Hongbo Pan

Descriptions of two new marine scuticociliates, Pleuronema sinica n. sp. and P. wilberti n. sp. (Ciliophora: Scuticociliatida), from the Yellow Sea, China

Two new marine scuticociliates, Pleuronema sinica n. sp. and P. wilberti n. sp., collected from the sand beach of Qingdao, China, were investigated in vivo and following protargol impregnation. Ciliates of the genus Pleuronema are normally recognizable by their large sail-like paroral membrane although one species, P. grolierei, has shorter cilia in the paroral membrane. Neither of the new forms has the conspicuous paroral membrane in vivo so in this respect they are not typical members of this genus. Pleuronema sinica is characterized by its large, conspicuously flattened body, the possession of only one preoral kinety, the irregular-shaped macronucleus and the rather unusual structure of the oral apparatus. By contrast P. wilberti has a medium-size broad-oval body, six to eight preoral kineties and a highly differentiated membranelle 3 that is five- or six-rowed. An identification key is supplied for the 15 species of Pleuronema for which the infraciliature is known.

Three New Loxophyllum Species (Ciliophora: Pleurostomatida) from China with a Brief Review of the Marine and Brackish Loxophyllum Species

Recent studies indicate that there is a high diversity of pleurostomatid ciliates in the coastal waters of China. Here, three new congeners of Loxophyllum, L. caudatum sp. n., L. rugosum sp. n., and L. chinense sp. n., are described following observations of live cells and protargol‐impregnated specimens. All three species usually have two macronuclear nodules and prominent warts along the dorsal margin formed by clustered extrusomes. In addition, L. caudatum sp. n. is characterized by its long conspicuous tail, dot‐like cortical granules, 4 or 5 left and 9 or 10 right kineties, and a single subterminal contractile vacuole. Loxophyllum rugosum sp. n. is distinguished by possessing three prominent ridges on the left side, 7–11 right and 5–7 left kineties. Loxophyllum chinense sp. n. is characterized by having several contractile vacuoles distributed along the ventral margin, 13–18 right and 6–8 left kineties. The small subunit ribosomal DNA (SSU rDNA) sequence similarities among six congeners range from 96.46% to 99.94%. Phylogenetic trees based on the SSU rDNA sequences indicate that all Loxophyllum spp. form a well‐supported monophyletic group. A brief review of the marine and brackish Loxophyllum species is supplied and one new combination, Litonotus multiplicatus (Kahl 1931) comb. n. (basionym Loxophyllum multiplicatum Kahl 1931), and one new name, Litonotus dragescoi nom. n. (basionym L. fasciolatus Dragesco 1966), are suggested.

Morphology and Molecular Phylogeny of Three Cyrtophorid Ciliates (Protozoa, Ciliophora) from China, Including Two New Species, Chilodonella parauncinata sp. n. and Chlamydonella irregularis sp. n.

This study investigated the morphology and molecular characteristics of three interesting free‐living cyrtophorid ciliates, including two new species, isolated from China: Chilodonella parauncinata sp. n. can be identified by its elongated body shape, with a sharp protrusion in the left anterior part, cell size ca. 60 × 25 μm in vivo, five right and 6–7 left kineties with kinetosomes densely arranged, and a curved cyrtos. Chlamydonella irregularis sp. n. differs from its congeners by the oval body shape, cell size 50–60 × 25–40 μm in vivo, irregular shape of macronucleus, 30–40 club‐shaped ventral protuberances, and 17 somatic kineties. Two isolates of Chlamydonella derouxi Song, 2003, collected from an intertidal area in Shandong and a mangrove wetland in Guangdong respectively, correspond well with two previous descriptions, but differ in comprising more basal bodies in left and right equatorial fragments and in having more finger‐like protuberances on the ventral side. Phylogenetic analyses based on the small subunit rRNA gene sequences showed that C. parauncinata sp. n. clustered with Chilodonella uncinata, but was a well‐outlined species of the genus, and C. irregularis sp. n. and C. derouxi grouped in the family Lynchellidae with their congeners to form the monophyletic genus Chlamydonella.

Morphological and Molecular Description of Three New Species of the Cyrtophorid Genus Chlamydodon (Ciliophora, Cyrtophoria)

This study investigates the morphology and molecular characteristics of three new cyrtophorid ciliates isolated from China seas: Chlamydodon salinus n. sp., Chlamydodon caudatus n. sp., and Chlamydodon paramnemosyne n. sp. Of these, C. salinus n. sp. differs from its congeners through a combination of body size, a cross‐striated band that is not continuous, the presence of 30–34 somatic kineties, 11–15 nematodesmal rods, and 13 contractile vacuoles. Chlamydodon caudatus n. sp., meanwhile, is characterized by having a conspicuous tail, a continuous cross‐striated band, 34–40 somatic kineties, about 15 contractile vacuoles, and 20–24 nematodesmal rods. Compared with other Chlamydodon species, the third new one, C. paramnemosyne n. sp., could be identified by its continuous cross‐striated band, 16–18 somatic kineties, 5 contractile vacuoles, and 9–12 nematodesmal rods. Based on the sequence of the small subunit (SSU) rRNA gene, the phylogeny of these three new species was analyzed, indicating that they all clustered with other congeners to form a monophyletic assemblage. Based on previous studies and the present work, a brief revision of the genus Chlamydodon is supplied, and a key to aid the identification of Chlamydodon species is given.

Diversity of free-living marine ciliates (Alveolata, Ciliophora): Faunal studies in coastal waters of China during the years 2011–2016

In the period 2011-2016, a series of investigations were carried out on the marine and brackish free-living ciliate fauna of the temperate-tropical coastal waters of China. About 210 morphotypes including over 100 new species within six groups (cyrtophorians, hypotrichs s.l., karyorelicteans, oligotrichs, pleurostomatids, and scuticociliates) were isolated and described in detail from observations of live cells and silver-stained specimens. Based on their morphology, morphogenesis and molecular phylogeny, three new families (Wilbertomorphidae, Kentrophyllidae, Protolitonotidae) and 22 new genera (Apotrachelocerca, Wilbertomorpha, Protolitonotus, Paracyrtophoron, Heterohartmannula, Aporthotrochilia, Falcicyclidium, Paramesanophrys, Pseudodiophrys, Monocoronella, Neourostylopsis, Apobakuella, Parabistichella, Heterokeronopsis, Heterotachysoma, Antiokeronopsis, Apoholosticha, Pseudogastrostyla, Antestrombidium, Sinistrostrombidium, Williophrya, and Varistrombidium) were established. In the present review, we summarize these studies which show there is a large, undiscovered diversity of ciliates, especially in undersampled habitats, such as subtropical/tropical coastal waters, mangrove wetlands, estuaries and aquaculture ponds. We also highlight the importance of integrative approaches, combining morphology, morphogenesis and molecular phylogeny, in order to understand ciliate systematics and ecosystem function.

Morphological redescriptions of four marine ciliates (Ciliophora: Cyrtophorida: Dysteriidae) from Qingdao, China

The morphology and infraciliature of four marine cyrtophorid ciliates isolated from Qingdao, China, were investigated. Based on the present work and on previous data, improved diagnoses for three rarely known species are provided: (1) Mirodysteria decora; small-sized marine Mirodysteria about 35-60 × 25-35 μm in vivo, oval in outline; body surface with two or three conspicuous dorsal spines and one caudal spine; three right kineties, the rightmost one extending dorso-apically; left frontal kineties reduced, each consisting of three basal bodies only; podite subcaudally positioned; two ventrally located contractile vacuoles. (2) Dysteria legumen; body oval with two longitudinal grooves on different plates; six right kineties, the rightmost two of which extend dorso-apically; two left frontal kineties and two ventrally located contractile vacuoles. (3) Dysteria proraefrons; body about 60×35 μm in vivo; six right kineties, the two rightmost of which extend dorso-apically and the leftmost one is considerably shortened; three left frontal kineties; two ventrally located contractile vacuoles. A population of D. derouxi with eight or nine right kineties is also briefly described. The current investigation further demonstrates high diversity and cosmopolitan distribution of this highly specialized group of benthic ciliates.

New considerations on the phylogeny of cyrtophorian ciliates (Protozoa, Ciliophora): expanded sampling to understand their evolutionary relationships

To rationalize the confusing relationships among the cyrtophorian ciliates, we expanded the taxon sampling by sequencing the small subunit ribosomal RNA (SSU rRNA) gene of representatives of 12 genera (20 species, 23 new sequences). The SSU rRNA sequences of Spirodysteria, Agnathodysteria, Brooklynella and Odontochlamys are reported for the first time. Phylogenetic trees were constructed, and secondary structures of variable region 4 (V4) of all genera for which SSU rRNA gene sequence data are available were predicted. The results indicate that (i) Brooklynella is likely an intermediate taxon between Dysteriidae and Hartmannulidae; (ii) the genus Dysteria is paraphyletic with Spirodysteria and Mirodysteria nested within it; (iii) the genus Agnathodysteria is well separated from Dysteria based on both molecular and morphological data; and (iv) Trithigmostoma is a basal genus of Chilodonellidae, based on both the morphological and molecular data.

Morphology and phylogeny of two species of Loxodes (Ciliophora, Karyorelictea), with description of a new subspecies, Loxodes striatus orientalis subsp. n.

The morphology and phylogeny of Loxodes vorax and L. striatus orientalis subsp. n. were investigated based on infraciliature and small subunit (SSU) rRNA gene sequence data. Loxodes striatus orientalis subsp. n. was separated from L. striatus striatus stat. n. by having fewer dikinetids in the intrabuccal kinety (35–55 vs. 50–70) and a variable number of macronuclei (2–4 vs. 2). In addition, the SSU rRNA gene sequence of the new subspecies differs in 13 and 11 nucleotides from that of two populations of the nominotypic subspecies. We also summarized the morphological differences between Loxodes and Remanella based on the data available. Phylogenetic analyses revealed that the genus Loxodes was monophyletic and nested within Remanella species. This study might, therefore, support the hypothesis that the freshwater genus Loxodes evolved from the marine genus Remanella.

Morphological and phylogenetic studies on three members of the genus Pseudochilodonopsis (Ciliophora, Cyrtophoria) isolated from brackish waters in China, including a novel species, Pseudochilodonopsis quadrivacuolata sp. nov.

Three cyrtophorian ciliates isolated from brackish biotopes in China, Pseudochilodonopsis quadrivacuolata sp. nov., Pseudochilodonopsis fluviatilisFoissner, 1988 and Pseudochilodonopsis mutabilisFoissner, 1981, were investigated using living observation and protargol-staining methods. P. quadrivacuolata sp. nov. can be characterized as follows: cell size 50-70 × 30-40 μm in vivo; body oval with posterior end rounded; four tetragonally positioned contractile vacuoles; 12-15 nematodesmal rods; five right and six left somatic kineties; terminal fragment positioned apically on dorsal side, consisting of 11-14 basal bodies; four or five fragments in preoral kinety. P. fluviatilis and P. mutabilis were generally consistent with previous descriptions. In addition, a brief revision and a key to Pseudochilodonopsis are presented. The small-subunit (SSU) rRNA gene was also sequenced to support the identification of these species. Phylogenetic analyses based on molecular data indicate that the genera Pseudochilodonopsis and Chilodonella are closely related and both are well outlined; that is, all known congeners for which SSU rRNA gene sequence data are available group together, forming the core part of the family Chilodonellidae.

Morphology of Three Species of Amphileptus (Protozoa, Ciliophora, Pleurostomatida) from the South China Sea, with Note on Phylogeny of A. dragescoi sp. n.

Two new and one problematic species of pleurostomatids, Amphileptus dragescoi sp. n., A. wilberti sp. n., and A. marinus from coastal areas of the South China Sea, are described based on observations of live and protargol‐impregnated specimens. Amphileptus dragescoi is different from its congeners by the presence of an apical group of extrusomes and the possession of 12–15 right and five left somatic kineties, two macronuclear nodules, and a single terminally positioned contractile vacuole. Amphileptus wilberti is diagnosed by oval or pyriform body, 15–19 right and seven or eight left somatic kineties, extrusomes arranged only in anterior portion of oral slit, usually three ventrally located contractile vacuoles, and two macronuclear nodules. Amphileptus marinus (Kahl, 1931) Song et al., 2004 is redescribed and its diagnosis is improved. One isolate which was misidentified as A. marinus by Song et al. (2004) is believed to represent an unknown form, named here as Amphileptus songi sp. n. Phylogenetic analyses of the SSU rDNA sequences indicate that the genus Amphileptus is paraphyletic, but its monophyly is not rejected by statistical tree topology tests.