Effects on reproductive potential and endocrine status in the mummichog (Fundulus heteroclitus) after exposure to 17alpha-ethynylestradiol in a short-term reproductive bioassay.

A short-term reproductive bioassay with the mummichog (Fundulus heteroclitus) was developed to link changes in endocrine status to reproductive potential subsequent to endocrine disrupting substance (EDS) exposure. Sexually mature mummichog were separated by sex and exposed to the synthetic estrogen 17alpha-ethynylestradiol (EE2) at nominal concentrations of 0-100 ng/L for 21 days using a static daily renewal protocol. Half of the fish were sampled on Day 21. At 100 ng/L, male fish had induction of vitellogenin (VTG), increased gonadosomatic index (GSI), decreased testosterone production and decreased circulating 11-ketotestosterone (11-KT). Female fish had decreased circulating estradiol (E2) and testosterone (T) at 100 ng/L. There were some impacts at lower concentrations of EE2 in both sexes, though the results were not consistent. On Day 21, the remaining male and female fish were combined at each treatment and exposed for an additional 7 days during which spawning and fertilization success were also assessed. Males exposed to 100 ng/L EE2 exhibited VTG induction, increased GSI, and decreased T production on Day 28. Female fish had increased E2 and T production at 1 and/or 10 ng/L and circulating E2 levels remained depressed above 10 ng/L. Female fish exposed to 100 ng/L spawned fewer eggs; fertilization was also impaired. In a parallel exposure, measured EE2 water concentrations were approximately 10-20% of nominal for the 100 ng/L EE2 treatment over a 24-h static exposure; levels in the other treatments were below detectable levels. Fish exposed to nominal concentrations of EE2 below environmentally relevant levels (i.e., <10 ng/L) showed minimal effects while both the endocrine system and reproductive potential were affected at 100 ng/L EE2 (nominal).

Effects of 17alpha-ethynylestradiol on early-life development, sex differentiation and vitellogenin induction in mummichog (Fundulus heteroclitus).

Fertilized mummichog eggs retrieved from 17alpha-ethynylestradiol (EE(2)) exposed adult fish were raised in concentrations of EE(2) ranging from 0 to 100 ng/L (100 ng/L EE(2) estimated to have actual average exposure concentrations of 30% of nominal; 0.1-10 ng/L were below detect throughout 24-h exposure period) for 61 weeks post-hatch. Eggs exposed at 100 ng/L hatched sooner, the larvae were longer, and survival of juvenile fish from hatch to study termination was greater than all other treatments, though fewer hatched at this treatment. Sex ratios were skewed (>80% female phenotype) at 100 ng/L EE(2), and some gonadal male fish displayed female secondary sex characteristics. Condition factor, gonadosomatic index (GSI), and liver somatic index (LSI) were found to decrease in both sexes between 52 and 61 weeks post-hatch. Female fish had increased hepatic vitellogenin (VTG) at 52 weeks post-hatch. When exposed to 1, 10 and 100 ng/L EE(2), female fish had a higher proportion of vitellogenic follicles in the ovarian tissue. Males exposed at 100 ng/L may have had disruption at some endpoints (GSI, VTG) that is masked due to reduced sample size compared to other treatments. Fish exposed to concentrations of EE(2) at or below 10 ng/L showed inconsistent effects on development and reproductive potential. This study indicates the potential for population-level effects at the high range of environmental EE(2) at concentrations equivalent to those at which consistent effects in fecundity in the adult mummichog reproductive test have been measured. This work demonstrates that chronic EE(2) exposure causes developmental effects at concentrations similar to those which cause effects in the shorter-term adult mummichog reproductive test. Effects are at higher concentrations than have been noted for freshwater model species. Whether this is because of species sensitivity or due to differences between freshwater and saltwater availability of EE(2) or its uptake requires further study.