Home > Archive>Volume 43, Issue 6, 2024 >282-288. DOI:10.13300/j.cnki.hnlkxb.2024.06.027
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Pre-screening study on freeze-drying protectants for Mycoplasma bovis
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Affiliation:

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

Clc Number:

S852.62

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    Abstract:

    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.

    Fig.1 Number of bacteria after freeze-drying of different protectant formulations
    Fig.2 Viability of Mycoplasma bovis HB150 freeze-drying after adding mannitol （A）， sorbitol （B），and polyethylene glycol 8000 （C）
    Fig.3 Post-freeze-drying viability of Mycoplasma bovis supplemented with different levels of galactose （A），urea （B） and ascorbic acid （C）
    Fig.4 Post-freeze-drying viability of Mycoplasma bovis with the addition of BSA（A），casein （B） and NBS（C）
    Fig.5 Post-freeze-drying viability of Mycoplasma bovis with the addition of glycine （A）， lysine （B）， and arginine （C）
    Fig.6 Mycoplasma bovis HB150 strain freeze-drying material appearance
    Table 1 Freeze-drying protectant factor levels
    Table 2 Orthogonal design table for freeze-drying protectants
    Table 3 Effect of different protectant formulations on the number of viable bacteria after freeze-drying
    Table 4 Analysis of the optimal freeze-drying protectant formulation on the effect of preservation time test
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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. Pre-screening study on freeze-drying protectants for Mycoplasma bovis[J]. Jorunal of Huazhong Agricultural University,2024,43(6):282-288.

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History
  • Received:June 08,2023
  • Online: January 07,2025
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