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Stability and Infectivity of Cytolethal Distending Toxin Type V Gene-Carrying Bacteriophages in a Water Mesocosm and under Different Inactivation Conditions

机译:水灭活条件下不同灭活条件下携带V型致死细胞毒素的噬菌体的稳定性和感染性

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Two cytolethal distending toxin (Cdt) type V-encoding bacteriophages (Φ62 and Φ125) were induced spontaneously from their wild-type Escherichia coli strains and from the lysogens generated in Shigella sonnei . The stability of Cdt phages was determined at various temperatures and pH values after 1 month of storage by means of infectivity tests using a plaque blot assay and analysis of phage genomes using real-time quantitative PCR (qPCR): both were highly stable. We assessed the inactivation of Cdt phages by thermal treatment, chlorination, UV radiation, and in a mesocosm in both summer and winter. The results for the two Cdt phages showed similar trends and were also similar to the phage SOM23 used for reference, but they showed a much higher persistence than Cdt-producing E. coli . Cdt phages showed maximal inactivation after 1 h at 70°C, 30 min of UV radiation, and 30 min of contact with a 10-ppm chlorine treatment. Inactivation in a mesocosm was higher in summer than in winter, probably because of solar radiation. The treatments reduced the number of infectious phages but did not have a significant effect on the Cdt phage particles detected by qPCR. Cdt phages were quantified by qPCR in 73% of river samples, and these results suggest that Cdt phages are a genetic vehicle and the natural reservoir for cdt in the environment.
机译:从它们的野生型大肠杆菌菌株和在志贺氏志贺氏菌中产生的溶原菌自发地诱导了两种细胞致死性扩张毒素(Cdt)V型编码噬菌体(Φ62和Φ125)。贮存1个月后,通过噬菌斑印迹法进行的感染性测试和实时定量PCR(qPCR)对噬菌体基因组的分析,确定了Cdt噬菌体在各种温度和pH值下的稳定性。我们通过热处理,氯化,紫外线辐射以及夏季和冬季的中膜评估了Cdt噬菌体的灭活。两种Cdt噬菌体的结果显示出相似的趋势,也与用于参考的噬菌体SOM23相似,但是它们的持久性远高于产生Cdt的大肠杆菌。 Cdt噬菌体在70°C,UV辐射30分钟和与10 ppm氯处理接触30分钟后显示最大灭活。夏季,中层的失活高于冬季,这可能是由于太阳辐射造成的。这些处理减少了感染性噬菌体的数量,但是对通过qPCR检测到的Cdt噬菌体颗粒没有显着影响。通过qPCR对73%的河流样本中的Cdt噬菌体进行了定量,这些结果表明,Cdt噬菌体是环境中cdt的遗传载体和天然库。

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