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目的:制备含嗜水气单胞菌被膜或全菌的PLG疫苗微粒,以草鱼为实验动物,研究疫苗微粒的免疫保护效应。方法:采用复乳挥发法制备PLG疫苗微粒,草鱼口服免疫在4周内进行3次(每次10 mg微粒),用ELISA法测定血清和肠粘液的抗体变化,保护效应用细菌攻击实验检测。结果:获得被膜和全菌的PLG疫苗微粒,蛋白含量分别为2.31%,6.12%,2种疫苗微粒均可诱导血清及肠粘液抗体应答。腹腔接种20 LD50嗜水气单胞菌后,被膜、全菌疫苗微粒组的存活率分别为46.7%和36.7%,相对存活率分别为42.9%和32.1%,接受空微粒的草鱼仅有6.7%的存活率。结论:被膜、全菌疫苗微粒对草鱼均有显著免疫保护效应,可作为预防嗜水气单胞菌感染的疫苗。
OBJECTIVE: To prepare PLG vaccine particles containing Aeromonas hydrophila membrane or whole bacteria, and study the immune protective effect of vaccine particles by using grass carp as experimental animal. Methods: The PLG vaccine particles were prepared by double emulsion volatilization. The oral immunization of grass carp was performed 3 times (10 mg particles each time) within 4 weeks. The changes of antibody in serum and intestinal mucus were detected by ELISA. The protective effect was tested by bacterial challenge. Results: The PLG vaccine particles with the membrane and whole bacteria were obtained with the protein content of 2.31% and 6.12%, respectively. Both vaccine particles induced the serum and intestinal mucus antibody responses. After intraperitoneal inoculation of 20 LD50 Aeromonas hydrophila, the survival rates of the membrane and whole-cell vaccine particle groups were 46.7% and 36.7%, respectively, and the relative survival rates were 42.9% and 32.1% respectively. Only 6.7% Survival rate. Conclusion: The capsule and whole-cell vaccine particles have significant immunoprotective effect on grass carp, which can be used as a vaccine to prevent Aeromonas hydrophila infection.