Thiago da Silva Paiva

An Antarctic hypotrichous ciliate, Parasterkiella thompsoni (Foissner) nov. gen., nov. comb., recorded in Argentinean peat-bogs: morphology, morphogenesis, and molecular phylogeny.

The ciliate Parasterkiella thompsoni (Foissner, 1996) nov. gen., nov. comb. was originally described from Antarctica. In the present study, we report the morphology, morphogenesis during cell division, and molecular phylogeny inferred from the 18S-rDNA sequence of a population isolated from the Rancho Hambre peat bog, Tierra del Fuego Province (Argentina). The study is based on live and protargol-impregnated specimens. Molecular phylogeny was inferred from trees constructed by means of the maximum parsimony, neighbor joining, and Bayesian analyses. The interphase morphology matches the original description of the species. During the cell division, stomatogenesis begins with the de novo proliferation of two fields of basal bodies, each one left of the postoral ventral cirri and of transverse cirri, which later unify. Primordia IV-VI of the proter develop from disaggregation of cirrus IV/3, while primordium IV of the opisthe develops from cirrus IV/2 and primordia V and VI from cirrus V/4. Dorsal morphogenesis occurs in the Urosomoida pattern-that is, the fragmentation of kinety 3 is lacking. Three macronuclear nodules are generated before cytokinesis. Phylogenetic analyses consistently placed P. thompsoni within the stylonychines. New data on the morphogenesis of the dorsal ciliature justifies the transference of Sterkiella thompsoni to a new genus Parasterkiella.

Morphology and 18S rDNA phylogeny of Hemicycliostyla sphagni (Ciliophora, Hypotricha) from Brazil with redefinition of the genus Hemicycliostyla

Morphology of the urostylid ciliate Hemicycliostyla sphagni Stokes, 1886, the type of Hemicycliostyla Stokes, 1886, is investigated based on live and protargol-impregnated specimens from a Brazilian population. The absence of transverse cirri, which has been considered the main diagnostic feature of Hemicycliostyla, separating it from Pseudourostyla Borror, 1972, was found to vary within the studied population, with 50% of the specimens exhibiting inconspicuous and/or rudimentary transverse cirri. A redefinition of Hemicycliostyla was possible based on combined features of interphase and divisional morphogenesis: Retroextendia Berger, 2006, with bi- or multicoronal frontal cirral pattern; fronto-terminal cirri present; multiple left and right marginal cirral rows that replicate independently via within-row development, each parental row producing one primordium per divider; caudal cirri lacking; and presence/absence of transverse cirri may be intrapopulationally variable. Phylogenetic analyses of the 18S rDNA marker unambiguously placed H. sphagni as sister group of Pseudourostyla franzi Foissner, 1987, which is herein transferred to Hemicycliostyla as Hemicycliostyla franzi comb. nov.

Phylogenetic study of Class Armophorea (Alveolata, Ciliophora) based on 18S-rDNA data

The 18S rDNA phylogeny of Class Armophorea, a group of anaerobic ciliates, is proposed based on an analysis of 44 sequences (out of 195) retrieved from the NCBI/GenBank database. Emphasis was placed on the use of two nucleotide alignment criteria that involved variation in the gap-opening and gap-extension parameters and the use of rRNA secondary structure to orientate multiple-alignment. A sensitivity analysis of 76 data sets was run to assess the effect of variations in indel parameters on tree topologies. Bayesian inference, maximum likelihood and maximum parsimony phylogenetic analyses were used to explore how different analytic frameworks influenced the resulting hypotheses. A sensitivity analysis revealed that the relationships among higher taxa of the Intramacronucleata were dependent upon how indels were determined during multiple-alignment of nucleotides. The phylogenetic analyses rejected the monophyly of the Armophorea most of the time and consistently indicated that the Metopidae and Nyctotheridae were related to the Litostomatea. There was no consensus on the placement of the Caenomorphidae, which could be a sister group of the Metopidae + Nyctorheridae, or could have diverged at the base of the Spirotrichea branch or the Intramacronucleata tree.The 18S rDNA phylogeny of Class Armophorea, a group of anaerobic ciliates, is proposed based on an analysis of 44 sequences (out of 195) retrieved from the NCBI/GenBank database. Emphasis was placed on the use of two nucleotide alignment criteria that involved variation in the gap-opening and gap-extension parameters and the use of rRNA secondary structure to orientate multiple-alignment. A sensitivity analysis of 76 data sets was run to assess the effect of variations in indel parameters on tree topologies. Bayesian inference, maximum likelihood and maximum parsimony phylogenetic analyses were used to explore how different analytic frameworks influenced the resulting hypotheses. A sensitivity analysis revealed that the relationships among higher taxa of the Intramacronucleata were dependent upon how indels were determined during multiple-alignment of nucleotides. The phylogenetic analyses rejected the monophyly of the Armophorea most of the time and consistently indicated that the Metopidae and Nyctotheridae were related to the Litostomatea. There was no consensus on the placement of the Caenomorphidae, which could be a sister group of the Metopidae + Nyctorheridae, or could have diverged at the base of the Spirotrichea branch or the Intramacronucleata tree.

Redescription of Licnophora chattoni Villeneuve-Brachon, 1939 (Ciliophora, Spirotrichea), associated with Zyzzyzus warreni Calder, 1988 (Cnidaria, Hydrozoa)

Licnophora chattoni, found in association with Zyzzyzus warreni, a tubulariid hydroid epizoic in sponges from São Sebastião (SP, Brazil), is redescribed and illustrated using light and electron microscopy. The ciliate has a flexible, transparent body formed by an oval anterior region linked to the posterior basal disc via a flexible neck region. Numerous cortical granules are observed scattered throughout the body and densely packed along the neck. The adoral zone is formed by about 81 external and 24 infundibular paramembranelles. The paroral membrane, formed by a row of long cilia arranged in monokinetids, extends through a groove in the body to the adhesive disc. Two dorsal kinetids are present along the right body margin and around the neck. The adhesive disc (18μm in diameter) lacks cilia in the area above the velum. The velum covers a row of dikinetids bearing long cilia and four dikineties, two or three of which are interrupted on the ventral surface. Nine to twelve macronuclear nodules connected by isthmuses are distributed in the cytoplasm, plus two nodules located in the adhesive disc and between those there is an ovate micronucleus.

Expanded phylogenetic analyses of the class Heterotrichea (Ciliophora, Postciliodesmatophora) using five molecular markers and morphological data.

Most studies of the molecular evolution of Heterotrichea have been based solely on the 18S-rDNA gene, which were inconsistent with morphological classification. Because of the limitations of single locus phylogenies and the recurring problem of lack of resolution of deeper nodes found in previous studies, we present hypotheses of the evolution of internal groups of the class Heterotrichea based on multi-loci analyses (18S-rDNA, 28S-rDNA, ITS1-5.8S-ITS2 region, COI and alpha-tubulin) and morphological data. Phylogenetic trees from protein coding gene data are presented for Heterotrichea for the first time. Phylogenetic analyses included Bayesian inference, maximum likelihood, maximum parsimony methods, and optimal trees were statistically compared to alternative topologies from the literature. Additionally, the Bayesian concordance approach (BCA algorithm) was used to assess the concordance factor between topologies obtained from isolated analyses. Because different loci may evolve at different rates, resulting in different gene topologies, we also estimated a species tree for Heterotrichea using the STAR coalescence-based method. The results show that: (1) single gene trees are inconsistent regarding the position of some heterotrichean families; (2) the concatenation of all data in a total-evidence tree improved the resolution of deep nodes among the heterotrichean families and genera; (3) the coalescent-based species tree is consistent with phylogenies based on the 18S-rDNA gene and shows Spirostomidae as the stem group of Heterotrichea; (4) however, the total-evidence tree suggests that the large Heterotrichea cluster is divided into nine lineages in which Peritromidae diverges at the base of the Heterotrichea tree.

Fine Structure and Molecular Phylogeny of Parametopidium circumlabens (Ciliophora: Armophorea), Endocommensal of Sea Urchins

Metopid armophoreans are ciliates commonly found in anaerobic environments worldwide; however, very little is known of their fine structure. In this study, the metopid Parametopidium circumlabens (Biggar and Wenrich 1932) Aescht, 1980, a common endocommensal of sea urchins, is investigated for the first time with emphasis on transmission electron microscopy, revealing several previously unknown elements of its morphology. Somatic dikinetids of P. circumlabens have a typical ribbon of transverse microtubules, an isolated microtubule near triplets 4 and 5 of the anterior kinetosome, plus two other microtubules between anterior and posterior kinetosomes, a short kinetodesmal striated fiber and long postciliary microtubules. In the dikinetids of the perizonal stripe, the kinetodesmal fiber is very pronounced, and there is a conspicuous microfibrillar network system associated with the kinetosomes. A new structure, shaped as a dense, roughly cylindrical mass surrounded by microtubules, is found associated with the posterior kinetosome of perizonal dikinetids. The paroral membrane is diplostichomonad and the adoral membranelles are of the “paramembranelle” type. Bayesian inference and maximum‐likelihood analysis of the 18S‐rDNA gene unambiguously placed P. circumlabens as sister group of the cluster formed by ((Atopospira galeata, Atopospira violacea) Metopus laminarius) + Clevelandellida, corroborating its classification within the Metopida.

Description and Phylogeny of Urostyla grandis wiackowskii subsp. nov. (Ciliophora, Hypotricha) from an Estuarine Mangrove in Brazil

Interphase specimens, aspects of physiological reorganization and divisional morphogenesis were investigated in a strain of a hypotrichous ciliate highly similar to Urostyla grandis Ehrenberg, (type species of Urostyla), collected from a mangrove area in the estuary of the Paraíba do Sul river (Rio de Janeiro, Brazil). The results revealed that albeit interphase specimens match with the known morphologic variability in U. grandis, morphogenetic processes have conspicuous differences. Parental adoral zone is entirely renewed during morphogenesis, and marginal cirri exhibit a unique combination of developmental modes, in which left marginal rows originate from multiple anlagen arising from innermost left marginal cirral row, whereas right marginal ciliature originates from individual within‐row anlagen. Based on such characteristics, a new subspecies, namely U. grandis wiackowskii subsp. nov. is proposed, and consequently, U. grandis grandis Ehrenberg, stat. nov. is established. Bayesian and maximum‐likelihood analyses of the 18S rDNA unambiguously placed U. grandis wiackowskii as adelphotaxon of a cluster formed by other U. grandis sequences. The implications of such findings to the systematics of Urostyla are discussed.

Morphology of Parastrongylidium estevesi comb. nov. and Deviata brasiliensis sp. nov. (Ciliophora: Stichotrichia) from a sewage treatment plant in Rio de Janeiro, Brazil

In samples of raw sewage collected from a sewage treatment plant in Rio de Janeiro (ETE-Penha), we found populations of two species, Deviata estevesi Paiva & Silva-Neto, 2005 and Deviata brasiliensis sp. nov. The organisms were studied in vivo under phase contrast microscopy, differential interference contrast (DIC), and after protargol-impregnation. The population of D. estevesi exhibited more extensive variation in cirral pattern than previously described. The interphasic organisms of new species D. brasiliensis sp. nov. are distinguishable from their congeners based on a series of morphometric features: cirral row R3 usually presents 1-3 cirri behind the right frontal cirrus, on average there are four macronuclear nodules, and, during morphogenesis of cell division, primordium V of the proter originates from the anterior region of cirral row R5 instead of row R6, as in the type species D. abbrevescens Eigner, 1995. In D. estevesi, the ventral cirral rows replicate by within-row primordia, which develop independently for the proter and for the opisthe, suggesting that it belongs to or is closely related to Parastrongylidium, hence the combination P. estevesi comb. nov. is established.

Description and Phylogeny of Tetrakeronopsis silvanetoi gen. nov., sp. nov. (Hypotricha, Pseudokeronopsidae), a New Benthic Marine Ciliate from Brazil

Pseudokeronopsidae Borror & Wicklow, 1983 are biotechnologically important ciliate protists which produce toxic defense substances; however, their diversity is still little known in Brazil. In the present study, Tetrakeronopsis silvanetoi, a new genus and species of marine pseudokeronopsid hypotrichs is described from samples of water with bottom sediment collected from the coast of São Paulo state. Its phylogenetic affinities to the “core urostyloids” are hypothesized based on analyses of the 18S-rDNA marker, and a new subfamily, the Nothoholostichinae subfam. nov., is erected to name the monophylum composed of pseudokeronopsids in which the anterior corona is usually formed by four frontal cirri. In addition, the new combination Monocoronella longissima comb. nov. is proposed for Nothoholosticha longissima (Dragesco & Dragesco-Kernéis, 1986) Li et al., 2009.

Morphology and divisional morphogenesis of Nudiamphisiella interrupta Foissner, Agatha & Berger, 2002 (Ciliophora: Stichotrichia) based on a Brazilian strain.

The morphology and divisional morphogenesis of a Brazilian strain of Nudiamphisiella interrupta were characterized. The organisms were investigated after protargol impregnation. Interphase cells of N. interrupta fitted the original description. Stomatogenesis in dividing cells occurred apokinetally, left of the amphisiellid cirral row, and the ventral ciliature was derived from five ventral primordia; primordium IV was absent. The dorsal kineties replicated intrakinetally, with the exception of kinety 4, which is interpreted as a dorso-marginal row, since it originates in association with the right marginal row. The nuclear apparatus divided as usual in non-urostylid stichotrichs, exhibiting complete fusion of the macronuclear nodules in middle dividers.