Download - Oil Sand Process Water (OSPW)
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The in vitro endocrine disruptive effects of untreated and ozone
treated oil sands process water
Y. He, S. B. Wiseman, X. Zhang, M. Hecker, P. D. Jones, M. G. El-Din, J. W. Martin, J. P. Giesy.
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Oil Sand Process Water (OSPW) The extraction process results in the
production of large volumes of wastewater, commonly referred to oil sands process water (OSPW)
o Sands, clay, metals, unrecoverable bitumeno Polycyclic aromatic Hydrocarbons (PAHs)o Naphthenic acids (NAs)
NAs are thought to be a / the key contributor to OSPW toxicity.
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Reclamation Large quantity of OSPW
o Zero discharge policyo Growing volume of stored OSPWo Over 109 m3 stored in various basins
Reclamationo Aging reduces acute toxicity of OSPWo Chronic toxicity remainso Long half-life for natural degradation: ~12 yro Need more aggressive efforts: ozonation ?
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Need to be careful we don’t create new toxic species…. Potentially endocrine disruptors?
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Parent NAs in Commercial Mixture
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78% degradation of NAs
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Production of New Oxidized NAs
NA + O
O3 (20 ppm)
This is good!
This may not be good!
Any low MW NAs formed during ozonation, or hydroxylated NAs resembling steroids, may become more bioavailable and consequently adversely affect aquatic organisms.
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Endocrine Disruption in the Literature
Exposures to tailing pond OSPWo Reduce in vitro production of E2 & T by ovarian and
testicular tissue (slimy sculpin ) (Tetreault et al., 2003)
o Reduce plasma sex hormones in yellow perch (van de Heauvel et al., 1999) & goldfish (Lister et al., 2008)
o Cause longer time to produce first clutch of eggs, and fewer number of clutches of eggs (fathead minnow) (Siwik et al., 2000)
o Reduce plasma T and 11-keto-T in male, and reduce spawning in female fathead minnow (Kavanagh, et al., 2010)
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Objectives To assess the endocrine disruptive
effects of OSPW
To determine the effects of ozonation on the endocrine disruptive properties of OSPW
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Hypothalamus
Pituitary
Testes Ovaries
GnRH
LH
Sex Hormone Production and ActionHPG Axis
Site of Hormone Action
Testosterone Estradiol
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Study 1: Hormone ProductionOzonation attenuates the steroidogenic disruptive effects of sediment free oil sands process water in the H295R cell line
Y. He, S. B. Wiseman, X. Zhang, M. Hecker, P. D. Jones, M. G. El-Din, J. W. Martin, J. P. Giesy. 2010. Ozonation attenuates the endocrine disruptive effects of sediment-free oil sands process water in the H295R cell line. Chemosphere. 80: 578-584.
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The H295R Cell Line
Cholesterol
Pregnenolone
Progesterone
11-Deoxy-Corticosterone
Corticosterone
Aldosterone
17α-OH-Pregnenolone
11-Deoxycortisol
Cortisol
17α-OH-Progesterone
DHEA
Androstenedione
Testosterone Estrone
17β-Estradiol
CYP11A
3β-HSD
CYP21
CYP17
CYP11B2
CYP19
CYP11B2
CYP17
CYP21
CYP11B1
3β-HSDCYP17
CYP17
3β-HSD
17β-HSD
17β-HSD CYP19
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Results: T & E2 production
T production decreased E2 production increased Ozonation attenuates the effects
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Results: Aromatase & E2 metabolism
Aromatase activity increased E2 metabolism decreased Ozonation eliminates the adverse effects at
85%
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Conclusion – Study 1 Ozonation attenuated the adverse effects of
OSPWo T and E2 productiono E2 metabolismo Aromatase enzyme activity
Inhibition of E2 breakdown is the major reason for increased E2 production
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To determine whether OSPW affects androgenicity and estrogenicity through receptor mediated signaling pathways in vitro
To evaluate the effects of ozonation of OSPW on the receptor mediated signaling pathway
Study 2: Receptor signaling
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Background Information Androgen receptor (AR) and Estrogen
receptor (ER)o Play important roles in sexual differentiation of the
reproductive tract, accessory reproductive organs, and other tissues during fetal development
Test systems used are rapid, sensitive, and convenient assays for detecting AR/ER agonist/antagonist :1. MDA-kb2: Transformed with an androgen-
responsive luciferase reporter plasmid2. T47D-kbluc: Transformed with a luciferase reporter
gene containing ERE
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OSPW shows anti-androgenicity and estrogenicity
OSPW shows anti-androgenicity, and ozonation attenuates the effect
OSPW shows estrogenicity, and ozonation does not attenuate the effect
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OSPW is weak AR antagonist
OSPW potentiates the effect of T at lower concentrations..
Anti-androgenicity
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Estrogenicity
Co-exposure with OSPW & E2 shows additive effects on ER activity
Ozonation does not attenuate the effect
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Estrogenicity is receptor mediated
OSPW, ozonated OSPW, and commercial NAs are estrogenic
The estrogenicity act through estrogen receptor-mediated pathway as the effect can be inhibited by a strong antagonist (ICI 182,780)
Ozonation does not attenuate the effect
a, b & c show significant different within groups
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Conclusion – Study 2 Both full-strength and ozonated OSPW can
disrupt receptor mediated androgen and estrogen signaling
o Both full-strength and ozonated OSPW show anti-androgenicity through weak competitor binding
o Both full-strength and ozonated OSPW show estrogenicity
Ozonation attenuates the effect of OSPW on anti-androgenicity, but not on estrogenicity
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OSPW affects sex hormones production (T & E2 ) Ozonation attenuates the adverse effects of OSPWo T and E2 productiono E2 metabolismo Aromatase enzyme activity
OSPW affects receptor mediated signaling (AR & ER )
Ozonation attenuates the effect of OSPW on anti- androgenicity, but not on estrogenicity
Future studies on fathead minnowo Gene expression profileo Circulating hormones levelo Reproduction and embryo development
Summary
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Acknowledgements A research grant from the Alberta Water Research Institute. A Discovery Grant from the Natural Sciences and Engineering Research
Council of Canada. A research grant from Western Economic Diversification Canada, J.P.G. is supported by the Canada Research Chair Program The authors would like to thank Warren Zubot of Syncrude Canada Inc for
supplying the OSPW.