广西斑点叉尾鮰源嗜水气单胞菌灭活疫苗的制备

Development and Characterization of Aeromonas hydrophila Inactivated Vaccine for Ictalurus punctatus in Guangxi

本研究针对广西地区分离到的致病性嗜水气单胞菌,开发区域定制化的灭活疫苗,以实现鱼病的有效预防和控制。首先,对斑点叉尾鮰源嗜水气单胞菌在LB培养基中的生长曲线,培养基的最适pH值,以及培养基中添加蔗糖对嗜水气单胞菌生长影响进行研究;随后探索嗜水气单胞菌灭活疫苗的制备方法,并在细胞水平和斑点叉尾鮰活体水平上研究疫苗的安全性,开展疫苗对斑点叉尾鮰的免疫保护实验。结果表明嗜水气单胞菌浓度与菌液OD值间具有较好的线性关系,其回归方程为y=2E-10 x+0.1399,R 2=0.975 4。用于培养嗜水气单胞菌的LB培养基最适pH值为7.0~7.5,且在培养基中添加蔗糖能显著促进嗜水气单胞菌的生长。研究制备的嗜水气单胞菌灭活疫苗在细胞水平和鱼活体水平均安全无毒,且对斑点叉尾鮰的免疫保护率可达到75%。据此,本研究初步建立起广西斑点叉尾鮰源嗜水气单胞菌灭活疫苗的制备工艺。

In view of the pathogenic Aeromonas hydrophila isolated from Guangxi,we tried to develop regional customized inactivated vaccine to achieve effective prevention and control of fish diseases.First,the growth curve of Aeromonas hydrophila in LB medium,the optimum pH value of the medium,and the effect of adding sucrose on the growth of Aeromonas hydrophila in the medium were studied.Subsequently,the preparation method of Aeromonas hydrophila inactivated vaccine was explored,and the safety of the vaccine was studied at the cellular level and the living level of Ictalurus punctatus to carry out the immune protection experiment of the vaccine against Ictalurus nctatus.The results showed that there was a good linear relationship between the concentration of Aeromonas hydrophila and the OD value of bacterial liquid,whose regression equation was y=2E-10 x+0.1399,R 2=0.975 4.The optimum pH of LB medium for Aeromonas hydrophila was 7.0-7.5,and the addition of sucrose to the medium could significantly promote the growth of Aeromonas hydrophila.The inactivated vaccine against Aeromonas hydrophila prepared at the study was safe and non toxic at both the cellular level and the living fish level,and the immunization protection rate against Ictalurus punctatus could reach 75%.Based on this,the preparation method of inactivated vaccine against Aeromonas hydrophila from Ictalurus punctatus in Guangxi was preliminarily established.