In recent decades, much attention has been paid to microplastic pollution, and research on microplastics has begun to grow exponentially. However, microplastics research still suffers from the lack of standardized protocols and methods for investigation of microplastics under laboratory conditions. Therefore, in this review, we summarize and critically discuss the results of 715 laboratory studies published on microplastics in the last five years to provide recommendations for future laboratory research. Analysis of the data revealed that the majority of microplastic particles used in laboratory studies are manufactured spheres of polystyrene ranging in size from 1 to 50 µm, that half of the studies did not characterize the particles used, and that a minority of studies used aged particles, investigated leaching of chemicals from microplastics, or used natural particles as a control. There is a large discrepancy between microplastics used in laboratory research and those found in the environment, and many laboratory studies suffer from a lack of environmental relevance and provide incomplete information on the microplastics used. We have summarized and discussed these issues and provided recommendations for future laboratory research on microplastics focusing on (i) microplastic selection, (ii) microplastic characterization, and (iii) test design of laboratory research on microplastics.

Microplastics in the environment occur in different sizes and shapes and are made of various polymers. Therefore, they also considerably differ in their properties and ecotoxicity. However, the majority of microplastics research uses pre-made spherical microplastics, which practically do not exist in the environment. Our work focused on a comprehensive study of six different types of microplastic that were prepared to simulate common microplastics found in the environment. All types of microplastics where chemically and physically characterized using Fourier-transform infrared spectroscopy, thermal analysis, field-emission scanning electron microscopy, optical microscopy and laser diffraction analysis. The specific surface area was determined using the BET method. Furthermore, effects of microplastics and microplastic leachates on a common duckweed (Lemna minor) were evaluated. All tested microplastics did not affect specific growth rate and chlorophyll a content in duckweed, while microplastics with a rough surface and sharp edges caused a significant reduction of duckweed root length. Microplastics made of Bakelite also showed an intensive leaching, which increased their ecotoxicity potential. Natural particles used as a control did not have any negative effect on duckweed. Overall, microplastic particles have significantly different ecotoxicity profiles depending on their physico-chemical properties. Therefore, the testing of environmentally relevant particles and their proper characterization, as well as the testing of microplastic leaching properties, is crucial for understanding of microplastics ecotoxicological potential.

The presence of microplastics (MPs) in the environment has raised many concerns, and therefore approaches and technologies to remove them in situ are of high interest. In this context, we investigated the interactions between polyethylene MPs (fragments with a mean size of 149 ± 75 μm) and an aquatic floating macrophyte Lemna minor in order to assess its potential use for in situ phytoremediation. We first investigated the long-term effects of a high (100 mg/L = 9600 MPs/L), but still environmentally relevant concentration of MPs on L. minor. Subsequently bioadhesion of MPs was studied and the number and strength of MPs adhering to plant biomass were assessed. MPs did not adversely affect various parameters of plants (e.g., specific growth rate, chlorophyll contents, total antioxidant capacity, electron transport system activity, and contents of energy-rich molecules) throughout the duration of the experiment (12 weeks), except for the first week of the experiment, when protein content and total antioxidant capacity were affected. On the other hand, MPs affected the root length of L. minor during the first eight weeks of the experiment, while further exposure resulted in a decrease in the effects, indicating the ability of L. minor to tolerate the presence of MPs for a long period of time. MPs adhered rapidly to the plant biomass and the average percentages of strongly and weakly adhered particles were 6.5% and 20.0%, respectively, of the total MPs applied. In summary, results of this study suggest that L. minor can tolerate hotspot concentrations of MPs and can collect MPs from the water surface. Therefore, phytoremediation using floating plants could be considered as a potential method for in situ removal of MPs from the aquatic environment.

The shape-dependent effects of microplastics have been studied in the context of ingestion but have not been considered in other environmental processes. Therefore, we investigated how the shape of polyethylene microplastics (spheres, fragments, and films) affects interactions with plants, aging, and their adsorption properties. The shape had no effect on the growth rate and chlorophyll content of duckweed Lemna minor, but the fragments strongly adhered to the plant biomass and reduced the root length. The adsorption process of the model organic compound (methylene blue dye) was described by the same kinetic model for all shapes—the experimental data best fit the pseudo-second order model. However, twice as much methylene blue was adsorbed on films as on fragments and spheres. During environmental aging, most biofilm developed on films. The biofilm on spheres contained significantly less photosynthetic microorganisms, but twice as much extracellular polymeric substances (EPS) as on fragments and films. This suggests that the attachment of microorganisms to spherical particles is limited and therefore more intensive production of EPS is required for stable biofilm formation. From the results of this study, it is evident that the shape of microplastics significantly affects not only ecotoxicity but also other environmentally relevant processes.

Microplastics in the aquatic environment serve as a habitat for microbial life, on which they can form biofilms. However, how the development of the biofilm alters the properties of floating microplastics that are at the air-water interface and, therefore, not fully submerged, is not well understood. In this context, an aging experiment was conducted to monitor biofilm formation and changes in physico-chemical properties of low-density polyethylene (floating) microplastics over time. The growth of the biofilm followed the typical bacterial/biofilm growth phases and reached about 30% of the total mass of the microplastics, while the concentration of extracellular polymeric substances within the biofilm remained stable. Presence of chlorophyll a and urease activity indicated presence of photosynthetic microrganisms within the biofilm which was also confirmed by analysis of the biofilm composition. Chemical characterization by FTIR showed the formation of additional functional groups attributed to the formed biofilm, and SEM imaging showed cracks on the surface of the aged microplastics, indicating incipient degradation of the polyethylene. Moreover, the adsorption capacity of the aged particles for metals (Pb(II)) was 52% higher compared to the pristine ones. Aging increased the density and size of the particles; however, it did not lead to the submersion of the aged particles even after 12 weeks of aging, suggesting that additional environmental processes may influence the transport of microplastics from the air-water interface into the water body.