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.     

Novistrombidium sinicum n. sp. and Novistrombidium orientale n. sp. (Protozoa: Ciliophora): Two New Oligotrich Ciliates from a Mangrove Wetland, South China

ABSTRACT. The morphology and infraciliature of two new marine oligotrich ciliates, Novistrombidium sinicum n. sp. and Novistrombidium orientale n. sp., isolated from a mangrove wetland near Guangzhou, southern China, were studied from live and protargol-stained specimens. Novistrombidium sinicum is different from its congeners by the combination of the following characters: three posteriorly directed thigmotactic membranelles, one ellipsoidal macronucleus, the extrusomes equidistantly arranged, and the ventral kinety commencing below the right end of the girdle kinety. The small-sized N. orientale can be separated from its congeners by two posteriorly directed thigmotactic membranelles, one ellipsoidal macronucleus, the extrusomes equidistantly arranged, and the ventral kinety commencing below the right end of the girdle kinety.     

Two vicariant Semispathidium species from tropical Africa and central Europe: S. fraterculum nov. spec. and S. pulchrum nov. spec. (Ciliophora,Haptorida)

Using standard methods, we describe two new Semispathidium species from semiterrestrial habitats of tropical Africa and central Europe. Semispathidium fraterculum nov. spec. and S. pulchrum nov. spec. differ from each other and from two Namibian (Southwest African) congeners by body size, the shape and location of the extrusomes; the shape of the macronucleus, the number of ciliary rows, the number of cilia within the rows, and the number of dikinetids comprising the dorsal brush rows. The globular resting cyst of S. fraterculum is unique in having countless granules on the inside of the external layer, forming sharp-angled rows. The distribution of these and two further, not yet described south African species indicates that the genus Semispathidium evolved in subtropical Africa. Semispathidium fraterculum and S. pulchrum are highly similar, both morphologically and ecologically, differing in mainly one important feature: the shape and location of the extrusomes used for prey capture. This suggests that they evolved from a common ancestor whose areal was divided by a vicariant event, causing an independent evolution of the split populations. We argue that this and other species pairs indicate that the vicariance speciation model is applicable to protists.     

Morphology and morphometry of Bresslauides pratensis n. sp., a new soil ciliate (Ciliophora, Colpodea) from Turkey

A new colpodid ciliate, Bresslauides pratensis n. sp., was discovered in soil from a meadow field in Turkey. Its morphology was investigated using live observation, protargol impregnation, silver nitrate impregnation, and scanning electron microscopy. The new species differs from the congeners by the following combination of features: the cell size in vivo is about 135-180 μm × 130-160 μm; the body has an almost circular outline with a conspicuous bulge on the ventral side; the somatic cilia are arranged in about 105 densely spaced, sigmoidal kineties; a diagonal groove extends to the left cell side, terminating in a pronounced postoral sack; one almost globular macronucleus and usually two globular micronuclei; the oral structures occupy almost the anterior half of the cell and comprise a distally elongated right polykinetid and a crescentic left oral polykinetid, both restricted to the vestibulum; on average 12 vestibular, 31 postoral, and 55 left oral kineties. Based on the morphological data, it was concluded that B. pratensis is a well-outlined and distinctive member of the genus Bresslauides. Additionally, the most recent molecular data on the order Colpodida and genera Colpoda, Bresslaua, and Bresslauides are briefly discussed.     

Morphology,Conjugation, and Postconjugational Reorganization of Dileptus tirjakovae n. sp. (Ciliophora,Haptoria)

ABSTRACT. We studied the morphology, conjugation, and postconjugational reorganization of a new haptorid ciliate, Dileptus tirjakovae n. sp., using conventional methods. Dileptus tirjakovae is characterized by two abutting, globular macronuclear nodules and scattered brush kinetids. Conjugation is similar to that in congeners, that is, it is temporary, heteropolar, and the partners unite bulge‐to‐bulge with the proboscis. Some peculiarities occur in the nuclear processes: there are two synkaryon divisions producing four synkaryon derivatives, of which two become macronuclear anlagen, one becomes the micronucleus, and one degenerates. Unlike spathidiids, D. tirjakovae shows massive changes in body shape and ciliary pattern before, during, and after conjugation: early and late conjugants as well as early exconjugants resemble Spathidium, while mid‐conjugants resemble Enchelyodon. These data give support to the hypothesis that spathidiids evolved from a Dileptus‐like ancestor by reduction of the proboscis. Dileptus tirjakovae exconjugants differ from vegetative cells by their smaller size, stouter body, shorter proboscis, and by the lower number of ciliary rows, suggesting one or several postconjugation divisions. Although 83% of the exconjugants have the vegetative nuclear pattern, some strongly deviating specimens occur and might be mistaken for distinct species, especially because exconjugants are less than half as long as vegetative cells.     

Description of Deviata rositae n. sp., a new ciliate species (Ciliophora, Stichotrichia) from Argentina

Soil samples were taken from a temporary pond located in Buenos Aires province, Argentina, during the dry phase in the summer of 2005. The ciliates were studied alive and after staining with protargol. Deviata rositae n. sp. measures 112-154 microm in length and 21-28 microm in width in vivo and has a vermiform body. The contractile vacuole is located in the mid-body on the left. The macronucleus is moniliform and there are 1-3 micronuclei. The oral apparatus is composed of 14-18 adoral membranelles and straight paroral and endoral membranes that never intersect each other. The somatic ciliature is arranged in four frontal cirri, one buccal cirrus, six long and slightly spiraled rows of cirri with the first right row extending up to the equatorial or sub-equatorial region, and two dorsal rows of dikinetids. This new species of Deviata primarily differs from its congeners by the number of macronuclear nodules and the number and disposition of the dorsal rows of dikinetids.     

Urotricha psenneri n. sp. and Amphileptus piger (Vuxanovici, 1962) n. comb., two planktonic ciliates (Protozoa, Ciliophora) from an oligotrophic lake in Austria

Two euplanktonic ciliates, Urotricha psenneri n. sp. (Prostomatida) and Amphileptus piger (Vuxanovici, 1962) n. comb. (Pleurostomatida), were discovered in the surface plankton of the oligotrophic Lake Traunsee in Austria. Their morphology and infraciliature were studied in live cells as well as in specimens impregnated with protargol and silver nitrate. Urotricha psenneri is a small urotrichid, less than 50 microm length and with a single caudal cilium. It is unique in having (i) a massive oral basket projecting as a conspicuous bulge with cylindrical microfibrillar annulus and (ii) a curved brosse row 1 in the broad, barren circumoral area. Amphileptus piger (Vuxanovici, 1962) is about 55 x 13 microm in vivo, has two macronuclear nodules with a single micronucleus in between in the posterior body half, has a single contractile vacuole with a terminal excretory pore, and few, but thick and thus highly conspicuous extrusomes. The amphileptid ciliary pattern (spica) is difficult to recognise due to the widely spaced basal bodies.     

Morphology and ciliary pattern of some rare haptorid ciliates, with a description of the new family Kamburophryidae (Protists, Haptoria)

We investigated three rare haptorid ciliates, viz., Lagynophrya gibba Kahl (1935), Enchelys lajacola nov. spec., and Spathidium implicatum Kahl (1930), using live observation, silver impregnation, morphometry, and scanning electron microscopy. Lagynophrya gibba, which was rediscovered in peatland soil from Iceland, is referred to a new genus, Kamburophrys, and a new family, Kamburophryidae, based on a unique organelle, the brush membranoid. This structure is near the dorsal brush and composed of very narrowly spaced cilia, about 5 μm long. The genus Kamburophrys has a unique combination of features, viz., an oral cone on the oral bulge, an oblique circumoral kinety, and a subapical hump carrying the three-rowed dorsal brush and the brush membranoid. The Kamburophryidae possibly belong to the order Spathidiida. Enchelys lajacola was discovered in mud from granitic rock-pools (Lajas) in Venezuela, South America. The new species is characterized by a bottle-like shape, a macronucleus with the shape of a curved strand, a heterostichad dorsal brush, and rod-shaped toxicysts. Spathidium implicatum, which was rediscovered in an ephemeral meadow puddle near Salzburg city centre, is neotypified and referred to the genus Apertospathula because it has an open circumoral kinety.     

Morphology and infraciliature of three species of Metaurostylopsis (Ciliophora, Stichotrichia): M. songi n. sp., M. salina n. sp., and M. marina (Kahl 1932) from sediments, saline ponds, and coastal waters

Two new urostylid ciliates, Metaurostylopsis songi n. sp. and Metaurostylopsis salina n. sp. and Metaurostylopsis marina (Kahl 1932) are investigated using live observation and protargol impregnation. These species were isolated in Korea from intertidal sediments, saline ponds, and coastal waters. Metaurostylopsis songi is in vivo about 120 microm x 25 microm, has a slenderly ellipsoidal body, colorless cortical granules in rows on ventral and dorsal body sides, about 54 macronuclear nodules, 28-47 adoral membranelles, five frontal, two or three frontoterminal and six or seven transverse cirri, and 9-12 midventral cirral pairs followed posteriorly by 1-3 single cirri. In vivo M. salina is about 60 microm x 25 microm, has a pyriform body, colorless cortical granules irregularly arranged, about 45 macronuclear nodules, 18-23 adoral membranelles, three frontal, three to five frontoterminal and two to five transverse cirri, and four or five midventral cirral pairs followed posteriorly by five to seven single cirri. Both species have three marginal cirral rows on each body side and 3 long dorsal kineties. The Korean specimens of M. marina match the Chinese population in all main features. Metaurostylopsis songi differs from M. marina by the more slender body, the number of frontal cirri (invariably five vs. four), and the arrangement of cortical granules (in rows on dorsal and ventral cortex vs. only along dorsal kineties and anterior body margin). Metaurostylopsis salina differs from its congeners by the distinctly smaller size, the pyriform body shape, the scattered cortical granules (vs. in rows), and number of frontal cirri. It differs from M. marina also by the number of midventral cirral pairs (four or five vs. seven to 11).     

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?).     

Dexiotrichides pangi n. sp. (protozoa,ciliophora, scuticociliatia), a new marine ciliate from the north China sea

The morphology, infraciliature, and silverline system of a new marine scuticociliate, Dexiotrichides pangi n. sp. were investigated. The new species is characterized by: size about 45-65 x 20-25 microm in vivo with kidney-like body shape and obliquely truncated semicircle-shaped apical plate; cytostome at bottom of conspicuously depressed oral cavity, which is located at the cell equatorial level; paroral membrane extending anteriorly to membranelle 3; scutica multi-rowed; 33-38 somatic kineties; contractile vacuole near ventral side and subcaudally positioned, opening at posterior end of somatic kinety 3; one oval macronucleus and one small micronucleus; caudal cilium positioned in a small pouch; marine habitat. Based on the data obtained, an improved diagnosis for the genus Dexiotrichides is suggested: body with circular cross-section and conspicuous cilia-free apical plate; buccal cavity conspicuously depressed with cytostome located near or at equatorial level; three membranelles transversely orientated each with 2-3 rows; paroral membrane zigzaging structure, extending to about half of the length of buccal field; multi-rowed scutica; somatic kinety one strongly shortened and terminating anteriorly at posterior end of buccal field; basal bodies in equatorial region arranged usually in circular pattern, while in the anterior portion of somatic kinety 2, basal bodies characteristically in pairs and separated from the posterior part of kinety 2; one caudal cilium.     

Morphologic and molecular characterization of Apertospathula pilata n. sp., a novel freshwater spathidiid (Ciliophora,Litostomatea) from Idaho,USA

Order Spathidiida Foissner and Foissner, 1988 comprises a large group of morphologically diverse, primarily predatory, free living ciliates, the phylogeny of which has remained stubbornly unresolved. Families Arcuospathidiidae and Apertospathulidae are two morphologically similar groups established on the basis of differences in the morphology of the oral bulge and circumoral kinety. While Arcuospathidiidae is non-monophyletic in 18S rRNA gene analyses, the Apertospathulidae has been represented by only a single Apertospathula sequence in public databases. In this report, a novel freshwater species, Apertospathula pilata n. sp. is described on the basis of living observation, silver impregnation, and scanning electron microscopy. The phylogeny of the new species is assessed based on the rRNA cistron. The main features distinguishing A. pilata n. sp. from all congeners are: the oral bulge extrusomes (filiform, up to 25 µm long), the combination of body size (130–193 µm) and shape (spatulate), the extensive oral bulge length (41% of the cell length after protargol impregnation), and multiple micronuclei (one to five, two on average). The monophyly of Apertospathulidae Foissner, Xu and Kreutz, 2005 is rejected.     

Kiitricha minuta n. sp., a peculiar hypotrichous ciliate (Ciliophora, Spirotrichea) from the Yellow Sea

The morphology and infraciliature of a new ciliate, Kiitricha minuta n. sp., isolated from the Yellow Sea, were investigated using live observation and protargol impregnation. Kiitricha minuta represents a third member of the rarely known order Kiitrichida. It is unique in the subclass Hypotrichia in having many rows of small uniform cirri along the right side of the body and the dorsal kineties composed of dikinetids, most of which bear two cilia. Kiitricha minuta n. sp. is ovoid and measures about 60 x 45 microm in vivo. It has a huge buccal cavity occupying about 80% of the body length, numerous body extrusomes, one macronucleus and two micronuclei, 27-27 adoral membranelles, 9-12 frontoventral cirral rows, a submarginal row of 7-9 cirri, 6 or 7 transverse cirri, and roughly 7-9 dorsal kineties. This new species differs distinctly from its only congener Kiitricha marina by its smaller size (60 microm vs. 80-150 microm), the presence of body extrusomes (vs. absent), the different macronuclear pattern (one vs. two nodules), and the lower number of frontoventral cirral rows (9-12 vs. 21-26), which terminate at the anterior two-thirds of body (vs. extend to the posterior). The new term "submarginal cirral row" is introduced to distinguish from the marginal cirral row in typical hypotrichs sensu lato. Based on our new observations and the literature, an improved diagnosis for the genus Kiitricha is provided and its phylogenetic importance is discussed.     

A new oligotrich (Ciliophora,Oligotrichia) from Argentina,with redefinition of Novistrombidium Song and Bradbury

A new oligotrich similar to Novistrombidium was discovered in plankton samples from an artificial tributary of the Salado River, Buenos Aires province, Argentina, in summer 2010. Propecingulum fistoleramalliei sp. n. has an obovate and anteriorly truncated body, with a conspicuous ventral furrow, is flattened ventrally, and has a prominent right apical protrusion. It temporarily attaches to the substratum by a posterior mucous thread. Rod-shaped extrusomes arranged equidistantly and insert directly above the girdle kinety. The macronucleus is globular to ellipsoidal. The contractile vacuole is located in the left, anterior quarter of the cell and the adoral zone is composed of 30–35 collar, 9–14 buccal, and two thigmotactic membranelles. The girdle kinety is dextrally spiraled and ventrally open; the ventral kinety is posterior to anterior end of the girdle kinety. The oral primordium develops posterior to the right thigmotactic membranelle and anterior the stripe of extrusomes above left, lateral portion of the girdle kinety. The SSUrDNA phylogeny confirms one more time that Novistrombidium is not monophyletic; consequently, we elevate the subgenus Propecingulum up to genus rank and redefine the genus Novistrombidium.     

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.     

Description of Four New Soil Dileptids (Ciliophora, Haptoria), with Notes on Adaptations to the Soil Environment

We studied the morphology of four new dileptid ciliates, using standard methods. Dileptus microstoma, which was discovered in Benin (Africa), is outstanding in having a very small oral opening (~4 μm), an interrupted dorsal row of contractile vacuoles, and ampulliform extrusomes. Dileptus semiarmatus, which was discovered in Austria, possesses extrusomes only in the right posterior half of the proboscis and has very widely spaced circumoral and perioral kinetids. Dileptus longitrichus, which was discovered in Japan, is almost unique in having up to 15 μm long brush bristles and a row of contractile vacuoles each in ventral and dorsal side of body. Pseudomonilicaryon brachyproboscis, which was discovered in Greece, differs from the congeners by the narrowly ellipsoidal micronuclei, the dimorphic dorsal brush, the extrusomes, and the contractile vacuole pattern. Four new features are introduced for distinguishing species in dileptids: shape of micronucleus, monomorphic/dimorphic dorsal brush, shape of oral opening, and spacing of circumoral dikinetids. The terrestrial dileptids share several distinct morphological features that are probably adaptations to the soil environment: (1) the body is comparatively slender and small, what is likely related to the narrowness of the habitat; (2) the proboscis is short, which increases the relative volume of the trunk, what might be related to its fragility and/or to the space available for prey digestion; (3) the long dorsal bristles might foster prey recognition; and (4) the pronounced body flexibility in all dileptids likely fosters their high diversity in the narrow and wrinkled soil habitat.     

Morphological and molecular characterization of the name-bearing type species Rimaleptus binucleatus (Kahl, 1931), with a phylogenetic re-analysis of dileptid evolutionary history (Ciliophora: Litostomatea: Rhynchostomatia)

We collected Rimaleptus binucleatus from soil in the surroundings of the town of Ulsan, South Korea. Its morphology and 18S rRNA gene were studied using standard methods. This possibly widely distributed species is characterized by: (i) a size of about 170–400 × 20–65 μm; (ii) a narrowly to cylindrically dileptid body with proboscis occupying about 30–65% of body length; (iii) two dorsal contractile vacuoles; (iv) two size groups of rod-shaped extrusomes; and (v) about 18–29 ciliary rows, 4–6 of them anteriorly differentiated into a staggered dorsal brush. Phylogenetic analyses of five new rhynchostomatian 18S rRNA gene sequences supported monophylies of the orders Tracheliida and Dileptida, but revealed that the genera Rimaleptus and Pseudomonilicaryon are polyphyletic. Monophyly of genera with two macronuclear nodules was consistently rejected, but monophylies of dileptids with many scattered macronuclear nodules and of dileptids with moniliform macronucleus and multi-rowed dorsal brush could not be excluded by statistical topology tests. Nevertheless, phylogenetic network analyses indicated considerable conflict in the phylogenetic signal provided by the 18S rRNA gene to resolve unambiguously relationships among dileptid genera.     

Nematodes from Swainson's spurfowl Pternistis swainsonii and an Orange River francolin Scleroptila levaillantoides in Free State Province, South Africa, with a description of Tetrameres swainsonii n. sp. (Nematoda: Tetrameridae)

Five Swainson's spurfowl collected in Free State Province, South Africa, were examined for helminth parasites, and the nematodes Acuaria gruveli, Cyrnea parroti, Gongylonema congolense, Subulura dentigera, Subulura suctoria and a new Tetrameres species were recovered. Their respective prevalence was 100, 20, 80, 20, 20 and 20%. These nematodes are all new parasite records for Swainson's spurfowl, and Acuaria gruveli constitutes a new geographical record as well. A single specimen of Cyrnea eurycerca was found in an Orange River francolin, representing a new host and geographical record for this parasite. The new species, for which the name Tetrameres swainsonii is proposed, can be differentiated from its congeners by a combination of the following characters of males: two rows of body spines, a single spicule which is 1152-1392 microm long, and eight pairs of caudal spines arranged in two ventral and two lateral rows of four spines each. The single female has the globular shape typical of the genus.     

Ontogenesis of Dileptus terrenus and Pseudomonilicaryon brachyproboscis (Ciliophora, Haptoria)

ABSTRACT. Dileptids are haptorid ciliates with a conspicuous proboscis belonging to the oral apparatus and carrying a complex, unique ciliary pattern. We studied development of body shape, ciliary pattern, and nuclear apparatus during and after binary fission of Dileptus terrenus using protargol impregnation. Additional data were obtained from a related species, Pseudomonilicaryon brachyproboscis . Division is homothetogenic and occurs in freely motile condition. The macronucleus is homomeric and condenses to a globular mass in mid-dividers. The proboscis appears in late mid-dividers as a small convexity in the opisthe's dorsal brush area and maturates post-divisionally. The oral and dorsal brush structures develop by three rounds of basal body proliferation. The first round generates minute anarchic fields that will become circumoral kinetofragments, while the second round produces the perioral kinety on the right and the preoral kineties on the dorsal opisthe's side. The dorsal brush is formed later by a third round of basal body production. The formation of various Spathidium -like body shapes and ciliary patterns during ontogenesis and conjugation of Dileptus shows a close relationship between spathidiids and dileptids. On the other hand, the peculiarities of the dileptid morphology and ontogenesis indicate a long, independent evolution.