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

<p>More 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.</p>

<p>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.</p>