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抗大口黑鲈(Micropterus salmoides)蛙虹彩病毒药效模型的构建及其抗病毒中药筛选
李秋语1,2, 黄小红2, 郝贵杰2, 胡大雁3, 张成赛2, 牛晨2, 袁雪梅2, 黄雷2, 于喆2, 姚嘉赟1,2, 杨桂连1, 姜建湖2
1.吉林农业大学动物科学与技术学院 吉林省动物微生态制剂工程研究中心 吉林长春 130118;2.浙江省淡水水产研究所 农业农村部淡水渔业健康养殖重点实验室 浙江省鱼类健康与营养重点实验室 浙江湖州 313001;3.湖州市农业科技发展中心 浙江湖州 313001
摘要:
大口黑鲈蛙虹彩病毒(LMBV)是一种严重危害大口黑鲈养殖的病害,但目前缺乏有效的防控手段。通过研究水温、鱼体大小以及攻毒剂量等条件对大口黑鲈感染蛙虹彩病毒的影响,构建体内抗病毒药效筛选模型,同时利用MTT法构建体外药物筛选模型,进而通过体外和体内药效模型,对29种中草药进行抗病毒药效评价,对筛选出药效最佳的2种中药进行体外和体内抗病毒药效评价。结果表明:水温和鱼体大小是影响体内药效模型的2个关键因子,最佳体内药效模型条件为:水温为25℃,鱼体大小20g,攻毒剂量为0.1mL 109.33 TCID50/mL。结合体内和体外药效模型结果,筛选出紫花地丁和黄连这2种中药具有较好的抗病毒效果的中草药。体内药效结果表明:黄连和地丁的添加量为0.6g/kg和1.2g/kg时,其对大口黑鲈的保护率最高均达到40%,而两者用药后病毒在肝组织的抑制率分别可达71.5%和67.0%。组织病理学结果表明,当药物使用浓度为0.6g/kg和1.2g/kg时,可有效降低因大口黑鲈蛙虹彩病毒感染所导致的肝和肾等组织的病理学损伤。上述结果表明,黄连和地丁均具有较好的抗蛙虹彩病毒的作用。
关键词:  大口黑鲈(Micropterus salmoides)  蛙虹彩病毒  抗病毒模型  紫花地丁  黄连
DOI:10.11693/hyhz20220300071
分类号:S942;Q955;S965
基金项目:湖州市公益性应用研究项目,2021GZ28号,2021GZ24号,2021GZ31号,201105007-4号;国家重点研发蓝色粮仓项目,2019YFD0900104号;湖州市乡村振兴专项,2018ZD2025号。
CONSTRUCTION OF EFFICACY MODEL AGAINST LARGEMOUTH BASS VIRUS AND SCREENING OF ANTIVIRAL CHINESE MEDICINE
LI Qiu-Yu1,2, HUANG Xiao-Hong2, HAO Gui-Jie2, HU Da-Yan3, ZHANG Cheng-Sai2, NIU Chen2, YUAN Xue-Mei2, HUANG Lei2, YU Zhe2, YAO Jia-Yun1,2, YANG Gui-Lian1, JIANG Jian-Hu2
1.Jilin Engineering Research Center of Animal Probiotics, College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China;2.Key Laboratory of Healthy Freshwater Fisheries Aquaculture, Ministry of Agriculture and Rural Affairs, Zhejiang Key Laboratory of Fish Health and Nutrition, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China;3.Huzhou Agricultural Science and Technology Development Centre, Huzhou 313001, China
Abstract:
Largemouth bass virus (LMBV) is a type of virus in Genus Ranavirus Family Iridoviridae; it often causes lethal and epidemic diseases in Micropterus salmoides in cultivation. However, at percent, there is no effective cure to LMBV infection. In this study, MTT method was used to establish an in vitro drug screening model. In addition, an in vivo pharmacodynamic model was constructed using three variables:water temperature, fish body size, and the dose of virus. Twenty-nine Chinese herbs were selected to evaluate their antiviral efficacy against the virus in the in vitro and in vivo pharmacodynamic models. Results show that water temperature and fish body size were two key factors affecting the in vivo model. The optimal pharmacodynamic model conditions were:water temperature 25℃, fish body size 20 g, and the dose of LMBV 0.1 mL 109.33 TCID50/mL. Considering the in vivo and in vitro efficacy, extracts of two Chinese herbs, i.e., Viola philippica and Coptis chinensis Franch showed good antiviral effects among 29 Chinese herbs tested. Under the concentration of V. philippica ≥ 0.6 g/kg and C. chinensis Franch ≥ 1.2 g/kg, the protection rates against LMBV reached 40%. Compared with non-treatment fish, the viral load of LMBV in V. philippica and C. chinensis Franch treatment group was reduced by 71.5% and 67.0%, respectively. Moreover, V. philippica and C. chinensis Franch could significantly relieve pathological damage in liver and kidney. Therefore, V. philippica and C. chinensis Franch can perform well against LMBV.
Key words:  Micropterus salmoides  largemouth bass virus  antiviral model  Viola philippica  Coptis chinensis Franch
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