Three small fish species, medaka (Oryzias latipes), fathead minnow (Pimephales promelas), and zebrafish (Danio rerio), were exposed to an estrogen, 17 beta-estradiol (E2), and an androgen, 17 beta-trenbolone (TB), for 21 d under flow-through conditions to compare the susceptibility among these three small fish species to the substances. Effects on gross morphology, including secondary sex characteristics and gonadosomatic index, as well as on blood or liver vitellogenin (VTG) levels were assessed. In E2 exposures, significant increases in estrogenic activity were observed in both sexes of all three fish species. The lowest-observed-effect concentrations (LOECs) of E2 for VTG induction in males of medaka, fathead minnow, and zebrafish were less than or equal to 8.94, 28.6, and 85.9 ng/L, respectively. In TB exposures, we observed masculinization of secondary sex characteristics in females as a result of the androgenic activity of TB in medaka with a LOEC of 365 ng/L and in fathead minnow with a LOEC of 401 ng/ L. We also found VTG reduction in females of all three fish species. These results suggest that the susceptibility of medaka to estrogenic chemicals may be higher than those of fathead minnow and zebrafish and that the susceptibility of medaka to androgenic chemicals may be almost equal to that of fathead minnow in the 21-d fish assay.

We studied the chronic effects of 4-nonylphenol (4-NP) on reproductive status of medaka (Oryzias latipes) over two generations of continuous exposure. The exposure study of the parental (F0) medaka was begun on embryos within 24 h postfertilization and continued with monitoring through embryological development, hatching, posthatch survival, growth, sexual differentiation, and reproduction under flow-through exposures to mean measured 4-NP concentrations of 4.2, 8.2, 17.7, 51.5, and 183 microg/L for up to 104 d. Eggs spawned from the F0 fish at 102 and 103 d posthatch were also examined for hatchability, survival after hatching, growth, and sexual differentiation until 60 d posthatch. The 183-microg/L treatment significantly reduced the embryo survival and swim-up success of the F0 fish. The cumulative mortality after swim-up of the F0 fish exposed to 17.7 and 51.5 microg/L were significantly higher than the control mortality. No concentration-related effect of 4-NP was observed on the growth of surviving F0 fish at 60 d posthatch. However, the sex ratio estimated from the appearance of their secondary sex characteristics was skewed toward female in the 51.5-microg/L treatment. Additionally, gonadal histology showed that 20% of the fish in the 17.7-microg/L treatment and 40% in the 51.5-microg/L treatment had testis-ova, indicating that 4-NP affects the gonadal development and survival of medaka at similar concentrations in juveniles. The sex ratio of the F0 fish in the 51.5-microg/L treatment was completely skewed toward female; subsequently, the effects on fecundity and fertility in this generation were monitored at mean measured concentrations of 4.2, 8.2, and 17.7 microg/L from 71 to 103 d posthatch. Fecundity was unaffected by any of the treatments examined. The mean fertility in the 17.7-microg/L treatment was reduced to 76% of that in the controls, although no statistically significant differences were determined. Overall, these results indicate that the lowest-observed-effect concentration (LOEC) and no-observed-effect concentration (NOEC) of 4-NP through the life cycle of the F0 medaka were 17.7 and 8.2 microg/L, respectively. In the F1 medaka, no significant effects were observed on hatching success, posthatch mortality, or growth, but sexual differentiation at 60 d posthatch was affected. Induction of testis-ova in the gonads of the F1 fish was observed in both the 8.2- and the 17.7-microg/L concentrations. The results indicate that 4-NP can have significant effects on reproductive potential of medaka at concentrations as low as 17.7 microg/L.

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.

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