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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.

Toxic and non-toxic polystyrene particles bind different proteins during filtration by zooplankton.

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.

Microplastics have recently become a growing environmental issue, whereas the smaller fractions, nanoplastics, have received less attention. Due to their small size, nanoplastics may affect organisms differently and potentially more severely than larger microplastics. In natural environments nanoplastics also interact with organic material and form larger aggregates, which may, potentially, reduce their toxicity as they grow in size. We tested the change in toxicity towards Daphnia magna by controlling the size of the aggregates of positively charged 50 nm polystyrene nanoplastics, which are highly toxic as single particles. We show that although 200 to 500 nm nanoplastics are not toxic, aggregates of 50 nm nanoplastics in the same size range are at least as toxic as the free, 50 nm, nanoplastics. Hence, an increase in size through aggregation, a process likely to occur as nanoparticles enter natural ecosystems, does not reduce toxicity. In a broader context this finding provides a firm basis for societal decision making regarding the potency of nanoparticles as they enter natural ecosystems.

Increasing human impact on the environment is causing drastic changes in disturbance regimes and how they prevail over time. Of increasing relevance is to further our understanding on biological responses to pulse disturbances (short duration) and how they interact with other ongoing press disturbances (constantly present). Because the temporal and spatial contexts of single experiments often limit our ability to generalize results across space and time, we conducted a modularized mesocosm experiment replicated in space (five lakes along a latitudinal gradient in Scandinavia) and time (two seasons, spring and summer) to generate general predictions on how the functioning and composition of multitrophic plankton communities (zoo-, phyto- and bacterioplankton) respond to pulse disturbances acting either in isolation or combined with press disturbances. As pulse disturbance, we used short-term changes in fish presence, and as press disturbance, we addressed the ongoing reduction in light availability caused by increased cloudiness and lake browning in many boreal and subarctic lakes. First, our results show that the top-down pulse disturbance had the strongest effects on both functioning and composition of the three trophic levels across sites and seasons, with signs for interactive impacts with the bottom-up press disturbance on phytoplankton communities. Second, community composition responses to disturbances were highly divergent between lakes and seasons: temporal accumulated community turnover of the same trophic level either increased (destabilization) or decreased (stabilization) in response to the disturbances compared to control conditions. Third, we found functional recovery from the pulse disturbances to be frequent at the end of most experiments. In a broader context, these results demonstrate that top-down, pulse disturbances, either alone or with additional constant stress upon primary producers caused by bottom-up disturbances, can induce profound but often functionally reversible changes across multiple trophic levels, which are strongly linked to spatial and temporal context dependencies. Furthermore, the identified dichotomy of disturbance effects on the turnover in community composition demonstrates the potential of disturbances to either stabilize or destabilize biodiversity patterns over time across a wide range of environmental conditions.

For aquatic scientists mesocosm experiments are important tools for hypothesis testing as they offer a compromise between experimental control and realism. Here we present a new mesocosm infrastructure-SITES AquaNET-located in five lakes connected to field stations in Sweden that cover a similar to 760 km latitudinal gradient. SITES AquaNet overcomes major hindrances in aquatic experimental research through: (i) openness to the scientific community, (ii) the potential to implement coordinated experiments across sites and time, and (iii) high-frequency measurements (temperature, photosynthetic photon flux density, turbidity and dissolved oxygen, chlorophyll a and phycocyanin concentrations) with an autonomous sensor system. Moreover, the infrastructure provides operational guidance and sensor expertise from technical staff, and connections to a multi-layered monitoring programme ("SITES Water") for each lake. This enables ecological observations from whole lake ecosystems to be compared with experimental studies aiming at disentangling major drivers and mechanisms underlying observed changes. Here we describe the technical properties of the infrastructure along with possibilities for experimental manipulations to tackle pressing issues in aquatic ecology and global change science. As a proof of concept, we also present a first mesocosm experiment across all five field sites with a cross-factorial design to evaluate responses of the sensor measurements to press/bottom-up (constant light reduction) and pulse/top-down (temporary fish predation) disturbances. This demonstrates the suitability of the infrastructure and autonomous sensor system to host modularized experiments and exemplifies the power and advantages of the approach to integrate a network of mecsocosm facilities with manageable costs across large geographic areas.

Ecological stability encompasses multiple dimensions of functional and compositional responses to environmental change. Though no single stability dimension used in isolation can fully reflect the overall response to environmental change, a common vulnerability assessment that integrates simultaneously across multiple stability components is highly desirable for ecological risk assessment. We develop both functional and compositional counterparts of a novel, integrative metric of overall ecological vulnerability (OEV). We test the framework with data from a modularized experiment replicated in five lakes over two seasons, examining functional and compositional responses to both pulse and press disturbances across three trophic groups. OEV is measured as the area under the curve integrated over the entire observation period, with the curve delimiting the difference between the disturbance treatment and undisturbed parallel controls, expressed either as the log response ratio of biomass (functional OEV) or community dissimilarity index (compositional OEV). Both, functional and compositional OEV correlated negatively with functional and compositional ‘resistance’, ‘temporal stability’ and ‘final/extent of recovery’ following both pulse and press disturbances, though less so with ‘resilience’ following a pulse disturbance. We also found a positive correlation between functional and compositional OEV, which reveals the potential to also evaluate the intricate linkage between biodiversity and functional change. Our findings demonstrate that OEV comprises a robust framework to: (a) capture simultaneously multiple functional and compositional stability components, and (b) quantify the functional consequences of biodiversity change. Our results provide the basis for an overarching framework for quantifying the overall vulnerability of ecosystems to environmental change, opening new possibilities for ecological risk assessment and management. Synthesis. Ecological stability comprises multiple dimensions that together encapsulate how ecosystems respond to environmental change. Considering these multiple aspects of stability simultaneously often poses a problem in environmental assessments, which frequently require overarching indicators of risk or vulnerability. While an analysis of multiple dimensions allows for deeper exploration of mechanisms, here we develop and test a new univariate indicator that integrates stability aspects under a broad range of disturbance regimes. Using a modularized experiment in Swedish lakes, we show that this integrative measure captures multiple stability dimensions reflecting compositional and functional vulnerability and their relationships between them.

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.

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.