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Ciliophora are one of the most diverse phyla of unicellular eukaryotes. Members of this group are widely distributed and have been found in several environments including those that are anaerobic. However, the paucity of sufficient omics data for the major classes of the phylum Ciliophora, especially the anaerobic ciliate lineages, has been a serious impediment to research thus far. In this study, omics data were expanded to include representatives of 16 ciliate classes (excluding Cariacotrichea and Copemetopea) using single‐cell sequencing techniques. Using omics data of 57 ciliates and two apicomplexans, we conducted a comparative genomics study. Phylogenomic analysis based on 156 orthologous genes showed that: there are two superclades within the subphylum Intramacronucleata, i.e., Colpodea + Oligohymenophorea + Nassophorea + Plagiopylea + Prostomatea + Phyllopharyngea (CONThreeP), and Spirotrichea + Armophorea + Litostomatea + Odontostomatea + Muranotrichea + Parablepharismea (SALOMP); with the exception of Armophorea, all ciliate classes are monophyletic; neither Metopus nor Plagiopyla is monophyletic; Paraspathidium has a close affiliation with prostomateans. The results of the divergence time estimations indicated that: the radiation of Ciliophora occurred ca. 1067 million years ago (Mya); the SALOMP and CONThreeP lineages diverged from the intramacronucleatean common ancestor ca. 892 Mya and 851 Mya, respectively; the split between Muranotrichea and Parablepharismea occurred ca. 712 Mya; Odontostomatea are the youngest class, with a crown age estimated at ca. 19 Mya. In addition, we profiled stop codon usage and programmed ribosomal frameshifting (PRF) events. Our analyses revealed that: most of the species investigated use either TAA or TGA as the biased stop codon; stop codon usage in Paraspathidium sp. resembles that of Prostomatea; almost all ciliates included in the analyses have prevalent +1 and −1 PRF events. Finally, through GO enrichment analysis for group‐shared orthogroups of five anaerobic ciliate classes, we successfully identified the biological process and molecular function GO terms that were related to the adaptation of this particular group of ciliates to anaerobic environments. Together, these findings provide new insights into the evolutionary history of the Ciliophora and deepen our understanding of the adaptation of anaerobic ciliate groups to anaerobic environments from an evolutionary perspective. [ABSTRACT FROM AUTHOR]

Most eukaryotes have one nucleus and nuclear genome per cell. Ciliates have instead evolved distinct nuclei that coexist in each cell: a silent germline vs. transcriptionally active somatic nuclei. In the best-studied model species, both nuclei can divide asexually, but only germline nuclei undergo meiosis and karyogamy during sex. Thereafter, thousands of DNA segments, called internally eliminated sequences (IESs), are excised from copies of the germline genomes to produce the streamlined somatic genome. In Loxodes, however, somatic nuclei cannot divide but instead develop from germline copies even during asexual cell division, which would incur a huge overhead cost if genome editing was required. Here, we purified and sequenced both genomes in Loxodes magnus to see whether their nondividing somatic nuclei are associated with differences in genome architecture. Unlike in other ciliates studied to date, we did not find canonical germline-limited IESs, implying Loxodes does not extensively edit its genomes. Instead, both genomes appear large and equivalent, replete with retrotransposons and repetitive sequences, unlike the compact, gene-rich somatic genomes of other ciliates. Two other hallmarks of nuclear development in ciliates--domesticated DDE-family transposases and editing-associated small RNAs--were also not found. Thus, among the ciliates, Loxodes genomes most resemble those of conventional eukaryotes. Nonetheless, base modifications, histone marks, and nucleosome positioning of vegetative Loxodes nuclei are consistent with functional differentiation between actively transcribed somatic vs. inactive germline nuclei. Given their phylogenetic position, it is likely that editing was present in the ancestral ciliate but secondarily lost in the Loxodes lineage. [ABSTRACT FROM AUTHOR]

Unicellular ciliates like Tetrahymena are best known as free-living bacteriovores, but many species are facultative or obligate parasites. These "histophages" feed on the tissues of hosts ranging from planarian flatworms to commercially important fish and the larvae of imperiled freshwater mussels. Here, we developed a novel bioinformatics pipeline incorporating the nonstandard ciliate genetic code and used it to search for Ciliophora sequences in 34 publicly available Platyhelminthes EST libraries. From 2,615,036 screened ESTs, we identified nearly 6,000 high-confidence ciliate transcripts, supporting parasitism of seven additional flatworm species. We also cultured and identified Tetrahymena from nine terrestrial and freshwater planarians, including invasive earthworm predators from the genus Bipalium and the widely studied regeneration models Dugesia japonica and Schmidtea mediterranea. A co-phylogenetic reconstruction provides strong evidence for the coevolution of histophagous Ciliophora with their Platyhelminthes hosts. We further report the antiprotozoal aminoglycoside paromomycin expels Tetrahymena from S. mediterranea , providing new opportunities to investigate the effects of this relationship on planarian biology. Together, our findings raise the possibility that invasive flatworms constitute a novel dispersal mechanism for Tetrahymena parasites and position the Platyhelminthes as an ideal model phylum for studying the ecology and evolution of histophagous ciliates. [ABSTRACT FROM AUTHOR]

[Display omitted] • Phylogenomics, divergence time estimation and macroevolutionary analyses are conducted on 105 ciliate species and 247 genes. • A robust reference tree for the phylum Ciliophora. • The radiation of Ciliophora occurred around 1175 Ma in the middle of the Proterozoic Eon. • Most classes diverged from their sister lineages during the latter half of the Proterozoic Eon. • The variation in species richness among classes is primarily attributed to net diversification rates rather than age. Ciliophora, an exceptionally diverse lineage of unicellular eukaryotes, exhibits a remarkable range of species richness across classes in the ciliate Tree of Life. In this study, we have acquired transcriptome and genome data from 40 representative species in seven ciliate classes. Utilizing 247 genes and 105 taxa, we devised a comprehensive phylogenomic tree for Ciliophora, encompassing over 60 % of orders and constituting the most extensive dataset of ciliate species to date. We established a robust phylogenetic framework that encompasses ambiguous taxa and the major classes within the phylum. Our findings support the monophyly of each of two subphyla (Postciliodesmatophora and Intramacronucleata), along with three subclades (Protocruzia, CONTHREEP, and SAPML) nested within Intramacronucleata, and elucidate evolutionary positions among the major classes within the phylum. Drawing on the robust ciliate Tree of Life and three constraints, we estimated the radiation of Ciliophora around 1175 Ma during the middle of the Proterozoic Eon, and most of the ciliate classes diverged from their sister lineage during the latter half of this period. Additionally, based on the time-calibrated tree and species richness pattern, we investigated net diversification rates of Ciliophora and its classes. The global net diversification rate for Ciliophora was estimated at 0.004979 species/Ma. Heterogeneity in net diversification rates was evident at the class level, with faster rates observed in Oligohymenophorea and Spirotrichea than other classes within the subclades CONTHREEP and SAPML, respectively. Notably, our analysis suggests that variations in net diversification rates, rather than clade ages, appear to contribute to the differences in species richness in Ciliophora at the class level. [ABSTRACT FROM AUTHOR]

Rustic aquaculture systems represent a sustainable production alternative in the world, providing appropriate conditions for the establishment of epibiotic ciliates on crayfish. However, the impact of epibiotic ciliates on the aquaculture production has been insufficiently studied hitherto. The objective of this work was to identify the epibiotic ciliates on the surface of the crayfish Procambarus (Austrocambarus) sp. and analyze whether there is any correlation between seasonality and environmental conditions in an artificial pond in Chiapas, Mexico. The crayfish were collected during dry and rainy seasons to investigate their ciliate epibionts. Some physical and chemical parameters of the sampled water were measured and correlation matrix analyses were applied to detect possible relationships between the presence of ciliates and environmental parameters. Thirteen species belonging to the subclasses Peritrichia and Suctoria were identified. Significant differences were found for some environmental parameters and species recorded in both seasons. The present data could help to better understand the relationship bewteen the epibiotic ciliate community on crayfish and some environmental parameters in aquaculture systems. [ABSTRACT FROM AUTHOR]

The spatial patterns of planktonic ciliate communities were studied from May to June 2019 in the Nile Delta's Damietta region, southeastern Mediterranean. The ciliate communities were sampled from twenty-five sites of five stressed domains with spatial gradients of environmental status. A total of 32 ciliate taxa with six dominant species were identified, comprising 21 tintinnids and 11 aloricate ciliates. The abundance and richness of each ciliate group varied geographically and were most strongly influenced by salinity variations; tintinnid ciliates attained high abundance and richness at high salinity sites in the harbour and coastal region and decreased within the estuary upstream. Aloricate ciliates were poorly represented at most sites but were a substantial proportion of upstream estuarine sites. Multivariate/univariate analyses demonstrated that spatial patterns of the ciliate communities were significantly correlated with environmental variables, especially salinity, chlorophyll-a, and nutrients, either alone or in combination with one another. These results indicate that the ciliates can be useful bioindicators in stressed environments while also allowing the detection of impacts on short time scales by rapidly responding to environmental variations., (© 2024. The Author(s).)

Ciliated protozoa (ciliates) are an ecologically important group of microeukaryotes that play roles in the flow of energy and nutrients in aquatic and terrestrial ecosystems. The community distribution and diversity of soil ciliates in the Nianchu River Basin were investigated by sampling four major habitats, i.e., grassland, farmland, wetland and sea buckthorn forest during May, August and October 2020. Cultivation identification and enumeration of soil ciliates were performed by the non-submerged culture method, in vivo observations and protargol silver staining, and direct counting methods, respectively. A total of 199 species were identified representing, 89 genera, 67 families, 31 orders and 11 classes. Haptorida was the dominant group with 35 species, accounting for 17.59% of the total. The results showed that the α and β diversity indices of soil ciliate communities in the Nianchu River Basin varied significantly in spatial distribution, but not in temporal distribution. Mantel test showed that soil water content, total nitrogen and organic matter were significantly correlated with soil ciliates. Soil water content was the main environmental factor driving the spatial distribution of soil ciliates. Co-occurrence network analysis showed that soil ciliate species in the Nianchu River Basin depend on each other in the relationship of solidarity and cooperation or ecological complementarity. Thus maintaining or enhancing the diversity and stability of the community. Community assembly shows that randomness process was an important ecological process driving soil ciliate community construction in the Nianchu River Basin., Competing Interests: The authors declare no competing interests., (Copyright: © 2024 Zhu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Ciliate protozoa are key members of the microbial community of the rumen. Their study is important to the health and productivity of cattle, which are their hosts. However, there have been persistent challenges in culturing this microbial group in the laboratory. This review will sum up recent advances along with these persistent challenges. Protozoa have been maintained in three types of cultures (ex vivo, in vitro batch, in vitro continuous). Ex vivo cultures are prepared readily from rumen contents by washing away contaminating cells (e.g., bacteria). They have been useful in making basic observations of metabolism, such as which types of fermentation products protozoa form. However, these cultures can be maintained for only short periods (minutes or hours). In vitro batch and in vitro continuous cultures can be used in longer experiments (weeks or longer). However, it is not currently possible to maintain protozoa in these cultures unless bacteria are also present. We conclude the review with a protocol for preparing ex vivo cultures of protozoa. Our protocol has been standardized and used successfully across animal diets, users, and institutions. We anticipate this review will prepare others to culture rumen ciliate protozoa and reach new insights into this important microbial group., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Exploring the mechanisms that underpin symbiosis requires an understanding of how these complex interactions are maintained in diverse model systems. The ciliate protist, Paramecium bursaria, offers a valuable insight into how emergent endosymbiotic interactions have evolved., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Benjamin H. Jenkins. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Protozoan ciliates represent a common biological contaminant during microalgae cultivation, which will lead to a decline in microalgae productivity. This study investigated the effectiveness of sodium dodecyl benzene sulfonate (SDBS) in controlling ciliate populations within microalgae cultures. SDBS concentrations of 160 mg/L and 100 mg/L were found to effectively manage the representative species of ciliates contamination by Euplotes vannus and Uronema marinum during the cultivation of Synechococcus and Chlorella, and the growth vitality of microalgae has been restored. Additionally, SDBS at these concentrations reduced oxidative stress resistance and induced membrane damage to remove biological pollutants by modulating enzyme activity, affecting lipid, energy, amino acid metabolism pathways, and processes such as translation and protein folding. This research provides insights into the mechanisms through which SDBS effectively combats protozoan ciliates during the microalgal cultivation. This contributes to reduce biological pollution, ensure the overall productivity and healthy and sustainable management of microalgae ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Since their discovery, rumen ciliates of domestic cattle have been reported from various geographic locations. However, until now there is only one taxonomic inventory of ciliates associated with Brazilian cattle. The present study aimed to assess the community composition, relative abundance, richness, and density of rumen ciliates in Brazilian cattle, whose feeding diets were supplemented with crescent urea levels. Across all treatments analyzed, one subclass, two orders, three families, 11 genera, and 31 species of ciliates were identified. The ciliate community composition and species richness varied among the four treatments used. However, the total ciliate density was not affected by the experimental diets. We described a new entodiniid morphotype, Entodinium caudatum m. orbonuclearis, and recorded Oligoisotricha bubali for the second time in Brazil., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

Takifugu rubripes (T. rubripes) is a valuable commercial fish, and Cryptocaryon irritans (C. irritans) has a significant impact on its aquaculture productivity. DNA methylation is one of the earliest discovered ways of gene epigenetic modification and also an important form of modification, as well as an essential type of alteration that regulates gene expression, including immune response. To further explore the anti-infection mechanism of T. rubripes in inhibiting this disease, we determined genome-wide DNA methylation profiles in the gill of T. rubripes using whole-genome bisulfite sequencing (WGBS) and combined with RNA sequence (RNA-seq). A total of 4659 differentially methylated genes (DMGs) in the gene body and 1546 DMGs in the promoter between the infection and control group were identified. And we identified 2501 differentially expressed genes (DEGs), including 1100 upregulated and 1401 downregulated genes. After enrichment analysis, we identified DMGs and DEGs of immune-related pathways including MAPK, Wnt, ErbB, and VEGF signaling pathways, as well as node genes prkcb, myca, tp53, and map2k2a. Based on the RNA-Seq results, we plotted a network graph to demonstrate the relationship between immune pathways and functional related genes, in addition to gene methylation and expression levels. At the same time, we predicted the CpG island and transcription factor of four immune-related key genes prkcb and mapped the gene structure. These unique discoveries could be helpful in the understanding of C. irritans pathogenesis, and the candidate genes screened may serve as optimum methylation-based biomarkers that can be utilized for the correct diagnosis and therapy T. rubripes in the development of the ability to resist C. irritans infection., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

In the 1980s, a mass die-off of the long-spined sea urchin Diadema antillarum occurred on Florida and Caribbean coral reefs. D. antillarum populations largely did not recover, and in 2022, remaining populations experienced another mass mortality event. A ciliate most similar to Philaster apodigitiformis was identified as the causative agent of the 2022 event, which was named D. antillarum scuticociliatosis (DaSc). Here, we investigated possible treatments for this pathogen. We tested the efficacy of 10 compounds at final concentrations of 100, 50, 25, 12.5, 6.25, and 3.13 µM, or a 10-fold serial dilution series, against ciliates cultured from an infected D. antillarum specimen. Of the tested compounds, 8 induced 100% ciliate mortality at some dose after 24 h. The most effective (defined as those requiring the lowest dose to induce 100% ciliate mortality) were quinacrine and tomatine (both effective at 12.5 µM), followed by furaltadone and plumbagin (25 µM), bithionol sulfoxide and 2'4' dihydroxychalcone (50 µM), and oxyclozanide and carnidazole (100 µM). Toltrazuril and a commercially available anticiliate product containing naphthoquinones were not effective at any dose tested. Shortened (15 min) time trials were performed using ciliate cultures reared in natural seawater to better reflect natural environmental conditions, and revealed that 2 of the compounds (quinacrine and tomatine) induced 100% ciliate mortality at 100 µM, with tomatine also effective at 50 µM. This study identified several treatments effective against the causative agent of DaSc in vitro, but their toxicity and utility in vivo remain unknown.

In tidal flats, which are located at the transition zone between terrestrial and marine ecosystems, environmental factors such as temperature, sediment particle size, and tidal range exhibit geographic variation. Accordingly, the composition and structure of the microbial communities in the tidal flats are likely to vary in geographically different habitats. To clarify these differences with environmental factors causing them, we analyzed microbial communities consisting of bacteria and ciliates in sediments collected from nine tidal flats in geographical diverse region from Hokkaido to Kagoshima, Japan. The results confirmed that the community structures of bacteria and ciliophora in tidal flat sediments differed at the geographical scale of the Japanese archipelago. However, the variation could not be explained by the physical distance between the tidal flats nor by the differences in the trophic conditions among the tidal flats. Instead, the OTU richness of both the bacterial and ciliophoran communities was significantly related to the tidal range. The results also showed that bacteria and ciliophora tended to form similar communities among the tidal flats with similar median particle sizes. Furthermore, ciliophoran communities were similar among the tidal flats with similar bacterial communities. The results suggest that bacteria and ciliophora interact each other through trophic relationships or physical and chemical processes in the sediment habitats.

The latitudinal dynamics of biodiversity has been the focus of global attention. This study is based on the latitude gradient of biodiversity in the spatial changes of pelagic ciliate communities in the western Arctic Ocean. The gradient pattern of pelagic ciliate communities across four latitudes were investigated from the water surface at 22 sampling station in the northern Bering Sea of the western Arctic Ocean and Chukchi Sea from August 5 to August 24, 2016. Based on multivariate analyses, the results showed that (1) the spatial patterns of pelagic ciliates represented a significant latitudinal gradient along the western Arctic Ocean; (2) the species number and abundance of pelagic ciliate communities declined from 64°N to 80°N; (3) variations in the horizontal distribution of ciliates were significantly correlated with changes in physicochemical variables, especially water temperature and Chl a; Thus it is suggested that the expected latitudinal decline of biodiversity was evident along the western Arctic Ocean., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

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 in vivo 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., (© 2023 International Society of Protistologists.)

The subclass Peritrichia includes sessile ciliates that attach to aquatic non-living or living substrates. In the case of the latter, they live in a relationship called epibiosis. The genus Epistylis Ehrenberg,1830 is the second-most speciose genus within Sessilida, being characterized by its colonial lifestyle and having a non-contractile stalk. Species of Epistylis have been described from a wide range of aquatic habitats worldwide. The main goal of the present study is to provide a global checklist of the valid species of Epistylis, which may serve as a first step towards a taxonomic review of the genus.

The oceanic-to-neritic species shift of microzooplanktonic tintinnids and their interaction with relevant abiotic variables are two crucial processes in the marine ecosystem. However, these processes remain poorly documented in China's marginal seas. In the summer of 2022, we investigated the community structure of pelagic tintinnids in surface waters from the South China Sea (SCS) to the Yellow Sea (YS), passing through the East China Sea (ECS). A number of 58 species from 23 genera were identified, with 36 and 22 species belonging to oceanic and neritic genera, respectively. The abundance proportion of oceanic and neritic genera exhibited a decreasing and increasing trend, respectively, from the SCS to YS. Furthermore, four distinctive tintinnid community groups were classified based on cluster analysis using tintinnid species and abundance data, and the position of southern Taiwan Strait was identified as the "Shift Point" for oceanic-to-neritic species dominance. The top two tintinnid species in each group showed distinct variations in body size. Additionally, multivariate biotic-abiotic statistical analyses revealed that temperature determined tintinnid species richness, while temperature, salinity, Si(OH) 4 , and Chl a determined tintinnid abundance. Our study provides a substantial foundation for recognizing the oceanic-to-neritic species shift of tintinnids in the China's marginal seas, and highlights the role of biotic-abiotic factors in driving biogeochemical fluxes and the potential response of microzooplankton to future climate change., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Cryptocaryoniasis remains a major parasitic disease and economic challenge for marine aquaculture. Cryptocaryoniasis in marine fish is caused by Cryptocaryon irritans (Ciliata). A theront is a motile, free-swimming stage in the life cycle of C. irritans, which is typically the infective stage that actively seeks out a host to initiate infection. Population density and growth rate of theronts were investigated in Nile tilapia, Oreochromis niloticus fed with citric acid-supplemented feed. The experiment involved feeding three diets with graded levels of citric acid (0, control diet, 0.5, 1 and 1.5 g kg -1 diet), to seawater-adapted Nile tilapia (O. niloticus) juveniles for 21 days. The results showed that citric acid in the fish feed had an impact on the theront number of C. irritans in a manner of dose-dependent. In the experimental cohort administered a diet supplemented with 1.5 g kg -1 citric acid, the population density of theronts was observed to be significantly reduced, measured at 29 ± 3.34, as opposed to 473.34 ± 16.48 in the control group at the culmination of the experiment. The observed population growth rate of theronts was significantly higher in the control group than in the group administered the citric acid feed (p < .005). The growth rate (r d -1 ) was 0.12 in control, 0.05 in 0.5 g kg -1 , 0.031 in 1 g kg -1 , and - 0.031 in 1.5 g kg -1 citric acid-supplemented groups. Fish growth and feed conversion ratio were not affected by the citric acid in the feed. In conclusion, the findings of this investigation provide a valuable addition to our understanding of the potential protective effects of citric acid supplementation for fish against the C. irritans parasite. This is evidenced by the observed reduction in theronts present in the water., (© 2023 John Wiley & Sons Ltd.)

We examined the effects of light exposure on the theront excystment circadian rhythm in Cryptocaryon irritans using a newly invented apparatus, which enabled us to examine the excystment rhythms of theronts from tomonts with low labor. Using the apparatus, we examined the timings of theront excystment from tomonts exposed directly to light and from tomonts exposed to light-exposed seawater by counting the number of excysted theronts. We found that the theront excystment time changed only when tomonts were directly exposed to light, indicating that light reception is essential for circadian rhythm entrainment. When tomonts were exposed to light only once for 12 h, either on Day 1, Day 2, or Day 3 after leaving host and being encysted, the circadian rhythm was entrained according to the photoperiod given on tomonts. Tomonts exposed to a low light (1 lx) with 12L:12D photoperiod daily showed a circadian rhythm similar to that in tomonts exposed to an intense light (500 lx) under the same photoperiod. When tomonts were incubated at 22 °C, 25 °C, or 28 °C under the same photoperiod, almost the same circadian rhythm was developed, suggesting temperature has little effect on the circadian rhythm entrainment between the range, even though the date of excystment was delayed in lower temperatures. These results suggest the circadian rhythm of theront excystment can be entrained in tomonts on the seabed of inner bays where net-cage aquaculture is conducted, and be involved in the outbreaks of cryptocaryoniasis there., Competing Interests: Declaration of Competing Interest Declaration of generative AI and AI-assisted technologies in the writing process. During the preparation of this work the authors used Chat GPT in order to edit the manuscript. After using this tool, the authors reviewed and edited the content as needed and take full responsibility for the content of the publication., (Copyright © 2023. Published by Elsevier B.V.)

Soil and plant-associated protistan communities play a key role in shaping bacterial and fungal communities, primarily through their function as top-down predators. However, our understanding of how pathogen invasion influences these protistan communities and their relationships with bacterial and fungal communities remains limited. Here, we studied the protistan communities along the soil-plant continuum of healthy chilli peppers and those affected by Fusarium wilt disease (FWD), and integrated bacterial and fungal community data from our previous research. Our research showed that FWD was associated with a significant enrichment of phagotrophic protists in roots, and also increased the proportion and connectivity of these protists (especially Cercozoa and Ciliophora) in both intra- and inter-kingdom networks. Furthermore, the microbiome of diseased plants not only showed a higher relative abundance of functional genes related to bacterial anti-predator responses than healthy plants, but also contained a greater abundance of metagenome-assembled genomes with functional traits involved in this response. The increased microbial inter-kingdom associations between bacteria and protists, coupled with the notable bacterial anti-predator feedback in the microbiome of diseased plants, suggest that FWD may catalyse the associations between protists and their microbial prey. These findings highlight the potential role of predatory protists in influencing microbial assembly and functionality through top-down forces under pathogenic stress., (© 2023 John Wiley & Sons Ltd.)

Protozoan fauna is playing an important role in the functioning of microbial food webs by transferring the flux of material and energy from low to high tropic levels in marine ecosystems. To assess effects of elevated temperature on the marine ecosystem, periphytic protozoan communities were used as the test microbial fauna, and were incubated in a temperature-controlled circulation system in a successive temperature gradient of 22 (control), 25, 28, 31 and 34 °C. The results showed that: (1) the test microbial fauna was shifted in both species composition and community structure; (2) the average taxonomic distinctness represented a clear decreasing trend, (3) while the variation in taxonomic distinctness significantly increased with increase of water temperature; and (4) the community pattern was significantly departed from an expectation when temperature increased by 12 °C. These results suggested that Protozoa may be used as a useful bioindicator of global warming in marine ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: As unicellular eukaryotes, ciliates are an indispensable component of micro-ecosystems that play the role of intermediate nutrition link between bacteria or algae and meiofauna. Recent faunistic studies have revealed many new taxa of hypotrich ciliates, indicating their diversity is greater than previously thought. Here we document an undescribed form isolated from an artificial brackish water pond in East China. Examination of its morphology, ontogenesis and molecular phylogeny suggests that it represents a new species., Results: The morphology and morphogenesis of the new brackish-water deviatid ciliate, Heterodeviata nantongensis nov. sp., isolated from Nantong, China, were investigated using live observations and protargol staining. The diagnostic traits of the new species include three frontal cirri, one buccal cirrus, one or two parabuccal cirri, an inconspicuous frontoventral cirral row of four to six frontoventral cirri derived from two anlagen, three left and two right marginal rows, two dorsal kineties, dorsal kinety 1 with 9-14 dikinetids and dorsal kinety 2 with only two dikinetids, and one to three caudal cirri at the rear end of dorsal kinety 1. Its main morphogenetic features are: (i) the old oral apparatus is completely inherited by the proter except undulating membranes, which are reorganized in situ; (ii) anlagen for marginal rows and the left dorsal kinety develop intrakinetally in both proter and opisthe; (iii) dorsal kinety 2 is generated dorsomarginally; (iv) five cirral anlagen are formed in both proter and opisthe; (v) in the proter, anlagen I and II very likely originate from the parental undulating membranes and the buccal cirrus, respectively, anlage III from anterior parabuccal cirrus, anlage IV originates from the parental frontoventral cirri and anlage V from the innermost parental right marginal row; and (vi) anlagen I-IV of the opisthe are all generated from oral primordium, anlage V from the innermost parental right marginal row. Phylogenetic analyses based on SSU rRNA gene sequence data were performed to determine the systematic position of the new taxon., Conclusions: The study on the morphology, and ontogenesis of a new brackish-water taxon increases the overall knowledge about the biodiversity of this ciliate group. It also adds to the genetic data available and further provides a reliable reference for environmental monitoring and resource investigations., (© 2024. The Author(s).)

The freshwater raphidophyte Gonyostomum semen forms extensive summer blooms in northern European humic lakes. The development of these blooms might be facilitated by a lack of natural top-down control, as few zooplankton species are able to prey on these large algal cells (up to 100 μm) that expel trichocysts upon physical stress. In this study, we describe a small ciliate species (< 17 μm) that preys on G. semen by damaging the cell membrane until cytoplasm and organelles spill out. Sequencing of clonal cultures of the ciliate tentatively identified it as the prostomatid species Urotricha pseudofurcata. Grazing experiments illustrated that feeding by U. cf. pseudofurcata can significantly reduce cell concentrations of the microalga. However, differences in cell size and growth rate between two investigated ciliate strains resulted in noticeably different grazing pressure. Environmental sequencing data from five different lakes supported potential interactions between the two species. Urotricha cf. pseudofurcata might, thus, play an important role in aquatic ecosystems that are regularly dominated by G. semen, reducing the abundance of this bloom-forming microalga and enabling transfer of organic carbon to higher trophic levels., (© 2024. The Author(s).)

Changes in the biodiversity of aquatic environments over time and space due to human activities are a topic of theoretical and conservational interest in ecology. Thus, variation in taxonomic beta diversity of the planktonic ciliates community was investigated along a temporal and spatial gradient in two subsystems of a Neotropical floodplain, one impacted by dams (Paraná) and the other free of them along its course (Ivinhema). For the spatial analysis, the Paraná subsystem did not show a significant decrease in beta diversity, presenting a pattern like that observed for the Ivinhema subsystem. Therefore, biotic homogenization was not observed for the ciliate's community downstream of the dams. It was noted that there was a fluctuation in the relevance of the components of beta diversity, regardless of the subsystem analyzed. For the temporal analysis there was a significant change in species composition from the first to the last year investigated, essentially for the subsystem impacted by dams, and that this was determined mainly by species loss. Although spatial beta diversity remained high without a clear process of biotic homogenization, dams promoted remarkable changes in ciliate species composition over the years mainly by continuous loss of species., (© The Author(s) 2024. Published by Oxford University Press on behalf of FEMS.)

Free-living litostomatean ciliates, prominent microeukaryote predators commonly encountered in freshwater and marine habitats, play vital roles in maintaining energy flow and nutrient cycles. Nevertheless, understanding their biodiversity and phylogenetic relationships remains challenging due to insufficient morphological information and molecular data. As a new contribution to this group, three haptorian ciliates, including two new species (Actinobolina bivacuolata sp. nov. and Papillorhabdos foissneri sp. nov.) and the insufficiently described type species, Actinobolina radians, were isolated from wetlands around Lake Weishan, China and investigated by a combination of living morphology, stained preparations, and 18S rRNA gene sequence data. An illustrated key of the valid species within the two genera is provided. In addition, we reveal the phylogenetic positions of these two genera for the first time. Although they differ in all key morphologic characters such as general appearance (ellipsoidal with numerous tentacles vs. cylindrical), extrusomes (stored in tentacles vs. anchored to pellicle), circumoral kinety (present vs. absent), composition of somatic kineties (kinetosome clusters vs. monokinetids), and number of dorsal brush rows (1 vs. 4), they both cluster in a fully supported clade in the phylogenetic tree, which indicates that the biodiversity and additional molecular markers of this group need further exploration., (© 2023 International Society of Protistologists.)

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., (© 2023 The Authors. Journal of Eukaryotic Microbiology published by Wiley Periodicals LLC on behalf of International Society of Protistologists.)

To investigate effects of salinity variability on colonization dynamics of periphytic protozoan fauna, a 21-day study was conducted in temperature-controlled circulation systems (TCCSs). Periphytic protozoan communities were incubated using glass slides as artificial substrata in five TCCS aquaria with a large-scale salinity gradient of 9, 19, 29 (control), 39, and 49 PSU, respectively. The colonization dynamics were observed on days 3, 5, 7, 10, 14, and 21. The colonization dynamics were well fitted to the MacArthur-Wilson and logistic model equations in colonization and growth curves in all five treatments, respectively. However, the maximum species richness and abundance were reduced, and the colonization patterns were significantly shifted in four treatments with salinity changed by 20 PSU compared to the control (29 PSU). Thus, it is suggested that the large-scale salinity variability may reduce the species richness significantly and affect colonization dynamics of periphytic protozoan fauna in marine environments., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Microplastics (MPs), due to their micro size, which overlaps with the typical food size of various aquatic organisms, can be ingested and move up the food chain, accumulating in the bodies of organisms at higher trophic levels. Few studies have focused on the uptake of MPs by ciliates, which are an important element of the microbial cycle. Three different ciliate species were used in this study: Blepharisma japonicum, Euplotes sp., and Spirostomum teres, as well as polystyrene beads with diameters of 1 and 2 µm at two concentrations (10 6 and 10 7 beads × mL -1 ). The results of the experiments showed that MPs have a variable, species-specific effect on the population growth rate of ciliates, which is directly dependent on their concentration in the environment (P < 0.01). It was also observed that the number of MPs ingested changed over time depending on their concentration and size. On average, the highest number of ingested MPs (883.11 ± 521.47) was recorded at 60 min of exposure to a low concentration of small beads in B. japonicum. The lowest number of beads was ingested after 5 min of exposure to a low concentration of large beads in the same species. The rate of MP uptake by the ciliate species was significantly dependent on their concentration, exposure time, and size (P < 0.001). The highest clearance rate was observed in the fifth minute of the experiment in the environment with the lowest MP concentration., (© 2023. The Author(s).)

Paramecium (Ciliophora, Oligohymenophorea) is a good model to study ciliate biogeography. Extensive sampling mainly in northern hemisphere has led to 16 valid morphological species description thus far. However, a majority of hard-to-reach regions, including South East Asia, are underinvestigated. Our study combined traditional morphological and molecular approaches to reveal the biodiversity of Paramecium in Thailand from more than 110 samples collected in 10 provinces. Representatives of seven morphological species were identified from our collection, including the rare species, such as P. gigas and P. jenningsi. Additionally, we detected five different sibling species of the P. aurelia complex, described a new cryptic species P. hiwatashii n. sp. phylogenetically related to P. caudatum, and discovered a potentially new genetic species of the P. bursaria species complex. We also documented a variety of bacterial cytoplasmic symbionts from at least nine monoclonal cultures of Paramecium., (© 2023 International Society of Protistologists.)