Abstract
Tetrahymenosis is caused by the ciliated protozoan Tetrahymena and is responsible for serious economic losses to the aquaculture industry worldwide. However, information regarding the molecular mechanism leading to tetrahymenosis is limited. In previous transcriptome sequencing work, it was found that one of the two β-tubulin genes in T. pyriformis was significantly expressed in infected fish, we speculated that β-tubulin is involved in T. pyriformis infecting fish. Herein, the potential biological function of the β-tubulin gene in Tetrahymena species when establishing infection in guppies was investigated by cloning the full-length cDNA of this T. pyriformis β-tubulin (BTU1) gene. The full-length cDNA of T. pyriformis BTU1 gene was 1873 bp, and the ORF occupied 1134 bp, whereas 5′ UTR 434 bp, and 3′ UTR 305 bp whose poly (A) tail contained 12 bases. The predicted protein encoded by T. pyriformis BTU1 gene had a calculated molecular weight of 42.26 kDa and pI of 4.48. Moreover, secondary structure analysis and tertiary structure prediction of BTU1 protein were also conducted. In addition, morphology, infraciliature, phylogeny, and histopathology of T. pyriformis isolated from guppies from a fish market in Harbin were also investigated. Furthermore, qRT-PCR analysis and experimental infection assays indicated that the expression of BTU1 gene resulted in efficient cell proliferation during infection. Collectively, our data revealed that BTU1 is a key gene involved in T. pyriformis infection in guppies, and the findings discussed herein provide valuable insights for future studies on tetrahymenosis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Astrofsky KM, Schech JM, Sheppard BJ, Obenschain CA, Chin AM, Kacergis MC, Laver ER, Bartholomew JL, Fox JG (2002) High mortality due to Tetrahymena sp. infection in laboratory-maintained zebrafish (Brachydanio rerio). Comparative Med 52:363–367. https://doi.org/10.1053/svms.2002.34642
Barahona I, Galego L, Jacquet M, Rodrigues-Pousada C (1985) Cloning of two β-tubulin related genes in Tetrahymena pyriformis. FEBS Lett 188:227–232. https://doi.org/10.1016/0014-5793(85)80377-X
Chantangsi C, Lynn DH (2008) Phylogenetic relationships within the genus Tetrahymena inferred from the cytochrome c oxidase subunit 1 and the small subunit ribosomal RNA genes. Mol Phylogenet Evol 49:979–987. https://doi.org/10.1016/j.ympev.2008.09.017
Chettri JK, Leibowitz MP, Ofir R, Zilberg D (2009) Protective immunization against Tetrahymena sp. infection in guppies (Poecilia reticulata). Fish Shellfish Immun 27(2):302–308. https://doi.org/10.1016/j.fsi.2009.05.013
Cóias R, Galego L, Barahona I, Rodrigues-Pousada C (1988) Destabilization of tubulin mRNA during heat shock in Tetrahymena pyriformis. Eur J Biochem 175:467–474. https://doi.org/10.1111/j.1432-1033.1988.tb14218.x
Corliss J (1973) History, taxonomy, ecology, and evolution of species of Tetrahymena. Biology of Tetrahymena
Coulson RM, Connor V, Chen JC, Ajioka JW (1996) Differential expression of Leishmania major β-tubulin genes during the acquisition of promastigote infectivity. Molecular and Biochemical Parasitology Mol Biochem Parasit 82:227–236. https://doi.org/10.1016/0166-6851(96)02739-9
Dobrzańska J (1959) The occurrence of ciliates of the genus Tetrahymena Furgason, 1940 in fresh-water Mussels. Bulletin de l Académie Polonaise des Sciences 66:377–382
Doerder FP (2019) Barcodes reveal 48 new species of Tetrahymena, Dexiostoma, and Glaucoma: phylogeny, ecology, and biogeography of new and established species. J Eukaryot Microbiol 66:182–208. https://doi.org/10.1111/jeu.12642
Dupuis P (1992) The beta-tubulin genes of Paramecium are interrupted by two 27 bp introns. The EMBO Journal 11(10):3713–3719
Edgerton B, O'Donoghue P, Wingfield M, Owens L (1996) Systemic infection of freshwater crayfish Cherax quadricarinatus by hymenostome ciliates of the Tetrahymena pyriformis complex. Dis Aquat Organ 27:123–129. https://www.int-res.com/articles/dao/27/d027p123.pdf
Ferguson HW, Hicks BD, Lynn DH, Ostland VE, Bailey J (1987). Cranial ulceration in Atlantic salmon Salmon salar associated with Tetrahymena sp. Dis Aquat Organ 2:191–195. https://www.int-res.com/articles/dao/2/d002p191.pdf
Foissner W, Berger H, Kohmann F (1994) Taxonomische und ökologische Revision der Ciliaten des Saprobiensystems. – Band III: Hymenostomata, Prostomatida, Nassulida. Informationsberichte des Bayerischen Landesamtes für Wasserwirtschaft, Heft 1/94 p 1–548. https://doi.org/10.1016/S0932-4739(11)80223-3
Foissner W (1992). The silver carbonate methods. Protocols in Protozoology Society of Protozoologists, Allen Press Inc pp C7.1–7.3.
Fong D, Wallach M, Keithly J, Melera PW, Chang KP (1984) Differential expression of mRNAs for alpha-and beta-tubulin during differentiation of the parasitic protozoan Leishmania mexicana. P Natl A Sci 81:5782–5786. https://doi.org/10.1073/pnas.81.18.5782
Fuchs MA, Ryan LA, Chambers EL, Moore CM, Fairweather I, Trudgett A, Timson DJ, Timson G, Hoey EM (2013) Differential expression of liver fluke β-tubulin isotypes at selected life cycle stages. Int J Parasitol 43:1133–1139. https://doi.org/10.1016/j.ijpara.2013.08.007
Gaertig J (2000) Molecular mechanisms of microtubular organelle assembly in Tetrahymena. J Eukaryot Microbiol 47:185–190. https://doi.org/10.1111/j.1550-7408.2000.tb00037.x
Goven BA, Dawe DL, Gratzek JJ (1981) Protection of channel catfish, Ictalurus punctatus against Ichthyophthirius multifiliis Fouquet by immunisation with varying doses of Tetrahymena pyriformis (Lwoff) cilia. Aquaculture 23:269–273. https://doi.org/10.1016/0044-8486(81)90020-X
Han JE, Tang KF, Kim JH (2018) The use of beta-tubulin gene for phylogenetic analysis of the microsporidian parasite Enterocytozoon hepatopenaei (EHP) and in the development of a nested PCR as its diagnostic tool. Aquaculture 495:899–902. https://doi.org/10.1016/j.aquaculture.2018.06.059
Harikrishnan R, Balasundaram C, Heo MS (2010) Scuticociliatosis and its recent prophylactic measures in aquaculture with special reference to South Korea: taxonomy, diversity and diagnosis of scuticociliatosis: part I control strategies of scuticociliatosis: Part II. Fish Shellfish Immun 29:15–31. https://doi.org/10.1016/j.fsi.2010.02.026
Herbert B, Graham P (2008) Tetrahymenosis, columnaris disease and motile aeromonad septicaemia in golden perch, Macquaria ambiqua (Richardson), from Australia. Diseases in Asian aquaculture VI 179–192. https://www.fhs-afs.net/daa_vi_files/13.pdf
Janke C, Magiera MM (2020) The tubulin code and its role in controlling microtubule properties and functions. Nat Rev Mol Cell Bio 21:307–326. https://doi.org/10.1038/s41580-020-0214-3
Knossow M, Campanacci V, Khodja LA, Gigant B (2020) The mechanism of tubulin assembly into microtubules: insights from structural studies. Iscience 23:101511. https://doi.org/10.1016/j.isci.2020.101511
Kurup SP, Tewari AK (2012) Induction of protective immune response in mice by a DNA vaccine encoding Trypanosoma evansi beta tubulin gene. Vet Parasitol 187:9–16. https://doi.org/10.1016/j.vetpar.2012.01.009
Lawhavinit O, Chukanhom K, Hatai K (2002) Effect of Tetrahymena on the occurrence of achlyosis in the guppy Poecilia reticulata. Mycoscience 43:27–31. https://doi.org/10.1007/s102670200005
Leibowitz MP, Zilberg D (2009) Tetrahymena sp. infection in guppies, Poecilia reticulata Peters: parasite characterization and pathology of infected fish. J Fish Dis 32:845–855. https://doi.org/10.1111/j.1365-2761.2009.01062.x
Leibowitz MP, Ofir R, Golan-Goldhirsh A, Zilberg D (2009) Cysteine proteases and acid phosphatases contribute to Tetrahymena spp. pathogenicity in guppies. Poecilia reticulata. Vet Parasitol 166:21–26. https://doi.org/10.1016/j.vetpar.2009.08.005
Lewis SA, Lee MGS, Cowan NJ (1985) Five mouse tubulin isotypes and their regulated expression during development. J Cell Biol 101:852–861. https://doi.org/10.1083/JCB.101.3.852
Li SQ, Fung MC, Reid SA, Inoue N, Lun ZR (2007) Immunization with recombinant beta-tubulin from Trypanosoma evansi induced protection against T. evansi, T. equiperdum and T. b. brucei infection in mice. Parasite Immunol 29:191–199. https://doi.org/10.1111/j.1365-3024.2006.00933.x
Li S, Hong SJ, Choi MH, Hong ST (2009) Identification of two β-tubulin isotypes of Clonorchis sinensis. Parasitol Res 105:1015–1021. https://doi.org/10.1007/s00436-009-1512-z
Li W, Warren A, Zhang S, Pan M, Xiao X, Dong Z, Liang J, Pan X (2021) Identification and infection mechanism of Tetrahymena vorax affecting the goldfish Carassius auratus. Aquaculture 539:736643. https://doi.org/10.1016/j.aquaculture.2021.736643
Liu M, Fan X, Gao F, Gao S, Yu Y, Warren A, Huang J (2016) Tetrahymena australis (Protozoa, Ciliophora): a well-known but “non-existing” taxon, consideration on its identification, definition and systematic position. J Eukaryot Microbiol 63:760–770. https://doi.org/10.1111/jeu.12323
Lynn DH, Doerder FP (2012) The life and times of Tetrahymena. Method Cell Biol 109:9–27. https://doi.org/10.1016/B978-0-12-385967-9.00002-5
Lynn D, Doerder F, Gillis P, Prosser R (2018) Tetrahymena glochidiophila n. sp., a new species of Tetrahymena (Ciliophora) that causes mortality to glochidia larvae of freshwater mussels (Bivalvia). Dis Aquat Organ 127:125–136. https://doi.org/10.3354/dao03188
Magiera MM, Singh P, Janke C (2018) SnapShot: functions of tubulin posttranslational modifications. Cell 173:1552–1552. https://doi.org/10.1016/j.cell.2018.05.032
Minoura I, You H, Yamakita Y, Takazaki H, Ayukawa R, Uchimura S, Muto E (2013) Overexpression, purification, and functional analysis of recombinant human tubulin dimer. FEBS Lett 587:3450–3455. https://doi.org/10.1016/j.febslet.2013.08.032
Msiska Z, Morton JB (2009) Isolation and sequence analysis of a β-tubulin gene from arbuscular mycorrhizal fungi. Mycorrhiza 19:501–513. https://doi.org/10.1007/s00572-009-0248-z
Nylander JAA (2004) MrModeltest v2. Distributed by the Author. Department of Systematic Zoology, Evolutionary Biology Centre, Uppsala University
Pan X, Li W, Warren A, Dong Z (2022) Tetrahymena pyriformis infection in the tiger barb, Puntius tetrazona: parasite characterization and pathology of infected fish. Aquaculture 549:737725. https://doi.org/10.1016/j.aquaculture.2021.737725
Pimenta LM, Ariav R, Zilberg D (2005) Environmental and physiological conditions affecting Tetrahymena sp. infection in guppies. Poecilia reticulata Peters. J Fish Dis 28:539–547. https://doi.org/10.1111/j.1365-2761.2005.00658.x
Ponpornpisit A, Endo M, Murata H (2000) Experimental infections of a ciliate Tetrahymena pyriformis on ornamental fishes. Fisheries Sci 66:1026–1031. https://www.jstage.jst.go.jp/article/fishsci1994/66/6/66_6_1026/_pdf
Pucciarelli S, Ballarini P, Sparvoli D, Barchetta S, Yu T, Detrich HW, Miceli C (2012) Distinct functional roles of β-tubulin isotypes in microtubule arrays of Tetrahymena thermophila, a model single-celled organism. Plos One 7:e39694. https://doi.org/10.1371/journal.pone.0039694
Rataj M, Van P (2020) Multi-gene phylogeny of Tetrahymena refreshed with three new histophagous species invading freshwater planarians. Parasitol Res 119:1523–1545. https://doi.org/10.1007/s00436-020-06628-0
Ronquist F, Teslenko M, Van Der Mark P, Ayres DL, Darling A, Höhna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP (2012) MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Syst Biol 61:539–542. https://doi.org/10.1093/sysbio/sys029
Schwenkenbecher JM, Albonico M, Bickle Q, Kaplan RM (2007) Characterization of beta-tubulin genes in hookworms and investigation of resistance-associated mutations using real-time PCR. Mol Biochem Parasit 156:167–174. https://doi.org/10.1016/j.molbiopara.2007.07.019
Sharon G, Leibowitz MP, Chettri JK, Isakov N, Zilberg D (2014) Comparative study of infection with Tetrahymena of different ornamental fish species. J Comp Pathol 150:316–324. https://doi.org/10.1016/j.jcpa.2013.08.005
Smith JJ, Yakisich JS, Kapler GM, Cole ES, Romero DP (2004) A β-tubulin mutation selectively uncouples nuclear division and cytokinesis in Tetrahymena thermophila. Eukaryot Cell 3:1217–1226. https://doi.org/10.1128/ec.3.5.1217-1226.2004
Soares H, Cyrne L, Barahona I, Rodrigues-Pousada C (1991) Different patterns of expression of β-tubulin genes in Tetrahymena pyriformis during reciliation. Eur J Biochem 197:291–299. https://doi.org/10.1111/j.1432-1033.1991.tb15910.x
Soares H, Galego L, Cóias R, Rodrigues-Pousada C (1993) The mechanisms of tubulin messenger regulation during Tetrahymena pyriformis reciliation. J Biol Chem 268:16623–16630. https://doi.org/10.1016/S0021-9258(19)85464-9
Stamatakis A, Hoover P, Rougemont J (2008) A rapid bootstrap algorithm for the RAxML web servers. Syst Biol 57:758–771. https://doi.org/10.1080/10635150802429642
Thazhath R, Liu C, Gaertig J (2002) Polyglycylation domain of β-tubulin maintains axonemal architecture and affects cytokinesis in Tetrahymena. Nat Cell Biol 4:256–259. https://doi.org/10.1038/ncb764
Thilakaratne ID, Rajapaksha G, Hewakopara A, Rajapakse RP, Faizal AC (2003) Parasitic infections in freshwater ornamental fish in Sri Lanka. Dis Aquat Organ 31:157–62. https://www.int-res.com/articles/dao2003/54/d054p157.pdf
Wang Z, Dong Z, Zhang D, Liang L, Mu W (2021) Histopathological parameters, antioxidant enzyme levels, transcriptome, and hematology parameters of Amur minnow (Phoxinus lagowskii) infection with Tetrahymena pyriformis. Aquacult Int 29:2635–2659. https://doi.org/10.1007/s10499-021-00772-9
Wilbert N (1975) Eine verbesserte technik der protargolimpr¨agnation für ciliaten. Mikrokosmos 64:171–179. http://refhub.elsevier.com/S0044-8486(21)00305-7/rf0910
Xia L, Hai B, Gao Y, Burnette D, Thazhath R, Duan J, Brḗ MH, Levilliers N, Gorovsky MA, Gaertig J (2000) Polyglycylation of tubulin is essential and affects cell motility and division in Tetrahymena thermophila. J Cell Biol 149:1097–1106. https://doi.org/10.1083/jcb.149.5.1097
Ye Z, Ni W, Zhang J, Zhang Y, Yu L, Huang X (2022) Molecular characterization of a profilin gene from a parasitic ciliate Cryptocaryon irritans. Exp Parasitol 236:108248. https://doi.org/10.1016/j.exppara.2022.108248
Zhan Z, Stoeck T, Dunthorn M, Xu K (2014) Identification of the pathogenic ciliate Pseudocohnilembus persalinus (Oligohymenophorea: Scuticociliatia) by fluorescence in situ hybridization. Eur J Protistol 50:16–24. https://doi.org/10.1016/j.ejop.2013.09.004
Acknowledgements
Many thanks are given to the student of Harbin Normal University, Wenyu Li, for her help on details of experiment. We would like to thank TopEdit (www.topeditsci.com) for English language editing of this manuscript.
Funding
This work was supported by the Natural Science Foundation of China (project numbers: 31970498).
Author information
Authors and Affiliations
Contributions
MJ: conceptualization, methodology, and writing—original draft preparation; CZ, SW, LL, and SZ: data curation, visualization, and investigation. LW: supervision. XP: writing—reviewing and editing.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Declarations
Ethical approval
The animal experiments were conducted following the Guide for the Care and Use of Laboratory Animals, the protocol of which was approved by the Harbin Normal University.
Consent to participate
All authors consent to participate in this publication.
Consent for publication
All authors consent to publish the manuscript.
Conflict of interest
The authors declare no competing interests.
Additional information
Handling Editor: Una Ryan.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
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.
About this article
Cite this article
Jiang, M., Zhou, C., Wang, S. et al. Identification of a Tetrahymena species infecting guppies, pathology, and expression of beta-tubulin during infection. Parasitol Res 123, 104 (2024). https://doi.org/10.1007/s00436-024-08117-0
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s00436-024-08117-0