抗嗜水气单胞菌IgY的提纯及体外抑菌效果研究

Crude extraction and purification of IgYagainst Aeromonas hydrophila and its antibacterial activity in vitro

以灭活嗜水气单胞菌(AS 1.927菌株)为抗原免疫蛋鸡,采用水稀释法粗提蛋黄液IgY和硫酸铵法纯化粗提物中IgY,研究其在上清液中的抗体提取率及其体外抑菌效果.研究表明,pH 5.0、10×水稀释法粗提后上清液中IgY抗体提取率为78.34%,33%(v/v)饱和硫酸铵法纯化后上清液中抗体提取率为64.1%,SDS-PAGE电泳显示纯化后的IgY纯度较高.pH 5.0、10×水稀释法粗提后的特异性IgY浓度达1.0 mg/mL时,嗜水气单胞菌在18 h内无生长.结果提示,用水稀释法和硫酸铵法提取蛋黄中的抗嗜水气单胞菌特异性IgY,具有简便、快捷、高效的特点,所制备的IgY活性较高,可用于嗜水气单胞菌感染引起的水生动物疾病预防和控制.

Using water-dilution to extract crude IgY from egg yolk of hens immunized with an inactivated Aeromonas hydrophila and then introducing saturated ammonium sulfate solution to purify the IgY from crude extract, this study evaluated the immunocompetence recovery and antibacterial activity of IgY in vitro. The results showed that the immunocompetence recovery were 78.34%o in the crude extraction suspension with 10×distilled water dilution at pH 5.0 and 64.1% in purification suspension with 33% （v/v） saturated ammonium sulfate solution as precipitates. The purified IgY was of high purity as shown by SDS-PAGE. The crude extraction of IgY abolished the growth of A. hydrophila at a concentration of 1.0 mg/mL within 18 hours. In conclusion, the specific IgY from immunized hens could provide a novel alternative approach to prevention and control of diseases caused by A. hydrophila infection in several aquatic species.