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目的初步建立嗜水气单胞菌(Aeromonas hydrophila)和温和气单胞菌(Aeromonas sobria)二联灭活疫苗的生产工艺。方法用50 L发酵罐培养嗜水气单胞菌和温和气单胞菌,确定发酵培养时间、甲醛灭活浓度及与佐剂乳化比例,对制备的疫苗进行质量检测,并分析佐剂与无佐剂二联灭活疫苗的免疫效果。结果 50 L发酵罐培养嗜水气单胞菌和温和气单胞菌菌株10～12 h时,A650值达峰值;0.3%的甲醛于37℃灭活24 h,可完全灭活菌液;以灭活菌液与佐剂1∶1.5比例混合较合适;制备的佐剂二联灭活疫苗各项质量指标均合格,免疫保护效果优于无佐剂二联灭活疫苗。结论已初步建立嗜水气单胞菌和温和气单胞菌二联灭活疫苗的生产工艺。
Objective To establish the production process of Aeromonas hydrophila and Aeromonas sobria inactivated vaccine. Methods Aeromonas hydrophila and Aeromonas sobria were cultured in a 50 L fermenter. The fermentation culture time, the formaldehyde inactivation concentration and the ratio of emulsification to the adjuvant were determined. The quality of the prepared vaccine was tested, and the adjuvant and no Adjuvant divalent inactivated vaccine immune effect. Results A650 peaked at 10-12 h after culturing Aeromonas hydrophila and S. aeruginosa strains in a 50 L fermentor. When 0.3% formalin was inactivated at 37 ° C for 24 h, the bacterial liquid was completely inactivated. Inactivated bacilli and adjuvant 1: 1.5 ratio of mixing more appropriate preparation of adjuvant duplex inactivated vaccine quality indicators are qualified, the immune protection effect is better than without adjuvant inactivated vaccine. Conclusion The production process of Aeromonas hydrophila and Aeromonas sobria inactivated vaccine has been initially established.