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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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摘要

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:从其野生型大肠埃希氏菌菌株和在志贺氏志贺氏菌中产生的溶原菌,自发地诱导了两种编码细胞死亡扩展毒素(Cdt)的V型编码噬菌体(Φ62和Φ125)。贮存1个月后,通过噬菌斑试验进行的传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,但是它们的持久性要比产生Cdt的大肠杆菌高得多。 Cdt噬菌体在70°C,UV辐射30分钟和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噬菌体进行了定量分析,这些结果表明Cdt噬菌体是遗传载体和环境中cdt的天然库。

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