three controlled temperatures of 23, 26 and 29 °C based on a previous study on the optimal water temperature range for larviculture of A. clarkii [14].
2. Materials and methods
2.1. Fish source
2.1.1. Broodstock
A. clarkii individuals were captured from the tropical Sanya waters, Hainan Province, in August 2007, and reared in pairs (one female and one male) in captivity locally until spawning was observed using natural seawater (25–27 °C, 33–35‰ and pH 8.0–8.2) after pass through a sandy filter.
2.1.2. Experimental larvae
Newly-hatched larvae of A. clarkii for the experiment were collected from the same clutch of fertilized eggs of the same pair.