Meeting Abstract
P3.92 Saturday, Jan. 5 Vitellogenesis induction in male fish exposed to municipal wastewater effluent estrogens TUBERTY, Shea R*; BABYAK, Carol; GAYLORD, Matt; ARNOLD, Justin; CARTER, Bethany; GIBBONS, Leigh Ann; SINGER, Erin; Appalachian State University tubertysr@appstate.edu
We investigated environmental endocrine disruption (EED) resulting from exposure to estrogens from the Boone waste water treatment plant (WWTP) effluent in the S. Fork of the New River. Boone has 15,000 residents, while Appalachian State University (ASU) enrolls nearly 15,000 students. The S. Fork has a flow rate of 6.12 gallons/day and the effluent release comprises 25-40% of the downstream flow. Environmental estrogen exposure is indicated by measuring a common biomarker, the egg yolk protein vitellogenin (Vtg), in male fish. Fish and water samples (1L) were collected from two sites: the WWTP effluent release and upstream at the ASU greenway (reference). Estrogens were separated from water samples by C18 solid phase extraction columns (to concentrate samples 1000 fold) and analyzed by GC-MS and HPLC. The optimized protocol resulted in a lower detection limit of 250 ppb. Various derivatizing agents were explored to enhance the GC-MS detection limits. Fish serum proteins were characterized using SDS-PAGE and analyzed for Vtg presence by Western blotting using antibodies specific to our fish Vtg. Serum samples collected in April 2006 (university in session) indicated sixty percent of male hog suckers, Hypentelium nigricans, were producing Vtg (n=5). Analysis of water samples collected in July (university not in session) indicated that estrogen concentrations were below detection limits. Future work will include collection of monthly water and quarterly fish samples to determine if student estrogens cause temporally increased Vtg production in exposed male fish. Occurrence of EED of fishes with various life history strategies and habitat preferences, as well as spring versus fall spawning fish populations, will be monitored.
