Cumulative Impacts of Oil Pollution, Ocean Warming, and Coastal Freshening on the Feeding of Arctic Copepods.

The Arctic is undergoing rapid changes, and biota are exposed to multiple stressors, including pollution and climate change. Still, little is known about their joint impact. Here, we investigated the cumulative impact of crude oil, warming, and freshening on the copepod species Calanus glacialis and Calanus finmarchicus. Adult females were exposed to ambient conditions (control; 0 °C + 33 psu) and combined warming and freshening: 5 °C + 27 psu (Scenario 1), 5 °C + 20 psu (Scenario 2) for 6 days. All three conditions were tested with and without dispersed crude oil. In Scenario 1, fecal pellet production (FPP) significantly increased by 40-78% and 42-122% for C. glacialis and C. finmarchicus, respectively. In Scenario 2, FPP decreased by 6-57% for C. glacialis, while it fluctuated for C. finmarchicus. For both species, oil had the strongest effect on FPP, leading to a 68-83% reduction. This overshadowed the differences between climatic scenarios. All variables (temperature, salinity, and oil) had significant single effects and several joint effects on FPP. Our results demonstrate that Arctic copepods are sensitive to environmentally realistic concentrations of crude oil and climate change. Strong reductions in feeding can reduce the copepods' energy content with potential large-scale impacts on the Arctic marine food web.

The Behavior of Planktonic Copepods Minimizes the Entry of Microplastics in Marine Food Webs.

The entry of microplastics (MPs) into marine food webs is a major environmental concern. We investigated how the behavior of planktonic copepods influences the risk of MPs to enter marine food webs by applying a trait-based approach and by combining experiments (bottle incubations and video observations) with biogeographical analyses. We aimed to evaluate which type of feeding behavior is most risky in terms of MP ingestion and which marine geographical areas are more susceptible to MP ingestion by planktonic copepods. We used different species as models of the main foraging behaviors in planktonic copepods: feeding-current, cruising, ambush, and mixed behavior feeding. All behaviors showed a similarly low risk of MP ingestion, up to 1 order of magnitude lower than for similar-sized microalgae. We did not observe any influence of the prey type or MP size (8 and 20 µm) on MP ingestion for any of the behaviors. By mapping the global distribution of feeding behaviors, we showed that feeding-current feeding is the most common behavior, but the risk of MP ingestion remains equally low across the global ocean, independently of the predominant behavior. Overall, our results suggest a low risk of MP ingestion by planktonic copepods and therefore a minimal risk of trophic transfer of MPs via marine pelagic copepods in marine ecosystems.

Crude Oil and Its Burnt Residues Induce Metamorphosis in Marine Invertebrates.

Metamorphosis is a critical process in the life cycle of most marine benthic invertebrates, determining their transition from plankton to benthos. It affects dispersal and settlement and therefore decisively influences the dynamics of marine invertebrate populations. An extended period of metamorphic competence is an adaptive feature of numerous invertebrate species that increases the likelihood of finding a habitat suitable for settlement and survival. We found that crude oil and residues of burnt oil rapidly induce metamorphosis in two different marine invertebrate larvae, a previously unknown sublethal effect of oil pollution. When exposed to environmentally realistic oil concentrations, up to 84% of tested echinoderm larvae responded by undergoing metamorphosis. Similarly, up to 87% of gastropod larvae metamorphosed in response to burnt oil residues. This study demonstrates that crude oil and its burned residues can act as metamorphic inducers in marine planktonic larvae, short-circuiting adaptive metamorphic delay. Future studies on molecular pathways and oil-bacteria-metamorphosis interactions are needed to fully understand the direct or indirect mechanisms of oil-induced metamorphosis in marine invertebrates. With 90% of chronic oiling occurring in coastal areas, this previously undescribed impact of crude oil on planktonic larvae may have global implications for marine invertebrate populations and biodiversity.

Shapes of Hyperspectral Imaged Microplastics.

Shape matters for microplastics, but its definition, particularly for hyperspectral imaged microplastics, remains ambiguous and inexplicit, leading to incomparability across data. Hyperspectral imaging is a common approach for quantification, yet no unambiguous microplastic shape classification exists. We conducted an expert-based survey and proposed a set of clear and concise shapes (fiber, rod, ellipse, oval, sphere, quadrilateral, triangle, free-form, and unidentifiable). The categories were validated on images of 11,042 microplastics from four environmental compartments (seven matrices: indoor air; wastewater influent, effluent, and sludge; marine water; stormwater; and stormwater pond sediments), by inviting five experts to score each shape. We found that the proposed shapes were well defined, representative, and distinguishable to the human eye, especially for fiber and sphere. Ellipse, oval, and rod were though less distinguishable but dominated in all water and solid matrices. Indoor air held more unidentifiable, an abstract shape that appeared mostly for particles below 30 µm. This study highlights the need for assessing the recognizability of chosen shape categories prior to reporting data. Shapes with a clear and stringent definition would increase comparability and reproducibility across data and promote harmonization in microplastic research.

Unpalatable Plastic: Efficient Taste Discrimination of Microplastics in Planktonic Copepods.

Planktonic copepods are the most abundant animals in the ocean and key players in global biochemical processes. Recent modeling suggests that zooplankton ingestion of microplastics (MPs) can disrupt the biological carbon pump and accelerate a global loss of oceanic oxygen. Here we investigate the behavioral responses and ingestion rates of a model feeding-current generating copepod when exposed to microplastics of different characteristics by small-scale video observations and bottle incubations. We found that copepods rejected 80% of the microplastics after touching them with their mouth parts, in essence exhibiting a kind of taste discrimination. High rejection rates of microplastics were independent of polymer type, shape, presence of biofilms, or sorbed pollutant (pyrene), indicating that microplastics are unpalatable for feeding-current feeding copepods and that post-capture taste discrimination is a main sensorial mechanism in the rejection of microplastics. In an ecological context, taking into account the behaviors of planktonic copepods and the concentrations of microplastics found in marine waters, our results suggest a low risk of microplastic ingestion by zooplankton and a low impact of microplastics on the vertical exportation of fecal pellets.

Are We Speaking the Same Language? Recommendations for a Definition and Categorization Framework for Plastic Debris.

The accumulation of plastic litter in natural environments is a global issue. Concerns over potential negative impacts on the economy, wildlife, and human health provide strong incentives for improving the sustainable use of plastics. Despite the many voices raised on the issue, we lack a consensus on how to define and categorize plastic debris. This is evident for microplastics, where inconsistent size classes are used and where the materials to be included are under debate. While this is inherent in an emerging research field, an ambiguous terminology results in confusion and miscommunication that may compromise progress in research and mitigation measures. Therefore, we need to be explicit on what exactly we consider plastic debris. Thus, we critically discuss the advantages and disadvantages of a unified terminology, propose a definition and categorization framework, and highlight areas of uncertainty. Going beyond size classes, our framework includes physicochemical properties (polymer composition, solid state, solubility) as defining criteria and size, shape, color, and origin as classifiers for categorization. Acknowledging the rapid evolution of our knowledge on plastic pollution, our framework will promote consensus building within the scientific and regulatory community based on a solid scientific foundation.

Toxicity of tire particle leachates on early life stages of keystone sea urchin species.

Particles from tires are a major fraction of microplastic pollution. They contain a wide range of chemical additives that can leach into the water and be harmful to aquatic organisms. In this study, we investigated the acute toxicity of tire particle leachates in early life stages of three keystone echinoderm species (Paracentrotus lividus, Arbacia lixula, Diadema africanum). Embryos were exposed for 72 h to a range of leachate dilutions, prepared using a concentration of 1 g L-1. Larval growth, abnormal development, and mortality were the measured endpoints. Furthermore, we estimated the activity of glutathione S transferase (GST) and the electron transport system (ETS) in P. lividus. Strong concentration-dependent responses were observed in all species, though with differing sensitivity. The median effect concentrations for abnormal development in P. lividus and A. lixula were 0.16 and 0.35 g L-1, respectively. In D. africanum, mortality overshadowed abnormal development and the median lethal concentration was 0.46 g L-1. Larvae of P. lividus were significantly smaller than the control from 0.125 g L-1, while the other two species were affected from 0.5 g L-1. ETS activity did not change but there was a non-significant trend of increasing GST activity with leachate concentration in P. lividus. Seven organic chemicals and eight metals were detected at elevated concentrations in the leachates. While we regard zinc as a strong candidate to explain some of the observed toxicity, it can be expected that tire particle leachates exhibit a cocktail effect and other leached additives may also contribute to their toxicity. Our results emphasize the importance of multi-species studies as they differ in their susceptibility to tire particle pollution. We found negative effects at concentrations close to projections in the environment, which calls for more research and mitigation actions on these pollutants.

How fast, how far: Diversification and adoption of novel methods in aquatic microplastic monitoring.

Since 2004, there has been a marked diversification in the methods used to determine aquatic microplastic (MP) concentrations. Despite calls for a unified approach to MP sampling, the proliferation of new methods has accelerated in recent years. Both minor method adaptations and entirely novel approaches have been introduced to overcome barriers to reliable MP sampling, extraction and quantification, resulting in a variety of complimentary but also competing approaches. However, there is little clarity regarding the extent to which new methods are acknowledged and adopted, or of the apparent drivers of, as well as barriers to, said adoption. To explore these issues, the rate of method diversification was examined in a systematic review. The rate and degree of diversification were determined by scoring each method by its "degree of novelty": highly novel methods, secondary adaptations of existing methods and smaller, tertiary adaptations of existing methods. This analysis revealed that the rate of method diversification has been greatest since 2011. Our results indicate limited use of these novel methods and adaptations in the subsequent literature, with many researchers falling back on methods that are well established in the existing literature. Importantly, there is little consistency in the units used when reporting MP concentrations. However, these differences are seldom driven by method selection and are rather the result of discrepancies between researchers. Thus, in understanding the requirements of comparability and consistent reporting for monitoring purposes, we can apply a diverse approach to sampling whilst maintaining the applicability and usefulness of the resulting data.

Quantification of plankton-sized microplastics in a productive coastal Arctic marine ecosystem.

Microplastics (MPs) are polluting the Arctic, but our understanding of their abundance, distribution, and sources is limited. This study quantified MPs down to 10 µm in marine waters of the most populated region in Greenland. A new plastic-free pump-filter system was used to collect MPs from surface waters in the fjord Nuup Kangerlua close to Nuuk. Additionally, we took samples by horizontal tows with a bongo net (300 µm mesh-size). The median concentrations were 142 MPs m-3 and 0.12 MPs m-3 in the pump and bongo samples, respectively. The most abundant polymer was polyester across stations and sampling types. Fibers were the dominant shape in the bongo samples, while non-fibrous particles dominated in the pump samples. MP abundance was lower in the fjord and increased close to Nuuk and towards the open ocean, indicating that Nuuk is an important point source for MPs. In both samples, concentrations of MPs increased with decreasing size, illustrating the importance of quantifying the smallest fraction of MPs. Thus, the use of methods allowing for a quantification of the smallest MPs is vital to reduce the underestimation of MP concentrations in the environment. The smallest size fraction is also most available to plankton-feeding marine invertebrates and an important entry point for MPs into marine food webs. At the found concentrations, immediate adverse effects on the pelagic food webs are unlikely. However, growing anthropogenic activities could increase the risk of MPs to affect the sensitive Arctic ecosystem.

Ingestion and impact of microplastics on arctic Calanus copepods.

Microplastics (MPs) are contaminants of emerging concern in the Arctic, but knowledge of their potential effects on Arctic plankton food webs remains scarce. We experimentally investigated ingestion and effects of MPs (20 µm polyethylene spheres) on the arctic copepods Calanus finmarchicus, C. glacialis and C. hyperboreus. These species dominate arctic zooplankton biomass and are relevant target species to investigate the potential impacts of MPs on the Arctic marine ecosystem. Females of each species were exposed to two concentrations of MPs (200 and 20,000 MPs L-1) in combination with different food (diatom) concentrations, reflecting high (3000-5000 cells mL-1, spring phytoplankton bloom) and low (50-500 cells mL-1, pre/post bloom) food conditions. MPs did not affect negatively fecal pellet production rates in any of the species at the studied exposure concentrations. However, egg production rates of copepods exposed to MPs were 8 times higher compared with the controls, which suggests that MP exposure can cause stress-induced spawning in arctic copepods. Microscopic examination of the fecal pellets confirmed ingested MPs in the three species (up to aprox. 1000 MPs cop-1 d-1). The number of MPs per pellet decreased exponentially with increasing food concentration. The daily ingestion of MPs per copepod was higher at low- food concentrations (250-500 cells mL-1). At our exposure conditions, the presence of MPs inside C. hyperboreus fecal pellets did not affect their sinking rates. Overall, our experimental research show that 1) acute exposure to virgin polyethylene MPs has a low impact on arctic Calanus species at environmentally relevant MP concentrations, independent of food availability, and 2) arctic copepods influence the environmental fate of plankton-sized MPs by exporting buoyant MPs from the surface layer to the sea floor via fecal pellets.

Ingestion and effects of micro- and nanoplastics in blue mussel (Mytilus edulis) larvae.

It is well known that mussels are exposed to microplastics but ingestion and potential effects on mussel larvae are not well understood. We quantified ingestion and egestion of 100 nm and 2 µm polystyrene beads in blue mussel larvae after 4 h exposure and 16 h depuration using different plastic-to-microalgae ratios. Effects on growth and development of mussel larvae were investigated at 0.42, 28.2 and 282 µgL-1 within 15 days of exposure. We found that, on a mass basis, larvae ingested a higher amount of 2 µm than 100 nm beads, while egestion was independent of particle size and the plastics-to-algae ratio. Although particle egestion occurred readily, microplastics remained inside the larvae. Larval growth was not affected but abnormally developed larvae increased after exposure to polystyrene beads. Malformations were more pronounced for 100 nm beads, at higher concentration and after longer exposure time.

When Fluorescence Is not a Particle: The Tissue Translocation of Microplastics in Daphnia magna Seems an Artifact.

Previous research reported the translocation of nano- and microplastics from the gastrointestinal tract to tissues in Daphnia magna, most prominently of fluorescent polystyrene beads to lipid droplets. For particles >300 nm, such transfer is biologically implausible as the peritrophic membrane retains these in the daphnid gut. We used confocal laser scanning microscopy to study tissue transfer applying the setup from a previous study (neonates exposed to 20 and 1000 nm polystyrene beads at 2 µg L-1 for 4 and 24 h), the same setup with a fructose-based clearing, and a setup with a 1000-fold higher concentration (2 mg L-1 ). We used passive sampling to investigate whether the beads leach the fluorescent dye. Although the 1000 nm beads were visible in the gut at both exposure concentrations, the 20 nm beads were detectable at 2 mg L-1 only. At this concentration, we observed fluorescence in lipid droplets in daphnids exposed to both particle types. However, this did not colocalize with the 1000 nm beads, which remained visible in the gut. We further confirmed the leaching of the fluorescent dye using a passive sampler, a method that can also be applied in future studies. In summary, we cannot replicate the original study but demonstrate that the fluorescence in the lipid droplets of D. magna results from leaching of the dye. Thus, the use of fluorescence as a surrogate for particles can lead to artifacts in uptake and translocation studies. This highlights the need to confirm the stability of the fluorescence label or to localize particles using alternative methods. Environ Toxicol Chem 2019;38:1495-1503. © 2019 SETAC OPEN PRACTICES: The present study has earned Open Data/Materials badges for making publicly available the digitally shareable data necessary to reproduce the reported results. Learn more about the Open Practices badges from the Center for Open Science: https://osf.io/tvyxz/wiki.

The fate of microplastics during uptake and depuration phases in a blue mussel exposure system.

We present a blue mussel exposure system where the fate of microplastics (polystyrene beads) is tracked during exposure and depuration phases. This enabled the establishment of a complete mass balance. Quantification of beads in mussels was done with a novel enzymatic digestion protocol. We found a similar relative distribution of beads for 2 environmentally realistic concentrations (5 and 100 beads L-1 ) and no substantial egestion of particles within 2 h of depuration. Environ Toxicol Chem 2019;38:99-105. © 2018 SETAC.

A critical perspective on early communications concerning human health aspects of microplastics.

Microplastic research in recent years has shown that small plastic particles are found almost everywhere we look. Besides aquatic and terrestrial environments, this also includes aquatic species intended for human consumption and several studies have reported their prevalence in other food products and beverages. The scientific as well as public debate has therefore increasingly focused on human health implications of microplastic exposure. However, there is a big discrepancy between the magnitude of this debate and actual scientific findings, which have merely shown the presence of microplastics in certain products. While plastics can undoubtedly be hazardous to human health due to toxicity of associated chemicals or as a consequence of particle toxicity, the extent to which microplastics in individual food products and beverages contribute to this is debatable. Considering the enormous use of plastic materials in our everyday lives, microplastics from food products and beverages likely only constitute a minor exposure pathway for plastic particles and associated chemicals to humans. But as this is rarely put into perspective, the recent debate has created a skewed picture of human plastic exposure. We risk pulling the focus away from the root of the problem: the way in which we consume, use and dispose of plastics leading to their widespread presence in our everyday life and in the environment. Therefore we urge for a more careful and balanced discussion which includes these aspects.

Ingestion of micro- and nanoplastics in Daphnia magna - Quantification of body burdens and assessment of feeding rates and reproduction.

Evidence is increasing that micro- and nanoplastic particles can have adverse effects on aquatic organisms. Exposure studies have so far mainly been qualitative since quantitative measurements of particle ingestion are analytically challenging. The aim of this study was therefore to use a quantitative approach for determining ingestion and egestion of micro- and nanoplastics in Daphnia magna and to analyze the influence of particle size, exposure duration and the presence of food. One week old animals were exposed to 2 µm and 100 nm fluorescent polystyrene beads (1 mg/l) for 24 h, followed by a 24 h egestion period in clean medium. During both phases body burdens of particles were determined by measuring the fluorescence intensity in dissolved tissues. Ingestion and egestion were investigated in the absence and presence of food (6.7·105 cells of Raphidocelis subcapitata per ml). Furthermore, feeding rates of daphnids in response to particle exposure were measured as well as effects on reproduction during a 21 days exposure (at 1 mg/l, 0.5 mg/l and 0.1 mg/l) to investigate potential impairments of physiology. Both particle sizes were readily ingested, but the ingested mass of particles was five times higher for the 2 µm particles than for the 100 nm particles. Complete egestion did not occur within 24 h but generally higher amounts of the 2 µm particles were egested. Animal body burdens of particles were strongly reduced in the presence of food. Daphnid feeding rates decreased by 21% in the presence of 100 nm particles, but no effect on reproduction was found despite high body burdens of particles at the end of 21 days exposure. The lower egestion and decreased feeding rates, caused by the 100 nm particles, could indicate that particles in the nanometer size range are potentially more hazardous to D. magna compared to larger particle sizes.

From macro- to microplastics - Analysis of EU regulation along the life cycle of plastic bags.

Plastic pollution and its environmental effects has received global attention the recent years. However, limited attention has so far been directed towards how plastics are regulated in a life cycle perspective and how regulatory gaps can be addressed in order to limit and prevent environmental exposure and hazards of macro- and microplastics. In this paper, we map European regulation taking outset in the life cycle perspective of plastic carrier bags: from plastic bag production to when it enters the environment. Relevant regulatory frameworks, directives and authorities along the life cycle are identified and their role in regulation of plastics is discussed. Most important regulations were identified as: the EU chemical Regulation, the Packaging and Packaging Waste Directive including the amending Directive regarding regulation of the consumption of lightweight plastic carrier bags, the Waste Framework Directive and the Directive on the Landfill of Waste. The main gaps identified relate to lack of clear definitions of categories of polymers, unambitious recycling rates and lack of consideration of macro- and microplastics in key pieces of legislation. We recommend that polymers are categorized according to whether they are polymers with the same monomer constituents (homopolymers) or with different monomer constituents (copolymers) and that polymers are no longer exempt from registration and evaluation under REACH. Plastics should furthermore have the same high level of monitoring and reporting requirements as hazardous waste involving stricter requirements to labelling, recordkeeping, monitoring and control over the whole lifecycle. Finally, we recommend that more ambitious recycle and recovery targets are set across the EU. Regulation of the consumption of lightweight plastic carrier bags should also apply to heavyweight plastic carrier bags. Last, the Marine and Water Framework Directives should specifically address plastic waste affecting water quality.

Microplastics as vectors for environmental contaminants: Exploring sorption, desorption, and transfer to biota.

The occurrence and effects of microplastics (MPs) in the aquatic environment are receiving increasing attention. In addition to their possible direct adverse effects on biota, the potential role of MPs as vectors for hydrophobic organic chemicals (HOCs), compared to natural pathways, is a topic of much debate. It is evident, however, that temporal and spatial variations of MP occurrence do (and will) occur. To further improve the estimations of the role of MPs as vectors for HOC transfer into biota under varying MP concentrations and environmental conditions, it is important to identify and understand the governing processes. Here, we explore HOC sorption to and desorption from MPs and the underlying principles for their interactions. We discuss intrinsic and extrinsic parameters influencing these processes and focus on the importance of the exposure route for diffusive mass transfer. Also, we outline research needed to fill knowledge gaps and improve model-based calculations of MP-facilitated HOC transfer in the environment. Integr Environ Assess Manag 2017;13:488-493. © 2017 SETAC.

Suspended micro-sized PVC particles impair the performance and decrease survival in the Asian green mussel Perna viridis.

Marine bivalves are known to ingest microplastics, but information on the consequences for their physiological performance is limited. To investigate a potential exposure pathway that has not yet been addressed, we mimicked the resuspension of microplastics from the sediment in a laboratory exposure experiment. For this, we exposed the Asian green mussel Perna viridis to 4 concentrations (0mg/l, 21.6mg/l, 216mg/l, 2160mg/l) of suspended polyvinylchloride (PVC) particles (1-50µm) for two 2-hour-time-periods per day. After 44days, mussel filtration and respiration rates as well as byssus production were found to be a negative function of particle concentration. Furthermore, within 91days of exposure, mussel survival declined with increasing PVC abundance. These negative effects presumably go back to prolonged periods of valve closure as a reaction to particle presence. We suggest that microplastics constitute a new seston component that exerts a stress comparable to natural suspended solids.