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Characterizing streamflow changes in the agricultural U.S. Midwest is critical for effective planning and management of water resources throughout the region. The objective of this study is to determine if and how baseflow has responded to land alteration and climate changes across the study area during the 50-year study period by exploring hydrologic variations based on long-term stream gage data. This study evaluates monthly contributions to annual baseflow along with possible trends over the 1966-2016 period for 458 U.S. Geological Survey streamflow gages within 12 different Midwestern states. It also examines the influence of climate and land use factors on the observed baseflow trends. Monthly contribution breakdowns demonstrate how the majority of baseflow is discharged into streams during the spring months (March, April, and May) and is overall more substantial throughout the spring (especially in April) and summer (June, July, and August). Baseflow has not remained constant over the study period, and the results of the trend detection from the Mann-Kendall test reveal that baseflows have increased and are the strongest from May to September. This analysis is confirmed by quantile regression, which suggests that for most of the year, the largest changes are detected in the central part of the distribution. Although increasing baseflow trends are widespread throughout the region, decreasing trends are few and limited to Kansas and Nebraska. Further analysis reveals that baseflow changes are being driven by both climate and land use change across the region. Increasing trends in baseflow are linked to increases in precipitation throughout the year and are most prominent during May and June. Changes in agricultural intensity (in terms of harvested corn and soybean acreage) are linked to increasing trends in the central and western Midwest, whereas increasing temperatures may lead to decreasing baseflow trends in spring and summer in northern Wisconsin, Kansas, and Nebraska.