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Detection of Infectious Enteroviruses, Enterovirus Genomes, Somatic Coliphages, and Bacteroides fragilis Phages in Treated Wastewater

机译:在处理后的废水中检测传染性肠病毒,肠病毒基因组,体膜毛虫和脆弱拟杆菌噬菌体

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In this study, three types of treated wastewater were tested for infectious enteroviruses, the enterovirus genome, somatic coliphages, and Bacteroides fragilis phages. The aim of this work was to determine whether the presence of the two types of bacteriophages or of the enterovirus genome was a good indicator of infectious enterovirus contamination. The enterovirus genome was detected by reverse transcription-polymerase chain reaction. Infectious enteroviruses were quantified by cell culturing (BGM cells), and the bacteriophages were quantified by plaque formation on the host bacterium (Escherichia coli or B. fragilis) in agar medium. Forty-eight samples of treated wastewater were analyzed. Sixteen samples had been subjected to a secondary treatment for 8 to 12 h (A), 16 had been subjected to a secondary treatment for 30 h (B1), and 16 had been subjected to both secondary and tertiary treatments (B2). The mean concentrations of somatic coliphages were 4.9 × 104 PFU · liter?1 for treatment line A, 9.8 × 103PFU · liter?1 for B1, and 1.4 × 103 PFU · liter?1 for B2, with all the samples testing positive (100%). The mean concentrations of B. fragilis phages were 1.7 × 103 PFU · liter?1 for A (100% positive samples), 17 to 24 PFU · liter?1 for B1 (44% positive samples), and 0.8 to 13 PFU · liter?1 for B2 (6% positive samples). The mean concentrations of infectious enteroviruses were 4 most probable number of cytopathogenic units (MPNCU) · liter?1 for A (31% positive samples) and ?1 for B1 and B2 (0% positive samples). The percentages of samples testing positive for the enterovirus genome were 100% for A, 56% for B1, and 19% for B2. The percentages of samples testing positive for the enterovirus genome were significantly higher than those for infectious enteroviruses. This finding may have been due to the presence of noninfectious enteroviruses or to the presence of infectious enteroviruses that do not multiply in BGM cell cultures. However, under our experimental conditions, nondetection of the genome implies the absence of infectious viruses. There was a significant correlation between the concentration of somatic coliphages orB. fragilis phages and the presence of infectious enteroviruses or the presence of the enterovirus genome. However, the somatic coliphage concentration did not lead to fluctuations in the infectious enterovirus concentration, whereas the B. fragilis phage concentration did.
机译:在这项研究中,对三种类型的处理过的废水进行了感染性肠道病毒,肠道病毒基因组,体细胞噬菌体和脆弱拟杆菌噬菌体的测试。这项工作的目的是确定两种噬菌体或肠病毒基因组的存在是否是感染性肠病毒污染的良好指标。通过逆转录-聚合酶链反应检测肠病毒基因组。通过细胞培养(BGM细胞)定量感染性肠病毒,并通过琼脂培养基中宿主细菌(大肠杆菌或脆弱芽孢杆菌)上噬菌斑的形成来定量噬菌体。分析了四十八个处理过的废水样品。对16个样品进行了8至12小时的二次处理(A),对16个样品进行了30小时的二次处理(B1),对16个样品进行了二次和三次处理(B2)。处理线A的体细胞噬菌体平均浓度为4.9×104 PFU·升?1,对于B1为9.8×103PFU·升?1,对于B2为1.4×103 PFU·升?1,所有样品均为阳性(100 %)。脆弱类芽孢杆菌噬菌体的平均浓度对于A(100%阳性样品)为1.7×103 PFU·升?1,对于B1(44%%阳性样品)为17至24 PFU·升?1,以及0.8至13 PFU ·B2升为1(6%阳性样本)。感染性肠道病毒的平均浓度为A(31%阳性样本)的最常见细胞致病单位数(MPNCU)·升?1,而B1和B2(0%阳性样本)的?1。肠病毒基因组检测呈阳性的样品百分率:A为100%,B1为56%,B2为19%。肠病毒基因组检测呈阳性的样品百分比显着高于传染性肠病毒。该发现可能是由于存在非感染性肠病毒或存在于BGM细胞培养物中不繁殖的感染性肠病毒所致。但是,在我们的实验条件下,未检测到基因组意味着没有感染性病毒。体细胞噬菌体或B的浓度之间存在显着相关性。脆弱的噬菌体和感染性肠病毒的存在或肠病毒基因组的存在。然而,体细胞中的噬菌体浓度并没有导致传染性肠病毒浓度的波动,而脆弱的芽孢杆菌噬菌体浓度却导致了这种变化。

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