Functional Ecology of Two Contrasting Freshwater Ciliated Protists in Relation to Temperature.

We conducted microcosm experiments with two contrasting freshwater ciliates on functional traits (FTs) related to their growth rate (numerical response, NR) and ingestion rate (functional response, FR) over a range of ecologically relevant temperatures. Histiobalantium bodamicum and Vorticella natans are common planktonic ciliates but their abundance, swimming behavior, and temperature tolerance are different. In contrast to most sessile peritrich species, the motile V. natans is not strictly bacterivorous but also voraciously feeds upon small algae. We observed three main alterations in the shape of NR of both species with temperature, that is, change in the maximum growth rate, in the initial slope and in the threshold food level needed to sustain the population. Similarly, maximum ingestion rate, gross growth efficiency (GGE), and cell size varied with temperature and species. These findings caution against generalizing ciliate performance in relation to the ongoing global warming. Our results suggest that V. natans is the superior competitor to H. bodamicum in terms of temperature tolerance and bottom-up control. However, the abundance of V. natans is usually low compared to H. bodamicum and other common freshwater ciliates, suggesting that V. natans is more strongly top-down controlled via predation than H. bodamicum. The taxonomic position of V. natans has been debated. Therefore, to confirm species and genus affiliation of our study objects, we sequenced their small subunit ribosomal RNA (SSU rDNA) gene.

The insights into the systematic relationship of Gastrostyla-affinitive genera, with report on a new saline soil ciliate genus and new species (Protozoa, Ciliophora).

BACKGROUND: Hypotrichia are a group with the most complex morphology and morphogenesis within the ciliated protists. The classification of Gastrostyla-like species, a taxonomically difficult group of hypotrichs with a common ventral cirral pattern but various dorsal and ontogenetic patterns, is poorly understood. Hence, systematic relationships within this group and with other taxa in the subclass Hypotrichia remain unresolved. RESULTS: 18S rRNA gene sequence of a new Gastrostyla-like taxon was obtained. Phylogenetic analyses based on the 18S rRNA gene sequences indicate that this ciliate represents a new genus that is closely related to Heterourosomoida and Kleinstyla within the oxytrichid clade of the Hypotrichia. However, the position of this cluster remains unresolved. All three genera deviate from the typical oxytrichids by their incomplete (or lack of) dorsal kinety fragmentation during morphogenesis. Morphology and morphogenesis of this newly discovered form, Heterogastrostyla salina nov. gen., nov. spec., are described. Heterogastrostyla nov. gen., is characterised as follows: more than 18 fronto-ventral-transverse cirri, cirral anlagen V and VI develop pretransverse cirri, and dorsal ciliature in Urosomoida-like pattern. CONCLUSIONS: Similar to the CEUU-hypothesis about convergent evolution of urostylids and uroleptids, we speculate that the shared ventral cirral patterns of Gastrostyla-like taxa might have resulted from convergent evolution.

Morphology and Morphogenesis of a Novel Saline Soil Hypotrichous Ciliate, Gonostomum sinicum nov. spec. (Ciliophora, Hypotrichia, Gonostomatidae), Including a Report on the Small Subunit rDNA Sequence.

Morphology, cirral pattern, and morphogenesis of the new saline soil hypotrich, Gonostomum sinicum nov. spec. collected from Longfeng Wetland in Daqing, north China, were studied, using detailed live observations and protargol-stained specimens. The new species is characterized as follows: (i) a size in vivo of 100-125 × 30-40 µm, (ii) colorless cortical granules, 0.5 µm across, arranged in short rows, (iii) an adoral zone composed of 28-33 membranelles, (iv) three or four frontoventral rows, one of which extends onto the postoral area, (v) left and right marginal rows composed of 18-27 and 21-35, cirri, respectively, and (vi) usually two transverse cirri. Morphogenesis is as usual for the genus Gonostomum, i.e. the cirral primordia II-VI are primary primordia which split into two sets for proter and opisthe in division middle stages, except for anlage I which develops independently. However, the number of frontoventral transverse anlagen is either five or six not only in different individuals but even in proter and opisthe of the same divider. The phylogenetic analyses based on SSU rDNA sequences showed that the genus Gonostomum is nonmonophyletic, indicating that the patterns of cirri and dorsal kineties are homoplasious characters. The new species G. sinicum nov. spec. is perhaps closely related to Cotterillia bromelicola and two congeners.

Reconsideration of the 'well-known' hypotrichous ciliate Pleurotricha curdsi (Shi et al., 2002) Gupta et al., 2003 (Ciliophora, Sporadotrichida), with notes on its morphology, morphogenesis and molecular phylogeny.

The oxytrichid species Pleurotricha curdsi (Shi et al., 2002) Gupta et al., 2003, isolated from a tributary of the Yangtze River in the Mudong district of Chongqing, southern China, was reinvestigated with emphasis on its morphology, morphogenesis and small-subunit (SSU) rDNA-based phylogeny. Compared with three previously described populations, the Mudong population of P. curdsi is characterized by its large body size, 170-295 × 65-110 µm in vivo, and by having a variable number of right marginal rows, either two or three. Likewise, the number of right marginal rows anlagen (RMA) is also variable, i.e. usually two, but sometimes several small extra anlagen that give rise to the formation of the third row, are present to the left of the RMAs. We posit that the Mudong population is an intermediate form between the three previously described populations. Phylogenetic analyses based on the SSU rDNA sequence data show that all populations of P. curdsi cluster with the type species of the genus, Pleurotricha lanceolata, in a clade nested within the Oxytrichidae.

Community assembly and co-occurrence network complexity of interstitial microbial communities in the Arctic (investigation of ciliates in the White Sea intertidal zone).

Arctic coastal ecosystems play a major role in global environmental system and have been altered significantly by climate changes. To better understanding the response of marine coastal ecosystems towards rapid Arctic climate changes, we examined the variation in diversity and community structure and provided insights into the co-occurrence network and community assembly of interstitial ciliates in the Kandalaksha Gulf of the White Sea from 2009 to 2019. Co-occurrence networks analysis indicated considerably high ration of positive correlations within a community that indicated low competition between interstitial ciliate species. Furthermore, we found that contribution of stochastic processes to the ciliate community assembly was insignificant. Compare with earlier data from the same ecosystem obtained in 1980s-1990s, the role of competitive factors is decreasing, and communities are becoming more spatially and temporally homogeneous. This community simplification is likely due to the response of the entire intertidal ecosystem to global climate change in Arctic.

Top-down control of planktonic ciliates by microcrustacean predators is stronger in lakes than in the ocean.

Planktonic ciliates are major components of pelagic food webs in both marine and freshwaters. Their population dynamics are controlled 'bottom-up' by prey availability and 'top-down' by microcrustacean predators. In oceans, copepods are the main ciliate predators while in lakes cladocerans are the typical predators. The efficacy by which these functionally different predators control ciliate population dynamics is debated. We, therefore, investigated experimentally the grazing of three microcrustacean predators with different feeding modes on five freshwater ciliates. We then performed a meta-analysis to assess if our findings can be generalised for aquatic ecosystems. We hypothesized that top-down control is stronger in lakes than in the ocean. We find that: (i) average ingestion rates of marine and freshwater microcrustaceans do not differ; (ii) clearance rates of freshwater cladocerans decrease with ciliate size but increase with ciliate size in freshwater copepods; (iii) clearance rates of the marine microcrustaceans is unrelated to ciliate cell size. These findings have implications for the functioning of freshwater and marine food webs: (i) the ciliate-microcrustacean link is stronger in lakes than in the ocean, and (ii) globally top-down control of ciliates is unlikely in the ocean.

Doublets of the unicellular organism Euplotes vannus (Alveolata, Ciliophora, Euplotida): the morphogenetic patterns of the ciliary and nuclear apparatuses associated with cell division.

Ciliated protists are one of the most diverse and highly differentiated group among unicellular organisms. Doublets occur in ciliates when two cells fuse into a single individual. Doublets contain two major cellular components (either cell in a doublet) and have traditionally been considered as developmental anomalies. Nevertheless, doublets can divide or even conjugate effectively, which may represent dispersal forms of the life stages. In addition, morphogenesis, as an important process in the life cycle, will provide important insights into the complex differentiation mechanism and various physiological phenomena. However, morphogenetic studies focusing on doublets of ciliates are very limited, which has become an obstacle to understand their complete life history. Here we isolated a doublet strain from the marine species Euplotes vannus (Müller, 1786) Diesing, 1850 and investigated its morphogenetic events during asexual reproduction. Our results indicate that: (1) the opisthe's oral primordium develops de novo beneath the cortex; (2) the frontoventral and transverse cirral anlagen, cirrus I/1, and marginal anlagen in both dividers develop de novo separately; (3) the dorsal kinety anlagen, the three rightmost ones of which produce three caudal cirri for the proter, occur within the parental structures in the mid-body region; (4) the opisthe acquires two caudal cirri, one from the end of each two rightmost kineties; and (5) there are two macronuclei and one micronucleus in the doublet and they divide amitotically and mitotically, respectively. Finally, we speculate that this special differentiation may be an adaptive form to adverse environments.

Hypotrichidium tisiae (Gelei, 1929) Gelei, 1954: a unique hypotrichid ciliate having a highly specialized developmental pattern during binary division.

In sharp contrast to their pelagic relatives, the oligotrichs, the overwhelming majority of hypotrich ciliates inhabit the benthos. Only a few species, including those of the genus Hypotrichidium Ilowaisky, 1921, have adapted to a planktonic lifestyle. The ontogenetic mode of the highly differentiated ciliate, Hypotrichidium tisiae (Gelei, 1929) Gelei, 1954, is unknown. In this study, the interphase morphology and the ontogenetic process of this species are investigated. Accordingly, the previously unidentified ciliary pattern of Hypotrichidium is redefined. The main morphogenetic features are as follows: (1) The parental adoral zone of membranelles is inherited completely by the proter and the oral primordium of the opisthe arises in a deep pouch. (2) Five frontoventral cirral anlagen (FVA) are formed: FVA I contributes to the single frontal cirrus, FVA II-IV generate three frontoventral cirral rows, FVA V migrates and forms postoral ventral cirri. (3) All marginal cirral row anlagen develop de novo: each of the two left anlagen forms a single cirral row, while the single right anlage fragments into anterior and posterior parts. (4) Two dorsal kinety anlagen occur de novo, with the right one fragmenting to form kineties 2 and 3. (5) Two long caudal cirral rows are formed at the ends of dorsal kineties 1 and 3. On the basis of the morphogenetic features and phylogenetic analyses, the assignment of Hypotrichidium to the family Spirofilidae Gelei, 1929 within Postoralida is supported. The establishment of separate families for the slender "tubicolous" spirofilids and the highly helical spirofilids is also validated. Supplementary Information: The online version contains supplementary material available at 10.1007/s42995-022-00148-9.

Timing and characteristics of nuclear events during conjugation and genomic exclusion in Paramecium multimicronucleatum.

Ciliated protists are ideal material for studying the origin and evolution of sex, because of their nuclear dimorphism (containing both germline micronucleus and somatic macronucleus in the same cytoplasm), special sexual processes (conjugation and autogamy), and high diversity of mating-type systems. However, the study of sexual process is limited to only a few species, due to the difficulties in inducing or observing conjugation. In the present study, we investigate the conjugation process in Paramecium multimicronucleatum: (1) of the three prezygotic divisions, all micronuclei undergo the first two divisions (meiosis I, II), while a variable number of nuclei undergo the third division (mitosis); (2) the synkaryon divides three times after fertilization, giving rise to eight products that differentiate into four macronuclear anlagen and four micronuclei; (3) cells restore the vegetative stage after two successive cell fissions during which the macronuclear anlagen are distributed into daughter cells without division, while micronuclei divide mitotically; (4) the parental macronucleus begins to fragment following the first meiotic division and finally degenerates completely; (5) the entire process takes about 110 h, of which about 85 h are required for macronuclear development. In addition, we describe for the first time the process of genomic exclusion occurring between amicronucleate and micronucleate cells of P. multimicronucleatum, during which the micronucleate cell contributes a pronucleus to the amicronucleate cell, resulting in both exconjugants being homozygotes. These results provide new insights into the diversity of sexual processes and lay an important cytological basis for future in-depth studies of mating systems in ciliates.

Container volume may affect growth rates of ciliates and clearance rates of their microcrustacean predators in microcosm experiments.

We studied the effect of volume in small containers (microcosms) on five common planktonic freshwater ciliates and three zooplankton species, namely Daphnia sp., the calanoid copepod Eudiaptomus sp., and the cyclopoid copepod Cyclops sp. We measured ciliate specific growth rates and their loss rates due to microcrustacean predation in short-term experiments. We hypothesized that container volume ranging from 10 to 200 mL would not affect the activity of our prey and predator species. We found that the response to volume was species-specific; growth rates of three ciliate species were sensitive to volume. However, the volume effect was not unequivocal because different timing of the microcosm experiments (block effects) may have caused random bias due to varying morphological and/or physiological conditions of the ciliates and their predators. For predator clearance rate, the volume effect was insignificant in the filter-feeding Daphnia and Eudiaptomus but was significant for the predatory copepod Cyclops, which was hampered in the smallest experimental containers. Total crustacean clearance rates averaged over all treatments appeared unaffected by predator species, while ciliate species significantly affected the results. Our growth and clearance rates are close to previous findings with the same or similar planktonic prey and predator species.

Does the Gonostomum-patterned oral apparatus in hypotrichia carry a phylogenetic signal? Evidence from morphological and molecular data based on extended taxon sampling using three nuclear genes (Ciliophora, Spirotrichea).

The classification of hypotrichs based on the gonostomatid oral structure is widely accepted, but the phylogenetic signal of this character is unknown. Here, we infer the species phylogeny of those gonostomatids for which molecular data are available, plus 26 new sequences of SSU-rDNA, ITS1-5.8S-ITS2 and LSU-rDNA genes. The results indicate that: (i) the endoral is more phylogenetically informative than the paroral; (ii) the structure of the endoral and the Gonostomum-pattern adoral zone of membranelles are plesiomorphies for the hypotrichs sensu stricto; (iii) the group of species possessing these features is monophyletic in all our phylogenetic analyses, except that for the SSU-rDNA; (iv) Schmidingerotrichidae is monophyletic in all trees, suggesting that it is a well-defined family; (v) the Gonostomatidae is polyphyletic in the SSU-rDNA and ITS1-5.8S-ITS2 trees, with Gonostomum, Cladotricha, Cotterillia, Metagonostomum, Paragonostomum and Wallackia distributed among separate clades, but monophyletic in the LSU-rDNA and concatenated trees; (vi) higher hypotrich taxa such as core urostyloids and core sporadotrichids/stichotrichids might have evolved from species that possessed a gonostomatid oral apparatus.

Morphology, cell-division, and phylogeny of Schmidingerothrix elongata spec. nov. (Ciliophora, Hypotricha), and brief guide to hypotrichs with Gonostomum-like oral apparatus.

The hypotrich Schmidingerothrix elongata spec. nov., discovered in saline (20‰) soil of the Longfeng Wetland, Daqing, northern China, was studied using live observation and protargol impregnation. It is characterized, inter alia, by colorless cortical granules arranged in short rows, three frontoventral cirral rows with the rightmost extending far posteriorly, and 4-8, usually six macronuclear nodules. Cell division proceeds as in congeners and confirms the lack of dorsal ciliature. In phylogenetic analyses based on SSU rDNA, S. elongata is sister of S. salinarum+Paracladotricha salina. A re-investigation of the type slides of P. salina, type of Paracladotricha, revealed a misobservation in the original description. Since P. salina lacks, like Schmidingerothrix spp., a dorsal ciliature, Paracladotricha becomes a junior, subjective synonym of Schmidingerothrix with S. salina comb. nov. as fourth species. A review of the phylogenetic analyses dealing with Schmidingerothrix shows that its position is variable. However, together with the gonostomatid oral apparatus it can be hypothesized that Schmidingerothrix is a member of the Gonostomatidae or a close relative. A list of genera (14) and species (58) which have - like Schmidingerothrix - a gonostomatid oral apparatus, as well as a key to these genera are provided.

The diverse morphogenetic patterns in spirotrichs and philasterids: Researches based on five-year-projects supported by IRCN-BC and NSFC.

In the five years 2012-2016, the ciliate research group at Ocean University of China and their collaborators have performed several investigations on the morphogenesis of ciliated protists during binary division. Multiple samples were collected from 17 cities and cortical development studied in 42 species belonging to 32 genera and 13 families (Amphisiellidae, Euplotidae, Kahliellidae, Oxytrichidae, Philasteridae, Pseudokeronopsidae, Pseudourostylidae, Schmidingerotrichidae, Spirofilidae, Strobilidiidae, Uroleptidae, Uronychiidae and Urostylidae). Among these, 12 genera were investigated morphogenetically for the first time, revealing some unusual pattern formations and allowing four new genera to be established: Heterokeronopsis, Apobakuella, Parabistichella and Apoholosticha. The objective of this review is to: 1) summarize the morphogenetic studies supported by the IRCN-BC and NSFC projects during these five years; 2) summarize the patterns of development and document deviations from normal morphogenetic events within a group; 3) discuss how studies on morphogenesis have helped to advance understanding in the three dimensions of biodiversity, i.e. taxonomy, genetics and function; and 4) suggest potential future directions for the morphogenetic study of ciliated protists.

Redescription of two little-known urostyloid ciliates, Anteholosticharandani (Grolière, 1975) Berger, 2003 and A. antecirrata Berger, 2006 (Ciliophora, Urostylida).

The morphology of two little-known urostyloid ciliates, Anteholosticha randani (Grolière, 1975) Berger, 2003 and A. antecirrataBerger, 2006, collected from freshwater biotopes in southern China, was studied based on live observations and protargol staining. Anteholosticha randani is characterized by its bipartite adoral zone and short, longitudinally aligned undulating membranes. One early stage of reorganization/morphogenesis, one early-middle stage of reorganization and one middle stage of morphogenesis are also reported. Anteholosticha antecirrata is characterized by its large body size in vivo (200-400 × 40-80 µm), a row of buccal cirri and conspicuous, yellow-green cortical granules. Phylogenetic analyses based on SSU rDNA sequence data reveal that A. antecirrata may share a most recent common ancestor with Urostyla grandis and Bakuella granulifera, whereas A. randani branches independently and is sister to a large clade that includes Pseudourostyla, Pseudokeronopsis, Caudiholosticha and several species of Anteholosticha.

Morphology, morphogenesis and molecular phylogeny of a novel saline soil ciliate, Lamtostyla salina n. sp. (Ciliophora, Hypotricha).

The morphology and morphogenesis of a new saline soil hypotrich, Lamtostyla salina n. sp., collected from Longfeng Wetland in Daqing, north China, were studied based on live observations and protargol stained specimens. The new species is characterized as follows: body very flexible but not contractile, lanceolate with anterior end broadly rounded, widest at about 1/3 of body length, posterior end narrowly rounded; cortical granules colourless and scattered; amphisiellid median cirral row ends ahead of mid-body, composed of 5-13 cirri; 4-20 frontoventral cirri arranged in 2-4 rows; three frontal, one buccal and 2-6 transverse cirri; usually one left and one right marginal row, composed of 17-51 and 20-51 cirri respectively; usually two dorsal kineties; two macronuclear nodules and two micronuclei. Morphogenesis is typical of the genus Lamtostyla: parental structures are involved in the formation of frontoventral transverse anlagen in the proter. Phylogenetic analyses based on small subunit ribosomal DNA sequence data reveal that the systematic position of Lamtostyla is rather unstable with low support values across the tree. However, a well-recognized close relationship between Lamtostyla and Bistichella are shown in all phylogenetic analyses.

Morphology, morphogenesis and molecular phylogeny of a new marine ciliate, Trichototaxis marina n. sp. (Ciliophora, Urostylida).

The live morphology, infraciliature and morphogenesis of a new urostylid ciliate, Trichototaxis marina n. sp., collected from coastal water in Qingdao, China, were studied based on the observations of live and silver stained specimens. The new species is characterised as follows: body very flexible and contractile, slight to brick-reddish in colour due to irregularly-shaped, brick-red pigments; ca. 70 adoral membranelles; about 17 frontal cirri arranged in a bicorona; average 67 midventral pairs, the right base of each pair being conspicuously larger than the left base; five to seven transverse cirri; constantly two frontoterminal, one buccal and two pretransverse ventral cirri; two or three left marginal rows; right and innermost left marginal rows with 56-92 and 66-106 cirri, respectively; six bipolar dorsal kineties; more than 100 macronuclear nodules. The characteristic morphogenetic feature in T. marina is the development of the left marginal rows, that is, only one left marginal row is newly built the other one or two being retained from the parental cell. Phylogenetic analyses based on small subunit ribosomal gene sequence data reveal a close relationship of T. marina with members of family Pseudokeronopsidae.