Altered behavior, physiology, and metabolism in fish exposed to polystyrene nanoparticles.

The use of nanoparticles in consumer products, for example, cosmetics, sunscreens, and electrical devices, has increased tremendously over the past decade despite insufficient knowledge about their effects on human health and ecosystem function. Moreover, the amount of plastic waste products that enter natural ecosystems, such as oceans and lakes, is increasing, and degradation of the disposed plastics produces smaller particles toward the nano scale. Therefore, it is of utmost importance to gain knowledge about how plastic nanoparticles enter and affect living organisms. Here we have administered 24 and 27 nm polystyrene nanoparticles to fish through an aquatic food chain, from algae through Daphnia, and studied the effects on behavior and metabolism. We found severe effects on feeding and shoaling behavior as well as metabolism of the fish; hence, we conclude that polystyrene nanoparticles have severe effects on both behavior and metabolism in fish and that commonly used nanosized particles may have considerable effects on natural systems and ecosystem services derived from them.

Induced tolerance expressed as relaxed behavioural threat response in millimetre-sized aquatic organisms.

Natural selection shapes behaviour in all organisms, but this is difficult to study in small, millimetre-sized, organisms. With novel labelling and tracking techniques, based on nanotechnology, we here show how behaviour in zooplankton (Daphnia magna) is affected by size, morphology and previous exposure to detrimental ultraviolet radiation (UVR). All individuals responded with immediate downward swimming to UVR exposure, but when released from the threat they rapidly returned to the surface. Large individuals swam faster and generally travelled longer distances than small individuals. Interestingly, individuals previously exposed to UVR (during several generations) showed a more relaxed response to UVR and travelled shorter total distances than those that were naive to UVR, suggesting induced tolerance to the threat. In addition, animals previously exposed to UVR also had smaller eyes than the naive ones, whereas UVR-protective melanin pigmentation of the animals was similar between populations. Finally, we show that smaller individuals have lower capacity to avoid UVR which could explain patterns in natural systems of lower migration amplitudes in small individuals. The ability to change behavioural patterns in response to a threat, in this case UVR, adds to our understanding of how organisms navigate in the 'landscape of fear', and this has important implications for individual fitness and for interaction strengths in biotic interactions.

Environmental risks of breakdown nanoplastics from synthetic football fields.

The widespread use of synthetic turf in sports has raised health concerns due to potential risks from nanoplastic inhalation or ingestion. Our research focused on detecting nanoplastics in drainage water from a synthetic football field and evaluating the toxicity of these materials after mechanical fragmentation. We collected and analysed drainage water samples for polymer content and subjected high-density polyethylene (HDPE) straws and ethylene propylene diene monomer (EPDM) granules used on synthetic football fields, to mechanical breakdown to create nanoplastics. The results indicated the presence of trace amounts of EPDM in the water samples. Furthermore, the toxicological assessment revealed that the broken-down nanoplastics and leachate from the surface of EPDM rubber granules exhibited high toxicity to Daphnia magna, while nanoplastics from the inner material exhibited no significant toxicity. The findings highlight the urgent need for future research to identify these specific toxic agents from the surface of EPDM granules.

Prolonged survival time of Daphnia magna exposed to polylactic acid breakdown nanoplastics.

Polylactic acid nanoparticles (PLA NPs) according to food and drug administration are biodegradable and biocompatible polymers that have received a lot of attention due to their natural degradation mechanism. Although there is already available information concerning the effects of PLA microplastic to aquatic organisms, the knowledge about PLA NPs is still vague. In the present study, we analyzed the chemical composition of engineered PLA NPs, daily used PLA items and their breakdown products. We show that PLA breakdown products are oxidized and may contain aldehydes and/or ketones. The breakdown produces nanosized particles, nanoplastics, and possibly other small molecules as lactide or cyclic oligomers. Further, we show that all PLA breakdown nanoplastics extended the survival rate in Daphnia magna in an acute toxicity assay, however, only PLA plastic cup breakdown nanoplastics showed a significant difference compared to a control group.

Nanoplastics released from daily used silicone and latex products during mechanical breakdown.

Waste of polymer products, especially plastics, in nature has become a problem that caught the awareness of the general public during the last decade. The macro- and micro polymers in nature will be broken down by naturally occurring events such as mechanical wear and ultra-violet (UV) radiation which will result in the generation of polymeric particles in the nano-size range. We have recently shown that polystyrene and high-density polyethylene macroplastic can be broken down into nano-sized particles by applying mechanical force from an immersion blender. In this article, we show that particles in the nano-size range are released from silicone and latex pacifiers after the same treatment. Additionally, boiling the pacifiers prior to the mechanical breakdown process results in an increased number of particles released from the silicone but not the latex pacifier. Particles from the latex pacifier are acutely toxic to the freshwater filter feeding zooplankter Daphnia magna.

Characterization and Toxic Potency of Airborne Particles Formed upon Waste from Electrical and Electronic Equipment Waste Recycling: A Case Study.

Manual dismantling, shredding, and mechanical grinding of waste from electrical and electronic equipment (WEEE) at recycling facilities inevitably lead to the accidental formation and release of both coarse and fine particle aerosols, primarily into the ambient air. Since diffuse emissions to air of such WEEE particles are not regulated, their dispersion from the recycling plants into the adjacent environment is possible. The aim of this interdisciplinary project was to collect and characterize airborne WEEE particles smaller than 1 µm generated at a Nordic open waste recycling facility from a particle concentration, shape, and bulk and surface composition perspective. Since dispersed airborne particles eventually may reach rivers, lakes, and possibly oceans, the aim was also to assess whether such particles may pose any adverse effects on aquatic organisms. The results show that WEEE particles only exerted a weak tendency toward cytotoxic effects on fish gill cell lines, although the exposure resulted in ROS formation that may induce adverse effects. On the contrary, the WEEE particles were toxic toward the crustacean zooplankter Daphnia magna, showing strong effects on survival of the animals in a concentration-dependent way.

Size fractionation of high-density polyethylene breakdown nanoplastics reveals different toxic response in Daphnia magna.

Plastic litter is a growing environmental problem. Recently, microplastics and nanoplastics, produced during breakdown processes in nature, have been in focus. Although there is a growing knowledge concerning microplastic, little is still known about the effect of nanoplastics. We have showed that mechanical breakdown of high-density polyethylene (HDPE), followed by filtration through 0.8 µm filters, produces material toxic to the freshwater zooplankton Daphnia magna and affected the reproduction in life-time tests. However, further size fractionation and purification reveals that the nanoplastics fraction is non-toxic at these concentrations, whereas the fraction with smaller sizes, below ~ 3 nm, is toxic. The HDPE nanoplastics are highly oxidized and with an average diameter of 110 nm. We conclude that mechanical breakdown of HDPE may cause environmental problems, but that the fraction of leached additives and short chain HDPE are more problematic than HDPE nanoplastics.

Review of ecotoxicological studies of widely used polystyrene nanoparticles.

With polystyrene nanoparticles being widely used in various applications, there is a great need for deeper knowledge on the safety, fate and biological effects of these particles on both individual living organisms and the whole ecosystems. Due to this, there is a growing interest in performing ecotoxicological studies using model plastic nanoparticles, and consequently it generates an increasing number of published papers describing the negative impact on wildlife caused by such nanoparticles. Polystyrene is the most studied nanosized plastic, therefore this review focuses on research conducted with manufactured polystyrene nanoparticles. The aim of the present article is to provide a critical methodological outline of the existing ecotoxicological studies on the effects of polystyrene nanoparticles on aquatic organisms. Going through the published articles, we noted that particle characterization especially in the test medium, can be improved. The analysis also highlights the importance of purifying the polystyrene nanoparticles before studying its toxicity. Furthermore, the size characterization of such nanoparticles is underemphasized, and in future studies, authors should consider including more techniques to achieve this goal. Finally, short-term or direct exposure scenarios do not add the most environmentally relevant knowledge in terms of the toxicity caused by polystyrene nanoparticles.

Adsorption of bio-organic eco-corona molecules reduces the toxic response to metallic nanoparticles in Daphnia magna.

As the use of engineered nanomaterials increases, so does the risk of them spreading to natural ecosystems. Hitherto, knowledge regarding the toxic properties of nanoparticles (NP's) and their potential interactions with natural bio-organic molecules adsorbed to them, and thereby forming surface coronas, is limited. However, we show here that the toxic effect of NPs of tungsten carbide cobalt (WC-Co) and cobalt (Co) on the crustacean Daphnia magna is postponed in the presence of natural biological degradation products (eco-corona biomolecules). For Daphnia exposed to WC-Co NPs the survival time increased with 20-25% and for Co NPs with 30-47% after mixing the particles with a solution of eco-corona biomolecules before exposure. This suggests that an eco-corona, composed of biomolecules always present in natural ecosystems, reduces the toxic potency of both studied NPs. Further, the eco-coronas did not affect the particle uptake, suggesting that the reduction in toxicity was related to the particle-organism interaction after eco-corona formation. In a broader context, this implies that although the increasing use and production of NPs may constitute a novel, global environmental threat, the acute toxicity and long-term effects of some NPs will, at least under certain conditions, be reduced as they enter natural ecosystems.

Long-term exposure to nanoplastics reduces life-time in Daphnia magna.

Plastics are widely used in todays society leading to an accelerating amount of plastic waste entering natural ecosystems. Over time these waste products degrade to micro- and, eventually, nanoplastic particles. Therefore, the break-down of plastics may become a critical threat to aquatic ecosystems and several short term studies have demonstrated acute toxicity of nanoplastics on aquatic organisms. However, our knowledge about effects of chronic or life-time exposure on freshwater invertebrates remains elusive. Here, we demonstrate results from life-time exposure (103 days) of a common freshwater invertebrate, Daphnia magna, exposed to sub-lethal concentrations of polystyrene nanoparticles. 53 nm positively charged aminated polystyrene particles were lethal at concentration of 0.32 mg/L which is two magnitudes lower than previously used concentrations in short-term (24 h) tests. At this concentration the life-time of individuals was shortened almost three times. Negatively charged carboxylated 26 and 62 nm polystyrene particles, previously demonstrated to be non-toxic at 25 and 50 mg/L concentrations in short-term tests, were toxic to D. magna at all concentrations used in our long-term study. Although total reproductive output was not significantly affected at increasing concentrations of polystyrene nanoparticles, there was a decreasing trend in the number of offspring over their life-time. Hence, in order to understand how the potential future environmental problem of nanoplastic particles may affect biota, long-term or life-time studies resembling environmental concentrations should be performed in order to provide information for predictions of future scenarios in natural aquatic environments.

Nanoplastics formed during the mechanical breakdown of daily-use polystyrene products.

Large amounts of plastics are released into the environment every day. These released plastics have a clearly documented negative effect on wildlife. Much research attention has been given to large plastic pieces and microplastics. However, if the breakdown of plastics is a continous process, eventually nanoplastics will be produced. Nanoplastics will affect wildlife differently from larger plastic pieces. We have studied the products formed by the mechanical breakdown of two commonly used polystyrene products, takeaway coffee cup lids and expanded polystyrene foam. After breakdown using a food processor, we characterized the breakdown products using seven different methods and found nanosized polystyrene particles with different shapes and negative or nearly neutral surface charges. These results clearly demonstrate that daily-use polystyrene products can break down into nanoparticles. Model polystyrene particles with different sizes and surface modifications have previously been shown to have different negative effects on wildlife. This indicates that breakdown nanoparticles might have the potential to cause cocktail effects in nature.

Long-term effects of tungsten carbide (WC) nanoparticles in pelagic and benthic aquatic ecosystems.

As the production and usage of nanomaterials are increasing so are the concerns related to the release of the material into nature. Tungsten carbide (WC) is widely used for its hard metal properties, although its use, in for instance tyre studs, may result in nano-sized particles ending up in nature. Here, we evaluate the potential long-term exposure effects of WC nanoparticles on a pelagic (Daphnia magna) and a benthic (Asellus aquaticus) organism. No long-term effects were observed in the benthic system with respect to population dynamics or ecosystem services. However, long-term exposure of D. magna resulted in increased time to first reproduction and, if the particles were resuspended, strong effects on survival and reproductive output. Hence, the considerable differences in acute vs. long-term exposure studies revealed here emphasize the need for more long-term studies if we are to understand the effects of nanoparticles in natural systems.

Translocation of 40 nm diameter nanowires through the intestinal epithelium of Daphnia magna.

Nanowires (NWs) have unique electrical and optical properties of value for many applications including lighting, sensing, and energy harnessing. Consumer products containing NWs increase the risk of NWs being released in the environment, especially into aquatic ecosystems through sewage systems. Daphnia magna is a common, cosmopolitan freshwater organism sensitive to toxicity tests and represents a likely entry point for nanoparticles into food webs of aquatic ecosystems. Here we have evaluated the effect of NW diameter on the gut penetrance of NWs in Daphnia magna. The animals were exposed to NWs of two diameters (40 and 80 nm) and similar length (3.6 and 3.8 µm, respectively) suspended in water. In order to locate the NWs in Daphnia, the NWs were designed to comprise one inherently fluorescent segment of gallium indium phosphide (GaInP) flanked by a gallium phosphide (GaP) segment. Daphnia mortality was assessed directly after 24 h of exposure and 7 days after exposure. Translocation of NWs across the intestinal epithelium was investigated using confocal fluorescence microscopy directly after 24 h of exposure and was observed in 89% of Daphnia exposed to 40 nm NWs and in 11% of Daphnia exposed to 80 nm NWs. A high degree of fragmentation was observed for NWs of both diameters after ingestion by the Daphnia, although 40 nm NWs were fragmented to a greater extent, which could possibly facilitate translocation across the intestinal epithelium. Our results show that the feeding behavior of animals may enhance the ability of NWs to penetrate biological barriers and that penetrance is governed by the NW diameter.

Three-dimensional tracking of small aquatic organisms using fluorescent nanoparticles.

Tracking techniques are vital for the understanding of the biology and ecology of organisms. While such techniques have provided important information on the movement and migration of large animals, such as mammals and birds, scientific advances in understanding the individual behaviour and interactions of small (mm-scale) organisms have been hampered by constraints, such as the sizes of existing tracking devices, in existing tracking methods. By combining biology, chemistry and physics we here present a method that allows three-dimensional (3D) tracking of individual mm-sized aquatic organisms. The method is based on in-vivo labelling of the organisms with fluorescent nanoparticles, so-called quantum dots, and tracking of the organisms in 3D via the quantum-dot fluorescence using a synchronized multiple camera system. It allows for the efficient and simultaneous study of the behaviour of one as well as multiple individuals in large volumes of observation, thus enabling the study of behavioural interactions at the community scale. The method is non-perturbing - we demonstrate that the labelling is not affecting the behavioural response of the organisms - and is applicable over a wide range of taxa, including cladocerans as well as insects, suggesting that our methodological concept opens up for new research fields on individual behaviour of small animals. Hence, this offers opportunities to focus on important biological, ecological and behavioural questions never before possible to address.