Abstract
Despite the increasing concern about the harmful effects of micro- and nanoplastics (MNPs), there are no harmonized guidelines or protocols yet available for MNP ecotoxicity testing. Current ecotoxicity studies often use commercial spherical particles as models for MNPs, but in nature, MNPs occur in variable shapes, sizes and chemical compositions. Moreover, protocols developed for chemicals that dissolve or form stable dispersions are currently used for assessing the ecotoxicity of MNPs. Plastic particles, however, do not dissolve and also show dynamic behavior in the exposure medium, depending on, for example, MNP physicochemical properties and the medium’s conditions such as pH and ionic strength. Here we describe an exposure protocol that considers the particle-specific properties of MNPs and their dynamic behavior in exposure systems. Procedure 1 describes the top-down production of more realistic MNPs as representative of MNPs in nature and particle characterization (e.g., using thermal extraction desorption-gas chromatography/mass spectrometry). Then, we describe exposure system development for short- and long-term toxicity tests for soil (Procedure 2) and aquatic (Procedure 3) organisms. Procedures 2 and 3 explain how to modify existing ecotoxicity guidelines for chemicals to target testing MNPs in selected exposure systems. We show some examples that were used to develop the protocol to test, for example, MNP toxicity in marine rotifers, freshwater mussels, daphnids and earthworms. The present protocol takes between 24 h and 2 months, depending on the test of interest and can be applied by students, academics, environmental risk assessors and industries.
Key points
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In nature, micro- and nanoplastics (MNPs) occur in various shapes, sizes and chemical compositions. Each of these properties can affect both their dynamic behavior and their toxicology and should be considered when performing ecotoxicology experiments to assess their risk.
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Here, MNPs are generated from plastic waste by using either a ball or centrifugal mill. Their physicochemical properties are measured. Ecotoxicology experiments in soil and aquatic systems are described.
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Acknowledgements
This work received financial support from the UEF Water research program, which is jointly funded by the Saastamoinen Foundation, the Wihuri Foundation and the Olvi Foundation. The study was also partially funded by the European Union’s Horizon 2020 research and innovation program, via the projects PLASTICSFATE (Grant Agreement number 965367) and POLYRISK (Grant Agreement number 964766). N.T. acknowledges the Canada Research Chair program and the Killam Research Fellowship.
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F.A.M. designed the protocol, and conceptualized and supervised the protocol development. All the co-authors contributed to developing, writing, editing and reviewing the protocol.
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Key references using this protocol
Jeong, C.-B. et al. Environ. Sci. Technol. 50, 8849–8857 (2016): https://doi.org/10.1021/acs.est.6b01441
Abdelsaleheen, O. et al. Sci. Total Environ. 798, 149196 (2021): https://doi.org/10.1016/j.scitotenv.2021.149196
Abdolahpur Monikh, F. et al. Nano Today 46, 101611 (2022): https://doi.org/10.1016/j.nantod.2022.101611
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Altmann, K. et al. Appl. Res. e202200078 (2023): https://doi.org/10.1002/appl.202200078
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Abdolahpur Monikh, F., Baun, A., Hartmann, N.B. et al. Exposure protocol for ecotoxicity testing of microplastics and nanoplastics. Nat Protoc 18, 3534–3564 (2023). https://doi.org/10.1038/s41596-023-00886-9
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DOI: https://doi.org/10.1038/s41596-023-00886-9
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Giovanni Di Guardo
Micro-nanoplastics as potential carriers of dioxins and Toxoplasma gondii in patients with carotid atheromas
Dear Editor,
The far more consistent risk of developing either a myocardial infarction or a stroke, if not even a fatal outcome, recently shown in patients affected by carotid atheromas carrying micro-nanoplastics (MNPs), compared with subjects harbouring no MNPs in the aforementioned lesions (1), is a major concern issue. Indeed, measurable levels of polyethylene and, to a lesser extent, also of polyvinyl chloride were detected in carotid atheromatous plaques from the former group of patients, with ultrastructural investigations additionally revealing the presence of foreign particles among plaque-infiltrating macrophages (1).
Notwithstanding the above, while additional knowledge could have been reasonably provided by the use of "ad hoc" animal models of cardiovascular disease (CVD) (2), the authors themselves admit there are several limitations in this work. Within such context, in fact, a number of endogenous and exogenous factors to which the individuals under study may have been exposed throughout their lives could justify, at least partially, the aforementioned differences in terms of CVD-associated/related morbidity and mortality. Furthermore, the possibility of a MNP contamination of carotid atheromas from the external environment, albeit remote, cannot be completely ruled out (1).
Apart from these objective limitations, it seems quite surprising, however, that the authors of this still absolutely relevant and interesting paper did not take into account the proven capability of MNPs to act as powerful "attractors and concentrators" for a huge number of persistent environmental pollutants such as dioxins and for several microbial pathogens including Toxoplasma gondii (3), a zoonotic protozoan agent for which a close interaction with polyethylene microbeads and polyester microfibers has been recently described (4).
As a matter of fact, while dioxin exposure has been linked to CVD mortality (5), T. gondii infection may be also associated with myocarditis, constrictive pericarditis, pericardial effusion, arrhythmias (both atrial and ventricular) and congestive/acute heart failure (6).
Therefore, additional work paying special attention to the highly complex and multifaceted "chemico-microbiological profile" of MNPs should be strongly encouraged in the next future, with the results originating from this absolutely necessary research effort hopefully allowing to assess "who does what" in exacerbating a preexisting CVD condition.
And, while cumulative evidence has also shown that the inappropriate disposal of face masks and plastic gloves used in the fight against the pandemic SARS-CoV-2 betacoronavirus has resulted in a growing MNP contamination of terrestrial, sea and ocean ecosystems on a global scale (7),
an integrated multidisciplinary, holistic and One Health-based approach would be highly recommendable when dealing with the still largely unknown MNP effects on human, animal and environmental health.
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Giovanni Di Guardo, DVM, Dipl. ECVP,
Former Professor of General Pathology and Veterinary Pathophysiology at the Veterinary Medical Faculty of the University of Teramo,
Località Piano d'Accio, 64100 Teramo, Italy
Private address: Viale Pasteur, 77
00144 - EUR - Rome, Italy
(E-mail address: gdiguardo@unite.it;
cellular phone no.: +39-347-6317862)