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Aquatic ecotoxicity testing of nanoplasticslessons learned from nanoecotoxicology

Hartmann, Nanna Isabella Bloch; Nolte, Tom; A. Sørensen, Mads; R. Jensen, Pernille; Baun, Anders

Publication date:2015

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Citation (APA):Hartmann, N. I. B. (Author), Nolte, T. (Author), A. Sørensen, M. (Author), R. Jensen, P. (Author), & Baun, A.(Author). (2015). Aquatic ecotoxicity testing of nanoplastics: lessons learned from nanoecotoxicology.Sound/Visual production (digital), DTU Environment.

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

Hazard Identification

What do we do?

MARINA

EnvNano

NanoDen

New paradigm for ecotoxicity testing of NMs (European Research Council Starting Grant)

NanoREM

Dose-response testing with nanoparticles

Dose?

Res

pons

e

NanoFM Radio Chemical FM MW LW

Tuning the test system…

Plastic particles as model particles…

Control 20 nm PS-COOH 1 µm PS-COOH

Lumbriculus variegatus (freshwater sediment worm). Aquatic exposure to micro- and nanoplastics. Imaged by confocal microscopy

Nanoplastics – an environmental problem?

1 nm - 1µm 1µm – 1mm 1 mm - 1 cm >1cm

?

Mesh size

Part

icle

num

ber

pr v

olum

e or

are

a Mesh size cut-off (e.g. 333 µm)

Definition of nanoplastics

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

Van Cauwenberghe et al 2013

Desforges et al 2014

10-9 10-8 10-7 10-6 10-5 10-4 10-3 10-2 particle size [m]

1 nm 1 µm 1 mm 1 cm

>5mm 1µm – 1mm

<2mm >2cm 2 mm - 2 cm

<5mm <5mm

<1mm

1-100 nm

>5mm

<5mm

<1mm

<1mm

1µm - 5mm

microplastics nanoplastics mesoplastics macroplastics

Proposed definitions 1 nm - 1µm 1µm – 1mm 1 mm - 1 cm >1cm

67 -500 µm >500 µm

<1µm

Some lessons learned from nanoecotoxicology…

Interactions between nanoparticles and algae (P. subcapitata) by transmission electron microscopy (TEM)

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)

C

A B

Interactions between nanoplastics and algae cells (P. subcapitata) by UV-VIS spectroscopy and atomic force microscopy (AFM)

1

10

100

1000

10000

0 5 10 15

no.

ad

sorb

ed p

arti

cles

p

er a

lgal

cel

l

Ca2+ (mM)

PS-COOH PS-NH2

Are effects caused by shading?

ViolaxanthinLuteinChlorophyll bChlorophyll aNeoxanthin

Control 0.9 ND filter

Direct shading Illumination

Indirect shading

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

1h 2h 4h 24h Control

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

1d 3d 7d

Aqu

eous

exp

osur

e D

ieta

ry e

xpos

ure

(With

24h

pre

-ex

pose

d A.

salin

a)

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 co-pollutants

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

Reduced bioavailability of co-pollutant

Unchanged bioavailability of co-pollutant

Increased bioavailability of co-pollutant

‘Trojan horse’

Reduced /increased /unchanged depending on particle and co-pollutant ‘Leaching’

Some lessons learned from nanoecotoxicology…

• For meaningful dose-response testing we must: – Control exposure AND/OR – Monitor/characterise exposure

Examples: • Dispersion method matters! • Media composition matters! • Time matters!

General test system considerations

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

• 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?

Exotoxicity testing of nanoplastics: some key challenges

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