仔猪副伤寒活疫苗中耐热保护剂应用相关参数的研究

发布时间：2018-11-09 09:18

【摘要】：【目的】研究并确定耐热保护剂在仔猪副伤寒活疫苗中应用的各参数,为制备细菌耐热保护剂活疫苗提供参考。【方法】将耐热保护剂与制苗菌液等体积混合,分别采用3种常用冻干曲线(1、2、3)进行冻干,抽样检测确定耐热保护剂配套冻干曲线。在确定合适冻干曲线的基础上,进一步研究耐热保护剂与不同菌龄菌液(发酵培养12、15、18和21 h)、不同浓度菌液(菌液终浓度分别为3×1010、5×1010和7×1010 CFU/m L)、不同感作时间(分别作用0、24和48 h)、耐热保护剂的不同保存时间(2-8°C分别保存0、10、20、30和40 d),以及不同分装量(2、3和4 m L)等多种不同参数对疫苗质量的影响。【结果】配套冻干曲线研究表明,曲线2冻干的产品性状、活菌存活率与耐老化指标效果最好;菌龄研究表明,37°C发酵培养18 h(位于对数生长末期或稳定前期),其冻干菌存活率达78%-81%,优于其它时间;配苗试验表明,菌液适宜终浓度为3×1010-5×1010 CFU/m L;最适感作时间为2-8°C感作24 h,冻干菌存活率可达85.3%-90.5%;耐热保护剂保存期试验表明,2-8°C保存40 d与0 d(配制当日)的保护剂冻干效果基本一致;配苗分装量试验表明,7 m L西林瓶中分装3 m L或4 m L,其冻干菌存活率与2 m L基本一致,但耐老化试验中活菌存活率比2 m L略高。【结论】收获发酵培养18 h的菌液、配苗终浓度采用3×1010-5×1010 CFU/m L,使用2-8°C保存40 d内的耐热保护剂,让保护剂与制苗用菌液2-8°C感作24 h,采用3-4 m L进行定量分装,按曲线2冻干为制备仔猪副伤寒耐热保护剂活疫苗的适合参数。
[Abstract]:[objective] to study and determine the parameters of the application of heat-resistant protective agent in live vaccine of piglet paratyphoid fever, and to provide a reference for the preparation of live vaccine of bacterial heat-resistant protectant. Three kinds of freeze-drying curves (1 ~ 2 ~ 3 ~ 3) were used for freeze-drying, and the freeze-drying curves of heat-resistant protectant were determined by sampling test. On the basis of determining the suitable freeze-drying curve, the heat resistance protectant and the bacteria solution of different ages were further studied. The final concentration of the bacterial solution was 3 脳 10 10 5 脳 1010 and 7 脳 1010 CFU/m L), respectively. The final concentrations of the bacterial solution were 3 脳 10 10 5 脳 1010 and 7 脳 1010 CFU/m L), for 12 ~ 15 ~ 15 ~ 18 and 21 h), respectively. Storage time of different sensitive incubation time (0: 24 and 48 h), respectively) (2-8 掳C) for storage of 0 10, 10, 20, 30 and 40 d), respectively The effect of different dosages (2t3 and 4 m L) on vaccine quality was studied. [results] the results showed that curve 2 had the best effect on product characters, living bacteria survival rate and aging resistance index. The results of culture at 37 掳C for 18 h (located at the end of logarithmic growth or at the early stage of stabilization) showed that the survival rate of freeze-dried bacteria was 78 to 81, which was superior to that of other time, and the optimum final concentration of bacterial solution was 3 脳 10 ~ (-5) 脳 1010 CFU/m / L; The optimum incubation time was 2-8 掳C for 24 hours, the survival rate of freeze-dried bacteria could reach 85.3% and 90.5%, and the test of preservation period of heat-resistant protectant showed that the lyophilization effect of 2-8 掳C for 40 days was basically the same as that for 0 days. The results of seedling matching test showed that the survival rate of lyophilized bacteria was basically the same as that of 2 mL. But the survival rate of live bacteria in aging tolerance test was slightly higher than that in 2 mL. [conclusion] the final concentration of seedling was 3 脳 10 ~ (-5) 脳 1010 CFU/m / L and stored at 2-8 掳C for 40 days. For 24 h, the protective agent and the inoculation liquid were divided quantitatively with 3-4 mL, and freeze-dried according to curve 2 was used as the suitable parameter for the preparation of live vaccine of paratyphoid heat resistance protective agent in piglets.
【作者单位】：中国兽医药品监察所;北京中海生物科技有限公司;
【基金】：国家重点研发计划项目(No.2016YFD0500902) 北京市科委企业提升计划项目(No.Z121102002812080)~~
【分类号】：S859.797

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