Seasonal succession of ciliate Mesodinium spp. with red, green, or mixed plastids and their association with cryptophyte prey.

Mesodinium spp. are commonly found in marine and brackish waters, and several species are known to contain red, green, or both plastids that originate from cryptophyte prey. We observed the seasonal succession of Mesodinium spp. in a Japanese brackish lake, and we analysed the origin and diversity of the various coloured plastids within the cells of Mesodinium spp. using a newly developed primer set that specifically targets the cryptophyte nuclear 18S rRNA gene. Mesodinium rubrum isolated from the lake contained only red plastids originating from cryptophyte Teleaulax amphioxeia. We identified novel Mesodinium sp. that contained only green plastids or both red and green plastids originating from cryptophytes Hemiselmis sp. and Teleaulax acuta. Although the morphology of the newly identified Mesodinium sp. was indistinguishable from that of M. rubrum under normal light microscopy, phylogenetic analysis placed this species between the M. rubrum/major species complex and a well-supported lineage of M. chamaeleon and M. coatsi. Close associations were observed in cryptophyte species composition within cells of Mesodinium spp. and in ambient water samples. The appearance of suitable cryptophyte prey is probably a trigger for succession of Mesodinium spp., and the subsequent abundance of Mesodinium spp. appears to be influenced by water temperature and dissolved inorganic nutrients.

Euduboscquella costata n. sp. (Dinoflagellata, Syndinea), an Intracellular Parasite of the Ciliate Schmidingerella arcuata: Morphology, Molecular Phylogeny, Life Cycle, Prevalence, and Infection Intensity.

The syndinean dinoflagellate Euduboscquella costata n. sp., an intracellular parasite of the tintinnid ciliate Schmidingerella arcuata, was discovered from Korean coastal water in November of 2013. Euduboscquella costata parasitized in about 62% of the host population, with infection intensity (= number of trophonts in a single host cell) ranging from 1 to 8. Based on morphology and nuclear 18S ribosomal RNA gene sequences, the parasite is new to science. Euduboscquella costata n. sp. had an infection cycle typical of the genus, but had morphological and developmental features that distinguished it from congeneric species. These features include: (1) episome of the trophont with 25-40 grooves converging toward the center of the shield; (2) a narrow, funnel-shaped lamina pharyngea extending from the margin of the episomal shield to the nucleus; (3) persistence of grooves during extracellular development (sporogenesis); (4) a single food vacuole during sporogenesis; (5) separation of sporocytes early in sporogenesis, regardless of type of spore formed; and (6) dinospore size (ca. 14 µm in length) and shape (bulbous episome with narrower, tapering hyposome). After sporogenesis, E. costata produced four different types of spore that showed completely identical 18S rRNA gene sequences. The gene sequence was completely identical with a previously reported population, Euduboscquella sp. ex S. arcuata, from Assawoman Bay, USA, indicating that the two populations are likely conspecific. Favella ehrenbergii, a widely recorded tintinnid known to host Euduboscquella spp., co-occurred with S. arcuata, but was not infected by E. costata in field samples or during short-term, cross-infection experiments.

Effects of a ciliate protozoa predator on microbial communities in pitcher plant (Sarracenia purpurea) leaves.

The aquatic communities found within the water filled leaves of the pitcher plant, Sarracenia purpurea, have a simple trophic structure providing an ideal system to study microscale interactions between protozoan predators and their bacterial prey. In this study, replicate communities were maintained with and without the presence of the bactivorous protozoan, Colpoda steinii, to determine the effects of grazing on microbial communities. Changes in microbial (Archaea and Bacteria) community structure were assessed using iTag sequencing of 16S rRNA genes. The microbial communities were similar with and without the protozoan predator, with>1000 species. Of these species, Archaea were negligible, with Bacteria comprising 99.99% of the microbial community. The Proteobacteria and Bacteroidetes were the most dominant phyla. The addition of a protozoan predator did not have a significant effect on microbial evenness nor richness. However, the presence of the protozoan did cause a significant shift in the relative abundances of a number of bacterial species. This suggested that bactivorous protozoan may target specific bacterial species and/or that certain bacterial species have innate mechanisms by which they evade predators. These findings help to elucidate the effect that trophic structure perturbations have on predator prey interactions in microbial systems.

Frequency and biodiversity of symbionts in representatives of the main classes of Ciliophora.

Representatives of all classes of Ciliophora have been studied for the detection and investigation of both prokaryotic and eukaryotic (not algal) endo- (EnS) and ectosymbionts (EcS). Different methods including transmission electron microscopy (TEM) and fluorescence in situ hybridisation (FISH) have been used. Apparently, the capability of keeping symbionts varies among the different ciliate groups as it generally is the case in different protist taxa. Most of the prokaryotic EnSs detected belong to Alphaproteobacteria. Holospora or Holospora-like infectious bacteria of this group were found in representatives of Heterotrichea, Armophorea, Phyllopharyngea, Prostomatea and mainly of Oligohymenophorea. Bacteria associated with bacteriophages were found in species of Heterotrichea and Oligohymenophorea. This holds true also for bacteria with R-bodies. A quite rare type of EnS - motile bacteria - was found in ciliates of the same two classes as well, either in the cytoplasm (Heterotrichea) or in the macronucleus and its perinuclear space (Oligohymenophorea). EcSs are more common in Heterotrichea, Armophorea and Plagiopylea, but were never found in other groups. Among the eukaryotic EnSs of ciliates, very few representatives of Microsporidia and Trypanosomatidae were recorded. In conclusion, heterotrichs and oligohymenophoreans are the most promising groups of Ciliophora for the investigation of symbiosis.

Revision of the family Duboscquellidae with description of Euduboscquella crenulata n. gen., n. sp. (Dinoflagellata, Syndinea), an intracellular parasite of the ciliate Favella panamensis Kofoid & Campbell.

Recent recognition that tintinnids are infected by dinophycean as well as syndinean parasites prompts taxonomic revision of dinoflagellate species that parasitize these ciliates. Long overlooked features of the type species Duboscquella tintinnicola are used to emend the genus and family Duboscquellidae, resulting in both taxa being moved from the Syndinea to the Dinophyceae. Syndinean species previously classified as Duboscquella are relocated to Euduboscquella n. gen., with Euduboscquella crenulata n. sp. as the type. As an endoparasitic species, E. crenulata shares with its congeners processes associated with intracellular development and sporogenesis, but differs from closely related species in nuclear and cortical morphology of the trophont, including a distinctively grooved shield (= episome) that imparts a crenulated appearance in optical section. In addition, E. crenulata produces three morphologically distinct spore types, two of which undergo syngamy to form a uninucleate zygote. The zygote undergoes successive division to produce four daughter cells of unequal size, but that resemble the nonmating spore type.

New records of the ectoparasitic flagellate Colpodella gonderi on non-Colpoda ciliates.

Colpodella gonderi is the only ectoparasitic flagellate of ciliated protozoa described thus far. This investigation reveals new records of C. gonderi retrieved from soil samples in southern Scotland, UK. Of fourteen ciliates species identified in one single occasion, three of them, Colpoda steinii, Pseudoplatyophrya nana and Grossglockneria acuta, were infested with the parasite. These results provide further evidence that C. gonderi is not host-specific of the ciliate genus Colpoda.

Tintinnophagus acutus n. g., n. sp. (Phylum Dinoflagellata), an ectoparasite of the ciliate Tintinnopsis cylindrica Daday 1887, and its relationship to Duboscquodinium collini Grassé 1952.

The dinoflagellate Tintinnophagus acutus n. g., n. sp., an ectoparasite of the ciliate Tintinnopsis cylindrica Daday, superficially resembles Duboscquodinium collini Grassé, a parasite of Eutintinnus fraknoii Daday. Dinospores of T. acutus are small transparent cells having a sharply pointed episome, conspicuous eyespot, posteriorly positioned nucleus with condensed chromosomes, and rigid form that may be supported by delicate thecal plates. Dinospores attach to the host via a feeding tube, losing their flagella, sulcus, and girdle to become spherical or ovoid cells. The trophont of T. acutus feeds on the host for several days, increasing dramatically in size before undergoing sporogenesis. Successive generations of daughter sporocytes are encompassed in an outer membrane or cyst wall, a feature not evident in trophonts. Tintinnophagus acutus differs from D. collini in host species, absence of a second membrane surrounding pre-sporogenic stages, and failure to differentiate into a gonocyte and a trophocyte at the first sporogenic division. Phylogenetic analyses based on small subunit (SSU) ribosomal DNA (rDNA) sequences placed T. acutus and D. collini in the class Dinophyceae, with T. acutus aligned loosely with Pfiesteria piscicida and related species, including Amyloodinium ocellatum, a parasite of fish, and Paulsenella vonstoschii, a parasite of diatoms. Dubosquodinium collini nested in a clade composed of several Scrippsiella species and Peridinium polonicum. Tree construction using longer rDNA sequences (i.e. SSU through partial large subunit) strengthened the placement of T. acutus and D. collini within the Dinophyceae.

Species of the parasitic genus Duboscquella are members of the enigmatic Marine Alveolate Group I.

Small subunit ribosomal RNA gene sequences of Duboscquella spp. infecting the tintinnid ciliate, Favella ehrenbergii, were determined. Two parasites were sampled from different localities. They are morphologically similar to each other and both resemble D. aspida. Nevertheless, two distinct sequences (7.6% divergence) were obtained from them. Phylogenetic trees inferred from maximum likelihood and maximum parsimony revealed that these two Duboscquella spp. sequences are enclosed in an environmental clade named Marine Alveolate Group I. This clade consists of a large number of picoplanktonic organisms known only from environmental samples from various parts of the ocean worldwide, and which therefore lack clear characterization and identification. Here, we provide morphological and genetic characterization of these two Duboscquella genotypes included in this enigmatic clade. Duboscquella spp. produce a large number of small flagellated spores as dispersal agents and the presence of such small cells partially explains why the organisms related to these parasites have been detected within environmental genetic libraries, built from picoplanktonic size fractions of environmental samples. The huge diversity of the Marine Alveolate Group I and the finding that parasites from different marine protists belong to this lineage suggest that parasitism is a widespread and ecologically relevant phenomenon in the marine environment.

Zooxanthellae harvested by ciliates associated with brown band syndrome of corals remain photosynthetically competent.

Brown band syndrome is a new coral affliction characterized by a local accumulation of yet-unidentified ciliates migrating as a band along the branches of coral colonies. In the current study, morphologically intact zooxanthellae (= Symbiodinium) were observed in great numbers inside the ciliates (>50 dinoflagellates per ciliate). Microscale oxygen measurements and variable chlorophyll a fluorescence analysis along with microscopic observations demonstrated that zooxanthellae within the ciliates are photosynthetically competent and do not become compromised during the progression of the brown band zone. Zooxanthellae showed similar trends in light acclimation in a comparison of rapid light curve and steady-state light curve measures of variable chlorophyll a fluorescence. Extended light exposure of steady-state light curves resulted in higher quantum yields of photosystem II. The brown band tissue exhibited higher photosynthetically active radiation absorptivity, indicating more efficient light absorption due to a higher density of zooxanthellae in the ciliate-dominated zone. This caused relatively higher gross photosynthesis rates in the zone with zooxanthella-containing ciliates compared to healthy coral tissue. The observation of photosynthetically active intracellular zooxanthellae in the ciliates suggests that the latter can benefit from photosynthates produced by ingested zooxanthellae and from photosynthetic oxygen production that alleviates diffusion limitation of oxic respiration in the densely populated brown band tissue. It remains to be shown whether the zooxanthellae form a stable symbiotic association with the ciliate or are engulfed incidentally during grazing on coral tissue and then maintained as active inside the ciliate for a period before being digested and replaced by new zooxanthellae.

A re-appraisal of the diversity of the methanogens associated with the rumen ciliates.

The diversity of methanogenic archaea associated with different species of ciliated protozoa in the rumen was analysed. Partial fragments of archaeal SSU rRNA genes were amplified from DNA isolated from single cells from the rumen protozoal species Metadinium medium, Entodinium furca, Ophryoscolex caudatus and Diplodinium dentatum. Sequence analysis of these fragments indicated that although all of the new isolates clustered with sequences previously described for methanogens, there was a difference in the relative distribution of sequences detected here as compared to that of previous work. In addition, many of the novel sequences, although clearly of archaeal origin have relatively low identity to the sequences in database which are most closely related to them.

Opioid activity of beta-endorphin-like proteins from Tetrahymena.

Morphine and other opioids have been reported to modulate phagocytosis in the ciliate Tetrahymena. However, the endogenous signaling molecule responsible for these effects remains uncharacterized. In this work we present evidence for the presence of beta-endorphin-like protein(s) in Tetrahymena thermophila. Subcellular extracts and cell-free culture supernatants were fractionated by hydrophobic chromatography on Sep Pack C18 columns and by affinity chromatography on polyclonal anti-beta-endorphin columns. Both preparations exhibited opioid-like effects in two different systems: 1) they inhibited phagocytosis in murine peritoneal macrophages, and 2) they blocked the response to mechanical stimuli in the ciliate Stentor. Both of these effects were reversed by naloxone, consistent with an opioid receptor-mediated mechanism. Chromatographic (HPLC) fractionation of the subcellular extracts resolved a component with beta-endorphin-like immunoreactivity, whose retention time was similar to that of the human beta-endorphin standard. Fractions were also analyzed by immunoblots using a monoclonal antibody that recognizes the N-terminus of human beta-endorphin. This antibody detected two antigenic components (corresponding to Mr 9,000 and Mr 12,000 polypeptides) in subcellular extracts, but only a single antigen (corresponding to a Mr 7,000 polypeptide) in culture supernatants. These results indicate that Tetrahymena produces one or more proteins that share some properties with beta-endorphin and that these may form part of an opioid mechanism that originated early in evolution.

Comportamento e distribuiçäo de protozoários ciliados (Protista, Ciliophora) no rúmen e no retículo de bovinos submetidos ao jejum / Protozoa ciliate (Protista, Ciliophora) behaviour and distribution in the rumen and reticulum of fasting bovines

Com o objetivo de verificar a distribuiçäo dos ciliados no rúmen e no retículo de bovinos e analiasar o possível comportamento de escape de isotriquídeos ao retículo, foram examinadas 18 amostras de conteúdo desses compartimentos, sendo nove de cada. As amostras foram obtidas de bovinos em jejum por, no mínimo, 12 horas, recém-abatidos no Matadouro Municipal de Juiz de Fora, MG, fixadas em formalina a 18,5 por cento e a quantificaçäo dos ciliados feita com o emprego da câmara Sedgewick-Rafter. Os gêneros dos ciliados observados, sua quantificaçäo e percentuais no rúmen e no retículo, respectivamente, foram os seguintes: Entodinium (10,01x10(5), 55,6 por cento; 79,84x10(4), 44,4 por cento), Diplodinium (12,32x10(4), 54,2 por cento; 10,40x10(4),45,4 por cento), Isotricha (14,40x10(4), 32 por cento; 30,56x10(4), 68 por cento), Dasytricha (25,12x10(4), 36,8 por cento; 43,20x10(4), 63,2 por cento), Eremoplastron (21,28x10(4), 77,3 por cento; 6,24x10(4), 22,7 por cento), Eudiplodinium (6,4x10(4), 48,2 por cento; 6,88x10(4), 51,8 por cento), Ostracodinium (14,24x10(4), 56 por cento; 11,20x10(4), 44 por cento), Eodinium (44,80x10(4), 58,2 por cento; 32,16x10(4), 41,8 por cento), Polyplastron (0,0; 0,16x10(4), 100 por cento), Diploplastron (2,8x10(4), 72 por cento; 1,12x10(4) 28 por cento), Epidinium (10,72x10(4), 54 por cento; 9,12x10(4), 46 por cento), Metadinium (4x10(4) 53,2 por cento; 3,52x10(4), 46,8 por cento) e Elytroplastron (0,64x10(4), 44,4 por cento; 0,80x10(4), 55,6 por cento). Houve diferença significativa no número total de ciliados e entre os gêneros Entodinium, Isotricha, Dasytricha, Eremoplastron, Eodinium e Diploplastron ocorrentes no rúmen e no retículo de bovinos nas condiçöes estudadas, destacando-se o maior número total de ciliados no rúmen e o maior número de isotriquídeos no retículo. Os resultados confirmam o escape de isotriquídeos do rúmen ao retículo, verificada pela predominância de isotricha (68 por cento) e de Dasytricha (63,2 por cento) neste compartimento após período de jejum, quando foram obtidas as amostras.

Presence of two suctorian ciliate species of the genera Acineta and Pelagacineta, epibionts on marine copepod crustaceans.

Acineta arcachoniensis sp. nov., a new species of protist suctorian epibiont on the copepod Chiridius gracilis is described. The individuals show two well-developed actinophores, with two fascicles of capitate tentacles, an irregular nucleus, a cup-shaped lorica and a short stalk. Reproduction of A. arcachoniensis is by endogenous budding. The suctorians are located on the base of the antennae, the final abdominal segments or on the last pereiopod pair of the copepod. In addition, a new registration of the suctorian Pelagacineta euchaetae on the copepod Calanus helgolandicus is recorded.

Phylogenetic study of methanogens associated with rumen ciliates.

The phylogeny of methanogenic archaea associated with ciliate protozoa in a sheep rumen was investigated. Ruminal ciliate protozoa were exhaustively washed and mixtures of genomic DNA extracted. Archaea-specific nested PCR amplification was conducted with the ciliate genomic mixture. The resultant small subunit (16S) ribosomal RNA gene (ssu rDNA) was cloned into Escherichia coli JM 109. Many methanogens were still observed on and/or in ciliate cells by fluorescent microscopy even after exhaustive washing with buffer. Partial sequences of ssu rDNA close to Methanobrevibacter smithii were dominant in the retrieved sequences. RFLP analyses on the retrieved sequences revealed the absence of Methanobrevibacter ruminantium in the protozoal preparation. The association of Methanobrevibacter spp. with ruminal ciliate protozoa was demonstrated by the isolation of archaeal ssu rDNA phylogenetically close to that of M. smithii.

Infraciliature, morphogenesis and life cycle of Endosphaera terebrans (Suctoria, Tokophridae).

The morphology, infraciliature, and life cycle of Endosphaera terebrans, a suctorian endocommensal of peritrichs, have been studied with the aid of silver impregnation. The life cycle of Endosphaera terebrans begins with infection of the host cell by a small larva. The swarmer has a pointed needle-like cellular projection and two rings of cilia. The swarmer penetrates the the peritrich, loses the cilia, and then matures into an adult. The infraciliature of the adult form has four rows of barren kinetosomes that lack kinetodesmal fibers. By endogenous budding, a migratory larva is produced that leaves the host cell through the peristomial disc and that can infect other peritrichs.

[New species of parasitic infusoria of the family Urceolariidae]. / Novye vidy paraziticheskikh infuzorii semeistva Urceolariidae.

Three new species of Ciliata of the family Urceolariidae were found as follows: Paratrichodina voikarensis sp. n. on the gills of Coregonus nasus and C. peled; P. uralensis sp. n. on the gills of the sterlet and Trichodina percae sp. n. in the nasal cavities of the perch. Figures and descriptions of the new species are given.

[Morphobiological characteristics of infusoria of the genus Chilodonella (Ciliata: Chlamydodontidae)]. / Morfobiologicheskie osobennosti infuzorii roda Chilodonella (Ciliata: Chlamydodontidae).

8 species of fishes from fishfarms of central and north--western regions of the USSR were examined. Chilodonella cyprini Moroff, 1902 and Ch. hexastichus Kiernik, 1909 were found. The reproductive process of infusoria was investigated. The data on the susceptibility of hosts of different species and age are given. The variability of Chilodonella depending on localisation and water temperature is shown.