We investigated physiological changes in the freshwater cichlid, fish Maylandia lombardoi in response to different calcium concentrations in rearing water. Four different calcium concentrations (0, 2, 4 and 8 mM) were prepared in rearing water by the addition of ionized calcium (CaO) to examine the effect of various calcium concentrations in rearing water on physiological changes in the cichlid fish, M. lombardoi. Total calcium concentrations in plasma and body increased in a calcium concentration-dependent manner and reached the maximum at 8-mM calcium concentration. Stress-related cortisol was significantly decreased in the 8-mM group compared to the control group (0 mM). Lysozyme activity also significantly decreased in the 8-mM group. These results suggest that CaO in rearing water increases calcium uptake in fish and affects the body by decreasing stress and improving immunity in fish.