We repeated this for each
tank following its 24-hour exposure with a new euthanasia bath prepared
for each tank. In total, we had four final treatments, each in quadruplicate:
freshwater challenge to freshwater anesthetic, saltwater
challenge to freshwater anesthetic, saltwater challenge to saltwater anesthetic,
and freshwater challenge to saltwater anesthetic. All fish lost
equilibrium in less than 20 s (stage 4 anesthesia) after being placed in
the anesthetic and were unresponsive to external stimuli in less than
90 s (stage 5 anesthesia, as defined