The hydrologic regime of the Illinois River has been altered over the past 100 years. Locks and dams regulate water surface elevations and flow, enabling commercial navigation to continue year round. This study relates changes in water surface elevation to fish abundance in the river, and establishes target criteria for operating locks and dams. Using long-term records of daily river stage, we identified ecologically meaningful hydrological parameters for eight gage locations along the Illinois River. Inter-annual variability of a long-term fisheries dataset beginning in 1957 was related to variability in stage, flood and recession duration, frequency, timing, and rate of change of water levels. Reversals in water surface elevation, maximum stage levels, and length of the spring flood were the most important parameters influencing abundance of age-zero fishes in annual collections. Smallmouth buffalo (Ictiobus bubalus), black crappie (Pomoxis nigromaculatus), freshwater drum (Aplodinotus grunneins), and white bass (Morone chrysops) were most abundant in samples during years that approximated the natural water level regime. Of the 33 hydrologic parameters evaluated for the entire water year from an Illinois River gage site on La Grange Reach, all except average stage in January and Julian date (JD) of maximum stage had moderate or high hydrologic alteration based on the historical range of variation (RVA). The highest degree of hydrologic alteration was for minimum stage levels (1-day, 3-day, and 7-day), rate-of-rise, and rate-of-fall. Other parameters that have been severely altered were 30-day minimum stage, 90-day maximum stage, and the annual number of water level reversals. Operations of the La Grange and Peoria locks and dams could be modified so water level variability would approximate that of the late 1800s, when fish and wildlife resources were abundant. The water regime could be regulated to maintain navigation and improve conditions for native plants and animals without increasing flood damages.