| 转基因vp28蓝藻口服疫苗半数有效量测定及其对斑马鱼的安全评价 |
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| 引用本文： | 徐杨,谢京昆,李赟卉,苏永政,何培民,贾睿.转基因vp28蓝藻口服疫苗半数有效量测定及其对斑马鱼的安全评价[J].水产学报,2021,45(2):255-264. |
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| 作者姓名： | 徐杨 谢京昆 李赟卉 苏永政 何培民 贾睿 |
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| 作者单位： | 上海海洋大学,上海海洋大学,上海海洋大学,上海海洋大学,上海海洋大学,上海海洋大学 |
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| 基金项目： | 国家重点研发计划（2019YFC0312604）;上海市农委科技兴农项目（沪农科推字(2017)第1-13号）;国家高技术研究发展计划（863计划）（2014AA093506）资助;上海市大学生创新训练计划项目（S201910264057） |
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| 摘 要： | 蓝藻是对虾幼苗的天然饵料,使用转vp28基因蓝藻(Synechococcus sp. PCC 7942)直接投喂凡纳滨对虾(Litopenaeus vanname)幼苗来防治白斑综合征病毒(WSSV),可起到“药食同源”的作用。转基因蓝藻作为对抗WSSV的口服疫苗,目前面临环境释放的安全性问题。斑马鱼是最常见的模式生物之一,本研究将转vp28基因蓝藻投喂凡纳滨对虾幼苗,验证其剂量梯度抗WSSV的影响,并测定其半数有效剂量(ED50)。以该剂量为依据投喂斑马鱼后,通过测定斑马鱼体内的酶学指标、进行组织切片观察以及养殖水体中氮磷等的变化,来探究转vp28基因蓝藻对水生生物、水体环境造成的影响。研究结果表明,随着转基因蓝藻口服疫苗剂量的增加,凡纳滨对虾幼苗抗WSSV的能力也随之增强,测得ED50为0.027 g。分别使用野生型和转基因型蓝藻配合饲料投喂斑马鱼15天,投喂结束后其生长状态良好、游动正常;投喂前后身体颜色无明显差异,不同组间体长增长的差异不显著(p > 0.05)。转基因组投喂的过氧化氢酶活力稍低于空白组和野生型组,过氧化物酶活力随着投喂天数的增长而呈现降低趋势;水质监测发现总氮和总磷无较大差异,投喂蓝藻的两组氨氮均要低于空白对照组,氨氮含量的提升对斑马鱼的抗氧化能力存在一定影响;组织切片发现斑马鱼各组的肝脏、心脏组织细胞无明显差异。研究表明转vp28基因蓝藻可有效增强凡纳滨对虾抗WSSV能力,且抗病能力随着口服疫苗剂量的升高而增强;投喂斑马鱼后,斑马鱼无明显的毒理效应,且对水质影响较小;转vp28基因蓝藻作为亚单位疫苗投放对水生生物的影响较小,可为后续产业规模应用提供基础。
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| 关 键 词： | 转基因蓝藻 斑马鱼 vp28 ED50 安全性 |
| 收稿时间： | 2020/2/4 0:00:00 |
| 修稿时间： | 2020/6/19 0:00:00 |
Determination of ED50 of transgenic cyanobacteria oral vaccine and safety evaluation on Danio rerio |
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| XU Yang,XIE Jingkun,LI Yunhui,SU Yongzheng,HE Peimin,JIA Rui.Determination of ED50 of transgenic cyanobacteria oral vaccine and safety evaluation on Danio rerio[J].Journal of Fisheries of China,2021,45(2):255-264. |
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| Authors: | XU Yang XIE Jingkun LI Yunhui SU Yongzheng HE Peimin JIA Rui |
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| Institution: | Shanghai Ocean University,Shanghai Ocean University,Shanghai Ocean University,Shanghai Ocean University,Shanghai Ocean University,Shanghai Ocean University |
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| Abstract: | Cyanobacteria is a natural bait for shrimp seedlings. Directly feeding shrimp with Synechococcus-expressed vp28 to control WSSV can play a role of "medicinal and food homology". However, the problem facing the current application of transgenic cyanobacteria as a subunit vaccine is the safety of its environmental release. Danio rerio is one of the most commonly used model creatures. In this study, Synechococcus-expressed vp28 is administered to Litopenaeus vannamei to verify the effect of its dose gradient against WSSV and determine its half effective dose (ED50). In order to verify the effects of Synechococcus-expressed vp28 on aquatic organisms and water environment under ED50, this study investigated the effects of Synechococcus-expressed vp28 on aquatic organisms and the water environment investigated by measuring the enzymatic indicators in the Danio rerio, observed tissue sections and changes in nitrogen and phosphorus in the breeding water. It is found that with the increase in the dose of Synechococcus-expressed vp28 oral vaccine, the anti-WSSV efficacy of L. vannamei also increased and the measured ED50 was 0.027 g. Feeding Danio rerio by freeze-dried wild-type and transgenic cyanobacteria with the shelled-out Artemia for 15 days, the body length and color of the Danio rerio are not significantly different. The CAT in transgenic group is significantly lower than control and wild-type groups. There is no significant difference in SOD and POD between wild-type and transgenic groups. There is no significant difference in liver and heart of Danio rerio through tissue section. The ammonia nitrogen in wild-type and transgenic groups are lower than that in control, there is no significant difference between total nitrogen and total phosphorus. Studies have shown that Synechococcus-expressed vp28 can effectively enhance the resistance to WSSV of L.vannamei, and the disease resistance is enhanced with the increase of the dose subunit vaccine. The toxicological effects of Danio rerio are not obvious and had little effect on water quality after feeding by Synechococcus-expressed vp28; this subunit vaccine has less impact on aquatic organisms, which can provide a basis for subsequent industrial scale applications. |
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| Keywords: | transgenic cyanobacteria danio rerio vp28 median effective dose (ED50) safety |
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