宿主菌富集空肠弯曲菌噬菌体培养条件的优化

Optimization of culture conditions for enrichment of Campylobacter jejuni phages by host bacteria

摘要:

[背景] 开发噬菌体产品是一种防控空肠弯曲菌有潜力的策略，但是面临噬菌体分离的挑战。[目的] 运用响应面法对宿主菌富集空肠弯曲菌噬菌体的培养条件进行优化。[方法] 通过单因素试验分析培养基、培养温度、培养转速、离子添加剂对噬菌体富集效果的影响，以噬菌体回收率为评价指标，采用响应面法优化了空肠弯曲菌噬菌体的富集培养条件。[结果] 在37℃条件下进行静置培养时，噬菌体富集培养效果最佳，回收率为354.12%。分离噬菌体的过程包括采样并制备滤液、宿主菌与样品滤液共培养及噬菌体分离与鉴定等环节。应用此方法从鸡粪便中分离空肠弯曲菌噬菌体，与传统的单斑法相比，噬菌体分离率提高了269.23%。[结论] 研究优化的宿主菌富集噬菌体培养方法可提高空肠弯曲菌噬菌体的分离效率，为噬菌体的研究提供思路。

关键词:噬菌体;空肠弯曲菌;分离;富集

Abstract:

[Background] The development of a bacteriophage-based product for the control of Campylobacter jejuni is a promising biocontrol strategy. However, bacteriophage isolation is a complicated challenge. [Objective] Response surface methodology was used to optimize the enrichment conditions of Campylobacter jejuni phages. [Methods] The effects of culture medium, culture temperature, shaking and ion additive on phage enrichment were analyzed through the single factor test. Based on response surface methodology (RSM), the recovery efficiency of phage was selected to evaluate enrichment culture conditions. [Results] The optimal culture conditions for enrichment were 37℃ and 0 r/min, recovery efficiency was 354.12%. The main procedures of bacteriophage isolation include sampling and sample preparation, incubation of sample filtrated with indicator strains, bacteriophage isolation and identification. Compared to the single spot method without enrichment, this enrichment method could increase the phage isolation rate by 269.23% from chicken fecal samples. [Conclusion] The optimized enrichment conditions of Campylobacter jejuni phages could improve the isolation efficiency of phages. It provides a new idea for phage isolation.

Key words:phage;Campylobacter jejuni;isolation;enrichment