基于响应面分析的高抗原活性猪水肿病大肠杆菌疫苗培养基的研制及效果评价

【背景】猪水肿病大肠杆菌引发的疾病造成了很大的危害，但现有培养基存在培养密度低的问题。【目的】研制出高抗原活性猪水肿病大肠杆菌疫苗培养基。【方法】以常用的市售猪水肿培养基为对照，通过单因素试验、爬坡试验(Plackett-Burman, PB)、响应面(Box-Behnken, BB)试验对猪水肿培养基进行响应面优化，得到猪水肿培养基最优配方。以响应面试验得到的培养基培养猪水肿病大肠杆菌，评价不同培养时间点菌株的抗原活性，制作灭活疫苗，进行动物免疫保护试验。【结果】对研制的培养基进行扩大培养验证，发现扩大培养得到的菌株活菌数可达5×10⁹ CFU/mL以上，约为对照组的2倍。制备的灭活疫苗效价可达1:140 000，并在9 h时抗原蛋白效价达到最高。【结论】本研究研制出的疫苗培养基显著提高了猪大肠杆菌菌体密度，并可提高菌体抗原活性，为猪水肿病灭活疫苗的制备提供了技术指引。

Development and effect evaluation of a vaccine culture medium with high antigenic activity for the prevention of porcine edema disease caused by Escherichia coli based on response surface methodology

[Background] Porcine edema disease caused by Escherichia coli is a major disease attacking pigs. The existing culture medium used for vaccine preparation has a low cell density.[Objective] To develop a vaccine culture medium with high antigenic activity for the prevention of porcine edema disease. [Methods] A commercially available vaccine culture medium against porcine edema disease was used as the control. The response surface methodology was employed to optimize the composition of the culture medium through single-factor experiments, Plackett-Burman (PB) design, and Box-Behnken (BB) design. The optimized culture medium was used to culture the E. coli eliciting porcine edema disease, and the antigenic activity of the culture at different time points was evaluated. In addition, the inactivated vaccine was prepared and used for animal immunization. [Results] The expanded culture of the pathogen in the developed culture medium showed that the viable cell count of the strain reached more than 5×10⁹ CFU/mL, approximately twice that of the control group. Meanwhile, the titer of the inactivated vaccine reached 1:140 000, and the antigen protein titer peaked at the time point of 9 h. [Conclusion] The developed vaccine culture medium significantly improved the cell density and antigenic activity of E. coli causing porcine edema disease, providing technical guidance for preparing inactivated vaccines against porcine edema disease.