{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T05:52:06Z","timestamp":1785909126783,"version":"3.56.0"},"reference-count":29,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:00:00Z","timestamp":1785888000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p>Covert mortality nodavirus (CMNV) is the pathogen causing viral covert mortality disease (VCMD) in cultured shrimp, and cross-species transmission is an important characteristic of this virus. The lack of stable, continuously passable shrimp cell lines has long been a bottleneck for in-depth studies on CMNV pathogenesis, virus-host interactions, and antiviral strategies. In this study, we systematically evaluated the infectivity, replication kinetics, and serial passage stability of CMNV in eight cell lines of different origins, including three insect cell lines (Sf21, C6\/36, and S2) and five fish cell lines (EMRB, EPC, ZF4, TiB, and GiCB). Based on CPE observations and viral RNA replication profiles across three blind passages, cell lines exhibiting both obvious CPE and viral RNA replication in at least two passages were classified as susceptible, including EPC (P1 and P3), TiB (P1 and P3), and S2 (P1-P3). ZF4 (P1-P3) and C6\/36 (P1 and P3), despite lacking obvious CPE, supported viral RNA replication across multiple passages and were therefore tentatively designated as atypically susceptible. In contrast, Sf21 and EMRB showed no detectable viral RNA replication in any of the three passages, while GiCB supported replication only at P1; these three cell lines were accordingly classified as non-susceptible. These findings identify multiple non-shrimp cell lines that support stable CMNV replication at the RNA level and serial passage, providing useful experimental systems for studies of viral pathogenesis, host interactions, diagnostic development, and antiviral product screening.<\/jats:p>","DOI":"10.3389\/fmicb.2026.1858574","type":"journal-article","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T05:38:35Z","timestamp":1785908315000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Insect and fish cell lines support replication and serial passage of covert mortality nodavirus"],"prefix":"10.3389","volume":"17","author":[{"given":"Zhaoxi","family":"Wang","sequence":"first","affiliation":[{"name":"State Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences","place":["Qingdao, Shandong, China"]},{"name":"Laboratory for Marine Fisheries Science and Food Production Processes, Laoshan Laboratory","place":["Qingdao, Shandong, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhanqing","family":"Sun","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences","place":["Qingdao, Shandong, China"]},{"name":"Laboratory for Marine Fisheries Science and Food Production Processes, Laoshan Laboratory","place":["Qingdao, Shandong, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tingting","family":"Xu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences","place":["Qingdao, Shandong, China"]},{"name":"Laboratory for Marine Fisheries Science and Food Production Processes, Laoshan Laboratory","place":["Qingdao, Shandong, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Na","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences","place":["Qingdao, Shandong, China"]},{"name":"Laboratory for Marine Fisheries Science and Food Production Processes, Laoshan Laboratory","place":["Qingdao, Shandong, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingli","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture, Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences","place":["Qingdao, Shandong, China"]},{"name":"Laboratory for Marine Fisheries Science and Food Production Processes, Laoshan Laboratory","place":["Qingdao, Shandong, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,8,5]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"860","DOI":"10.1128\/jvi.46.3.860-870.1983","article-title":"Analysis of extracellular West Nile virus particles produced by cell cultures from genetically resistant and susceptible mice indicates enhanced amplification of defective interfering particles by resistant cultures","volume":"46","author":"Brinton","year":"1983","journal-title":"J. Virol."},{"key":"ref2","volume-title":"Book of Abstracts of the 9th Symposium on Diseases in Asian Aquaculture (DAA9)","author":"Flegel","year":"2014"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1016\/j.mib.2012.06.002","article-title":"Living with the enemy: viral persistent infections from a friendly viewpoint","volume":"15","author":"Goic","year":"2012","journal-title":"Curr. Opin. Microbiol."},{"key":"ref5","volume-title":"Preliminary Study on the Patho-Ecology of Two Emerging Diseases of Shrimp.","author":"Hao","year":"2020"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"537","DOI":"10.1084\/jem.108.4.537","article-title":"Studies on persistent infections of tissue cultures. I. General aspects of the system","volume":"108","author":"Henle","year":"1958","journal-title":"J. Exp. Med."},{"key":"ref7","doi-asserted-by":"publisher","first-page":"6302","DOI":"10.1128\/JVI.76.12.6302-631-.2002","article-title":"A single mutation in the E2 glycoprotein important for neurovirulence influences binding of sindbis virus to neuroblastoma cells","volume":"76","author":"Lee","year":"2002","journal-title":"J. Virol."},{"key":"ref8","doi-asserted-by":"publisher","first-page":"141","DOI":"10.1007\/s12250-020-00230-5","article-title":"On the calculation of TCID50 for quantitation of virus infectivity","volume":"36","author":"Lei","year":"2021","journal-title":"Virol. Sin."},{"key":"ref9","doi-asserted-by":"publisher","first-page":"65","DOI":"10.19663\/j.issn2095-9869.20180420003","article-title":"Molecular epidemiological survey of covert mortality nodavirus (CMNV) in cultured crustaceans in China in 2016~2017","volume":"40","author":"Li","year":"2019","journal-title":"Prog. Fish. Sci."},{"key":"ref10","doi-asserted-by":"publisher","first-page":"892","DOI":"10.1038\/s41564-026-02266-x","article-title":"An emerging human eye disease is associated with aquatic virus zoonotic infection","volume":"11","author":"Liu","year":"2026","journal-title":"Nat. Microbiol."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"652","DOI":"10.1099\/jgv.0.000731","article-title":"Experimental vertical transmission of covert mortality nodavirus in Exopalaemon carinicauda","volume":"98","author":"Liu","year":"2017","journal-title":"J. Gen. Virol."},{"key":"ref12","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1016\/j.jip.2018.03.011","article-title":"Vectors and reservoir hosts of covert mortality nodavirus (CMNV) in shrimp ponds","volume":"154","author":"Liu","year":"2018","journal-title":"J. Invertebr. Pathol."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"904358","DOI":"10.3389\/fmicb.2022.904358","article-title":"Investigation of pathogenic mechanism of covert mortality Nodavirus infection in Penaeus vannamei","volume":"13","author":"Liu","year":"2022","journal-title":"Front. Microbiol."},{"key":"ref15","first-page":"221","article-title":"Multiplication of the shrimp baculovirus HHNBV with primary cell cultures from lymphoid organ of Penaeus chinensis","volume":"16","author":"Miao","year":"2000","journal-title":"Chin. J. Biotechnol."},{"key":"ref16","doi-asserted-by":"publisher","first-page":"22","DOI":"10.1007\/s12560-024-09632-0","article-title":"Navigating uncertainties in RT-qPCR and infectivity assessment of norovirus","volume":"17","author":"Mirmahdi","year":"2025","journal-title":"Food Environ. Virol."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"445","DOI":"10.1016\/j.aquaculture.2016.06.044","article-title":"Development of a TaqMan real-time RT-PCR assay for detection of covert mortality nodavirus (CMNV) in penaeid shrimp","volume":"464","author":"Pooljun","year":"2016","journal-title":"Aquaculture"},{"key":"ref18","doi-asserted-by":"publisher","first-page":"112","DOI":"10.1016\/j.jviromet.2007.06.011","article-title":"In vitro replication of Macrobrachium rosenbergii nodavirus and extra small virus in C6\/36 mosquito cell line","volume":"146","author":"Sudhakaran","year":"2007","journal-title":"J. Virol. Methods"},{"key":"ref19","first-page":"13","article-title":"Patolog\u00edas del hepatop\u00e1ncreas en camarones marinos cultivados en Am\u00e9rica y su diagn\u00f3stico diferencial mediante histopatolog\u00eda [Hepatopancreas diseases in marine shrimp cultured in the Americas and its differential diagnosis by histopathology]","volume":"50","author":"Varela-Mej\u00edas","year":"2018","journal-title":"AquaTIC"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"30","DOI":"10.3390\/v18010030","article-title":"Hidden in Plain Sight: Alphavirus Persistence and its Potential for Driving Chronic Pathogenesis","volume":"18","author":"Villanueva Guzman","year":"2026","journal-title":"Viruses"},{"key":"ref21","volume-title":"Preliminary Investigation on Host Jumping of Covert Motality Nodavirus (CMNV).","author":"Wang","year":"2020"},{"key":"ref22","doi-asserted-by":"publisher","first-page":"737161","DOI":"10.1016\/j.aquaculture.2021.737161","article-title":"Histopathological study of covert mortality nodavirus infection in sea cucumber (Apostichopus japonicus)","volume":"545","author":"Wang","year":"","journal-title":"Aquaculture"},{"key":"ref23","doi-asserted-by":"publisher","first-page":"636","DOI":"10.3390\/v13040636","article-title":"First report on natural infection of Nodavirus in an Echinodermata, sea cucumber (Apostichopus japonicas)","volume":"13","author":"Wang","year":"","journal-title":"Viruses"},{"key":"ref24","author":"WOAH","year":"2023"},{"key":"ref25","doi-asserted-by":"publisher","first-page":"789128","DOI":"10.3389\/fmars.2022.789128","article-title":"Investigation on natural infection of covert mortality nodavirus in large yellow croaker, Larimichthys crocea","volume":"9","author":"Xu","year":"2022","journal-title":"Front. Mar. Sci."},{"key":"ref26","doi-asserted-by":"publisher","first-page":"670831","DOI":"10.3389\/fmars.2021.670831","article-title":"Natural infection of covert mortality nodavirus in small yellow croaker in coastal water","volume":"8","author":"Xu","year":"2021","journal-title":"Front. Mar. Sci."},{"key":"ref27","doi-asserted-by":"publisher","first-page":"101238","DOI":"10.1016\/j.aqrep.2022.101238","article-title":"Cases report of covert mortality nodavirus infection in indoor farming Penaeus vannamei","volume":"25","author":"Yao","year":"2022","journal-title":"Aquac. Rep."},{"key":"ref28","doi-asserted-by":"publisher","first-page":"2700","DOI":"10.1099\/vir.0.070078-0","article-title":"A new nodavirus is associated with covert mortality disease of shrimp","volume":"95","author":"Zhang","year":"2014","journal-title":"J. Gen. Virol."},{"key":"ref29","doi-asserted-by":"publisher","first-page":"113","DOI":"10.1016\/j.virusres.2017.03.013","article-title":"Prevalence and distribution of covert mortality nodavirus (CMNV) in cultured crustacean","volume":"233","author":"Zhang","year":"2017","journal-title":"Virus Res."},{"key":"ref30","doi-asserted-by":"publisher","first-page":"195","DOI":"10.19663\/j.issn2095-9869.20230601002","article-title":"Investigation of the prevalence of covert mortality nodavirus (CMNV) in shrimp from 2021 to 2022","volume":"45","author":"Zhao","year":"2024","journal-title":"Prog. Fish. Sci."},{"key":"ref31","doi-asserted-by":"publisher","first-page":"743834","DOI":"10.1016\/j.aquaculture.2025.743834","article-title":"Construction of a plasmid-based reverse genetics system for the rescue of shrimp infectious covert mortality nodavirus (CMNV) and use in deciphering the role of its B2 protein in regulating its cross-species infectivity","volume":"618","author":"Zhao","year":"2026","journal-title":"Aquac."}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2026.1858574\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T05:38:37Z","timestamp":1785908317000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2026.1858574\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,5]]},"references-count":29,"alternative-id":["10.3389\/fmicb.2026.1858574"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2026.1858574","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,5]]},"article-number":"1858574"}}