Adoral Membranelles Research Articles

Bit by bit toward the diversity of metopids: Description of the genus Pidimetopus n. gen. (Ciliophora: Armophorea).

While metopids (Armophorea: Metopida) represent the most species-rich group of free-living anaerobic ciliates thriving in hypoxic environments, our understanding of their true diversity remains incomplete. Most metopid species are still characterized only morphologically. Particularly, the so-called IAC clade (named in the past after some of the taxa included, Idiometopus, Atopospira, and Clevelandellida), comprising free-living members as well as the endosymbiotic ones (order Clevelandellida), is in serious need of revision. In our study, we establish a new free-living genus in the IAC clade, Pidimetopus n. gen., with descriptions of two new species, P. nanus n. sp., and P. permonicus n. sp., using up-to-date molecular and morphologic methods. The genus is characterized by small cells (up to 75 μm long), not more than 10 adoral membranelles and eight somatic kineties, and usually, four long caudal cilia that can stiffen. In addition to morphologic and molecular characterizations, we also conducted a statistical morphotype analysis of the polymorphic species P. nanus n. sp. We discuss the relevance of the earlier morphologically described species Metopus minor as a putative collective taxon for several small metopids less than 50 μm long.

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.

Morphology, morphogenesis and molecular phylogeny of a new soil ciliate, Lamtostyla paravitiphila nov. spec. (Ciliophora, Hypotrichia)

The morphology and morphogenesis of Lamtostyla paravitiphila nov. spec., a novel soil hypotrichous ciliate collected from eastern China, were investigated based on live observations and protargol-stained specimens. The new species is morphologically characterized as follows: seven to twelve macronuclear nodules, cortical granules absent, 19–26 adoral membranelles, three or four frontoventral cirri, the amphisiellid median cirral row extends to about mid-body and composed of 12–18 cirri, two or three transverse cirri, 27–39 left and 30–41 right marginal cirri, three almost bipolar dorsal kineties. Morphogenetically, it is characterized by the initial formation of six frontal-ventral-transverse cirral anlagen as primary primordia. Notably, the amphisiellid median cirral row and the posterior frontoventral cirrus (or cirri) contribute to the development of the frontal-ventral-transverse cirral anlagen, while the buccal cirrus may not participate in this process. Phylogenetic analyses based on small subunit ribosomal DNA sequence data indicate that the Lamtostyla species with available molecular data do not form a monophyletic group.

Relating ciliary propulsion morphology and flow to particle acquisition in marine planktonic ciliates II: the oligotrich ciliate Strombidium capitatum

Abstract The marine oligotrich ciliate Strombidium capitatum is a cruise-feeder, relying on ciliary motion and propulsion flow to individually detect and capture particles. High-speed, high-magnification digital imaging revealed that the cell swims forward by sweeping its anterior adoral membranelles (AAMs) backward, achieving a mean path-averaged speed of U = 1.7 mm s−1 (31 cell-lengths per second). Particle detection occurs through either hydrodynamic signal perception or ciliary contact perception, with a mean reaction distance of R = 20.4 μm. While executing a ciliary reversal of AAMs to handle and capture a perceived particle, the cell coordinates the ciliary motion of ventral adoral membranelles (VAMs, the “lapel”) with the ciliary reversal of AAMs (the “collar”), causing a sudden halt of cell motion, thereby functioning as a motion “brake” that is crucial for effective particle capture. The encounter rate with small prey particles is calculated using πR2U (~8.0 μL h−1, equivalent to ~ 3.5 × 106 cell volumes per day). Based on hydrodynamic modeling results, it is hypothesized that spatial structures of the flow velocity vector and acceleration fields in front of the swimming cell are essential for pushing an embedded particle forward, creating a strong enough slip velocity and hydrodynamic signal for prey perception, even for a neutrally buoyant small particle.

Morphology and molecular phylogeny of two hypotrichous ciliates (Ciliophora, Spirotrichea) from South Korea, including Hemiurosomoida koreana n. sp.

The living morphology, infraciliature, and molecular phylogeny of a new soil ciliate, Hemiurosomoida koreana n. sp., discovered in a sample collected from a mountain in the northeast of South Korea, were investigated. The new species possesses the characteristics of the genus Hemiurosomoida, i.e., a reduced number of frontal-ventral-transverse cirri, three dorsal kineties of which kineties 1 and 2 each bears a caudal cirrus, and a single dorsomarginal kinety. It is distinguishable from congeners and other similar species by at least one distinct qualitative or quantitative character including the body size, the presence and arrangement of cortical granules, the number of adoral membranelles, marginal cirri, and dorsal dikinetids, or by the arrangement of transverse cirri. Phylogenetic analyses based on 18S rRNA gene sequences also support the assignment of the new species to the non-monophyletic genus Hemiurosomoida. In addition, the living morphology, infraciliature, and the 18S rRNA gene sequence of a Korean population of Nothoholosticha flava were studied.

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.

Trichite features contribute to the revision of the genus Strombidium (Alveolata, Ciliophora, Spirotricha).

Strombidium is a species-rich genus of oligotrichid ciliates mainly inhabiting the marine pelagial. In molecular phylogenies, the genus emerged as non-monophyletic, and cladistic analyses suggest that it is largely characterized by plesiomorphies. A reliable split of the genus and the establishment of new genera necessitate, however, support by novel morphological and/or ultrastructural features. In the present study, the arrangement and ultrastructure of trichites are proposed as taxonomically relevant characters. Strombidium biarmatum Agatha et al., 2005 differs in the trichite pattern from the type species Strombidium sulcatum and most congeners. Aside from the trichites inserting anteriorly to the girdle kinety and generating the typical funnel-shaped complex in the posterior cell portion, the species displays additional trichites between the adoral membranelles even visible in live cells. Here, this exceptional trichite arrangement is detailed based on transmission electron microscopic investigations. In molecular phylogenies, S. biarmatum forms a monophylum with two congeners sharing its trichite arrangement. Therefore, the strombidiid genus Heteropilum nov. gen. is established with S. biarmatum as type species to also include H. paracapitatum (Song et al., 2015) nov. comb. and H. basimorphum (Martin & Montagnes, 1993) nov. comb. Further differences discovered in the trichite ultrastructure support the organelles' taxonomic significance.

Morphology and phylogenetic position of three anaerobic ciliates from the classes Odontostomatea and Muranotrichea (Ciliophora).

The diversity of the classes Odontostomatea and Muranotrichea, which contain solely obligate anaerobes, is poorly understood. We studied two populations of Mylestoma sp., one of Saprodinium dentatum (Odontostomatea), two of Muranothrix felix sp. nov., and one of Muranothrix sp. (Muranotrichea) employing live observation, protargol impregnation, scanning electron microscopy, and 18S rRNA gene sequencing. Conspecificity of Mylestoma sp., described here, with a previously described species of this genus cannot be excluded since no species have been studied with modern methods. Phylogenetically, the genus Mylestoma is closely related to the odontostomatid Discomorphella pedroeneasi, although the phylogenetic position of class Odontostomatea itself remains unresolved. The newly described muranotrichean species, Muranothrix felix sp. nov., is morphologically similar to M. gubernata but can be distinguished by its fewer macronuclear nodules and fewer adoral membranelles; moreover, it is clearly distinguished from M. gubernata by its 18S rRNA gene sequence. Another population, designated here as Muranothrix sp., most likely represents a separate species.

Strongylidium koreanum n. sp. (Protozoa: Ciliophora), a new soil species from South Korea.

Strongylidium koreanum n. sp., a new soil ciliate from Jeju Island, South Korea, is described based on live observations, protargol impregnation, and molecular analysis of the 18S rRNA gene sequence. It is characterized by the following morphological features: cell outline more or less fusiform, posterior end broader than anterior end; grayish under low magnification; cortical granules absent; 23-32 adoral membranelles; three enlarged frontal cirri; buccal cirrus and postoral ventral cirrus present; 27-42 left and 15-28 right ventral cirri; 23-36 left and 30-46 right marginal cirri; three dorsal kineties; three caudal cirri; and two macronuclear nodules with two or three micronuclei. Phylogenetic analyses show that Strongylidium is monophyletic.

Ultrastructure of Diophrys appendiculata and new systematic consideration of the euplotid family Uronychiidae (Protista, Ciliophora).

The ultrastructure of ciliates carries important cytological, taxonomical, and evolutionary signals for these single-celled eukaryotic organisms. However, little ultrastructural data have been accumulated for most ciliate groups with systematic problems. In the present work, a well-known marine uronychiid, Diophrys appendiculata, was investigated using electron microscopy and a comparison with, and a discussion considering, phylogenetic analyses were made. The new findings primarily show that: (i) this species lacks the typical alveolar plate, bears cortical ampule-like extrusomes, and has microtubular triads in the dorsal pellicle, and thus exhibits some ultrastructural features in common with most of its previously studied congeners; (ii) each adoral membranelle before the level of frontal cirrus II/2 contains three rows of kinetosomes and each membranelle after the level of frontal cirrus II/2 contains four rows, which might be related with morphogenesis and could be considered as a distinctive character of Diophrys; (iii) some structural details of the buccal field, such as the extra-pellicular fibrils, pellicle, pharyngeal disks and microtubular sheet, were documented. In addition, based on the ultrastructural comparison of representatives, we discuss the differentiation between the subfamilies Diophryinae and Uronychiinae. A hypothetical systematic relationship of members in the order Euplotida based on a wide range of data is also provided.

Morphology and Morphogenesis of Pseudourostyla levis (Ciliophora, Hypotrichia) from River Yamuna, Delhi, India.

The ciliated protozoa, Pseudourostyla levis was collected from river Yamuna in Delhi (India). Morphometric characterization and cortical morphogenesis during the division cycle was investigated. The ciliate Pseudourostyla measures 274µm × 80 µm in protargol stained preparations. Cortical structures include buccal, frontal and somatic ciliature. The main morphogenetic events during division cycle in P. levisincludes, partial retention of parental adoral membranelles, dedifferentiation of parental midventral cirri to form frontoventral complex and formation of marginal rows on each side from a common marginal primordium. In the present study, the morphometric comparison of the present isolate with congeners is also presented.

Diophrys pauciciliata n. sp. (Ciliophora, Euplotida), a new marine spirotrich ciliate with highly static morphometrics

A new marine ciliate was discovered from intertidal mudflat from South Korea. Based on the observation of living cells, protargol-impregnated specimens and SEM, we describe it as new species belonging to the genus Diophrys Dujardin, 1841, namely Diophrys pauciciliata n. sp. Its nuclear small subunit ribosomal RNA gene also supports the taxonomic assignment. Diophrys pauciciliata n. sp. is one of the smallest species in the genus. Besides, the bipartite adoral zone and oligomerized dorsal bristles easily discriminate the new species from other congeners. Interestingly, D. pauciciliata n. sp. showed highly static states in all morphometrics (<8.5 % of coefficient of variation) and in particular, the number of frontal adoral membranelles and the counts of dorsal ciliature (e.g., number of dorsal kineties and numbers of dikinetids within each kinety) are invariable. Considering morphometric data of previous studies in the genus Diophrys, the narrow range of character states is unexpected because cell length is usually positively correlated with the numbers of adoral membranelles and dorsal dikinetids.

Morphology and molecular phylogeny of Keronopsis koreana n. sp. (Ciliophora, Hypotricha), a new soil species from South Korea

The morphology and molecular phylogeny of a new soil ciliate, Keronopsis koreana n. sp., which was discovered in soil from Jindo-gun, South Korea, were investigated. The new species is characterized by the following morphological features: cell more or less ellipsoid or ovoid, both ends broadly rounded; cortical granules irregularly distributed, colorless, 1 µm across; 35–54 adoral membranelles; 12–23 cirri in frontal corona; two frontal and two frontoventral cirral rows; two or three buccal cirri; 23–38 left and 22–37 right marginal cirri; three bipolar dorsal kineties; and two macronuclear nodules and three to five micronuclei. Phylogenetic analyses show that Keronopsis koreana n. sp. is placed within a clade containing K. helluo (type species of Keronopsis) and species belonging to Paraholosticha, Uroleptoides, Parabistichella, Orthoamphisiella, Lamtostyla, and Bistichella in non-dorsomarginalian hypotrichs. Also, the sequences belonging to the Keronopsidae did not cluster together. In addition, a new combination is provided: Parabistichella tasmaniensis (Blatterer and Foissner, 1988) n. comb. (original combination: Keronopsis tasmaniensis).

Two hypotrichs (Ciliophora, Hypotricha) from China: morphology and SSU rDNA sequence of Holosticha aestuarina nov. spec. and H. muuiensis Kim et al., 2017

The morphology, four ontogenetic stages, and the phylogenetic relationships based on the small subunit RNA-gene of Holosticha aestuarina nov. spec. from the Yangtze River Estuary (Shanghai) were investigated. The new species differs from the congeners by the following combination of features: body size 160–196 × 27–36 μm in vivo, a vesicular structure containing one or two bean-shaped opaque particles, 24–32 adoral membranelles, 16–34 midventral pairs, 17–26 left marginal cirri, 22–32 right marginal cirri, six or seven transverse cirri, four dorsal kineties, and usually two macronuclear nodules. The anlage for the left marginal row of the proter very likely originates de novo (anterior portion) and from parental cirri (posterior portion). Further, we provide a morphological description of a Chinese population of H. muuiensis Kim et al., 2017, originally discovered in South Korea, indicating that it is confined to the Eastern Asia region. Its SSU rDNA sequence is identical with that of the type population and forms a clade with the available Holosticha sequences. By contrast, H. aestuarina nov. spec. is sister to the Holosticha + Uncinata clade, indicating that Uncinata is a subgenus of Holosticha.

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.

Taxonomy and SSU rRNA gene-based phylogeny of two new Euplotes species from China: E. chongmingensis n. sp. and E. paramieti n. sp. (Protista, Ciliophora)

BackgroundThe genus Euplotes Ehrenberg, 1830, one of the most complicated and confused taxa, contains about 160 nominal species. It was once proposed to be divided into four genera, two of which were proved to be non-monophyletic. At least 19 new species have been discovered in the past decade, implying that there is a large undiscovered diversity of this genus.ResultsThe morphology of two new freshwater euplotid ciliates, Euplotes chongmingensis n. sp. and E. paramieti n. sp., isolated from Shanghai, China, were investigated using live observations, protargol staining, and Chatton-Lwoff silver staining method. Euplotes chongmingensis is characterized by its small size (40–50 × 25–35 μm), about 24 adoral membranelles, 10 frontoventral cirri, two marginal and two caudal cirri, eight dorsolateral kineties with 11–16 dikinetids in the mid-dorsolateral kinety and a double type of silverline system. Euplotes paramieti n. sp. is 180–220 × 110–155 μm in vivo and strongly resembles E. amieti but having a difference of 57 bp in their SSU rRNA gene sequences. Phylogenetic analyses based on SSU rRNA gene sequence data were used to determine the systematic positions of these new taxa.ConclusionsThe description of two new freshwater taxa and their SSU rRNA gene sequences improve knowledge of biodiversity and enrich the database of euplotids. Furthermore, it offers a reliable reference for environmental monitoring and resource investigations.

Taxonomy and Phylogeny of Two Spirotrichean Ciliates (Alveolata, Protista, and Ciliophora) From a Plateau Saline–Alkaline Lake in Tibet, China, Including Description of a New Species

There are few researches concerning ciliates from “extreme” environments such as plateau salt lakes; nevertheless, such a faunistic survey is of great significance for understanding the diversity and biogeography of these microeukaryotes. In this study, two ciliates, namely, Hemiurosomoida tibetensis sp. nov. and Euplotes euryhalinusValbonesi and Luporini, 1990, isolated from Kyêbxang Co, a Tibet Plateau saline-alkaline lake, China, were investigated based on observations of live and silver-stained specimens, and 18S rDNA sequences were analyzed. The new species can be characterized by having a size of 60–95 × 20–40 μm in vivo, an elongate elliptical body, colorless cortical granules mainly grouped around cirri and dorsal bristles, about 21 adoral membranelles, 20 cirri on each marginal row, and two macronuclear nodules and two micronuclei. The Chinese population of E. euryhalinus corresponds well with the original one. Phylogenetic analyses show that Hemiurosomoida is non-monophyletic, H. tibetensis sp. nov. has a closer relationship with H. longa within this genus, and that Euplotes euryhalinus presents a wide geographic distribution. This study first reveals the ciliature, morphogenesis, and rRNA gene sequence data for H. tibetensis sp. nov. and the Chinese population of E. euryhalinus, which thus increases our knowledge about the diversity of ciliates at high altitudes.

A New Protocruzia Species (Ciliophora: Protocruziida) Isolated From the Mariana Trench Area.

A new species of Protocruzia, isolated from the deep-sea Pacific Ocean (>3,000-m depth) in the vicinity of the Mariana Trench, is described based on morphological and molecular data. The systematic status of the ciliate genus Protocruzia has long been highly ambiguous, and Protocruzia species have been assigned to an independent class until recently. In the present study, we described Protocruzia marianaensis sp. n. as a small (25–32 × 14–17 μm in vivo) drop-shaped ciliate, with longitudinal furrows along the ciliary rows on the right side, six adoral membranelles, eight somatic kineties, and one macronucleus comprising 7–11 nuclear globules. Phylogenetic analyses inferred from small subunit rRNA gene sequences revealed that seven Protocruzia species in the phylogenetic tree formed a fully supported clade representing an independent class. Protocruzia marianaensis sp. n. was established to be most closely related to Protocruzia adhaerens, with a sequence similarity of 96.64%, and was found to be able to survive at both atmospheric pressure and hydrostatic pressure of 320 bar, thereby indicating effective barotolerance.

Morphology, Morphogenesis, and Molecular Phylogeny of a New Freshwater Ciliate, Quadristicha subtropica n. sp. (Ciliophora, Hypotrichia).

The morphology and the regulation of cortical pattern associated with the cell size, division, and phylogenetic position of a new hypotrichous ciliate, Quadristicha subtropica n. sp. collected from a freshwater pond in southern China, were investigated. Quadristicha subtropica n. sp. is characterized as follows: size in vivo 60–115 μm × 25–45 μm; 19–21 adoral membranelles; buccal cirrus near anterior end of endoral and paroral; cirrus IV/3 at about level of buccal vertex; right marginal row begins ahead of buccal vertex; 11–16 right and 12–19 left marginal cirri; and dorsal cilia about 5 μm long. The basic morphogenetic process in Q. subtropica n. sp. is consistent with that of the type species, Quadristicha setigera. Phylogenetic analyses based on small subunit ribosomal DNA sequence data reveal that the systematic position of Q. subtropica n. sp. is rather unstable with low support values across the tree and the genus Quadristicha is not monophyletic.

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.