Isolation,identification and phylogenetic analysis of a ranavirus isolated from Rana grylio
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Affiliation:

1.College of Veterinary Medicine,Sichuan Agricultural University,Chengdu 611130,China;2.College of Animal Science and Technology,Sichuan Agricultural University,Chengdu 611130,China

Clc Number:

Q943

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    Abstract:

    To explore the etiology of a serious infectious disease affecting Rana grylio at a farm in Sichuan Province,pathological examinations and pathogen isolation were conducted on the diseased frogs.The isolated pathogen was identified using transmission electron microscopy,PCR detection,and phylogenetic analysis.External clinical signs included limb swelling and ulcerations on the body surface.The dead frog exhibited an enlarged liver,spleen and kidney.Histopathological observations revealed that the liver,spleen,kidney and other organs exhibited damage characterized by significant degeneration and necrosis foci.The carp epithelioma papulosum cyprini (EPC) cell line,inoculated with diseased samples at 25 ℃,demonstrated typical cytopathic effects (CPE) after four days,including rounding and shedding,with a TCID50 of 104.12/0.1 mL.Transmission electron microscopic observation showed that the virus exhibited a regular hexagonal shape with a crystalline array of its capsid,and the diagonal diameter measured approximately 165 nm.PCR by using specific primers for the major capsid protein (MCP) gene was performed on the internal tissues of infected frogs and on EPC cells with CPE,yielding positive results.Phylogenetic analysis based on MCP gene sequences indicated that the sequences of isolated virus shared over 99% identity with ranavirus and was classed as FV3-like.This study confirmed that the isolated virus,designated TSL210813,is a ranavirus,responsible for the disease observed in R. grylio.

    Fig.1 Clinical symptoms of diseased Rana grylio
    Fig.2 Histopathological lesions of diseased Rana grylio
    Fig.3 CPE produced by inoculation of EPC cells with tissue fliterate from diseased Rana grylio
    Fig.4 Electron microgragh of the ranavirus-like particles infecting EPC cells
    Fig.5 Electrophorsis of PCR product of MCP gene
    Fig.6 Phylogenetic tree of different ranavirus strains based on MCP gene sequences
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田思璐,袁羽,徐乐,欧阳萍,陈德芳,黄小丽,耿毅. Isolation,identification and phylogenetic analysis of a ranavirus isolated from Rana grylio[J]. Jorunal of Huazhong Agricultural University,2025,44(2):258-264.

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History
  • Received:February 05,2024
  • Online: April 02,2025
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