UNS: Developing Cross-Assembly Phage As A Marker of Human Fecal Pollution in the Environment

1510925比比在《清洁水法》颁布40多年后，我国大约40%的水道仍未达到“可钓鱼和可游泳”的目标。实现这一目标的一个重大挑战是确定污染源，并具体量化人类粪便污染。最近，通过宏基因组学数据挖掘，在人类肠道中发现了一种高度丰富的噬菌体（一种细菌病毒），命名为Cross-Assembly Phage。这种噬菌体比人类肠道中所有其他已知的噬菌体加起来都要丰富，而且似乎是人类特有的。该项目将通过在阿勒格尼河的示范研究来证明交叉组装噬菌体检测的实用性，包括与其他粪便源跟踪标记物和病毒病原体的相关性；并在国际合作者的帮助下调查污水中交叉组装噬菌体的全球丰度和普遍性。此外，该项目将包括匹兹堡水微生物组项目作为教育和推广任务。在这个项目中，中学生公民科学家将取样水，包括来自匹兹堡三条河流的水，并由工程本科学生分析水样的微生物生态。这一努力将教育公民科学家和工程学本科生有关微生物学和水质的知识。卡内基科学中心已经承诺在即将到来的展览中包括一个关于这个项目的互动亭。通过开发人类特异性的、丰富的环境中人类粪便污染的病毒标记物，拟议的项目代表了污染源跟踪方面潜在的变革性的一步。除了水质领域，拟议的研究将对其他领域产生重大影响，包括食品安全和人类微生物组研究。PI完全有资格进行拟议的研究，在环境病毒学方面有丰富的经验，并且初步发表了关于交叉组装噬菌体作为污染源跟踪标记的潜力的研究。此外，该项目已承诺合作者协助完成不同的项目任务，即qPCR检测的特异性测试和国际样本收集。项目团队可以获得所有必要的资源来完成拟议的研究。拟议工作的技术和教育方面对社会都有重大的好处。污染的影响使该国的许多水道“不宜捕鱼或游泳”；在交叉组装噬菌体中开发一种改进的人类粪便污染标记将使更准确的监管和海滩关闭，以及更经济的投资来控制污染源。该项目的结果将在同行评议的文献和会议记录中以及通过外联传播。

1510925BibbyMore than four decades after the Clean Water Act, approximately 40% of our Nation's waterways still do not meet the goal of "fishable and swimmable." A significant challenge in meeting this goal is identifying sources of pollution, and specifically quantifying human fecal pollution. Recently, a highly abundant bacteriophage (a bacterial virus), named Cross-Assembly Phage, was identified in the human gut by metagenomics data mining. This bacteriophage is more abundant than all other known bacteriophages in the human gut combined, and appears to be human specific. This project will demonstrate the utility of the Cross-Assembly Phage assay through a demonstration study in the Allegheny River, including correlation with other fecal source tracking markers and viral pathogens; and investigate the global abundance and ubiquity of Cross-Assembly Phage in sewage with the assistance of international collaborators. Additionally, this project will involve the Pittsburgh Water Microbiome project as an educational and outreach task. In this project, middle-school student citizen-scientists will sample water, including water from Pittsburgh's three rivers, and the microbial ecology of the water samples will be analyzed by engineering undergraduate students. This effort will educate both citizen-scientists and undergraduate engineering students about microbiology and water quality. The Carnegie Science Center has committed to include an interactive kiosk on the project in an upcoming exhibit.The proposed project represents a potentially transformative step forward in pollution source tracking, through the development of a human-specific, abundant viral marker of human fecal pollution in the environment. In addition to the water quality field, the proposed research will have significant impact on other fields, including food safety and human microbiome research. The PI is well qualified to conduct the proposed research, with significant experience in environmental virology, as well as an initial publication investigating the potential of Cross-Assembly Phage as a pollution source tracking marker. Additionally, the project has committed collaborators to assist with distinct project tasks, namely specificity testing for the qPCR assay and international sample collection. The project team has access to all necessary resources to complete the proposed research. Both the technical and educational aspects of the proposed work have significant benefits to society. Pollution impacts have rendered many of the nation's waterways not "fishable or swimmable"; developing an improved marker of human fecal pollution in Cross-Assembly Phage will enable more accurate regulation and beach closures, as well as more economical investments to control sources of pollution. The results from this project will be disseminated both in the peer-reviewed literature and conference proceedings as well as via outreach.