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IN VITRO ANDROGENIC ACTIVITY OF KRAFT MILL EFFLUENT
Citation:
Parks, L., C Lambright, E. Orlando, L. J. Guillette Jr., G Ankley, AND L E. Gray. IN VITRO ANDROGENIC ACTIVITY OF KRAFT MILL EFFLUENT. Presented at SETAC 21st Annual Meeting, Nashville, TN, Nov. 12-16 ,2000.
Description:
In: Environmental Sciences in the 21st Century: Paradigms, Opportunities, and Challenges: Abstract Book: SETAC 21st Annual Meeting, 12-16 November 2000, Nashville, TN. Society of Environmental Toxicology and Chemistry, Pensacola, FL. Pp. 102.
Female mosquito fish (Gambusia holbrooki) in the Fenholloway River downstream from Kraft mill effluent (KME) outflow dispaly masculinization of the anal fin. The current investigation was designed to determine if KME from this river displayed androgenic activity in vitro. We examined water from 3 sites downstream from the Kraft mill (sites 1,2,3; site 1 is closest to outflow), one upstream (site 4 and another from the Econfina River, FL (site 5), on which there is no Kraft mill. Sites 1-3 (40 ml concentrated byC18 solid phase extraction (SPE) eluted with 100% methanol) inhibited the binding of a synthetic androgen to the human androgen receptor (hAR). Unextracted water from sites 1-3 were also found to display AR-dependent transcriptional activation in CV-1 cells (cotransfected with pCMV hAR and MMTV luciferase reporter) with 15.7 (similar to lnM DHT), 4.6, and 3.8 fold induction. This activity was inhibited by hydroxyflutamide (1microM). Water samples also were separated by eluting fractions from the column using a series of methanol concentrations (25, 50, 75, 80, 85, 90, 95 and 100%) in order to identigy which fraction(s) displayed androgenic activity; these fractions are presently being analyzed.