Reproductive impairment and induction of alkaline-labile phosphate, a biomarker of estrogen exposure, in fathead minnows ( Pimephales promelas) exposed to waterborne 17 β-estradiol

Abstract

The objectives of this research were: (1) to assess the effects of waterborne 17 β-estradiol [E2; (17 β)-estra-1,3,5(10)-triene-3,17-diol; CAS RN 50-28-2] on the reproduction of fathead minnows ( Pimephales promelas) as a benchmark to which xeno-estrogens can be compared, and (2) to correlate the effects on reproductive function with plasma vitellogenin expression, measured as alkaline-labile phosphorous. Histopathological changes were also noted but are reported elsewhere. Duplicate groups of six fish (3 male and 3 female) were exposed to waterborne E2 at nominal concentrations of 10, 1, and 0.1 nM (2724, 272.4, and 27.24 ng l −1) administered via a flow-through proportional diluter apparatus for 19 days. An ethanol carrier solvent was used at a final tank concentration of 1 ppm v/v in the treated tanks and in the solvent control tanks; the latter did not receive E2. Duplicate control tanks received neither ethanol nor E2. Dissolved E2 concentrations, measured throughout the exposure period using an ELISA, averaged 79% of nominal concentrations in the treated tanks. ELISA-detectable concentrations of E2 were found in all tanks (ranging from 3.5 to 15 ng E2 l −1), including the control and solvent control tanks, which indicated that fish in the untreated tanks may have been the source of some E2. The EC50 (concentration expected to cause 50% effect), based on measured E2 concentrations, for inhibition of egg production was 120 ng E2 l −1 (log 10 EC50=2.08±1.22, ±S.E.). The EC50 for induction of vitellogenin (measured as plasma alkaline-labile phosphate) in males was 251 ng E2 l −1 (log 10 EC50=2.40±0.33, ±S.E.). No vitellogenin induction plateau was observed in females, therefore no EC50 could be calculated. Egg production, expressed as eggs laid per female, was significantly correlated with plasma vitellogenin in both males (linear r 2=0.46, P<0.03) and females (linear r 2=0.81, P<0.0004), though the relationship was stronger with female plasma vitellogenin expression than with males. The primary effect of E2 exposure on female fathead minnows appeared to be alteration of the timing of recrudescence including vitellogenin production. Spawning was inhibited in a way that indicated that exposure to waterborne E2 may have `reset' the cycle of recrudescence toward the beginning of the oogenic cycle. Vitellogenin induction in male fathead minnows was strongly correlated with E2 exposure, but less so with egg production. The results of this experiment link a biochemical indicator of waterborne estrogen exposure, vitellogenin, with a reproductive performance indicator, egg production, an important parameter affecting fish populations in the environment.