Isolation and identification of a new strain of hirame rhabdovirus (HIRRV) from Japanese flounder Paralichthys olivaceus
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RESEARCH
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Isolation and identification of a new strain of hirame rhabdovirus (HIRRV) from Japanese flounder Paralichthys olivaceus in China Jialin Zhang1, Xiaoqian Tang1*, Xiuzhen Sheng1, Jing Xing1 and Wenbin Zhan1,2
Abstract Background: Hirame rhabdovirus virus (HIRRV) is a rhabdovirus that causes acute hemorrhage disease in fish culture, resulting in a great economic loss in parts of Asia and Europe. Methods: In this study, we isolated a virus strain named as CNPo2015 from cultured Japanese flounder in Shandong province, China. Cell isolation, electron microscopic observation, RT-PCR detection and phylogenetic analysis were used for virus identification. Further, artificial infection experiment was conducted for virulence testing. Results: The gross signs included abdominal distension, fin reddening and yellow ascitic fluid in the abdominal cavity. Histopathological examination revealed marked cell degeneration and necrosis in the kidney. The tissue homogenates induced obvious cytopathic effects in EPC, FHM and FG cell lines. Electron microscopic observation showed the virus had a bullet-like shape with a capsule membrane. RT-PCR and sequencing analysis revealed that CNPo2015 belonged to the HIRRV with high sequence identity to HIRRV isolates. Infection experiment confirmed that the HIRRV CNPo2015 strain was virulent to flounder juveniles with a LD50 value of 1.0 × 105.9 TCID50/fish. Conclusion: In conclusion, we described the first isolation and characterization of a HIRRV from Japanese flounder in China. This will provide a candidate material for further research on the infection mechanism and preventive strategies of HIRRV. Keywords: Hirame rhabdovirus, Paralichthys olivaceus, Identification, Pathogenicity
Background Hirame rhabdovirus (HIRRV) is a single-stranded RNA virus, which belongs to the genus Novirhabdovirus within the family Rhabdoviridae. The virus was first described in cultured Japanese flounder in Japan in the early 1980s [1], from where it gradually spread to South Korea and China [2, 3]. However, due to increasing travels and rapid globalization, the outbreak of HIRRV has also been reported in part of Europe [4]. HIRRV can affect a wide range of marine fishes including Japanese * Correspondence: [email protected] 1 Laboratory of Pathology and Immunology of Aquatic Animals, Ocean University of China, No.5 Yushan Road, Qingdao 266003, China Full list of author information is available at the end of the article
flounder, stone flounder (Kareius bicoloratus), black seabream (Acanthopagrus schlegeli) and sea bass (Lateolabrax maculatus) [5]. The major clinical signs of HIRRV infection are congestion of the gonads, focal haemorrhage of the skeletal muscle and fins and accumulation of ascitic fluid [1]. Nowadays, the lack of vaccines and drugs against HIRRV highlights the urgency and significance of investigating infection mechanism and preventive strategies against HIRRV [6]. As with all novirhabdoviruses, the HIRRV genome encodes six viral proteins including nucleoprotein (N), phosp
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