{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T02:13:38Z","timestamp":1783649618163,"version":"3.55.0"},"reference-count":25,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T00:00:00Z","timestamp":1783555200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T00:00:00Z","timestamp":1783555200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100014188","name":"Ministry of Science and ICT, South Korea","doi-asserted-by":"publisher","award":["RS\u20102024\u201000405986"],"award-info":[{"award-number":["RS\u20102024\u201000405986"]}],"id":[{"id":"10.13039\/501100014188","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Fish Diseases"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Scuticociliatosis caused by the ciliate\n                    <jats:italic>Miamiensis avidus<\/jats:italic>\n                    remains a serious constraint in the intensive culture of olive flounder (\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Paralichthys olivaceus<\/jats:italic>\n                    <\/jats:styled-content>\n                    ), yet effective vaccines are still unavailable. In this study, we evaluated chemical attenuation as a novel approach to generate a safe whole\u2010cell vaccine candidate against\n                    <jats:italic>M. avidus<\/jats:italic>\n                    . Ciliates were attenuated by oxytetracycline (OTC) treatment, and the safety, residual virulence and protective potential of the chemically attenuated preparation were assessed in\u00a0vitro and in\u00a0vivo. Treatment with 5\u2009mM OTC for 3\u2009h suppressed ciliate proliferation while preserving motility and morphology. The attenuated ciliates exhibited markedly reduced cytopathic effects on EPC feeder cells and caused no mortality in olive flounder, confirming successful attenuation. Fish immunised with the chemically attenuated whole\u2010cell vaccine candidate of\n                    <jats:italic>M. avidus<\/jats:italic>\n                    showed reduced cumulative mortality following experimental challenge compared with PBS\u2010injected controls and fish immunised with non\u2010adjuvanted formalin\u2010killed cells, particularly when a booster immunisation was administered. Serum agglutination titre was significantly different between the PBS control and vaccinated groups, but not among the vaccinated groups. Overall, these results suggest, for the first time, that chemical attenuation can be applied to a scuticociliate parasite to generate a safe whole\u2010cell vaccine candidate that provides partial protection. This proof\u2010of\u2010concept study highlights the potential of chemical attenuation as a foundation for further optimisation of vaccine strategies against scuticociliatosis.\n                  <\/jats:p>","DOI":"10.1111\/jfd.70242","type":"journal-article","created":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T01:27:21Z","timestamp":1783646841000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Chemical Attenuation of\n                    <i>Miamiensis avidus<\/i>\n                    Using Oxytetracycline Reduces Virulence and Confers Partial Protection in Olive Flounder (\n                    <i>Paralichthys olivaceus<\/i>\n                    )"],"prefix":"10.1111","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5587-2496","authenticated-orcid":false,"given":"Eun Gyeong","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Aquatic Life Medicine Pukyong National University  Busan South Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1140-8162","authenticated-orcid":false,"given":"Ki Hong","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Aquatic Life Medicine Pukyong National University  Busan South Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,7,9]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"crossref","first-page":"177","DOI":"10.14411\/fp.2004.021","article-title":"Histophagous Scuticociliatids (Ciliophora) Parasitizing Turbot \n                     Scophthalmus maximus\n                  : Morphology, In\u00a0Vitro Culture and Virulence","volume":"51","author":"Alvarez\u2010Pellitero P.","year":"2004","journal-title":"Folia Parasitologica"},{"key":"e_1_2_11_3_1","doi-asserted-by":"crossref","DOI":"10.1038\/s41541-025-01165-2","article-title":"Live Attenuated SARS\u2010CoV\u20102 Vaccine OTS\u2010228 Demonstrates Efficacy, Safety and Stability in a Preclinical Model","volume":"10","author":"Britzke T.","year":"2025","journal-title":"npj Vaccines"},{"key":"e_1_2_11_4_1","doi-asserted-by":"crossref","DOI":"10.1038\/ncomms15480","article-title":"Design of Live Attenuated Bacterial Vaccines Based on D\u2010Glutamate Auxotrophy","volume":"8","author":"Cabral M. P.","year":"2017","journal-title":"Nature Communications"},{"key":"e_1_2_11_5_1","doi-asserted-by":"crossref","first-page":"6891","DOI":"10.1128\/AAC.01779-15","article-title":"Novel Insights Into the Molecular Events Linking to Cell Death Induced by Tetracycline in the Amitochondriate Protozoan Trichomonas vaginalis","volume":"59","author":"Huang K. Y.","year":"2015","journal-title":"Antimicrobial Agents and Chemotherapy"},{"key":"e_1_2_11_6_1","doi-asserted-by":"crossref","first-page":"47","DOI":"10.3354\/dao046047","article-title":"Philasterides dicentrarchi (Ciliophora, Scuticociliatida) as the Causative Agent of Scuticociliatosis in Farmed Turbot \n                     Scophthalmus maximus\n                   in Galicia (NW Spain)","volume":"46","author":"Iglesias R.","year":"2001","journal-title":"Diseases of Aquatic Organisms"},{"key":"e_1_2_11_7_1","doi-asserted-by":"crossref","first-page":"191","DOI":"10.3354\/dao049191","article-title":"Antiprotozoals Effective In\u00a0Vitro Against the Scuticociliate Fish Pathogen Philasterides dicentrarchi","volume":"49","author":"Iglesias R.","year":"2002","journal-title":"Diseases of Aquatic Organisms"},{"key":"e_1_2_11_8_1","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1017\/S0031182002002688","article-title":"Philasterides dicentrarchi (Ciliophora: Scuticociliatida) Expresses Surface Immobilization Antigens That Probably Induce Protective Immune Responses in Turbot","volume":"126","author":"Iglesias R.","year":"2003","journal-title":"Parasitology"},{"key":"e_1_2_11_9_1","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1016\/j.aquaculture.2013.09.017","article-title":"Evaluation of Treatment Efficacy of Doxycycline and Albendazole Against Scuticociliatosis in Olive Flounder \n                     Paralichthys olivaceus","volume":"416","author":"Kang Y. J.","year":"2013","journal-title":"Aquaculture"},{"key":"e_1_2_11_10_1","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.aquaculture.2008.03.015","article-title":"Optimization of an Inactivated Vaccine Against a Scuticociliate Parasite of Turbot: Effect of Antigen, Formalin and Adjuvant Concentration on Antibody Response and Protection Against the Pathogen","volume":"278","author":"Lamas J.","year":"2008","journal-title":"Aquaculture"},{"key":"e_1_2_11_11_1","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/0165-2427(95)05445-C","article-title":"Efficacy of a Live Cryptobia salmositica Vaccine and the Mechanism of Protection in Vaccinated Rainbow Trout \n                     Oncorhynchus mykiss\n                   Against Cryptobiosis","volume":"48","author":"Li S.","year":"1995","journal-title":"Veterinary Immunology and Immunopathology"},{"key":"e_1_2_11_12_1","doi-asserted-by":"crossref","DOI":"10.1128\/IAI.00587-18","article-title":"Controlled Infection Immunization Using Delayed Death Drug Treatment Elicits Protective Immune Responses to Blood\u2010Stage Malaria Parasites","volume":"87","author":"Low L. M.","year":"2018","journal-title":"Infection and Immunity"},{"key":"e_1_2_11_13_1","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1016\/j.parint.2017.10.007","article-title":"Analysis of Genes Encoding High\u2010Antigenicity Polypeptides in Three Serotypes of Miamiensis avidus","volume":"67","author":"Motokawa S.","year":"2018","journal-title":"Parasitology International"},{"key":"e_1_2_11_14_1","doi-asserted-by":"crossref","first-page":"1557","DOI":"10.1292\/jvms.10-0214","article-title":"Experimental Scuticociliatosis in Japanese Flounder \n                     Paralichthys olivaceus\n                   Infected With Miamiensis avidus: Pathological Study on the Possible Neural Routes of Invasion and Dissemination of the Scuticociliate Inside the Fish Body","volume":"72","author":"Moustafa E. M.","year":"2010","journal-title":"Journal of Veterinary Medical Science"},{"key":"e_1_2_11_15_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3147\/jsfp.53.1","article-title":"Extracellular Proteinases of Miamiensis avidus Causing Scuticociliatosis Are Potential Virulence Factors","volume":"53","author":"Narasaki Y.","year":"2018","journal-title":"Fish Pathology"},{"key":"e_1_2_11_16_1","doi-asserted-by":"crossref","first-page":"123","DOI":"10.3354\/dao02102","article-title":"Antibody Responses of Turbot \n                     Psetta maxima\n                   Against Various Antigen Formulations of Scuticociliates (Ciliophora)","volume":"86","author":"Palenzuela O.","year":"2009","journal-title":"Diseases of Aquatic Organisms"},{"key":"e_1_2_11_17_1","doi-asserted-by":"crossref","first-page":"4151","DOI":"10.1038\/s41598-021-83704-0","article-title":"Broad Cross Protection by Recombinant Live Attenuated Influenza H3N2 Seasonal Virus Expressing Conserved M2 Extracellular Domain in a Chimeric Hemagglutinin","volume":"11","author":"Park B. R.","year":"2021","journal-title":"Scientific Reports"},{"key":"e_1_2_11_18_1","doi-asserted-by":"crossref","DOI":"10.1128\/IAI.00062-17","article-title":"Chemical Attenuation in the Development of a Whole\u2010Organism Malaria Vaccine","volume":"85","author":"Raja A. I.","year":"2017","journal-title":"Infection and Immunity"},{"key":"e_1_2_11_19_1","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1111\/j.1365-2761.2007.00876.x","article-title":"Vaccination of Turbot \n                     Psetta maxima\n                   (L.) Against the Protozoan Parasite Philasterides dicentrarchi: Effects on Antibody Production and Protection","volume":"31","author":"Sanmart\u00edn M. L.","year":"2008","journal-title":"Journal of Fish Diseases"},{"key":"e_1_2_11_20_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.fsi.2007.06.007","article-title":"Immune Response of Turbot \n                     Psetta maxima\n                   (L.) (Pisces: Teleostei) to Formalin\u2010Killed Scuticociliates (Ciliophora) and Adjuvanted Formulations","volume":"24","author":"Sitj\u00e0\u2010Bobadilla A.","year":"2008","journal-title":"Fish & Shellfish Immunology"},{"key":"e_1_2_11_21_1","doi-asserted-by":"crossref","first-page":"1027","DOI":"10.1111\/j.1365-2761.2009.01089.x","article-title":"Antigenic Differences of the Scuticociliate Miamiensis avidus From Japan","volume":"32","author":"Song J. Y.","year":"2009","journal-title":"Journal of Fish Diseases"},{"key":"e_1_2_11_22_1","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1017\/S0031182000060066","article-title":"The Effects of Oxytetracycline on Theileria parva In\u00a0Vitro","volume":"100","author":"Spooner P. R.","year":"1990","journal-title":"Parasitology"},{"key":"e_1_2_11_23_1","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1017\/S0031182000060583","article-title":"Oxytetracycline Inhibition of Mitochondrial Protein Synthesis in Bovine Lymphocytes Infected With Theileria parva or Stimulated by Mitogen","volume":"101","author":"Spooner P. R.","year":"1990","journal-title":"Parasitology"},{"key":"e_1_2_11_24_1","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1186\/s12916-018-1173-9","article-title":"Vaccination With Chemically Attenuated Plasmodium falciparum Asexual Blood\u2010Stage Parasites Induces Parasite\u2010Specific Cellular Immune Responses in Malaria\u2010Na\u00efve Volunteers: A Pilot Study","volume":"16","author":"Stanisic D. I.","year":"2018","journal-title":"BMC Medicine"},{"key":"e_1_2_11_25_1","doi-asserted-by":"crossref","first-page":"87","DOI":"10.3354\/dao03678","article-title":"Scuticociliatosis Caused by Philasterides dicentrarchi","volume":"150","author":"Steverding D.","year":"2022","journal-title":"Diseases of Aquatic Organisms"},{"key":"e_1_2_11_26_1","doi-asserted-by":"crossref","first-page":"752","DOI":"10.2307\/3283000","article-title":"In Vitro Attenuation of Cryptobia salmositica and Its Use as a Live Vaccine Against Cryptobiosis in \n                     Oncorhynchus mykiss","volume":"76","author":"Woo P. T.","year":"1990","journal-title":"Journal of Parasitology"}],"container-title":["Journal of Fish Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/jfd.70242","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/jfd.70242","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/jfd.70242","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T01:27:24Z","timestamp":1783646844000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/jfd.70242"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,9]]},"references-count":25,"alternative-id":["10.1111\/jfd.70242"],"URL":"https:\/\/doi.org\/10.1111\/jfd.70242","archive":["Portico"],"relation":{},"ISSN":["0140-7775","1365-2761"],"issn-type":[{"value":"0140-7775","type":"print"},{"value":"1365-2761","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,9]]},"assertion":[{"value":"2026-01-08","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-07-03","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-07-09","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70242"}}