鱼类迟缓爱德华氏菌肠溶口服疫苗的制备及免疫保护效应研究  

Study on the preparation and effects of the fish enteric-coated oral vaccine with Edwardsiella tarda

作　　者：魏俊利 薛淑霞 孙妍 王雪惠[3] 董学旺 孙金生[2] Wei Junli;Xue Shuxia;Sun Yan;Wang Xuehui;Dong Xuewang;Sun Jinsheng(Animal Disease Prevention and Control Center of Tianjin,Tianjin 300402,China;College of Life Science,Tianjin Normal University,Tianjin 300487,China;Tianjin Fisheries Research Institute,Tianjin 300221,China)

机构地区：[1]天津市动物疫病预防控制中心,天津300402 [2]天津师范大学生命科学学院,天津300387 [3]天津市水产研究所,天津300221

出　　处：《天津农学院学报》2024年第2期51-55,共5页Journal of Tianjin Agricultural University

基　　金：天津市科技支撑计划项目(14ZCZDNC00007);天津市农业科技成果转化项目(201502070);天津市水产研代农业技术体系创新团队项目(ITTFRS2017008)。

摘　　要：本文采用海藻酸钠作为载体,采用乳化复沉淀方法包裹迟缓爱德华氏菌(Edwardsiella tarda)全菌制备疫苗微球。制备工艺研究结果表明:3%(m/v)海藻酸钠溶液,4%(m/v)CaCl_(2)溶液,水油相比1∶2,乳化剂Span801%,搅拌速度1200 r/min为最佳制备工艺,微球包埋率达91.04%,微球平均粒径(16.51±11.87)μm。通过人工胃肠液对疫苗微球的耐受性进行分析,微球在体外人工模拟胃液和肠液中16h释放率分别为0.8%和98.1%。用表达有绿色荧光蛋白的大肠杆菌做示踪标记,口灌免疫半滑舌鳎(Cynoglossus semilaevis Gunther),疫苗能耐受胃的酸性环境影响,通过前消化道到达后肠。免疫保护效应研究表明,口腔灌注疫苗后,出现特异性和非特异性免疫应答,疫苗组在初免后第14天血清效价达到峰值1∶322;外周血白细胞吞噬率在初免后第14天达到峰值27.06%,吞噬指数在初免后第14天达到峰值1.65。初免后第28天用迟缓爱德华氏菌活菌攻毒,对照组死亡率58.82%,疫苗组死亡率13.33%,免疫保护率为77.33%。本研究优化了鱼类肠溶口服疫苗的制备工艺,疫苗免疫评价结果表明,该口服疫苗可以有效激活鱼体免疫反应,预防鱼类疾病发生。Seawater factory farming is a modern high-tech fishery with the advantage of water and energy saving.However,the diseases of the cultured fish happened frequently for the high density and intensive farming model and has caused great economic loss to the culture enterprises.Vaccination is an effective way to control the disease of cultured fish.Compared with injection and soaking,oral vaccine has the advantages of convenient inoculation,no injury to the body and good compliance of the receptor,which has become a hot spot of research.Sodium alginate is a natural macromolecule polysaccharide from the ocean.It is nontoxic,biocompatible and degradable.The degradation products can be fully absorbed by the organism.The use of sodium alginate as the carrier material to prepare the microsphere vaccine can increase the viscosity and prolong the release time of the drug.Sodium alginate is feasible to prepare the oral vaccine for fishing as a carrier for its low price,uncomplicated technology and convenient source.The study developed a new method to generate fish oral vaccine of Edwardsiella tarda by emulsfication with sodium alginate as the carrier.The results showed that the optimum preparation process of oral vaccine was 3%(m/v)sodium alginate solution,4%(m/v)calcium chloride solution,1∶2 ratios of water and oil,1%Span80 emulgator,1200 r/min stirring rate.The embedding rate was 91.04%.The mean grain size of the microballon was(16.51±11.87)μm.It is necessary to prepare the microsphere vaccine suitable for the intestinal targeted drug delivery,and the artificial simulated gastric and intestinal juice were used to analyze the tolerance of vaccine microspheres when treated by them in vitro.The results showed that the releasing rate of the microballon were 0.8%and 98.1%after dealing with simulated gastric and intestinal juice for 16 hours,respectively.Escherichia coli expressing green fluorescent protein emulsficated with sodium alginate was orally administrated to Cynoglossus semilaevis Gunther to detect the tolerance of

关 键 词：迟缓爱德华氏菌 口服疫苗 肠溶微球 免疫保护效应 

分 类 号：S942.5[农业科学—水产养殖]