Glucocorticoid Receptor Activity of a Wastewater Effluent-Dominated - - PowerPoint PPT Presentation

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

Liquid Chromatography Tandem Mass Spectrometry

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

Dexamethasone Dose Response

• 20

20 40 60 80 100 1.E-11 1.E-10 1.E-09 1.E-08 1.E-07

Effective Use Percent Concentration (M)

Plate1 Plate2 Plate3 Plate4 Plate5 Plate6 Plate7

GR Activity of the SCR

• 50

50 100 150 200 0.0 10.0 20.0 30.0

DEX-EQ (ng/L) Stream Distance (km)

12-May-14 6-Jun-14 22-Sep-14 1-Dec-14 12-Feb-15

TR AN

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

25 50 75 100 125 150 175 200 0.0 10.0 20.0 30.0

Dex-Eq (ng/L) Stream Distance (km)

February Chemical February Bioassay December Chemical December Bioassay

TR AN

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
• f Public Health
• Thesis Committee: Shane Snyder, David Quanrud, and Jon

Chorover

• Agilent Technologies

References

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