Behavioral analysis
Experiments were carried out at 5-dpf. At 5-dpf, the larvae were
collected, cleaned and placed in 96-well plates. Each well contained 0.2
mL of fish water and one larva. Behavioral tests were performed as
following: the larvae were allowed to acclimate for 15 min before
locomotion monitoring (Zhao et al., 2012). Next, the larvae were allowed
to freely explore the aquarium for 30 minutes. A camera positioned above
the plate was used for movement tracking. All digital tracks were
analyzed by Ethovision XT software (Noldus Information Technology,
Wageningen, Netherlands), and system noise was filtered out with a
minimum movement distance of 0.2 mm. Four parameters, including the
total movement distance, velocity, mobility and maximum acceleration
were analyzed.