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热灭活WSSV诱导的中国明对虾(Fenneropenaeus chinensis)在不同温度下的免疫致敏反应
曹家旺1,2,3, 孔 杰1,2, 罗 坤1,2, 栾 生1,2, 曹宝祥1,2, 史晓丽1,2, 卢 霞1,2, 冯亚萍1,2,3, 王 军1,2,3, 王明珠1,2,3, 孟宪红1,2
1.农业部海洋渔业可持续发展重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071;2.青岛海洋科学与技术国家实验室 海洋渔业科学与食物产出过程功能实验室 青岛 266071;3.上海海洋大学水产与生命学院 上海 201306
摘要:
本研究拟探究经热灭活的白斑综合征病毒(White spot syndrome virus, WSSV)能否诱导中国明对虾(Fenneropenaeus chinensis)产生免疫致敏反应。将具有典型白斑综合征症状的对虾肌肉剁碎后经60℃灭活1 h,采用单尾定量口饲的方法,连续6 d投喂不同水温条件(15℃、23℃、28℃、32℃)下的实验组对虾(E15℃、E23℃、E28℃、E32℃);同时设置常温(23℃)条件下阳性对照组C23℃(投喂未经灭活的WSSV毒饵)、对照组CF23℃(只投喂商业配合饲料)。在实验第13天对存活个体进行第二次人工WSSV感染,结果显示,WSSV经60℃处理1 h可彻底失活,表现为连续投喂6 d至第二次人工感染期间无对虾死亡,而阳性对照组C23℃死亡率为100%。截止实验第19天,E15℃、E23℃、E28℃、E32℃各组存活率分别为80.41%、33.29%、8.47%、16.43%,CF23℃组的存活率为8.89%,E15℃组与其他各实验组差异极显著(P<0.01),E23℃与CF23℃、E28℃、E32℃组差异显著(P<0.05),E28℃和E32℃组差异不显著(P>0.05);对各组实验材料进行WSSV绝对荧光定量检测,结果显示,经热灭活WSSV诱导的中国明对虾二次感染WSSV后,28℃环境下病毒增殖速度最快,高温(32℃)和低温(15℃)都会不同程度抑制WSSV的增殖速度。实验表明,热灭活WSSV可以诱导中国明对虾产生免疫致敏反应,对受WSSV感染的中国明对虾具有一定的保护作用;温度与WSSV的增殖速度密切相关。
关键词:  免疫致敏  热灭活  存活率  绝对荧光定量  白斑综合征病毒
DOI:10.11758/yykxjz.20151223001
分类号:
基金项目:国家自然科学基金(31372523)、青岛海洋科学与技术国家实验室鳌山科技创新计划项目(2015ASKJ02)和泰山学者良种工程项目共同资助
Immune Priming Response Induced by Heat-Inactivated WSSV on Fenneropenaeus chinensis at Different Temperature
CAO Jiawang1,2,3, KONG Jie1,2, LUO Kun1,2, LUAN Sheng1,2, CAO Baoxiang1,2, SHI Xiaoli1,2, LU Xia1,2, FENG Yaping1,2,3, WANG Jun1,2,3, WANG Mingzhu1,2,3, MENG Xianhong1,2
1.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071;2.Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao;3.College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306
Abstract:
This study was designed to explore whether the heat-inactivated WSSV could induce immune priming response on Fenneropenaeus chinensis. During the initial 6 days, four experiment groups (E15℃, E23℃, E28℃, and E32℃) were fed with typical symptoms of WSSV prawn muscle, which was dealt with at 60℃ for 1 h using the equivalent and quantitative test method. At the same time, we set a positive control group C23℃ (fed with the unheat-inactivated WSSV muscle) and a control group CF23℃ (fed with commercial compound feed only). At the 13th day of the experiment, all shrimps were infected with WSSV again. The result showed that 60℃ could decrease the activity of WSSV absolutely, there were no dead shrimp in the initial 12 d, but the mortality of positive control group C23℃ was 100%. The survival rates of different experiment (E15℃, E23℃, E28℃, and E32℃) and control groups were 80.41%, 33.29%, 8.47% , 16.43% and 8.89%, respectively, after 19 days. The survival rate of E15℃ showed highly significant difference from other experiments (P<0.01). In addition, there was a significant difference between E23℃ experiment group and other experiment groups (P<0.05). Nevertheless, there was no significant difference between E28℃ and E32℃ groups (P>0.05). Compared to the results of absolute quantitation, it was found that the multiplication rate of WSSV was the highest under 28℃, high (32℃) and low (15℃) temperature would inhibit the multiplication of WSSV in the shrimps which accepted the subsequent rechallenge of WSSV after the heat-inactivated WSSV induced. Experimental results showed that heat-inactivated WSSV could induce immune priming response and provide some protection to the infected shrimp by WSSV. The multiplication rate of WSSV is closely related to temperature.
Key words:  Immune priming  Heat-inactivated  Survival rate  Absolute quantitation  WSSV