基于响应面分析的高抗原活性鸭疫里默氏杆菌疫苗培养基的研制及其效果评价

Development and effect evaluation of Riemerella anatipestifer vaccine medium with high antigenic activity based on response surface analysis

摘要:

【背景】鸭疫里默氏杆菌广泛存在于养殖场，引起雏鸭发生传染性浆膜炎，严重危害养鸭业的发展。【目的】提高鸭疫里默氏杆菌发酵培养水平和抗原活性，为鸭疫里默氏杆菌灭活疫苗的研制提供技术指引。【方法】利用单因素及响应面的试验设计方法，针对鸭疫里默氏杆菌进行疫苗培养基的研制，并探究不同发酵时间点该菌的抗原活性，选择抗原活性最高点时制备灭活疫苗，通过动物免疫保护试验评价疫苗免疫效果。【结果】使用研制的疫苗培养基发酵培养鸭疫里默氏杆菌，其活菌数能够达到4.68×10¹⁰ CFU/mL，较市面上该菌专用的培养基提高2.29倍。该菌发酵12 h后抗原活性达到最高，在此时制备的灭活疫苗诱导小鼠产生的抗体水平显著高于商品化灭活疫苗，攻毒保护率达到了100%。【结论】本研究研制的培养基具有优异的增菌效果，可作为生产鸭疫里默氏杆菌灭活疫苗抗原的发酵培养基，疫苗生产过程中可选择菌株抗原活性达到最高时收集菌体。

关键词:鸭疫里默氏杆菌;疫苗培养基;抗原活性;响应面

Abstract:

[Background] The widespread presence of Riemerella anatipestifer in poultry farms causes contagious serositis in ducklings, which seriously endangers the development of the poultry industry.[Objective] To improve the fermentation level and antigenic activity of R. anatipestifer and provide technical guidance for the development of R. anatipestifer inactivated vaccines. [Methods] The single factor test and response surface methodology were employed to optimize the vaccine medium for R. anatipestifer. The antigenic activity of R. anatipestifer was determined at different time points of fermentation, and the inactivated vaccine was prepared at the time point with the highest antigenic activity. The immune effect of the vaccine was evaluated by animal immunization test. [Results] The viable cell count of R. anatipestifer in the developed vaccine medium reached 4.68×10¹⁰ CFU/mL, which was 2.29 times higher than that of the commercial medium for R. anatipestifer. The antigenic activity of the strain peaked after 12 h of fermentation. The inactivated vaccine prepared at this time point induced significantly higher antibody level in mice than the commercial inactivated vaccines, with 100% protection from virus attack. [Conclusion] The medium developed in this study had excellent enrichment effect and can be used for the production of antigens for R. anatipestifer inactivated vaccine. The bacteria can be collected when the antigenic activity of the selected strain reaches the highest level during the vaccine production.

Key words:Riemerella anatipestifer;vaccine medium;antigenic activity;response surface