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Climate change impacts all water sources, including high quality groundwater that supplies agricultural irrigation in many regions of the United States. This study assessed groundwater level changes in the U.S. Mid-Atlantic region with a focus on cultivated areas. Trends of groundwater level were estimated using linear regression, and examined for shallow, medium, and deep depths across physiographic regions of Mid-Atlantic. A hotspot analysis was conducted to identify spatial clusters of wells with rising or declining groundwater levels. In addition, differences in the percentage of cultivated area with declining groundwater between cultivated land categories was examined at the county level. From 2002-2016, the Mid-Atlantic region had an overall decline in groundwater level (0.06 m/yr, 95% CI: 0.03, 0.09) although groundwater changes varied by physiographic regions. The Coastal Plain physiographic region was dominated by declining groundwater wells (48%) and had the most significant groundwater level declines (0.23 m/yr, 95% CI: 0.19, 0.26). Significant groundwater level rises were detected in Southern Virginia adjacent to the Chesapeake Bay (0.92 m/yr on average), which could be due to the cessation of groundwater withdrawal from one of the region's largest groundwater users. In the Mid-Atlantic region, shallow groundwater was found to have slight rising trends (0.08 m, p < 0.05) while deeper groundwater showed distinctive declining trends (1.36 m, p < 0.05) between 2002 and 2016. There were significantly more cultivated areas with declining groundwater levels (88% vs. 35%, p < 0.05) in counties with high percentages of cropland (> 50%) compared to areas covered by less cropland. As climate and human pressures increase, it will be critical to identify and evaluate alternative water sources, such as reclaimed water, to sustain agricultural production and protect groundwater resources.