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首页> 外文期刊>International Journal of Environmental Research and Public Health >Detection of Emerging and Re-Emerging Pathogens in Surface Waters Close to an Urban Area
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Detection of Emerging and Re-Emerging Pathogens in Surface Waters Close to an Urban Area

机译：市区附近地表水中新兴病原体的检测

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Current knowledge about the spread of pathogens in aquatic environments is scarce probably because bacteria, viruses, algae and their toxins tend to occur at low concentrations in water, making them very difficult to measure directly. The purpose of this study was the development and validation of tools to detect pathogens in freshwater systems close to an urban area. In order to evaluate anthropogenic impacts on water microbiological quality, a phylogenetic microarray was developed in the context of the EU project μAQUA to detect simultaneously numerous pathogens and applied to samples from two different locations close to an urban area located upstream and downstream of Rome in the Tiber River. Furthermore, human enteric viruses were also detected. Fifty liters of water were collected and concentrated using a hollow-fiber ultrafiltration approach. The resultant concentrate was further size-fractionated through a series of decreasing pore size filters. RNA was extracted from pooled filters and hybridized to the newly designed microarray to detect pathogenic bacteria, protozoa and toxic cyanobacteria. Diatoms as indicators of the water quality status, were also included in the microarray to evaluate water quality. The microarray results gave positive signals for bacteria, diatoms, cyanobacteria and protozoa. Cross validation of the microarray was performed using standard microbiological methods for the bacteria. The presence of oral-fecal transmitted human enteric-viruses were detected using q-PCR. Significant concentrations of Salmonella, Clostridium, Campylobacter and Staphylococcus as well as Hepatitis E Virus (HEV), noroviruses GI (NoGGI) and GII (NoGII) and human adenovirus 41 (ADV 41) were found in the Mezzocammino site, whereas lower concentrations of other bacteria and only the ADV41 virus was recovered at the Castel Giubileo site. This study revealed that the pollution level in the Tiber River was considerably higher downstream rather than upstream of Rome and the downstream location was contaminated by emerging and re-emerging pathogens.
机译：关于病原体在水生环境中传播的最新知识很稀少，这可能是因为细菌，病毒，藻类及其毒素往往以低浓度存在于水中，从而很难直接测量。这项研究的目的是开发和验证用于检测靠近市区的淡水系统中病原体的工具。为了评估人为因素对水微生物质量的影响，在欧盟项目μAQUA的背景下开发了系统发育微阵列，以同时检测多种病原体，并应用于来自罗马上游和下游市区附近两个不同地点的样品台伯河。此外，还检测到人肠病毒。使用中空纤维超滤方法收集并浓缩50升水。将所得浓缩物通过一系列递减的孔径过滤器进一步分级分离。从合并的滤膜中提取RNA，然后与新设计的微阵列杂交，以检测病原细菌，原生动物和有毒的蓝细菌。硅藻作为水质状况的指标，也包括在微阵列中以评估水质。芯片结果为细菌，硅藻，蓝细菌和原生动物提供了阳性信号。使用用于细菌的标准微生物学方法对微阵列进行交叉验证。使用q-PCR检测了粪便传播的人肠道病毒的存在。在Mezzocammino部位发现了高浓度的沙门氏菌，梭菌，弯曲杆菌和葡萄球菌以及戊型肝炎病毒（HEV），诺如病毒GI（NoGGI）和GII（NoGII）和人腺病毒41（ADV 41）。细菌，只有ADV41病毒在Castel Giubileo现场被回收。这项研究表明，台伯河下游的污染水平比罗马上游要高得多，而下游位置受到了新出现和重新出现的病原体的污染。

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