To better understand the effects of hydrologic alteration as they relate to human and biological needs within the Mississippi Alluvial Plain of the south-central United States, the quantification of hydrologic alteration is required. Quantifying hydrologic alteration in the Mississippi Alluvial Plain is particularly difficult because of the lack of current reference, or even relatively undisturbed, U.S. Geological Survey (USGS) streamflow-gaging stations. Water withdrawals for agriculture in the form of weirs, dams, channelization and other forms of regulation within the Mississippi Alluvial Plain increased substantially beginning around 1960 suggesting that streamflow is substantially altered after this time period. To overcome the lack of stations that exist in the present that are unaffected by anthropogenic activities (or current reference stations), historical streamflow data were used to estimate what streamflow would be in the present without anthropogenic influence (or current reference conditions). These data, when combined with current streamflow information collected by the USGS in south and eastern Arkansas, southwest corner Kentucky, Louisiana, western Mississippi, southeastern corner Missouri, extreme western Tennessee, and extreme southeastern Texas were used to assess the level of hydrologic alteration within the study area.