Identification and Quantification of Microplastics in the Marine Environment Using the Laser Direct Infrared (LDIR) Technique.

Here, we evaluate for the first time the performances of the newly developed laser direct infrared (LDIR) technique and propose an optimization of the initial protocol for marine microplastics (MPs) analysis. Our results show that an 8 µm porosity polycarbonate filter placed on a Kevley slide enables preconcentration and efficient quantification of MPs, as well as polymer and size determination of reference plastic pellets of polypropylene (PP), polyethylene (PE), polystyrene (PS), polyvinyl chloride (PVC), and polyethylene terephthalate (PET), with recoveries ranging from 80-100% and negligible blank values for particle sizes ranging from 200 to 500 µm. A spiked experiment using seawater, sediment, mussels, and fish stomach samples showed that the method responded linearly with significant slopes (R2 ranging from 0.93-1.0; p < 0.001, p < 0.01). Overall, 11 polymer types were identified with limited handling and an analysis time of ca. 3 h for most samples and 6 h for complex samples. Application of this technique to Mediterranean marine samples (seawater, sediment, fish stomachs and mussels) indicated MP concentrations and size distribution consistent with the literature. A high predominance of PVC (sediment, fish stomachs) and PE and PP (seawater, mussels) was observed in the analyzed samples.

Microplastic abundance and characteristics in French Atlantic coastal sediments using a new extraction method.

The ubiquitous presence of microplastics (MPs) has been demonstrated in all environmental compartments in the recent years. They are detected in air, freshwater, soil, organisms and particularly in marine ecosystems. Since sediments are known to be the major sink of many organic and inorganic pollutants, the aim of this study was to develop and validate a fast and cheap methodology to assess the MP contamination in intertidal sediments from the Gulf of Biscay (Pays de la Loire region, France). Sediments were sampled at three locations (Pays de la Loire region, France) and during two seasons: October 2015 and March 2016. The analytical protocol involved MP extraction from dried sediments using milliQ water and a centrifugation technique. After a filtration step of supernatants, MPs were detected and directly identified on the membrane filters using µFTIR spectroscopy in reflection mode. For the first time, the number of replicates allowing to obtain a satisfying representativeness of the whole sampled sediment was also evaluated at 10 replicates of 25 g each. The average number of MPs in sediments was 67 (±76) MPs/kg dw (N = 60) with no significant difference between sites and seasons. Ten different compositions of MPs were defined by µFT-IR with a high proportion of polypropylene (PP) and polyethylene (PE), 38 and 24%, respectively. Among MPs, mainly fragments (84%) were observed with main size classes corresponding to [>100 µm] and [50-100 µm] but no particles > 1 mm could be found suggesting that mainly small microplastics (<1 mm) were subject to vertical transport.

Quantification and characterization of microplastics in blue mussels (Mytilus edulis): protocol setup and preliminary data on the contamination of the French Atlantic coast.

Microplastics (MPs) constitute a main environmental issue due to their threat to marine organisms and so far to humans. The lack of a fast standard protocol in MP isolation and identification from living organisms bring to challenge for the science. In this paper, an optimized protocol using potassium hydroxide 10% (KOH 10%; m/v) for digestion of mussel soft tissues (Mytilus edulis) and multi-steps of sedimentation has been developed. Efficiency higher than 99.9% of organic and mineral matter elimination was shown by application on mussels sampled on the French Atlantic coast. The identification of MPs was performed by FTIR microscopy straight on the filter and the whole analysis can be compatible with a routine goal. Fourteen MPs of four different chemical natures were found and identified in 5 pools of 3 sampled mussels. Their size ranged from 30 to 200 µm. Further investigations are now needed to evaluate the potential risk of such particles within this marine bivalve species and other filter feeders.

Factors influencing the microplastic contamination of bivalves from the French Atlantic coast: Location, season and/or mode of life?

Monitoring the presence of microplastics (MP) in marine organisms is currently of high importance. This paper presents the qualitative and quantitative MP contamination of two bivalves from the French Atlantic coasts: the blue mussel (Mytilus edulis) and the Pacific oyster (Crassostrea gigas). Three factors potentially influencing the contamination were investigated by collecting at different sampling sites and different seasons, organisms both wild and cultivated. Inter- and intra-species comparisons were also achieved. MP quantity in organisms was evaluated at 0.61±0.56 and 2.1±1.7MP per individual respectively for mussels and oysters. Eight different polymers were identified. Most of the MPs were fragments; about a half of MPs were grey colored and a half with a size ranging from 50 to 100µm for both studied species. Some inter-specific differences were found but no evidence for sampling site, season or mode of life effect was highlighted.

Is there any consistency between the microplastics found in the field and those used in laboratory experiments?

The ubiquitous presence and persistency of microplastics (MPs) in aquatic environments are of particular concern since they represent an increasing threat to marine organisms and ecosystems. Great differences of concentrations and/or quantities in field samples have been observed depending on geographical location around the world. The main types reported have been polyethylene, polypropylene, and polystyrene. The presence of MPs in marine wildlife has been shown in many studies focusing on ingestion and accumulation in different tissues, whereas studies of the biological effects of MPs in the field are scarce. If the nature and abundance/concentrations of MPs have not been systematically determined in field samples, this is due to the fact that the identification of MPs from environmental samples requires mastery and execution of several steps and techniques. For this reason and due to differences in sampling techniques and sample preparation, it remains difficult to compare the published studies. Most laboratory experiments have been performed with MP concentrations of a higher order of magnitude than those found in the field. Consequently, the ingestion and associated effects observed in exposed organisms have corresponded to great contaminant stress, which does not mimic the natural environment. Medium contaminations are produced with only one type of polymer of a precise sizes and homogenous shape whereas the MPs present in the field are known to be a mix of many types, sizes and shapes of plastic. Moreover, MPs originating in marine environments can be colonized by organisms and constitute the sorption support for many organic compounds present in environment that are not easily reproducible in laboratory. Determination of the mechanical and chemical effects of MPs on organisms is still a challenging area of research. Among the potential chemical effects it is necessary to differentiate those related to polymer properties from those due to the sorption/desorption of organic compounds.

Development and validation of LC-MS methods for peptaibol quantification in fungal extracts according to their lengths.

Some terrestrial Trichoderma sp. strains are already used as biological control agents (BCAs). They all produce peptaibols, small antimicrobial peptides which are supposed to play a role in the anti-phytopathogenic activity of Trichoderma sp. Trichoderma strains producing high amounts of peptaibols could represent new potential BCAs. In this context, marine-derived Trichoderma strains from the marine fungal strain collection of the "Mer, Molécules, Santé" (MMS) laboratory were investigated for their peptaibol production. Previously, the quantification of peptaibols was performed using alamethicin, as standard (20-amino acid residues peptaibol). In this study, the development and validation of quantification LC/ESI-TI-MS methods using different standards of peptaibols (11-, 14- and 20-amino acid residues) was performed in order to quantify all of them, in a single analysis, in Trichoderma crude extracts according to their chain length. The developed and validated methods were used to study the peptaibol production kinetic of a marine-derived Trichoderma strain, i.e., Trichoderma longibrachiatum (MMS 151). The results showed the optimal culture time at the 9th day with concentrations reaching 1.4±0.2% and 2.3±0.4% of the fungal biomass respectively for 11- and 20-residue peptaibols. Then, the different peptaibol subgroups produced by 13 Trichoderma strains were quantified. According to their 18-, 19- and 20-residue peptaibol production, three strains referenced as MMS 1541, MMS 639 and MMS 151 seemed to be good candidates as potential new biological control agents with respective production of 0.4, 0.4 and 2.1%.