Brandtia ciliaticola gen. et sp. nov. (Chlorellaceae,Trebouxiophyceae) a common symbiotic green coccoid of various ciliate species

Many freshwater protists harbor unicellular green algae within their cells and these host‐symbiont relationships slowly are becoming better understood. Recently, we reported that several ciliate species shared a single species of symbiotic algae. Nonetheless, the algae from different host ciliates were each distinguishable by their different genotypes, and these host‐algal genotype combinations remained unchanged throughout a 15‐month period of sampling from natural populations. The same algal species had been reported as the shared symbiont of several ciliates from a remote lake. Consequently, this alga appears to play a key role in ciliate‐algae symbioses. In the present study, we successfully isolated the algae from ciliate cells and established unialgal cultures. This species is herein named Brandtia ciliaticola gen. et sp. nov. and has typical ‘Chlorella‐like’ morphology, being a spherical autosporic coccoid with a single chloroplast containing a pyrenoid. The alga belongs to the Chlorella‐clade in Chlorellaceae (Trebouxiophyceae), but it is not strongly connected to any of the other genera in this group. In addition to this phylogenetic distinctiveness, a unique compensatory base change in the SSU rRNA gene is decisive in distinguishing this genus. Sequences of SSU‐ITS (internal transcribed spacer) rDNA for each isolate were compared to those obtained previously from the same host ciliate. Consistent algal genotypes were recovered from each host, which strongly suggests that B. ciliaticola has established a persistent symbiosis in each ciliate species.     

The Ability of Tetrahymena utriculariae (Ciliophora,Oligohymenophorea) to Colonize Traps of Different Species of Aquatic Carnivorous Utricularia

The host specificity of the recently described ciliate species Tetrahymena utriculariae was tested in a greenhouse growth experiment, which included 14 different species of aquatic Utricularia as potential host plants. We confirmed the high specificity of the interaction between U. reflexa and T. utriculariae, the former being the only tested host species able to maintain colonization for prolonged time periods. We conclude that this plant–microbe relationship is a unique and specialized form of digestive mutualism and the plant–microbe unit a suitable experimental system for future ecophysiological studies.     

Morphology and Small Subunit rDNA Phylogeny of Two New Marine Urostylid Ciliates,Caudiholosticha marina sp. nov. and Nothoholosticha flava sp. nov. (Ciliophora,Hypotrichia)

Two marine urostylid ciliates, Caudiholosticha marina sp. nov. and Nothoholosticha flava sp. nov., isolated from intertidal sediment in the Yellow Sea, are investigated using morphological and small subunit rDNA phylogenetic analyses. Caudiholosticha marina is 210?310 μm × 40?55 μm in vivo, and has 10?20 macronuclear nodules, 23?37 midventral cirral pairs extending to 5?8 transverse cirri, and two caudal cirri. It differs from congeners by its marine habitat, larger size, macronuclear arrangement pattern and high number of midventral pairs. Molecular phylogenetic analyses indicate a polyphyly of Caudiholosticha. Nothoholosticha flava is yellow to brownish and 240?320 μm × 40?60 μm sized, and has a bipartite adoral zone, six frontal cirri in atypical bicorona, usually four frontoterminal, one buccal and 5?7 transverse cirri and 28?54 midventral pairs. Phylogenetic analyses allocate N. flava as sister of N. fasciola, type of the genus. The two Nothoholosticha species differ distinctly by the presence/absence of frontoterminal cirri, a feature often used to define genera in the Hypotrichia. However, the SSU rDNA sequence similarity between these two species is 99.3%, which weakens the justification for separating the new isolate at genus level. The taxonomic significance of frontoterminal cirri is discussed based on morphological and molecular data.     

Ecological Traits of the Algae‐Bearing Tetrahymena utriculariae (Ciliophora) from Traps of the Aquatic Carnivorous Plant Utricularia reflexa

Trap fluid of aquatic carnivorous plants of the genus Utricularia hosts specific microbiomes consisting of commensal pro‐ and eukaryotes of largely unknown ecology. We examined the characteristics and dynamics of bacteria and the three dominant eukaryotes, i.e. the algae‐bearing ciliate Tetrahymena utriculariae (Ciliophora), a green flagellate Euglena agilis (Euglenophyta), and the alga Scenedesmus alternans (Chlorophyta), associated with the traps of Utricularia reflexa. Our study focused on ecological traits and life strategies of the highly abundant ciliate whose biomass by far exceeds that of other eukaryotes and bacteria independent of the trap age. The ciliate was the only bacterivore in the traps, driving rapid turnover of bacterial standing stock. However, given the large size of the ciliate and the cell‐specific uptake rates of bacteria we estimated that bacterivory alone would likely be insufficient to support its apparent rapid growth in traps. We suggest that mixotrophy based on algal symbionts contributes significantly to the diet and survival strategy of the ciliate in the extreme (anaerobic, low pH) trap‐fluid environment. We propose a revised concept of major microbial interactions in the trap fluid where ciliate bacterivory plays a central role in regeneration of nutrients bound in rapidly growing bacterial biomass.     

Sources of mycosporine-like amino acids in planktonic Chlorella-bearing ciliates (Ciliophora)

1. Mycosporine‐like amino acids (MAAs) are a family of secondary metabolites known to protect organisms exposed to solar UV radiation. We tested their distribution among several planktonic ciliates bearing Chlorella isolated from an oligo‐mesotrophic lake in Tyrol, Austria. In order to test the origin of these compounds, the MAAs were assessed by high performance liquid chromatography in both the ciliates and their symbiotic algae. 2. Considering all Chlorella‐bearing ciliates, we found: (i) seven different MAAs (mycosporine‐glycine, palythine, asterina‐330, shinorine, porphyra‐334, usujirene, palythene); (ii) one to several MAAs per species and (iii) qualitative and quantitative seasonal changes in the MAAs (e.g. in Pelagodileptus trachelioides). In all species tested, concentrations of MAAs were always <1% of ciliate dry weight. 3. Several MAAs were also identified in the Chlorella isolated from the ciliates, thus providing initial evidence for their symbiotic origin. In Uroleptus sp., however, we found evidence for a dietary source of MAAs. 4. Our results suggest that accumulation of MAAs in Chlorella‐bearing ciliates represents an additional benefit of this symbiosis and an adaptation for survival in sunlit, UV‐exposed waters.     

Rumen protozoa of the giraffe with a description of two new species

An analysis was carried out on the rumen ciliate population of giraffe. Eight giraffe shot on the Timbavati and adjoining private nature reserves in the north eastern Transvaal, South Africa, were sampled during 1973. Twenty-six species of ciliates were found in the giraffe rumen. Two new species were found and they are described viz. Entodinium longicorpum sp.n. and Eodinium bispinosum sp.n. The genus Ento-dinium with eleven species formed 41% of the total protozoa population. The holotrichs formed 19%, the genus Enoploplastron 16% and Eodinium 11% of the population.     

Feeding response of a benthic copepod to ciliate prey type,prey concentration and habitat complexity

1. Protozoans are important consumers within microbial food webs and, in turn, they represent potential prey for small metazoans. However, feeding interactions within these food webs are rarely characterised and this is especially true for freshwater sediments. 2. We aimed to quantify the feeding links between a freshwater meiofaunal copepod and ciliates in two laboratory experiments. The first experiment addressed the response of Eucyclops serrulatus towards ciliate density and type (two ciliate species of the same genus differing in terms of body size). A second experiment assessed the effect of habitat structure on feeding rates by introducing different structural complexity into the feeding arena. In contrast to the first experiment, which was run only for one time period, this experiment also tested three different total feeding times (4, 7 and 9 h). 3. Eucyclops serrulatus exhibited high ingestion rates, with 3–69 ciliates copepod^?1 h^?1 consumed depending on food concentration, food type and habitat complexity. Copepods exhibited a preference for the smaller ciliate when total ciliate concentration was low, but selected both ciliates equally when food concentrations were medium or high. However, at very high food concentration, Eucyclops preferred the larger ciliate (which was 1/3 of its own body size), suggesting that the longer handling times of the larger prey are rewarding when the large prey is present in high numbers. In terms of total numbers consumed, copepods fed on more small ciliates, but in terms of carbon units both ciliates were selected equally when total prey concentration was low or medium. However, copepods derived more carbon from the larger prey at high and very high prey concentrations (up to 0.7 μgC out of a maximum of 1.1 μgC copepod^?1 h^?1). Habitat complexity influenced the feeding of copepods when it was observed over time. 4. The copepod–ciliate link is well known from the pelagic zone of both marine and freshwater habitats. We have shown its potential importance within the benthos, where it can be influenced by food identity, food quantity and possibly by habitat complexity.     

Molecular phylogeny and taxonomic revision of the genus Wittrockiella (Pithophoraceae,Cladophorales), including the descriptions of W. australis sp. nov. and W. zosterae sp. nov.

Wittrockiella is a small genus of filamentous green algae that occurs in habitats with reduced or fluctuating salinities. Many aspects of the basic biology of these algae are still unknown and the phylogenetic relationships within the genus have not been fully explored. We provide a phylogeny based on three ribosomal markers (ITS, LSU, and SSU rDNA) of the genus, including broad intraspecific sampling for W. lyallii and W. salina, recommendations for the use of existing names are made, and highlight aspects of their physiology and life cycle. Molecular data indicate that there are five species of Wittrockiella. Two new species, W. australis and W. zosterae, are described, both are endophytes. Although W. lyallii and W. salina can be identified morphologically, there are no diagnostic morphological characters to distinguish between W. amphibia, W. australis, and W. zosterae. A range of low molecular weight carbohydrates were analyzed but proved to not be taxonomically informative. The distribution range of W. salina is extended to the Northern Hemisphere as this species has been found in brackish lakes in Japan. Furthermore, it is shown that there are no grounds to recognize W. salina var. kraftii, which was described as an endemic variety from a freshwater habitat on Lord Howe Island, Australia. Culture experiments indicate that W. australis has a preference for growth in lower salinities over full seawater. For W. amphibia and W. zosterae, sexual reproduction is documented, and the split of these species is possibly attributable to polyploidization.     

Identification of Intracellular Bacteria in the Ciliate Balantidium ctenopharyngodoni (Ciliophora,Litostomatea)

The ciliate Balantidium ctenopharyngodoni is the most prominent protist in the guts of grass carp, where it mainly inhabits the creamy luminal contents of the hindgut. Ciliates are generally colonized by microorganisms via phagotrophic feeding. In order to study the intracellular bacteria in this ciliate, we have successfully established it in in vitro culture. Herein, we investigated and compared the bacterial community structures of cultured and freshly collected B. ctenopharyngodoni. The results showed that these two groups exhibited different bacterial communities. The most abundant bacterial family in freshly collected samples was Enterobacteriaceae, while in cultured samples it was Fusobacteriaceae. In addition, a key intracellular bacterium, Cetobacterium somerae, was identified in the cytoplasm of cultured ciliates using fluorescence in situ hybridization (FISH). This study shows that ciliates can retain the intracellular bacteria acquired in the natural habitat for quite a long time, but the bacterial community structure of ciliates eventually changes after a long period of cultivation.     

Morphology and Morphogenesis of Fuscheria terricola n. sp. and Spathidium muscorum (Ciliophora: Kinetofragminophora)1

The morphology and morphogenesis of the kinetofragminophoran soil ciliates, Fuscheria terricola n. sp. and Spathidium muscorum Dragesco & Dragesco-Kerneis, 1979, are described. Stained specimens (protargol) are characterized biometrically. The new species differs from the other species of the genus in its body size, body shape, number of kineties, length of extrusomes, and habitat. Both species have telokinetal stomatogenesis, which commences with a proliferation of kinetosomes at those kineties which bear the brosse. Fuscheria terricola does not have a complex perioral ciliature; indeed, it might be that this species has only monokinetids. Thus only a proliferation of kinetosomes and the separation of the kineties takes place in the prospective division furrow. In contrast, S. muscorum differentiates short dikinetid kinetofragments in the region of the division furrow, which are arranged to form the perioral kinety of the opisthe in the intermediate and late stages of the stomatogenesis. The right part of the perioral kinety develops first. This and other studies show that telokinetal stomatogenesis proceeds very differently depending on the differentiation of the oral ciliature; however, detailed studies on the morphogenesis of kinetofragminophoran ciliates are still too few in number for subtypes to be defined.     

Species of Gonostomum and Paragonostomum (Ciliophora, Hypotrichida, Oxytrichidae) from the Valley of Flowers, India, with descriptions of Gonostomum singhii sp nov, Paragonostomum ghangriai sp nov and Paragonostomum minuta sp nov

Soil samples taken from the Valley of Flowers, a component of the Nanda Devi Biosphere Reserve in the Himalayan regions of India showed the presence of twenty two free living species of ciliates. There is a preponderance of species which exhibit oral ciliature and ontogenesis in the Gonostomum pattern. Of the four species of the genus Gonostomum, three viz., G affine, G gonostomoida and G kuehnelti are similar to described species; Gonostomum singhii is new. The two species of genus Paragonostomum viz., P minuta and P ghangriai are new. The three new species are described in the present paper. All these species show prominent hypertrophied ciliary structures. Their paroral membranes reveal characteristic differences with respect to their position, number of constituent cilia and the distance between adjacent cilia. It is proposed that such species specific features of the paroral membrane have a bearing in exercising different food organism preferences as they co-exist at many sites. This single factor has possibly played an important role in species diversification of this group of hypotrichs in this isolated habitat.     

Infection of algae-free Climacostomum virens with symbiotic Chlorella sp. isolated from algae-containing C. virens

The ciliate Climacostomum virens normally contains algae as symbionts in its cytoplasm and retains them over many generations. An aposymbiotic strain of C. virens which cannot re-establish a new symbiotic association by ingestion of algae derived from green Climacostomum was recently isolated in our laboratory. Results of infection experiments showed that all newly ingested, potentially symbiotic algae were digested in food vacuoles. To clarify whether these ciliates have completely lost their ability to sustain symbiosis with algae or whether this ability can eventually be re-established, infection experiments were performed using a microinjection technique. We have achieved successful infection of algae-free Climacostomum using this method. The endosymbiont population was established in ciliates from as few as 3-5 injected algae, which have retained an intact perialgal vacuole membrane around them. To our knowledge, this is the first evidence of successful infection of aposymbiotic ciliates with algae by microinjection.     

Protozoan epibionts on Mysis relicta Loven, 1862 (Crustacea, Mysidacea) from Lake Lüšiai (Lithuania)

Protozoan epibionts were found on specimens of the mysid Mysis relicta collected in Lake Lü?iai (Lithuania). These protozoans belong to three genera of ciliates: Vorticella, Dendrosoma and Tokophrya, and were located on the antennae, carapace, appendages and abdomen of the basibiont. The ciliates of the genus Vorticella had a bell‐shaped body, with concave‐type striations and a very long contractile stalk. The stalk joined the body via a truncated and overlapping zone. The macronucleus was J‐shaped, and the contractile vacuole was anterolateral. Only one ciliate species of Vorticella has been found on mysids, although on species other than M. relicta, and this is therefore the first record of Vorticella on M. relicta, and the second on the order Mysidacea. The ciliates of the genus Dendrosoma had an irregular body directly attached to the substrate. Finger‐like actinophores arose from the body, with a bundle of tentacles at their ends. The macronucleus ramified into branches that followed the actinophores. These ciliates were included in the species D. astaci. The presence of this ciliate represents the first record of this genus on Mysis relicta. The ciliates of the genus Tokophrya had a pyramidal body, the apical surface of which showed two conspicuous actinophores, with a short, longitudinally striated stalk joining to the body via a narrow unstriated band. The macronucleus was spherical or ovoid, centrally located, and there was an apical contractile vacuole. Only one previous study has referred to the presence of this genus on Mysis relicta, and the data contained in it were compared with those of the present study. The biometrical data and taxonomical position of the ciliate epibionts was considered.     

Cochliopodium arabianum n. sp. (Amorphea,Amoebozoa)

A new species of Cochliopodium isolated from freshwater at Arabia Lake in Lithonia, GA, USA is described based on light microscopic morphology, fine structure, and molecular genetic evidence. Cochliopodium arabianum n. sp., previously labeled as “isolate Con1” in prior publications, has been shown to group within the genus Cochliopodium in our molecular phylogenetic analysis. Light microscopy and fine structure evidence indicates the new isolate not only shares characters of the genus but also unique distinctive features. Cochliopodium arabianum n. sp. is typically round when stationary; or oval to sometimes broadly flabellate or triangular in shape during locomotion, with average length of 35 μm and breadth of 51 μm. Fine structure evidence indicates C. arabianum n. sp. has tower‐like scales, lacking a terminal spine, sharing high similarity with its closest relative C. actinophorum. However, the scales of C. arabianum n. sp. are unique in height and the breadth of the base plate. Both morphological and molecular data, including SSU‐rDNA and COI, indicate that this new species falls in a clade sufficiently different from other species to suggest that it is a valid new species.     

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.     

Pelagic Ciliates in a Large Mesotrophic Lake: Seasonal Succession and Taxon‐Specific Bacterivory in Lake Constance

The taxonomic composition of the ciliate assemblage and their taxon‐specific bacterial grazing rates in Lake Constance were investigated over the course of one year. Bacterial grazing rates were measured using natural fluorescently labelled bacteria (FLB) and compared to bacterial production. Small species such as Balanion planctonicum/Urotricha furcata and Rimostrombidium spp./Halteria sp. were the most numerous ciliates on the annual average. Larger ciliates such as Rimostrombidium lacustris and Limnostrombidium spp. contributed significantly to total ciliate biomass, but were relatively unimportant as bacterial grazers. Per capita ingestion rates ranged from 0–194 bacteria ciliate⁻¹ h⁻¹ and changed seasonally up to a hundredfold within a given taxon. Approximately 1% of the bacterial production were removed by the ciliate community on the annual average. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

A mechanism of transmission and factors affecting coral susceptibility to <Emphasis Type="Italic">Halofolliculina</Emphasis> sp. infection

Anecdotal evidence collected since 2004 suggests that infections caused by ciliates in the genus Halofolliculina may be related to coral mortality in more than 25 scleractinian species in the Caribbean. However, the relationship between the presence of ciliates and coral mortality has not yet been firmly established. Field and laboratory manipulations were used to test if ciliate infections harm corals, if ciliates are able to infect healthy colonies, and if coral susceptibility to ciliate infection depends on temperature, depth, distance to an infected colony, and the presence of injuries. Ciliate infections were always characterized by a visually detectable front of ciliates located on recently exposed coral skeletons. These infections altered the normal structure of the colony by causing tissue mortality (0.8 ± 0.95 cm month⁻¹, mean ± SD) and by delaying or preventing recovery from injuries. Under laboratory conditions, ciliates transmitted directly and horizontally from infected to healthy hosts, and coral susceptibility to ciliate infections increased with the presence of injuries. After invasion, the ciliate population grew, rapidly and after 8 d, produced tissue mortality on 32% of newly infected hosts. Thus, our results support the existence of a new Caribbean coral syndrome that is associated with tissue mortality, is infectious, and transmits directly and horizontally. Even though the role of ciliates in the development of lesions on coral tissues remains unclear, their presence is by far the most conspicuous sign of this syndrome; thus, we propose to name this condition Caribbean ciliate infection (CCI). Communicated by Biology Editor Dr Michael Lesser     

Ultrastructure and Molecular Phylogeny of Mesodinium coatsi sp. nov., a Benthic Marine Ciliate

Mesodinium is a globally distributed ciliate genus forming frequent and recurrent blooms in diverse marine habitats. Here, we describe a new marine species, Mesodinium coatsi n. sp., originally isolated from interstitial water of surface sand samples collected at Mohang Beach, Korea. The species was maintained under a mixotrophic growth condition for longer than 1 yr by providing a cryptomonad, Chroomonas sp., as the sole prey. Cell morphology and subcellular structure were examined by light microscopy, scanning, and transmission electron microscopy, and molecular phylogeny was inferred from nuclear‐encoded 18S rDNA sequence data. Like other Mesodinium species, M. coatsi consisted of two hemispheres separated by two types of kinetids, and had tentacles located at the oral end of the cell. Several food vacuoles were observed in the cytoplasm, and partially digested prey cells sometimes existed in food vacuoles. Kinetids and the associated accessory structures were quite similar to those previously reported, but M. coatsi was differentiated from other marine Mesodinium species by ultrastructural characters of the dikinetids, polykinetids, and tentacles. We also provided a detailed illustration of infraciliature. Molecular phylogeny revealed that M. coatsi and Mesodinium chamaeleon were closely related to each other.