抗迟钝爱德华氏菌卵黄抗体制备及其抑菌、吞噬调理活性分析
PDF下载 (7445)顾叶华,李登峰 *,周永强.抗迟钝爱德华氏菌卵黄抗体制备及其抑菌、吞噬调理活性分析[J].宁波大学学报(理工版),2018,31(5):1-7.DOI:
GU Ye-hua,LI Deng-feng *,ZHOU Yong-qiang.Preparation of IgY and its antibacterial, phagocytize opsonize effects against Edwardsiella tarda[J].Journal of Ningbo University(Natural Science & Engineering Edition),2018,31(5):1-7.DOI:
| Title: | Preparation of IgY and its antibacterial, phagocytize opsonize effects against Edwardsiella tarda |
| 作者: | 顾叶华, 李登峰 *, 周永强 |
| Author(s): | GU Ye-hua, LI Deng-feng *, ZHOU Yong-qiang |
| 关键词: | 迟钝爱德华氏菌; 卵黄抗体; 制备; 抑菌活性; 吞噬调理 |
| Keywords: | Edwardsiella tarda; IgY; preparation; bacteriostatic activity; phagocytize opsonize |
| 分类号: | S948 |
| 文献标识码: | A |
| 摘要: | 迟钝爱德华氏菌(Edwardsiella tarda)是常见人畜共患病病原. 为获得高效抗E. tarda卵黄抗体(Immunoglobulin of Yolk, IgY), 将纯培养的迟钝爱德华氏菌灭活, 免疫蛋鸡, 由蛋黄纯化IgY, 用间接ELISA法检测IgY效价. 对制备的IgY进行了抑菌活性检测和动物保护实验: 分别将抗E. tarda IgY、普通鸡蛋IgY与E. tarda (2×103 CFU·mL-1)孵育, 涂平板, 菌落计数; 分别将抗E. tarda IgY、普通鸡蛋IgY与E. tarda (2×108 CFU·mL-1)等体积混合后注射克氏原螯虾(Procambarus clarkia), 记录各组成活率. 检测抗E. tarda IgY的吞噬调理活性: 分别将抗E. tarda IgY、普通鸡蛋IgY与E. tarda (103 CFU·mL-1)及小鼠血细胞混合后, 接种平板培养、计数. 结果发现: 制备的IgY质量浓度为8.8mg?mL-1, 每个鸡蛋可以获得IgY约130mg, 效价为1:10240; 体外相对抑菌率为86.6％; 对克氏原螯虾的平均保护率为66.7％; 能促进细胞吞噬E. tarda, 相对吞噬率为79.8％. |
| Abstract: | Edwardsiella tarda is responsible for causing bacterial disease in variety of animals (reptiles, amphibians, and fishes, including catfish, turbot and eels). To prepare specific egg yolk antibody (IgY) against it, laying hens were immunized with purified and inactivated E. tarda for three times. IgY were separated and purified from the yolk of the eggs collected one month after injection by acetic acid buffer dilution, salting-out with ammonium sulfate, and dialysis with PBS. Titer of the IgY were detected by indirect enzyme-linked immunosorbent assay. The concentration and the purity of the IgY were detected by OD value and SDS-PAGE. Results show that concentration of purified IgY is 8.8 mg·mL-1, the mean production from each egg is 130 mg and the titer is 1:10240. Phagocytize opsonize effects, bacteriostasis in vivo & in vitro of the prepared IgY were measured. Rate of bacteriostasis in vitro experiment is 86.6％; The IgY can rise the phagocytize opsonize effect rate to 79.8％; IgY can protect artificially infected crayfish Procambarus clarkia and reduce mortality rate from 100％ to 66.7％. |
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| 备注/Memo: | 收稿日期: 2018－01－23. 宁波大学学报（理工版）网址: http://journallg.nbu.edu.cn/ 基金项目: 国家海洋局开放基金（HY201602）; 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）; 宁波大学王宽诚幸福基金. 第一作者: 顾叶华（1991－）, 女, 江苏泰州人, 在读硕士研究生, 主要研究方向: 水产动物病害防治. E-mail: 1040267973@qq.com *通信作者: 李登峰（1975－）, 女, 湖南双峰人, 副教授, 主要研究方向: 水产动物病害防治. E-mail: lidengfeng@nbu.edu.cn 宁波大学学报（理工版）网址：http://journallg.nbu.edu.cn/ |
