We investigated the effects of air exposure and low temperature on the hematological parameters and gene expressions of heat shock proteins (HSP60, HSP70, and HSP90) in the liver and head kidney of orange-spotted grouper, Epinephelus coioides. Air exposure for 12 minutes resulted in a decrease of alkaline phosphatase activity in plasma, which was not recovered after 24 hours after putting back into water and was significantly different from that in the control group. Air exposure induced an increase in hsp60 gene expression in the head kidney. However, this induction effect stopped after 24 hours back into water and the level of expression was lower than the initial value. In the low temperature experiment, the temperature was reduced from 29C to 20C and then to 14C. The fish were sampled at the initial temperature, 20C, 14C, and one day after the temperature returned to normal. The results showed that low temperature (14C) increased plasma glucose concentration. The gene expression of hsp60 in the head kidney was induced by low temperature and continuously increased, reaching a peak after one day of normal temperature. It is suggested that gene expression of hsp60 could be an indicator for stress.