**AWARDS ISSUED PRIOR TO JANUARY 20, 2025, WERE FUNDED UNDER PREVIOUS ADMINISTRATIONS AND MAY NOT REFLECT THE PRIORITIES AND POLICIES OF THE CURRENT ADMINISTRATION.** SURFACE WATER WORLDWIDE IS IMPACTED BY FECAL POLLUTION RESULTING FROM HUMAN AND AGRICULTURAL INPUTS THAT LEAD TO TRANSMISSION OF WATERBORNE PATHOGENS AND LOADING OF EXCESS NUTRIENTS THAT CAN CAUSE HARMFUL ALGAL BLOOMS AND DEPLETION OF DISSOLVED OXYGEN. WATERSHED MANAGERS REQUIRE ACCURATE METHODS TO IDENTIFY THE SPECIFIC SOURCES OF FECAL CONTAMINATION IN ORDER TO IMPLEMENT POLICIES AND BEST MANAGEMENT PRACTICES NECESSARY TO REMEDIATE WATER QUALITY. THE USE OF NEXT GENERATION SEQUENCING (NGS) TECHNOLOGIES HOLDS PROMISE AS AN INNOVATIVE APPROACH TO MICROBIAL SOURCE TRACKING (MST), A GROUP OF TOOLSUSED TO IDENTIFY SPECIFIC SOURCES OF FECAL CONTAMINATION. THE LONG-TERM GOAL OF THIS PROJECT IS TO DEVELOP NOVEL, HIGH-THROUGHPUT MST METHODS THAT UTILIZE NGS TO ACHIEVE INCREASED ACCURACY, RESOLUTION, SPECIFICITY, AND SENSITIVITY IN IDENTIFYING SOURCES OF FECAL CONTAMINATION TO WATERWAYS COMPARED TO EXISTING METHODS. THIS SEED PROJECT LAYS THE FRAMEWORK FOR THIS LONG-TERM GOAL AND SEEKS TO DEMONSTRATE THE FEASIBILITY OF THIS APPROACH BY (1) IDENTIFYING CANDIDATE SOURCE TRACKING TARGETS, (2) DESIGNING A CANDIDATE NGS-BASED MST METHOD OR APPROACH, AND (3) ASSESSING PERFORMANCE OF THE METHOD IN IDENTIFYING FECAL CONTAMINATION SOURCES IN MIXED LAND USE WATERSHEDS COMPARED TO TRADITIONAL MST APPROACHES. THIS WORK IS CLOSELY ALIGNED WITH THE PRIORITIES OF THE ENGINEERING FOR PRECISION CROP AND WATER MANAGEMENT PROGRAM AREA BECAUSE THIS PROJECT SEEKS TO DEVELOP AND IMPROVE ENGINEERING TECHNOLOGIES THAT CAN BE APPLIED TO PREVENT THE SPREAD OF DISEASE AND OTHER IMPACTS TO ENVIRONMENTAL WATER QUALITY FROMAGRICULTURAL SYSTEMS.