猪丹毒疫苗效力检验攻毒冻干培养物的制备与应用研究

Preparation and Application of Erysipelothrix rhusiopathiae Challenge Culture for Potency Testing of Erysipelas Bacterins

关键词:  猪丹毒丝菌  疫苗  活菌计数  效力检验  攻毒

Key words:  E. rhusiopathiae  vaccine  plate counts  potency testing  challenge

摘要:

为提高猪丹毒疫苗效力检验结果的稳定性、一致性和工作效率。本研究冻干制备了效力检验攻毒用猪丹毒丝菌培养物，对性状、纯粹、真空度、剩余水分、活菌计数、均一性等进行检测，测定在不同温度下短期存放的活菌数，监测在低温下长期保存的活菌数和毒力，并将其应用于猪丹毒活疫苗和灭活疫苗的效力检验。结果显示，该攻毒培养物性状良好，纯粹生长，真空度合格率98.2 %，剩余水分小于3.5 %，活菌数均值为3.8×109 CFU/瓶，瓶间活菌数变异系数（C.V）为10.0 %，37℃以下温度存放2.5 ~ 3.0 h活菌数稳定，变异系数（C.V）为4.9 %，-20 ℃保存36个月活菌数为3.0×109 CFU/瓶，每只小鼠注射6 CFU活菌 5/5死亡，应用于猪丹毒灭活疫苗和活疫苗效力检验结果与常规方法一致。结果表明，该猪丹毒丝菌冻干培养物状态良好，活菌数和毒力较为稳定，能够满足猪丹毒疫苗效力检验攻毒使用，且提高了检验结果的一致性和稳定性及工作效率。

Abstract:

In order to improve stability, uniformity and efficiency of potency testing of Erysipelas Bacterins. Erysipelothrix rhusiopathiae challenge culture was prepared by freeze – drying, and its characteristics, purity, plate counts, virulence, vacuum degree, residual moisture content were determined. Its plate counts were tested when stored in different temperatures for short time，and plate counts and virulence were monitored when stored in low temperature for long time. It was applied in potency testing of Erysipelas inactivated and live vaccines. The results showed that this kind of E. rhusiopathiae challenge culture has of good characteristics and purity, its residual moisture content was under 3.5 %, the average value of plate counts was 3.8×109 CFU per bottle and the coefficient of variation of the 10 samples was 10.0 %，the plate counts did not change obviously under 37 ℃for 2.5 ~ 3.0 h，and it contained 3.0×109 CFU per bottle and changing one mice by 6 CFU could lead to 100% mortality of mouse after stored in -20℃for 36 months. It was applied in testing of Erysipelas Bacterins as challenge culture effectively, and the potency testing results were identical with the traditional culture. These data suggest that freeze – drying E. rhusiopathiae challenge culture is of good characteristics, its plate counts and virulence show a better stability, and it can be applied in potency testing of Erysipelas Bacterins and improves stability, uniformity and efficiency.