Isolation, identification and phylogenetic analysis of a ranavirus isolated from <i>Rana nigromaculata</i>

MOU Weihao; GENG Yi; OUYANG Ping; CHEN Defang; HUANG Xiaoli; WANG Kaiyu; ZHAO Ruoxuan; BAI Minghuan

doi:10.7671/j.issn.1001-411X.201805012

Journal of South China Agricultural University > 2019 > 40(2): 40-46. > DOI: 10.7671/j.issn.1001-411X.201805012

MOU Weihao, GENG Yi, OUYANG Ping, et al. Isolation, identification and phylogenetic analysis of a ranavirus isolated from Rana nigromaculata[J]. Journal of South China Agricultural University, 2019, 40(2): 40-46. DOI: 10.7671/j.issn.1001-411X.201805012

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Isolation, identification and phylogenetic analysis of a ranavirus isolated from Rana nigromaculata

College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China

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Received Date: May 20, 2018
Available Online: May 17, 2023

Graphical Abstract

Abstract

Abstract

Objective
To explore the etiology of a serious infectious disease occured in Rana nigromaculata tadpoles in a farm of Sichuan Province.

Method
Pathological examination and virus isolation were carried out for the diseased tadpoles of R. nigromaculata. The isolated pathogen was identified by artificial infection test, transmission electron microscopy, PCR detection and phylogenetic analysis.

Result
External clinical signs included hemorrhage on body surface and swollen abdomen with yellow ascites. Based on histopathological observation, we found that liver, spleen, kidney, pancreas and other organs had damages with obvious degeneration and necrosis focus, and basophilic inclusions appeared in the cytoplasm of some pathological cells. The tissue homogenates of diseased tadpoles were inoculated into epithelioma papulosum cyprini (EPC) cells under 25 ℃ and caused typical cytopathic effect (CPE) after four days with the TCID₅₀ of 10⁸ mL^–1. In the artificial infection test of virus fluid, the tadpoles showed the symptoms similar to those observed in naturally infective tadpoles and the mortality reached 80%, which suggested that the isolated virus was the determined pathogen of this disease. Transmission electron microscopic observation showed that the virus was regular hexagon with capsule, the diagonal diameter was (135±8) nm, and virus particles arrayed in crystalline or freely in the cytoplasm. PCR examination of the MCP gene showed positive results for samples of diseased tadpoles, aquaculture water source and isolated virus. Phylogenetic analysis based on MCP gene sequences indicated that the sequence of isolated virus had above 99% similarity to ranavirus, and the isolated virus belonged to the FV3-like virus group.

Conclusion
This study confirmed that ranavirus was the causative agent of this outbreak, and the virus is named as Rana nigromaculata ranavirus (RNRV).
- ranavirus,
- isolation and identification,
- phylogenetic analysis,
- pathological lesion,
- Rana nigromaculata

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References (24)

References

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Isolation, identification and phylogenetic analysis of a ranavirus isolated from Rana nigromaculata

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