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

1510925 Bibby在《清洁水法》颁布四十多年后，我国大约40%的水道仍然没有达到“可捕鱼和可游泳”的目标。“实现这一目标的一个重大挑战是确定污染源，特别是量化人类粪便污染。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），称为交叉组装噬菌体。这种噬菌体比人类肠道中所有其他已知噬菌体的总和更丰富，并且似乎是人类特异性的。该项目将通过在阿勒格尼河的示范研究来展示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;并在国际合作者的协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，该项目将涉及匹兹堡水微生物组项目作为一项教育和推广任务。在这个项目中，中学生公民科学家将对水进行采样，包括匹兹堡三条河流的水，水样的微生物生态学将由工程本科生进行分析。这一努力将教育公民科学家和本科工程专业的学生有关微生物学和水质。卡内基科学中心已承诺在即将举行的展览中包括一个互动亭项目。拟议的项目代表了一个潜在的变革性的一步，在污染源跟踪，通过开发一个人类特异性的，丰富的病毒标记的人类粪便污染的环境。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，在环境病毒学方面具有丰富的经验，以及研究交叉组装噬菌体作为污染源跟踪标记物的潜力的初步出版物。此外，该项目还委托合作者协助完成不同的项目任务，即qPCR检测试剂盒的特异性检测和国际样本采集。项目团队可以使用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利益。污染的影响已经使美国的许多水道“不适合捕鱼或游泳”;在跨大会噬菌体中开发一种改进的人类粪便污染标记物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。该项目的成果将在同行审查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.