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Aquatic ecotoxicity testing of nanoplastics
lessons learned from nanoecotoxicology
Hartmann, Nanna B.; Nolte, Tom; A. Sørensen, Mads; R. Jensen, Pernille; Baun, Anders
Publication date:
2015
Document Version
Publisher final version (usually the publisher pdf)
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Citation (APA):
Hartmann, N. I. B., Nolte, T., A. Sørensen, M., R. Jensen, P., & Baun, A. (2015). Aquatic ecotoxicity testing of
nanoplastics: lessons learned from nanoecotoxicology Kgs. Lyngby: DTU Environment. [Sound/Visual
production (digital)]. ASLO Aquatic Sciences Meeting 2015, Granada, Spain, 22/02/2015
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Aquatic Ecotoxicity Testing of
Nanoplastics
Lessons learned from nanoecotoxicology
Nanna B. Hartmann, Tom Nolte, Mads A.
Sørensen, Pernille R. Jensen, Anders Baun
2015 Aquatic Sciences Meeting
27th February 2015, Granada, Spain
The group
• Professor Anders Baun
• Associate professor Steffen Foss Hansen
• Senior Researcher
Hans-Christian Holten Lützhøft
• Postdoc Nanna Bloch Hartmann
• PhD students:
–
–
–
–
–
–
–
–
Sara Nørgaard Sørensen
Denisa Cupi
Lars Michael Skjolding
Biase Liguri
Katrine Nielsen
Laura Heggelund
Aiga Mackevica
Rune Hjort
What do we do?
Hazard
Identification
NanoREM
NanoDen
MARINA
EnvNano
New paradigm
for ecotoxicity
testing of NMs
(European
Research Council
Starting Grant)
Response
Dose-response testing with nanoparticles
Dose?
Tuning the test system…
NanoFM
FM
MW
LW
Radio Chemical
Plastic particles as model particles…
Lumbriculus variegatus (freshwater sediment worm). Aquatic
exposure to micro- and nanoplastics. Imaged by confocal microscopy
Control
1 µm PS-COOH
20 nm PS-COOH
Particle number pr volume or area
Nanoplastics – an environmental problem?
Mesh size cut-off
(e.g. 333 µm)
?
1 nm - 1µm
1µm – 1mm
Mesh size
1 mm - 1 cm
>1cm
Definition of nanoplastics
Proposed definitions
1 nm - 1µm
1µm – 1mm
67 -500 µm
Gregory & Andrady
2003
Browne et al
2007
Ryan et al
2009
Arthur et al
2009
Fendall and Sewell
2009
Costa et al
2010
EU Comission
2011
Hidalgo-Ruz et al
2012
Claessens et al
2013
<1mm
Van Cauwenberghe et al
2013
<1mm
Desforges et al
2014
>500 µm
1µm – 1mm
<1µm
<2mm
>1cm
1 mm - 1 cm
>5mm
>2cm
2 mm - 2 cm
<5mm
<5mm
>5mm
<1mm
1-100 nm
<5mm
1µm - 5mm
10-9
10-8
10-7
1 nm
nanoplastics
10-6
10-5
10-4
1 µm
microplastics
mesoplastics
macroplastics
10-3
10-2
1 mm
1 cm
particle size [m]
Some lessons learned from
nanoecotoxicology…
Interactions between nanoparticles and
algae (P. subcapitata) by transmission
electron microscopy (TEM)
A
B
C
Figure 3. Encapsulation of algal cells (Pseudokirchneriella subcapitata) exposed to TiO2
nanoparticles. A: Scanning electron microscopy (SEM) image of an algal cell after exposure to
50 mg/L TiO2 in ISO algal test media for 48 h. B: Corresponding SEM-EDX dot map shows
the distribution of Ti. It can be seen that the TiO2 nanoparticles cover the surface of the algae
(Modified from Hartmann et al., 2010 – Paper I). C: Transmission electron microscopy (TEM)
images showing the formation of algae-particle heteroaggregates (scale bar: 2 μm). (Modified
from Hartmann et al., 2011b – Paper IV)
Interactions between nanoplastics and algae
cells (P. subcapitata) by UV-VIS spectroscopy
and atomic force microscopy (AFM)
PS-COOH
PS-NH2
no. adsorbed particles
per algal cell
10000
1000
100
10
1
0
5
10
Ca2+ (mM)
15
Are effects caused by shading?
Indirect shading
Illumination
Control
Direct shading
0.9 ND filter
Violaxanthin
Lutein
Chlorophyll b
Chlorophyll a
Neoxanthin
Some lessons learned from
nanoecotoxicology…
Uptake and depuration of PS beads in
D.magna
Neonate D. magna (<24 h) exposed to 5 mg solids/L fluorescent
polystyrene beads (20 nm). Imaged by Fluorescent Light Sheet Microscopy
Control
1h
2h
4h
24h
Skjolding L. M., Ašmonaitė G., Jølck R. I., Baun A. and Sturve J., 2015, Uptake and
localization of fluorescent labelled nanoparticles in living crustaceans (Daphnia
magna) and zebrafish (Danio rerio) using Light Sheet Microscopy, manuscript
Uptake of PS beads in juvenile D. rerio –
aqueous and dietary exposure
3d
7d
Dietary exposure
(With 24h preexposed A. salina)
Aqueous exposure
1d
Skjolding L. M., Ašmonaitė G., Jølck R. I., Baun A. and Sturve J., 2015, Uptake and
localization of fluorescent labelled nanoparticles in living crustaceans (Daphnia
magna) and zebrafish (Danio rerio) using Light Sheet Microscopy, manuscript
Some lessons learned from
nanoecotoxicology…
Scenarios for particle interactions with copollutants
Unchanged bioavailability
of co-pollutant
Reduced
bioavailability
of co-pollutant
Reduced /increased
/unchanged depending
on particle and copollutant
Increased
bioavailability
of co-pollutant
‘Trojan horse’
‘Leaching’
Hartmann, NB, 2011, Ecotoxicity of engineered nanoparticles to freshwater organisms. PhD thesis, Technical
University of Denmark, Department of Environmental Engineering, ISBN: 978-87-92654-28-1
Some lessons learned from
nanoecotoxicology…
General test system considerations
• For meaningful dose-response testing we must:
– Control exposure
AND/OR
– Monitor/characterise exposure
Examples:
• Dispersion method matters!
• Media composition matters!
• Time matters!
Cupi, D., Hartmann, NB, Baun, A,
2015. Influence of pH and media
composition on stability and
nanoparticle ecotoxicity, manuscript
Sørensen, S.N. & Baun, A. (2014). Controlling silver nanoparticle exposure in
algal toxicity testing – a matter of timing. Nanotoxicology
10.3109/17435390.2014.913728
Exotoxicity testing of nanoplastics:
some key challenges
• Detection, identification and quantification in the environment
– Properties of environmentally weathered nanoplastics?
• Quantification of exposure in laboratory test
• Detection and quantification in test organisms
Options?
– Fluorescent dye
– Metal core (synthesis)
– C14 labelled?
– Chemical analysis of polymers
• Pyrolysis GC-MS?
• IR spectroscopy?
Thank you for your attention!
Acknowledgements:
Lars Michael Skjolding, DTU Environment
Signe Qualmann, DTU Environment
Basil Uthuppu, DTU Nanotech
Denisa Cupi, DTU Environment
Sara Nørgaard Sørensen, DTU Environment
European Research Council Starting Grant – ENV Nano
Contact:
Nanna B. Hartmann
DTU Environment, Technical University of Denmark
nibh@env.dtu.dk