Mature medaka (Oryzias latipes) were exposed to ethinylestradiol (EE2) at measured concentrations of 32.6, 63.9, 116, 261, and 488 ng/L for 21 d under flow-through conditions. Effects on reproductive success of the fish as well as on gonadal condition and vitellogenin (VTG) induction were assessed. A significant decrease in fecundity was observed only at the highest EE2 concentration, whereas hepatic VTG concentration in males was increased at concentrations of 63.9 ng/L and greater. In addition, an intersex condition (testis-ova) of the gonad was observed in male medaka exposed to EE2 concentrations of > or = 63.9 ng/L. Overall, these results indicate that the lowest-observed-effect concentrations of EE2 based on reproduction versus induction of both VTG and testis-ova in the medaka were 488 and 63.9 ng/L, respectively. Thus, the physiological and histological measurements were approximately eightfold more responsive to the EE2 exposure than were overt reproductive effects. This suggests that the elevated VTG levels and testis-ova may not be definitely responsible for reproductive impairment of the fish.

Abstract-We studied the chronic effects of methyltestosterone (MT) on reproductive status of medaka (Oryzias latipes) over two generations under continuous exposure to verify the applicability of the fish full life-cycle test (FFLC) for this androgen with this species. The exposure of parental (F0) medaka to MT was begun on embryos within 12 h postfertilization and continued for up to 101 d; assessment endpoints included embryological development, hatching, posthatch survival, growth, sexual differentiation, reproduction, and hepatic vitellogenin (VTG) levels under flow-through exposure to MT at each mean measured concentration of 0.35, 1.09, 3.29, 9.98, and 27.75 ng/L. Eggs (F1) spawned from the F0 fish at 98, 99, and 100 d posthatch were examined for hatchability, survival after hatching, growth, sexual differentiation, and hepatic VTG level until 60 d posthatch. In the FFLC with medaka, MT induced masculinization of both secondary sex characteristics and gonads. We observed that all F0 fish in the 27.75-ng/L treatment group showed male secondary sex characteristics in which no fish with ovary could be discerned. Several fish with ovaries in F0 and F1 generations treated with 9.98 ng/L showed male secondary sex characteristics. We also observed swollen abdomens in the F0 and F1 female fish in the 9.98-ng/L treatment group. These swollen abdomens were induced by enlarged ovaries and were accompanied with declined fecundity and fertility in the F0 generation. These results indicate that MT reduces the reproductive potential of medaka and that the FFLC with this species is applicable to the evaluation of androgens.

We studied the chronic effects of 4-tert-pentylphenol (4-PP) on reproductive status of medaka (Oryzias latipes) over two generations under continuous exposure, with the goal of verifying the applicability of the fish full life-cycle test (FFLC) for this weak estrogen with this species. The exposure of parental (F0) medaka to 4-PP was begun on embryos within 12 h after fertilization and continued for up to 101 d, with monitoring of embryological development, hatching, posthatch survival, growth, sexual differentiation, reproduction, and hepatic vitellogenin (VTG) levels under flow-through exposure to 4-PP at mean measured concentrations of 51.1, 100, 224, 402, and 931 microg/L. Eggs (F1) spawned from the F0 fish at 99, 100, and 101 d after hatch also were examined for hatchability, survival after hatching, growth, sexual differentiation, and hepatic VTG levels, until 61 d after hatch. In the FFLC with the F0 medaka, the lowest-observed-effect concentration (LOEC) of 4-PP for lethal and sublethal toxicity (as shown by growth inhibition) was 931 microg/L. The LOECs for estrogenic effects (as shown by abnormal sexual differentiation and VTG induction) were 224 and < or = 51.1 microg/L, respectively, and the LOEC for reproductive impairment was 224 microg/L. Therefore, the effective concentrations of 4-PP for abnormal sexual differentiation and reproductive impairment were about four times lower than those for lethal and sublethal toxicity. In the F1 medaka, the LOECs for sublethal toxicity and estrogenic effects were 224 and < or = 51.1 microg/L, respectively. This finding suggests that the continuous exposure to 4-PP over two generations induced these adverse effects at lower concentrations in the F1 generation than those in the F0 generation. Thus, 4-PP has estrogenic effects that reduce the reproductive potential of medaka. The results indicate that the FFLC with medaka is applicable to the evaluation of weak estrogens.

Mature medaka (Oryzias latipes) were exposed to ethinylestradiol (EE2) at measured concentrations of 32.6, 63.9, 116, 261, and 488 ng/L for 21 d under flow-through conditions. Effects on reproductive success of the fish as well as on gonadal condition and vitellogenin (VTG) induction were assessed. A significant decrease in fecundity was observed only at the highest EE2 concentration, whereas hepatic VTG concentration in males was increased at concentrations of 63.9 ng/L and greater. In addition, an intersex condition (testis-ova) of the gonad was observed in male medaka exposed to EE2 concentrations of > or = 63.9 ng/L. Overall, these results indicate that the lowest-observed-effect concentrations of EE2 based on reproduction versus induction of both VTG and testis-ova in the medaka were 488 and 63.9 ng/L, respectively. Thus, the physiological and histological measurements were approximately eightfold more responsive to the EE2 exposure than were overt reproductive effects. This suggests that the elevated VTG levels and testis-ova may not be definitely responsible for reproductive impairment of the fish.