Abstract:Fish and habitat were sampled by state agencies at 48 stations throughout the Cache River watershed, Illinois between 1992 and 2009. Two distinct fish assemblages were identified, one primarily found in the lower mainstem Cache River and a second found throughout tributaries and the upper mainstem Cache River. Using a canonical correspondence analysis, the distribution of fish species was largely explained by substrate, land use, drainage area and local habitat features. Creek chub, central stoneroller, fringed darter and fantail darter are species found to be positively associated with gravel substrate and forest. In contrast, black buffalo, gizzard shad, smallmouth buffalo, freshwater drum and bigmouth buffalo were positively associated with drainage area, silt, channel width and row crops. Cobble appears to be rare habitat associated with fringed darter, freckled madtom and fantail darter. Results suggest that substrate, land use and local habitat features influence fish assemblage within the Cache River watershed. This information contributes to both understanding aquatic community structure in a highly altered yet diverse watershed as well as management activities within the Cache River watershed.3