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作 者:阳磊[1] 汪开毓[1] 潘延乐 周燕[1] 王二龙[1] 陈德芳[1] 吉莉莉[1] 耿毅[1] 赖为民[1]
机构地区:[1]四川农业大学鱼病研究中心,四川雅安625014
出 处:《水产学报》2015年第3期429-438,共10页Journal of Fisheries of China
基 金:教育部《长江学者和创新团队发展计划》创新团队专项(IRT0848);四川省科技厅产业链(2014NZ003);四川省科技厅应用基础专项(2014JY0143)
摘 要:为优化渔用口服微球疫苗制备工艺,实验以海藻酸钠及爱德华菌灭活疫苗为原材料,采用乳化离子交联法制备爱德华菌口服微球疫苗。通过单因素实验研究海藻酸钠浓度、水油相比例、乳化剂浓度以及乳化时间对微球疫苗包封效率的影响,根据Box-Behnken中心组合实验设计,采用4因素3水平响应面法优化爱德华菌海藻酸钠微球疫苗的制备工艺。结果表明最佳制备工艺条件为海藻酸钠浓度3.5%,水油相比例2.4∶7.6,乳化剂浓度5.4%,乳化时间13 min。研究表明,在最佳工艺条件下制备的爱德华菌口服微球疫苗成球性好,粒径分布均匀,粒径(8.88±1.26)μm,跨距0.38,包封效率96.37%,具有较好抗酸性、肠溶性以及高安全性的特点。With the rapid development of aquaculture industry, bacterial diseases in fish are breaking out frequently. Antibiotics are the main methods to prevent and control the bacterial diseasesof fish,but this has been bringing some problems, such as bacterial resistance, drug residues, food safety and environmental pollution. Vaccines to control fish disease outbreaks are the most effective way. Currently, the use of natural polymer materials for carrier preparation of oral microsphere vaccine has become a hot topic. Oral microsphere vaccine can not only prevent gastrointestinal environmental damage to the vaccine, but also achieve all the growth stages of immunization of fish, and reduce the immune costs. In order to optimize the condition of preparation in microsphere vaccine. Pathogenic bacteria Edwardsiella ictaluri were killed by formalin and sodium alginate was used, and then the oral microsphere vaccine of E. ictaluri was prepared by emulsification ionic crosslinking method. Single factors including sodium alginate concentration, water-oil ratio, emulsifier concentration and emulsification time were investigated separately. On the basis of single- factor experiments, the relationships among main preparation conditions were modeled using a 4-factor,3- level Box-Behnken experimental design. The established model was analyzed by response surface methodology to obtain the optimal preparation conditions. All results showed that the optimal preparation conditions were as follows:sodium alginate concentration was 3.5 % ,water-oil ratio was 2.4: 7.6, emulsifier concentration was 5. 4% , and emulsification time was 13 min. Under these conditions, the encapsulation efficiency of microspheres was 96. 37%. It matched with the predicted encapsulation efficiency. The microspheres had good form, diameter of particles was homogeneous distribution, diameter( 8.88 ± 1.26) p,m,span 0. 38, remarkable acid resistance, enteric and high security. By response surface methodology, preparation of the oral microsphere vaccine
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