Multivariate morphometric analyses of the predatory ciliate genus Semispathidium (Ciliophora: Litostomatea), with description of S. longiarmatum nov. spec.

We studied morphometrical variation, species boundaries, and importance of morphometric features for a reliable separation of five African Semispathidium taxa. Altogether, 20 features traditionally used in alpha-taxonomy of the predatory genus Semispathidium were measured or scored on 85 protargol-impregnated interphase specimens, and were analyzed using hierarchical clustering as well as principal component and canonical discriminant analyses. This multivariate approach confirmed that a population found in Botswanan floodplain soil represents a distinct taxon. The new species is described here as S. longiarmatum, using live observation, protargol impregnation, and scanning electron microscopy. Semispathidium longiarmatum strongly resembles S. armatum and S. breviarmatum but it is clearly distinguished from these species by the extrusome pattern. The reliability of S. longiarmatum is also strengthened, according to the canonical discriminant analysis, by several quantitative features, viz., the number of ciliary rows, the length:width ratio of the macronucleus, and the number of dikinetids in brush row 1. Moreover, the present study documents the distinctness of all African Semispathidium species which can be separated by a combination of both qualitative and quantitative (morphometric) features. Consequently, Semispathidium species do not form a continuous complex but fairly discrete clusters in the phenotypic space.     

Two new marine pleurostomatid ciliates from China,Amphileptus gui nov. spec. and Amphileptus yuianus nov. spec. (Ciliophora,Pleurostomatida)

The morphology and infraciliature of two pleurostomatid ciliates, Amphileptus gui nov. spec. and A. yuianus nov. spec., collected from mariculture ponds near Qingdao (Tsingtao), China, were investigated using living observations and the protargol impregnation method. These new species are distinguished from their congeners by a combination of characters including their marine habitat and the presence of an apical group of densely packed extrusomes. Amphileptus gui nov. spec. differs from A. yuianus nov. spec. mainly in the number of contractile vacuoles (3–7 vs. 1), the shape of extrusomes (thin bar-like vs. clavate), and the number of left and right somatic kineties (7–11 and 37–50 vs. 4 and 18–22).     

Synergistic effects of combining morphological and molecular data in resolving the phylogenetic position of Semispathidium (Ciliophora,Haptoria) with description of Semispathidium breviarmatum sp. n. from tropical Africa

We describe a new species, Semispathidium breviarmatum sp. n., from tropical Africa and analyse its phylogenetic position within the subclass Haptoria, using live observation, various silver impregnation methods, SEM and the 18S rRNA gene. Semispathidium breviarmatum differs from its congeners by the much higher number of ciliary rows and by the shape and size of the extrusomes, that is, extrusive organelles that kill the prey. The phylogenetic position of Semispathidium is controversial due to its ‘hybrid’ morphology. Specifically, the cylindroidal body has a more or less discoidal oral bulge indicating an enchelyodonid origin, while the anteriorly curved somatic kineties suggest a spathidiid ancestor. In order to reconstruct the evolutionary history of Semispathidium and to unravel its affinity to other haptorians, we used synergistic effects of combining morphological and molecular data coming from 34 haptorian taxa. These analyses show that Semispathidium belongs to the order Spathidiida representing a basal lineage that is far from ordinary Spathidium species, but very likely related to Protospathidium and Enchelys. Any closer phylogenetic relationship between Semispathidium and Enchelyodon spp. is not recognized in morphological and molecular phylogenies and is consistently excluded by statistical tree topology tests.     

Morphology and molecular phylogeny of two new soil ciliates,Hemiurosomoida warreni nov. spec. and Hemiurosoma clampi nov. spec. (Ciliophora,Hypotrichia) from Tibet

Two novel hypotrichous ciliates, Hemiurosomoida warreni nov. spec. and Hemiurosoma clampi nov. spec., isolated from soil in the Lhalu Wetland and Motuo Virgin Forest in Tibet, respectively, were investigated using live observation and protargol staining. Hemiurosomoida warreni nov. spec. strongly resembles the type species H. longa but can be distinguished by its body size in vivo (110–145 × 30–40 μm vs. 50–100 × 18–40 μm), number of adoral membranelles (25–38 vs. 15–22), and numbers of right (29–39 vs. 14–23) and left (26–35 vs. 13–23) marginal cirri, transverse cirri (3 vs. 4 or 5) and macronuclear nodules (4–8 vs. 2). Hemiurosoma clampi nov. spec. is characterized by its vermiform body shape, colourless cortical granules distributed in irregular rows, two macronuclear nodules, three frontal cirri, one buccal cirrus, four frontoventral cirri ranged in a line, two transverse cirri, lacking postoral ventral and pretransverse ventral cirri, and marginal rows that are not posteriorly confluent. Phylogenetic analyses based on SSU rDNA gene sequences suggest that Hemiurosomoida is not monophyletic. A close relationship is revealed between Hemiurosomoida warreni nov. spec., Parakahilella macrostoma, Hemiurosoma clampi nov. spec., and the type species Hemiurosoma terricola. As expected, all these species are classified within the “non-oxytrichid Dorsomarginalia”.     

Taxonomic studies on three marine pleurostomatid ciliates, Litonotus bergeri nov. spec., L. blattereri nov. spec. and L. petzi nov. spec. (Ciliophora, Pleurostomatida) from North China Sea

The morphology and infraciliature of three pleurostomatid ciliates, Litonotus bergeri nov. spec., L. blattereri nov. spec. and L. petzi nov. spec., collected from mariculture ponds near Qingdao (Tsingtao), China, were investigated using live observations and the protargol impregnation method. These new species are distinguished from their congeners by a combination of characters including the typical distribution of extrusomes, i.e., along entire ventral margin, the number of macronuclear nodules, features and number of somatic kineties, living morphology, number and position of contractile vacuoles and their marine biotopes. Considering the distribution of extrusomes and general morphology, five new combinations are suggested, Litonotus vermiforme (Sauerbrey, 1928) nov. comb. [basionym: Loxophyllum vermiforme Sauerbrey, 1928], Litonotus levigatum (Sauerbrey, 1928) nov. comb. [basionym: Loxophyllum levigatum Sauerbrey, 1928], Litonotus undulatum (Sauerbrey, 1928) nov. comb. [basionym: Loxophyllum undulatum Sauerbrey, 1928], Loxophyllum pictus (Gruber, 1884) nov. comb. [basionym: Litonotus pictus Gruber, 1884] and Loxophyllum trichocystiferus (Foissner, 1984) nov. comb. [basionym: Litonotus trichocystiferus Foissner, 1984].     

Pleospora drummondii spec. nov. und ihr Anamorph Stemphylium drummondii spec. nov.

Pleospora drammondii spec. nov. and its anamorph Stemphylium drammondii spec. nov. The causal organism of a leaf spot disease on Phlox drummondii is described as Pleospora drummondii spec. nov. and its anamorph Stempbylium drummondii spec, nov. — in contrast to earlier reports from the USA, in which the fungus was identified as S. botryosum.     

Description of two new marine pleurostomatid ciliates, Loxophyllum choii nov. spec. and L. shini nov. spec. (Ciliophora, Pleurostomatida) from China

The morphology and infraciliature of two pleurostomatid ciliates, Loxophyllum choii nov. spec. and L. shini nov. spec., collected from inshore waters and a shrimp-culturing pond near Qingdao (Tsingtao), China, were investigated using live observations and the protargol-impregnation method. L. choii is distinguished from its congeners by the presence of conspicuous warts and prominent spines along the dorsal margin. L. shini is identified by a combination of characters including the small body size, the presence of two macronuclear nodules and the evenly distributed extrusomes whose tips protrude from the surface along both margins.     

Characterization of Euplotes lynni nov. spec., E. indica nov. spec. and description of E. aediculatus and E. woodruffi (Ciliophora,Euplotidae) using an integrative approach

Four species belonging to the genus Euplotes have been investigated, namely: E. lynni nov. spec., E. indica nov. spec., E. aediculatus, and E. woodruffi. All populations are from India and were investigated using morphological and molecular markers. The phylogenetic relationships were inferred from small subunit ribosomal rRNA gene (SSU rRNA), internal transcribed spacer (ITS) region, and mitochondrial cytochrome c oxidase subunit I (COI) gene. Predicted secondary structure models for two new species using the hypervariable region of the SSU rRNA gene and ITS2 region support the distinctness of both species. Morphological characters were subjected to principal component analysis (PCA) and genetic variations were studied in-depth to analyze the relatedness of the two new species with their congeners. An integrative approach combining morphological features, molecular analysis, and ecological characteristics was carried out to understand the phylogenetic position of the reported species within the different clades of the genus Euplotes.     

Notes on Soil Ciliates (Protozoa, Ciliophora) from The Netherlands, with Description of Keronopsis schminkei nov. spec. and Apobryophyllum schmidingeri nov. spec

Fifty-six ciliate taxa, including three new species, were found in two soil samples from the Hoge Veluwe National Park in The Netherlands. A literature search showed that The Netherlands, Belgium and Denmark are terra incognita with respect to soil ciliates: only about 100 species have been recorded. Likely, a much greater number, including many undescribed species, can be found on more detailed investigations. Two of the three new species are described in detail. Keronopsis schminkei nov. spec. differs from the congeners by the nuclear apparatus and cirral pattern. Keronopsis wetzeli?Wenzel, 1953 is neotypified with the Austrian population described by Berger and Foissner (1987). Apobryophyllum schmidingeri differs from the congeners by body shape, the macronucleus pattern (many nodules), the arrangement of the extrusomes, and various morphometric features. Scanning electron micrographs and protargol preparations of Enchelys polynucleata confirm earlier transmission electron microscopic data on the occurrence of oralized somatic monokinetids and the lack of a circumoral kinety. Extrusome length and morphology of the resting cysts are rather different in various populations of E. polynucleata, indicating high genetic diversity (subspecies?).     

Morphology of two new marine peritrich ciliates from Yellow Sea,Pseudovorticella dingi nov. spec. and P. wangi nov. spec., with supplementary descriptions of P. plicata,P. banatica and P. anomala (Ciliophora,Peritrichia)

Two new marine peritrich ciliates, Pseudovorticella dingi nov. spec. and Pseudovorticella wangi nov. spec. were collected from coastal waters of Qingdao, China. Their living morphology, infraciliature and silverline system were studied using light microscopy and silver staining methods. Pseudovorticella dingi is characterized mainly by the apically located contractile vacuole and the presence of pellicular granules. There are 28–36 and 13–18 transverse silverlines above and below the trochal band, respectively. Infundibular polykinety 3 is composed of three rows, with row 1 conspicuously shorter than the other two. P. wangi is distinguished mainly by having two ventrally located contractile vacuoles. There are 17–20 and 9–11 transverse silverlines above and below the trochal band, respectively. Abstomal end of row 1 of infundibular polykinety 3 diverges from the other two and ends alongside row 3 of infundibular polykinety 2. Pseudovorticella plicata, P. banatica, and P. anomala have been described in Sun et al. (2009); we supplement morphometric data and photographs from life and after silver staining of these three species in the present study.     

Morphological Description and Molecular Phylogeny of Two Species of Levicoleps (Ciliophora,Prostomatida), L. taehwae nov. spec. and L. biwae jejuensis nov. subspec., collected in Korea

Two colepid ciliates, Levicoleps taehwae nov. spec. and L. biwae jejuensis nov. subspec., were collected from the brackish water of the Taehwa River and a small freshwater pond in Jeju Island, South Korea, respectively. Their living morphology, infraciliature, and small subunit (SSU) rRNA gene sequences were determined using standard methods. Barrel‐shaped L. taehwae nov. spec. is a small ciliate with an average size of 45 × 25 μm in vivo, about 15 ciliary rows each composed of 12 monokinetids and two perioral dikinetids, and two 20 μm‐long caudal cilia. The sequence length and GC content of the SSU rRNA gene are 1,669 bp, 44.5%. This novel species is similar in body size to Coleps hirtus, and has six armor tiers and hirtus‐type tier plates, and the same number of ciliary rows as C. hirtus; however, it can be distinguished from the latter by the absence of armor spines and its sequence similarity of SSU rRNA gene is about 92.8% which indicates that it is a distinct form. Levicoleps biwae jejuensis nov. subspec., is a medium colepid ciliate which has a barrel‐shaped body, about 22 somatic kineties and 16 transverse ciliary rows, three mini adoral organelles, and four 15 μm‐long caudal cilia. The sequence length and GC content of the SSU rRNA gene are 1,666 bp and 44.4%.     

Spathidium seppelti foissneri nov. subspec., Spathidium simplinucleatum nov. stat., and Dileptus americanus Kahl, 1931, one new and two poorly known soil gymnostome ciliates from soils of Slovakia

The morphology and infraciliature of three soil gymnostome ciliates, Spathidium seppelti foissneri nov. subspec., Spathidium simplinucleatum nov. stat., and Dileptus americanus, collected from Slovakia, were investigated using live observation and protargol impregnation. S. seppelti foissneri is distinguished from its congeners by a combination of characters including: distinctly spatulate shape with convex oral bulge; 25 ellipsoidal, scattered macronuclear nodules; an average of 23 ciliary rows, 3 anteriorly differentiated to form a dorsal brush occupying 25% of the body length; and rod-shaped to slightly ellipsoidal extrusomes measuring about 5 x 0.5 microm. S. simplinucleatum is about 110 x 50 microm in vivo, characterized by solid, concave oral bulge, with the impression of doubled oral bulge in right side, macronuclear tortuous strand, usually 30 ciliary rows, 3 anteriorly differentiated to form a dorsal brush, and about 7 microm long rod-shaped extrusomes. S. simplinucleatum is highly variable in body size and length of monokinetidal bristle tail. Dileptus americanus is about 300 x 50 microm in vivo and differs from its congeners by a combination of characters including two macronuclear segments with a single micronucleus in between, two contractile vacuoles in dorsal side of trunk, two size types of rod-shaped extrusomes, a proboscis which occupies about 37% of body length, circular oral opening, and an average of 22 ciliary rows, of which up to 4 are differentiated to form a dorsal brush.     

Torulaspora nilssoni nov. spec.

Summary A new species of Torulaspora is described; it was isolated from some Swedish soils. This species is named Torulaspora nilssoni nov. spec. [in honour of Prof. Ragnar Nilsson (director of Institute of Microbiology, Royal Agriculture College of Sweden, Uppsala 7) and his assistant Dr. Per Eric Nilsson].     

Amphileptus aeschtae nov. spec. and Amphileptus eigneri nov. spec. (Ciliophora, Pleurostomatida), two new marine pleurostomatid ciliates from China

The morphology and infraciliature of two pleurostomatid ciliates, Amphileptus aeschtae nov. spec. and Amphileptus eigneri nov. spec., collected from shrimp-culturing ponds of the Bohai Sea, China, were investigated using live observations and the protargol impregnation method. The new species A. aeschtae is identified by the presence of 200-300 macronuclear nodules and several contractile vacuoles along the ventral side of the cell. A. eigneri differs from its congeners, inter alia, by the marine habitat, the presence of many contractile vacuoles along the ventral margin and the number of somatic kineties on both sides. Loxophyllum ozakii Shigematsu, 1953 is transferred to Kentrophyllum.     

Re-establishment of the cyrtophorid genus Coeloperix Deroux,nov. gen., with a description of Coeloperix sleighi nov. spec. (Protozoa,Ciliophora, Cyrtophorida)

The cyrtophorid genus Coeloperix, which has remained invalid for over 20 years, is re-established. This taxon is characterized by Lynchellids without podites on the ventral side; somatic kineties making no noticeable naked gap between left and right areas; pre- and post-oral kineties completely separated; two terminal fragments; perioral kineties consisting of one continuous anterior and two detached posterior rows; with cross-striated band around perimeter between ventral and dorsal surfaces. Based on this definition, two nominal species formerly placed in the invalid genus have been included again in Coeloperix: Coeloperix dirempta (Deroux, 1970) nov. comb. [synonym: Lynchella dirempta; Deroux, 1970], C. aspidisciformis (Kahl, 1933) nov. comb. [syn. Lynchella aspidisciformis; Kahl, 1933]. Another two morphotypes C. eforiana (Tucolesco, 1962) nov. comb. [syn. Lynchella eforiana; Tucolesco, 1962] and C. lynchelliformis (Borror, 1972) nov. comb. [syn. Chlamydodon lynchelliformis; Borro, 1972] have been transferred into the new genus. The morphology of living cells and infraciliature of a new species, C. sleighi nov. spec., isolated from the coastal area of China, have been investigated. It is diagnosed by: size about 40×30 μm in vivo; consistently 4 preoral and 15–16 postoral kineties; 12–16 nematodesmata; cross-striated band with two separations in the equatorial area; 3–4 (usually 3) finger-like tentacles on the ventral side; macronucleus ellipsoidal; two contractile vacuoles diagonally located; marine habitat.     

Two new species of cossonine weevils,Araucarius kuscheli spec. nov. and A. crassipunctatus spec. nov. (Coleoptera: Curculionidae) from southern Brazil

The occurrence of cossonine weevils of the genus Araucarius Kuschel 1966 is limited to South American Araucaria trees. Adults and larvae of two new species, Araucarius kuscheli spec. nov. and A. crassipunctatus spec. nov. have been found under the bark of Araucaria angustifolia trees in São Francisco de Paula, Rio Grande do Sul, Brazil. A key for the adults of the genus Araucarius including the two new species and the already known two Brazilian and four Chilean species is presented.     

Rigidothrix goiseri nov. gen., nov. spec. (Rigidotrichidae nov. fam.), a new "flagship" ciliate from the Niger floodplain breaks the flexibility-dogma in the classification of stichotrichine spirotrichs (Ciliophora, Spirotrichea)

Rigidothrix goiseri nov. gen., nov. spec. was discovered in soil from the Niger floodplain near to the town of Timbuktu, Republic of Mali. Its morphology, ontogenesis, and 18S rDNA gene sequence were studied with standard methods. Rigidothrix goiseri is very conspicuous in vivo because of its average size of 230x70 microm and a distinct tail. Further main characteristics include the rigid body; the conspicuous, stylonychid frontal area; the undulating membranes in Oxytricha pattern; a mighty adoral zone of membranelles not reorganized during ontogenesis; distinct midventral rows of which those of the opisthe develop right of the parental ones; and eight dorsal kineties, of which three develop intrakinetally and five are generated dorsomarginally. Rigidothrix goiseri possesses main characteristics of oxytrichine, stylonychine, and urostyline stichotrichs, specifically, it is the first rigid stichotrich with midventral rows, and thus it breaks the flexibility dogma. Distinct similarities with the genus Uroleptus and the CEUU hypothesis suggest that R. goiseri is more closely related to the oxytrichine than urostyline stichotrichs, in spite of the conspicuous midventral rows. This is also supported by the sequence data which show that R. goiseri is almost equally similar to both Oxytricha granulifera (95.88%) and Uroleptus gallina (94.93%), but fairly different from Urostyla grandis (92.7%). The main morphological peculiarities of R. goiseri show that it represents a new genus which should be classified into a new family, the Rigidotrichidae, together with the genera Uroleptus, Territricha, and Afrophrya nov. gen., the latter comprising Rigidothrix-like stichotrichs with cyrtohymenid oral apparatus. The conspicuous size and shape make R. goiseri a biogeographic flagship likely confined to Africa.