Characterization of the endocrine potencies of municipal effluents - - PowerPoint PPT Presentation

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Characterization of the endocrine potencies of municipal effluents - - PowerPoint PPT Presentation

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School of Environment and Sustainability Characterization of the endocrine potencies of municipal effluents across Canada using in vitro bioassays Tabata Bagatim Supervisor: Dr. Markus Hecker abata Bagatim 1 , Sara Hanson 2 , Hongda Yuan 2 , Kean

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

Characterization of the endocrine potencies of municipal effluents across Canada using in vitro bioassays

Tabata Bagatim Supervisor: Dr. Markus Hecker

T abata Bagatim1, Sara Hanson2, Hongda Yuan2, Kean Steeves2, Steve Wiseman2, Natacha Hogan2,3, Alice Hontela4, Paul Jones1,2, John Giesy2,5, Leslie Bragg6, Hadi Dhiyebi6, Mark R. Servos 6, Charles Gauthier7 ,François Gagné 8, and Markus Hecker1,2

1School of the Environment and Sustainability, University of Saskatchewan, Saskatoon, SK, Canada; 2Toxicology Centre, University of Saskatchewan, Saskatoon, SK,

Canada; 3 Department of Animal and Poultry Science, University of Saskatchewan, Saskatoon, SK, Canada; 4Department of Biological Science, University of Lethbridge, Lethbridge, AB, Canada; 5Department of Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, SK, Canada; 6 Biology Department, University of Waterloo, Watereloo, ON; 7INRS-ETE et UQTR, Quebec, QC, 8 Environment Canada

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

Endocrine Disrupting Compounds

  • There is increasing concern about chemicals with the potential to

adversely affect the endocrine system of humans and wildlife.

  • EDCs of primary toxicological concern:

(anti)estrogenic, (anti)androgenic and steroidogenesis disruption properties.

  • Receptor mediated processes
  • Non-receptor mediated processes

Figure 1 - EDCs mimicking endogenous hormones http://www.precisionnutrition.com/all-about- environmental-toxin

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

Adapted from Kirsten Moore et al. 2011

MWWEs are considered to be the major source of EDCs in Canadian surface waters.

Sources of EDCs

Figure 2 – Sources of EDCs (Adapted from Kirsten Moore et al. 2011)

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Oct 6th, 2015 CEW 2015

School of Environment and Sustainability Plasticizers

  • Bisphenol A
  • Phthalates

Pharmaceuticals

  • Birth control
  • Cimetidine

Flame retardants

  • Organobromide

compounds Life-Stock Operations

  • Hormones
  • Pharmaceuticals

Household Cleaning Products

Emergent Contaminants

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

EDCs in MWWEs and challenges

  • Incomplete understanding of the effluents from WWTPs contribution to the

environment in Canadian surface waters.

  • Complex mixture – problematic to identify the specific compounds that

are responsible for biological effects in exposed organisms.

Figure 3 – Saskatoon WWTP

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

EDCs identification approaches

  • Traditional targeted chemical analyses is not able to provide a complete

and objective exposure assessment.

  • Targeted in vitro bioassays can characterize the specific endocrine

activity of complex mixtures, including unknowns chemicals.

  • In vitro bioassays have the potential to serve as predictors of potential

hazards for wildlife.

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Toxicology Centre

October 6, 2015 CEW

AIME Overall Project

Assessment of Environmental Impacts of Municipal Effluents (AIME)

In Vitro Studies with Validated Bioassays Chemical Analytical Studies In Vivo Studies with Native Species (FHM)

FHM Reproductive Assays Wild Fish Study (In Stream)

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

Determine whether MWWEs represent a significant source of EDCs to aquatic environments in Canada using an in vitro bioassay-directed analysis approach.

  • Evaluate and quantify endocrine disrupting activities of MWWEs across Canada.
  • Characterize the efficiency of WWTPs to remove EDCs.
  • Evaluate different treatment levels of WWTPs (primary, secondary, etc.).
  • Determine whether season (temperature) influences EDCs removal efficiency.

Objectives

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

Saskatoon 260,600 Pop Regina 232,890 Pop Guelph 134,894Pop Kitchener 231,488 Pop Quebec City 321,221 Pop Montreal 1,900,000 Pop

Methods

Figure 4 – Locations of wastewater treatment plants (WWTPs) in Quebec, Ontario and Saskatchewan, and the population (Pop) they are serving.

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

Methods

Figure 5- Particulate Filtration. Figure 6 - SPE – Capture of organic compounds. Figure 7 - SPE – Sample preparation for bioassay.

MDA (Anti) androgenicity MVLN (Anti) estrogenicity H295R Steroidogenesis Disruption

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

Cytotoxicity Test

0.2 0.4 0.6 0.8 1 1.2 Control Regina Saskatoon Guelph Kitchener Montreal Quebec

Fold-Change [SC=1] City (10x concentrated) Control Influent Effluent

* * * * * * *

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

2 4 6 8 10 12 14 16 18 Control 0.1x 0.3x 1x 3x 10x

AEQ [ng DHT] Concentration

Androgenicity

Control Influent Effluent

10 20 30 40 50 60 70 80 90 100 0.1x 0.3x 1x 3x 10x

% Inhibition Concentration

Anti-Androgenicity

Influent Effluent

* * * * * * * * * *

Dose Response - Saskatoon Spring

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

1 10 100

AEQ [ng DHT] City (1x concentrated)

Androgenicity

Control Influent Eflluent

  • 60
  • 40
  • 20

20 40 60 80 100

% Inhibition City (1x concentrated)

Anti-Androgenicity

Influent Eflluent

* * * * * * * * * * * * * * * *

WWTPs across Canada - Spring 2014

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

1 10 100

AEQ [ng DHT] City (1x concentrated)

Summer 2014

Control Influent Eflluent

1 10 100

AEQ [ng DHT] City (1x concentrated)

Spring 2014

Control Influent Eflluent

* * * * * * * * * * *

Androgenicity across WWTPs

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

  • Montreal Effluent (1x) showed androgenic activity (12-14ng/L AEQ) in spring/summer.

0.1 1 10 100 Control 0.1x 0.3x 1x 3x 10X

AEQ [ng DHT] Fold Concentration [x]

Control Inf Sep 14 Eff Sep 14 Inf Sep 17 Eff Sep 17

* * * * * *

Montreal Androgenicity - Summer 2014

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

1 2 3

EEQ [ng E2] City (1x concentrated)

Estrogenicity

Control Influent Eflluent

10 20 30 40 50 60 70 80 90 100

% Inhibition City (1x concentrated)

Anti-Estrogenicity

Influent Eflluent

*

WWTPs across Canada - Spring 2014

  • Montreal Effluent (1x) = 1.78ng/L EEQ in Spring.
  • Segner, 2003 - EE2>1.67ng/L ↓ egg number and fertilization success, delay in time to spawn.
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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

1 1 2 2 3 3 Control Regina Saskatoon Guelph Kitchener Montreal Quebec

Fold-Change [SC=1] City (1x concentrated) Control Influent Effluent

* * * *

Steroidogenesis Disruption across Canada - Summer 2014

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

  • Large number of known and unknown

chemicals are present in effluents

  • So far, identified approx. 400 different

compounds, ranging from pharmaceuticals and personal care products to pesticides

Non-target screening for contaminants in WWTP Effluents

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

Chemical Analytical Data (ng/L)

Endpoint Season Regina Saskatoon Guelph Kitchener Quebec Montreal Atrazine spring <MDL 0.06 0.33 0.61 0.23 10.11 summer 0.08 0.16 0.21 N/A <MDL 1.05 Carbamazepine spring 7.62 0.65 18.90 1.24 15.86 5.33 summer 0.91 8.79 16.55 N/A 2.93 0.54 Clofibrate spring 37.80 39.82 109.35 <MDL 90.77 <MDL summer <MDL 30.93 78.96 N/A <MDL <MDL DEET spring 395.45 10.29 74.38 7.68 168.15 6.98 summer 25.54 3168.79 61.14 N/A 113.70 34.41 Diazepam spring 0.22 <MDL <MDL <MDL 0.48 0.08 summer <MDL 0.11 <MDL N/A <MDL <MDL Ibuprofen spring <MDL <MDL <MDL <MDL <MDL 687.90 summer <MDL <MDL <MDL N/A 1637.31 194.13 Naproxen spring <MDL <MDL <MDL <MDL <MDL 18.73 summer <MDL <MDL <MDL N/A 320.99 <MDL Triclosan spring 28.95 0.23 3.79 1.76 53.36 62.63 summer 0.37 8.74 2.27 N/A 106.08 13.93

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

Summary

  • Most of the WWTPs had a high removal efficiency of androgenic activity.
  • Selected effluents (e.g. Montreal) had significantly increased androgenic

potencies.

  • Removal efficiencies differed significantly among WWTPs
  • Further analysis need to be completed regarding efficiency of different treatment

levels of WWTPs and determine whether population and temperature affects EDCs removal efficiency.

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

Conclusion

  • In vitro assays supported in vivo findings, suggesting a cost-effective tool for

predicting EDCs in aquatic environments.

  • Targeted chemical analysis did not show a presence of chemicals with

estrogenic properties, aligning with in vitro assays findings.

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

Acknowledgement

  • UofS Toxicology Center
  • Saskatoon Wastewater Treatment Plant
  • City of Regina Wastewater Treatment Plant
  • Department of Biological Science, University of Lethbridge, Lethbridge, AB,

Canada;

  • Biology Department, University of Waterloo, Waterloo, ON
  • INRS-ETE et UQTR, Quebec, QC, 8 Environment Canada
  • Ashley Moate, Craig Baird, Shawn Beitel, Leanne Flahr, J.X. Sun, Bryanna

Eisner

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

Questions?

Fig8: FHM field sampling in Saskatoon Upstream and Downstream

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School of Environment and Sustainability

Oct 6th, 2015 CEW 2015

References

1. Wilson et al. 2002 - Toxicol Sci 66: 69-81 2. Wilson et al. 2004 - Toxicol Sci 81: 69-77 3. Hecker et al. 2006 - Toxicol Appl Pharmacol 217: 114-124 4. Hecker et al. 2011 – Environ Sci Pol Res 18: 503-515 5. Mosman et al. 1983 - J Immunol Methods 65: 55-63 6. Hallgren, 2011 - Ecotoxicology 21: 803–810 7. Jobling et al., 1996 – Environ. Toxicol. Chem. 15: 194-202 8. Kime and Nash, 1999 – Aquaculture 177: 345-352 9. Belfroid et al., 1999 - Sci. Total Environ., 225: 101-108 10. Kidd et al., 2014 - Phil. Trans. R. Soc. B 369 11. Segner, 2003 - Ecotox. Environ. Saf. 54: 216–222