致病性副溶血弧菌脂多糖的制备及其对牙鲆的免疫效应

利用酚水法提取的副溶血弧菌脂多糖 (lipopolysaccharide,L PS) ,经 SDS- PAGE电泳及银染色后表明这种方法提取副溶血弧菌脂多糖显示出 4 1 .3KDa,30 .7KDa,2 5 .4 KDa以及 1 1 .8KDa 4条主要脂多糖带。用此做免疫原 ,采用加强腹腔注射法免疫牙鲆 ,测定牙鲆的血清凝集抗体效价和白细胞吞噬活性。结果表明 ,免疫后的牙鲆对副溶血弧菌的血清凝集抗体效价有显著提高 ,第 3周抗体滴度达到最高值 2 5 6 ;白细胞的吞噬活性与对照组相比有明显加强 (P<0 .0 5 )。

Preparation of Vibrio parahaemolyticus Lipopolysaccharide and Immunological Analysis of the Lipopolysaccharide Against Paralichthys olivaceus

The lipopolysaccharide(LPS) was isolated by using the phenol water method from pathogen Vibrio parahaemolyticus which was isolated from sick flatfish Paralichthys olivaceus and identified according to the conventional methods. Subsequently, the component of LPS was detected with the silver stain method and Gel Pro analyzer after SDS PAGE. The results showed that the LPS contains main four bands, which are 41.3KDa, 30.7KDa, 25.4KDa and 11.8KDa,respectively. As antigens LPS is immunized to the healthy flatfish P. olivaceus by double i.p.injection. The results show that after the fish is inoculated with LPS, its agglutinating antibody titers against dead V. parahaemolyticus increases conspicuously and about 3 weeks later reaches the highest(256). The phagocytic percentage (PP) of leucocytes in the blood of the immunized increases conspicuously more than that of the control (P<0.05).