giftighet av nanosølv for fisk i norske vann og vassdrag
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Giftighet av nanosølv for fisk I norske vann og vassdrag. Silver - the most common material used in commercial nano -products. Nanosilver from a washing machine. Concentration in effluent: 2,7 µg/L to 25 µg/L Average: 10,522 µg/L Large variability!. Particles found with TEM: - PowerPoint PPT PresentationTRANSCRIPT
Eivind Farmen 118.03.2013
Giftighet av nanosølv for fisk I norske vann og vassdrag
Eivind Farmen 2
Silver - the most common material used in commercial nano-products
18.03.2013
Nanosilver from a washing machineConcentration in effluent: 2,7 µg/L to 25 µg/LAverage: 10,522 µg/LLarge variability!
Particles found with TEM:Size range: 6 – 11 nm
Farkas (2011) Environment international 37 (6), 1057-1062
Objectives• Determine acute and sublethal toxicity of AgNP to fish & fish cells
• Determine AgNP characteristics in various matrixes
• Compare toxic potential of AgNP to ionic silver and AuNP
• Assess risk to fish under ecologicaly relevant exposure scenarios
18.03.2013Eivind Farmen
In vitro experimental modelWhat are the possible effects of nanoparticles on fish cells used as an in vitro test system?
LiverGill
Exposure
(48h)
CytotoxicityReactive oxygen species (ROS) productionEpithelial integrity
Primary hepatocytes
Primary gill epithelium cells
18.03.2013Eivind Farmen
Particle characteristics by TEMAgNPs
Water
cell media
cell media +DOC
Roundish to oval particle
shape
AuNPs
In–house synthesized citrate-coated NPs
ROS production & Cytotoxicity(hepatocytes)Silver NPs Gold NPs
ROS
Cytotoxicity
Farkas et al. 2009. Effects of silver and gold nanoparticles on rainbow trout (Oncorhynchus mykiss) hepatocytes. Aquatic toxicol.
18.03.2013Eivind Farmen
Effects on primary gill cells
Ag is less cytotoxic to gill cells than to hepatocytesAuNPs were not cytotoxic
18.03.2013Eivind Farmen
Eivind Farmen 918.10.2012
What happens in natural water?
Commercial AgNP1:10 in natural lake water
In-house AgNP1:10 in natural lake water
4 nm
220 nm
Eivind Farmen 10
Effects Ag nanoparticles
? ?
Ag in gills
0
10
20
30
40
* **
*
(n=15) (n=15) (n=15)
(n=15)
(n=15)
(n=13)
(n=14)
Ag
(mg/
kg g
ill ti
ssue
)
Control 1 g/LAgNP
20 g/LAgNP
100 g/LAgNP
20 g/LAg+
100 g/LAgNP2
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Eivind Farmen 1118.03.2013
GO analysis AgNP 20 µg/L
Eivind Farmen 1218.10.2012
GO analysis
Eivind Farmen 13
Transcriptional UP-regulation (Ag)100 ug/L AgNP 20 ug/L Ag+
20 ug/L AgNP
Heme-binding protein 2Protein-Glutamine -glutamyltransferase
GTPase IMAP family member 4Na/H exchanger 9
18.03.2013
Eivind Farmen 14
Transcriptional Down-regulation (Ag)100 ug/L AgNP 20 ug/L Ag+
20 ug/L AgNP
Carbonic anhydrase 7Na/K-ATPase
Interferon gammaMHC Class II
18.03.2013
Eivind Farmen 15
Mode of action AgBlood Gill Water
HCO3-
Cl-Cl-
NaK-ATPase
H+ (NH4+)
Na+
K+
Na+
Carbonic anhydrase
CO2 + H2O =>H+ + HCO3
-CO2
Na+
Na
95
105
115
125
135
145
155
*
*
(n=9)
(n=14)(n=14)
(n=9)
(n=4)
(n=10)
(n=13)
m m
ol/L
K+
K
T0
Control
1 µg/L
20 µg
/L
100 µ
g/L
20 µg
/L Ag+
100 µ
g/L NP20
2
4
6
8
10
*
*(n=9)
(n=14)
(n=14)
(n=9)
(n=1)
(n=10) (n=13)
m m
ol/L
Cl-
CO2
Cl
T0
Control
1 µg/L
20 µg
/L
100 µ
g/L
20 µg
/L Ag+
100 µ
g/L NP290
100
110
120
130
140
150
**
(n=9) (n=14) (n=14) (n=9)
(n=1)
(n=10)(n=13)
m m
ol/L
18.03.2013
Eivind Farmen 16
Hazard/risk assessment
100 ug/LAgNP
20 ug/LAg+
20 ug/LAgNP
Control
Toxicity of AgNP due to acting as reservoir for Ag+?
Toxicity of AgNP higher in soft waters !
18.03.2013
Eivind Farmen 17
Effects Ag nanoparticlesAg in gills
0
10
20
30
40
* **
*
(n=15) (n=15) (n=15)
(n=15)
(n=15)
(n=13)
(n=14)
Ag
(mg/
kg g
ill ti
ssue
)
Control 1 g/LAgNP
20 g/LAgNP
100 g/LAgNP
20 g/LAgNP
100 g/LAgNP2
18.03.2013
Osmo-regulation
Necrosis/apoptosis
Eivind Farmen 18
Work in progress
18.03.2013
T0
Control
1 µg/L
20 µg
/L
100 µ
g/L
20 µg
/L Ag+
100 µ
g/L NP20.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
***
*Ag in liver
Ag
(mg/
kg g
ill ti
ssue
)
HSP70
T0
Control
g/L NP1
1
g/L NP1
20
g/L NP1
100
g/L Ag+
20
g/L NP2
100
0.00.51.01.52.02.53.03.54.04.5
Mea
n no
rm e
xpr
T0
Control
g/L NP1
1
g/L NP1
20
g/L NP1
100
g/L Ag+
20
g/L NP2
100
0
1
2
3
4
5
6 *
MT-A
Mea
n no
rm e
xpr
Eivind Farmen 19
Differences Ag-NPs
18.03.2013
GSSG (µM/mg)
T0
Control
g/L NP1
1
g/L NP1
20
g/L NP1
100
g/L Ag+
20
g/L NP2
100
0.00.10.20.30.40.50.60.70.80.9
*µM
GSS
G/m
g liv
er (w
w)
Eivind Farmen 2018.03.2013
Conclusion• High toxicity of Ag-NPs in natural
Norwegian waters• Ag-NP effect mediated by Ag+ ions?• Effects in gills/osmoregulation• Modulation of immune system• Effect in gills > liver
Eivind Farmen 2118.03.2013
Acknowledgements• Farkas J, Mikkelsen HN, Evensen Ø, Heier LS,
Einset J, Salbu B, Rosseland BO, Oughton DH, Tollefsen KE, Paul Christian, Julián Alberto Gallego-Urrea, Norbert Roos, Martin Hassellöv, Kevin V Thomas, Mie Jareid, Hanne Fossnes, Hans-Christian Teien
• Funding:– Norwegian Research Council
» Nanotrace Project» NIVA, SIP project 24024 NEWPOLL
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