Once all three baseline trials were completed, chemical exposure
trials began. The setup of the experimental trials is identical to that
of the baseline, however, 20 μL of fluoxetine solution was administered
via pipette into the tank water 5 h prior to the start of testing.
After 5 h, males were presented with the dummy female and
dummy male simultaneously as described above. As with the baseline
trials, males received 3 trials of chemical exposure. At the end of each
day, males received a 30% water change with untreated water. Fluoxetine
has been found to have a half-life of 64–72 h under similar testing
conditions [13]. Thus, the amount of fluoxetine remaining in the
water was minimal (8–23 ng L−1). However, fluoxetine may remain
present in the tissue of exposed individuals as norfluoxetine, which
may prolong the exposure period of individuals. All baseline and fluoxetine
trials were recorded for later analysis. Males received one
trial every other day. There was also a 48 h period between the last
baseline trial and first fluoxetine trial.