Establishment of a cell line from egg of rare minnow Gobiocypris rarus for propagation of grass carp reovirus genotype II

Affiliation
Key Lab of Fishery Drug Development, Ministry of Agriculture, Key Lab of Aquatic Animal Immune Technology, Peal River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China
Liu, Shixu;
Affiliation
Key Lab of Fishery Drug Development, Ministry of Agriculture, Key Lab of Aquatic Animal Immune Technology, Peal River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China
Wang, Yingying;
Affiliation
College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, China
Chen, Jiaming;
Affiliation
Key Lab of Fishery Drug Development, Ministry of Agriculture, Key Lab of Aquatic Animal Immune Technology, Peal River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China
Wang, Qing;
Affiliation
Key Lab of Fishery Drug Development, Ministry of Agriculture, Key Lab of Aquatic Animal Immune Technology, Peal River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China
Chang, Ouqin;
Affiliation
Key Lab of Fishery Drug Development, Ministry of Agriculture, Key Lab of Aquatic Animal Immune Technology, Peal River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China
Zeng, Weiwei;
GND
1019562625
Affiliation
Institute of Infectology, Friedrich-Loeffler-Institut (FLI), Federal Research Institute for Animal Health, Insel Riems, Greifswald, Germany
Bergmann, Sven;
Affiliation
Key Lab of Fishery Drug Development, Ministry of Agriculture, Key Lab of Aquatic Animal Immune Technology, Peal River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China
Li, Yingying;
Affiliation
Key Lab of Fishery Drug Development, Ministry of Agriculture, Key Lab of Aquatic Animal Immune Technology, Peal River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China
Yin, Jiyuan;
Affiliation
College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, China
Wen, Hong

The rare minnow, Gobiocypris rarus, is small experimental fish proven to be sensitive to Grass Carp Reovirus (GCRV) infection. In present study we established a new cell (GrE) from eggs of G. rarus. GrE cells grew well at 28 °C in M199 medium containing 10% fetal bovine serum, and has been subcultured for over 70 passages. Chromosome analysis indicated that 40% of the cells were diploid 2n = 66 while the chromosome number of the fish is 2n = 50. Viral replication in GrE cells was confirmed by transmission electron microscopy, immunofluorescence assays and virus titration experiments. GrE cells and Cyenopharyngodon idellus kidney cells were infected with two GCRV genotypes while the virus copies of GCRV II in GrE peaked at 2.25 × 10⁵ on 12th dpi. In vivo challenge experiments using GCRV I and II isolates at generations 1 and 20 indicated that GCRV II reproduce similar symptoms and histopathological changes of the disease in the rare minnow. These results indicated that GrE is permissive for GCRV genotype II propagation and can be used for pathogenesis studies and vaccine development of the predominant genotype of GCRV.

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