Morphological Characterizations of Four Species of Parallelostrombidium (Ciliophora,Oligotrichia), with a Note on the Phylogeny of the Genus

The morphology and phylogeny of four oligotrichid ciliates, Parallelostrombidium paraellipticum sp. n., P. dragescoi sp. n., P. jankowskii (Xu et al. 2009) comb. n., and P. kahli (Xu et al. 2009) comb. n., are described or redescribed based on live observation, protargol stained material, and SSU rRNA gene sequences. The new species P. paraellipticum sp. n. is characterized by its obovoidal cell shape, adoral zone composed of 17–21 collar, 9–11 buccal, and two thigmotactic membranelles, and extrusomes attached in one row along the girdle kinety. The new species P. dragescoi sp. n. is distinguished from its congeners by its obovoidal cell shape and a lack of thigmotactic membranelles. Based on ciliary patterns recognizable in the original slides, Omegastrombidium jankowskii Xu et al. 2009 and O. kahli Xu et al. 2009 should be transferred to the genus Parallelostrombidium Agatha 2004. Phylogenetic analyses based on SSU rRNA gene sequence data demonstrate that all four new sequences cluster with previously described congeners. The genus Parallelostrombidium is separated into two clusters, suggesting its non‐monophyly and probably corresponding to the two subgenera proposed by Agatha and Strüder‐Kypke (2014), as well as their morphological difference (cell dorsoventrally flattened vs. unflattened).     

Morphology,ontogeny and molecular phylogeny of a new urostylid ciliate,Bakuella (Pseudobakuella) guangdongica n. sp. (Ciliophora,Hypotrichia) from southern China

The morphology, morphogenesis, and molecular phylogeny of Bakuella (Pseudobakuella) guangdongica n. sp., isolated from southern China, were investigated. The new species is characterized by a body length of 150–225 μm in vivo; 35–42 adoral membranelles; 3–5 buccal, two frontoterminal, 7–12 transverse and two pretransverse ventral cirri; midventral complex comprised of 10–20 pairs and two rows extending to transverse cirri; posterior part of marginal rows slightly overlapping; colorless cortical granules about 1 μm across, arranged in small groups; soil habitat. Its main ontogenetic features are: (1) in the proter, the parental adoral zone of membranelles is completely renewed by new structures; (2) in the opisthe, the oral primordium originates apokinetally, some old midventral cirri join the formation of frontoventral-transverse cirral anlagen; (3) the anlagen for marginal rows and dorsal kineties develop intrakinetally; and (4) the numerous macronuclear nodules fuse into a single mass before dividing. Phylogenetic analyses based on the SSU rDNA sequence suggest the non-monophyly of the genus Bakuella.     

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,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.     

Morphology and molecular phylogeny of a new brackish pleurostomatid ciliate,Loxophyllum paludosum sp. n. (Ciliophora,Litostomatea, Haptoria), from a mangrove wetland in China

A new ciliate species of the genus Loxophyllum Dujardin, 1841, Loxophyllum paludosum sp. n., is described from a mangrove wetland near Daya Bay in Guangdong Province, southern China, based on morphological and molecular analyses. The new species can be distinguished from its congeners by a combination of the following characters: (1) 12–14 right kineties and 4–6 left kineties; (2) two macronuclear nodules and one micronucleus; (3) a single contractile vacuole located terminally; (4) extrusomes bar-shaped, evenly spaced along entire ventral margin, and clustered to form 5–7 warts along dorsal margin; and (5) presence of three ridges on the left side of cell. The new species is divergent from its congeners from 0.4% to 6.7% (5–104 nucleotide sites) based on the small subunit (SSU) rRNA gene sequence data. The validity of the new species is also supported by molecular phylogenetic trees inferred from SSU rRNA gene sequences.     

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.     

Morphology of two marine euplotids (Ciliophora: Euplotida), Aspidisca fusca and A. hexeris,with notes on their small subunit rRNA gene sequences

We investigate the living morphology, infraciliature, and small subunit rRNA gene sequences of two poorly known marine euplotids, Aspidisca fusca Kahl, 1928 and A. hexeris Quennerstedt, 1869, isolated from intertidal sewage outfall, northern China and sandy sediments, southern China, respectively. Improved diagnoses and morphometric data are provided for both species. Aspidisca fusca is characterized by having a peristomial spur, seven frontoventral cirri in a “polystyla-arrangement”, and two oval macronuclear nodules. Aspidisca hexeris can be recognized by having an elliptical body shape, a peristomial spur, seven membranelles in the anterior portion of adoral zone and 11–13 in the posterior part, as well as seven frontoventral cirri arranged in two oblique rows. In addition, the presence and degree of projections along the left margin of A. hexeris are highly variable among populations. Phylogenetic analyses based on small subunit rRNA gene sequence data support the validity of both A. hexeris and A. fusca as distinct species, and indicate that A. fusca is most closely related to A. aculeata and A. steini, whereas A. hexeris shows a close relationship with A. magna.     

Morphology,Morphogenesis and Molecular Phylogeny of a New Obligate Halophile Ciliate,Schmidtiella ultrahalophila gen. nov., spec. nov. (Ciliophora,Hypotrichia) Isolated from a Volcanic Crater on Sal (Cape Verde Islands)

A new hypotrichous ciliate, Schmidtiella ultrahalophila gen. nov., spec. nov., was isolated from a solar saltern on the island of Sal, Cape Verde. The possession of only one short dorsal kinety clearly distinguishes S. ultrahalophila from other known hypotrichous genera and species. Further diagnostic characters include: a flexible and slender body, an average size of 85 × 15 μm in vivo; a bipartite adoral zone with two hypertrophied frontal adoral membranelles and nine to twelve ventral adoral membranelles; three frontal, one parabuccal, two frontoventral, two or three postoral ventral, and two or three frontoterminal cirri; and marginal cirral rows variable in number, usually one on each side. Ontogenetic data indicate the following: the frontal‐ventral cirri originate from six or five anlagen; the proter inherits the parental adoral zone; the frontal and ventral cirri originate from five or six anlagen; and the marginal cirral rows and the dorsal kinety tend to originate intrakinetally. Additional marginal rows are rarely derived from de novo anlagen. Based on its morphology, morphogenesis and its SSU rRNA phylogenetic placement, the new species should be assigned to the order Sporadotrichida Fauré‐Fremiet, 1961. Due to low taxon sampling, however, its exact position in this order remains enigmatic.     

Ontogenesis and molecular phylogeny of a new marine urostylid ciliate, Anteholosticha petzi n. sp. (Ciliophora, Urostylida)

A new urostylid ciliate, Anteholosticha petzi n. sp., isolated from the northern China seas, was studied using live observation and protargol impregnation. It differs from all the congeners mainly in the highly flexible and variable body shape, its red-brown to brick-red colour, and the possession of three types of cortical granules. Analysis of morphometric and molecular data confirm the validity of A. petzi as a distinct species. Anteholosticha petzi n. sp. is characterized as follows: by having 3 frontal, 1 buccal, 2 frontoterminal, 2 pretransverse, and 8-11 transverse cirri; a midventral row that comprises 10-16 cirral pairs and extends ca. 60% of cell length; and 3 bipolar dorsal kineties. Ontogenesis is similar to that in Anteholosticha warreni except for the formation of the oral primordium and is characterized by: (1) the parental adoral membranelles are completely renewed and the oral primordium is formed de novo; and (2) the anlagen for the frontal-ventral-transverse cirri are formed as primary primordia. The small subunit rRNA gene sequence of A. petzi n. sp. was also compared with that of its congeners with the sequence differences ranging from 0.39% to 8.62%. Phylogenetic trees based on the SSU rRNA gene sequence were constructed, indicating the nonmonophyly of the genus Anteholosticha, as supported by the approximately unbiased test.     

Morphology and Molecular Phylogeny of Two New Brackish Water Ciliates of Bakuella (Ciliophora: Urostylida: Bakuellidae) from South Korea

Two new species of Bakuella, B. (B.) incheonensis n. sp. and B. (Pseudobakuella) litoralis n. sp., were discovered in brackish water near Aamdo Shore Park, Incheon, South Korea. We conducted a morphological study based on live observations and protargol‐impregnated specimens, and a molecular analysis using nuclear SSU rRNA gene sequences. Bakuella (B.) incheonensis is diagnosed by: body size 70–105 × 20–40 μm in vivo, 21–25 adoral membranelles, three or four frontoterminal cirri, midventral complex composed of 7–10 midventral pairs with one or two rows and terminating at about 62% of body length, 20–28 left and 25–32 right marginal cirri, 58–87 macronuclear nodules, and yellowish cortical granules. Bakuella (Pseudobakuella) litoralis is diagnosed by: body size 90–125 × 30–40 μm in vivo, 25–33 adoral membranelles, 3–5 buccal cirri, midventral complex composed 10–15 midventral pairs with one or two rows and terminated at 70% of body length, one or two pretransverse cirri, 3–6 transverse cirri, 26–39 left and 29–47 right marginal cirri, 49–84 macronuclear nodules, and two types of cortical granules. Molecular phylogeny using SSU rRNA gene sequences shows a nonmonophyletic relationship among Bakuella species and emphasizes the need for further morphogenetic studies of this genus and other related hypotrichs.     

The morphology, ontogeny, and small subunit rRNA gene sequence analysis of Diophrys parappendiculata n. sp. (Protozoa, Ciliophora, Euplotida), a new marine ciliate from coastal waters of southern China

The morphology, morphogenesis, and phylogeny of Diophrys parappendiculata n. sp., a large marine ciliate isolated from the coastal waters of Daya Bay, southern China, were investigated. This new species is characterized by a combination of its large size, appendiculata-pattern of ciliature, and bipartite adoral zone of membranelles. The main stages of morphogenesis during binary fission were also recorded and described. Comparisons of morphological characteristics with similar congeners support the validity of the new species. The small subunit rRNA gene sequence of D. parappendiculata is 96.3-99.94% similar to those of four other congeners; it differs in four nucleotides from that of Diophrys appendiculata (i.e. structural similarity was 99.94%). Phylogenetic analysis indicates that D. parappendiculata n. sp. falls into the Diophrys clade and is most closely related to the well-known D. appendiculata.     

Morphology,ontogeny and molecular phylogeny of a new brackish water ciliate Bakuella subtropica sp. n. (Ciliophora,Hypotricha) from southern China

This paper investigates the morphology, ontogenesis and small subunit (SSU) rRNA gene-based phylogeny of a new urostylid ciliate, Bakuella subtropica sp. n., discovered from the estuary of the Pearl River in Guangzhou, southern China. The new species is diagnosed by its elongate body, one buccal and one parabuccal cirrus, midventral complex comprised of 9–23 midventral pairs and one or two midventral rows extending to four fifths of body length, yellow-brown to yellow-greenish cortical granules and an estuary habitat. Its main ontogenetic features are: (1) in the proter, the parental adoral zone of membranelles is completely renewed by new structures and old midventral pairs join the formation of frontal-midventral-transverse cirral anlagen (FVT-anlagen); (2) in the opisthe, the oral primordium originates apokinetally, FVT-anlagen are formed besides and some old midventral cirri join the formation; (3) the anlagen for marginal rows and dorsal kineties develop intrakinetally; and (4) the numerous macronuclear nodules fuse into a single mass before dividing. Based on the SSU rDNA sequences, phylogenetic analyses show a close relationship between Bakuella subtropica sp. n., Apobakuella and Neobakuella, forming a clade separated from the other genera in the family Bakuellidae. Available morphological and ontogenetic data challenge the monophyly of Bakuellidae.     

Two New Endozoic Ciliates,Clevelandella lynni sp. n. and Nyctotherus galerus sp. n., Isolated from the Hindgut of the Wood‐feeding Cockroach Panesthia angustipennis (Illiger, 1801)

Two new ciliate species, Clevelandella lynni sp. n. and Nyctotherus galerus sp. n., were discovered in the hindgut of wood‐feeding panesthiine cockroaches. Their morphology was studied using standard methods, and their phylogenetic positions within the order Clevelandellida were determined using the 18S rRNA gene sequences. Clevelandella lynni is characterized by a prominent peristomial projection, a notched left body margin, a tear‐shaped to broadly ovoidal macronucleus, a karyophore attached to the right body margin, and by an adoral zone composed of on average 48 membranelles and extending about 51% of body length. The diagnostic features of N. galerus include a short posterior body projection, a spherical to broadly ellipsoidal macronucleus, a karyophore attached to the right and left body margins, refractile bodies densely packed anterior to the macronucleus, and an adoral zone composed of on average 57 membranelles and extending about 70% body length. The order Clevelandellida was consistently depicted as monophyletic in 18S rRNA gene phylogenies. Nyctotherus galerus was placed in the paraphyletic family Nyctotheridae, as sister taxon to all other Nyctotherus and Clevelandella species isolated from cockroaches. Clevelandella lynni fell in the monophyletic family Clevelandellidae, as sister taxon to C. panesthiae KC139718 but with very poor statistical support.     

A New Subspecies of Oxytricha granulifera (Hypotrichia: Oxytrichidae) from Mexico,with Notes on its Morphogenesis and Phylogenetic Position

The genus Oxytricha Bory de Saint‐Vincent in Lamouroux, Bory de Saint‐Vincent and Deslongchamps, 1824 comprises about 38 species distributed worldwide and has been considered to be a nonmonophyletic group. Based on living observations, protargol preparations, and a small subunit ribosomal RNA (SSU rRNA) gene sequence, we describe a new subspecies Oxytricha granulifera chiapasensis n. subsp. This new taxon is morphologically characterized by undulating membranes basically in a Stylonychia‐pattern, six dorsal kineties, size in vivo ca. 60–120 × 20–40 μm, 21–30 right and 21–31 left marginal cirri, 22–29 adoral membranelles, and spherical cortical granules arranged in longitudinal rows on the dorsal side. In terms of the SSU rRNA gene sequence, the new subspecies differs from populations of O. granulifera from GENBANK by 7–35 nucleotides. Phylogenetic analyses showed that Oxytricha granulifera gene sequences were nested into three groups, with the new subspecies included in one of them. Oxytricha granulifera chiapasensis n. subsp. is different from Oxytricha granulifera granulifera Foissner and Adam, 1983 and Oxytricha granulifera quadricirrata Blatterer and Foissner, 1988 based on: (i) undulating membranes in Stylonychia‐pattern, (ii) formation of a sixth dorsal kinety during morphogenesis, (iii) the adoral membranelles number, and (iv) inhabiting freshwater habitats.     

Morphology,morphogenesis, and molecular phylogeny of a new freshwater ciliate,Gonostomum jangbogoensis n. sp. (Ciliophora,Hypotricha), from Victoria Land,Antarctica

In a study on ciliate diversity, we discovered the new hypotrich species, Gonostomum jangbogoensis n. sp., in freshwater from Terra Nova Bay, Victoria Land, southeast Antarctica. We describe its morphology and morphogenesis using standard methods, and the SSU rRNA gene phylogeny is provided as well. Morphology of Gonostomum jangbogoensis n. sp. is characterized as follows: slender to elongated body shape; grayish under low magnification; cortical granules present; 32–41 adoral membranelles; 3 enlarged frontal cirri; 1 buccal cirrus; 2 frontoterminal cirri; 3 or 4 frontoventral cirral pairs, 2 pretransverse cirri, 6–7 transverse cirri; 13–19 left and 18–26 right marginal cirri; 17–23 paroral kinetids; 3 dorsal kineties; 3 caudal cirri; 2 macronuclear nodules with 1–3 micronuclei. The morphogenesis of the new species confirms that it has at least seven frontal-ventral-transverse cirral anlagen, which is also reported in Gonostomum sp. 1 sensu Shin from Korea. Even though these two populations occur very far from each other, the morphometric data prove that this character state, the seven cirral anlagen, is a stable feature across these populations and might be an apomorphy. The phylogenetic analyses show that the genus Gonostomum is non-monophyletic and that the new species is a sister to G. bromelicola.     

Morphology and Notes on Morphogenesis during Cell Division of Deviata polycirrata n. sp. and of Deviata bacilliformis (Gelei, 1954) Eigner, 1995 (Ciliophora: Kahliellidae) from Argentina

ABSTRACT. Described herein are the morphology and certain morphogenetic stages of a new freshwater ciliate species, Deviata polycirrata n. sp., and of Deviata bacilliformis recorded in the soil of a dried temporary pond from Argentina. Ciliates were studied alive and after silver impregnation with Protargol. Deviata polycirrata n. sp. measures 130–180 × 45–70 μm in vivo. The species possesses 8–9 long cirral rows on the right and 9–13 on the left of the oral zone, and 3 dorsal rows of dikinetids. The adoral zone is composed of 39–48 membranelles. There are four macronuclear nodules and usually two micronuclei. A single contractile vacuole is located equatorially on the left body margin. This new species mainly differs from its congeners in having a higher number of cirral rows, the three long dorsal rows of dikinetids (vs. usually one to two dorsal rows of dikinetids), and a higher number of adoral membranelles. The other species reported here, D. bacilliformis, is recorded for the first time in Argentina. Unlike previous observations on this species, on the dorsal surface there are cirral rows that are preceded by cilia (combined cirral rows), and stomatogenesis begins with the proliferation of non‐ciliferous basal bodies some distance posterior to the buccal vertex.     

Morphology and morphogenesis of a new marine hypotrichous ciliate (Protozoa,Ciliophora, Pseudoamphisiellidae), including a report on the small subunit rRNA gene sequence

The morphology and morphogenesis of a new marine hypotrichous ciliate Pseudoamphisiella elongata sp. nov. isolated from mussel‐farming waters near Qingdao, China, are described based on living and protargol‐impregnated specimens. Morphologically, the new species can be distinguished from its known congeners by its elongate body shape, narrow oral field, having fewer dorsal kineties and caudal cirri, more marginal cirri, and differentiated pretransverse cirri. The identification as a new species is firmly supported by the sequences of the small subunit ribosomal rRNA (SSU rRNA) gene, compared with other known Pseudoamphisiella species, and the phylogenetic analysis. The morphogenetic characteristics can be summarized as follows: (1) the parental adoral zone of membranelles and undulating membranes are entirely rebuilt by the oral primordium, which develops de novo in the outermost region of the cortex; (2) the oral primordium in the opisthe and the frontoventral–transverse (FVT) anlagen in both dividers are formed independently on the cell surface; (3) an ‘extra’ marginal anlage originates to the right of the right marginal anlage, and develops into two or three ‘extra’ marginal cirri; (4) the FVT anlagen develop in the primary mode, and the last FVT streak contributes two migratory cirri (frontoterminal cirri), which are probably resorbed; (5) the right marginal anlagen in both dividers occur close together, independent of the old structure. © 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 158 , 231–243.     

Morphology,morphogenesis and molecular phylogeny of Lamtostyla granulifera sinensis subsp. nov. (Ciliophora,Hypotrichia) from a wetland in China

The morphology, morphogenesis and molecular phylogeny of a hypotrichous ciliate, Lamtostyla granulifera sinensis subsp. nov., isolated from northern China, were investigated. This population appeared highly similar in morphology to L. granulifera Foissner, 1997. However, on detailed investigation some non-overlapping features were identified, i.e., the body shape and the arrangement of the cortical granules. These differences suggested the separation at subspecies level. Furthermore, the morphogenesis of the new subspecies is described, which is characterized by: (1) the posterior part of the parental adoral zone of membranelles is renewed; (2) the amphisiellid median cirral row is formed from two anlagen; and (3) the frontoventral-transverse cirral anlagen II to VI generate one transverse cirrus each. Phylogenetic analyses based on SSU rDNA sequence data show that Lamtostyla species are scattered in different clades. The monophyly of the genus Lamtostyla is also rejected by the AU test in this study.