摘要
为探求牛支原体冻干菌的有效保护性物质,利用四因素四水平正交试验和单因素试验,通过检测冻干后牛支原体HB150的活菌数,筛选并评价脱脂乳、蔗糖、甘露醇、葡聚糖、海藻糖、尿素、BSA、赖氨酸等物质对牛支原体冻干后活菌率的影响,并最终确定不同保护制剂的最佳浓度及配比。结果显示,最佳的冻干保护剂配比(m/V)为:脱脂乳15%、蔗糖4.0%、甘露醇4.0%、葡聚糖3.0%、海藻糖2.0%、尿素1.5%、BSA 1.0%、赖氨酸0.3%。该配方于-40℃、12 Pa条件下冷冻干燥20 h,能够保持牛支原体的最高活菌率,达到(55.2±2.1)%。结果表明,所筛选的动干保护剂具有较好的保护作用,可以应用于牛支原体的实际冻干操作。
In order to explore the effective protective substances for freeze-drying Mycoplasma bovis(M.bovis),this study employed a four-factor,four-level orthogonal design experiment alongside a single-factor experiment.The effects of skim milk,sucrose,mannitol,glucan,trehalose,urea,BSA,lysine,and other substances on the survival rate of M.bovis were detected by measuring the number of the live M.bovis HB150 post freeze-drying and the optimal concentrations and ratios of the different protective agents were ultimately established.The ratios of the main and auxiliary components of the freeze-drying protectants were determined through orthogonal design experiments,and the final ratios of the lyophilized protectants were as follows:15%skimmed milk,4.0%sucrose,4.0%mannitol,3.0%glucan,2.0%alginate,1.5%urea,1.0%BSA,and 0.3%lysine.When freeze-drying at-40℃and 12 Pa for 20 h,this formulation maintained the highest activity rate of M.bovis,reaching(55.2±2.1)%.The results indicated that the selected dynamic dry protectants exhibited a strong protective effect and could be effectively applied in the actual freeze-drying process of M.bovis.
作者
张译升
张森
刘国兴
刘希健
杨莉
陈建国
郭爱珍
陈颖钰
ZHANG Yisheng;ZHANG Sen;LIU Guoxing;LIU Xijian;YANG Li;CHEN Jianguo;GUO Aizhen;CHEN Yingyu(National Key Laboratory of Agricultural Microbiology in Huazhong Agricultural University,Wuhan 430070,China;College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China;Wuhan Keqian Biology Co.,Ltd.,Wuhan 430070,China;Hubei Hongshan Laboratory,Huazhong Agricultural University,Wuhan 430070,China)
出处
《华中农业大学学报》
CAS
CSCD
北大核心
2024年第6期282-288,共7页
Journal of Huazhong Agricultural University
基金
宁夏回族自治区重点研发计划项目(2023BCF01038)
国家现代农业(肉牛/牦牛)产业技术体系项目(CARS-37)。
关键词
牛支原体
冷冻干燥
保护剂
正交试验
疫苗
Mycoplasma bovis
freeze drying
protective agents
orthogonal design experiments
vaccines
