基于响应面法的高抗原活性禽多杀性巴氏杆菌疫苗培养基的研制与效果评价

Development and effect evaluation of the medium for a Pasteurella multocida vaccine with high antigenic activity based on response surface methodology

摘要:

【背景】禽多杀性巴氏杆菌(Pasteurella multocida)引发的禽霍乱疫情造成了巨大的危害，而现有培养基存在培养菌密度较低的问题。【目的】研制高抗原活性的禽多杀性巴氏杆菌疫苗培养基。【方法】通过单因素试验、Plackett-Burman试验和响应面分析方法对禽多杀性巴氏杆菌培养基的成分进行调整，并对不同发酵阶段的菌体进行免疫原性测定。最后使用该培养基培养细菌后制备疫苗并通过动物攻毒试验评价其保护效果。【结果】使用研制的培养基培养禽多杀性巴氏杆菌，活菌密度能够在6 h达到约1.84×10¹⁰ CFU/mL，增菌效果是对照培养基的2.6倍；免疫原性测定结果显示在生长平台期菌体的抗原活性最高；攻毒试验表明制备的疫苗能够很好地抵抗禽多杀性巴氏杆菌的侵袭。【结论】研制出了高抗原活性的禽多杀性巴氏杆菌疫苗培养基，为疫苗的生产奠定了基础。

关键词:禽多杀性巴氏杆菌;高抗原活性;疫苗培养基;响应面法

Abstract:

[Background] The fowl cholera caused by Pasteurella multocida causes great harm to the production, while the existing culture medium has the problem of low bacterial density. [Objective] To develop the medium for the preparation of a P. multocida vaccine with high antigenic activity. [Methods] Single factor experiment, Plackett-Burman design, and response surface methodology were employed to optimize the medium composition of P. multocida. Next, the immunogenicity of the bacteria in different fermentation phases was determined. Finally, the bacteria cultured in this medium were used to prepare the vaccine, the protective effect of which was evaluated by an animal challenge test. [Results] When the developed medium was used for the culture of P. multocida, the maximum viable count reached 1.84×10¹⁰ CFU/mL in 6 h, which was 2.6 times that of the control medium. The antigenic activity of the fermented product was the highest in the stationary phase. Challenge test showed that the vaccine prepared with this culture medium well resisted the infection of P. multocida. [Conclusion] We developed the medium for preparing a P. multocida vaccine with high antigenic activity, laying a foundation for vaccine production.

Key words:Pasteurella multocida;high antigenic activity;vaccine medium;response surface methodology