Assessment of the Effects of Environmental Concentrations of Microplastics on the Aquatic Snail Potamopyrgus antipodarum

Microplastics are ubiquitous in aquatic ecosystems. They can be found at the surface, in the water column, and in sediments. Multiple negative impacts of microplastics on aquatic organisms have been reported, with most studies focusing on marine ecosystems. However, the effects of microplastics on freshwater ecosystems have been less studied, with a few studies focusing on benthic invertebrates. In this study, we exposed the New Zealand mud snail Potamopyrgus antipodarum (Gray, 1843) to an environmental range of concentrations of polystyrene microparticles (size range from 0.01 to 514 µm at 100, 500, and 1000 mg microplastics/kg dry weight (dw) of sediment) and two supra-environmental concentrations (2000 and 4000 mg/kg dw sediment). The impacts of the exposure to microplastics on mortality, behavior, and reproduction were assessed at long-term exposure (31 days). Mortality and reproduction were not significantly affected by microplastics. On the contrary, most of the microplastic treatments altered the behavior, causing a significant increase in reaction time compared with controls (0 mg microplastics/kg dw sediment). The highest concentration (4000 mg/kg) did not have an impact on the reaction time over the experimental period compared with controls. To our knowledge, this study is the first to assess the effects of microplastics on the behavior of the aquatic snail P. antipodarum. Our results showed that at environmental concentrations, the behavior of P. antipodarum was the most sensitive variable to the adverse effects of polystyrene microplastics.

Fabrication of Ring-Shaped Deposits of Polystyrene Microparticles Driven by Thermocapillary Mechanism

Fabrication of ring-shaped deposits of microparticles on solid surfaces with the desired length scales and morphology of particle arrangements is of great importance when developing modern optical and electronic resonators, chemical sensors, touch screens, field-emission displays, porous materials, and coatings with various functional properties. However, the controlled formation of ring-shaped patterns scaling from a few millimeters up to centimeters with simultaneous control of particle arrangement at the microscale is one of the most challenging problems in advanced materials science and technology. Here, we report a fabrication approach for ring-shaped structures of microparticles on a glass surface that relied on a local thermal impact produced by the subsurface heater and heat sink. Thermocapillary convection in the liquid covering microparticles in combination with evaporative lithography is responsible for the particle transport and the assembling into the ring-shaped patterns. An advantageous feature of this approach is based on the control of thermocapillary flow direction, achieved by changing the sign of the temperature gradient in the liquid, switching between heating and cooling modes. That allows for changing the particle transfer direction to create the ring-shaped deposits and dynamically tune their size and density distribution. We have studied the influence of the power applied to the heat source/sink and the duration of the applied thermal field on the rate of the ring fabrication, the sizes of the ring and the profile of the particle distribution in the ring. The proposed method is flexible to control simultaneously the centimeter scale and microscale processes of transfer and arrangements of particles and can be applied to the fabrication of ring structures of particles of different nature and shape.

Virgin Polystyrene Microparticles Exposure Leads to Changes in Gills DNA and Physical Condition in the Mediterranean Mussel Mytilus galloprovincialis

The ever-growing concentration of microplastics in the marine environment is leading to a plethora of questions regarding marine organisms’ present and future health status. In this article, the Mediterranean mussel (Mytilus galloprovincialis), a commercial species distributed worldwide, has been exposed to 21 daily doses of polystyrene microparticles (10 µm) at four different concentrations that are environmentally realistic (control: no microplastics, C1: 0.02 mg/L, C2: 0.2 mg/L, and C3: 2 mg/L). The physical status through the condition index, and damages in DNA integrity in gills, through DNA fragmentation, were determined. Results showed a minor effect on DNA integrity but a worse physical status at higher doses. Results could be interpreted as a decrease in mussel feeding activity/filtration rates when exposed to high microplastic concentrations, thus reducing the direct exposure to microplastics in gills. These effects could be happening currently and/or may happen in the near future, threatening populations inhabiting microplastics-polluted environments.

Noxic effects of polystyrene microparticles on murine macrophages and epithelial cells

Microplastic (MP) contamination has been identified as an ecological problem with an increasing impact on everyday life. Yet, possible effects of MP at the cellular level are still poorly understood. Here, the interaction of murine macrophages (J774A.1, ImKC) and epithelial cells (STC-1, BNL CL.2) with well-characterized poly(styrene) MP particles (MPP) of varying sizes (0.2–6.0 µm) was studied. Macrophages are expected to actively engulf particles which could be confirmed in this study, while epithelial cells are found in tissues with direct contact with ingested or inhaled MPP. Here, the epithelial cells from both investigated cell lines did not ingest MPP in significant numbers. Concomitantly, no cytotoxic effects nor any influence on cellular proliferation were observed. Cells from the two macrophage cell lines showed high ingestion of MPP of all sizes, but cytotoxic effects were observed only for one of them (ImKC) and only at MPP concentrations above 250 µg/mL. Indications of cellular stress as well as effects on cell proliferation were observed for cell populations with high particle cell interactions.

Formation of Myoglobin Corona at Polymer Microparticles

Adsorption of myoglobin molecules at negatively charged polystyrene microparticles was studied using the dynamic light scattering (DLS), electrophoresis (LDV) and the solution depletion method involving atomic force microscopy (AFM). The measurements were carried out at pH 3.5 and NaCl concentration of 10−2 and 0.15 M. Initially, the stability of myoglobin solutions and the particle suspensions as a function of pH were determined. Afterward, the formation of myoglobin molecule corona was investigated via the direct electrophoretic mobility measurements, which were converted to the zeta potential. The experimental results were quantitatively interpreted in terms of the general electrokinetic model. This approach yielded the myoglobin corona coverage under in situ conditions. The maximum hard corona coverage was determined using the AFM concentration depletion method. It was equal to 0.9 mg m−2 for the NaCl concentration in the range 0.01 to 0.15 M and pH 3.5. The electrokinetic properties of the corona were investigated using the electrophoretic mobility measurements for a broad pH range. The obtained results confirmed that thorough physicochemical characteristics of myoglobin molecules can be acquired using nM amounts of the protein. It was also argued that this method can be used for performing electrokinetic characteristics of other proteins such as the SARS-Cov-2 spike protein exhibiting, analogously to myoglobin, a positive charge at acidic pHs.

Microfluidic Electric Egg-Laying Assay and Application to In-vivo Toxicity Screening of Microplastics using C. elegans

Environmental pollutants like microplastics are posing health concerns on aquatic animals and the ecosystem. Microplastic toxicity studies using C. elegans as a model are evolving but methodologically hindered from obtaining statistically strong data sets, detecting toxicity effects based on microplastics uptake, and correlating physiological and behavioural effects at an individual-worm level. In this paper, we report a novel microfluidic electric egg-laying assay for phenotypical assessment of multiple worms in parallel. The effects of glucose and polystyrene microplastics at various concentrations on the worms’ electric egg-laying, length, diameter, and length contraction during exposure to electric signal were studied. The device contained eight parallel worm-dwelling microchannels called electric traps, with equivalent electrical fields, in which the worms were electrically stimulated for egg deposition and fluorescently imaged for assessment of neuronal and microplastic uptake expression. A new bidirectional stimulation technique was developed, and the device design was optimized to achieve a testing efficiency of 91.25%. Exposure of worms to 100mM glucose resulted in a significant reduction in their egg-laying and size. The effects of 1μm polystyrene microparticles at concentrations of 100 and 1000 mg/L on the electric egg-laying behaviour, size, and neurodegeneration of N2 and NW1229 (expressing GFP pan-neuronally) worms were also studied. Of the two concentrations, 1000 mg/L caused severe egg-laying deficiency and growth retardation as well as neurodegeneration. Additionally, using single-worm level phenotyping, we noticed intra-population variability in microplastics uptake and correlation with the above physiological and behavioural phenotypes, which was hidden in the population-averaged results. Taken together, these results suggest the appropriateness of our microfluidic assay for toxicological studies and for assessing the phenotypical heterogeneity in response to microplastics.