oil sands processes affected water …
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Oil Sands Processes Affected Water …. Steve Wiseman Toxicology Centre University of Saskatchewan. Problem: How To Deal with the Large Volumes of OSPW?. Surface mining industry produces large volumes of OSPW OSPW must be reclaimed as viable aquatic habitat 27 end-pit lakes planned - PowerPoint PPT PresentationTRANSCRIPT
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Oil Sands Processes Affected Water …
Steve WisemanToxicology Centre
University of Saskatchewan
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Surface mining industry produces large volumes of OSPW
OSPW must be reclaimed as viable aquatic habitat 27 end-pit lakes planned First (Base mine lake) will be filled this year with OSPW from WIP (Syncrude) EPLs will eventually flow into the natural system
Problem: How To Deal with the Large Volumes of OSPW?
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Toxicity of 1st EPLs are predicted to persist until 2070
Need to accelerate the detoxification process
Can We Accelerate Detoxification of OSPW ?
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Ozonation Decreases NAs in OSPW?
7 8 9 10
11
12
13
14
15
16
17
18
19
20
21
22
0
2
4
6
8
024681012
Carbon number
Rel
ativ
e In
tens
ity
Rings
O3
OSPW NAs Before Ozonation OSPW After Ozonation with 80 mg O3/L
7 8 9 10
11
12
13
14
15
16
17
18
19
20
21
22
0
2
4
6
8
024681012
Carbon numberR
elat
ive
Inte
nsity
Rings
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Attenuation of EmbryotoxicityFa
thea
d m
inno
w e
mbr
yo s
urvi
val
(168
hpf
)
0
20
40
60
80
100
120
Survival
a a a
b
a a a
b
Survival
Perc
ent e
mbr
yos w
ith p
eric
ardi
al e
dem
a (1
68 h
pf)
0
20
40
60
80
ND
Pericardial Edema
bb
a
c
bb
a
cPericardial Edema
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Attenuation of Developmental Toxicity
Freshwater OSPW O3-OSPW
Mean
Wet
Weig
ht
0.000
0.002
0.004
0.006
0.008
0.010
0.012A
A
B
Freshwater OSPW O3-OSPW
Perce
nt A
dult
Emer
genc
e
0
20
40
60
80
100 A A
B
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No Effect on Estrogenicity of OSPW – in vitro
Estro
geni
c re
spon
se
MediaOSPW
OSPW + ICIO3-OSPW
O3-OSPW + ICI0.0
0.5
1.0
1.5
2.0
2.5
3.0
a a a
bb Ozonation neither attenuated nor potentiated estrogenicity of OSPW.
Chemical(s) in OSPW and ozonated OSPW bind to the ER.
Rowland et al., 2011
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VTG CHG L CHG H
mRNA
Abu
ndan
ce(R
elativ
e to
Fres
hwat
er)
0.0
0.5
1.0
1.5
2.0Egg Envelope Proteins - Females
aa
a
b b b b b b
Could explain the decreased fecundity in female minnows (Kavanagh et al., 2011) and less prominent male secondary sexual characteristics in male minnows.
Egg Envelope Proteins- Males
VTG CHG-L CHG-HmR
NA A
bund
ance
(Rela
tive t
o Fr
eshw
ater
)
0
2
4
6
8FreshwaterOSPWO3-OSPW
aa
b
aa
a a
b
b
No Effect on Estrogenicity of OSPW – in vivo
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Need: Determine the Critical Mechanism(s) of Toxicity of Oil Sands ProcessAffected Water
If we know critical mechanisms of action then we can design assays to monitorfor exposure to OSPW
Problem: How do we monitor for exposure to and effects of OSPW?
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Mechanism(s) of Toxicity of OSPW Because NAs are surfactants, OSPW might have toxicity via narcosis.
CLControl CS NA
OSPW O3-OSPW AC-OSPW
Control CLCS (1hr)
CS (2hr)OSPW
O3-OSPWAC-OSPW NA
05
1015202530
Membrane cholesterol
ug c
hol/m
illio
n ce
lls
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TranscriptomicsGiven the complexity of OSPW there might be multiple mechanisms of toxicity.
Adverse outcome pathways• Processes that lead to toxicity are often initiated at the molecular level
Chemical Direct interaction with receptor
Molecular event(ie. transcriptional response)
+
Cellular processes
Organism level effect
Population level effect
Quantify abundances of transcripts in the livers of male fathead minnows exposedto OSPW might provide some insight into potential mechanisms of toxicity.
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Results 1 : Global Gene Expression
UP(109)
Down(95)
Freshwater -vs- Untreated OSPW
Functional annotation using GO terms and KEGG mapping to identify process indicative of effects of OSPW.
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BiotransformationTranscript Fold ChangeCYP1A 2.1CYP2k19 11.3CYP2k6 10.1CYP2N 2.7CYP2AD2 2.2UGT 5B4 6.3UGT 5F1 -4.3Sulfotransferase 1,3 1.8GST (mitochondrial) 4.5GST (cytosolic) >23.3MDR-2 3.3Aldehyde oxidase 1 3.1Aldehyde dehydrogenase
3.6
Monoamine oxidase 3.2Epoxide hydrolase 2.0
Phase I
Phase II
Phase III
Oxidative metabolism
AhR PXRCAR
OSPW-OC
CYP1AGSTMDRUGT
CYP2 CYP3GSTMDRST
Are organics in OSPW beingmetabolized?
Effect on toxicity?
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Transcript Fold Change
Glutathione synthase 3.1Glutathione reductase 3.2Glutathione peroxidase 1.7Transketolase 2.46-phosphogluconate dehydrogenase 10.1Glucose-6-phosphate dehydrogenase 2.7Nuclear factor like 2 1.8
Oxidative Stress - I
Glutathione metabolism
Pentose-phosphate shunt
Glutathione
Reductase
GSH
GSSG
GSH Synthase
Glutathione Peroxidase
NADP
NADPH
G-6-PDH6-PGDH
Transketolase
H202
H20 + 02
NRF2
AO MOA AlDH EH
GST UGT MDR
ROS
Transcription factor
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Oxidative Stress - IITranscript Fold
ChangeNADH dehydrogenase 1 beta subcomplex subunit 1
1.8
Acyl carrier (mitochondrial precursor)
1.5
Cytochrome b-c1 complex subunit 9
1.5
Cytochrome b561 domain 2 3.3Cytochrome b5a 8.8
Complex I
Complex III
Complex I and III are major sites of productionof ROS
http://en.wikipedia.org/wiki/File:Mitochondrial_electron_transport_chain%E2%80%94Etc4.svg
ROS
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ApoptosisTranscript Fold ChangeApoptosis-inducing factor 3 4.3Apoptosis-inducing factor mitochondrial associated-2
4.1
Poly [ADP-ribose] polymerase 4.8Programmed cell death 4a 1.5DNA damage-regulated autophagy modulator protein 2
> 23.3
Cathepsin b 1.5BCL2/adenovirus E1B 19 kDa protein-interacting protein 3
-1.8
Forkhead box transcription factor O3A -3.3AIF
PARP
Cathepsin b
AIF AIF
ROS
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AhR PXR
CAR
OSPW-OC
CYP1ACYP2KCYP2ADCYP2N
OSPW-OC
nucleus
nrf2AOMOAAlDHEH ROS
Complex IComplex III
Mechanism of Toxicity
GSTUGTMDR
Apoptosis
GSTUGTMDR
mitochondria
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Hemorrhage Pericardial edema Malformation of spine
Effects are similar to those caused by dioxins and dioxin-like chemicals (PAHs).
Development of Deformities
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Biotransformation Enzymes
No change in transcript abundance of cyp1a• No activation of Aryl-hydrocarbon Receptor (AhR)
signaling• No PAHs in OSPW.
Greater transcript abundance of cyp3a • Activation of Pregnane-X-Receptor (PXR)
Ozonation & activated charcoal treatment attenuated the effects
cyp1a cyp3a
Fold-
chan
ge in
abun
danc
e o
f tra
nscri
pt
0.0
0.5
1.0
1.5
2.0
2.5
3.0 Freshwater Untreated OSPW03-OSPWAC-OSPW
aa
a
b
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Oxidative stressReactive oxygen species
Greater conc. of ROS Greater transcript abundance of gst & sod Ozonation & activated charcoal treatment attenuated the
effects
Oxidative stress responsive genes
aaa
baa
a
b
aa
a
b
Control OSPWO3-OSPW
AC-OSPW
Conc
entra
tion
of R
OS
0.00.20.40.60.81.01.21.41.61.82.0
gst sod catFo
ld-ch
ange
in ab
unda
nce
of tr
ansc
ript
0
1
2
3
4
5Freshwater Untreated OSPW03-OSPWAC-OSPW
aaa
b
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Apoptosis
Caspase-activated apoptotic cell death induced by oxidative stress
Ozonation & activated charcoal treatment attenuated the effects
Fold-
chan
ge in
abun
danc
eof
tran
scrip
t
0
1
2
3
4
5 Freshwater Untreated OSPW03-OSPWAC-OSPW
aa
a
b
b
b
a a
aaaa
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The toxicity of OSPW due to dissolved organic compounds.
The mechanisms of the effects of the dissolved organic compounds are unknown.
The identities of the compounds that cause effects are unknown.
Conclusion
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OSPW
Characterization of Chemical Content
Characterization of Biological Effects
Toxicity(250,000
chemicals?)
Development of Appropriate Bioassays
Effects
BiomarkersChemical and
biological characterization of OSPW will lead to strategies to deal
with OSPW
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