In vitro evaluation of cytotoxic and genotoxic effects of Di(2-ethylhexyl)-phthalate (DEHP) on European sea bass (Dicentrarchus labrax) embryonic cell line

Marine litter is extensively distributed in the marine environment, and plastic debris, of which litter is mostly composed, can be a major source of pollutants. Among them, Di(2-ethylhexyl)-phthalate (DEHP) is the most abundantly used plastic additive, and it has been reported to affect biochemical processes both in humans and wildlife; however, studies on its toxicological effects on marine organisms are still scarce. In this survey, we studied the cytotoxic, genotoxic, and mutagenic effects of DEHP in European sea bass embryonic cell line (DLEC) by applying specific in vitro tests. Results showed a significant decrease in cell viability starting at 0.01 mM of DEHP after 24 h together with a significant increase in apoptosis and necrosis, morphological changes and cell detachment. Consistently, we detected a moderate increase in DNA strand breaks from 0.02 mM, and a dose-dependent increase in of micronucleus frequency from 0.01 mM, accompanied by a significant inhibition of cell proliferation, which suggested a possible aneugenic effect of this phthalate. Our results demonstrate that in vitro exposure to DEHP had a dose-dependent cytotoxic and genotoxic effects in DLEC cell line, encouraging further investigation into its effects in in vivo and/or ex vivo cell systems of marine organisms.

Molino C., Filippi S., Stoppiello G. A. and al., Toxicology in Vitro, Volume 56, April 2019, Pages 118-125

The article


Cellular responses of Pacific oyster (Crassostrea gigas) gametes exposed in vitro to polystyrene nanoparticles

While the detection and quantification of nano-sized plastic in the environment remains a challenge, the growing number of polymer applications mean that we can expect an increase in the release of nanoplastics into the environment by indirect outputs. Today, very little is known about the impact of nano-sized plastics on marine organisms. Thus, the objective of this study was to investigate the toxicity of polystyrene nanoplastics (NPs) on oyster (Crassostrea gigas) gametes. Spermatozoa and oocytes were exposed to four NPs concentrations ranging from 0.1 to 100 mg L−1 for 1, 3 and 5 h. NPs coated with carboxylic (PS-COOH) and amine groups (PS-NH2) were used to determine how surface properties influence the effects of nanoplastics. Results demonstrated the adhesion of NPs to oyster spermatozoa and oocytes as suggested by the increase of relative cell size and complexity measured by flow-cytometry and confirmed by microscopy observations. A significant increase of ROS production was observed in sperm cells upon exposure to 100 mg L−1 PS-COOH, but was not observed with PS-NH2, suggesting a differential effect according to the NP-associated functional group. Altogether, these results demonstrate that the effects of NPs occur rapidly, are complex and are possibly associated with the cellular eco-corona, which could modify NPs behaviour and toxicity.

Gonzalez-Fernandez C., Tallec K, Le Goïc N. and al., Chemosphere, Volume 208, October 2018, Pages 764-772

The article

Do microplastics affect marine ecosystem productivity?

Marine and coastal ecosystems are among the largest contributors to the Earth’s productivity. Experimental studies have shown negative impacts of microplastics on individual algae or zooplankton organisms. Consequently, primary and secondary productivity may be negatively affected as well. In this study we attempted to estimate the impacts on productivity at ecosystem level based on reported laboratory findings with a modelling approach, using our biogeochemical model for the North Sea (Delft3D-GEM). Although the model predicted that microplastics do not affect the total primary or secondary production of the North Sea as a whole, the spatial patterns of secondary production were altered, showing local changes of ±10%. However, relevant field data on microplastics are scarce, and strong assumptions were required to include the plastic concentrations and their impacts under field conditions into the model. These assumptions reveal the main knowledge gaps that have to be resolved to improve the first estimate above.

Troost T. A., Desclaux T., Leslie H. A. and al., Marine Pollution Bulletin, Volume 135, October 2018, Pages 17-29

The article

Nanoplastics impaired oyster free living stages, gametes and embryos

In the marine environment, most bivalve species base their reproduction on external fertilization. Hence, gametes and young stages face many threats, including exposure to plastic wastes which represent more than 80% of the debris in the oceans. Recently, evidence has been produced on the presence of nanoplastics in oceans, thus motivating new studies of their impacts on marine life. Because no information is available about their environmental concentrations, we performed dose-response exposure experiments with polystyrene particles to assess the extent of micro/nanoplastic toxicity. Effects of polystyrene with different sizes and functionalizations (plain 2-μm, 500-nm and 50-nm; COOH-50 nm and NH2-50 nm) were assessed on three key reproductive steps (fertilization, embryogenesis and metamorphosis) of Pacific oysters (Crassostrea gigas). Nanoplastics induced a significant decrease in fertilization success and in embryo-larval development with numerous malformations up to total developmental arrest. The NH2-50 beads had the strongest toxicity to both gametes (EC50 = 4.9 μg/mL) and embryos (EC50 = 0.15 μg/mL), showing functionalization-dependent toxicity. No effects of plain microplastics were recorded. These results highlight that exposures to nanoplastics may have deleterious effects on planktonic stages of oysters, presumably interacting with biological membranes and causing cyto/genotoxicity with potentially drastic consequences for their reproductive success.

Tallec K., Huvet A., Di Poi C. and al., Environmental Pollution, Volume 242, Part B, November 2018, Pages 1226-1235

The article

Microplastic pollution in commercial salt for human consumption: A review

Microplastics (MPs) are plastic particles with less than 5 mm in size that are considered global environmental pollutants. The MPs present in the environment result from the successive breakdown of larger plastic pieces or from the direct input of micro- and nano-sized particles used in various industries and products available to consumers. Such MPs have been found in several wild species and other natural resources, including some consumed as food by humans, with possible adverse effects on ecosystem and human health. The central aim of this work was to review the published literature regarding the contamination of sea commercial salts (sea and terrestrial origins) and its possible impacts on human health. Moreover, to lead to a comprehensive understanding of the paradigm, a short introduction and revision of the environmental contamination by MPs and its effects are included. MPs have been found in commercial salts from 128 brands, from 38 different countries spanning over five continents. The concentration of MPs found in the samples analysed is lower than the concentrations of MPs reported in other resources, such as blue mussels. However, as commercial salts are used every day and by all humans, they constitute a long-term exposure route for the general population in addition to others (e.g., animals consumed as food by humans, water, air). Therefore, commercial salts contaminated with MPs may contribute to the potential long-term adverse effects resulting from human exposure to these particles.

Peixoto D., Pinheiro C., Amorim J. and al., Estuarine, Coastal and Shelf Science, Volume 219, 5 April 2019, Pages 161-168

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Effects of microplastics on microalgae populations: A critical review

Microplastics are persistent contaminants accumulating in the environment. Aquatic ecosystems have been studied worldwide, revealing ubiquitous contamination with microplastics. Microalgae, one of the most important primary producers in aquatic ecosystems, could suffer from microplastic contamination, leading to larger impacts on aquatic food webs. Nonetheless, little is known about the toxic effects of microplastics on microalgae populations. Thus, the objective of this review was to identify these effects and the impacts of microplastics on microalgae populations based on currently available literature, also identifying knowledge gaps. Even though microplastics seem to have limited effects on parameters such as growth, chlorophyll content, photosynthesis activity and reactive oxygen species (ROS), current environmental concentrations are not expected to induce toxicity. Even so, microplastics could disrupt population regulation mechanisms, by reducing the availability or absorption of nutrients (bottom-up) or reducing the population of predator species (top-down). Microplastics’ properties can also influence the effects on microalgae, with smaller sizes and positive surface charges having higher toxicity. Therefore, more research is needed to better understand the effects of microplastics on microalgae, such as adaptation strategies, effects on population dynamics and microplastics properties influencing toxicity.

Correia Prata J., da Costa J. P., Lopes I. and al., Science of The Total Environment, Volume 665, 15 May 2019, Pages 400-405

The article

Combined effect of polystyrene plastics and triphenyltin chloride on the green algae Chlorella pyrenoidosa

The combined effect of polystyrene (PS) particles and triphenyltin chloride (TPTCl) to the green algae Chlorella pyrenoidosa was studied. The 96 h IC50 of TPTCl to the green algae C. pyrenoidosa was 30.64 μg/L. The toxicity of PS particles to C. pyrenoidosa was size-dependent, with the 96 h IC50 at 9.10 mg/L for 0.55 μm PS but no toxicity observed for 5.0 μm PS. The exposure to 0.55 μm PS led to damage on structure of algal cells, which could in turn cause inhibition on photosynthesis and population growth of the green algae. TPTCl concentrations in test medium were lowered by 15–19% at presence of 0.55 μm PS particles, indicating a reduced bioavailability of TPTCl. In spite of this reduced bioavailability, the presence of PS increased the toxicity of TPTCl, which might be attributed to facilitated uptake of TPTCl by the green algae after the damage of cell structure. The overall results of the present study provided important information on the effect of PS on the bioavailability and toxicity of TPTCl to phytoplankton species.

Yi, X., Chi, T., Li, Z. et al. Environ Sci Pollut Res (2019)

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