17beta-estradiol (E2) is a female hormone which is known to be one of the strongest estrogenic chemicals in the environment. The present study investigated the effects of E2 on the reproduction of the estuarine fish, Java-medaka (Oryzias javanicus). Starting from the embryonic stages, Java-medaka was exposed for 6 months to 9.5, 16, 68, 159 and 243 ng/l of E2. The fecundity of Java-medaka exposed to E2 levels >16 ng/l was significantly lower than that of the control. The appearance of secondary sexual characteristics seemed to be inhibited by exposure to 159 and 243 ng/l E2. Vitellogenin concentrations in the liver of male fish exposed to 68, 159 and 243 ng/l were significantly higher than that of the control. Thirty-three and sixty percent of male fishes exposed to 159 and 243 ng/l, respectively, had testis-ova. These results suggest that reproduction in the Java-medaka, especially the male fish, could be affected by exposure to E2 concentrations greater than 16 ng/l.