The objectives of this study were to investigate the potential roles of cortisol and gonadal steroids in the phagocytic activity of peripheral blood leukocytes in two teleosts, tilapia (Oreochromis niloticus x O. aureus) and common carp (Cyprinus carpio). An in vitro microtiter plate assay, measuring incorporation of FITC-latex beads into peripheral blood leukocytes, was developed for the first time in teleosts. Peripheral blood leukocytes were cultured in AL medium with tested compounds in a microfluor black plate at 25 degrees C. FITC-latex beads were further incubated for phagocytosis and engulfed fluorescent intensity in phagocytes was detected fluorometrically. Cortisol suppressed leukocyte phagocytosis in a dose (10(-14) to 10(-4) M)- and time (0.5 to 8 h)-dependent manner in tilapia. The glucocorticoid agonist dexamethasone had a suppressive effect similar to that of cortisol, while cortisone and the mineralocorticoid aldosterone had only a weak effect in tilapia. High doses of estradiol and ethynylestradiol, but not of estrone, suppressed phagocytosis in tilapia. No suppressive effect on phagocytosis was observed with various concentrations of progesterone, testosterone, and 11-ketotestosterone. Triiodothyronine was also inactive on phagocytosis. A combination of estradiol and cortisol potentiatingly suppressed phagocytosis. Actinomycin D and cycloheximide blocked the suppressive effects of cortisol and estradiol. Cortisol had weaker suppressive effects on the phagocytosis of leukocytes in common carp than tilapia. Other steroids had no suppressive action on phagocytosis in common carp. It is concluded that the suppressive effects of cortisol and estradiol on phagocytosis in tilapia are mediated by specific glucocorticoid receptors and estrogen receptors, respectively. Cortisol would play a main and important role on the down-regulation of phagocytic activity. Sexual steroids, such as estradiol, also could interact with cortisol to further suppress immunity in tilapia. Differential responsiveness of the immune system to suppressive effects of steroids, among teleosts species, has been demonstrated.