Do tidal fluctuations affect microplastics distribution and composition in coastal waters?

The hydro-meteorological conditions in marine environments are recognized to have a major impact on the transport and dispersion of microplastics (MP), although their precise effects remain poorly understood. This study investigates the effects of tidal fluctuations on MP abundance and composition in a megatidal coastal water. Waters samples were collected every ninety minutes over the course of two complete tidal cycles - one during spring tide and another during neap tide. There were no significant disparities in term of abondance, size, and composition of MPs between the samples collected during the two tidal cycles. Nevertheless, MP abundance and characteristics (morphology, size and polymer types) can be influenced over the course of a complete tidal cycle due to the impact of tidal currents and water height. This study highlights the need to consider the fluctuations of the tidal cycle when planning in-situ surveys to better assess MP pollution in coastal environments.

Microplastic inputs to the Mediterranean Sea during wet and dry seasons: The case of two Lebanese coastal outlets.

Few studies have highlighted the impact of urbanization and meteorological events on the quantity of microplastics (MPs) discharged into the sea through rivers. To evaluate this issue in the Mediterranean Basin, surface water samples were collected from two more or less urbanized Lebanese Rivers: the Nahr Ibrahim (S1) and the Nahr Antelias (S2), during dry and wet periods. A significant higher abundance of 14.02 ± 9.8 particles/L was reported in the most industrialized river S2 compared to 1.73 ± 1.38 particles/L at S1. A correlation was found between particle contamination and the season at each site. Our results indicate that the MP concentrations were highest on the first sampling day of the wet season and tended to decrease progressively with increasing cumulative precipitation. Some polymers were identified only during one season. Meteorological events should be taken more specifically into account in order to define the influx of plastic pollution into coastal waters more accurately.

Vertical distribution of microplastics in a river water column using an innovative sampling method.

Due to limitations of sampling methods, subsurface water is usually a less well-investigated compartment of the water column when scientists assess microplastic contamination. In this study, microplastic (MP) contamination was assessed in a freshwater river both in surface and subsurface using an innovative sampling method. Microplastic contamination in the lower part of the water column, i.e., near-bottom water and in sediments, was also studied. Three sampling campaigns were carried out during different weather conditions: stormy, rainy, and dry in order to observe their influence on the microplastics vertical distribution. No significant difference was observed between the abundance and types of MPs in surface and subsurface water. The proportion of polymer with theoretical density < 1 (polypropylene d = 0.9, polyethylene d = 0.91-0.95) and polystyrene (d = 0.1-1.06) in the surface and subsurface samples was 73.5%, and this proportion drops to 40.8% for the samples located in the near-bottom water and the sediments. Our results indicate that the MP concentration of the different compartments analyzed can be significantly influenced by rainfall during and prior to the sampling day. This study highlights that in shallow rivers, surface water sampling is representative of the water column MP contamination, but that sampling without taking environmental conditions into account may lead to erroneous estimation of MPs concentration and flux entering the marine environment.

Additives in polypropylene and polylactic acid food packaging: Chemical analysis and bioassays provide complementary tools for risk assessment.

Plastic food packaging represents 40 % of the plastic production worldwide and belongs to the 10 most commonly found items in aquatic environments. They are characterized by high additives contents with >4000 formulations available on the market. Thus they can release their constitutive chemicals (i.e. additives) into the surrounding environment, contributing to chemical pollution in aquatic systems and to contamination of marine organism up to the point of questioning the health of the consumer. In this context, the chemical and toxicological profiles of two types of polypropylene (PP) and polylactic acid (PLA) food packaging were investigated, using in vitro bioassays and target gas chromatography mass spectrometry analyses. Plastic additives quantification was performed both on the raw materials, and on the material leachates after 5 days of lixiviation in filtered natural seawater. The results showed that all samples (raw materials and leachates) contained additive compounds (e.g. phthalates plasticizers, phosphorous flame retardants, antioxidants and UV-stabilizers). Differences in the number and concentration of additives between polymers and suppliers were also pointed out, indicating that the chemical signature cannot be generalized to a polymer and is rather product dependent. Nevertheless, no significant toxic effects was observed upon exposure to the leachates in two short-term bioassays targeting baseline toxicity (Microtox® test) and Pacific oyster Crassostrea gigas fertilization success and embryo-larval development. Overall, this study demonstrates that both petrochemical and bio-based food containers contain harmful additives and that it is not possible to predict material toxicity solely based on chemical analysis. Additionally, it highlights the complexity to assess and comprehend the additive content of plastic packaging due to the variability of their composition, suggesting that more transparency in polymer formulations is required to properly address the risk associated with such materials during their use and end of life.

An innovative approach for microplastic sampling in all surface water bodies using an aquatic drone.

The lack of one standardized method to evaluate microplastic pollution in different aquatic environments worldwide represent a gap to fill for the scientist's community. To help overcome this challenge, we adapted an aquatic drone, named Jellyfishbot®, to sample microplastics. The aquatic drone has been compared with the actual most used method for sampling MPs in surface waters: the Manta net. In order to test the reliability of the aquatic drone in different environments, samples were collected in a river and coastal waters sites. The results obtained with the two methods were similar in term of MPs abundances, shapes and colors. It provides also a better reproducibility and more accurate sampling of MPs located in the surface waters mainly the lighter and smaller ones. This sampling method has the advantage of combining the benefits of Manta net sampling (i.e. a representative surface water sampling method that covers a large sampling area and volume (several tens m3) with those of pump filtration and grab sampling (easy access to confined and hard-to-reach areas). This new sampling method could be applied in different aquatic environments making it possible to compare the data and hence become a new standardized approach to evaluate microplastic pollution levels.

Identification and quantification of plastic additives using pyrolysis-GC/MS: A review.

Analysis of organic plastic additives (OPAs) associated to plastic polymers is growing. The current review outlines the characteristics and the development of (multi-step) pyrolysis coupled with a gas chromatography mass spectrometer (Py-GC/MS) for the identification and semi-quantification of OPAs. Compared to traditional methods, Py-GC/MS offers advantages like suppressing extensive steps of preparation, limiting contamination due to solvents and the possibility to analyse minute particles. Its key advantage is the successive analysis of OPAs and the polymeric matrix of the same sample. Based on the studied articles, numerous methods have been described allowing identification and, in some case, semi-quantification of OPAs. There is nevertheless no gold standard method, especially given the huge diversity of OPAs and the risks of interferences with polymers or other additives, but, among other parameters, a consensus temperature seems to arise from studies. More broadly, this review also explores many aspects on the sample preparation like weight and size of particles and calibration strategies. After studying the various works, some development prospects emerge and it appears that methodological developments should focus on better characterizing the limits of the methods in order to consider which OPAs can be quantified and in which polymers this is feasible.

Occurrence and identification of microplastics in beach sediments from the Hauts-de-France region.

The present work was carried out to quantify microplastics (MP) from three sandy beaches along the Côte d'Opale coastline located in the Hauts-de-France region of northern France. Three different study sites located along the English Channel were investigated due to different levels of anthropopression and hydrodynamic conditions. Sediments were collected at three different tide lines: high tide line (HTL), middle of the intertidal zone (IZ), and low tide line (LTL), to investigate the effects of tide line on microplastic contamination. Particles and fibers were counted and colors were recorded; polymer identification was then performed using pyrolysis-gas chromatography-mass spectrometry (Py-GC/MS). Particle and fiber abundances ranged from 23.4 ± 18.9 to 69.3 ± 30.6 items kg-1 dry weight sediment, with a trend towards fiber predominance, were observed. No difference in particle and fiber abundance was found between the different beaches and tide lines, except for Boulogne-sur-Mer, where the particle number was significantly different between tide lines. Major polymers identified were polyethylene (36.6%) and polypropylene (10.7%). This citizen science project provided preliminary data about the abundance and polymeric nature of MP along the Côte d'Opale coastline.

Development of a qPCR Method for the Identification and Quantification of Two Closely Related Tuna Species, Bigeye Tuna (Thunnus obesus) and Yellowfin Tuna (Thunnus albacares), in Canned Tuna.

Bigeye tuna (Thunnus obesus) and yellowfin tuna (Thunnus albacares) are among the most widely used tuna species for canning purposes. Not only substitution but also mixing of tuna species is prohibited by the European regulation for canned tuna products. However, as juveniles of bigeye and yellowfin tunas are very difficult to distinguish, unintentional substitutions may occur during the canning process. In this study, two mitochondrial markers from NADH dehydrogenase subunit 2 and cytochrome c oxidase subunit II genes were used to identify bigeye tuna and yellowfin tuna, respectively, utilizing TaqMan qPCR methodology. Two different qPCR-based methods were developed to quantify the percentage of flesh of each species used for can processing. The first one was based on absolute quantification using standard curves realized with these two markers; the second one was founded on relative quantification with the universal 12S rRNA gene as the endogenous gene. On the basis of our results, we conclude that our methodology could be applied to authenticate these two closely related tuna species when used in a binary mix in tuna cans.

Identification and mRNA expression of pi-class glutathione S-transferase and selenium-dependent glutathione peroxidase in the gudgeon Gobio gobio exposed to PCB 77.

In aquatic environments some pollutants are present in water and sediments and organisms possess cellular detoxification systems to face up these xenobiotics. The gudgeon, Gobio gobio, is a freshwater benthopelagic fish that appears particularly adequate for an ecotoxicological assessment of rivers. The aim of this study was the identification of GST and GPx genes in this organism in order to develop new indicators of early exposure to xenobiotics in aquatic environments. Reverse-transcription PCR using degenerate primers and RACE-PCR allowed us to identify a selenium-dependent glutathione peroxidase (Se-GPx), belonging to the class one (GPx-1), and a pi-class glutathione S-transferase (pi-GST) cDNAs. These sequences encoded for 191 and 208 amino acids proteins respectively, they exhibit high identities and similarities with corresponding proteins in other fish and the residues essential to the enzymatic functions are highly conserved. The expression patterns of these two mRNA were established by real-time PCR in five gudgeon tissues: liver, gills, kidney, spleen and muscle. The mRNA levels of these genes were highest in the liver and their expression in the other tissues exhibited some differences. Gudgeons exposed to PCB 77 in the food exhibited an increase in pi-GST mRNA and a decrease in GPx-1 transcripts levels in the liver. However, no modification of the enzymatic activities was observed. The present study provides the first transcriptional data regarding pi-GST and GPx-1 in the gudgeon Gobio gobio.

Metallothionein coding sequence identification and seasonal mRNA expression of detoxification genes in the bivalve Corbicula fluminea.

The aim of this study was to identify a metallothionein (MT) coding sequence from the freshwater bivalve Corbicula fluminea and to measure the seasonal transcriptional pattern of MT in parallel with several detoxification genes: superoxide dismutase (SOD), catalase (CAT), glutathione S-transferases (GST) and glutathione peroxidases (GPx), in the digestive gland and the gills of this bivalve during a 1-year period. We identified a C. fluminea MT complete cDNA sequence using RT-PCR and RACE-PCR. The amino acid sequence deduced from the coding sequence encodes for a protein of 73 amino acids containing 21 cysteine residues. This protein exhibits high identities and similarities with the MT sequences of numerous bivalves. MT, SOD, CAT, pi-GST and Se-GPx expression patterns did not exhibit major seasonal variations. A slight increase of MT was observed in July. Therefore, the mRNA expression of these five genes could be used as biomarkers for monitoring studies.

Molecular cloning and expression study of pi-class glutathione S-transferase (pi-GST) and selenium-dependent glutathione peroxidase (Se-GPx) transcripts in the freshwater bivalve Dreissena polymorpha.

Glutathione S-transferases (GST) and glutathione peroxidases (GPx) are essential components of cellular detoxification systems. We identified GST and GPx transcripts in the freshwater bivalve Dreissena polymorpha, their full-length coding sequences were obtained by reverse-transcription PCR using degenerated primers followed by 5' and 3' RACE-PCR (rapid amplification of cDNA ends-PCR). The cDNA identified encoded proteins of 205 and 243 amino acids corresponding respectively to a pi-class GST and a selenium-dependent GPx. The comparison of the deduced amino acid sequences with GST and GPx from other species showed that the residues essential to the enzymatic function of these two proteins are highly conserved. We studied their expression pattern in the digestive gland, the gills and the excretory system of D. polymorpha. The results showed that pi-GST mRNA expression is higher in the digestive gland than in the gills or the excretory system. Se-GPx transcripts are expressed at high, medium and very low levels in the digestive gland, the excretory system and the gills, respectively.

Identification, sequencing and expression of selenium-dependent glutathione peroxidase transcript in the freshwater bivalve Unio tumidus exposed to Aroclor 1254.

Glutathione peroxidases (GPx) and glutathione S-transferases (GST) are essential enzymes of the cellular defense system. The aim of this work was the identification of GPx transcript in a freshwater bivalve, Unio tumidus, and the effects of Aroclor 1254 on GPx and pi-class GST (pi-GST) expression pattern. The GPx full-length coding sequence was obtained by reverse transcription PCR using degenerated primers followed by 5' and 3' rapid amplification of cDNA ends. The GPx cDNA encodes a protein of 232 amino acids. The 72nd amino acid corresponds to a selenocysteine encoded by a TGA codon. Residues essential to the enzymatic function are conserved in GPx of U. tumidus. Specific amplifications of the Se-GPx mRNA from U. tumidus were performed on the digestive gland, the excretory system and the gills. Se-GPx expression level is highest in the digestive gland. No induction of the Se-GPx was observed at the transcriptional level in the digestive gland and the excretory system of Aroclor-treated mussels, while an increase of the pi-GST mRNA level was observed in the excretory system.

cDNA cloning and expression pattern of pi-class glutathione S-transferase in the freshwater bivalves Unio tumidus and Corbicula fluminea.

Glutathione S-transferases (GSTs) are enzymes involved in major detoxification reactions of xenobiotics in many organisms. The aim of this work was the identification of GST transcripts in the freshwater bivalves Unio tumidus and Corbicula fluminea. We used degenerated primers designed in the highly conserved regions of GST to amplify the corresponding mRNA. Full-length coding sequences were obtained by 5' and 3' rapid amplification of cDNA ends. In the two species, the GST cDNAs identified encoded a protein of 205 amino acids. The comparison of the deduced amino acid sequences with GSTs from other species showed that the enzymes belong to the pi-class and the amino acids defining the binding sites of glutathione (G-site) and for xenobiotic substrates (H-site) are highly conserved. Specific amplifications of the GST mRNA from U. tumidus and C. fluminea were performed on the digestive gland, the excretory system and the gills. For each mussel, the results revealed that the pi-class GSTs are expressed at the same level in the three tissues.