Morphological and phylogenetical studies on a new soil hypotrich ciliate: Kahliella matisi spec. nov. (Hypotrichia,Kahliellidae)

The morphology, ontogenesis, encystment, and 18S rRNA gene sequence of a new soil hypotrich ciliate, Kahliella matisi, were studied. Main characteristics of K. matisi are: (1) two short and six longitudinal cirral rows right of the adoral zone of membranelles and four longitudinal rows left of it; (2) three dorsal kineties, of which kinety 1 extends along the left cell margin, kinety 2 runs in a slightly sigmoidal line, and kinety 3 is distinctly shortened posteriorly. Ontogenesis is similar to that in congeners, especially in the development of the marginal rows and long dorsal kineties, the preservation of some old cirral rows after division, and the direction of the neokinetal wave. However, there are some peculiarities: (1) reorganization of the proximal parental adoral membranelles; (2) splitting of opisthe's anlage II into the cirral streak II and III; and (3) formation of the parental cirral row R3 from anlagen IV and V. During encystment, the body diminishes and becomes globular, the nuclear apparatus is reorganized, and the ciliature is resorbed. In our molecular phylogenies, the family Kahliellidae is polyphyletic and the position of K. matisi is rather poorly resolved, indicating a relationship with oxytrichids.     

Morphology and morphogenesis of a new marine cyrtophorid ciliate, Hartmannula sinica nov. spec. (Protozoa, Ciliophora, Cyrtophorida) from China

The living morphology, infraciliature and morphogenesis of a new marine cyrtophorid ciliate, Hartmannula sinica nov. spec., collected from Qingdao, north China, have been investigated. The new species is characterized by: size in vivo 90–130×40–50 μm, body long elliptical in outline, cilium-free field covered with a conspicuous alveolar layer; 24–31 ventral kineties, the rightmost 6–9 of which extend apically; 20–24 nematodesmal rods; about 15 sparsely distributed contractile vacuoles; a yellowish pigment spot is always present near the anterior tip of the cell. Morphogenetic events exhibit a pattern, which is homologous with that of related cyrtophorids. The main features are as follows: (1) preoral and circumoral kineties of opisthe develop from the oral primordium that forms in mid-body from sections of 3 postoral kineties, while parental preoral and circumoral kineties are retained; (2) parental cytostome and nematodesmal rods are resorbed in middle divisional stages and then replaced by newly formed structures; (3) the heteromerous macronucleus unifies in the late divisional stage.     

Observations on soil haptorid ciliates (Protozoa,Ciliophora) from Slovakia

The morphology and morphometry of five soil haptorid ciliates (Apospathidium terricola, Diplites telmatobius, Armatoenchelys geleii, Paraenchelys terricola, and Spathidium aciculare), collected from a variety of terrestrial habitats in Slovakia (Central Europe), were investigated using live observation and protargol impregnation. Emphasis was given on the ciliary pattern, the fine structure of dorsal brush, as well as the shape and size of the extrusomes. The Slovak specimens of A. geleii differ considerably from other populations investigated in detail by body size and micronuclei number. Furthermore, the most important difference concerns the number of dorsal brush rows (4 vs. 3). Therefore, it was decided to establish a new subspecies, namely Armatoenchelys geleii barborae ssp. n., for the Slovak population. Shape and nuclear variants of A. terricola and P. terricola are presented and morphometrically characterized. Novel details on the dorsal brush structure of A. terricola and D. telmatobius are reported. Pair formation and exconjugant nuclear reconstruction in P. terricola are described. Two species, namely A. terricola and S. aciculare, are first records for the Holoarctic biogeographic region.     

Morphology,ontogenesis and molecular phylogeny of a new saline soil ciliate,Uroleptoides salina nov. spec. (Ciliophora,Hypotrichia)

The morphology, morphogenesis and molecular phylogeny of a new saline soil hypotrich ciliate, Uroleptoides salina nov. spec., discovered from China, was investigated. The new species is characterized as follows: body 150–215 × 40–50 μm in vivo, slender and highly flexible; usually four ellipsoidal macronuclear nodules; contractile vacuole absent; cortical granules absent; endosymbiotic algae present; amphisiellid median cirral row consists of 14–25 cirri and terminates about 47% down length of body; usually three buccal cirri and 3–13 cirri left of anterior portion of amphisiellid median cirral row; 3–5 transverse cirri. Morphogenesis during binary fission is characterized by: (1) the parental adoral zone of membranelles is retained completely, parental paroral contributes to the formation of the undulating membranes anlage for the proter; (2) the oral primordium of the opisthe is formed apokinetally; and (3) the amphisiellid median cirral row is formed from two anlagen. Phylogenetic analyses based on SSU rDNA sequence data show that Uroleptoides salina nov. spec. has a close relationship with its morphologically similar species, U. longiseries, U. magnigranulosus, Orthamphisiella breviseries, and Parabistichella variabilis.     

Studies on the ectocommensal ciliate,Trachelostyla tani nov. spec. (Protozoa: Ciliophora: Hypotrichida) from the mantle cavity of the scallop Chlamys farrei

A total of 33 sites (27 temporary pools, five dune slacks and one shallow lake) were sampled during January 1998. Forty two taxa (one ctenopoda, 19 anomopoda and 22 copepoda) are recorded. Data from the varied habitats types (temporary pools, dunary ponds, shallow lakes) are compared and biotic and abiotic factors influencing the spatial and temporal distribution of local zooplankton are discussed. The paper concludes with comments why local conservation efforts need to be directed towards temporary pools and dune slacks, two much-neglected habitats.     

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 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”.     

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).     

Schmidingerothrix extraordinaria nov. gen., nov. spec., a secondarily oligomerized hypotrich (Ciliophora, Hypotricha, Schmidingerotrichidae nov. fam.) from hypersaline soils of Africa

Schmidingerothrix extraordinaria nov. gen., nov. spec. was discovered in hypersaline soils from Namibia and Egypt. Its morphology and ontogenesis were studied with standard methods. Schmidingerothrix extraordinaria is a highly flexible, slender hypotrich with an average size of 90 × 15 μm. Likely, it prefers a salinity around 100‰ and feeds mainly on bacteria. Schmidingerothrix is extraordinary in having a frayed buccal lip, three-rowed adoral membranelles, only one frontal cirrus, a distinct gap between frontal and ventral adoral membranelles, and a miniaturized first frontal membranelle, while a paroral membrane, dorsal bristle rows, and buccal, transverse, and caudal cirri are absent. The ontogenesis is simple: the opisthe oral apparatus and frontoventral ciliature originate de novo, while parental structures are involved in the development of the ciliature of the proter. This special organization is used to define a new family, the Schmidingerotrichidae, which is likely related to the Cladotrichidae. Schmidingerothrix extraordinaria is very likely a secondarily oligomerized hypotrich, and the reduction occurred possibly very long ago because no traces of the ancestral ciliature remained in the ontogenetic processes. Possibly, the simple ciliature is an adaptation to highly saline habitats, where competition is low and bacterial food abundant.     

Morphology and molecular phylogeny of the anaerobic freshwater ciliate Urostomides spinosus nov. spec. (Ciliophora,Armophorea, Metopida) from China

The morphology and molecular phylogeny of a new metopid ciliate, Urostomides spinosus nov. spec., discovered in a freshwater ditch in Qingdao, China, were investigated using live observation, morphometry and protargol staining as well as molecular phylogenetic methods. Diagnostic features of the new species include a broadly obpyriform body carrying three posterior spines, eight somatic kineties, five preoral dome kineties with specialized row 3, adoral zone composed of about 28 membranelles, making a 270° turn around body axis. Phylogenetic analyses of the SSU rDNA sequence revealed that the genus Urostomides is monophyletic, but its interspecific relationships remained unresolved. Moreover, a closer relationship of the new species with the morphologically similar Urostomides campanula was not supported by the molecular data.     

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.     

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.     

A huge,undescribed soil ciliate (Protozoa: Ciliophora) diversity in natural forest stands of Central Europe

We investigated 12 natural forest stands in eastern Austria for soil ciliate diversity, viz., eight beech forests and two lowland and Pinus nigra forests each. The stands span a wide range of climatic (e.g., 543–1759 mm precipitation, 160–1035 m above sea-level) and abiotic (e.g., pH 4–7.4) factors. Samples were taken twice in autumn and late spring and analysed with the non-flooded Petri dish method. Species were identified in vivo, in silver preparations, and in the scanning electron microscope. A total of 233 species were found, of which 30 were undescribed, a surprising number showing our ignorance of soil ciliate diversity, even in Central Europe. Species number varied highly from 45 (acidic beech on silicate) to 120 (floodplain forest) and was strongly correlated with pH and overall habitat quality, as measured by climate, the C/P quotient (ratio of r-selected colpodean and k-selected polyhymenophorean ciliates), and the proportion of mycophagous ciliate species; multivariate analysis showed further important variables, viz., the general nutrient status (glucose, nitrogen, C/ N ratio) and microbial (urease) activity. The highest species number occurred in one of the two floodplain soils, supporting the intermediate disturbance hypothesis. The three main forest types could be clearly distinguished by their ciliate communities, using similarity indices and multidimensional scaling. Individual numbers varied highly from 135^–1 (lowland forest) to 10,925 ml^–1 (beech on silicate) soil percolate and showed, interestingly, a weak correlation with soil protozoan phospholipid fatty acids. Eight of the 30 new species found and a forgotten species, Arcuospathidium coemeterii (Kahl 1943) nov. comb., are described in detail, as examples of how species were recognized and soil protozoan diversity should be analyzed: Latispathidium truncatum bimicronucleatum, Protospathidium fusioplites, Erimophrya sylvatica, E. quadrinucleata, Paragonostomum simplex, Periholosticha paucicirrata, P. sylvatica, and Australocirrus zechmeisterae.     

Morphology and morphogenesis of a new marine ciliate, Apokeronopsis bergeri nov. spec. (Ciliophora, Hypotrichida), from the Yellow Sea, China

The morphology and morphogenesis of a new marine hypotrich ciliate, Apokeronopsis bergeri nov. spec., collected from mussel-farming waters near Qingdao, China, are described from living and protargol-impregnated specimens. This ciliate has characteristics that place it in the family Pseudokeronopsidae, namely, two long rows of frontal cirri (bicorona), which are continuous with the long midventral rows, and a single row of marginal cirri on each side of the body. It shares with its only congener, Apokeronopsis crassa, the long rows of buccal and transverse cirri and the wide spacing between the midventral rows of cirri. These characters separate the genus Apokeronopsis from Pseudokeronopsis, which has a single buccal cirrus, fewer transverse cirri and midventral rows of cirri arranged in a typical zig-zag pattern. A. bergeri differs from A. crassa in its shape, colour and in the numbers of membranelles and transverse cirri. Although morphogenesis in A. bergeri is similar to that of A. crassa in most respects, the mode of formation of the buccal cirri is slightly different. The close relationship of A. bergeri with A. crassa, and the more distant relationship with three Pseudokeronopsis species, is supported by a comparison of the sequences of their ITS1-5.8S-ITS2 rDNA regions.     

Revision of the genus Stentor Oken (Protozoa, Ciliophora) and description of S.araucanus nov. spec, from South American lakes

Stentor is a heterolrich ciliate which often forms lawn-likecovers on the bottom and/or blooms in the pelagial of lakesworldwide. The species involved in these spectacular eventswere usually either not determined or misidentified becausethe keys are outdated and incomplete. Thus, we have revisedthe nominal species described since the first major revisionby Ehrenberg (1838). Main species characteristics are the presence/absenceof symbiotic algae, the shape of the macronucleus and the colourof the cortical pigment granules. The last character mentionedmust be studied in live cells because the pigment bleaches inchemically fixed specimens. Nineteen valid species are recognizedand dichotomously keyed according to these characteristics.Twenty-seven other species and varieties, described after Ehrenberg'srevision, are synonyms or species indeterminata A new species.S.araucanus, is described from South American lakes. It is asmall, broadly trumpet-shaped Stentor with symbiotic algae,vermiform macronucleus and blue-green cortical granules. Stentoraraucanus is probably euplanktic and restricted to the southernhemisphere. Stentor auriculalus Kahl. 1932 sensu Wang (1934)is recognized as a new species, Condylostoma wangi, and transferredto the genus Condylostoma. New nomenclatural corrections: Stentorbaicalius nom. nov. (pro S.pygmaeus, preoccupied). S loricatiisnom. corr. (for S.loricata), S.ruber nom. corr. (for S.ruhra).     

Cotterillia bromelicola nov. gen., nov. spec., a gonostomatid ciliate (Ciliophora, Hypotricha) from tank bromeliads (Bromeliaceae) with de novo originating dorsal kineties

Cotterillia bromelicola nov. gen., nov. spec. was discovered in the tanks of the Mexican bromeliad Tillandsia heterophylla. Its morphology, ontogenesis, and 18S rDNA were studied with standard methods. Cotterillia has many cirral rows on both sides of the body. Uniquely, and thus used to diagnose the new genus Cotterillia, it has dorsal kineties originating de novo, producing neokinetal waves where the parental dorsal kineties reorganize to “combined rows”, consisting of dorsal bristles anteriorly and of cirri posteriorly. Thus, up to four generations of bristles and cirri occur on the dorsal body surface. Cotterillia bromelicola has a gonostomatid body and adoral zone of membranelles, while the dense ciliature and the neokinetal waves resemble kahliellid hypotrichs. However, the de novo origin of anlage 1 and the molecular analyses show convincingly that Cotterillia belongs to the GonostomatidaeSmall and Lynn, 1985, for which an improved diagnosis is provided. Thus, neokinetal waves originated several times independently. The molecular differences between Trachelostyla, Gonostomum, and Cotterillia are small (≤5%) compared to their distinct morphologies and ontogeneses, suggesting that the 18S rDNA underestimates generic diversity. Our study emphasizes the need of combined morphological, ontogenetic, and molecular investigations to unravel the complex phylogeny and evolution of hypotrich ciliates.