{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T20:04:28Z","timestamp":1792008268472,"version":"4.4.0"},"reference-count":33,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T00:00:00Z","timestamp":1790121600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T00:00:00Z","timestamp":1790121600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32403069"],"award-info":[{"award-number":["32403069"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32573565"],"award-info":[{"award-number":["32573565"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005046","name":"Natural Science Foundation of Heilongjiang Province","doi-asserted-by":"publisher","award":["YQ2024C022"],"award-info":[{"award-number":["YQ2024C022"]}],"id":[{"id":"10.13039\/501100005046","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002858","name":"China Postdoctoral Science Foundation","doi-asserted-by":"publisher","award":["2023MD744174"],"award-info":[{"award-number":["2023MD744174"]}],"id":[{"id":"10.13039\/501100002858","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010009","name":"Heilongjiang Provincial Postdoctoral Science Foundation","doi-asserted-by":"publisher","award":["LBH\u2010Z23089"],"award-info":[{"award-number":["LBH\u2010Z23089"]}],"id":[{"id":"10.13039\/501100010009","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                    Koi sleepy disease (KSD), also called carp edema virus disease (CEVD), is a widespread viral infection affecting common carp (\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Cyprinus carpio<\/jats:italic>\n                    <\/jats:styled-content>\n                    ) and its ornamental varieties, leading to substantial economic losses in global aquaculture. The development of effective prophylactic and therapeutic measures against CEVD has been hampered by the lack of a robust system for in\u00a0vitro propagation of its causative agent, carp edema virus (CEV). To address this constraint, a method was established for the isolation and continuous culture of CEV using primary head kidney cells derived from Songpu mirror carp. A highly pathogenic CEV strain, L\u20103, isolated from an outbreak in Liaoning, China, was successfully propagated in\u00a0vitro through progressive adaptation during serial passage. Pronounced cytopathic effects (CPE) were observed over 17 consecutive passages, with CEV presence confirmed in each passage by nested PCR. Viral replication dynamics, assessed across 20 successive passages, demonstrated a progressive increase in viral copy numbers from very low levels at initial passage to peak at passage 18, confirming successful adaptation to primary head kidney cells. Challenge trials with the 20th\u2010passage virus induced clinical signs characteristic of KSD in carp. Moreover, examination of gill tissues from infected fish by transmission electron microscopy revealed viral particles with morphology typical of CEV. The present work reveals a novel approach for the propagation of CEV in\u00a0vitro. These findings are expected to provide a critical tool for advancing fundamental research on viral pathogenesis and supporting the systematic development and evaluation of vaccines against CEVD.\n                  <\/jats:p>","DOI":"10.1111\/jfd.70296","type":"journal-article","created":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T01:11:04Z","timestamp":1790212264000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Establishment of a Novel Primary Cell Culture Model for the Continuous Propagation of Carp Edema Virus In\u00a0Vitro"],"prefix":"10.1111","author":[{"given":"Yue","family":"Liu","sequence":"first","affiliation":[{"name":"College of Animal Science and Technology Northeast Agricultural University  Harbin People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zuwei","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology Northeast Agricultural University  Harbin People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sixuan","family":"Peng","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology Northeast Agricultural University  Harbin People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kim Jong","family":"Su","sequence":"additional","affiliation":[{"name":"Department of Aquaculture Wonsan University of Fisherise  Wonsan City Kangwon Province Democratic People's Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuanyu","family":"Ren","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology Northeast Agricultural University  Harbin People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaonan","family":"Lv","sequence":"additional","affiliation":[{"name":"Beijing Aquaculture Technology Extention Station  Beijing People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinshan","family":"Huang","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology Northeast Agricultural University  Harbin People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xu","family":"Lin","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology Northeast Agricultural University  Harbin People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5861-8108","authenticated-orcid":false,"given":"Wen","family":"Shi","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology Northeast Agricultural University  Harbin People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology Northeast Agricultural University  Harbin People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,23]]},"reference":[{"issue":"6","key":"e_1_2_11_2_1","doi-asserted-by":"crossref","DOI":"10.1111\/jfd.13934","article-title":"Potential to Grow Carp Oedema Virus (Genogroup I) in Monolayers of Carp\u2010Derived Primary Cells With Further Implication in Cell Analysis","volume":"47","author":"Abdelsalam E. E. E.","year":"2024","journal-title":"Journal of Fish Diseases"},{"key":"e_1_2_11_3_1","article-title":"Carp Edema Virus: Host Selection and Interaction, and Potential Factors Affecting Its Introduction to the Common Carp Population, Distribution, Survival","volume":"563","author":"Abdelsalam E. E. E.","year":"2023","journal-title":"Veterinary Research"},{"key":"e_1_2_11_4_1","doi-asserted-by":"crossref","first-page":"245","DOI":"10.3354\/dao02994","article-title":"Concentration of Carp Edema Virus (CEV) DNA in Koi Tissues Affected by Koi Sleepy Disease (KSD)","volume":"119","author":"Adamek M.","year":"2016","journal-title":"Diseases of Aquatic Organisms"},{"key":"e_1_2_11_5_1","doi-asserted-by":"crossref","DOI":"10.1016\/j.virol.2025.110671","article-title":"Development of an Effective Dna Vaccine Against Ayu Atypical Cellular Gill Disease Using Draft Genome Information of the Causative Agent, \n                     plecoglossus altivelis\n                   Poxvirus","volume":"612","author":"Baba S.","year":"2025","journal-title":"Virology"},{"key":"e_1_2_11_6_1","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/S0044-8486(02)00570-7","article-title":"Establishment of a Pluripotent Embryonic Cell Line From Sea Perch (\n                     Lateolabrax japonicus\n                  ) Embryos","volume":"218","author":"Chen S. 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H.","year":"1995","journal-title":"Journal of General Virology"},{"issue":"12","key":"e_1_2_11_8_1","doi-asserted-by":"crossref","first-page":"4137","DOI":"10.1128\/JCM.00827-14","article-title":"Identification of a Novel RNA Virus Lethal to Tilapia","volume":"52","author":"Eyngor M.","year":"2014","journal-title":"Journal of Clinical Microbiology"},{"key":"e_1_2_11_9_1","doi-asserted-by":"crossref","first-page":"1767","DOI":"10.1111\/jfd.13698","article-title":"The Influence of Viral Infection on Cell Line Characteristics: Lessons Learned From Working With New Cell Lines From Common Carp","volume":"45","author":"Felten M.","year":"2022","journal-title":"Journal of Fish Diseases"},{"key":"e_1_2_11_10_1","unstructured":"Haenen O. K.Way B.Gorgoglione et\u00a0al.2016. \u201cNovel Viral Infections Threatening Cyprinid Fish.\u201d36:11\u201323."},{"key":"e_1_2_11_11_1","doi-asserted-by":"crossref","DOI":"10.1111\/jfd.70027","article-title":"Development of a Sensitive Real\u2010Time PCR Method for Detection of Carp Oedema Virus","volume":"49","author":"He H.","year":"2025","journal-title":"Journal of Fish Diseases"},{"key":"e_1_2_11_12_1","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/j.phrp.2015.11.011","article-title":"Traditional and Modern Cell Culture in Virus Diagnosis","volume":"7","author":"Hematian A.","year":"2016","journal-title":"Osong Public Health and Research Perspectives"},{"key":"e_1_2_11_13_1","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1007\/s00705-024-06168-3","article-title":"Virus Susceptibility of a New Cell Line Derived From the Muscle of Koi (\n                     cyprinus carpio\n                   Koi)","volume":"169","author":"Jing H.","year":"2024","journal-title":"Archives of Virology"},{"key":"e_1_2_11_14_1","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1186\/s12917-015-0424-7","article-title":"Another Potential Carp Killer?: Carp Edema Virus Disease in Germany","volume":"11","author":"Jung\u2010Schroers V.","year":"2015","journal-title":"BMC Veterinary Research"},{"key":"e_1_2_11_15_1","article-title":"Prolonged Airway Explant Culture Enables Study of Health, Disease, and Viral Pathogenesis","volume":"11","author":"Lee\u2010Ferris R. E.","year":"2025","journal-title":"Science Advances"},{"key":"e_1_2_11_16_1","doi-asserted-by":"crossref","first-page":"1944","DOI":"10.1111\/jfb.13277","article-title":"Establishment and Characterization of a Cell Line From the Head Kidney of Golden Pompano \n                     Trachinotus ovatus\n                   and Its Application in Toxicology and Virus Susceptibility","volume":"90","author":"Li P.","year":"2017","journal-title":"Journal of Fish Biology"},{"issue":"3","key":"e_1_2_11_17_1","doi-asserted-by":"crossref","first-page":"177","DOI":"10.3354\/dao03296","article-title":"First Report of Carp Edema Virus in the Mortality of Wild Common Carp \n                     Cyprinus carpio\n                   in North America","volume":"131","author":"Lovy J.","year":"2018","journal-title":"Diseases of Aquatic Organisms"},{"key":"e_1_2_11_18_1","doi-asserted-by":"crossref","first-page":"e221","DOI":"10.1128\/MRA.00239-21","article-title":"Complete Genome Sequence of Carp Edema Virus Isolated From Koi Carp","volume":"10","author":"Mekata T.","year":"2021","journal-title":"Microbiology Resource Announcements"},{"key":"e_1_2_11_19_1","doi-asserted-by":"crossref","first-page":"197","DOI":"10.3354\/dao065197","article-title":"Histopathological and Electron Microscopy Studies on Sleepy Disease of Koi \n                     Cyprinus carpio\n                   Koi in Japan","volume":"65","author":"Miyazaki T.","year":"2005","journal-title":"Diseases of Aquatic Organisms"},{"key":"e_1_2_11_20_1","doi-asserted-by":"crossref","DOI":"10.1111\/jfd.70119","article-title":"Development of an Effective Dna Vaccine Against Viral Edema of Carp\/Koi Sleepy Disease Caused by Carp Edema Virus","volume":"49","author":"Ogawa R.","year":"2026","journal-title":"Journal of Fish Diseases"},{"key":"e_1_2_11_21_1","doi-asserted-by":"crossref","first-page":"167","DOI":"10.3147\/jsfp.21.167","article-title":"A Histopathological Study on Juvenile Colorcarp, \n                     cyprinus carpio\n                  , Showing Edema","volume":"21","author":"Ono S.","year":"1986","journal-title":"Fish Pathology"},{"key":"e_1_2_11_22_1","doi-asserted-by":"crossref","DOI":"10.1016\/j.aquaculture.2025.742151","article-title":"Identification of a Novel Strain of Carp Edema Virus Affecting Songpu Mirror Carp in Liaoning, China","volume":"599","author":"Shi W.","year":"2025","journal-title":"Aquaculture"},{"key":"e_1_2_11_23_1","doi-asserted-by":"crossref","DOI":"10.3390\/v9010019","article-title":"Novel Approach for Isolation and Identification of Porcine Epidemic Diarrhea Virus (PEDV) Strain NJ Using Porcine Intestinal Epithelial Cells","volume":"9","author":"Shi W.","year":"2017","journal-title":"Viruses"},{"issue":"3","key":"e_1_2_11_24_1","doi-asserted-by":"crossref","first-page":"755","DOI":"10.1638\/2017-0189.1","article-title":"Outbreak and Treatment of Carp Edema Virus in Koi (\n                     Cyprinus carpio\n                  ) From Northern California","volume":"49","author":"Stevens B. N.","year":"2018","journal-title":"Journal of Zoo and Wildlife Medicine"},{"key":"e_1_2_11_25_1","doi-asserted-by":"crossref","DOI":"10.1128\/spectrum.01164-24","article-title":"Comparison of Mutations in Human Parainfluenza Viruses During Passage in Primary Human Bronchial\/Tracheal Epithelial Air\u2010Liquid Interface Cultures and Cell Lines","volume":"12","author":"Sugimoto S.","year":"2024","journal-title":"Microbiol Spectr"},{"issue":"12","key":"e_1_2_11_26_1","doi-asserted-by":"crossref","first-page":"3392","DOI":"10.1099\/jgv.0.000649","article-title":"Emergence of Carp Edema Virus in Cultured Ornamental Koi Carp, \n                     Cyprinus carpio\n                   Koi, in India","volume":"97","author":"Swaminathan T. R.","year":"2016","journal-title":"Journal of General Virology"},{"key":"e_1_2_11_27_1","doi-asserted-by":"crossref","DOI":"10.1016\/j.fsi.2024.109519","article-title":"Expression Analysis and Antiviral Activity of Koi Carp (\n                     Cyprinus carpio\n                  ) Viperin Against Carp Edema Virus (CEV)","volume":"148","author":"Wang X. W.","year":"2024","journal-title":"Fish & Shellfish Immunology"},{"key":"e_1_2_11_28_1","doi-asserted-by":"crossref","first-page":"155","DOI":"10.3354\/dao03164","article-title":"Emergence of Carp Edema Virus (CEV) and Its Significance to European Common Carp and Koi \n                     Cyprinus carpio","volume":"126","author":"Way K.","year":"2017","journal-title":"Diseases of Aquatic Organisms"},{"key":"e_1_2_11_29_1","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.dci.2018.07.007","article-title":"Establishment of Fin Cell Lines and Their Use to Study the Immune Gene Expression in Cyprinid Fishes With Different Ploidy in Rhabdovirus Infection","volume":"88","author":"Xiao J.","year":"2018","journal-title":"Developmental and Comparative Immunology"},{"key":"e_1_2_11_30_1","doi-asserted-by":"crossref","first-page":"762","DOI":"10.1177\/10406387211005749","article-title":"An Efficient Peptide\u2010Based ELISA for Differentiating Fowl Adenovirus 4\u2010Infected Chickens From Vaccinated Chickens","volume":"33","author":"Xie S.","year":"2021","journal-title":"Journal of Veterinary Diagnostic Investigation"},{"key":"e_1_2_11_31_1","first-page":"100","article-title":"Immune Evasion Strategies of Seneca Valley Virus: Mechanisms of Host Innate Immune Suppression","volume":"3","author":"Xie S.","year":"2025","journal-title":"Agriculture Communications"},{"key":"e_1_2_11_32_1","doi-asserted-by":"crossref","DOI":"10.3390\/v15051216","article-title":"Identification of Host Proteins Interacting With Ibv s1 Based on Tracheal Organ Culture","volume":"15","author":"Zhang H.","year":"2023","journal-title":"Viruses"},{"key":"e_1_2_11_33_1","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1016\/j.aquaculture.2023.739869","article-title":"Carp Edema Virus, a Great Threat for Carp and Koi Farming in Henan, China","volume":"576","author":"Zhang J.","year":"2023","journal-title":"Aquaculture"},{"key":"e_1_2_11_34_1","doi-asserted-by":"crossref","DOI":"10.1016\/j.aquaculture.2022.739007","article-title":"Routine Development of Long\u2010Term Primary Cell Culture and Finite Cell Line From the Hemolymph of Greasyback Shrimp (\n                     Metapenaeus ensis\n                  ) and Virus Susceptibility","volume":"563","author":"Zhao Y.","year":"2023","journal-title":"Aquaculture"}],"container-title":["Journal of Fish Diseases"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/jfd.70296","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/jfd.70296","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/jfd.70296","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T01:11:10Z","timestamp":1790212270000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/jfd.70296"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,23]]},"references-count":33,"alternative-id":["10.1111\/jfd.70296"],"URL":"https:\/\/doi.org\/10.1111\/jfd.70296","archive":["Portico"],"relation":{},"ISSN":["0140-7775","1365-2761"],"issn-type":[{"value":"0140-7775","type":"print"},{"value":"1365-2761","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,23]]},"assertion":[{"value":"2026-08-19","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-11","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-09-23","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70296"}}