Diversity and geographic distribution of ciliates (Protista: Ciliophora)

About 4,500 free-living ciliate morphospecies have been described, applying an average synonymy rate of 20%. We estimate that 83–89% of the ciliate diversity is still undescribed, using the following probabilities: detailed habitat studies suggest that the described number of morphospecies must be doubled: 4,500 → 9,000; this figure has to be increased by about 50% due to species with similar interphase morphology but different resting cysts: 9,000 → 13,500; the genetic and molecular data suggest that this value must be doubled or trebled: 13,500 → 27,000 to 40,000 free-living, biological ciliate species. The knowledge on geographic distribution of ciliates heavily depends on flagship species and statistical analyses because reliable faunistic studies are rare and molecular data are still in its infancy. We present a list of 52 ciliate flagship species as a testable hypothesis, i.e., the hypothesis of restricted distribution of certain ciliate species must be refused when a considerable number of them is found in all or most biogeographic regions. Flagship species and statistical analyses consistently show Gondwanan and Laurasian ciliate communities, suggesting that the split of Pangaea deeply influenced ciliate distribution and rare species play a key role in geographic differentiation. However, there is also substantial evidence for continental, regional, and local endemism of free-living ciliates. The molecular studies usually show a high level of genetic diversity underlying ciliate morphospecies, suggesting that morphologic and molecular evolution may be decoupled in many ciliate species. Molecular studies on ciliate biogeography are at variance, possibly because most are still focusing on single molecular markers. In sum, the data indicate that ciliate biogeography is similar to that of plants and animals, but with an increased proportion of cosmopolites, favouring the moderate endemicity model. Special Issue: Protist diversity and geographic distribution. Guest editor: W. Foissner.     

Spatial variation in ciliate communities with respect to water quality in the Delhi NCR stretch of River Yamuna,India

The River Yamuna emerges from Saptarishi Kund, Yamunotri and merge with River Ganges at Allahabad, India. Anthropogenic stress has affected the water quality of the river Yamuna drastically in the stretch traversing Delhi and its satellite towns (National Capital Region, NCR). In the present study, effect of water quality on the microbial life in the River Yamuna was analyzed using ciliate communities (Protista, Ciliophora) as bio-indicators. Water samples were collected from six sampling sites chosen according to the levels of pollution along the river and water quality was analysed using standard physicochemical factors. As the river traverses Delhi NCR, water quality deteriorates considerably as indicated by the Water Quality Index at the selected sampling sites. Seventy-four ciliate species representing nine classes were recorded. Based on the Shannon diversity index, maximum species diversity was found at the point where the river enters Delhi. The saprobity index showed the river water was beta-mesosaprobic when the river enters Delhi and alpha-mesosaprobic at downstream sites after the first major drain outfall. Significant relationship between the spatial variation in ciliate communities and abiotic parameters indicate that ciliates can be used as effective bioindicators of pollution in the River Yamuna.     

Analysis and evolution of water quality of the upper Váh River (northern Slovakia) by long-term changes in the community structure of ciliates (Protista: Ciliophora)

A long-term monitoring of the upper Váh River was carried out in order to evaluate evolution of water quality by means of ciliate taxocoenoses. One sampling station (Lisková) was located upstream of the pulp and paper mill in the town of Ru?omberok, while two stations (Bystrá and Stankovany) were situated downstream of Ru?omberok. Benthic samples were collected three times a year at each station during 2004–2011. This long-term study enabled us to assemble a comprehensive checklist of ciliates recorded in the upper Váh, and to characterize and compare the ciliate communities in the investigated watercourse. The saprobic index and valency analysis of the ciliate communities allowed us to quantify organic input and to evaluate changes in the saprobity of the upper Váh. The saprobic index ranged from 2.45 to 3.66 corresponding to the quality classes II–IV. Further, there was a prevalence of the α- and β-mesosaprobic levels, but during some periods the polysaprobic level increased along with the decrease of the β-mesosaprobic level. Beside the organic pollution, toxic contaminations were also noticed several times in the studied watercourse. This was documented by a significant drop in the species diversity and abundance of ciliates as well as by various morphological malformations and movement disorders of ciliates and other microorganisms. However, the negative effect of toxic and/or organic pollution wore off relatively quickly, indicating a good self-purification capability of the river.     

Experimental infections of Orchitophrya stellarum (Scuticociliata) in American blue crabs (Callinectes sapidus) and fiddler crabs (Uca minax)

Outbreaks of an unidentified ciliate have occurred on several occasions in blue crabs from Chesapeake Bay held during winter months in flow-through systems. The parasite was initially thought to be Mesanophrys chesapeakensis, but molecular analysis identified it as Orchitophyra stellarum, a facultative parasite of sea stars (Asteroidea). We investigated the host-parasite association of O. stellarum in the blue crab host. Crabs were inoculated with the ciliate, or they were held in bath exposures after experimentally induced autotomy of limbs in order to determine potential mechanisms for infection. Crabs inoculated with the ciliate, or exposed to it after experimental autotomy, rapidly developed fatal infections. Crabs that were not experimentally injured, but were exposed to the ciliate, rarely developed infections; thus, indicating that the parasite requires a wound or break in the cuticle as a portal of entry. For comparative purposes, fiddler crabs, Uca minax, were inoculated with the ciliate in a dose-titration experiment. Low doses of the ciliate (10 per crab) were sometimes able to establish infections, but high intensity infections developed quickly at doses over 500 ciliates per crab. Chemotaxis studies were initiated to determine if the ciliate preferentially selected blue crab serum (BCS) over other nutrient sources. Cultures grown on medium with BCS or fetal bovine serum showed some conditioning in their selection for different media, but the outcome in choice experiments indicated that the ciliate was attracted to BCS and not seawater. Our findings indicate that O. stellarum is a facultative parasite of blue crabs. It can cause infections in exposed crabs at 10–15 °C, but it requires a portal of entry for successful host invasion, and it may find injured hosts using chemotaxis.     

A statistical approach to estimate soil ciliate diversity and distribution based on data from five continents

A total of 359 soil samples collected from five continents (Africa, Asia, Australia, Europe and South America) were investigated for the presence/absence of soil ciliate species. Merging records by species identity, we have compiled a master data list (species by sample matrix). In the list, a total of 964 soil ciliate species (644 described and 320 undescribed) are recorded. The species distributions within the 359 samples and across the five continents are examined. The frequency distribution of the species over samples is used for global diversity estimation. A statistical ACE (abundance‐based coverage estimation) model which links observed data to unseen species is applied to assess regional and global soil ciliate species richness. The model, whose reliability was tested by its power to predict the number of new species in additional samples from Africa, may resolve the controversial issue on global species diversity of soil ciliates. Although an accurate point estimate is not feasible due to severe undersampling, the statistical model enables us to obtain a minimum regional diversity and global species diversity. A consistent finding over all five continents is that at least half of the species diversity is still undiscovered. Our model also yields a global soil ciliate diversity of at least 1900 species, that is, doubles the number of currently known species, and thus diversity is relatively high. This is consistent with the finding of Foissner, who used a probability‐based method. Soil ciliate distributions between continent pairs are analyzed by adjusted abundance‐based similarity/overlap indices. These new indices account for the effect of unseen species and also reduce the bias generated by undersampling. The adjusted abundance‐based Jaccard (or Sørensen) index shows that there is about 30% (18% for Sørensen) dis‐similarity between any two continents, supporting the moderate endemicity model. The results are discussed with respect to protist species distribution, that is, whether they are cosmopolitan or of restricted distribution.     

Genetic analysis of ciliates living on the groundwater amphipod Crangonyx islandicus (Amphipoda: Crangonyctidae)

Two endemic subterranean freshwater amphipod species have been discovered in groundwater of the volcanic active zone in Iceland, Crangonyx islandicus and Crymostygius thingvallensis. At least five different ciliate sequences were isolated from C. islandicus and the analysis of variation at the 18S ribosomal DNA gene suggests that they present previously unsequenced species, from the orders Apostomatida and Philasterida. Apostome ciliates are well known to be exuviotrophic epibionts on crustaceans. Analyses of ciliate epibionts from different groundwater amphipod species from North America and mainland Europe revealed distinct groups of ciliates and higher diversity on the epigean amphipod Crangonyx pseudogracilis. Analysis of geographic patterns of the most common ciliate within Iceland revealed population differentiation supporting limited current connectivity between the different groundwater systems. Our study reports an occurrence of previously undescribed ciliate species in a groundwater ecosystem characterized by low species diversity.     

The influence of environmental characteristics on the distribution of ciliates (Protozoa, Ciliophora) in an urban stream of southeast Brazil

The aim of this research was to study the ciliated protozoa community at three sampling stations that receive different levels of domestic sewage along the S?o Pedro Stream in the municipality of Juiz de Fora, Minas Gerais, Brazil, in order to determine the influence of organic pollution on this community and to assess the feasibility of using ciliates as water quality indicators. Four physical-chemical parameters of the water samples were evaluated: dissolved oxygen concentration, electrical conductivity, pH and temperature. The sediment was obtained manually, using dredges with capacity of 300 mL, at each collection point. Point 1 was located in a rural region that receives a low sewage load, while Points 2 and 3 were located in populated regions receiving high sewage loads. We found 22 ciliate species, of which 18 are included in the saprobic system and are considered bioindicators. These showed beta-mesosaprobic environments at Point 1 and alfa-mesosaprobic to polisaprobic environments at Points 2 and 3. The low levels of dissolved oxygen and the high electrical conductivity values at Points 2 and 3, together with the strong similarity between the ciliate taxocenoses of these points and the weak similarity between Point 1 and the other two, confirm the high sewage loads received at the latter two points. The combination of the biological indicators and physical-chemical analyses therefore proved itself to be an efficient method of evaluating water quality, and has excellent potential to support decisions on the conservation of headwaters and recuperation of degraded environments in lotic systems.     

Distribution and summer diapause of Canthocamptus staphylinus (Jurine) (Copepoda: Harpacticoida) in Lake Ladoga

The spatial distribution of Canthocamptus staphylinus (Jurine) in Lake Ladoga was examined. C. staphylinus is one of the most numerous species of harpacticoids in the lake. It can dwell in almost all littoral biotopes. In profundal biotopes strongly exposed to anthropogenic influence (pollution and eutrophication) only this species of harpacticoid was found. A peculiarity of C. staphylinus biology is that in summer individuals are in the resting stage as cysts. In summer active individuals were found in littoral habitats at a water temperature of 20 °C, but their number was small in comparison with cold periods of the year. In the littoral zone the relative number of individuals in cysts was clearly related to water temperature. Twelve degrees temperature appears to be critical for development of littoral populations of C. staphylinus.     

Development of an Angiosperm Quality Assessment Index (AQuA-Index) for ecological quality evaluation of Portuguese water bodies—A multi-metric approach

Salt marsh vegetation is one important biological element for establishing the ecological quality status (EQS) while evaluating transitional waters (TW). This intertidal plant community is sensitive to the most important pressures present in estuaries. In this study, an ecological index based multi-metric was established taking into account the species composition and ecological relations in Portuguese salt marsh habitats. During the AQuA-Index elaboration it was found that the parameters that respond better to the variability of ecological conditions present nowadays are the Shannon Diversity Index, the Shannon Maximum Diversity Index, the number of halophytic species, the Margalef Diversity Index and the Pielou Equitability Index. The application of the AQuA-Index index to eight TW systems varying in morphology and pressures, demonstrated that the metric responds to the ecological quality of each system. Also, it showed some advantages in comparison with other approaches, in zones with a relevant proportion of middle and upper marsh and where historical data were not available. This way the AQuA-Index proved to be an efficient Index for ecological quality assessment, using salt marsh vegetation as indicator.     

Ciliate communities of a large shallow lake: Association with macrophyte beds

We investigated the influence of macrophyte composition on ciliate community structure in a large, shallow, eutrophic Lake Võrtsjärv. We hypothesized that macrophyte composition must have strong influence on the dispersal of ecologically different ciliate groups in a shallow lake and that more diverse macrophyte stands cause also a greater diversity in the ciliate community. In Võrtsjärv macrophyte distribution is spatially strongly polarized both in east–west and north–south directions in relation to abiotic factors. Phragmites australis and Myriophyllum spicatum were the most widespread species occurring in most parts of the lake. Correlation of environmental, macrophyte and planktonic ciliate variables confirmed the suggested spatial gradients. More diverse macrophyte stands supported a high species richness and abundance of epiplanktonic community but showed negative influence on the number and abundance of euplanktonic ciliate taxa. Opposite trends were found relative to the abundance of P. australis. Benthic ciliates showed a similar distribution pattern to euplanktonic taxa being most abundant in sites were the Shannon–Weaver index for macrophytes was low. Strong polarizing effect of the lake's vegetation on planktonic ciliate diversity was reflected in correlations of the number of ciliate taxa as well as the numbers of eu- and epiplanktonic taxa with geographic co-ordinates.     

Comparative analysis of the mitochondrial cytochrome c oxidase subunit I (COI) gene in ciliates (Alveolata,Ciliophora) and evaluation of its suitability as a biodiversity marker

The mitochondrial cytochrome c oxidase subunit 1 (COI) gene of ciliates was first successfully sequenced in species of the genera Tetrahymena and Paramecium (Class Oligohymenophorea). The sequence of the COI gene is extremely divergent from other eukaryotes and includes an insert, which is over 300 nucleotides long. In this study, we designed a primer pair that successfully amplified the COI gene of ciliates from five different classes: Heterotrichea, Spirotrichea, Oligohymenophorea, Nassophorea and Colpodea. These classes represent the diversity of the phylum Ciliophora very well, since they are widely distributed on the ciliate small subunit rRNA tree. The amplified region is approximately 850 nucleotides long and corresponds to the general barcoding region; it also includes the insert region. In this study, 58 new COI sequences from over 38 species in 13 orders are analysed and compared, and distance trees are constructed. While the COI gene shows high divergence within ciliates, the insert region, which is present in all classes, is even more divergent. Genetic distances calculated with and without the insert region remain in the same range at the intraspecific level, but they differ considerably at or above genus level. This suggests that the entire barcoding region is under similar selective constraints and that the evolutionary rate of the ciliate COI is extremely high and shows unequal rate variation. Although many problems still remain regarding standardization of barcoding methods in ciliates, the development of a universal or almost universal primer combination for the Phylum Ciliophora represents important progress. As shown in four examples, the resolution of COI at the intraspecific level is much greater than that of any nuclear genes and shows great potential to (1) identify species based on molecular data if a reliable database exists, and (2) resolve the relationships of closely related ciliate taxa and uncover cryptic species.     

土壤纤毛虫群落对退耕还林生态恢复的响应——以侧柏林为例

为了了解土壤纤毛虫群落对退耕还林区生态恢复下土壤环境变化的响应及尝试利用土壤纤毛虫群落特征评价退耕还林生态恢复的效果,于2010年3月至9月采用"非淹没培养皿法"、活体观察法、蛋白银染色法和Foissner计数法对甘肃陇南武都退耕还林区5个不同恢复阶段的样点和1个荒草坡对照样点的土壤纤毛虫群落特征进行了研究,同时测定了p H值、温度、含水量及速效磷、有效钾、铵态氮、有机质和有机碳含量等土壤环境因子并分析了生态恢复条件下土壤纤毛虫群落特征与土壤环境因子的相关性。研究中共鉴定到71种纤毛虫,隶属于3纲、10目、22科、29属。研究发现,不同恢复年限的土样中土壤纤毛虫的物种分布存在明显差异,并随恢复时间的延长,样点间物种相似性减小,群落组成复杂化,物种数、种群密度和物种多样性指数总体均呈增长趋势。优势类群也发生了演替,由恢复初期的肾形目演替到后期的下毛目。相关性分析结果表明,在生态恢复条件下,土壤有机质和铵态氮含量是影响土壤纤毛虫群落结构稳定性的主要因素,不同纤毛虫类群对生态恢复的响应存在差异。冗余分析显示,土壤纤毛虫群落很好地响应了生态恢复过程中土壤环境条件的变化。     

Biological evaluation of running waters in Korea, with special reference to bioassessment using the PFU system

A total of six stations in the Han River system were selected for establishing polyurethane foam units (PFUs) to collect protozoans, including phytomastigophorans, zoomastigophorans, amoebas and ciliates, in July 1993. In the bioassessment of microbial communities using the PFUs, the number of species decreased as pollution intensity increased. The diversity index values calculated at the main stations generally agreed with the pollution status of the stations. Anyang-Chon (Chon means stream) showed the lowest diversity value (1.89), and all stations, except Masok and Anyang-Chon, showed diversity index values ranging from 3.15 to 3.93. The highest heterotrophic index (HI) value was detected in Anyang-Chon followed by Masok-Chon. The number of species at the main stations reached a maximum on day 11 of being exposed to PFUs. The results of _eq, G and T_90% all suggest that bioassessments using the PFU system were well matched with pollution status of the water. All microbial variables were significantly correlated with comprehensive chemical pollution indices, P_a and P_b , with correlation coefficients ranging from r =0.87 to r=0.96. This revised version was published online in July 2006 with corrections to the Cover Date.     

Rumen ciliated protozoan fauna of the yak (Bos grunniens) in China with the description of Entodinium monuo n. sp

Rumen ciliate species composition was surveyed in domestic yaks kept in Tibet, Sichuan, and Inner Mongolia, China. Twelve genera including 36 species with 18 formae were identified. The species compositions were slightly different among the three areas: yaks in Tibet had the simplest fauna, in contrast, the fauna of yaks in Inner Mongolia were the most abundant and similar to those found in the cattle kept in the same area. This suggests that the rumen ciliate composition of yaks is affected by that of cattle kept together or in proximity. A new species belonging to the genus Entodinium, Entodinium monuo n. sp., was recognized from the yaks in all areas. This new species was common in the yaks but was not detected in the cattle fed near yaks in Inner Mongolia. There was a similar generic ciliate composition in yaks kept in respective areas. Entodinium was the most predominate ciliate (51.9-61.0%) with total ciliate densities estimated as 10(5)/ml per yak.     

Diversity and Distributional Patterns of Ciliates in Guaymas Basin Hydrothermal Vent Sediments

Little is known about protists at deep‐sea hydrothermal vents. The vent sites at Guaymas Basin in the Gulf of California are characterized by dense mats of filamentous pigmented or nonpigmented Beggiatoa that serve as markers of subsurface thermochemical gradients. We constructed 18S rRNA libraries to investigate ciliate assemblages in Beggiatoa mats and from bare sediments at the Guaymas vent site. Results indicated a high diversity of ciliates, with 156 operational taxonomic units identified in 548 sequences. Comparison between mat environments demonstrated that ciliate and bacterial assemblages from pigmented mats, nonpigmented mats, and bare sediments were significantly different and highly correlated with bacterial assemblages. Neither bacterial nor ciliate assemblages were correlated with environmental factors. The most abundant ciliates at Guaymas were more likely to be represented in clone libraries from other hydrothermal, deep‐sea, and/or anoxic or microaerophilic environments, supporting the hypothesis that these ciliate species are broadly distributed. The orange mat environment included a higher proportion of ciliate sequences that were more similar to those from other environmental studies than to cultured ciliate species, whereas clone libraries from bare sediments included sequences that were the most highly divergent from all other sequences and may represent species that are endemic to Guaymas.     

Contribution of NaCl excretion to salt resistance of Aeluropus littoralis (Willd) Parl

Aeluropus littoralis is a perennial halophyte, native to coastal zones. Although it is usually exposed to high saline, this plant grows normally without toxicity symptoms. In order to assess leaf salt excretion, different growth parameters, Na(+), K(+), Ca(2+), Mg(2+) and Cl(-) concentrations, as well as excreted ions were examined in plants grown for 2 months in the presence of various salinity levels (0-800 mM NaCl). In addition, salt crystals, salt glands and other leaf epidermal structures were investigated. Results showed that total plant growth decreased linearly with increase to medium salinity. This reduction concerns mainly shoot growth. In addition, this species was able to maintain its shoot water content at nearly 50% of the control even when subjected to 800 mM NaCl. Root water content seemed to be unaffected by salt. Sodium and chloride ion contents in shoots and in roots increased with salinity concentrations, in contrast to our observation for potassium. However, calcium and magnesium contents were not greatly affected by salinity. Excreted salts in A. littoralis leaves were in favor of sodium and chloride, but against potassium, calcium and magnesium which were retained in plants. Sodium and chloride were excreted from special salt glands, which were scattered on the both leaf surfaces. In addition to salt glands, papillae were the most frequent epidermal structure found on A. littoralis leaves, and are likely involved in A. littoralis salt resistance.     

The effect of salinity on different developmental stages of an endemic annual plant, Aster laurentianus (Asteraceae)

Salinity reduces substrate water potential, thereby restricting water and nutrient uptake by plants; salinity may also cause ionic imbalance and toxicity. Because substrate salinity fluctuates through the growing season, a plant may be exposed to different salinity levels, at various stages of development, with potentially significant consequences on population dynamics. Here, we present the results of a study of the effect of substrate salinity on seed germination, seedling emergence, and growth of Aster laurentianus, an annual marsh plant, endemic to the Gulf of St. Lawrence and potentially threatened. Seed germination was reduced in low salt concentration (10 g sea salt/L) and completely inhibited by salinity levels >/=20 g sea salt/L. However, this inhibiting effect was reversible: seeds from the salt treatments germinated readily after being washed in distilled water. Though seedling emergence was diminished at low salinity levels, postemergence survival was little affected. Plant growth was reduced, but net carbon assimilation rate was not affected by high salinity levels. Increased root respiration and respiratory costs associated with salt tolerance might have contributed to lower C accumulation at higher salinity levels. All developmental processes considered are thus negatively affected by substrate salinity, with potentially significant consequences on population abundance and distribution in salt marshes. Yet, the tolerance of this species to high salinity levels after seedling emergence is remarkable. Seed germination represents a major bottleneck in the species life cycle, potentially controlling local distribution and abundance in the natural habitat.     

Salinity tolerance in Hyalophysa chattoni (Ciliophora, Apostomatida), a symbiont of the estuarine grass shrimp Palaemonetes pugio

The apostome ciliate Hyalophysa chattoni, a symbiont of the estuarine grass shrimp Palaemonetes pugio, was tested for its growth and reproductive ability in a wide range of salinities from 0.1 to 55 ppt. Shrimp, with their attached ciliates, were slowly acclimated to different salinities in order to assess protozoan cell size and division. The trophont and tomont stages of the ciliate life cycle were analyzed. In both stages, cell size increased with salinity from 0.1 to 20 ppt. Cell size leveled in the 20-35 ppt range, and decreased at higher salinities. The number of daughter cells produced per tomont cyst correlated with increased cell size, and also correlated with increased salinity. Additionally, increased salinity correlated with an increase in the percentage of cells able to divide and excyst as tomite stages. These results indicate that H. chattoni is able to grow and divide more effectively at salinities closer to seawater than in the estuarine environment from which they were collected. Though able to survive salinities from 0.1 to 55 ppt, the species is better adapted for an existence in the higher salt concentrations.     

Spider biodiversity potential of an ungrazed and a grazed inland salt meadow in the National Park 'Neusiedler See-Seewinkel' (Austria): implications for management (Arachnida: Araneae)

To assess the biodiversity potential of an ungrazed and a grazed inland salt meadow in the Seewinkel (Eastern Austria), spider assemblages were recorded by pitfall trapping for 1 year. Both species assemblages consisted, to a large extent, of rare species of conservation interest. The species assemblage of the grazed site was dominated by Pardosa agrestis, but highly specific halotopobiontic species also occurred in higher numbers. Halotolerant species were also present in the ungrazed meadow, but their individual number was much lower. The species composition of this site reflects the more balanced microclimatical situation of the high sward. Comparison of the two assemblages with 207 other meadow spider assemblages of Central Europe shows a separated position, especially of the grazed site assemblage. High similarities with assemblages of meadows with a similar vegetation structure indicate a high importance of management. Considering the high proportion of rare species on both sites, the best management of the salt meadow and pan shores of the Seewinkel should combine areas of light grazing with ungrazed areas. However, the proportion of these parts and the intensity of grazing still remains to be determined by quantitative experiments.