+Advanced Search
Volume 4 Issue 4
Nov.  2022
Article Contents

Citation:

Biodiversity of freshwater ciliates (Protista, Ciliophora) in the Lake Weishan Wetland, China: the state of the art

  • Corresponding author: Borong Lu, boronglu@126.com
  • Received Date: 2022-04-15
    Accepted Date: 2022-10-20
    Published online: 2022-11-28
  • Edited by Jiamei Li.
  • Special topic: Ciliatology.
  • Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
  • Ciliates are core components of the structure of and function of aquatic microbial food webs. They play an essential role in the energy flow and material circulation within aquatic ecosystems. However, studies on the taxonomy and biodiversity of freshwater ciliates, especially those in wetlands in China are limited. To address this issue, a project to investigate the freshwater ciliates of the Lake Weishan Wetland, Shandong Province, commenced in 2019. Here, we summarize our findings to date on the diversity of ciliates. A total of 187 ciliate species have been found, 94 of which are identified to species-level, 87 to genus-level, and six to family-level. These species show a high morphological diversity and represent five classes, i.e., Heterotrichea, Litostomatea, Prostomatea, Oligohymenophorea, and Spirotrichea. The largest number of species documented are oligohymenophoreans. A comprehensive database of these ciliates, including morphological data, gene sequences, microscope slide specimens and a DNA bank, has been established. In the present study, we provide an annotated checklist of retrieved ciliates as well as information on the sequences of published species. Most of these species are recorded in China for the first time and more than 20% are tentatively identified as new to science. Additionally, an investigation of environmental DNA revealed that the ciliate species diversity in Lake Weishan Wetland is higher than previously supposed.
  • 加载中
  • Abraham JS, Somasundaram S, Maurya S, Gupta R, Toteja R (2021) Characterization of Euplotes lynni nov. spec., E. indica nov. spec. and description of E. aediculatus and E. woodruffi (Ciliophora, Euplotidae) using an integrative approach. Eur J Protistol 79: 125779 doi: 10.1016/j.ejop.2021.125779
    Berger H (1999) Monograph of the Oxytrichidae (Ciliophora, Hypotrichia). Monogr Biol 78: 1–1080
    Berger H (2006) Monograph of the Urostyloidea (Ciliophora, Hypotricha). Monogr Biol 85: 1–1304
    Berger H (2011) Monograph of the Gonostomatidae and Kahliellidae (Ciliophora, Hypotricha). Monogr Biol 90: 1–740
    Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30: 2114–2120 doi: 10.1093/bioinformatics/btu170
    Bower SM, Carnegie RB, Goh B, Jones SR, Lowe GJ, Mak MW (2004) Preferential PCR amplification of parasitic protistan small subunit rDNA from metazoan tissues. J Eukaryot Microbiol 51: 325–332 doi: 10.1111/j.1550-7408.2004.tb00574.x
    Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Peña AG, Goodrich JK, Gordon JI (2010) QⅡME allows analysis of high-throughput community sequencing data. Nat Methods 7: 335–336 doi: 10.1038/nmeth.f.303
    Chen LL, Ren YJ, Han K, Stoeck T, Jiang JM, Pan HB (2022) Re-description and molecular phylogeny of the free-swimming peritrichs Hastatella radians Erlanger, 1890 and H. aesculacantha Jarocki & Jakubowska, 1927 (Ciliophora, Peritrichia) from China. Eur J Protistol 84: 125891 doi: 10.1016/j.ejop.2022.125891
    Chi Y, Li YQ, Zhang QQ, Ma MZ, Warren A, Chen XR, Song WB (2020) New contributions to two ciliate genera (Ciliophora, Heterotrichea) based on morphological and molecular analyses, with description of a new Gruberia species. BMC Microbiol 20: 297 doi: 10.1186/s12866-020-01879-4
    Chi Y, Chen XR, Li YQ, Wang CD, Zhang TT, Ayoub A, Warren A, Song WB, Wang YY (2021a) New contributions to the phylogeny of the ciliate class Heterotrichea (Protista, Ciliophora): analyses at family-genus level and new evolutionary hypotheses. Sci China Life Sci 64: 606–620 doi: 10.1007/s11427-020-1817-5
    Chi Y, Wang Z, Lu BR, Ma HG, Mu CJ, Warren A, Zhao Y (2021b) Taxonomy and phylogeny of the dileptid ciliate genus Paradileptus (Protista: Ciliophora), with a brief review and redescriptions of two species isolated from a wetland in northern China. Front Microbiol 12: 709566 doi: 10.3389/fmicb.2021.709566
    Chi Y, Wang Z, Ye TT, Wang Y, Zhao JL, Song WB, Bourland WA, Chen XR (2022a) A new contribution to the taxonomy and phylogeny of the ciliate genus Spirostomum (Alveolata, Ciliophora, Heterotrichea), with comprehensive descriptions of two species from wetlands in China. Water Biol Secur 1: 100031 doi: 10.1016/j.watbs.2022.100031
    Chi Y, Liu YJ, Ma HG, Wang Y, Liu R, Al-Farraj SA, Song WB, Bourland WA (2022b) Taxonomy and molecular phylogeny of two poorly known ciliate genera, Balantidion and Acropisthium (Protista: Ciliophora: Litostomatea), including a new species of Balantidion. Eur J Protistol 85: 125906 doi: 10.1016/j.ejop.2022.125906
    Chiang SC, Shen YF, Gong XJ (1983) Aquatic invertebrates of the Tibetan plateau. Science Press, Beijing
    Corliss JO (1953) Silver impregnation of ciliated protozoa by the Chatton-Lwoff technic. Stain Technol 28: 97–100 doi: 10.3109/10520295309105108
    Curds CR (1975) A guide to the species of the genus Euplotes (Hypotrichida, Ciliatea). Bull Br Mus Nat Hist (zool) 28: 1–61
    Edgar RC (2013) UPARSE: highly accurate OTU sequences from microbial amplicon reads. Nat Methods 10: 996–998 doi: 10.1038/nmeth.2604
    Edgar RC, Haas BJ, Clemente JC, Quince C, Knight R (2011) UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 27: 2194–2200 doi: 10.1093/bioinformatics/btr381
    Esteban GF, Fenchel TM (2021) Ecology of protozoa: the biology of free-living phagotrophic protists. Springer Nature, Berlin
    Finlay BJ (2002) Global dispersal of free-living microbial eukaryote species. Science 296: 1061–1063 doi: 10.1126/science.1070710
    Finlay BJ, Fenchel T (2004) Cosmopolitan metapopulations of free-living microbial eukaryotes. Protist 155: 237–244 doi: 10.1078/143446104774199619
    Foissner W (2012) Urotricha spetai nov. spec., a new plankton ciliate (Ciliophora, Prostomatea) from a fishpond in the Seidlwinkel Valley, Rauris, Austrian Central Alps. Verhandlungen Zool Bot Gesellschaft Österreich 148/149: 173–184
    Foissner W (2021) A detailed description of a Brazilian Holophrya teres (Ehrenberg, 1834) and nomenclatural revision of the genus Holophrya (Ciliophora, Prostomatida). Eur J Protistol 80: 125662 doi: 10.1016/j.ejop.2019.125662
    Foissner W, Berger H (1996) A user-friendly guide to the ciliates (Protozoa, Ciliophora) commonly used by hydrobiologists as bioindicators in rivers, lakes, and waste waters, with notes on their ecology. Freshw Biol 35: 375–482 doi: 10.1111/j.1365-2427.1996.tb01775.x
    Foissner W, Berger H (2021) Terrestrial ciliates (Protista, Ciliophora) from Australia and some other parts of the world. Ser Monogr Ciliophorae 5: i–xii, 1–380
    Foissner W, Oertel A (2009) Morphology and ciliary pattern of some rare haptorid ciliates, with a description of the new family Kamburophryidae (Protists, Haptoria). Eur J Protistol 45: 205–218 doi: 10.1016/j.ejop.2008.11.002
    Foissner W, Pfister G (1997) Taxonomic and ecologic revision of urotrichs (Ciliophora, Prostomatida) with three or more caudal cilia, including a user-friendly key. Limnologica 27: 311–348
    Foissner W, Blatterer H, Berger H, Kohmann F (1991) Taxonomische und ökologische Revision der Ciliaten des Saprobiensystems—Band Ⅰ: Cyrtophorida, Oligotrichida, Hypotrichia, Colpodea. Inf Bayer Landesamtes Wasserwirtschaft 1: 1–478
    Foissner W, Berger H, Kohmann F (1992) Taxonomische und okologische Revision der Ciliaten des Saprobiensystems. Band Ⅱ: Peritrichia, Heterotrichida, Odontostomatida. Inf Bayer Landesamtes Wasserwirtschaft 592: 1–502
    Foissner W, Berger H, Blatterer H, Kohmann F (1995) Taxonomische und Ökologische Revision der Ciliaten des Saprobiensystem, Band: Ⅳ Gymnostomatea, Loxodes, Suctoria. Inf Bayer Landesamtes Für Wasserwirtschaft 1: 1–540
    Foissner W, Berger H, Schaumburg J (1999) Identification and ecology of limnetic plankton ciliates. Inf Bayer Landesamtes Wasserwirtschaft 3: 1–793
    Foissner W, Chao A, Katz LA (2008a) Diversity and geographic distribution of ciliates (Protista: Ciliophora). Biodivers Conserv 17: 345–363 doi: 10.1007/s10531-007-9254-7
    Foissner W, Kusuoka Y, Shimano S (2008b) Morphology and gene sequence of Levicoleps biwae n. gen., n. sp. (Ciliophora, Prostomatida), a proposed endemic from the ancient Lake Biwa, Japan. J Eukaryot Microbiol 55: 185–200 doi: 10.1111/j.1550-7408.2008.00323.x
    Frantal D, Agatha S, Beisser D, Boenigk J, Darienko T, Dirren-Pitsch G, Filker S, Gruber M, Kammerlander B, Nachbaur L (2022) Molecular data reveal a cryptic diversity in the genus Urotricha (Alveolata, Ciliophora, Prostomatida), a key player in freshwater lakes, with remarks on morphology, food preferences, and distribution. Front Microbiol 12: 787290 doi: 10.3389/fmicb.2021.787290
    Gao F, Warren A, Zhang QQ, Gong J, Miao M, Sun P, Xu DP, Huang J, Yi ZZ, Song WB (2016) The all-data-based evolutionary hypothesis of ciliated protists with a revised classification of the phylum Ciliophora (Eukaryota, Alveolata). Sci Rep 6: 24874 doi: 10.1038/srep24874
    Gentekaki E, Kolisko M, Gong Y, Lynn D (2017) Phylogenomics solves a long-standing evolutionary puzzle in the ciliate world: the subclass Peritrichia is monophyletic. Mol Phylogenet Evol 106: 1–5 doi: 10.1016/j.ympev.2016.09.016
    Gómez F, Wang L, Lin S (2018) Morphology and molecular phylogeny of peritrich ciliate epibionts on pelagic diatoms: Vorticella oceanica and Pseudovorticella coscinodisci sp. nov. (Ciliophora, Peritrichia). Protist 169: 268–279 doi: 10.1016/j.protis.2018.03.003
    Gong YC, Yu YH, Villalobo E, Zhu FY, Miao W (2006) Reevaluation of the phylogenetic relationship between mobilid and sessilid peritrichs (Ciliophora, Oligohymenophorea) based on small subunit rRNA genes sequences. J Eukaryot Microbiol 53: 397–403 doi: 10.1111/j.1550-7408.2006.00121.x
    Guillou L, Bachar D, Audic S, Bass D, Berney C, Bittner L, Boutte C, Burgaud G, de Vargas C, Decelle J (2012) The protist ribosomal reference database (PR2): a catalog of unicellular eukaryote small sub-unit rRNA sequences with curated taxonomy. Nucleic Acids Res 41: D597–D604 doi: 10.1093/nar/gks1160
    Hines HN, McCarthy PJ, Esteban GF (2020) First records of 'flagship' soil ciliates in North America. Protist 171: 125739 doi: 10.1016/j.protis.2020.125739
    Hu XZ, Lin XF, Song WB (2019) Ciliate atlas: species found in the South China Sea. Science Press, Beijing
    Huang JB, Zhang TT, Zhang QQ, Li Y, Warren A, Pan HB, Yan Y (2018) Further insights into the highly derived haptorids (Ciliophora, Litostomatea): phylogeny based on multigene data. Zool Scr 47: 231–242 doi: 10.1111/zsc.12269
    Huang HC, Yang JP, Huang SX, Gu BW, Wang Y, Wang L, Jiao NZ, Xu DP (2021) Spatial distribution of planktonic ciliates in the western Pacific Ocean: along the transect from Shenzhen (China) to Pohnpei (Micronesia). Mar Life Sci Technol 3: 103–115 doi: 10.1007/s42995-020-00075-7
    Institute of Hydrobiology Chinese Academy of Sciences (1973) The fauna of pathogens in fishs in Hubei Province. Science Press, Beijing
    Jiang CQ, Wang GY, Xiong J, Yang WT, Sun ZY, Feng JM, Warren A, Miao W (2019) Insights into the origin and evolution of Peritrichia (Oligohymenophorea, Ciliophora) based on analyses of morphology and phylogenomics. Mol Phylogenet Evol 132: 25–35 doi: 10.1016/j.ympev.2018.11.018
    Jiang LM, Wang CC, Zhuang WB, Li S, Hu XZ (2021) Taxonomy, phylogeny, and geographical distribution of the little-known Helicoprorodon multinucleatum Dragesco, 1960 (Ciliophora, Haptorida) and key to species within the genus. Eur J Protistol 78: 125769 doi: 10.1016/j.ejop.2021.125769
    Jiang LM, Wang CC, Warren A, Ma HG, Hu XZ (2022) New considerations of the systematics of the family Holophryidae (Protozoa, Ciliophora, Prostomatea) with a description of Holophrya paradiscolor sp. nov. and a redescription of Pelagothrix plancticola. Syst Biodivers 20: 1–15
    Jung JH (2021) Taxonomy of four scuticociliates (Protozoa: Ciliophora) from coastal waters of South Korea. J Species Res 10: 184–190
    Jung JH, Omar A, Park MH, Nguyen TV, Jung YH, Yang HM, Min GS (2021) Anteholosticha foissneri n. sp., a marine hypotrich ciliate (Ciliophora: Spirotrichea) from Vietnam: morphology, morphogenesis, and molecular phylogeny. Eur J Protistol 78: 125768 doi: 10.1016/j.ejop.2021.125768
    Kahl A (1930) Urtiere oder Protozoa. I. Wimpertiere oder Ciliate (Infusoria). 1. Allgemeiner Teil und Prostomata. Die Tierwelt Der Angrenzenden Meeresteile 18: 1–180
    Kahl A (1931) Urtiere oder Protozoa. I. Wimpertiere oder Ciliata (Infusoria). 2. Holotricha. Die Tierwelt Und Der Angrenzenden Meeresteile 21: 181–398
    Kahl A (1932) Urtiere oder Protozoa I: Wimpertiere oder Ciliata (Infusoria) 3. Spirotricha Die Tierwelt Deutschlands 25: 399–650
    Kahl A (1933) Ciliata libera et ectocommensalia. In: Grimpe GEW (ed) Die Tierwelt der Nord-und Ostsee 23. Akademische Verlagsgesellschaft Becker & Erler, Leipzig
    Kahl A (1935) Urtiere oder Protozoa. I. Wimpertiere oder Ciliata (Infusoria). 4. Peritricha und Chonotricha. Die Tierwelt Der Angrenzenden Meeresteile 30: 651–886
    Klein BM (1958) The "dry" silver method and its proper use. J Protozool 5: 99–103 doi: 10.1111/j.1550-7408.1958.tb02535.x
    Kwon CB, Vd'ačný P, Shazib SUA, Shin MK (2014) Morphology and molecular phylogeny of a new haptorian ciliate, Chaenea mirabilis sp. n., with implications for the evolution of the dorsal brush in haptorians (Ciliophora, Litostomatea). J Eukaryot Microbiol 61: 278–292 doi: 10.1111/jeu.12105
    Li T, Pan XM, Lu BR, Miao M, Liu MJ (2021) Taxonomy and molecular phylogeny of a new freshwater ciliate Frontonia apoacuminata sp. nov. (Protista, Ciliophora, Oligohymenophorea) from Qingdao, PR China. Int J Syst Evol Microbiol 71: 005071
    Li T, Liu MJ, Warren A, Al-Farraj SA, Yi ZZ, Sheng YL (2022) Morphology and SSU rRNA gene-based phylogeny of three peniculid ciliates (Ciliophora, Oligohymenophorea) from China, including a new Frontonia species. Eur J Protistol 85: 125910 doi: 10.1016/j.ejop.2022.125910
    Liu WW, Shin MK, Yi ZZ, Tan YH (2021) Progress in studies on the diversity and distribution of planktonic ciliates (Protista, Ciliophora) in the South China Sea. Mar Life Sci Technol 3: 28–43 doi: 10.1007/s42995-020-00070-y
    Liu MJ, Liu YJ, Zhang TT, Lu BR, Gao F, Gu J, Al-Farraj SA, Hu XZ, Song WB (2022) Integrative studies on the taxonomy and molecular phylogeny of four new Pleuronema species (Protozoa, Ciliophora, Scuticociliatia). Mar Life Sci Technol 4: 179–200 doi: 10.1007/s42995-022-00130-5
    Lynn DH (2008) The ciliated protozoa: characterization, classification, and guide to the literature, 3rd edn. Springer, Dordrecht
    Ma HW, Choi JK, Song WB (2003) An improved silver carbonate impregnation for marine ciliated protozoa. Acta Protozool 42: 161–164
    Magoč T, Salzberg SL (2011) FLASH: fast length adjustment of short reads to improve genome assemblies. Bioinformatics 27: 2957–2963 doi: 10.1093/bioinformatics/btr507
    Mayén-Estrada R, Dias RJP (2021) A checklist of species of the family Zoothamniidae (Ciliophora: Peritrichia), symbionts of crustaceans. Zootaxa 4949: zootaxa. 4949.3.7
    McMurdie PJ, Holmes S (2013) phyloseq: an R package for reproducible interactive analysis and graphics of microbiome census data. PLoS ONE 8: e61217 doi: 10.1371/journal.pone.0061217
    Oksanen J, Blanchet FG, Kindt R, Legendre P (2014) Vegan: community ecology package. R package version 2.0-10
    Omar A, Ji HM, Jung JH (2021) Molecular phylogeny of a new gonostomatid ciliate revealing a discrepancy between interphasic and cell divisional patterns (Ciliophora, Hypotricha). Eur J Protistol 79: 125794 doi: 10.1016/j.ejop.2021.125794
    Paiva TDS (2020) Systematic redefinition of the Hypotricha (Alveolata, Ciliophora) based on combined analyses of morphological and molecular characters. Protist 171: 125755 doi: 10.1016/j.protis.2020.125755
    Pan HB, Zhang QQ, Dong JY, Jiang JM (2020) Morphology and phylogeny of two novel pleurostomatids (Ciliophora, Litostomatea), establishing a new genus. J Eukaryot Microbiol 67: 252–262 doi: 10.1111/jeu.12779
    Poláková K, Čepička I, Bourland WA (2021) Phylogenetic position of three well-known ciliates from the controversial order Loxocephalida Jankowski, 1980 (Scuticociliatia, Oligohymenophorea) and Urozona buetschlii (Schewiakoff, 1889) with improved morphological descriptions. Protist 172: 125833 doi: 10.1016/j.protis.2021.125833
    Pröschold T, Rieser D, Darienko T, Nachbaur L, Kammerlander B, Qian K, Pitsch G, Bruni EP, Qu ZS, Forster D, Rad-Menendez C, Posch T, Stoeck T, Sonntag B (2021) An integrative approach sheds new light onto the systematics and ecology of the widespread ciliate genus Coleps (Ciliophora, Prostomatea). Sci Rep 11: 1–19 doi: 10.1038/s41598-020-79139-8
    Safi LSL, Tang KW, Carnegie RB (2022) Investigating the epibiotic peritrich Zoothamnium intermedium Precht, 1935: seasonality and distribution of its relationships with copepods in Chesapeake Bay (USA). Eur J Protistol 84: 125880 doi: 10.1016/j.ejop.2022.125880
    Serra V, D'Alessandro A, Nitla V, Gammuto L, Modeo L, Petroni G, Fokin SI (2021a) The neotypification of Frontonia vernalis (Ehrenberg, 1833) Ehrenberg, 1838 and the description of Frontonia paravernalis sp. nov. trigger a critical revision of frontoniid systematics. BMC Zool 6: 4 doi: 10.1186/s40850-021-00067-9
    Serra V, Fokin SI, Gammuto L, Nitla V, Castelli M, Basuri CK, Satyaveni A, Sandeep BV, Modeo L, Petroni G (2021b) Phylogeny of Neobursaridium reshapes the systematics of Paramecium (Oligohymenophorea, Ciliophora). Zool Scr 50: 241–268 doi: 10.1111/zsc.12464
    Shao C, Chen XM, Jiang JM (2020) Hypotrichous ciliates in China. Science Press, Beijing
    Shazib SUA, Vd'ačný P, Kim JH, Jang SW, Shin MK (2014) Phylogenetic relationships of the ciliate class Heterotrichea (Protista, Ciliophora, Postciliodesmatophora) inferred from multiple molecular markers and multifaceted analysis strategy. Mol Phylogenet Evol 78: 118–135 doi: 10.1016/j.ympev.2014.05.012
    Shen YF (1998) The diversity status of freshwater protozoans and the pollution stress on protozoa in China. Biodivers Sci 6: 81 doi: 10.17520/biods.1998012
    Shen YF, Gu MR (2016) Fauna sinica: invertebrata ciliophora oligohymenophorea peritrichida. Science Press, Beijing
    Sheng YL, Pan B, Wei F, Wang YY, Gao S (2021) Case study of the response of N6-Methyladenine DNA modification to environmental stressors in the unicellular eukaryote Tetrahymena thermophila. Msphere 6: e01208-01220
    Simon M, Jardillier L, Deschamps P, Moreira D, Restoux G, Bertolino P, López-García P (2015) Complex communities of small protists and unexpected occurrence of typical marine lineages in shallow freshwater systems. Environ Microbiol 17: 3610–3627 doi: 10.1111/1462-2920.12591
    Slabodnick MM, Ruby JG, Reiff SB, Swart EC, Gosai S, Prabakaran S, Witkowska E, Larue GE, Fisher S, Freeman RM Jr (2017) The macronuclear genome of Stentor coeruleus reveals tiny introns in a giant cell. Curr Biol 27: 569–575 doi: 10.1016/j.cub.2016.12.057
    Small EB, Lynn DH (1985) Phylum Ciliophora Doflein, 1901. In: Lee JJ, Hutner SH, Bovee EC (eds) An illustrated guide to the protozoa. Society of Protozoologists, Lawrence, pp 393–575
    Song WB, Wilbert N (1989) Taxonomische Untersuchungen an Aufwuchsciliaten (Protozoa, Ciliophora) im Poppelsdorfer Weiher, Bonn. Lauterbornia 3: 2–221
    Song WB, Zhao YJ, Xu KD, Hu XZ, Gong J (2003) Pathogenic protozoa in mariculture. Science Press, Beijing
    Song WB, Warren A, Hu XZ (2009) Free-living ciliates in the Bohai and Yellow Seas. Science Press, Beijing
    Song WY, Qiao Y, Dong JY, Bourland WA, Zhang TT, Luo XT (2020) Ontogeny and phylogeny of a new hypotrichous ciliate (Protista, Ciliophora), Metaurostylopsis alrasheidi n. sp., with establishment of a new genus Monourostylopsis n. gen. Front Mar Sci 7: 602317 doi: 10.3389/fmars.2020.602317
    Song W, Xu DP, Chen X, Warren A, Shin MK, Song WB, Li LF (2021a) Overview of the diversity, phylogeny and biogeography of strombidiid oligotrich ciliates (Protista, Ciliophora), with a brief revision and a key to the known genera. Front Microbiol 12: 700940 doi: 10.3389/fmicb.2021.700940
    Song WY, Zhang TY, Dong JY, Luo XT, Bourland WA, Wang YR (2021b) Taxonomy and molecular phylogeny of two new urostylid ciliates (Protozoa: Ciliophora) from Chinese wetlands and establishment of a new genus. Front Microbiol 12: 707954 doi: 10.3389/fmicb.2021.707954
    Strüder-Kypke MC, Wright ADG, Foissner W, Chatzinotas A, Lynn DH (2006) Molecular phylogeny of litostome ciliates (Ciliophora, Litostomatea) with emphasis on free-living haptorian genera. Protist 157: 261–278 doi: 10.1016/j.protis.2006.03.003
    Sun XY, Jiang Z, Liu F, Zhang DZ (2019) Monitoring spatio-temporal dynamics of habitat quality in Nansihu Lake basin, eastern China, from 1980 to 2015. Ecol Indic 102: 716–723 doi: 10.1016/j.ecolind.2019.03.041
    Sun M, Li Y, Cai XL, Liu YQ, Chen Y, Pan XM (2021) Further insights into the phylogeny of peniculid ciliates (Ciliophora, Oligohymenophorea) based on multigene data. Mol Phylogenet Evol 154: 107003 doi: 10.1016/j.ympev.2020.107003
    Vd'ačný P, Foissner W (2012) Monograph of the dileptids (Protista, Ciliophora, Rhynchostomatia). Denisia 31: 1–529
    Vd'ačný P, Foissner W (2013) Synergistic effects of combining morphological and molecular data in resolving the phylogenetic position of Semispathidium (Ciliophora, Haptoria) with description of Semispathidium breviarmatum sp. n. from tropical Africa. Zool Scr 42: 529–549 doi: 10.1111/zsc.12023
    Vd'ačný P, Foissner W (2021) Morphology and ontogenesis of two new Hemiholosticha species (Ciliophora, Hypotrichia, Hemiholostichidae nov. fam.). Eur J Protistol 77: 125763 doi: 10.1016/j.ejop.2020.125763
    Vd'ačný P, Rajter L (2014) An annotated and revised checklist of pleurostome ciliates (Protista: Ciliophora: Litostomatea) from Slovakia, Central Europe. Zootaxa 3760: 501–521 doi: 10.11646/zootaxa.3760.4.1
    Vd'ačný P, Bourland WA, Orsi W, Epstein SS, Foissner W (2011) Phylogeny and classification of the Litostomatea (Protista, Ciliophora), with emphasis on free-living taxa and the 18S rRNA gene. Mol Phylogenet Evol 59: 510–522 doi: 10.1016/j.ympev.2011.02.016
    Vd'ačný P, Rajter L, Shazib SUA, Jang SW, Kim JH, Shin MK (2015) Reconstruction of evolutionary history of pleurostomatid ciliates (Ciliophora, Litostomatea, Haptoria): interplay of morphology and molecules. Acta Protozool 54: 9–29
    Wang CC (1925) Study of the protozoa of Nanking. Part Cont Bio Lab Sci Soc China 1: 1–60
    Wang CD, Gao YY, Lu BR, Chi Y, Zhang TT, El-Serehy HA, Al-Farraj SA, Li LF, Song WB, Gao F (2021a) Large-scale phylogenomic analysis provides new insights into the phylogeny of the class Oligohymenophorea (Protista, Ciliophora) with establishment of a new subclass Urocentria nov. subcl. Mol Phylogenet Evol 159: 107112 doi: 10.1016/j.ympev.2021.107112
    Wang JY, Zhang TT, Li FC, Warren A, Li YB, Shao C (2021b) A new hypotrich ciliate, Oxytricha xianica sp. nov., with notes on the morphology and phylogeny of a Chinese population of Oxytricha auripunctata Blatterer & Foissner, 1988 (Ciliophora, Oxytrichidae). Mar Life Sci Technol 3: 303–312
    Wang JY, Zhao Y, Lu XT, Lyu Z, Warren A, Shao C (2021c) 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). Sci China Life Sci 64: 311–322 doi: 10.1007/s11427-020-1667-3
    Wang Z, Wu T, Lu BR, Chi Y, Zhang X, Al-Farraj SA, Song WB, Warren A, Li LF, Wang CD (2021d) Integrative studies on a new ciliate Campanella sinica n. sp. (Protista, Ciliophora, Peritrichia) based on the morphological and molecular data, with notes on the phylogeny and systematics of the family Epistylididae. Front in Microbiol 12: 718757 doi: 10.3389/fmicb.2021.718757
    Wang YF, Li XH, Chi Y, Song WB, Yan QY, Huang J (2022a) Changes of the freshwater microbial community structure and assembly processes during different sample storage conditions. Microorganisms 10: 1176 doi: 10.3390/microorganisms10061176
    Wang Z, Liu MJ, Ma HG, Lu BR, Shen Z, Mu CJ, Alfarraj SA, El-Serehy HA, Warren A (2022b) Redescription and molecular characterization of two Trichodina species (Ciliophora, Peritrichia, Mobilida) from freshwater fish in China. Parasitol Int 86: 102470 doi: 10.1016/j.parint.2021.102470
    Wang Z, Wu T, Mu CJ, Wang Y, Lu BR, Warren A, Wang CD (2022c) The taxonomy and molecular phylogeny of two epibiotic colonial peritrich ciliates (Ciliophora, Peritrichia). Eur J Protistol 86: 125921 doi: 10.1016/j.ejop.2022.125921
    Wickham H (2016) ggplot2: elegant graphics for data analysis. Springer, New York
    Wilbert N (1975) Eine verbesserte Technik der Protargolimprgnation für Ciliaten. Mikrokosmos 64: 171–179
    Wu T, Li YQ, Lu BR, Shen Z, Song WB, Warren A (2020) Morphology, taxonomy and molecular phylogeny of three marine peritrich ciliates, including two new species: Zoothamnium apoarbuscula n. sp. and Z. apohentscheli n. sp. (Protozoa, Ciliophora, Peritrichia). Mar Life Sci Technol 2: 334–348 doi: 10.1007/s42995-020-00046-y
    Wu L, Li JQ, Warren A, Lin XF (2021a) Morphology and molecular phylogeny of a new brackish pleurostomatid ciliate, Loxophyllum paludosum sp. n. (Ciliophora, Litostomatea, Haptoria), from a mangrove wetland in China. Eur J Protistol 80: 125802 doi: 10.1016/j.ejop.2021.125802
    Wu T, Li YQ, Zhang TT, Hou J, Mu CJ, Warren A, Lu BR (2021b) Morphology and molecular phylogeny of three Epistylis species found in freshwater habitats in China, including the description of E. foissneri n. sp. (Ciliophora, Peritrichia). Eur J Protistol 78: 125767 doi: 10.1016/j.ejop.2021.125767
    Wu T, Wang Z, Duan LL, El-Serehy HA, Al-Farraj SA, Warren A, Liu YJ, Wang CD, Lu BR (2021c) The morphology, taxonomy and phylogenetic analyses of five freshwater colonial peritrich ciliates (Alveolata, Ciliophora), including the descriptions of two new species. Front Microbiol 12: 718821 doi: 10.3389/fmicb.2021.718821
    Wu L, Li JQ, Warren A, Lin XF (2022a) Taxonomy and systematics of a new pleurostomatid ciliate, Pseudolitonotus spirelis gen. et sp. n. (Protozoa, Ciliophora, Haptoria). Mar Life Sci Technol 4: 31–41 doi: 10.1007/s42995-021-00123-w
    Wu T, Liu YJ, Liu Y, Lu BR, Cao X, Hao S, Al-Farraj SA, Warren A, Zhang TT, Wang Z (2022b) Integrative studies on three epibiotic Epistylis species (Protozoa, Ciliophora, Peritrichia) in Lake Weishan Wetland, northern China, including the establishment of a new species and notes on the systematics of sessilids. Protist 173: 125909 doi: 10.1016/j.protis.2022.125909
    Wu T, Wang Z, Lu BR, Lei JT, Al-Rasheid KAS, Sheng YL (2022c) New contribution to the peritrichous genus Ophrydium (Protista, Ciliophora) with notes on the morphology, taxonomy, and phylogeny of a well-known species Ophrydium crassicaule Penard, 1922. J Eukaryot Microbiol 69: e12900
    Xu Y, Gao F, Fan XP (2018) Reconsideration of the systematics of Peniculida (Protista, Ciliophora) based on SSU rRNA gene sequences and new morphological features of Marituja and Disematostoma. Hydrobiologia 806: 313–331 doi: 10.1007/s10750-017-3371-4
    Xu WX, Wang YY, Cheng T, Yu YH, El-Serehy H, Al-Farraj SA, Bourland WA, Luo XT (2020) Reevaluation of the "well-known" Paraurostyla weissei complex, with notes on the ontogenesis of a new Paraurostyla species (Ciliophora, Hypotrichia). Eur J Protistol 73: 125672 doi: 10.1016/j.ejop.2020.125672
    Yan Y, Chen XM, Chen XR, Gao F, Al-Farraj SA, Al-Rasheid KAS (2015) Morphology and molecular phylogeny of three marine Condylostoma species from China, including two new ones (Ciliophora, Heterotrichea). Eur J Protistol 51: 66–78 doi: 10.1016/j.ejop.2014.11.001
    Yan Y, Fan YB, Chen XR, Li LF, Warren A, Al-Farraj SA, Song WB (2016) Taxonomy and phylogeny of three heterotrich ciliates (Protozoa, Ciliophora), with description of a new Blepharisma species. Zool J Linn Soc 177: 320–334
    Zhan ZF, Xu KD, Warren A, Gong YC (2009) Reconsideration of phylogenetic relationships of the subclass Peritrichia (Ciliophora, Oligohymenophorea) based on small subunit ribosomal RNA gene sequences, with the establishment of a new subclass Mobilia Kahl, 1933. J Eukaryot Microbiol 56: 552–558 doi: 10.1111/j.1550-7408.2009.00435.x
    Zhan ZF, Xu KD, Dunthorn M (2013) Evaluating molecular support for and against the monophyly of the Peritrichia and phylogenetic relationships within the Mobilida (Ciliophora, Oligohymenophorea). Zool Scr 42: 213–226 doi: 10.1111/j.1463-6409.2012.00568.x
    Zhang XM, Ji DD, Zhang QQ, Li CH (2015) Description and phylogeny of a new prostomatid, Metacystis similis nov. spec. (Protista, Ciliophora) from the East China Sea. Zootaxa 4033: 584–592 doi: 10.11646/zootaxa.4033.4.8
    Zhang TY, Dong JY, Cheng T, Duan LL, Shao C (2020) Reconsideration of the taxonomy of the marine ciliate Neobakuella aenigmatica Moon et al., 2019 (Protozoa, Ciliophora, Hypotrichia). Mar Life Sci Technol 2: 97–108
    Zhang GAT, Chi Y, Wang Z, Wang Y, Liu R, Warren A, Zhao Y, Pan HB (2022a) Taxonomic and phylogenetic studies on two new freshwater Amphileptus species (Ciliophora, Pleurostomatida), from Lake Weishan, northern China. Eur J Protistol 82: 125854
    Zhang GAT, Zhao Y, Chi Y, Warren A, Pan HB, Song WB (2022b) Updating the phylogeny and taxonomy of pleurostomatid ciliates (Protista, Ciliophora) with establishment of a new family, a new genus and two new species. Zool J Linn Soc 196: 105–123
    Zhao XL, Li Y, Duan LL, Chen X, Mao FB, Juma M, Liu YF, Song WB, Gao S (2020) Functional analysis of the methyltransferase SMYD in the single-cell model organism Tetrahymena thermophila. Mar Life Sci Technol 2: 109–122
  • 加载中
通讯作者: 陈斌, bchen63@163.com
  • 1. 

    沈阳化工大学材料科学与工程学院 沈阳 110142

  1. 本站搜索
  2. 百度学术搜索
  3. 万方数据库搜索
  4. CNKI搜索

Figures(10) / Tables(7)

Article Metrics

Article views(42) PDF downloads(0) Cited by()

Related
Proportional views

Biodiversity of freshwater ciliates (Protista, Ciliophora) in the Lake Weishan Wetland, China: the state of the art

    Corresponding author: Borong Lu, boronglu@126.com
  • 1. Institute of Evolution and Marine Biodiversity, Ocean University of China, Qingdao, 266003, China
  • 2. Marine College, Shandong University, Weihai, 264209, China
  • 3. Laoshan Laboratory, Qingdao, 266237, China
  • 4. Zoology Department, College of Science, King Saud University, Riyadh, 11451, Saudi Arabia
  • 5. Department of Life Sciences, Natural History Museum, London, SW7 5BD, UK

Abstract: Ciliates are core components of the structure of and function of aquatic microbial food webs. They play an essential role in the energy flow and material circulation within aquatic ecosystems. However, studies on the taxonomy and biodiversity of freshwater ciliates, especially those in wetlands in China are limited. To address this issue, a project to investigate the freshwater ciliates of the Lake Weishan Wetland, Shandong Province, commenced in 2019. Here, we summarize our findings to date on the diversity of ciliates. A total of 187 ciliate species have been found, 94 of which are identified to species-level, 87 to genus-level, and six to family-level. These species show a high morphological diversity and represent five classes, i.e., Heterotrichea, Litostomatea, Prostomatea, Oligohymenophorea, and Spirotrichea. The largest number of species documented are oligohymenophoreans. A comprehensive database of these ciliates, including morphological data, gene sequences, microscope slide specimens and a DNA bank, has been established. In the present study, we provide an annotated checklist of retrieved ciliates as well as information on the sequences of published species. Most of these species are recorded in China for the first time and more than 20% are tentatively identified as new to science. Additionally, an investigation of environmental DNA revealed that the ciliate species diversity in Lake Weishan Wetland is higher than previously supposed.

    HTML

Introduction
  • Ciliates (phylum Ciliophora Doflein, 1901) are a large monophyletic group of unicellular eukaryotes that are the subject of investigation in numerous biological disciplines, such as systematics, evolution, ecology, and cytology, due to their unique cellular structure, functional ecology and life cycles (Gao et al. 2016; Huang et al. 2021; Liu et al. 2021; Song et al. 2021a; Wang et al. 2021a; Zhao et al. 2020). Species of genera, such as Tetrahymena, Paramecium, and Stentor, have been successfully cultured in the laboratory as model organisms to investigate a wide range of biological processes and issues (Sheng et al. 2021; Slabodnick et al. 2017). Nevertheless, knowledge and understanding of ciliate taxonomy and biodiversity, which are important branches of systematics and underpin numerous related disciplines, are limited and it has been estimated that 83–89% of free-living ciliate species have yet to be discovered (Foissner et al. 2008a, b).

    In China, the study of ciliates began in 1925 when Wang reported dozens of species (Wang 1925). From then to the 1990s, researchers investigated ciliates from various freshwater habitats in more than 20 provinces in China (Institute of Hydrobiology Chinese Academy of Sciences 1973; Shen 1998; Shen and Gu 2016). However, due to technical limitations, key morphological data, including descriptions of taxonomically important characters such as the pattern of the ciliature, and clear photomicrographs are lacking for most freshwater species reported during this period. Since the 1990s, studies on the taxonomy and systematics of ciliates have been based on modern techniques and, as a result, significant advances have been made in knowledge and understanding of the biodiversity and evolutionary relationships of ciliates in China. However, these works are highly unbalanced in the sense that most have focused on marine rather than freshwater ciliates. This is exemplified by the extensive faunal surveys of the peninsula of Shandong Province and of the coastline of southern China, carried out by several research groups including the Laboratory of Protozoology at the Ocean University of China (Hu et al. 2019; Song et al. 2003, 2009). These works include diversity data for more than 900 species, supported by detailed descriptions of the morphology, phylogeny, genomics and gene evolution, and ecology of many of these. In contrast, there have been only sporadic studies on ciliates from freshwater habitats. To address this deficiency, it is necessary to conduct long-term systematic studies on freshwater ciliate diversity using modern techniques.

    Lake Weishan Wetland (LWW) stretches from south to north in a long narrow strip in the southwestern part of Shandong Province, China, and has a drainage area of 31, 700 km2 (Fig. 1). Because of the effects of natural and artificial factors, it has a variety of ecological and environmental habitats resulting in a rich biodiversity (Sun et al. 2019). In addition, as an important node in the South-North Water Diversion Project in China, the biodiversity and ecosystem health of this area have increasingly been a focus of attention. Therefore, we selected the LWW to carry out a 5-year (2019–2023) project on the diversity of freshwater ciliates.

    Figure 1.  Sampling sites and habitat characteristics in Lake Weishan Wetland. The insert figure of partial part of China (upper right) indicates the location of Lake Weishan Wetland

    About 200 ciliate species have been found during the first phase of the project, running from 2019 to 2021. A comprehensive database including morphological information, localities, ecological data, gene sequences, microscope slide specimens and a DNA bank, has been established for these species. The morphological information mainly comprises detailed descriptions, photomicrographs and video sequences of living specimens, images of silver-stained specimens, and hand-drawn illustrations. Here, we provided an annotated checklist of retrieved organisms to show the preliminary findings of our investigation. Most of these species are new records for China and about 40 are new to science. The taxonomy of many species has or will be refined or revised. Several publications have been produced and others are in preparation (for details, see Tables 1, 2, 3, 4, 5, 6 and 7). Furthermore, an investigation of environmental DNA revealed that there is a large undiscovered ciliate diversity in the LWW. Here, we provide an overview of the findings of the project to date.

    Family Genus Species Details on sampling Publication GenBank Acc. No.
    Blepharismidae Blepharisma Blepharisma penardi Lake
    Blepharisma undulans Ditch
    Blepharisma sp. (undescr. sp.?) Lake
    Climacostomidae Climacostomum Climacostomum virens Lake
    Condylostomatidae Condylostomides Condylostomides coeruleus River
    Linostomella Linostomella pseudovorticella Channel
    Spirostomidae Spirostomum Spirostomum caudatum River Chi et al. (2022a) OK274292;
    OM127346;
    OM127339
    Stentoridae Stentor Stentor coeruleus Fish pond
    Stentor roeselli Fish pond
    Stentor sp. Lake
    –, data not available
    undescr. sp., undescribed species (possibly new species); GenBank Acc. No., GenBank Accession Number

    Table 1.  Taxa of heterotrichs investigated in the present study

    Order Family Genus Species Details on sampling Publication GenBank Acc. No.
    Cyclotrichiida Mesodiniidae Askenasia Askenasia volvox Fish pond
    Haptorida sensu Lynn (2008) Acropisthiidae Balantidion 1 Balantidion foissneri (undescr. sp.)2 Channel Chi et al. (2022b) ON158511
    Balantidion pellucidum Fish pond
    Undefined3 undescr. gen. & sp.? Fish pond
    Undefined3 undescr. gen. & sp.? Fish pond
    Actinobolinidae Actinobolina Actinobolina radians Fish pond
    Actinobolina sp. (undescr. sp.) Fish pond
    Didiniidae Didinium Didinium nasutum Lake
    Monodinium Monodinium sp. Lake
    Dileptidae Apodileptus Apodileptus visscheri Lake
    Paradileptus Paradileptus elephantinus Pontoon dock Chi et al. (2021b) MZ147012; MZ574467
    Paradileptus conicus Fish pond Chi et al. (2021b) MZ147013; MZ574468
    Pseudomonilicaryon Pseudomonilicaryon fraterculum Lake
    Enchelyidae Enchelys Enchelys sp. Fish pond
    Papillorhabdos Papillorhabdos sp. (undescr. sp.) Fish pond
    Enchelyodontidae Enchelydium Enchelydium sp. Fish pond
    Fuscheriidae Fuscheria Fuscheria sp. (undescr. sp.?) Lake
    Homalozoonidae Homalozoon Homalozoon vermiculare River
    Kamburophryidae Kamburophrys Kamburophrys sp. Ditch
    Lacrymariidae Lacrymaria Lacrymaria olor Fish pond
    Lacrymaria sp. 1 (undescr. sp.?) Lake
    Lacrymaria sp. 2 (undescr. sp.?) Lake
    Myriokaryonidae Myriokaryon Myriokaryon lieberkuehnii Lake
    Spathidiidae Neobryophyllum Neobryophyllum sp. (undescr. sp.?) Fish pond
    Spathidium Spathidium polymophum? Fish pond
    Tracheliidae Trachelius Trachelius ovum Ditch
    Trachelophyllidae Enchelyotricha Enchelyotricha sp. 1 (undescr. sp.?) Lake
    Enchelyotricha sp. 2 (undescr. sp.?) Lake
    Enchelyotricha sp. 3 Channel
    Foissnerides Foissnerides sp. Fish pond
    Lagynophrya Lagynophrya sp. (undescr. sp.?) River
    Trachelophyllum Trachelophyllum brachypharynx Lake
    undescr. sp. undescribed species (may be new species), GenBank Acc. No. GenBank Accession Number
    1Family assignation according to Chi et al. 2022a, b
    2New published species
    3Same genus
    –, data not available

    Table 2.  Taxa of haptorids s. lat. investigated in the present study

    Family Genus Species Details on sampling Publication GenBank Acc. No.
    Amphileptidae Amphileptus Amphileptus parapleurosigma 1 Wetland Zhang et al. (2022a) MW784241
    Amphileptus weishanensis 1 Port Zhang et al. (2022a) MW784242
    Amphileptus sp. 1 (undescr. sp.)2 Port OL828281
    Amphileptus sp. 2 (undescr. sp.)2 Fish pond OL828282
    Amphileptus sp. 3 (undescr. sp.)2 Ditch OL828283
    Amphileptus sp. 4 Wetland
    Amphileptus sp. 5 Fish pond
    Amphileptus sp. 6 Fish pond
    Pseudoamphileptus Pseudoamphileptus sp. Host: Cyprinus carpio; fish pond
    Litonotidae Litonotus Litonotus pygmaeus Wetland
    Litonotus sp. 1 Fish pond
    Litonotus sp. 2 Lake
    Litonotus sp. 3 Fish pond
    Loxophyllum Loxophyllum apochlorelligerum 1 Wetland Zhang et al. (2022b) MW414675
    Loxophyllum sp. 1 Wetland
    Loxophyllum sp. 2 Wetland
    Protolitonotidae Protolitonotus Protolitonotus sp. 1 Lake
    undescr. sp. undescribed species (may be new species), GenBank Acc. No. GenBank Accession Number
    1New published family or genus or species
    2Have been submitted for publication
    –, data not available

    Table 3.  Taxa of pleurostomatids investigated in the present study

    Order Family Genus Species Details on sampling
    Prorodontida Colepidae Coleps Coleps elongatus Lake
    Holophryidae Holophrya Holophrya discolor An anoxic pool
    Holophrya teres Ditch
    Holophrya sp. Lake
    Lagynidae Lagynus Lagynus sp. An anoxic pool
    Urotrichidae Bursellopsis Bursellopsis sp. Lake
    Urotricha Urotricha valida Fish pond
    Urotricha sp. Lake
    Prostomatida Apsiktratidae Apsiktrata Apsiktrata sp. An anoxic pool

    Table 4.  Taxa of prostomateans investigated in the present study (no publication and no molecular data of these prostomateans online so far)

    Order Family Genus Species Details on sampling Publication GenBank Acc. No.
    Loxocephalida Conchophthiridae Conchophthirus Conchophthirus sp. 1 Host: Sinanodonta woodiana; wetland
    Conchophthirus sp. 2 Host: Cristaria plicata; wetland
    Loxocephalidae Dexiotricha Dexiotricha granulosa wetland
    Philasterida Parauronematidae Glauconema Glauconema sp. (undescr. sp.) Wetland
    Uronematidae Uronema Uronema nigricans Aquaculture pond
    Uronema sp. Wetland
    Urozonidae Urozona Urozona sp. Ditch
    Pleuronematida Cyclidiidae Cyclidium Cyclidium sp. 1 River
    Cyclidium sp. 2 River
    Cyclidium sp. 3 Aquaculture pond
    Histiobalantiidae Histiobalantium Histiobalantium sp. Wetland
    Pleuronematidae Pleuronema Pleuronema sp. (undescr. sp.) Wetland
    Thigmophryidae Myxophyllum Myxophyllum sp. (undescr. sp.) Host: Acusta ravida; wetland
    Peniculida Frontoniidae Frontonia Frontonia atra River Li et al. (2022) MZ437443
    Frontonia apoelegans River Li et al. (2022) MZ437442
    Frontonia sp. 1 (undescr. sp.) Aquaculture pond
    Frontonia sp. 2 Pond
    Lembadionidae Lembadion Lembadion lucens River
    Lembadion magnum Wetland
    Lembadion bullinum River
    Parameciidae Paramecium Paramecium caudatum Pond
    Paramecium sp. Pond
    Stokesiidae Marituja Marituja sp. (undescr. sp.) River
    Stokesia Stokesia vernalis River Li et al. (2022) MZ437444
    undescr. sp. undescribed species (may be new species), GenBank Acc. No. GenBank Accession Number
    –, data not available

    Table 5.  Taxa of scuticociliates and peniculids investigated in the present study

    Order Family Genus Species Details on sampling Publication GenBank Acc. No.
    Sessilida Astylozoidae Hastatella Hastatella sp. Aquaculture pond
    Undefined undescr. gen. & sp.? Aquaculture pond
    Epistylididae Campanella Campanella sinica1 Ditch Wang et al. (2021d) MW969624; MW969625; MW969627
    Epistylis Epistylis anastatica Host: cyclopoid copepods; aquaculture pond
    Epistylis cambari Host: Procambarus clarkia; aquaculture pond
    Epistylis chlorelligerum Wetland
    Epistylis daphniae Host: Daphnia sp; ditch Wu et al. (2022b) OP289655
    Epistylis foissneri1 Wetland Wu et al. (2021b) MW172838
    Epistylis galea Wetland
    Epistylis hentscheli Aquaculture pond Wu et al. (2021b) MW172839
    Epistylis lwoffi2 Host: Ctenopharyngodon idella; aquaculture pond
    Epistylis pygmaeum Host: Rotifera; aquaculture pond Wu et al. (2022b) OP289656
    Epistylis semiciculus Host: Procambarus clarkia; aquaculture pond
    Epistylis tubificis Wetland
    Epistylis vaginula Wetland Wu et al. (2021b) MW172840
    Epistylis weishanensis1 Host: Gammarus sp; wetland Wu et al. (2022b) OP289654
    Epistylis sp. 1 Host: Procambarus clarkia; aquaculture pond
    Epistylis sp. 2 Wetland
    Epistylis sp. 3 Wetland
    Orborhabdostyla Orborhabdostyla sp. (undescr. sp.)3 Aquaculture pond
    Operculariidae Opercularia Opercularia nutans Float grass in a lake
    Undefined Undefined Aquaculture pond
    Ophrydiidae Ophrydium Ophrydium crassicaule Float grass in a lake Wu et al. (2022c) OM801561
    OM801562
    OM801564
    Scyphidiidae Undefined Undescr. gen. & sp.3 Host: Ctenopharyngodon idella; aquaculture pond
    Vaginicolidae Cothurnia Cothurnia sp. Fish pond
    Pyxicola Pyxicola carteri Fish pond
    Vorticellidae Carchesium Carchesium cyclopidarum Ditch Wang et al. (2022c) ON158510
    Carchesium polypinum Fish pond Wu et al. (2021c) MZ478146
    Epicarchesium Epicarchesium granulatum Host: Procambarus clarkia; aquaculture pond
    Epicarchesium sinense1 Aquaculture pond Wu et al. (2021c) MZ478145
    Pseudovorticella Pseudovorticella sp. 1 Fish pond
    Pseudovorticella sp. 2 Pond
    Pseudovorticella sp. 3 (undescr. sp.?) Ditch
    Vorticella Vorticella sp. 1 Pond
    Vorticella sp. 2 Pond
    Vorticella sp. 3 Fish pond
    Vorticella sp. 5 Float grass in a lake
    Vorticella sp. 6 Wetland
    Vorticella sp. 7 Fish pond
    Vorticella sp. 8 Wetland
    Vorticella sp. 9 Ditch
    Vorticella sp. 10 Port
    Vorticella sp. 11 (undescr. sp.?) Float grass a lake
    Vorticellides Vorticellides aquadulcis Aquarium
    Zoothamniidae Zoothamnium Zoothamnium arbuscula Wetland Wu et al. (2021c) MZ478144
    Zoothamnium hentscheli Wetland Wu et al. (2021c) MZ478143
    Zoothamnium procerius Host: Procambarus clarkia; aquaculture pond
    Zoothamnium weishanicum1 Wetland Wu et al. (2021c) MZ478142
    Zoothamnium sp. Wetland
    Mobilida Trichodinidae Trichodina Trichodina acuta Host: Cyprinus carpio; fish pond Wang et al. (2022b) MT982920
    Trichodina funduli Host: Cyprinus carpio; fish pond
    Trichodina nigra Host: Cyprinus carpio; fish pond Wang et al. (2022b) MT982921
    Trichodina nobillis Host: Ctenopharyngodon idella; fish pond
    Trichodina pediculus Host: Hydra sp.; ditch
    Trichodina sp. 1 Host: Channa argus; fish pond
    Trichodina sp. 2 Host: Siniperca chuatsi; fish pond
    Trichodina sp. 3 Host: Anodonta woodiana; fish pond
    Trichodinella Trichodinella sp. Host: Ctenopharyngodon idella; fish pond
    undescr. sp. undescribed species (new to science), GenBank Acc. No. GenBank Accession Number
    1New published species
    2Transferred into genus Heteropolaria by Foissner and Schubert (1977), however, reassigned into Epistylis by Wu et al. (in preparation)
    3In preparation for publication
    –, data not available

    Table 6.  Taxa of peritrichs investigated in the present study

    Order/group Family Genus Species Details on sample Publication GenBank Acc. No.
    Dorsomarginalia Bothrigidae Bothrix Bothrix africana Fish pond
    Uroleptidae Uroleptus Uroleptus gallina Wetland
    Uroleptus longicaudatus Wetland
    Uroleptus sp. Wetland
    Sporadotrichida Keronidae Kerona Kerona pediculus Host: Hydra sp.; ditch
    Oxytrichidae Cyrtohymena Cyrtohymena primicirrata Ditch
    Monomicrocaryon Monomicrocaryon euglenivorum euglenivorum Wetland
    Notohymena Notohymena australis Ditch
    Oxytricha Oxytricha sp. 1 Wetland
    Oxytricha sp. 2 Wetland
    Paraurostyla Paraurostyla weissei Wetland
    Pleurotricha Pleurotricha curdsi Wetland
    Pseudostromhidium Pseudostromhidium planctonticum Fish pond
    Rubrioxytricha Rubrioxytricha haematoplasma Wetland
    Sterkiella Sterkiella sp. Wetland
    Stylonychia Stylonychia pustulata Wetland
    Urosoma Urosoma caudata Ditch
    Stichotrichida Chaetospiridae Chaetospira Chaetospira sp. (undescr. sp.) Fish pond OM313318;
    OM313319;
    OM313320
    Stichotricha Stichotricha sp. Wetland
    Deviatidae Deviata Deviata polycirrata Brook
    Undefined undescr. gen. & sp.? Brook
    Spirofilidae Hypotrichidium Hypotrichidium sp. Ditch
    Strongylidiidae Pseudouroleptus Pseudouroleptus sp. 1 (undescr. sp.) Fish pond
    Pseudouroleptus sp. 2 Fish pond
    Strongylidium Strongylidium wuhanense Wetland
    Urostylida Bakuellidae Heterobakuella1 Heterobakuella bergeri 1 Brook Song et al. (2021b) MW692986
    Pseudokeronopsidae Pseudokeronopsis Pseudokeronopsis erythrina Lake
    Pseudourostylidae Diaxonella Diaxonella trimarginata Wetland
    Urostylidae Holosticha Holosticha pullaster Ditch
    Neourostylopsis Neourostylopsis flava Wetland
    Metaurostylopsis Metaurostylopsis alrasheidi1 Wetland Song et al. (2020) MT911525
    Pseudourostla Pseudourostla cristata Ditch
    Euplotida Euplotidae Euplotes Euplotes woodruffi Wetland
    Euplotes muscicola Fish pond
    Euplotes sp. 1 Wetland
    Euplotes sp. 2 Wetland
    Euplotes sp. 3 Wetland
    undescr. sp undescribed species (may be new species), GenBank Acc. No. GenBank Accession Number
    1New published genus or species
    –, data not available

    Table 7.  Taxa of hypotrichs s. lat. investigated in the present study

Results and discussion

    Ecological information (Fig. 2)

  • Figure 2.  A The variations of surface water temperature, electrical conductivity (EC) and OTU richness of Ciliophora (plankton community) from August 2020 to July 2021. B The variations of nitrate (NO3), pH and dissolved oxygen concentration (DO) from August 2020 to July 2021. C Temporal variations in the plankton community composition of Ciliophora based on the rank of order from August 2020 to July 2021

    From August 2020 to July 2021, a total of 12 water samples and corresponding physicochemical parameters were obtained from LWW. The monthly trends of physicochemical parameters and OTU richness of the planktonic ciliate community are as follows.

  • Surface water temperature
  • The surface water temperature decreased from 31.7 to 3.0 ℃ during the months August to December, and then increased from 3.0 to 29.2 ℃ during the months January to June (Fig. 2A).

  • Electrical conductivity (EC)
  • The EC declined rapidly from 1015 to 571 μS/cm from August to January but increased gradually from 571 to 673 μS/cm during January to March, and then increased rapidly from 673 to 1065 μS/cm during May to July (Fig. 2A).

  • Dissolved oxygen concentration (DO)
  • The DO increased from 5.06 to 16.03 mg/L during the months August to January and then decreased from 16.03 to 5.29 mg/L during the months January to July (Fig. 2B).

  • pH
  • The pH increased from 8.05 in August to 8.44 in September, remained almost stable from September to April, and then increased slightly in May (8.84) and June (8.82). The minima and maxima of pH were in August (8.05) and May (8.84), respectively (Fig. 2B).

  • Nitrate (NO3)
  • The NO3 concentration increased from 0.17 to 4.85 mg/L from August to September and then declined to 0.14 mg/L in October. It remained at a low level from October to December, increased from 0.19 to 16.55 mg/L during December to February, and then decreased again from 16.55 to 0.18 mg/L during February to July (Fig. 2B).

  • Operational taxonomic units (OTUs)
  • In total, 30, 076 high-quality sequences of Ciliophora (plankton community) were clustered into 273 OTUs at the 97% similarity level and were assigned to 31 genera, nine families, three orders and one class based on the PR2 database (Supplementary Table S1). The OTU richness exhibited a negative correlation with temperature (r = -0.63, p < 0.05) and EC (r = -0.60, p < 0.05) but was positively correlated with DO (r = 0.62, p < 0.05) (Fig. 2). The OTU richness increased from 26 to 102 during August to January, declined to 36 in March, increased to 61 in April and then decreased again to 27 during April to July (Fig. 2A).

    Based on the sequence data, the planktonic ciliate community was dominated by the Spirotrichea Bütschli, 1889 (79.40%) and also included CONthreeP (20.56%) and Litostomatea Small & Lynn, 1981 (0.04%) from August 2020 to July 2021 (Fig. 2C). Spirotrichea were dominant in all months except November, when CONthreeP (Colpodea, Oligohymenophorea, Nassophorea, Phyllopharyngea, Plagiopylea and Prostomatea) was the most representative group. Litostomatea appeared in only a few months (Fig. 2C). Tintinnidium was the most abundant genus, accounting for 30.34% of rRNA gene relative abundance in the ciliate communities, followed by Halteria (13.33%).

  • Overview of species diversity based on morphological observations (Fig. 3)

  • Figure 3.  The species distributions of the investigated groups. A The investigated groups. The leaves represent the families included in each group, and the different sizes of leaves represent different values of species number in each family. B, C Species distributions at order-level. D species distributions at family-level

    A total of 187 ciliate morphospecies have been found including 94 identified to species-level, 87 to genus-level, and six species to family-level. The class Oligohymenophorea de Puytorac et al., 1974 was represented by the largest number of species (82), followed by Litostomatea (49), Spirotrichea (37), Heterotrichea Stein, 1859 (10), and Prostomatea Schewiakoff, 1896 (9). The two top-ranked groups at the order-level were Sessilida Kahl, 1933 and Haptorida Corliss, 1974 sensu Lynn (2008) with 49 and 31 species, respectively, while Cyclotrichiida Jankowski, 1980 and Prostomatida Schewiakoff, 1896 only had one species each. At the family-level, Vorticellidae Ehrenberg, 1838 and Epistylididae Kahl, 1933 had the largest number of species, with 18 of each. Thirteen articles have been published with details of these findings (Chi et al. 2021b, 2022a, b; Li et al. 2022; Wang et al. 2021d, 2022b, c; Wu et al. 2021b, c, 2022b, c; Zhang et al. 2022a, b) and several others are in preparation.

  • Species diversity of heterotrichs (Fig. 4; Table 1)

  • Figure 4.  Selected heterotrichous ciliates found in Lake Weishan Wetland (original). A Spirostomum teres, arrowhead indicates the contractile vacuole, arrow indicates the macronucleus. B Stentor coeruleus. C Climacostomum virens, arrowheads indicate the cyrtos, arrow indicates the contractile vacuole. D, E Blepharisma sp., arrowheads indicate the cortical granules, arrows indicate the paroral membrane, double arrowheads indicate the adoral zone of membranelles. F Linostomella pseudovorticella. G Oral infraciliature of Stentor coeruleus. H Stentor spp., arrowheads indicate Stentor roeselii, arrows indicate Stentor coeruleus. I Stentor roeselii, arrow indicates the contractile vacuole. J Cortical granules (arrowheads) of Climacostomum virens. K Cortical granules (arrowheads) of Stentor roeselii. L Cortical granules (arrowheads) of Stentor coeruleus. M General infraciliature of Stentor coeruleus adjusted by the invert function via Photoshop. Ma Macronucleus. Scale bars 50 μm (C), 100 μm (A, D, F, I), 300 μm (B, H, M)

  • Background
  • The class Heterotrichea is considered to be a primitive group within the Ciliophora. Heterotricheans are generally characterized by their large body size, somatic dikinetids associated with postciliodesmata, and an oral apparatus comprising a paroral membrane and an adoral zone of membranelles (Chi et al. 2021a). Due to their large size and omnivorous mode of nutrition, they usually feed on various organisms, from bacteria to small metazoans and microalgae (Foissner and Berger 1996). According to the most recent classification of heterotrichs (Shazib et al. 2014), the class Heterotrichea contains ten families and 58 genera. In China, biodiversity studies on heterotrichs were scarce until the last decade of the twentieth century, and few freshwater species have been systematically investigated (Chi et al. 2020; Hu et al. 2019; Song et al. 2009; Yan et al. 2015, 2016).

  • Results and remarks
  • To date, ten species belonging to five families and six genera have been investigated in detail using morphological and molecular methods (Fig. 4; Table 1). Details of one species, i.e., Spirostomum caudatum (Müller, 1786) Delphy, 1939 have been published (Chi et al. 2022a). One species of Blepharisma is suspected as new based on comparisons with its congeners. In addition, Condylostomides coeruleus Foissner, 2016, which has long been regarded as a biogeographically restricted flagship species and hitherto known only from North America (Hines et al. 2020), was recovered from LWW.

  • Species biodiversity of haptorids (Fig. 5AH, JL; Table 2)

  • Figure 5.  Selected haptorid (AH, JL) and prorodontid (I, MP) ciliates found in Lake Weishan Wetland (original). A, B Pseudomonilicaryon spp., arrowheads indicate the proboscis, arrow indicates the oral opening. C, J, K Enchelyotricha spp., arrow in J indicates the contractile vacuole, arrow in K indicates the oral bulge. D, E Paradileptus sp., arrows in D indicate the contractile vacuoles, arrow in E indicates the oral opening, arrowhead in E indicates the proboscis. F Trachelius ovum, arrow indicates the proboscis. G Didinium sp. H Homalozoon sp. I, O, P Holophrya sp. after protargol staining (I), in vivo (O), and after dry silver nitrate staining (P), arrow indicates the brosse, arrowhead indicates the oral opening. L Kamburophrys sp. M, N Coleps sp. in vivo (M) and after protargol staining (N), arrows indicate the spines. Scale bars 60 μm

  • Background
  • Haptorids are a highly diverse group with more than 1000 nominal species. They show significant variation in size and shape, whereas typical haptorids have an apically or laterally located mouth (Kahl 1930; Lynn 2008). They are raptorial ciliates that feed on flagellates, other ciliates, and even metazoans, such as rotifers and nematodes (Foissner et al. 1995, 1999; Lynn 2008). Although the classification of haptorids has been modified dramatically during the past half-century, taxonomic chaos prevails in much of this group (Huang et al. 2018; Strüder-Kypke et al. 2006; Vd'ačný and Foissner 2013; Vd'ačný and Rajter 2014; Vd'ačný et al. 2011). Recent studies have shown that the diversity of haptorids is much greater than had been previously assumed (Foissner and Oertel 2009; Kwon et al. 2014; Vd'ačný and Foissner 2012). Hitherto, more than 50 species from freshwater habitats, and fewer than 30 species from marine and brackish habitats, have been recorded in China since Wang (1925) described 11 haptorids from Nanjing (Chiang et al. 1983; Hu et al. 2019; Jiang et al. 2021; Shen 1998; Song et al. 2009). However, data, such as photomicrographs, infraciliature, morphometry and details of many key characters, are lacking for many of these species. Consequently, knowledge of the biodiversity of haptorids in China is still in its infancy.

  • Results and remarks
  • In the past three years, we investigated 31 haptorids belonging to 14 families isolated from the LWW (Table 2). At least ten of these 31 haptorids have been reported from other continents, supporting their wide distribution model. More than ten unidentified species may be new to science. The generic assignments of two unidentified species of Acropisthiidae Foissner & Foissner, 1988 are uncertain and may represent a new genus. In addition to the type species of Balantidion, B. pellucidum Eberhard, 1862, another species of this previously monotypic genus was described, namely Balantidion foissneri Chi et al., 2022. Based on the morphology of these two Balantidion species, Chi et al. (2022b) agreed with Foissner et al. (1999) that Balantidion should be assigned to the family Acropisthiidae Foissner & Foissner, 1988 rather than Enchelyidae Ehrenberg, 1838 as suggested by Lynn (2008). Details of two insufficiently known species of Paradileptus have been published by Chi et al. (2021b). Some rare species, i.e., found only once or twice during the last half-century, were isolated from LWW, such as Kamburophrys sp.

  • Species diversity of pleurostomatids (Fig. 6; Table 3)

  • Figure 6.  Selected pleurostomatid ciliates found in Lake Weishan Wetland. A Amphileptus sp. B–E Kentrophyllum spp., white arrows indicate the macronuclear nodules, red arrows indicate the contractile vacuoles. F Loxophyllum sp. G, N, P Amphileptus weishanensis, arrows in (P) indicate the macronuclear nodules. H, I Amphileptus sp., white arrows indicate the macronuclear nodules. J, L Protolitonotus sp. K Litonotus sp. M Amphileptus parapleurosigma. O Amphileptus sp., red arrow marks the extrusomes. Scale bars 100 μm (A, H, I, J, L–N), 75 μm (B–F), 200 μm (G, P), 50 μm (K), 60 μm (O)

  • Background
  • The order Pleurostomatida Schewiakoff, 1896 is a group of raptorial ciliates comprising nearly 200 nominal species. Pleurostomatids are characterized by the bilaterally compressed body, the oral region located along the ventral margin and the bristles on the left side (Foissner et al. 1995; Kahl 1931, 1933; Lynn 2008; Song and Wilbert 1989). Since most pleurostomatids share a similar body shape, it is difficult to distinguish them based only on observations of cells in vivo. Recent studies based on a combination of morphological and molecular data have revealed a high pleurostomatid diversity and resolved numerous taxonomic confusions (Pan et al. 2020; Vd'ačný et al. 2015; Wu et al. 2021a, 2022a; Zhang et al. 2022a, b). However, phylogenetic relationships within this group are far from being resolved entirely, mainly due to undersampling and the paucity of gene sequence data.

  • Results and remarks
  • To date, we have identified 17 pleurostomatids belonging to three families and five genera from the LWW (Table 3). Three new species, Amphileptus weishanensis Zhang et al., 2022, Amphileptus parapleurosigma Zhang et al., 2022, and Loxophyllum apochlorelligerum Zhang et al., 2022, have been described by Zhang et al.(2022a, b). Some poorly known species and genera were also found in the LWW, such as Litonotus pygmaeus Vuxanovici, 1959 and Pseudoamphileptus Foissner, 1983, all of which lacked reliable morphological data and are being re-investigated based on detailed integrative studies.

  • Species diversity of prostomateans (Fig. 5I, MP; Table 4)

    Background
  • Prostomateans are common members of the planktonic community where they can achieve significant abundance and have therefore attracted wide attention from ecologists (Esteban and Fenchel 2021; Foissner et al. 1999; Frantal et al. 2022; Pröschold et al. 2021). In Lynn's (2008) classification, prostomateans comprise two orders, Prostomatida and Prorodontida Corliss, 1974. Prostomatids are characterized by having a genuinely apical oral region, perioral kineties that form obvious paratenes, and neither a brosse nor toxicysts. Prorodontids have an apical or slightly subapical oral region, a brosse and toxicysts. Many prostomateans have not been investigated using modern techniques, even though silver-staining is indispensable for correctly identifying most species (Foissner 2012, 2021; Foissner and Pfister 1997; Foissner et al. 2008a, b; Frantal et al. 2022). In the last decade, the number of new taxa of prostomateans has increased slowly. However, the findings of Jiang et al.(2021, 2022) suggest that this paucity of species is likely due to undersampling.

  • Results and remarks
  • We have investigated nine prostomateans representing five families and six genera (Table 4) and extracted and stored DNA from each to acquire molecular data for future phylogenetic analyses. No findings of our studies on prostomateans have yet been published and investigations of several isolates are ongoing. Nevertheless, new morphological and molecular data are expected to become available for members of the poorly known genera Lagynus, Bursellopsis and Apsiktrata. Up to now, detailed morphological and molecular data are scarce for prostomateans. Molecular data (SSU rDNA) are available for only two prostomatids, i.e., Metacystis similis Zhang et al., 2015 and Foissnerophrys alveolata Jiang et al., 2021. Phylogenetic analyses of these two questioned the systematics of Prostomatida since they do not form a clade, i.e., Metacystis similis is sister to Holophryidae within the other order Prorodontida, and Foissnerophrys alveolata groups with the plagiopyleans (Jiang et al. 2021; Zhang et al. 2015). Future studies will focus on the systematics of prostomateans.

  • Species diversity of scuticociliates and peniculids (Fig. 7; Table 5)

  • Figure 7.  Selected peniculids and scuticociliates found in Lake Weishan Wetland (original). A Paramecium sp., arrow indicates the caudal cilia. B Frontonia sp., arrow indicates the buccal cavity, arrowhead indicates the postoral suture. C Myxophyllum sp. D, E Glauconema sp. after silver nitrate (D) and protargol staining (E), arrows in D indicate the contractile vacuole pores, arrow in E indicates the caudal complex. F Marituja sp. G Pleuronema sp., arrow indicates the paroral membrane. H Dexiotricha granulosa, arrow indicates the caudal cilium. I, J, N Lembadion lucens in vivo (I), after silver nitrate staining (J) and after protargol staining (N), arrow in (I) indicates the buccal cavity, arrow in (N) indicates macronucleus. K Cyclidium sp. L, M Uronema nigricans, in vivo (L) and after wet silver nitrate staining (M, image processed by the invert function via Photoshop). O Lembadion bullinum. Abbreviation: Ma, Macronucleus. Scale bars 100 μm (A, B, F), 40 μm (C), 30 μm (G, I, J, N), 20 μm (H), 10 μm (K–M), 70 μm (O)

  • Background
  • Members of the subclasses Scuticociliatia Small, 1967 and Peniculia Fauré-Fremiet in Corliss, 1956 are extremely widespread in freshwater, brackish and marine habitats (Jung 2021; Li et al. 2021; Liu et al. 2022; Poláková et al. 2021; Serra et al. 2021a, b; Sun et al. 2021; Xu et al. 2018). In these groups, the oral apparatus is generally composed of three membranelles (scuticociliatids) or peniculi (peniculids) and one paroral membrane. The subclass Scuticociliatia comprises more than 300 nominal species. Scuticociliates usually swim fast and are small in size (length about 15–50 μm). While most are free-living and tend to multiply rapidly in eutrophic waters, others are parasitic or symbiotic on the gills, in the blood, or on the body surface of aquatic animals, causing damage to various physiological functions of the host (Song et al. 2003). In contrast, Peniculia is a relatively small subclass (about 100 species reported), with members of the genera Paramecium Müller, 1773 and Frontonia Ehrenberg, 1833 being the main contributors to peniculid species diversity. Peniculids usually have a larger body size (length over 50 μm) than scuticociliates, and the majority are free-living members either of the plankton or the benthos.

  • Results and remarks
  • In the present project, 13 species of scuticociliates and 11 species of peniculids have been isolated from LWW (Table 5). Most of these isolates are free-living, the exceptions being four scuticociliates (three as symbionts of molluscs and one as a facultative parasite of crabs). Six species were recognized as new species (unpublished, in preparation for publication), i.e., two species of Frontonia Ehrenberg, 1833, one species of Glauconema Thompson, 1966, one species of Marituja Gajewskaja, 1928, one species of Myxophyllum Raabe, 1934, and one species of Pleuronema Dujardin, 1841.

  • Species diversity of peritrichs (Figs. 8, 9; Table 6)

  • Figure 8.  Selected peritrich ciliates found in Lake Weishan Wetland. A, H, P Epistylis spp. B Ophrydium crassicaule. C Campanella umbellaria. D Protargol-stained zooid of Epistylis hentscheli. E, F Vorticella spp. G Epicarchesium sinense, arrows mark the discontinuous spasmoneme. I Campanella sinica. J Dry silver nitrate-stained zooid of Epicarchesium granulitum, arrow marks the trochal band. K, O Pyxicola carteri, arrow marks the operculum, arrowhead marks the tube in lorica base. L Opercularia nutans. M Part of oral ciliature of Carchesium polypinum (image processed by the invert function via Photoshop). N Zoothamnium arbuscula. Q Opercularia sp. Scale bars 200 μm (A, B), 150 μm (G), 80 μm (H, I), 30 μm (K, L), 40 μm (O), 70 μm (P), 30 μm (Q)

    Figure 9.  Selected parasitic/epibiotic peritrich ciliates found in Lake Weishan Wetland (original). A–D Epistylis spp., arrows indicate the zooids. E Trichodina pediculus (arrows) on Hydra sp. F Adhesive disk of Trichodina nobillis. G, K Carchesium cyclopidarum, protargol-stained specimen in G, arrows in K indicate colonies attached to the host. H Lagenophrys (?) sp. (arrows). I Adhesive disk of Trichodina sp. (after dry silver nitrate staining). J Trichodina sp. (arrows) on skin of a fish. Scale bars 0.5 cm in A; 100 μm (B); 60 μm (C, D); 20 μm (F, J)

  • Background
  • The subclass Peritrichia Stein, 1859 is probably the most speciose subclass in the class Oligohymenophorea, with more than 1000 nominal species that occupy a wide range of biotopes (Chen et al. 2022; Foissner et al. 1992; Kahl 1935; Wu et al. 2020, 2022b, c). Morphologically, peritrichs are mainly characterized by their conspicuous oral ciliature and vestigial somatic ciliature. The Peritrichia are divided into two orders: Sessilida and Mobilida Kahl, 1933. Most sessilids are sedentary and attach to a substrate via a stalk, a scopula, or a lorica, although some species are free-swimming. Sessilids may be free-living or epibiotic (Gómez et al. 2018; Lynn 2008; Mayén-Estrada and Dias 2021; Safi et al. 2022). Mobilids are characterized by the possession of an adhesive disk, a permanently ciliated trochal band, and all species are epibiotic (Wang et al. 2022b). Early molecular phylogenetic analyses of the Peritrichia based on SSU rDNA sequences indicated that the mobilids have a closer relationship with hymenostomatids than with sessilids (Gong et al. 2006; Zhan et al. 2009). However, this relationship was subsequently found to be an artifact (Zhan et al. 2013). Phylogenomic analyses subsequently provided strong support for the sister relationship between Sessilida and Mobilida and the monophyly of the subclass Peritrichia (Gentekaki et al. 2017; Jiang et al. 2019).

  • Results and remarks
  • In the present project, 58 peritrich species comprising 49 sessilids and nine mobilids have been found in the LWW, representing the most significant proportion of ciliate species recorded (Table 6). Several papers have been published that include descriptions of novel species, e.g., Campanella sinica Wang et al., 2021, Epistylis foissneri Wu et al., 2021, Epicarchesium sinense Wu et al., 2021, and Zoothamnium weishanicum Wu et al., 2021, or redescriptions of under-researched species (Wang et al. 2021, 2022b, c; Wu et al. 2021b, c). Several unidentified species, such as Orborhabdostyla sp. (in preparation for publication), are probably new. Two of these are unidentified species of the family Scyphidiidae which may represent new genera (in preparation for publication). Based on our newly obtained data, a few insufficiently known species will be redescribed or reclassified, including Epistylis daphniae Fauré-Fremiet, 1905, Epistylis lwoffi Fauré-Fremiet, 1943, Epistylis pygmaeum (Ehrenberg, 1838) Foissner et al., 1999, and Trichodina nigra Lom, 1960. Some epibiotic sessilids are morphologically similar in vivo to congeners that are free-living. Investigations of these species will provide new insights on the taxonomy and classification of sessilids.

  • Species diversity of hypotrichs (Fig. 10; Table 7)

  • Figure 10.  Selected hypotrichous ciliates found in Lake Weishan Wetland (original). A Cyrtohymena primicirrata. B Pleurotricha curdsi. C, J Stichotricha sp. D, E Kerona pediculus in vivo (D) and after protargol staining (E, image processed by the invert function via Photoshop). F Cortical granules of Pseudokeronopsis erythrina. G Euplotes muscicola. H Detail of silverline system on dorsal side of Euplotes sp. I Urosoma caudata. K, L Chaetospira sp. in vivo (K) and after protargol staining (L) Scale bars 50 μm (A, B, D, I–L), 20 μm (G)

  • Background
  • Hypotrichs are a large group, i.e., with about 200 genera and 1000 nominal species, of morphologically diverse and ubiquitously distributed ciliates that are widely regarded as the most complex and highly differentiated group within the Ciliophora (Abraham et al. 2021; Jung et al. 2021; Omar et al. 2021; Shao et al. 2020; Vďačný and Foissner 2021; Xu et al. 2020). Most hypotrichs are recognizable by their dorso-ventrally flattened body, conspicuous adoral zone of membranelles, and compound ventral ciliary organelles known as cirri (Berger 1999, 2006, 2011; Curds 1975; Kahl 1932; Small and Lynn 1985; Wang et al. 2021b, c). Since the 1970s, the classification of hypotrichs has undergone several major revisions. According to Shao et al. (2020), Hypotrichia Stein, 1859 includes at least six orders, i.e., Discocephalida Wicklow, 1982, Euplotida Small & Lynn, 1985, Kiitrichida Nozawa, 1941, Stichotrichida Fauré-Fremiet, 1961, Sporadotrichida Fauré-Fremiet, 1961, and Urostylida Jankowski, 1979. To date, hypotrichs remain one of the most confused groups in terms of their systematics, mainly due to a lack of detailed morphological, ontogenetic, and molecular data for many taxa (Berger 1999, 2006, 2011; Foissner and Berger 2021; Foissner et al. 1991; Paiva 2020; Shao et al. 2020; Zhang et al. 2020).

  • Results and remarks
  • In the present study, detailed taxonomic data of 37 hypotrichs (Table 7), including 32 species of hypotrichs s. str. and five euplotids, have been acquired, with morphogenetic information for ten species. Details of the morphology, ontogenesis, and molecular data of two novel species, i.e., Metaurostylopsis alrasheidi Song et al., 2020 and Heterobakuella bergeri Song et al., 2021, have been published (Song et al. 2020, 2021b). Another two species, Chaetospira sp. and Pseudouroleptus sp. 1, are also thought to be new species and a novel species of Deviatidae possibly represents a new genus. In addition, more than 15 of these investigated species have also been found in other countries or in different localities within China.

Conclusion, limitations, and prospects
  • Among the groups involved in the present project, Oligohymenophorea has the largest number of species followed by Spirotrichea. However, this distribution will likely need to be updated or refreshed because investigations of the LWW ciliates are ongoing. The molecular ecological results also imply that there is a higher species diversity than that revealed by morphological studies. Many species found in the LLW have previously been found in other continents, providing evidence for the cosmopolitan distribution model of ciliates (Finlay 2002; Finlay and Fenchel 2004). Additionally, the comprehensive and detailed taxonomic data obtained in the current project will facilitate accurate species-level identifications for future molecular ecological analyses.

    In addition to the taxa mentioned above, a considerable number of ciliates isolated from the LWW have not been well investigated due to a lack of time and expertise, and difficulties in the application of the silver stain. Additionally, our published work has implied some investigated genera/families are non-monophyletic, thus their classification will be revised based on detailed morphological data and molecular analyses. Future work on ciliates in the LWW will focus on: (1) the production of a ciliate atlas including both free-living and parasitic/symbiotic species; (2) the evolutionary relationships of ciliates; (3) revisions of the systematics and classification of selected taxa resulting from the taxonomic and molecular phylogenetic investigations; (4) the process of succession of dominance by different groups of ciliates.

Materials and methods

    Sample collection and morphological methods

  • Samples comprising water, sediment, mud and/or rotting plants were collected from lakes, rivers, canals, brooks, wetlands, ports, and aquaculture ponds in the LWW, Shandong Province, China (Fig. 1). The samples were transferred to the laboratory of Weishan Wetland Station (34° 38′ 34″ N, 117° 16′ 46″ E) in bottles or tanks and checked under a dissecting microscope. For most species, raw cultures were established in Petri dishes (9 cm diameter) containing in situ water and a nutritional resource, such as grains of rice, wheat or millet, and incubated at room temperature (about 25 ℃). Living specimens were investigated using bright field and differential interference contrast microscopy (Olympus BX53; Olympus Corporation, Tokyo, Japan) at magnifications of 40–1000×. Protargol staining, dry silver nitrate staining, wet silver nitrate staining and silver carbonate staining were performed following Wilbert (1975), Klein (1958), Corliss (1953), and Ma et al. (2003), respectively. Classification mainly follows Lynn (2008). The charts shown in Fig. 3 were made by GENESCLOUD (www.genescloud.cn/).

  • DNA extraction and PCR amplification, and sequencing

  • Ciliate cells were isolated and washed several times in double distilled water using glass micropipettes under a dissecting microscope. At least one cell was used for extracting the total genomic DNA using a DNeasy Blood & Tissue Kit (Qiagen, Hilden, Germany). The DNA polymerase, primers, and programs used in PCR amplification were according to Chi et al.(2020, 2021a, b) and Wang et al. (2021). The fragments were sequenced bidirectionally by the Tsingke Biological Technology Company (Qingdao, China). Sequences of some species have been deposited in the GenBank database (for accession numbers, see Tables 1, 2, 3, 4, 5, 6 and 7). All DNA samples have been coded and stored at the Laboratory of Protozoology at the Ocean University of China.

  • Environmental and molecular ecological analyses

  • Taking into consideration that our biodiversity/taxonomic survey was especially lacking data concerning lake planktonic ciliates, we decided to make a preliminary screening of planktonic ciliate biodiversity using high-throughput sequencing. Results were only aimed at providing preliminary data for further study, either involving traditional taxonomic approaches or high-throughput sequencing.

    From August 2020 to July 2021, 5 L lake water (from ~ 50 cm below the water surface) was collected from LWW on each sampling occasion for environmental DNA extraction. Dissolved oxygen concentration (DO), electrical conductivity (EC), pH and temperature were measured in situ with a multi-parameter water quality sonde (YSI, USA). The surface water samples were immediately transported to the laboratory and filtered through 0.22 μm pore-size PVDF membranes (Millipore, Ireland) using a vacuum pump until the filter was completely clogged. Then, filters were immediately stored at − 80 ℃ until DNA extraction. The filtrate was used to measure the nitrate (NO3) concentration by ion chromatography (Thermo Scientific, USA).

    Total DNA was extracted in triplicate using the DNeasy PowerWater kit (Qiagen, USA) following the manufacturer's recommendations. Triplicate DNA extractions of each sample were combined and mixed to form a pooled sample. The primers EK-565F (5′-GCAGTTAAAAAGCTCGTAGT-3′) and EK-1134R (5′-TTTAAGTTTCAGCCTTGCG-3′) were used to amplify the hypervariable V4 region of the 18S rDNA (Bower et al. 2004; Simon et al. 2015). The triplicate PCR reactions and cycling parameters of the 18S amplicon were according to Wang et al. (2022). PCR products from triplicate reactions per sample were purified, pooled, and then sequenced on the Illumina MiSeq platform (Majorbio, Shanghai, China).

    Paired-end reads were merged with FLASH (Magoč and Salzberg 2011), trimmed with Trimmomatic (Bolger et al. 2014), and quality-filtered by QIIME v1.8 (Caporaso et al. 2010). Chimeric sequences were identified and removed by UCHIME (Edgar et al. 2011). Sequences that occurred only once in the entire data set (singletons) were also excluded. Operational taxonomic units (OTUs) were clustered with 97% sequence similarity cutoff using UPARSE (Edgar 2013). Taxonomic classification of the OTUs was carried out by RDP Classifier algorithm against the PR2 database (Guillou et al. 2012). Prior to statistical analyses, samples were rarefied to an even sampling depth by random resampling (McMurdie and Holmes 2013). Statistical analyses were conducted using the "vegan" package and visualized by the "ggplot2" package in R v.3.6.3 (Oksanen et al. 2014; Wickham 2016). Temporal variations in the plankton community composition of Ciliophora at the rank of order were analyzed by the "circlize" package.

  • Supplementary Information

  • The online version contains supplementary material available at https://doi.org/10.1007/s42995-022-00154-x.

  • Acknowledgements

  • This work was supported by the Laoshan Laboratory (No. LSKJ202203201), the National Natural Science Foundation of China (project numbers: 32030015, 32000300, 31961123002), the China Postdoctoral Science Foundation (project number: 2021M703037, 2022M711922), and the Researchers Supporting Project (Project number RSP2022R10) of the King Saud University, Saudi Arabia. Our thanks are also given to the "Weishan Wetland Station" for the institutional support.

  • Author contributions

  • WBS and HGM conceived and guided the study. YC, BRL, GATZ, TL, ZW, TW, WYS, YFW and YJL conducted the research; YC wrote the content of heterotrichs; BRL wrote the content of haptorids and prostomateans; GATZ wrote the content of pleurostomatids; TL wrote the content of hymenostomatids; ZW, TW wrote the content of peritrichs; WYS wrote the content of hypotrichs; YFW wrote the content of ecology; KASA and AW reviewed and edited the manuscript. All authors read and approved the final version of the manuscript.

  • Data availability

  • Most datasets generated or analyzed during this study are included in this published article (and its supplementary material), and the rest, e.g., DNA, some gene sequences, and morphological details, are available from the corresponding author upon reasonable request.

Declarations

    Conflict of interest

  • The authors declare that they have no conflict of interest. Author Alan Warren and Weibo Song are members of the Editorial Board, but they were not involved in the journal's review of, or decision related to, this manuscript.

  • Animal and human rights statements

  • We declare that all applicable international, national, and or institutional guidelines for sampling, care, and experimental use of organisms for the study have been followed and all necessary approvals have been obtained.

Reference (125)

Catalog

/

DownLoad:  Full-Size Img  PowerPoint
Return
Return