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ASSESSMENT OF IN VITRO ANDROGENIC ACTIVITY IN KRAFT MILL EFFLUENT
Citation:
Lambright, C R., L G. Parks, E. Orlando, L. J. Guillette Jr., G Ankley, AND L. E. Gray Jr. ASSESSMENT OF IN VITRO ANDROGENIC ACTIVITY IN KRAFT MILL EFFLUENT. Presented at Triangle Consortium for Reproductive Biology, RTP, NC, January 27, 2001.
Description:
Detection of In Vitro Androgenic Activity in Feedlot Effluent. Lambright, CS 1 , Guillette, LJ, Jr.2, Gray, LE, Jr.1 , 1USEPA, NHEERL, RTP, NC, 2 University of Florida, Dept. of Zoology, Gainesville FL
Recent studies have shown the presence of androgenic activity in water collected downstream from a pulp and paper mill. The current investigation was designed to determine if there were detectable levels of androgenic activity in water collected down from a feedlot in Nebraska(FLE). This feedlot was selected because of the use of growth promoters in the beef cattle. The active component(s) are excreted and accumulate in nearby water sources. One such promoter is trenbolone. In this study, we investigated the in vitro effects of trenbolone as well as examined water from the FLE and a pristine site. Trenbolone induces luciferase in a stable hAR/MMTV-luciferase cell assay in a dose-response manner with significant responses as low as 0.1 fmole. This response is significantly reduced with the addition of 1 uM hydroxyflutamide (OHF). FLE (40 ml concentrated by C18 solid phase extraction (SPE) eluted with 100% methanol) competed with a synthetic androgen for binding to the human androgen receptor (hAR) at 57.1% of control. Unextracted FLE was also found to induce AR-dependent transcriptional activation in CV-1 cells (cotransfected with pCMVhAR and MMTV luciferase reporter) with 11.1 fold induction (reference site = 1.1 fold and DHT = 18.9 fold). Dilutions of the FLE to 50%, 25% and 10% resulted in a ?dose-dependent' decrease in luciferase expression. The androgenic activity of FLE was reduced by 1 uM OHF. Further studies are ongoing to identify the androgenic component(s) in the FLE.
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