The Lower Mississippi River contributes significantly to the biodiversity and ecological stability of the alluvial valley, but agricultural, industrial, and municipal developments have historically impacted environmental quality of the river. Toxicity of sediment and sediment pore water was used to assess the current effects of major cities on sediment quality along the Lower Mississippi River. Composite sediment samples were collected from four sites upriver and four sites downriver of five major cities: Cairo, IL; Memphis, TN; Vicksburg, MS; Baton Rouge, LA; and New Orleans, LA. Acute toxicity was determined by exposing Hyalella azteca to solid-phase sediment for 10 days with two water renewals per day and to sediment pore water under static conditions for 96 h. After the initial tests, animals were exposed to ultraviolet light for 16 h. Sediments were analyzed for organics (organochlorine pesticides, PCBs, organophosphate insecticides, and PAHs) and metals (Cr, Cu, Pb, Mn, Ni, Zn). With the exception of upriver from Memphis, solid-phase sediments were not toxic to H. azteca. Pore water from sediments collected upriver of Memphis also showed slight toxicity. Exposure of H. azteca to ultraviolet light did not increase the toxicity of the sediment or pore-water samples, indicating a lack of toxicity from PAHs that are photoactivated by ultraviolet light. Chemical analyses did not reveal any contaminant levels of concern in the sediments. Based on toxicity testing and chemical analyses, quality of sediments collected from the Lower Mississippi was good, with the exception of sites sampled upriver of Memphis.