抗大口黑鲈弹状病毒发酵中草药的筛选及其效果评价  

SCREENING AND EVALUATION OF ANTI-MICROPTERUS SALMOIDES RHABDOVIRUS FERMENTED CHINESE HERBS MEDICINE

作　　者：曾荣博 刘爽 张玉军 李陈 余艳枝 李元平 肖运才[4] 刘学芹[1,2] ZENG Rong-Bo;LIU Shuang;ZHANG Yu-Jun;LI Chen;YU Yan-Zhi;LI Yuan-Ping;XIAO Yun-Cai;LIU Xue-Qin(National Key Laboratory of Agricultural Microbiology,College of Fisheries,Huazhong Agricultural University,Wuhan 430070,China;Hubei Engineering Technology Research Center for Aquatic Animal Diseases Control and Prevention,Wuhan 430070,China;Hubei Huada Real Science&Technology Co.,Ltd,Wuhan 430070,China;College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]华中农业大学水产学院,农业微生物资源发掘与利用全国重点实验室,武汉430070 [2]湖北省水生动物病害防控工程技术研究中心,武汉430070 [3]湖北华大瑞尔科技有限公司,武汉430070 [4]华中农业大学动物医学院,武汉430070

出　　处：《水生生物学报》2024年第3期479-487,共9页Acta Hydrobiologica Sinica

基　　金：国家重点研发计划(2022YFF1000303)资助。

摘　　要：为筛选出能够对MSRV具有抑制效果的发酵中草药,研究通过MTT法检测8种发酵中草药(L1、L2、L3、L4、L5、X1、X2和ZHS)对EPC细胞的毒性作用,结果表明L1、L2、L3、X1、X2和ZHS未对EPC细胞造成明显的损伤和毒性,L4和L5在4 mg/mL时对细胞造成损害;通过荧光定量PCR法(qRT-PCR)筛选出X1药物(主要成分为板蓝根、鱼腥草、南芪和杏仁)对MSRV有良好的抗病毒效果;在细胞水平上通过qRT-PCR法发现X1药物能够抑制MSRV的感染;空斑实验结果表明X1药物可以降低约10倍MSRV的病毒滴度。在鱼体水平上设置空白对照组、药物对照组、攻毒组、预防组和治疗组,评价X1药物对MSRV的抗病毒效果,结果表明经X1药物处理后,治疗组可以将大口黑鲈的存活率提高20%;通过qRT-PCR法检测大口黑鲈组织中病毒载量,结果表明X1药物显著抑制MSRV在肝脏、脾脏组织中的复制;通过组织病理学观察发现X1药物可以抑制MSRV引起的在肝脏、脾脏组织病变。研究为发酵中草药在水产养殖中的应用提供新的思路,为开发抗MSRV药物提供参考依据。In order to screen fermented Chinese herbs medicine that can have an inhibitory effect on MSRV.In this study,the toxic effects of eight fermented herbs(L1,L2,L3,L4,L5,X1,X2 and ZHS)on EPC cells were detected by MTT method.The results showed that L1,L2,L3,X1,X2 and ZHS did not cause significant damage and toxicity to EPC cells.However L4 and L5 exhibited cell damage at a concentration of 4 mg/mL.The results were analyzed by fluorescence quantitative PCR method(qRT-PCR)screened that X1 drug(the main components were Isatidis Radix,Isatidis Radix,Radix Astragali and Almond)had good antiviral effect on MSRV;X1 drug was found to inhibit MSRV infection by qRT-PCR at the cellular level;the results of empty spot assay showed that X1 drug could reduce the virus titer of MSRV about 10 times.To evaluate the the antiviral effects of X1 drug on MSRV at the fish level,experimental groups were established including a blank control group,drug control group,attack group,prevention group and treatment group.The results showed that the treatment group could increase the survival rate of largemouth bass by 20%after treatment with X1 drug.Furthermore,qRT-PCR analysis of tissue samples demonstrated that the X1 drug notably suppressed MSRV replication in liver and spleen.The histopathological observation showed that X1 drug could inhibit MSRV-induced lesions in liver and spleen tissues.This study provides new ideas for the application of fermented herbs in aquaculture and provides a reference basis for the development of anti-MSRV drugs.

关 键 词：大口黑鲈弹状病毒 发酵中草药 抗病毒 大口黑鲈 

分 类 号：S941.4[农业科学—水产养殖]