抗鳗利斯顿氏菌鸡卵黄抗体制备及其活性研究  

Preparation of IgY and its Antibacterial and Phagocytize Opsonize Effects Against Listonella anguillarum

作　　者：周永强[1] 李登峰[1] 刘联国[1] 郭安南[1] 陈炯[1] 王重彬[1] 

机构地区：[1]宁波大学应用海洋生物技术教育部重点实验室,浙江宁波315211

出　　处：《宁波大学学报（理工版）》2012年第4期1-5,共5页Journal of Ningbo University：Natural Science and Engineering Edition

基　　金：浙江省公益性技术应用研究计划项目(2011C32004);宁波市农业攻关项目(2011C10004);宁波大学人才工程项目(XR0713017);宁波市海洋蟹类产业科技创新团队项目(2011B81003);宁波大学王宽诚幸福基金

摘　　要：鳗利斯顿氏菌(Listonella anguillarum)是常见的水产动物致病菌.为获得高效抗L.anguillarum卵黄抗体(egg yolk antibodies,IgY),用纯培养的L.anguillarum制备抗原,免疫蛋鸡,联用乙酸-乙酸钠稀释、(NH4)2SO4与Na2SO4盐析、透析法由蛋黄分离纯化IgY.对制备的IgY进行了体内、外抑菌与吞噬调理活性研究:分别将抗L.anguillarum的IgY、由普通鸡蛋IgY与L.anguillarum(103CFU.mL-1)等体积混合后注射克氏原螯虾(Procambarus clarkia),计算成活率;分别将抗L.anguillarum的IgY、普通鸡蛋IgY与L.anguillarum(106CFU.mL-1)及血细胞混合后,接种平板培养、计数.结果表明,提取的IgY浓度为8.93mg.mL-1,每个鸡蛋可获得IgY约120 mg,抗体纯度较高,效价为1:20480;体外抑菌率为65%;能促进细胞吞噬L.anguillarum,相对吞噬率为75%;对克氏原螯虾的保护率为33.3%.Listonella anguillarum （Vibrio anguillarum） is responsible for causing vibriosis in numerous marine and aquatic animals. To prepare specific egg yolk antibody （IgY） against it, laying hens were immunized with purified and inactivated L. anguillarum for three times. IgY were separated and purified from the yolk of the eggs collected one month after injection by acetic acid buffer dilution method, salting-out with ammonium sulfate and sodium sulfate, and dialysis with PBS. The concentration and the purity of IgY was detected by OD value and SDS-PAGE. Titer of the IgY was detected by indirect enzyme-linked immunosorbent assay. Results show that concentration of purified IgY is 8.93 mg.mL-1, the mean production from each egg is 120 mg and the titer is 1：20480. Phagocytize opsonize effects, bacteriostasis in vivo ＆ in vitro of the prepared IgY were measured. Rate of bacteriostasis in vitro experiment is 65%; The IgY can raise the phagocytize opsonize effects rate to 75%; IgY can prolong the survival time of artificially infected crayfish Procambarus clarkia and reduce mortality rate from 100% to 66.7%. In conclusion, the prepared IgY against L. anguillarum can restrain the growth ofL. anguillarum and enhanc the phagocytosis ofL. anguillarum by blood cells.

关 键 词：鳗利斯顿氏菌 卵黄抗体 制备 抑菌活性 吞噬调理 

分 类 号：S948[农业科学—水产养殖]