We acoustically surveyed relative fish density at the New Cumberland, Hannibal, and Belleville locks, on the Ohio River, using a Simrad EY 500 echo-sounder coupled with a 120 Khz split-beam transducer. The survey was performed prior to the application of rotenone by West Virginia Division of Natural Resources personal; on 15, 16, and 17 September 1998, respectively. Our intent was to strengthen results of acoustic vs. rotenone density comparisons from two sites the previous year. The 1997 surveys on the Ohio River's Willow Island and Racine locks compared favorably with rotenone assessments. Acoustic estimates ranged from -2% to 13% agreement for total fish abundance. Estimates obtained from three 1998 lock surveys provide additional information on which to test the expected 1:1 relationship between acoustic and rotenone estimates.