Glucocorticoid Receptor Activity of a Wastewater Effluent-Dominated Stream Lower Santa Cruz River, Tucson AZ Darcy VanDervort Dr. Shane Snyder Research Group Department of Soil, Water, and Environmental Science College of Agriculture and Life Sciences University of Arizona Special Thanks to: Water Sustainability Graduate Student Fellowship Program Technology and Research Initiative Fund 2014/2015
Wastewater Treatment Plant Discharge 85% of municipal wastewater treatment plants discharge into surface waters in the United States
Glucocorticoid Hormones Human secreted and synthetic Anti-inflammatory properties that treat health problems such as asthma and rheumatic diseases Not filtered out by typical wastewater treatment systems Detected in the environment around the world including US, Netherlands, and China Disrupt biological endocrine systems
Santa Cruz River Sampling Sites
Glucocorticoid Receptor Bioassay Expose genetically modified cells to river extracts Agonists bind to glucocorticoid receptors Quantify by fluorescence
Glucocorticoid Chemical Analysis 28 Target Analytes Aldosterone Flumethasone Amcinonide Flunisolide Beclomethasone Fluocinolone Acetonide Beclomethasone Dipropionate Fluocinonide Betamethasone Fluorometholone Budesonide Fluticasone Propionate Clobetasol Propionate Hydrocortisone Clobetasone Butyrate 6-α-Methylprednisolone Corticosterone Mometasone Furoate Cortisone Prednisolone Deflazacort Prednisone Deoxycorticosterone Acetate Spironolactone Dexamethasone Triamcinolone Fludrocortisone Acetate Triamcinolone Acetonide 8 Surrogates Cortisone-d8 Fludrocortisone-d5 Cortisone-d8 Methylprednisolone-d2 Hydrocortisone-d4 Prednisolone-d6 Dexamethasone-d4 Prednisone-d4 Solid Phase Extraction Liquid Chromatography Tandem Mass Spectrometry
Dexamethasone Dose Response Effective Use Percent 100 80 60 40 20 0 1.E-11 1.E-10 1.E-09 1.E-08 1.E-07-20 Concentration (M) Plate1 Plate2 Plate3 Plate4 Plate5 Plate6 Plate7
GR Activity of the SCR 200 AN TR DEX-EQ (ng/l) 150 100 50 0 0.0 10.0 20.0 30.0-50 Stream Distance (km) 12-May-14 6-Jun-14 22-Sep-14 1-Dec-14 12-Feb-15
Chemical Glucocorticoid Concentrations (ng/l) Sample Distance (km) Cort. Hydrocort. Prednsl. Beta. Triam. Acet. Fluoc. Acet. 12/1/2014 SCR1 0.0 0.52 2.08 0.75 0.47 37.03 1.30 SCR2 1.3 0.39 1.62 0.64 0.54 33.25 1.43 SCR4 7.6 0.21 0.59 0.84 0.22 37.66 1.63 SCR5 9.7 0.19 0.65 0.86 0.26 34.62 1.50 SCR6 13.0 0.16 0.63 0.65 0.35 26.64 1.21 SCR8 17.0 0.27 0.66 0.85 0.43 29.51 1.24 SCR9 24.0 0.11 0.38 0.65 0.33 15.46 0.78 SCR10 28.8 0.11 0.34 0.54 0.24 7.29 0.57 2/12/2015 SCR1 0.0 0.69 1.94 0.97 0.29 28.91 1.42 SCR2 1.3 0.43 1.74 0.59 0.42 22.52 1.00 SCR3 6.4 0.42 1.38 0.48 0.29 8.64 0.53 SCR4 7.6 0.27 0.73 1.06 0.16 31.22 1.22 SCR5 9.7 0.25 0.84 0.94 0.20 26.93 1.09 SCR6 13.0 0.20 0.73 0.84 0.22 18.51 0.93 SCR8 17.0 < 0.04 0.74 0.84 0.30 19.94 0.89
Chemical vs Bioassay Data Dex-Eq (ng/l) 200 AN TR 175 150 125 100 75 50 25 0 0.0 10.0 20.0 30.0 Stream Distance (km) February Chemical February Bioassay December Chemical December Bioassay
Conclusions Decrease in Glucocorticoid concentrations downstream from treatment plants Photolysis Biodegradation Deposition/Sediment sorption Chemical data can t predict GR response Unknown compounds causing response Synergy effects of mixtures
Acknowledgements Shane Snyder Lab Bioassay Team: Ai Jia, Kevin Daniels, Hao Vo Shimin Wu Fred Leusch, Griffith University School of Environment Jeff Bliznick, University of Arizona Soil Water and Environmental Science Matt Kaplan and the University of Arizona Functional Genomics Core Lab Team Kelly Reynolds and Jonathan Sexton, University of Arizona School of Public Health Thesis Committee: Shane Snyder, David Quanrud, and Jon Chorover Agilent Technologies
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