The experiments described in this paper relate known target configurations under controlled conditions to acoustic characteristics of multiple moving fish. We wanted to further our understanding of the interactions between targets and the effects these interactions have on the measurement of the number of salmon migrating in rivers. Multiple targets in various configurations were passed through a horizontally oriented 4° × 10° beam from a split-beam echo sounder. The effects on measurements of target strength, detection probability and target location in the beam are presented. The observed target strength was not dependent on target velocity. There was a reduction in target detection due to the single-target selection criteria implemented by the hydroacoustic system. We mimicked the conditions in a river where a close range fish target may modify the beam geometry allowing detection of previously undetected targets. We demonstrated some of the effects resulting from moving targets into radial alignment and we demonstrated shadowing conditions that can cause extinction of target echoes.