenes digitalizadas y un microscopio electrónico de barrido (SEM), describimos la morfología de la lórica y la ultraestructura basándonos en 150 especímenes que aparecían ya fueran en células dispersas (no adheridas) ó células adheridas a material particulado suspendido en aguas costeras a poca distancia de Barcelona (Catalunya, España) en Julio de 1998. El espectro morfológico incluye a Calycomonas gracilis sensu Wulff non Lohmann, Calycomonas gracilis sensu Espeland & Throndsen non Lohmann, Calycomonas wulffii Conrad et Kufferath, Olli cola cylindrica (Conrad et Kufferath) Vørs (= Calycomonas cylindrica (Conrad et Kufferath) Lund, Codonomonas cylindrica Conrad et Kufferath), Ollicola dilatata (Conrad et Kufferath) Vørs (= Calycomonas dilatata (Conrad et Kufferath) Lund, Codonomonas dilatata Conrad et Kufferath), sugiriendo que aquellos taxones eran conespecíficos con Ollicola vangoorii. En cambio Ollicola pascheri (van Goor) Vørs (basionym Codonomonas pascheri van Goor) es mejor considerarla como especie separada e incluirla en el genero Codonomonas van Goor. Desde un punto de vista ultraestructural la arquitectura básica de la lorica es la de una banda fibrilar que envuelve a la célula en espiral y que acaba endureciéndose debido a la mineralización inorgánica del medio. La aparición del componente fibrilar y el grado de mineralización puede variar considerablemente. El vértice celular se modifica en un citostoma apical lo que confirma una observación previa de que dichas células son capaces de ingerir bacterias. Aunque la correlación entre los recuentos de nanoflagelados determinados por microscopía de barrido (SEM) y los de bacterias por microscopía de epifluorescencia en el mejor de los casos son marginalmente significativos, es posible que las poblaciones de O. vangoorii de Barcelona subsistan parcialmente gracias a la bacterivoria y por tanto su papel ecológico es el de ser un consumidor eventual de carbono. Nosotros también hipotetizamos que la presencia de una lórica incrementa la posibilidad de interceptar las presas bacterianas, dispuestas para ser ingeridas por el citostoma apical de la célula.

Summary Performance of a full-scale wastewater treatment plant by rotating biological contactors (RBC) system was monitored during a year by physico-chemical and microbial characterisation. Six points along wastewater treatment were selected in the plant: three points along the water line (influent, sedimentation tank and effluent) and three points along RBC system (RBC1, RBC2 and RBC3). Although a large seasonal change in the values of physico-chemical parameters was observed, operation of the plant was optimal during all year (90% of removal in BOD5 and SS influent content). Microbial characterisation was approached by determining the structure and dynamics of protozoan and metazoan communities. Protozoa were the most abundant in all stages in the plant, heterotrophic flagellates being the most representative group in the water line and ciliates in the RBC system. The same seasonal preference was only observed for heterotrophic flagellates in the water line and green flagellates in the RBC system, both groups having highest abundances in summer and spring, respectively. Identification of ciliated protozoa populations rendered 58 species of ciliates in the plant. Most of these species are typical of aerobic wastewater treatment systems except three of them, which are cited for the first time in this type of ecosystems: Chaenea stricta, Holosticha mancoidea and Oxytricha lanceolata. Along the water line 34 species were identified, and half of them only appeared occasionally (once in all the study), while along the RBC system biofilms 55 species were observed, and the majority appeared permanently in this system. Our results indicate that the type of habitat, rather than the physico-chemical water parameters, was the primary factor in determining the different distribution of protozoan and metazoan communities in the plant. In RBC biofilms, the structure of ciliate protozoa community was found to be quite sensitive to changes in physico-chemical parameters, mainly to organic loading (BOD5) variations.

Ciliate communities were studied in the biofilms of three plants with rotating biological contactors (RBCs) over the period of one year. The aims of this study were: (i) to characterize ciliates to species level; (ii) to determine their abundance and spatial variation through the biological system; and (iii) to relate the spatial segregation and richness of ciliate species to plant performance. A number of species ranging from 33 to 67 were identified at the different plants. The overall best represented groups were peritrichs followed by scuticociliates (Las Matas), cyrtophorids (Boadilla) or hypotrichs (Camarma). Comparison of RBCs and activated sludge communities indicated that even though both were mainly constituted by peritrichs, differences in groups and species composition were observed.

Summary Stomatogenesis in Uronema filificum was studied with pyridinated silver carbonate. Fourteen morphogenetic stages are described. The ontogenetic sequence develops for the proter: haplokinety → haplokinety and scutica; for the opisthe: haplokinety → haplokinety, Ml, M2 and scutica, and scutica → M3. This stomatogenetic pattern is similar to that described for other genera in the same family, but the anlage morphogenetic sequence differs slightly from previous descriptions. The systematic position of Uronema filificum is discussed in the light of the new morphogenetic data reported.

The growth kinetics of two species of the genus Uronema, the marine U. marinum and the freshwater U. nigricans, have been studied under equivalent culture conditions. Ciliates were fed on three bacterial strains at three different concentrations (106, 107, and 108 bacteria ml−1). Growth rates in U. marinum were between 10 and 70% higher than those observed for the freshwater clone of U. nigricans, while net stationary phase populations were 1–60 times higher in U. marinum than in U. nigricans. The half-saturation constants (KS) for each bacterial strain showed that U. nigricans reached its $${\raise0.5ex\hbox{$\scriptstyle 1$}\kern-0.1em/\kern-0.15em\lower0.25ex\hbox{$\scriptstyle 2$}}$$ μmax at one to three times lower concentration than that observed for U. marinum. Despite the close morphological similarity of these species, U. marinum and U. nigricans exhibited sustained distinct growth behavior that might reflect the survival strategies followed in their specific environments.

Growth rates in 15 clones from the bacterivorous ciliate Uronema and one from Parauronema acutum have been compared under similar culture conditions. Five species were identified on a morphological basis: U. marinum, U. nigricans, U. elegans, P. acutum, and an undescribed form designated here as SWi1. Intra- and inter-specific distinct growth characteristics were observed. Net cellular production tended to decrease with lower growth rate in different species or morphotypes but increased in those clones belonging to the same species. Growth rate and cell volume were not significatly correlated although mean growth rate increased with mean cell volume. Growth rate was however significantly correlated to the relative variation in cell volume between different growth phases. Estimation of growth rates using different models indicated that the experimental growth rates observed were not significatly different from those expected for the cell volumes. These results indicate how effects of habitat or geographical variability, in this cosmopolitan genus, may account for trophodynamic variation of morphologically similar species.

The quality of the sludge in Wastewater Treatment Plants (WWTPs) depends on the suitable colonization of the flocs by microorganisms. Due to the functional importance of these biological constituents, several biological or biological-related parameters have been commonly used for the control of depuration efficiency. According to national and international water regulation recommendations, interlaboratory studies have a great relevance to determine which parameters are more reliable for their extensive application in routine control. However, these studies are also very useful to demonstrate consistency in results from multiple laboratories and to develop reliable and reproducible methodologies which might be necessary for protocol validation and also for accreditation issues to meet regulatory environmental requirements. The main purpose of this work was to assess the results obtained in consecutive interlaboratory assays in order to determine the concordance degree in the application of biological parameters by participating laboratories. Following the international recommendations about these studies, a common working protocol was proposed. Statistical tests indicated that Sludge Index and several routine physical–chemical analyses [V30, Mixed Liquor Suspended Solids (MLSS), Mixed Liquor Volatile Solids (MLVS) and Sludge Volumetric Index (SVI)] show low variability and therefore are suitable tools for laboratory control. Shannon Index and Sludge Biotic Index also presented low variability although a more precise protocol would be necessary, in particular the methodology to count small flagellates. The abundance and identification of protist species showed low concordance among laboratories and three factors were responsible for the low reliability of data: population density, size and morphological distinguishable characters of the specimens.

Unusual prostomatid specimens were found in the biological reactor of a wastewater treatment plant in a health resort in Valencia, Spain. These ciliates were attached to flocs unlike other free-swimming prostomatid ciliates described to date in the mixed liquor of activated sludge plants. The morphological study of this species led to a typically different combination of characteristics: elongated cell shape, 20-30 somatic kineties, 2 perioral kineties, and 1 circumoral kinety, 1 large vacuole protruding at the terminal end, a lorica tapered toward the aperture with a smooth neck, and 11-16 annular ridges. These characteristics place this representative as a new species of the genus Metacystis-Metacystis galiani n. sp. This species became the dominant population within the biological reactor when high values of conductivity (4,244 mS/cm) and temperature (26.8 degrees C) were recorded.

Three full-scale advanced biological systems for nitrogen removal showing different efficiencies were assessed during one year, to investigate the protist communities supported in these wastewater treatment plants (WWTP). The main goal of this research was to explore the differences of these communities from those observed in conventional activated sludge systems. The final objective was to provide background support for the proposal of bioindicators in this type of biological systems, where scarce information was available until now, since only conventional systems had been previously studied from this point of view. Results obtained indicate that, in fact, protist population density and diversity in advanced systems for N-elimination are quite different from other wastewater systems studied before. A statistical approach through multivariate analysis was developed to search for association between protist species and physical-chemical system performance, and specifically N-removal efficiencies. The original hypothesis proposing that previous indicators from conventional systems are not adequate in advanced N-removal mechanisms was proved to be correct. Efficient processes on N-removal, despite what it had been usually found in conventional systems, show important flagellate and amoeba populations and these populations tend to reduce their abundances as nitrogen removal performance decreases (moderate to low). Ciliates are however less abundant in these N-removal efficient systems. Certain groups and genera of protist such as flagellates and small amoebae are thus proposed as indicative of high performance N-removal, while in this case the appearance of certain ciliates were indicative of low performance on N- or high organic matter removal (as COD) efficiencies.

The biological community in activated sludge wastewater plants is organized within this ecosystem as bioaggregates or flocs, in which the biotic component is embedded in a complex matrix comprised of extracellular polymeric substances mainly of microbial origin. The aim of this work is to study the role of different floc-associated ciliates commonly reported in wastewater treatment plants-crawling Euplotes and sessile Vorticella- in the formation of aggregates. Flocs, in experiments with ciliates and latex beads, showed more compactation and cohesion among particles than those in the absence of ciliates. Ciliates have been shown to contribute to floc formation through different mechanisms such as the active secretion of polymeric substances (extrusomes), their biological activities (movement and feeding strategies), or the cysts formation capacity of some species. Staining with lectins coupled to fluorescein showed that carbohydrate of the matrix contained glucose, manose, N-acetyl-glucosamine and galactose. Protein fraction revealed over the latex beads surfaces could probably be of bacterial origin, but nucleic acids represented an important fraction of the extracellular polymeric substances of ciliate origin.

Protists, mainly ciliates, play several essential roles in biological wastewater treatment, such as the transfer of matter and energy, bacterial predation, and the removal of organic material. Moreover, during the treatment process, the formation of bioaggregates-flocs and biofilms-is essential to obtaining high-quality effluents. In the present study, Tetrahymena thermophila was used as a model organism to demonstrate the contribution of ciliates to bioflocculation. Axenic cultures of this species were exposed to chemical and mechanical stimuli that promote bioaggregation. In either case, the secretion of a capsulate mucous material by the ciliates or by particle aggregation was detected. Numerous, small, loosely compacted flocs were observed under shaking conditions and in the presence of latex beads. The composition of the exopolymeric material secreted by ciliates was analyzed by a series of fluorochromes and colorimetric methods, which showed that carbohydrates and nucleic acids were the main components involved in matrix formation and particle adhesion.

A rotating biological contactor (RBC) system operating in a full-scale wastewater treatment plant has been described by several approaches accounting for performance, composition and structure of biofilms in three stages through biological wastewater treatment (RBC1, RBC 2, RBC 3). RBC biofilms were effective in removing the BOD loading from 13 g BOD 5 d −1 m −2 in RBC 1 to 6 g BOD 5 d −1 m −2 in RBC 3. Analysis of biofilm composition showed: i) the volatile solids were similar in the three RBCs (0.6 g m −2 VS per g m −2 of TS); ii) the protozoan and metazoan biocenosis was mainly made up of ciliated protozoa, which were most abundant in RBC 2 (1.84 × 10 6 ciliates g −1 VS). Relationship between ciliate species and physical-chemical profile of the system by cluster analysis indicated that the species Acineria uncinata , Amphileptus punctatus , Cinetochilum margaritaceum and Holosticha mancoidea were associated with the best RBC performance; iii) the exopolymeric matrix of the three RBC biofilms was mainly constituted by proteins, although humic substances, polysaccharides, uronic acids and DNA were also found. Analysis of biofilm structure by confocal microscopy indicated changes in biofilm organisation with depth. Results have been brought together and a graphic representation of the composition and architecture of RBC biofilms is presented.

A new genus and species combination are proposed for Urocryptum tortum n. gen., n. comb., a scuticociliate with a polymorphic life cycle. This marine ciliate was isolated from a sample taken at Gokasho Bay in Mie Prefecture (Japan). Specimens from different phases of the growth cycle were examined in vivo and with two silver staining techniques. Three life-history stages were observed: an exponential growth phase stage (trophont), a stationary phase stage (tomite), and finally a resting stage (cyst). The exponential growth form is laterally flattened and ovoid; it has 20-24 somatic kineties (SK) and a typical complement of scuticociliate oral structures. Polykinetid 1 (Pk1) has two longitudinal files of 6 kinetosomes (Ks); sometimes one or two additional kinetosomes are located anteriorly. Polykinetid 2 (Pk2) has two files of 6 or 7 Ks; a third file of three widely spaced kinetosomes is located on its right side, as well as a small curved row of 5 Ks positioned on the right hand side of the posterior end of Pk2. Polykinetid 3 (Pk3) has 3 rows of 4, 5, and 7 Ks's perpendicular to the haplokinety (Hk) or paroral membrane. The stationary growth phase cell is spindle-shaped and has a similar number of SK that are much more closely spaced than in exponentially growing specimens. Oral infraciliary structures are reduced in size, having fewer kinetosomes and being positioned more anteriorly in the cell. Pk1 is composed of two files of 5-Ks, Pk2 has only two files of 6-7 Ks, and Pk3 has two to three rows of 3-4 Ks. The Hk is displaced anteriorly and becomes straight, losing the shape typical of exponential growth phase cells. Cysts are pyriform with a wide cyst wall; no infraciliary structures were visible.

Summary Physical-chemical monitoring and characterization of ciliate communities from a full-scale rotating biological system (RBC) have been carried out for a year. RBC system operated efficiently in removing the organic matter, as the decrease of the BOD 5 loading along the successive RBC units revealed. 55 species of ciliated protozoa were identified in the RBC biofilms. Differences in abundance, occurrence and type of species were found along the different units of the RBC system; the last RBC held a more stable and diverse ciliate community. The complexity of interspecific relationships among the ciliates has been outlined using multivariate methods (Cluster and Correspondence analysis). Correlation between ciliate species and physical-chemical conditions were obtained by regression analysis. Results show that 12 species of ciliates were related to an optimal efficiency in organic matter removal, Litonotus crystallinus being the most sensitive species. The presence and abundance of Litonotus crystallinus and Acineria uncinata was associated with a decrease in the organic factor. Metopus es was the only species related to a decrease in process efficiency due to its association with an increase in organic factor. The results provide statistical evidence of the use of certain ciliate species as reliable bioindicators in full-scale RBC wastewater treatment plants.