Paroral Membrane Research Articles

Identification of Boveria labialis parasitism in the respiratory tree of the Cultured Sea cucumber, Apostichopus japonicus

Aquaculture of the sea cucumber Apostichopus japonicus has emerged as a pivotal industry in northern China in the early 21st century. It is estimated that the cultivation area for this species spans 250,356 ha, yielding an annual production of 248,508 tons. The swift growth and intensification of sea cucumber farming practices have triggered a spectrum of diseases, resulting in significant economic detriment. This study offers an exhaustive account of a particular ciliate species, identified through smear and histopathological observations, hematoxylin-eosin staining, scanning electron microscopy, and molecular biology techniques, detailing its morphology, size, cellular architecture, and reproductive mode. The ciliate exhibits a torch-like shape, measuring 40–82 μm in length and 12–35 μm in width. It is characterized by the presence of both a macronucleus and a micronucleus. The anterior end is slender and blunt, while the posterior end is the widest part of the organism. A peristomal disc (PD) is situated at the body's terminus, with the mouth positioned centrally on the PD and a lip-like projection (6–10 μm in length) extending from one side. The organism's surface, particularly the area surrounding the oral region, is adorned with an abundance of long cilia. A single contractile vacuole is observable at the aboral end of the ciliate. It possesses 17–21 somatic kineties, complemented by 3 membranelles (M1, M2, M3) and a paroral membrane (PM) within the buccal cavity. One contractile vacuole was seen at the aboral end. There were 17–21 somatic kineties, while three membranes (M1, M2, M3) and a paroral membrane (PM) located within the buccal cavity. The first somatic kinety is notably more extended than the others, uniquely reaching a length of 10 μm around the PM. The PM, comprised of 2 somatic kineties in a “zig-zag” formation, follows a spiral path, making approximately 360-degree turns in conjunction with M1, M2, and M3, and culminates at the mouth depression. The ciliate reproduces through transverse binary fission or sexual conjugation. Based on these distinguishing features and molecular biological identification results, the ciliate is provisionally classified within the genus Boveria, as Boveria labialis. Histopathological examination has shown that these ciliates are predominantly found within the respiratory trees of their hosts, where they inflict damage to the epithelial tissues. This damage manifests as cell necrosis, structural disintegration, and the shedding of cells from the respiratory tree's epithelial lining. In severe cases, the affected respiratory tree and other viscera may be expelled. Epidemiological studies have indicated that infected individuals do not present with any overt external lesions. The findings of this research are instrumental for the management of diseases and the promotion of health within the sea cucumber aquaculture industry.

Exploring the biogeography, morphology, and phylogeny of the condylostomatid ciliates (Alveolata, Ciliophora, Heterotrichea), with establishment of four new Condylostoma species and a revision including redescriptions of five species found in China

Species of the ciliate class Heterotrichea Stein, 1859 are a cosmopolitan group of unicellular eukaryotic microorganisms, many of which have been widely used as models in various fields of research such as regenerative biology, functional ecology, environmental toxicology, and symbiotic behavior. However, species identification in the heterotrich family Condylostomatidae, especially the most species-rich and type genus Condylostoma Bory de Saint-Vincent, 1824, remains challenging due to incomplete original descriptions, few reliable distinguishing characters, and overlapping features between different species. This study presents an updated revision of Condylostoma and its related genus Condylostomides da Silva Neto, 1994 based on descriptions of five species, including nine populations collected from China, using both morphological and molecular methods. The main findings are as follows: (1) 43 nominal species and about 130 populations are reviewed, resulting in the recognition of 30 valid species of Condylostoma and eight valid species of Condylostomides; (2) keys, synonyms, biogeographic distributions and amended/improved diagnoses of all valid species are provided; (3) based on the available data, four new Condylostoma species (C. marinum sp. nov., C. petzi sp. nov., C. villeneuvei sp. nov., and C. microstomum sp. nov.), one new combination (Condylostomides minimus (Dragesco, 1954) comb. nov. & nom. corr.), and two corrected names (Condylostoma ancestrale Villeneuve-Brachon, 1940 nom. corr. and Condylostomides nigrus (Dragesco, 1960) nom. corr.) are suggested; (4) cryptic species are detected and proposed for the first time to form the Condylostoma curvum species complex; (5) three highly confusing Condylostoma species, C. kris, C. spatiosum, and C. minutum, are redefined for the first time based on modern taxonomic methods; (6) a ‘flagship’ species, Condylostomides coeruleus, is recorded for the first time from the continent of Asia, substantially expanding its biogeography; (7) ciliature adjacent to the distal end of the paroral membrane within the family Condylostomatidae is uniformly defined as frontal membranelles and is classified into three patterns according to the arrangement of kinetosomes, which serve as important key features.

Ontogeny and phylogeny of the anaerobic genus Urostomides with supplementing morphological characterization of the type and one little-known species (Alveolata: Ciliophora: Armophorea)

Abstract Urostomides is an obligate anaerobic ciliate genus in the family Apometopidae (Metopida). Although most congeners have been described using modern criteria, detailed ontogenetic data for both the family and genus have not been reported yet. In this study, we investigate the morphogenesis of two Urostomides species, viz., U. minimus (Kahl, 1927) comb. nov. and U. striatus (type species) in detail. Our findings reveal that the process closely resembles that of several described Metopidae species, except for a notable difference in the origin of the new adoral membranelles either from all perizonal rows or from other somatic kineties. Additionally, the data of U. minimus on the ciliature and SSU rRNA gene sequence are provided and those of U. striatus are supplemented based on Chinese isolates. We show the diplostichomonad morphology of the paroral membrane, a feature not previously observed in Urostomides species. Furthermore, our phylogenetic analyses confirm the monophyly of Urostomides.

Morphogenesis of an anaerobic ciliate Heterometopus palaeformis (Kahl, 1927) Foissner, 2016 (Ciliophora, Armophorea) with notes on its morphological and molecular characterization

The morphology, morphogenesis, and molecular phylogeny of Heterometopus palaeformis (Kahl, 1927) Foissner, 2016 were studied using microscopical observations on live and protargol-stained specimens as well SSU rRNA gene sequencing. The morphogenetic data for the genus are presented for the first time. Compared to other metopids, the morphogenesis of H. palaeformis is distinct since its (1) perizonal stripe rows 4 and 5 are involved in the formation of the opisthe’s adoral polykinetids; (2) perizonal stripe rows 3–5 and two adjacent preoral dome kineties contribute to most of the opisthe’s paroral membrane while perizonal stripe rows 1 and 2 contribute very little; (3) four kinety rows are formed to the left of the opisthe’s adoral zone of polykinetids. The Chinese population resembles the original and neotype populations well in terms of general morphology — characterized by a life size of 55–120 × 10–20 μm, an elongate ellipsoidal body with a hardly spiralized flat preoral dome, about 18 somatic kineties and 20 adoral polykinetids. The SSU rDNA sequence of the present population exhibits a disparity of 1.33%–2.22% divergence from sequences of other populations. Nevertheless, phylogenetic analysis reveals that populations of H. palaeformis form a separate, stable cluster within the paraphyletic Metopidae clade.

Morphology, morphogenesis, and molecular characterization of Castula specialis sp. nov. (Ciliophora, Armophorea, Metopida).

The morphology, morphogenesis, and molecular phylogeny of a new metopid ciliate, Castula specialis sp. nov., comprising three strains from geographically distant (China, Mexico, Czech Republic) anoxic freshwater habitats, were studied based on microscopic observation of live and protargol-stained specimens as well as SSU rRNA gene sequence data. The new species is characterized as follows: size invivo 105-220 × 25-70 μm, body oblong to elongated ellipsoidal and asymmetrical; preoral dome distinctly projecting beyond the body; 32-46 adoral membranelles; 31-52 somatic kineties; and 4-7 setae. This study brings the first morphogenetic investigation of a member of the genus Castula. The morphogenesis of the type population (China) of the new species proceeds as in Metopus spp. comprising drastic changes in body shape and a pleurotelokinetal stomatogenesis; however, the main difference is the origin of the opisthe's paroral membrane that derives from all perizonal rows and some adjacent dome kineties. Phylogenetically, the genus Castula is paraphyletic.

The search finds an end: the morphologically chimeric hysterocinetids belong to the subclass Hymenostomatia (Ciliophora: Oligohymenophorea)

Abstract The digestive tract of oligochaetes harbours a diverse ciliate community embracing plagiotomids, nyctotherids, astomes, and hysterocinetids. Although several hundred intestinal ciliate species are known, only two hysterocinetids have been reported from the intestine of Holarctic lumbricid earthworms hitherto. The present study is focused on the taxonomy, morphology, and molecular phylogeny of two lumbricid-dwelling taxa, the insufficiently known Protoptychostomum simplex and Hysterocineta bellerophon sp. nov., detected in Central Europe. Diagnoses of the genera Protoptychostomum and Hysterocineta were improved taking into account also features of the oral ciliature for the first time. In the past, the proper classification of hysterocinetids in the class Oligohymenophorea was hampered by the dramatic remodelling of the anterior body end into a thigmotactic sucker, as well as by the helicalization and transfer of the oral apparatus to the posterior body pole. The present phylogenetic analyses of nuclear and mitochondrial genes robustly reject the classification of hysterocinetids in subclass Scuticociliatia or as a distinct subclass, Hysterocinetia. However, hysterocinetids were consistently and robustly nested deep in subclass Hymenostomatia in a sister-position to the free-living Tetrahymena paravorax. The hymenostome affinity of hysterocinetids is also corroborated by the parakinetal stomatogenesis and the stichodyad paroral membrane accompanied by three regularly organized membranelles.

Re-discovery and novel contributions to morphology and multigene phylogeny of Protospirella mazurica (Raabe, 1968) Aescht, 2001 (Ciliophora: Pleuronematida), an obligate symbiont of the river nerite Theodoxus fluviatilis Linnaeus, 1758 (Mollusca: Gastropoda)

Although the river nerite Theodoxus fluviatilis (Gastropoda: Neritimorpha) has an exceptionally broad geographical and ecological distribution, it carries only four ciliate species: Hypocomella quatuor, Protospirella mazurica, Scyphidia sp., and Trichodina baltica. After more than a half-century gap, we re-discovered P. mazurica in a Danubian population of T. fluviatilis (haplotypes F31 and F34) and characterized it using an integrative morpho-molecular approach. Protospirella mazurica is distinguished by (i) a small, elongate-ellipsoidal to ovoidal body, (ii) a broadly ellipsoidal macronucleus accompanied by a single globular micronucleus, (iii) a subterminal contractile vacuole, (iv) about 24 somatic kineties, (v) thigmotactic ciliature composed of about 10 kineties shortened posteriorly to form a parenthetical system, and (vi) a long inverted J-shaped paroral membrane associated with three unequally long membranelles. According to the present phylogenetic analyses of two mitochondrial and three nuclear markers, P. mazurica robustly clusters within the order Pleuronematida (Oligohymenophorea: Scuticociliatia) along with other symbiotic members of the families Hemispeiridae and Thigmophryidae as well as free-living representatives of the paraphyletic family Cyclidiidae. In light of the present phylogenetic analyses, we consider the family Ancistridae to be a junior synonym of the family Hemispeiridae, which collates 14 genera in our classification framework.

Morphology and molecular phylogeny of three freshwater scuticociliates, with establishments of one new genus and three new species (Ciliophora, Oligohymenophorea)

The morphology and molecular phylogeny of three freshwater scuticociliates, Variuronema elegans gen. nov., spec. nov. (type species), V. similis spec. nov. and Uronema rigidum spec. nov., collected from Qixing River Wetland, northeastern China, were investigated using live observation and silver carbonate staining methods; the phylogenetic placements were inferred from the small subunit rRNA gene. Variuronema gen. nov., assigned to the family Uronematidae, is distinguished from the other genera mainly by its membranelles 1 and 2, each consisting of four longitudinal rows of kinetids, and the paroral membrane extending anteriorly to the front part of membranelle 3. The phylogenetic analyses indicate that: (1) Variuronema spp. form an independent, fully supported clade. However, its position among other uronematids is unresolved. (2) The family Uronematidae is not monophyletic based on the SSU rRNA gene, which is in accordance with previous studies.

Taxonomy and molecular phylogeny of two new Blepharisma species (Ciliophora, Heterotrichea) from northeastern China

The morphology and ciliary pattern of two new heterotrich ciliates, Blepharisma orientale n. sp. and B. sinicum n. sp., collected from Sanjiang Plain Wetland, northeastern China, were investigated using living observation and protargol staining. Blepharisma orientale n. sp. is characterized by the following combination of characteristics: limnetic; body in vivo 280–380 × 120–190 μm, slender to elongate, cortical granules dark pink, ventral margin somewhat sigmoid; buccal field about 55–60% of body length; 30–39 somatic kineties; 70–76 adoral membranelles, each comprising two long rows of basal bodies; paroral membrane consists of dikinetids with the posterior third obliquely arranged, both basal bodies in each dikinetid are ciliated. Blepharisma sinicum n. sp. is recognized by the combination of the following characters: limnetic; body 190–260 × 100–170 μm in size in vivo, irregularly sigmoid and slightly bilaterally flattened, dorsal margin curved; cortical granules pale pink in color; buccal field about 60–75% of body length; 25–29 somatic kineties; 53–82 adoral membranelles; paroral membrane consisting entirely of dikinetids. Phylogenetic analyses based on small subunit rRNA (SSU rRNA) gene sequence data recovered the two new species as sister taxa, both the sister group of B. penardi.

Diversity and Phylogenetic Position of Bothrostoma Stokes, 1887 (Ciliophora: Metopida), with Description of Four New Species

Bothrostoma is a genus of anaerobic ciliates in family Metopidae comprising four species, all described based solely on the morphology of living and fixed cells. Unlike other metopids, cells of Bothrostoma are not twisted anteriorly, have a flattened preoral dome, a very prominent sail-like paroral membrane, and an adoral zone of distinctive, very narrow, curved membranelles confined to a wide, non-spiraling peristome on the ventral side. We examined 20 populations of Bothrostoma from hypoxic freshwater sediments. We provide morphological characterization and 18S rRNA gene sequences of four new species, namely B. bimicronucleatum sp. nov., B. boreale sp. nov., B. kovalyovi sp. nov., and B. robustum sp. nov., as well as B. undulans (type species), B. nasutum, and B. ovale comb. nov. (original combination Metopus undulans var. ovalis Kahl, 1932). Except for B. nasutum, Bothrostoma species show low genetic variability among geographically distant populations. Intraspecific phenotypic variability might be driven by environmental conditions. In phylogenetic analyses, Bothrostoma is not closely related to Metopus sensu stricto and forms a moderately supported clade with Planometopus, here referred to as BoPl clade. The anterior axial torsion of the body, typical of other Metopidae, appears to have been lost in the last common ancestor of the BoPl clade.

Morphology, morphogenesis and phylogeny of Anteglaucoma orientalis n. sp. (Ciliophora, Oligohymenophorea), a freshwater hymenostomatid from northeastern China

A new hymenostomatid ciliate, Anteglaucoma orientalis n. sp., isolated from a freshwater pond in Harbin, northeastern China, was investigated using live observation and silver staining methods. Anteglaucoma orientalis is characterized as follows: size in vivo about 50–60 × 30–35 μm; oval body shape; buccal area occupies about 25% of body length; 28–36 somatic kineties; membranelle 1 having six or seven basal body rows, membranelle 2 five to seven rows, and membranelle 3 three rows; single macronucleus with one micronucleus attached. Morphogenesis of the genus Anteglaucoma is revealed for the first time. The main events during binary fission are as follows: morphogenesis begins with proliferation of kinetosomes in the middle part of postoral kinety 1, and kinetosomes of this primordial field multiply and organize to finally form the paroral membrane and membranelles 1–3 of the opisthe; the parental apparatus in the proter does not take part in the stomatogenetic process. Phylogenetic analyses based on SSU rRNA gene sequences show that Anteglaucoma orientalis n. sp. clusters with the type species, A. harbinensis Pan et al., 2017, with full support.

Morphology and phylogeny of a new soil ciliate, Colpodidium zelihayildizi n. sp. (Ciliophora, Nassophorea, Colpodidiidae), from Van, Turkey

new ciliate species Colpodidium zelihayildizae n. sp., which was discovered in a dry irrigation channel in Van (Turkey), was investigated morphologically using standard live and silver impregnation methods, and its SSU rRNA gene phylogeny was provided. This new species was characterized as follows: body size about 40-60 × 15-20 ?m in vivo, elongated ellipsoidal with flat or slightly concave ventral surface; the buccal cavity slightly below equatorial plane; macronucleus spherical or slightly elliptical, usually located in the last third of the cell; cortex distinctly furrowed in the anterior half; 19-22 somatic kineties, 4 postoral kineties; paroral membrane has 20 dikinetids; 3 nassulid organelles (NOs), with NO1 very close to the left edge of the paroral membrane, which consists of 1 dikinetid pair. Phylogenetic analyses conducted on the SSU rDNA sequences determined that Colpodidium zelihayildizae was clustered with Colpodidium caudatum and formed a clade with Apocolpodidium etoschense and Colpodididae sp.

Re-discovery and novel contributions to morphology and multigene phylogeny ofMyxophyllum steenstrupi(Ciliophora: Pleuronematida), an obligate symbiont of terrestrial pulmonates

AbstractMyxophyllum steenstrupi is a symbiotic ciliate living in the body slime and mantle cavity of terrestrial pulmonates (Gastropoda: Pulmonata). In the present study, M. steenstrupi was re-discovered after almost 30 years and characterized using an integrative morpho-molecular approach for the first time. Myxophyllum is distinguished by a broadly ovate, about 140 × 115 μm-sized body, a nuclear apparatus typically composed of seven macronuclear nodules and a single micronucleus, a central contractile vacuole, a shallow oral cavity situated in the posterior body region and dense somatic ciliature with extensive thigmotactic field. According to the present phylogenetic analyses of two mitochondrial and five nuclear markers, M. steenstrupi is classified in the predominantly free-living order Pleuronematida (Oligohymenophorea: Scuticociliatia). This order also encompasses other taxa isolated from molluscs and traditionally classified along with Myxophyllum in the order Thigmotrichida. The proper classifications of Myxophyllum was hampered by the dramatic remodelling of its oral apparatus (reduction of the paroral membrane and adoral organelles, formation of vestibular kineties), a transformation that was likely induced by its firm association with terrestrial gastropods. The present study also documents that various ciliate lineages independently became commensals or parasites of various aquatic and terrestrial molluscs.

Description of Three New Genera of Metopidae (Metopida, Ciliophora): Pileometopus gen. nov., Castula gen. nov., and Longitaenia gen. nov., with Notes on the Phylogeny and Cryptic Diversity of Metopid Ciliates

We report the discovery of three new species of freshwater metopid ciliates, Pileometopus lynni gen. et sp. nov., Castula flexibilis gen. et sp. nov., and Longitaenia australis gen. et sp. nov. Based on morphologic features and the 18S rRNA gene phylogeny, we transfer two known species of Metopus to the new genus Castula, as C. fusca (Kahl, 1927) comb. nov. and C. setosa (Kahl, 1927) comb. nov. and another known species is herein transferred to the new genus Longitaenia, as L. gibba (Kahl, 1927) comb. nov. Pileometopus is characterized by a turbinate body shape, a dorsal field of densely spaced dikinetids, a bipartite paroral membrane, and long caudal cilia. A distinctive morphologic feature of Castula species is long setae arising over the posterior third of the body (as opposed to a terminal tuft). Longitaenia spp. are characterized by an equatorial cytostome and long perizonal ciliary stripe relative to the cell length. Based on phylogenetic analyses of 18S rRNA gene sequences, we identify and briefly discuss strongly supported clades and intraspecific genetic polymorphism within the order Metopida.

Taxonomy and molecular phylogeny of three freshwater scuticociliates, with descriptions of one new genus and two new species (Protista, Ciliophora, Oligohymenophorea)

Three freshwater scuticociliates, Apouronema harbinensis gen. nov. spec. nov., Cyclidium vorax spec. nov., and C. glaucomaMüller, 1773, collected from rivers in Hulan District, Harbin, northeastern China, were investigated using morphological and phylogenetic criteria. Apouronema gen. nov., assigned to the family Uronematidae, is mainly distinguished from the other genera of the family by its paroral membrane extending anteriorly to the middle of membranelle 1. Apouronema harbinensis spec. nov. is defined by body size in vivo about 45–55 × 20–25 μm, buccal field about 70–80% of cell length; 12 or 13 somatic kineties; membranelle 1 having two rows, with 16–18 basal bodies in each kinety; membranelle 2 and membranelle 3 both having two rows each; scutica X-shaped with five pairs of basal bodies. Cyclidium vorax spec. nov. is characterized by the following features: body size 35–40 × 18–20 μm in vivo; 9 or 10 somatic kineties; membranelle 1 having two longitudinal rows, much shorter than M2; M2 triangle-shaped. The phylogenetic analyses show that: (1) Apouronema clustered in the Uronematidae clade, and grouped with genera Uronemita and Uronema; (2) Cyclidium vorax spec. nov. grouped with C. glaucoma and C. sinicum, which supports the assignment of the new species to the genus Cyclidium; (3) Cyclidium remains non-monophyletic with the addition of the new sequence.

The Taxonomy and Phylogeny of the Poorly known Heterotrich Ciliate Ampullofolliculina lageniformis Hadži, 1951 (Ciliophora: Heterotrichea).

Ampullofolliculina Hadži, 1951 is a monotypic genus, the only species being A.lageniformis which was first discovered in estuarine habitats in the U.S. and has never been redescribed. In the present study, we investigated the taxonomy and phylogeny of A.lageniformis Hadži, 1951 based on analyses of a population collected from a brackish wetland in Ningbo, China. The main characteristics of this species are as follows: trophont about 450-700μm long invivo with two short, rounded peristomial lobes of equal size; lorica flask-shaped, transparent and smooth with a short, wide neck at the base of which two transparent valves are asymmetrically inserted; cortex with brownish to reddish cortical granules and greenish pigment granules; about 80 somatic kineties evenly arranged; moniliform macronucleus with 4-8 ellipsoidal nodules; swarmer dark green, vermiform in shape, about 200-350μm long invivo, with about 60 adoral membranelles and 85 somatic kineties, no mouth nor paroral membrane. Phylogenetic analyses inferred from SSU rDNA sequences show that A.lageniformis is closely related to Folliculina and Eufolliculina which nest within a large clade that comprises five families, i.e. Stentoridae, Blepharismidae, Fabreidae, Maristentoridae, and Folliculinidae.

Morphology and Phylogenetic Position of Gruberia lanceolata (Gruber 1884) (Ciliophora, Heterotrichea) from Rio de Janeiro, Brazil.

Ciliates of the genus Gruberia are poorly studied. Consequently, most species lack detailed morphological descriptions, and all gene sequences in GenBank are not classified at the species level. In this study, a detailed morphological description of a population of G.lanceolata from Brazil is presented, based on live and protargol-stained organisms. We also present the 18S rRNA gene sequence and the phylogenetic position of this species. The primary characteristics of G.lanceolata from the Maricá Lagoon are as follows: an elongate fusiform body 280-870×40-160μm in size; rosy cortical granules; a peristome occupying approximately 1/3-1/2 of body length; an adoral zone comprising 115-330 membranelles; a paroral membrane in 35-50 fragments; and a moniliform macronucleus with 11-16 nodules. Based on our observations and data from pertinent literature, we suggest G.beninensis to be a junior synonym of G.lanceolata.

New contributions to Gruberia lanceolata (Gruber, 1884) Kahl, 1932 based on analyses of multiple populations and genes (Ciliophora, Heterotrichea, Gruberiidae)

Gruberia Kahl, 1932 is a species-poor genus comprising only seven named species. Most of these species have not been reinvestigated since the original reports. In the present work, we investigated the taxonomy and phylogeny of Gruberia lanceolata (Gruber, 1884) Kahl, 1932 based on analyses of morphology and multiple gene sequences from four South Korean populations. This species is mainly characterized by a well-developed peristome region, segmented paroral membrane, and moniliform macronucleus. Some morphological features were not stable among the four populations investigated, such as body shape and size, cell color, and the ratio of oral length to body length. However, our molecular analyses of four different genetic markers – three nuclear DNA markers (18S rDNA, ITS1-5.8S-ITS2 region, D1D2 of 28S rDNA) and one mitochondrial (mt) marker (CO1 gene) – indicated that all Korean populations examined were the same species. Based on our present findings and historic works, we propose that G. calkinsi, G. aculeata, and G. beninensis are junior synonyms of G. lanceolata.

One freshwater species of the genus Cyclidium, Cyclidiumsinicum spec. nov. (Protozoa; Ciliophora), with an improved diagnosis of the genus Cyclidium.

The morphology and infraciliature of one freshwater ciliate, Cyclidium sinicum spec. nov., isolated from a farmland pond in Harbin, northeastern China, was investigated using living observation and silver staining methods. Cyclidium sinicum spec. nov. could be distinguished by the following features: body approximately 20-25×10-15 µm in vivo; buccal field about 45-50 % of body length; 11 somatic kineties; somatic kinety n terminating sub-caudally; two macronuclei and one micronucleus; M1 almost as long as M2; M2 triangle-shaped. The genus Cyclidium is re-defined as follows: body outline usually oval or elliptical, ventral side concave, dorsal side convex; single caudal cilium; contractile vacuole posterior terminal; adoral membranelles usually not separated; paroral membrane 'L'-shaped, with anterior end terminating at the level of anterior end of M1; somatic kineties longitudinally arranged and continuous. Phylogenetic trees based on the SSU rDNA sequences showed that C. sinicum spec. nov. clusters with the type species, Cyclidiumglaucoma, with full support. Cyclidium is not monophyletic with members of the clade of Cyclidium+Protocyclidium+Ancistrum+Boveria.

A Huge Diversity of Metopids (Ciliophora, Armophorea) in Soil from the Murray River Floodplain, Australia. II. Morphology and Morphogenesis of Lepidometopus platycephalus nov. gen., nov. spec.

The morphology and morphogenesis of a new Australian metopid ciliate, Lepidometopus platycephalus nov. gen., nov. spec., were studied using live observation, various silver impregnation methods, scanning electron microscopy, and morphometry. The new genus is outstanding in having epicortical scales (lepidosomes) and a strongly flattened and distinctly projecting preoral dome. Diagnostic features of L. platycephalus include a small, reniform body carrying an elongated caudal cilium, about 11 ciliary rows, and an adoral zone composed of an average of 11 polykinetids. The morphogenesis of L. platycephalus matches data from other metopids in that (1) the body is drastically re-shaped, (2) the parental oral structures are reorganized but do not contribute to the daughter oral ciliature, (3) the opisthe’s adoral polykinetids originate pleurotelokinetally, (4) the opisthe’s paroral membrane is formed via re-arrangement of the posterior portion of the first two perizonal rows, and (5) the opisthe’s perizonal stripe is made by three parental perizonal rows and two dorsolateral ciliary rows. The morphogenetic data corroborate phylogenetic analyses in that caenomorphids are only superficially similar to metopids; metopids and clevelandellids are closely related; and litostomateans are the best candidates for a sister group of the metopid-clevelandellid assemblage within the SAL superclade.