Steroid sex hormones,such as estradiol-17β(E2)and testosterone(T),are important regulators of sex change in fish.In this study,we examined the effects of E2 treatment on the dynamics of E2 and T during gonadal differentiation in the olive flounder Paralichthys olivaceus using histology and radioimmunoassay(RIA).Flounder larvae were divided into five groups(G0–G4),and fed with 0 (control),0.2,2,20 and 100 mg E2/kg feed from 35 to 110 day post hatching(dph).Fish growth in the G1 and G2 groups was not significantly different from that of the control group(P>0.05),while fish in the G3 and G4 groups were less active and showed growth depression and high mortality.The gonads of fish in the G3 and G4 groups were smaller and surrounded by hyperplastic connective tissue.The frequency of females in the G0–G4 groups was 54.5%,75.0%,100%,100%and 93.3%,respectively.The RIA analyses of E2 and T showed that T levels decreased during gonadal differentiation,and increased slightly at the onset of ovarian differentiation,while E2 levels increased gradually and peaked at the onset of ovarian differentiation in the control group.In the E2-treated groups,T levels decreased before the onset of ovarian differentiation.E2 levels were high on the 48 dph,but declined to a lower level on the 54 dph, and then increased gradually during gonadal differentiation.And a sharp increase of E2 levels were observed in all E2-treated groups at the onset of ovarian differentiation.The data suggest that T and E2 play important roles during gonadal differentiation,and an E2 dose of 2 mg/kg feed could induce sex reversal in P.olivaceus. Steroid sex hormones, such as estradiol-17β (E2) and testosterone (T), are important regulators of sex change in fish. In this study, we examined the effects of E2 treatment on the dynamics of E2 and T during gonadal differentiation in the olive flounder Paralichthys olivaceus using histology and radioimmunoassay (RIA) .Flounder larvae were divided into five groups (G0-G4), and fed with 0 (control), 0.2, 2, 20 and 100 mg E2 / kg feed from 35 to 110 days Post hatching (dph). Fish growth in the G1 and G2 groups was not significantly different from that of the control group (P> 0.05), while fish in the G3 and G4 groups were less active and showed growth depression and high mortality. gonads of fish in the G3 and G4 groups were smaller and surrounded by hyperplastic connective tissue. The frequency of females in the G0-G4 groups was 54.5%, 75.0%, 100%, 100% and 93.3% respectively. The RIA analyzes E2 and T showed that T levels decreased during gonadal differentiation, and increased slightly at the onset of ovarian differentiation, while E2 levels increased gradually and peaked at the onset of ovarian differentiation in the control group. the E2-treated groups, T levels decreased before the onset of ovarian differentiation. E2 levels were high on the 48 dph, but declined to a lower level on the 54 dph, and then increased gradually during gonadal differentiation. And a sharp increase of E2 levels were observed in all E2-treated groups at the onset of ovarian differentiation. The data suggest that T and E2 play important roles during gonadal differentiation, and an E2 dose of 2 mg / kg feed could induce sex reversal in P. olivaceus.