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Water is a key resource for agricultural production in the United States. Due to projected changes in water availability across the country, long-term sustainability of agricultural production may rely on finding alternatives to traditional water sources. The aim of this study was to assess farmers' opinions on the use of nontraditional water sources (e.g., agricultural runoff, treated wastewater, recycled water, produced water, untreated surface water, and brackish surface and groundwater) for agricultural activities. A survey was distributed to fanners (n = 746) in the Mid-Atlantic and Southwest regions of the United States (U.S.) about water availability and nontraditional irrigation water perceptions. Chi-square, Fisher's exact tests, f-tests, and multinomial and ordinal logistic regression analyses were conducted. Of farmers surveyed, 80% (431/543) considered the use of nontraditional water sources to be at least moderately important and 61% (444/727) would use nontraditional water if given the option. Each of the following factors individually increased the likelihood that a farmer considered nontraditional water very important for agriculture: Farmers who lived in the Southwest region compared to the Mid-Atlantic, farmers who were concerned about water availability compared with those who were not, farmers with a graduate or professional degree compared to those with less education, farmers with access to nontraditional water, and farmers with some knowledge of nontraditional water compared to those with no reported knowledge. Concern about water availability and knowledge of nontraditional water sources were significantly associated with willingness to use these water sources (p < 0.001 for both). Water quality, food safety and health risks were the main concerns regarding nontraditional water use across both regions. Willingness to use nontraditional water increased significantly if the water quality was proven to be as good or better than farmers' current water sources (63% vs. 84%; p < 0.001). Projects focused on nontraditional water use in agriculture should be regionally tailored as our data found significant differences between farmers in two distinct U.S. regions.