FIGURE 1.

Measuring stereotypical movement of Lumbriculus variegatus. (A) L. variegatus are plated in 6‐well plates 18–24 h before the experiment begins. L. variegatus are alternately stimulated with 20–200 μl pipette tip at the (B) anterior region (shown in gray), to stimulate body reversal, or (C) posterior region (shown in black), to stimulate helical swimming. Worms are stimulated for a total of 5 times/end, with a 5–10‐s interval between stimuli. (D) These movements are objectively scored and recorded as 1 = no movement, 2 = incomplete stereotypical movement, 3 = full stereotypical movement, as previously described by Drewes.¹³ (A–D) is repeated for each L. variegatus before exposure to drug compounds to give the baseline ability to perform these movements. L. variegatus are then tested again 10 min after incubation with drugs and 10 min and 24 h in artificial pondwater only. Data are expressed as a ratio of the movement score after exposure relative to the baseline movement score