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目的建立鳜致病性嗜水气单胞菌灭活疫苗原液的生产工艺。方法分别采用摇瓶和5L发酵罐于28℃培养嗜水气单胞菌GYK1株,测定不同时间菌液的A650值;培养菌液添加不同浓度的甲醛液,检测不同时间的灭活效果;依次采用摇瓶、种子罐、发酵罐培养,甲醛灭活,制备灭活疫苗原液,并进行安全性和效力试验。结果摇瓶培养时,培养菌液18h进入生长稳定期,24h左右达高峰;用5L发酵罐培养时,培养菌液8h即进入生长稳定期,10h左右达高峰;0.30%的甲醛,37℃,24h可完全灭活菌液,且甲醛残留量较低;制备的灭活疫苗原液安全性良好,效力合格。结论初步建立了鳜致病性嗜水气单胞菌灭活疫苗原液的生产工艺。
Objective To establish the production process of the pathogenic Aeromonas hydrophila inactivated vaccine. Methods Aeromonas hydrophila GYK1 strain was cultured in shake flask and 5L fermentor at 28 ℃, respectively. The A650 value of bacteria liquid was determined at different time. Different concentrations of formaldehyde solution were added into culture broth to detect inactivation effect at different times. The shake flask, seed tank, fermenter culture and formaldehyde inactivation were used to prepare the inactivated vaccine stock solution, and the safety and efficacy tests were carried out. Results When cultured in shake flasks, the culture broth was stable at 18h and peaked at 24h, and reached the peak at about 10h after culturing in 5L fermenter for 8h, reached the peak at 10h, 0.30% formaldehyde, 37 ℃, The bacterial liquid could be completely inactivated in 24h and the formaldehyde residue was low. The prepared inactivated vaccine stock solution was safe and qualified. Conclusion The production process of the pathogenic Aeromonas hydrophila inactivated vaccine was initially established.