温度对锦鲤疱疹病毒体外培养和致病性的影响

    Effects of temperature on culture in vitro and pathogenicity of Cyprinid herpesvirus 3

    • 摘要:
      目的  证实温度对锦鲤疱疹病毒CyHV-3体外增殖和致病性的影响。
      方法  选用CyHV-3-EGFP病毒株为研究对象,采用间接免疫荧光染色法测定病毒滴度,使用荧光显微镜观察病毒体外增殖,研究病毒的热稳定性、以及温度对病毒的体外培养和体内致病性的影响。
      结果  CyHV-3在4~36 ℃条件下具有很好的稳定性,温度超过36 ℃后,病毒活力随着温度升高而降低,50 ℃时病毒活力完全丧失。感染温度对CyHV-3病毒活力影响不大,高温(37 ℃)和低温(4 ℃)时病毒都具有感染能力。但是,孵育温度明显影响病毒活力,温度高于30 ℃时,CyHV-3的增殖能力减退甚至消失。鲤鱼感染CyHV-3病毒后,低温组(15 ℃)的死亡率为26.67%,常温组(25 ℃)的死亡率为73.33%,高温组(30 ℃)和对照组的死亡率均为0。
      结论  证实了CyHV-3病毒在低温下能够增殖并且具有致病性,解释了近年来锦鲤疱疹病毒(KHVD)的暴发及流行不再受限于春秋季节的现象。

       

      Abstract:
      Objective  To confirm the effect of temperature on proliferation in vitro and pathogenicity of Cyprinid herpesvirus 3 (CyHV-3).
      Method  CyHV-3-EGFP strain was used in this study. Viral titer was measured by the indirect immunofluorescence staining assay. The proliferation of virus in vitro was observed using a fluorescence microscope. The thermal stability, and the effect of temperature on proliferation in vitro and pathogenicity in vivo of virus were studied.
      Result  CyHV-3 had good thermal stability at 4-36 ℃. The viability of CyHV-3 decreased with the increase of temperature when the temperature exceeded 36 ℃. The virus viability was completely lost at 50 ℃. The infection temperature had little effect on the viability of CyHV-3, and the virus showed infectivity at both high (37 ℃) and low (4 ℃) temperatures. However, incubation temperature significantly affected the virus viability. CyHV-3 proliferation capacity decreased or even disappeared when the temperature exceeded 30 ℃. After CyHV-3 infection, the mortality rates for the carps at low temperature (15 ℃) and room temperature (25 ℃) were 26.67% and 73.33% respectively, and the mortality rates for the carps at high temperature (30 ℃) and in the control group with no virus infection were 0.
      Conclusion  The results confirm that CyHV-3 can proliferate and has pathogenicity at low temperature, which explain the phenomenon that the outbreak and rapid spread of KHVD in recent years are no longer restricted in spring and autumn seasons.

       

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