새로운 betanodavirus 재편성체(reassortant)의어류 치어에 대한 병원성 분석
Pathogenicity of new reassortant betanodaviruses to various juvenile fishes
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With the recent isolation of a new betanodavirus in shellfish, Korean Shellfish Nervous Necrosis Virus (KSNNV), it has also been identified the reassortant KSNNV of two RNA segments, in which one segment is KSNNV genotype but the other one is known genotype. In this study, we confirmed that the ressortant KSNNVs obtained in previous screening study of our laboratory for betanodaviruses in shellfish were KS/RGNNV and RG/KSNNV type by performing two consecutive multiplex RT-PCR on each RNA1 and RNA2 segment (R1- and R2-discriminative multiplex two-step RT-PCR, respectively) to determine the genotype of each segment based on the size of amplicon. In the pathogenicity analysis, none of the reassortants induced specific external symptoms or mortality of VNN, but viruses of 2×104~105 copies/mg or more were detected at 14 days after injection (107 copies/fish) in brain tissues of 4 species except for crucian carp and common carp among the 6 species of juvenile fish used. In addition, the histopathological features of weak but distinct vacuole formation were also found in the brain of these infected fish, but no difference was found between the two reassortants KS/RGNNV-KG and RG/KSNNV-CM.
Abstract
With the recent isolation of a new betanodavirus in shellfish, Korean Shellfish Nervous Necrosis Virus (KSNNV), it has also been identified the reassortant KSNNV of two RNA segments, in which one segment is KSNNV genotype but the other one is known genotype. In this study, we confirmed that the ressortant KSNNVs obtained in previous screening study of our laboratory for betanodaviruses in shellfish were KS/RGNNV and RG/KSNNV type by performing two consecutive multiplex RT-PCR on each RNA1 and RNA2 segment (R1- and R2-discriminative multiplex two-step RT-PCR, respectively) to determine the genotype of each segment based on the size of amplicon. In the pathogenicity analysis, none of the reassortants induced specific external symptoms or mortality of VNN, but viruses of 2×104~105 copies/mg or more were detected at 14 days after injection (107 copies/fish) in brain tissues of 4 species except for crucian carp and common carp among the 6 species of juvenile fish used. In addition, the histopathological features of weak but distinct vacuole formation were also found in the brain of these infected fish, but no difference was found between the two reassortants KS/RGNNV-KG and RG/KSNNV-CM.
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