The Mississippi Delta, located in northwest Mississippi, is an area dense with industrial-level agriculture sustained by groundwater-dependent irrigation supplied by the Mississippi River Valley Alluvial aquifer (alluvial aquifer). The Delta provides agricultural commodities across the United States and around the world. Observed declines in groundwater altitudes and streamflow contemporaneous with increases in irrigation have raised concerns about future groundwater availability and the effects of groundwater withdrawals on streamflow. To quantify the impacts of groundwater withdrawals on streamflow and increase understanding of groundwater and surface-water interaction, hydrograph-separation techniques were used to estimate baseflow and identify statistical streamflow trends. The analysis was conducted using the U.S. Geological Survey Groundwater Toolbox open-source software and daily hydrologic data provided by a spatially-distributed network of paired groundwater wells and streamgaging sites. This study found that effects of groundwater withdrawals on streamflow were observed as statistically significant reductions in baseflow in areas with substantial groundwater-altitude declines. Hydrograph-separation and trend analyses may be applicable to assess the impacts of groundwater withdrawals in altered environments and streamflow may be used as a proxy for changes in groundwater availability. Characterizing and defining hydrologic relations between groundwater and surface water will help scientists and water-resource managers refine a regional groundwater-flow model that includes the Mississippi Delta that will be used to aid water-resource managers in future decisions concerning the alluvial aquifer.