Toxic effects of pristine and aged polystyrene and their leachate on marine microalgae Skeletonema costatum.

The acute toxic effects of pristine and aged polystyrene (P-PS and A-PS) and their leaching solutions (L-PS) on microalgae Skeletonema costatum were investigated by measuring algal density and growth inhibition rate (IR), chlorophyll concentration and photosynthetic efficiency (Fv/Fm) over 96 h. Total protein (TP), superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) were measured to analyze the oxidative damage to microalgae by microplastics and their leachates. Hydrodynamic diameter of microplastics in seawater, FITR and SEM images were used to study the changes of polystyrene during aging. The interaction of algae cell with microplastics and the cellular ultrastructure changes of cells were analyzed combined with electron microscopy for a comprehensive and systematic understanding on the mechanisms of microplastic toxicity to microalgae. Both high concentration and small size of PS had significant inhibitory effect on the growth of microalgae, and the inhibitory effect was greater with increasing exposure time. The inhibition effect of aged microplastics was more obvious, which was speculated to be caused by the synergistic effect of aged PS itself and leaching solution. The negative effect of leaching solution on microalgae was due to the release of some additives during the aging process. The content of MDA reached the highest value of 54.41 nmol/mgprot in 1.0 µm 50 mg/L A-PS treatment group, and A-PS were found to be more prone to heterogeneous aggregation with algae cells by SEM.

Is hydrodynamic diameter the decisive factor? - Comparison of the toxic mechanism of nSiO2 and mPS on marine microalgae Heterosigma akashiwo.

To investigate the toxic mechanism of SiO2 nanoparticles (nSiO2) and polystyrene microplastics (mPS) on microalgae Heterosigma akashiwo, growth inhibition tests were carried out. The growth and biological responses of the algae exposed to nSiO2 (0.5, 1, 1.5, 2, 5, 10 and 30 mg L-1) and mPS (1, 2, 5, 10, 30 and 75 mg L-1) were explored in f/2 media for 96 h. It was found that the hydrodynamic diameter of the particles seems to be one of the more important factors to influence the algae. nSiO2 and mPS with similar hydrodynamic diameters have the similar toxic mechanism on H. akashiwo, and the effects were dose- and time-dependent. High concentrations of micro-/nano-particles (MNPs) could inhibit the growth of algal cells, however, low concentrations of MNPs did not restrict or even promoted the growth of algae, known as "Hormesis" phenomenon. The 96 h-EC20 values of nSiO2 and mPS on H. akashiwo were 2.69 and 10.07 mg L-1, respectively, and chlorophyll fluorescence parameters indicated that the microalgal photosynthetic system were inhibited. The hydrophilic surface of nSiO2 increased the likelihood of nSiO2 binding to the hydrophilic functional group of microalgae, which may account for the slightly stronger toxic effect of nSiO2 than mPS. The algae continued to produce reactive oxygen species (ROS) under stress conditions. Total protein (TP) levels reduced, and superoxide dismutase (SOD) and catalase (CAT) levels increased to maintain ROS levels in the cells. The decrease in adenosine triphosphate (ATPase) indicated an impact on cellular energy metabolism. Cell membrane damage, cytoplasm and organelle efflux under stress were confirmed by scanning and transmission electron microscopy (SEM and TEM) images. This study contributes to the understanding of the size effect of MNPs on the growth of marine microalgae.

Microplastic-induced apoptosis and metabolism responses in marine Dinoflagellate, Karenia mikimotoi.

Microplastics (MPs) occur widely in marine environments, and disturb the balance of aquatic ecosystems. In this study, programmed cell apoptosis in marine dinoflagellate, Karenia mikimotoi exposed to 10 mg L-1 micro/nanoplastics (MPs/NPs; polystyrene and polymethyl methacrylate) for 72 h was assessed. Prior to the toxicity assay, MPs/NPs were dialyzed to remove possible additives. Cell viability, membrane integrity, cell apoptosis, and total DNA concentration were measured to assess programmed cell apoptosis in K. mikimotoi following exposure to MPs/NPs. A transcriptome analysis was used to explore the potential toxic mechanism of MPs to K. mikimotoi. Programmed cell apoptosis was related to the size of MPs/NPs, and NPs could more easily impair cell viability, and reduced cell membrane integrity and DNA concentration. NP particles caused continuous apoptosis of K. mikimotoi compared to MP particles. Size had the greatest effect on toxicity in K. mikimotoi. In conclusion, the results evidenced that both MPs and NPs have a negative impact on the marine dinoflagellate, K. mikimotoi. However, NPs were more harmful to K mikimotoi than MPs, highlighting the potential ecological problems associated with exposure to NPs.

The effects and mechanisms of polystyrene and polymethyl methacrylate with different sizes and concentrations on Gymnodinium aeruginosum.

In this study, Gymnodinium aeruginosum was exposed to polystyrene (PS) and polymethyl methacrylate (PMMA) of three particle sizes (0.1 µm, 1.0 µm and 100 µm) and two concentrations (10 mg/L and 75 mg/L) for 96 h. The density of algae cells, the endpoints that reactive oxygen species (ROS), total protein (TP), malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT), scanning and transmission electron microscopy (SEM and TEM) were used to explore the toxicity mechanism to the microalgae. At a concentration of 75 mg/L, the 96 h inhibition ratios (IR) with particle sizes of 0.1 µm, 1.0 µm and 100 µm on G. aeruginosum were 55.9%, 63.7% and 6.0% for PS, respectively, and 3.0%, 4.1% and -0.6% for PMMA, respectively. The most significant changes in ROS, TP, MDA, SOD and CAT were observed at 75 mg/L 1.0 µm of PS when treated for 96 h. When exposed to nanoplastics (NPs) and microplastics (MPs), the algae cells were damaged, and the antioxidant system was activated. Extracellular polymeric substance (EPS) could help to detoxify the algae. In general, PS was more toxic than PMMA. The toxicity of small MNPs (0.1 µm and 1.0 µm) was related to the concentrations, while large MNPs (100 µm) did not.