[Isolation and identification of channel catfish (Ictalurus punctatus) hemorrhage reovirus].
By using cell culture and virus infection methods, a new reovirus had been isolated from channel catfish (Ictalurus punctatus) suffered with severe hemorrhage and had been identified as channel catfish reovirus (CCRV) after artificial infection in fish, electron microscopy observation, physical-chemical tests, genomic SDS-PAGE analysis and sequencing. In artificial infection test, the typical symptoms of channel catfish hemorrhage as naturally occurred could be reproduced. The isolated virus could cause typical cytopathic effect in CCO and CCK cell lines. Electron microscopy observation of ultra-thin section samples of CCRV infected CCO and CCK cells revealed that the virus replicated in cytoplasm, arrayed in crystalline, and had a non-enveloped double capsid with a diameter of 60-70 nm. Frozen-thawed, 56 degrees C 1 h, chloroform and ether had no significant effects on CCRV titer, 65 degrees C 1 h could significantly inactivated the viral infectivity. The CCRV genome SDS-PAGE analysis and nuclease sensitivity test showed that the virus genome was the same as that of viruses in Aquareovirudae and consisted of 11 segments of dsRNA assigned into three classes L1, L2, L3; M1, M2, M3 and S1, S2, S3, S4, S5 with a range of size from 0.9 to 4.4 kb. The Cloning and sequencing of the CCRV S4 segment indicated the nucleic acid number of CCRV S4 was 909 bp in length, which was exactly the same as that of GCRV S4 (AF403396) and GSRV S4 (AF403407) segments. The BLAST of CCRV S4 sequence in NCBI GenBank showed that it had a 99% and 90% similarity in sequence to the GCRV S4 and GSRV S4 segments, respectively.
Zeng
Ling-Bing
LB
Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Jingzhou 434000, China. zenglingbing@gmail.com
Xu
Jin
J
Li
Yan-Qiu
YQ
Wang
Yao
Y
Xiao
Yi
Y
Fan
Yu-Ding
YD
Zhou
Yong
Y
D000818:Animals; D001483:Base Sequence; D002397:Catfishes; D002460:Cell Line; D005393:Fish Diseases / Q000821:virology*; D008969:Molecular Sequence Data; D012087:Reoviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D012088:Reoviridae Infections / Q000662:veterinary* / Q000821:virology