Quality Criteria for the Analysis of Microplastic in Biota Samples: A Critical Review.

Data on ingestion of microplastics by marine biota are quintessential for monitoring and risk assessment of microplastics in the environment. Current studies, however, portray a wide spread in results on the occurrence of microplastic ingestion, highlighting a lack of comparability of results, which might be attributed to a lack of standardization of methods. We critically review and evaluate recent microplastic ingestion studies in aquatic biota, propose a quality assessment method for such studies, and apply the assessment method to the reviewed studies. The quality assessment method uses ten criteria: sampling method and strategy, sample size, sample processing and storage, laboratory preparation, clean air conditions, negative controls, positive controls, target component, sample (pre)treatment, and polymer identification. The results of this quality assessment show a dire need for stricter quality assurance in microplastic ingestion studies. On average, studies score 8.0 out of 20 points for "completeness of information" and 0 for "reliability". Alongside the assessment method, a standardized protocol for detecting microplastic in biota samples incorporating these criteria is provided.

Risks of Plastic Debris: Unravelling Fact, Opinion, Perception, and Belief.

Researcher and media alarms have caused plastic debris to be perceived as a major threat to humans and animals. However, although the waste of plastic in the environment is clearly undesirable for aesthetic and economic reasons, the actual environmental risks of different plastics and their associated chemicals remain largely unknown. Here we show how a systematic assessment of adverse outcome pathways based on ecologically relevant metrics for exposure and effect can bring risk assessment within reach. Results of such an assessment will help to respond to the current public worry in a balanced way and allow policy makers to take measures for scientifically sound reasons.

Export of microplastics from land to sea. A modelling approach.

Quantifying the transport of plastic debris from river to sea is crucial for assessing the risks of plastic debris to human health and the environment. We present a global modelling approach to analyse the composition and quantity of point-source microplastic fluxes from European rivers to the sea. The model accounts for different types and sources of microplastics entering river systems via point sources. We combine information on these sources with information on sewage management and plastic retention during river transport for the largest European rivers. Sources of microplastics include personal care products, laundry, household dust and tyre and road wear particles (TRWP). Most of the modelled microplastics exported by rivers to seas are synthetic polymers from TRWP (42%) and plastic-based textiles abraded during laundry (29%). Smaller sources are synthetic polymers and plastic fibres in household dust (19%) and microbeads in personal care products (10%). Microplastic export differs largely among European rivers, as a result of differences in socio-economic development and technological status of sewage treatment facilities. About two-thirds of the microplastics modelled in this study flow into the Mediterranean and Black Sea. This can be explained by the relatively low microplastic removal efficiency of sewage treatment plants in the river basins draining into these two seas. Sewage treatment is generally more efficient in river basins draining into the North Sea, the Baltic Sea and the Atlantic Ocean. We use our model to explore future trends up to the year 2050. Our scenarios indicate that in the future river export of microplastics may increase in some river basins, but decrease in others. Remarkably, for many basins we calculate a reduction in river export of microplastics from point-sources, mainly due to an anticipated improvement in sewage treatment.

Aging of microplastics promotes their ingestion by marine zooplankton.

Microplastics (<5 mm) are ubiquitous in the marine environment and are ingested by zooplankton with possible negative effects on survival, feeding, and fecundity. The majority of laboratory studies has used new and pristine microplastics to test their impacts, while aging processes such as weathering and biofouling alter the characteristics of plastic particles in the marine environment. We investigated zooplankton ingestion of polystyrene beads (15 and 30 µm) and fragments (≤30 µm), and tested the hypothesis that microplastics previously exposed to marine conditions (aged) are ingested at higher rates than pristine microplastics. Polystyrene beads were aged by soaking in natural local seawater for three weeks. Three zooplankton taxa ingested microplastics, excluding the copepod Pseudocalanus spp., but the proportions of individuals ingesting plastic and the number of particles ingested were taxon and life stage specific and dependent on plastic size. All stages of Calanus finmarchicus ingested polystyrene fragments. Aged microbeads were preferred over pristine ones by females of Acartia longiremis as well as juvenile copepodites CV and adults of Calanus finmarchicus. The preference for aged microplastics may be attributed to the formation of a biofilm. Such a coating, made up of natural microbes, may contain similar prey as the copepods feed on in the water column and secrete chemical exudates that aid chemodetection and thus increase the attractiveness of the particles as food items. Much of the ingested plastic was, however, egested within a short time period (2-4 h) and the survival of adult Calanus females was not affected in an 11-day exposure. Negative effects of microplastics ingestion were thus limited. Our findings emphasize, however, that aging plays an important role in the transformation of microplastics at sea and ingestion by grazers, and should thus be considered in future microplastics ingestion studies and estimates of microplastics transfer into the marine food web.

The Effect of Microplastic on the Uptake of Chemicals by the Lugworm Arenicola marina (L.) under Environmentally Relevant Exposure Conditions.

It has been hypothesized that ingestion of microplastic increases exposure of aquatic organisms to hydrophobic contaminants. To date, most laboratory studies investigated chemical transfer from ingested microplastic without taking other exposure pathways into account. Therefore, we studied the effect of polyethylene (PE) microplastic in sediment on PCB uptake by Arenicola marina as a model species, quantifying uptake fluxes from all natural exposure pathways. PCB concentrations in sediment, biota lipids (Clip) and porewater measured with passive samplers were used to derive lipid-normalized bioaccumulation metrics Clip, Biota sediment accumulation factor (BSAF), Bioaccumulation factor (BAF) and the Biota plastic accumulation factor (BPAF). Small effects of PE addition were detected suggesting slightly increased or decreased bioaccumulation. However, the differences decreased in magnitude dependent on the metric used to assess bioaccumulation, in the order: Clip > BSAF > BPAF > BAF, and were nonsignificant for BAF. The fact that BAF, that is, normalization of Clip on porewater concentration, largely removed all effects of PE, shows that PE did not act as a measurable vector of PCBs. Biodynamic model analysis confirmed that PE ingestion contributed marginally to bioaccumulation. This work confirmed model-based predictions on the limited relevance of microplastic for bioaccumulation under environmentally realistic conditions, and illustrated the importance of assessing exposure through all media in microplastic bioaccumulation studies.

Detection of low numbers of microplastics in North Sea fish using strict quality assurance criteria.

We investigated 400 individual fish of four North Sea species: Atlantic Herring, Sprat, Common Dab, and Whiting on ingestion of >20µm microplastic. Strict quality assurance criteria were followed in order to control contamination during the study. Two plastic particles were found in only 1 (a Sprat) out of 400 individuals (0.25%, with a 95% confidence interval of 0.09-1.1%). The particles were identified to consist of polymethylmethacrylate (PMMA) through FTIR spectroscopy. No contamination occurred during the study, showing the method applied to be suitable for microplastic ingestion studies in biota. We discuss the low particle count for North Sea fish with those in other studies and suggest a relation between reported particle count and degree of quality assurance applied. Microplastic ingestion by fish may be less common than thought initially, with low incidence shown in this study, and other studies adhering to strict quality assurance criteria.

Fate of nano- and microplastic in freshwater systems: A modeling study.

Riverine transport to the marine environment is an important pathway for microplastic. However, information on fate and transport of nano- and microplastic in freshwater systems is lacking. Here we present scenario studies on the fate and transport of nano-to millimetre sized spherical particles like microbeads (100 nm-10 mm) with a state of the art spatiotemporally resolved hydrological model. The model accounts for advective transport, homo- and heteroaggregation, sedimentation-resuspension, polymer degradation, presence of biofilm and burial. Literature data were used to parameterize the model and additionally the attachment efficiency for heteroaggregation was determined experimentally. The attachment efficiency ranged from 0.004 to 0.2 for 70 nm and 1050 nm polystyrene particles aggregating with kaolin or bentonite clays in natural freshwater. Modeled effects of polymer density (1-1.5 kg/L) and biofilm formation were not large, due to the fact that variations in polymer density are largely overwhelmed by excess mass of suspended solids that form heteroaggregates with microplastic. Particle size had a dramatic effect on the modeled fate and retention of microplastic and on the positioning of the accumulation hot spots in the sediment along the river. Remarkably, retention was lowest (18-25%) for intermediate sized particles of about 5 µm, which implies that the smaller submicron particles as well as larger micro- and millimetre sized plastic are preferentially retained. Our results suggest that river hydrodynamics affect microplastic size distributions with profound implications for emissions to marine systems.

Incorporation of microplastics from litter into burrows of Lumbricus terrestris.

Pollution caused by plastic debris is an urgent environmental problem. Here, we assessed the effects of microplastics in the soil surface litter on the formation and characterization of burrows built by the anecic earthworm Lumbricus terrestris in soil and quantified the amount of microplastics that was transported and deposited in L. terrestris burrows. Worms were exposed to soil surface litter treatments containing microplastics (Low Density Polyethylene) for 2 weeks at concentrations of 0%, 7%, 28%, 45% and 60%. The latter representing environmentally realistic concentrations found in hot spot soil locations. There were significantly more burrows found when soil was exposed to the surface treatment composed of 7% microplastics than in all other treatments. The highest amount of organic matter in the walls of the burrows was observed after using the treatments containing 28 and 45% microplastics. The highest microplastic bioturbation efficiency ratio (total microplastics (mg) in burrow walls/initial total surface litter microplastics (mg)) was found using the concentration of 7% microplastics, where L. terrestris introduced 73.5% of the surface microplastics into the burrow walls. The highest burrow wall microplastic content per unit weight of soil (11.8 ± 4.8 g kg-1) was found using a concentration of 60% microplastics. L. terrestris was responsible for size-selective downward transport when exposed to concentrations of 7, 28 and 45% microplastics in the surface litter, as the fraction ≤50 µm microplastics in burrow walls increased by 65% compared to this fraction in the original surface litter plastic. We conclude that the high biogenic incorporation rate of the small-fraction microplastics from surface litter into burrow walls causes a risk of leaching through preferential flow into groundwater bodies. Furthermore, this leaching may have implications for the subsequent availability of microplastics to terrestrial organisms or for the transport of plastic-associated organic contaminants in soil.

Microplastics in the Terrestrial Ecosystem: Implications for Lumbricus terrestris (Oligochaeta, Lumbricidae).

Plastic debris is widespread in the environment, but information on the effects of microplastics on terrestrial fauna is completely lacking. Here, we studied the survival and fitness of the earthworm Lumbricus terrestris (Oligochaeta, Lumbricidae) exposed to microplastics (Polyethylene, <150 µm) in litter at concentrations of 7, 28, 45, and 60% dry weight, percentages that, after bioturbation, translate to 0.2 to 1.2% in bulk soil. Mortality after 60 days was higher at 28, 45, and 60% of microplastics in the litter than at 7% w/w and in the control (0%). Growth rate was significantly reduced at 28, 45, and 60% w/w microplastics, compared to the 7% and control treatments. Due to the digestion of ingested organic matter, microplastic was concentrated in cast, especially at the lowest dose (i.e., 7% in litter) because that dose had the highest proportion of digestible organic matter. Whereas 50 percent of the microplastics had a size of <50 µm in the original litter, 90 percent of the microplastics in the casts was <50 µm in all treatments, which suggests size-selective egestion by the earthworms. These concentration-transport and size-selection mechanisms may have important implications for fate and risk of microplastic in terrestrial ecosystems.

Nanoplastic affects growth of S. obliquus and reproduction of D. magna.

The amount of nano- and microplastic in the aquatic environment rises due to the industrial production of plastic and the degradation of plastic into smaller particles. Concerns have been raised about their incorporation into food webs. Little is known about the fate and effects of nanoplastic, especially for the freshwater environment. In this study, effects of nano-polystyrene (nano-PS) on the growth and photosynthesis of the green alga Scenedesmus obliquus and the growth, mortality, neonate production, and malformations of the zooplankter Daphnia magna were assessed. Nano-PS reduced population growth and reduced chlorophyll concentrations in the algae. Exposed Daphnia showed a reduced body size and severe alterations in reproduction. Numbers and body size of neonates were lower, while the number of neonate malformations among neonates rose to 68% of the individuals. These effects of nano-PS were observed between 0.22 and 103 mg nano-PS/L. Malformations occurred from 30 mg of nano-PS/L onward. Such plastic concentrations are much higher than presently reported for marine waters as well as freshwater, but may eventually occur in sediment pore waters. As far as we know, these results are the first to show that direct life history shifts in algae and Daphnia populations may occur as a result of exposure to nanoplastic.

Leaching of plastic additives to marine organisms.

It is often assumed that ingestion of microplastics by aquatic species leads to increased exposure to plastic additives. However, experimental data or model based evidence is lacking. Here we assess the potential of leaching of nonylphenol (NP) and bisphenol A (BPA) in the intestinal tracts of Arenicola marina (lugworm) and Gadus morhua (North Sea cod). We use a biodynamic model that allows calculations of the relative contribution of plastic ingestion to total exposure of aquatic species to chemicals residing in the ingested plastic. Uncertainty in the most crucial parameters is accounted for by probabilistic modeling. Our conservative analysis shows that plastic ingestion by the lugworm yields NP and BPA concentrations that stay below the lower ends of global NP and BPA concentration ranges, and therefore are not likely to constitute a relevant exposure pathway. For cod, plastic ingestion appears to be a negligible pathway for exposure to NP and BPA.

Plastic as a carrier of POPs to aquatic organisms: a model analysis.

It has been hypothesized that persistent organic pollutants (POPs) in microplastic may pose a risk to aquatic organisms. Here we develop and analyze a conceptual model that simulates the effects of plastic on bioaccumulation of POPs. The model accounts for dilution of exposure concentration by sorption of POPs to plastic (POP "dilution"), increased bioaccumulation by ingestion of plastic-containing POPs ("carrier"), and decreased bioaccumulation by ingestion of clean plastic ("cleaning"). The model is parametrized for the lugworm Arenicola marina and evaluated against recently published bioaccumulation data for this species from laboratory bioassays with polystyrene microplastic. Further scenarios include polyethylene microplastic, nanosized plastic, and open marine systems. Model analysis shows that plastic with low affinity for POPs such as polystyrene will have a marginal decreasing effect on bioaccumulation, governed by dilution. For stronger sorbents such as polyethylene, the dilution, carrier, and cleaning mechanism are more substantial. In closed laboratory bioassay systems, dilution and cleaning dominate, leading to decreased bioaccumulation. Also in open marine systems a decrease is predicted due to a cleaning mechanism that counteracts biomagnification. However, the differences are considered too small to be relevant from a risk assessment perspective.

Effects of microplastic on fitness and PCB bioaccumulation by the lugworm Arenicola marina (L.).

It has been speculated that marine microplastics may cause negative effects on benthic marine organisms and increase bioaccumulation of persistent organic pollutants (POPs). Here, we provide the first controlled study of plastic effects on benthic organisms including transfer of POPs. The effects of polystyrene (PS) microplastic on survival, activity, and bodyweight, as well as the transfer of 19 polychlorinated biphenyls (PCBs), were assessed in bioassays with Arenicola marina (L.). PS was pre-equilibrated in natively contaminated sediment. A positive relation was observed between microplastic concentration in the sediment and both uptake of plastic particles and weight loss by A. marina. Furthermore, a reduction in feeding activity was observed at a PS dose of 7.4% dry weight. A low PS dose of 0.074% increased bioaccumulation of PCBs by a factor of 1.1-3.6, an effect that was significant for ΣPCBs and several individual congeners. At higher doses, bioaccumulation decreased compared to the low dose, which however, was only significant for PCB105. PS had statistically significant effects on the organisms' fitness and bioaccumulation, but the magnitude of the effects was not high. This may be different for sites with different plastic concentrations, or plastics with a higher affinity for POPs.