REUSE OF PROPERLY TREATED WASTEWATER REDUCES PRESSURE ON AQUIFERS, IMPROVES SUSTAINABILITY OF CROP PRODUCTION, AND REDUCES HUNGER BY MITIGATING YIELD LOSSES FROM DROUGHT; HOWEVER, IMPROPER TREATMENT ALSO INCREASES THE RISK OF SPREADING ANTIBIOTIC RESISTANCE. TO SUSTAIN CROP PRODUCTION WITH LIMITED WATER SUPPLY, WATER SOURCES INCLUDING RECYCLED WASTEWATER, BRACKISH GROUNDWATER, AGRICULTURAL RETURN FLOW, AND PRODUCED WATER FROM INDUSTRY MUST BE CONSIDERED. THIS PROPOSAL AIMS TO INVESTIGATE THE FATE OF INTRA- AND EXTRACELLULAR ANTIBIOTIC RESISTANCE GENES (ARG) IN AGRICULTURAL REUSE WATER. OUR SHORT-TERM GOAL TO DETERMINE THE EFFECTIVENESS OF WASTEWATER TREATMENT TECHNOLOGIES IN REDUCING THE SIZE OF SURFACE WATER RESERVOIRS AND DISRUPTING THE SPREAD OF ANTIBIOTIC RESISTANCE AMONG ENVIRONMENTAL MICROBIAL COMMUNITIES WILL BE ACHIEVED THROUGH FIVE SUPPORTING OBJECTIVES:(I) ESTABLISH METHOD TO QUANTIFY ARGS IN ENVIRONMENTAL SAMPLES;(II) QUANTIFY ABUNDANCE OF ARGS IN REUSE WATER;(III) EXAMINE IMPACT OF NUTRIENTS AND SEDIMENT ON ARG PERSISTENCE IN WATER;(IV) EVALUATE REUSE WATER TREATMENT ON FATE OF ARGS;(V) DETERMINE PERSISTENCE OF ARGS IN SOIL IRRIGATED WITH REUSE WATER.WE PROPOSE A SERIES OF EXPERIMENTS TO DIFFERENTIATE INTRA- AND EXTRACELLULAR ARGS, EVALUATE NUTRIENT AND SEDIMENT DRIVERS ON PERSISTENCE OF ARGS IN WATER AND SOIL, AND ASSESS ARG FATE UNDER DIFFERENT REUSE TREATMENT TECHNOLOGIES. THE PROPOSED RESEARCH PROJECT ADDRESSES THE PROGRAM AREA PRIORITY WATER QUANTITY AND QUALITY (A1411) UNDER THE BIOENERGY, NATURAL RESOURCES, AND ENVIRONMENT PROGRAM WITH A SPECIFIC FOCUS ON ENSURING ADEQUATE NONTRADITIONAL WATER IRRIGATION PRACTICES ARE IMPLEMENTED FOR CROP AND LIVESTOCK BY EVALUATING FACTORS IMPACTING THE FATE OF BOTH INTRA- AND EXTRACELLULAR ARGS.