Fish full life-cycle testing for the weak estrogen 4-tert-pentylphenol on medaka (Oryzias latipes)

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Abstract
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 μg/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 μg/L. The LOECs for estrogenic effects (as shown by abnormal sexual differentiation and VTG induction) were 224 and ≤51.1 μg/L, respectively, and the LOEC for reproductive impairment was 224 μg/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 ≤51.1 μg/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.