Downloaded from orbit.dtu.dk on: Dec 31, 2016 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) Link to publication 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 General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal ? If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. 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 [email protected]
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