The worm Eudrilus eugeniæ is a species adapted to tropical climates and is highly demanded as fishing bait, therefore possessing high economic value. Separation of worms from humus is typically performed by manual, mechanical, behavioral and electric methods. The objective of this study was to characterize electric pulses for separation of worms (Eudrilus eugeniæ) from humus. To determine separation efficiency of worms from humus, a controlled pulse generator was used with frequencies of 1 and 5 Hz, voltages of 100 and 200 V, peak widths of 2, 3 and 4 x 10-3 s and exposure times of 15 and 30 min. The electrical pulse characteristics which resulted in greatest separation efficiency were: 1 Hz, 2 x 10-3 s and 200 V; and the maximum displacement index was 80% which occurred under these conditions. The survival index was 100% for the pulses which resulted in the greatest separation indices.