Effects of the synthetic estrogen 17α‐ethinylestradiol on the life‐cycle of the fathead minnow (Pimephales promelas)

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Abstract
A fish full life‐cycle (FFLC) study was conducted for 17α‐ethinylestradiol (EE2) using the fathead minnow, Pimephales promelas. Newly fertilized embryos (2 (0.2, 1.0, 4.0, 16, and 64 ng/L nominal) in continuous flow‐through conditions for 305 d at 25 ± 1°C. Exposure concentrations were verified by 14C‐EE2 radiochemistry, supported by radioimmunoassay, and mean measured values were ≥70% of nominal. For the F0 adult phase until 301 d posthatch, the no‐observed‐effect concentrations (NOECs) for growth, survival, and reproduction (as egg production) were all ≥ 1.0 ng/L. The NOEC values for F1 embryo hatching success and larval survival (at 28 d posthatch) were both ≥ 1.0 ng/L. While statistically detectable changes in F1 growth were evident at 0.2 ng/L, these were not considered to be biologically significant when compared with historical control data. Male fish exposed to EE2 at 4.0 ng/L failed to develop normal secondary sexual characteristics; on the other hand, assumed females exposed to this level of EE2 were able to breed when paired with males that had not been exposed to EE2. Histology of F0 control, 0.2‐, and 1‐ng/L exposed fish at 56 d posthatch indicated an approximate female‐to‐male (F:M) sex ratio of 50:50 (with no ovatestes observed in the control), while fish exposed to EE2 at 4.0 ng/L for 56 d posthatch had a F: M sex ratio of 84:5 (with ovatestes in 11% of fish). After 172 d posthatch, no testicular tissue was observed in any fish exposed to EE2 at 4.0 ng/L. At the same time point, plasma vitellogenin levels were significantly higher in fish exposed to EE2 at 16 ng/L. A lack of sexual differentiation occurred in males at concentrations ≥4.0 ng/L. Taking into account these data, the overall no‐observed‐adverse‐effect concentration was considered to be 1.0 ng/L.