A multigene timescale and diversification dynamics of Ciliophora evolution.

Ciliophora is one of the most diverse lineages of unicellular eukaryotes. Nevertheless, a robust timescale including all main lineages and employing properly identified ciliate fossils as primary calibrations is lacking. Here, we inferred a time-calibrated multigene phylogeny of Ciliophora evolution, and we used this timetree to investigate the rates and patterns of lineage diversification through time. We implemented a two-step analytical approach that favored both gene and taxon sampling, reducing the uncertainty of time estimates and yielding narrower credibility intervals on the ribosomal-derived chronogram. We estimate the origin of Ciliophora at 1143 Ma, which is substantially younger than previously proposed ages, and the huge diversity explosion occurred during the Paleozoic. Among the current groups recognized as classes, Spirotrichea diverged earlier, its origin was dated at ca. 850 Ma, and Protocruziea was the younger class, with crown age estimated at 56 Ma. Macroevolutionary analysis detected a significant rate shift in diversification dynamics in the spirotrichean clade Hypotrichia + Oligotrichia + Choreotrichia, which had accelerated speciation rate ca. 570 Ma, during the Ediacaran-Cambrian transition. For all crown lineages investigated, speciation rates declined through time, whereas extinction rates remained low and relatively constant throughout the evolutionary history of ciliates.

Resting Cysts of the Pigmented Ciliate Blepharisma sinuosum Sawaya, 1940 (Ciliophora: Heterotrichea).

The morphology of Blepharisma sinuosum resting cysts and the dynamics of pigmentation at different stages of encystment are presented for the first time. Cyst morphometrics are similar to other Blepharisma species, with three-wall layers, bacteria surrounding the ectocyst, a conical plug, and wrinkly surface toward the plug in mature stages. The vegetative moniliform macronucleus changes to a horseshoe shape, and at early stages, the cystic cytoplasm is homogeneously pigmented, comprising a contractile vacuole; later, pigments polarize toward the plug, decorate the cortical layer, and become brownish. This work reinforces the potential role of pigment dynamics on cyst biology.

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.

Ontogeny and Molecular Phylogeny of Apoamphisiella vernalis Reveal Unclear Separation between Genera Apoamphisiella and Paraurostyla (Protozoa, Ciliophora, Hypotricha).

Morphology and divisional morphogenesis of the hypotrich ciliate Apoamphisiella vernalis are investigated based on two populations from Brazil. Typical specimens of A. vernalis replicates its ventral ciliature from six fronto-ventral-transverse (FVT) anlagen independently formed for proter and opisthe, plus one or more short anlagen located between IV and V, which form surplus transverse cirri. Dorsal morphogenesis occurs as in typical oxytrichid dorsomarginalians, viz., with formation of three anlagen and fragmentation of the rightmost one. Dorsomarginal kineties are formed near anterior end of right marginal cirral row anlagen. Various anomalous specimens exhibiting more than two long ventral rows were found, which are explained by increasing the number of FVT anlagen and/or the number of cirri produced by anlagen. Comparative ontogeny and phylogenetic analyses based on the 18S rDNA reveal that Apoamphisiella vernalis is closely affine to North American and European strains of the Paraurostyla weissei complex. Their reduced genetic distances and conspicuous morphological variability show that both genera can overlap, which implies the necessity of re-evaluating the contextual relevance of some morphological characters commonly used for genus-level separation within hypotrich taxa.

Morphology and 18S rDNA gene sequence of Blepharisma sinuosum Sawaya, 1940 (Ciliophora: Heterotrichea) from Brazil.

The morphology and morphometric data of seven populations of Blepharisma sinuosum from southeastern Brazil were investigated. The description is based on live observations, protargol impregnation, and scanning electron microscopy. Blepharisma sinuosum measures 75-255µm in length and 25-93µm in width and has a spindle-shaped body, pink color, a single contractile vacuole located at the posterior end, 50 adoral membranelles, a conspicuous paroral, 17-35 somatic kineties, a moniliform macronucleus with 2-7 connected nodules, and 3-20 micronuclei. Morphological comparisons with similar species were performed and suggest that B. americanum is the junior synonym of B. sinuosum. The 18S rDNA gene sequence of B. sinuosum was obtained and compared with that of other Blepharisma species. The length and GC content of the obtained sequence is 1652bp and 47.03%, respectively, and has a very high structural similarity (99.9%) with the B. undulans sequence. The validity of the classification of Blepharisma species in morphonuclear subgenera is also discussed.

Occurrence of Trichodina heterodentata (Ciliophora: Trichodinidae) infesting tadpoles of Rhinella pombali (Anura: Bufonidae) in the Neotropical area.

Data on trichodinid ciliophorans in Brazil are scarce and generally do not use the modern silver-impregnation technique, which is essential to the taxonomy of the group. The present study reports not only on the first occurrence of Trichodina heterodentata in Brazil but also on the first record of this trichodinid infesting tadpoles of the species Rhinella pombali. This study also constitutes a second report on T. heterodentata from tadpoles. In September 2007, tadpoles were collected from a stream in a small farm in the agricultural area of the city of Juiz de Fora, southeastern Brazil. The ciliophorans found on the tadpoles' bodies and tails were submitted to techniques such as silver impregnation and scanning electron microscopy. Our biometric data of T. heterodentata population infesting Rhinella pombali were compared to five other South-African populations of T. heterodentata infesting tadpoles of Xenopus laevis laevis. Results were discussed in terms of low host specificity as well as the wide geographical distribution of this trichodinid.