Lecture 6b, Using Area Fish to Aid in Pollution Source Identification and Assess Exposure to Xenoestrogens, Mercury and other Elements

Transcription

1 Lecture 6b, Using Area Fish to Aid in Pollution Source Identification and Assess Exposure to Xenoestrogens, Mercury and other Elements Course Director: Conrad (Dan) Volz, DrPH, MPH Bridgeside Point 100 Technology Drive Suite 564, BRIDG Pittsburgh, PA office cell Assistant Professor, Environmental and Occupational Health, University of Pittsburgh, Graduate School of Public Health Director-Center for Healthy Environments and Communities Director, Environmental Health Risk Assessment Certificate Program

2 The Basis for Biomonitoring Ecological Receptors as Sentinels for Human Exposure Contaminants in sediments and water bioaccumulate in aquatic food chains and concentration levels can be magnified over 100X in higher trophic level feeders. Lipophilic (hydrophobic) chemicals are more rapidly exchanged between the water and organism than they are excreted or biodegraded by the organism. The channel catfish is a high fat, mid- trophic level, piscivirous fish. Fish classically bioaccumulate lipophilic steroid estrogens and estrogen mimicking chemicals such as estrone (E1), 17-ethinylestradiol (EE2), NP-nonylphenol, DDT and PCB s. These are also stored in human adipose tissue.

3 Channel Catfish estrogenicity is associated with the density of sewer overflows; Implications for population exposure to pharmaceutical estrogens and xenoestrogens.

4 Contraceptives, Female Hormone Replacement Drugs Household products- (Detergents Shampoo etc.) WWTP Aquatic Ecological Receptors Terrestrial Ecological Receptor Humans Runoff Soil and Vadose Zone Surface Water Sedimentation Resuspension Sediment Direct Contact Ingestion Deposition Leaching Direct Contact Ingestion, Food Chain N/A Gills, Dermal, Ingestion Dermal, Ingestion Dermal, Ingestion Dermal, Ingestion Dermal, Ingestion Recharge Outflow Groundwater Direct Contact Ingestion NA NA Dermal, Ingestion

5 Objective A: To determine if the estrogenicity index of channel catfish fillet significantly varies by proximity to dense concentrations of CSOs/ SSOs, as measured by the MCF-7 and BT-20 human breast cancer cell proliferative assays. Objective B: To determine if the channel catfish Estrogen (E)-screen model can be used to help identify the sources of pharmaceutical estrogen and xenoestrogen pollution.

6 Objective C: To involve semi-subsistence and recreational anglers in a Community Based Participatory Environmental Research Project (CBPR). Objective D: To demonstrate to policymakers the known biological effects and potential environmental public health consequences of failure to update Allegheny County s and southwestern Pennsylvania s antiquated sewer systems.

7 Locations of Catfish Catch in Southwestern Pennsylvania Highland Park Point State Park Monongahela River near the U. S. Steel Edger Thompson Works

8 Arc View 9.1 Map of CSO/SSO Locations in the ALCOSAN Sewershed and Channel Catfish Sampling Locations

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10 Number of Channel Catfish Samples Plated on MCF-7 and BT-20 Cell Proliferation Assays by Location Location Channel catfish Braddock Dam (Monongahela River) 6 Highland Park Dam (Allegheny River) 3 Kittanning (Allegheny River) 6 Point 4 Store-bought (farm raised in Georgia) 2 n=21 Methods 1. Catfish were randomly selected from all samples from each location based on a stratification of gender-male, female and un-identifiable. 2. Each catfish flesh+fat sample was homogenized and hormonally active substances extracted with chloroform-methanol solution (80:20,v/v), nitrogen evaporated residues were soluabilized in ethanol-glycerol (3:1, v/v) and serially diluted to 1/100, 1/200, 1/500, 1/1000, 1/1500, 1/2000, 1/3000 and 1/4000 with steroid free medium (includes 5% dextran-charcoal treated serum) wells of a 96 well plate were background absorbance controls (medium and reagent), the remainder of cells on each plate were seeded with either 5000 MCF-7 or BT-20 cells and 5 replicates of each fish dilution, 10 replicates of 1nM E2 (+ control) and 10 replicates of steroid free medium.

11 Estrogenicity Index Estrogenicity Index is a normalized value. EI=Proliferation Index at each dilution/ Mean Estradiol Response per plate.

12 Average Estrogenicity Index of All Fish At Each Location for Each Dilution Average Estrogenicity Index Dilution Braddock Kittanning Point Highland Park Dam Store-bought

13 River Sampling Locations, CSO/SSO Density and Mean, Minimum, Maximum and SD of EI at 1/100 Dilution Sampling Location Point State Park Braddock Dam Highland Park Dam Kittanning Dam River Allegheny/ Monongahela Confluence, Ohio Monongahela Allegheny Allegheny CSOs/SSOs Radius of 1 Mile CSOs/SSOs 1 mile upstream Number Samples of Mean Estrogenicity Index, 1/100 Dilution Min EI Max EI Std. Dev. EI

14 Regression Results Using Estrogenicity Index Coef. Std.Err z p> z 1/ / / / / / / Braddock HPD Store Point cons sigma_u sigma_e rho Likelihood-ratio test of sigma_u=0: chibar2(01)= Prob>=chibar2 = Observations: 840 LR chi2(11) = Number of fish: 21 Prob > chi2 = Observations per fish: 40

15 Wald s Test Pair Wise Comparisons of Proliferation Index Data store HPD Kittanning Braddock HPD p=0.54 Kittannning p=0.34 p=0.75 Braddock p=0.01 p=0.03 p=0.03 Point p<0.01 p=0.01 p=0.01 p=0.49 Estrogenicity Index Data store HPD Kittanning Braddock HPD p=0.63 Kittannning p=0.30 p=0.57 Braddock p=0.01 p=0.03 p=0.05 Point p<0.01 p<0.01 p<0.01 p=0.15

16 Mean Estrogenicity Index by Fish Extract Dilution: Point State Park and Braddock Dam [Dense CSO/SSO] VS Highland Park Dam and Kittanning Dam on the Allegheny River [Less Impacted by CSOs/SSOs] Estrogenicity Index Dilution = 1/ Point and Monongahela Rivers Allegheny River

17 Conclusions Estrogenic activity of catfish flesh and fat does vary significantly by spatial location to dense concentrations of CSOs/SSOs. Non-estrogenic proliferation factors seem not involved as BT-20 cells were not affected. Catfish can serve as biomonitors for estrogenic pollution. The Community-Based Participatory Environmental Research Project energized anglers and river communities to become stewards of the river-this has led to further foundation support for a spring sampling project on the Allegheny River.

18 Public Health Implications River water is the source of tap water for over 1,200,000 in the Pittsburgh region. 16 billion gallons of sewage overflow is deposited in source drinking water for Pittsburgh per year. Pharmaceutical estrogens and xenoestrogens are hypothesized to be released in even higher concentrations from CSOs/SSOs than from WWTP. There is no regulations requiring WWTP or water treatment facilities to test for or treat water for estrogenic substances. Large susceptible population groups may be at risk for estrogenic exposure through municipal drinking water sources.

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20 Mercury Concentration Local Store Bought Median: ppm Low Tukey Post Hoc Comparison 1 P < %CI of median ratio: 3.72~10.2 Pittsburgh Pool Median: ppm Tukey Post Hoc Comparison 2 P < %CI of median ratio: 1.91~4.95 High Kittanning Dam Median: ppm

21 Selenium in Channel Catfish

22 Arsenic in Channel Catfish

23 Risk Assessment, Hg in Kittanning Fish ADD = Cfish * IRfish * EF * ED BW * AT Where: ADD = Average Daily Dose (ingestion of contaminated fish, mg/kg-day) Cfish = Concentration of contaminants in fish (mg/kg fish) IRfish = per capita intake rate of fish (kg fish/meal) EF = exposure frequency (meals/day) ED = exposure duration (days) BW = body weight (kg) and AT = averaging time (days)

24 Hazard Quotient-Hg Effects, noncarcinogenic Subgroup ADD (range, mg/kg-day) 3-8 yr yr Women of childbearing age Adults RfD HQ

25 Acknowledgements The project is funded by the Highmark Foundation, and the DSF Charitable Trust through the University of Pittsburgh Cancer Institute, Center for Environmental Oncology of the UPCI. Major project support was also provided by the Heinz Foundation, Alle-Kiski Health Foundation and the Department of Environmental and Occupational Health, University of Pittsburgh. CDV and DD are supported by the Centers for Disease Control and Prevention s, Environmental Public Health Tracking (EPHT) Program through the University of Pittsburgh Academic Center of Excellence (UPACE) in EPHT. Program partners including Venture Outdoors and Clean Water Action, Rachael Carson Homestead, Alle-Kiski Health Foundation and RiverQuest.