【摘要】 为探索产气荚膜梭菌(Clostridium perfringens,Cp)的噬菌体疗法,本研究以Cp为宿主菌,在山东某地区规模化肉牛养殖场采集污水以淘选特异性噬菌体,使用透射电镜观察噬菌体形态结构,通过最佳感染复数、宿主谱、一步生长曲线、抑菌试验等对噬菌体特性进行分析。结果显示,采集的18份污水样品中成功分离到1株能够特异性识别Cp的噬菌体,噬菌斑透明无晕圈,与参试的其他细菌不发生裂解反应,最佳感染复数为0.1,在30 min进入暴发期,经测试在pH值为3～13时均保持较高的滴度,60℃处理60 min后仍能保持活性,稳定性良好。动物体内试验表明,分离获得的噬菌体能够有效抑制Cp在小鼠体内的增殖。本研究成功分离了1株噬菌体,命名为vB-CpP-Bp7,其对Cp具有良好的裂解能力、酸碱稳定性和热稳定能力,能够在体内有效降低Cp滴度,为噬菌体疗法的进一步应用提供了理论和试验依据。更多还原

【Abstract】 In order to the phage therapy for Clostridium perfringens(Cp),Cp was used as the host bacterium to screen specific phages from sewage collected from a large-scale beef cartle farm in a region of Shandong Province.Transmission electron microscopy was utilized to observe the morphological structure of the phages.Their characteristics were analyzed through the optimal multiplicity of infection(MOI),host range,one-step growth curve,and antibacterial experiments.The results showed that one phage was successfully isolated from 18 sewage samples.The plaque was transparent without a halo,exhibiting a broad host range and no lytic reaction with other tested bacteria.The optimal MOI was 0.1,and the phage entered the burst phase in 30 min.The phage maintained high titers at pH3-13 and remained active after treatment at 60℃ for 60 min,demonstrating good stability.Mice were infected with CP and treated with bacteriophage.Animal experiments showed that the isolated bacteriophage had a significant inhibitory effect on CP.The results indicated that the isolated phage,designated vB-CpP-Bp7,was a virulent phage with excellent lytic capability,acid-base stability,and thermal stability,laying a foundation for further application of this phage in phage therapy.更多还原