1.华中农业大学农业微生物资源发掘与利用全国重点实验室，武汉 430070;2.华中农业大学动物医学院，武汉 430070;3.武汉科前生物股份有限公司，武汉 430070;4.湖北洪山实验室，武汉 430070
张译升，E-mail：18753985565@163.com
陈颖钰，E-mail：chenyingyu@mail.hzau.edu.cn
S852.62
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）
- ZHANG Yisheng 1,2
- ZHANG Sen 1,2
- LIU Guoxing 1,2
- LIU Xijian 1,2
- YANG Li 3
- CHEN Jianguo 2
- GUO Aizhen 1,2,4
GUO Aizhen
National Key Laboratory of Agricultural Microbiology in Huazhong Agricultural University， Wuhan 430070， China;College of Veterinary Medicine，Huazhong Agricultural University，Wuhan 430070，China;Hubei Hongshan Laboratory， Huazhong Agricultural University，Wuhan 430070，China
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1.National Key Laboratory of Agricultural Microbiology in Huazhong Agricultural University， Wuhan 430070， China;2.College of Veterinary Medicine，Huazhong Agricultural University，Wuhan 430070，China;3.Wuhan Keqian Biology Co.，Ltd.，Wuhan 430070，China;4.Hubei Hongshan Laboratory， Huazhong Agricultural University，Wuhan 430070，China
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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.
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张译升,张森,刘国兴,刘希健,杨莉,陈建国,郭爱珍,陈颖钰.牛支原体冻干保护性物质的初步筛选[J].华中农业大学学报,2024,43(6):282-288
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- 收稿日期:2023-06-08
- 在线发布日期: 2025-01-07
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