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.
We elucidated the effects of bisphenol A (BPA) on the reproductive capacity (fecundity and fertility) and estrogenic response of adult Japanese medaka (Oryzias latipes) and studied the transgenerational effects (F, generation growth and sex) of this agent on the F1 offspring. Eight mating pairs of medaka were exposed to BPA for three weeks at a measured concentration of 837, 1,720, or 3,120 microg/L. Throughout the exposure period, we observed no reduction of fecundity and fertility in any treatment group and no reduction in survival and growth or any imbalance of the sex ratio of the offspring. However, histologic examination revealed testis-ova in 13% of the males in the 837-microg/L treatment group, 86% of those in the 1,720-microg/L group, and 50% of those that received 3,120 microg BPA/L. In addition, concentrations of hepatic vitellogenin were increased significantly in males treated with BPA at 3,120 microg/L (p = 0.0148). These results suggest that BPA is a weakly estrogenic compound that has the potential to alter sexual development and induce vitellogenin in male medaka but that has no adverse effect on reproductive success.
Abstract-Japanese medaka (Oryzias latipes) were exposed to bisphenol A (BPA) at the sublethal concentrations of 2.28, 13.0, 71.2, 355, and 1,820 g/L (as mean measured concentrations) in the early life stage from fertilized eggs to 60-d posthatch. Except for the growth and sexual differentiation of the fish at 60-d posthatch, no effects were observed on hatching success and time to hatching in embryological stage and on mortality and abnormal behavior and appearance in hatched larvae. The growth of the fish was suppressed with increasing BPA concentrations, resulting in significant differences in both the total length and body weight of medaka at 1,820 g/L compared with the controls. When observed for their external secondary sex characteristics, no males were identified in the 1,820-g/L treatment. In addition, histological examination showed that 32% of fish in the 1,820-g/L group had testis-ova composed of both testicular germ cells and oocytes. Consequently, the lowest effective concentration for the early life stage of medaka was between 355 and 1,820 g/L. Since the environmental concentrations of BPA are usually three orders lower than the lowest effective concentration, BPA alone may not affect the early life stage of wild fish populations.
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