Generation of synthetic FTIR spectra to facilitate chemical identification of microplastics.

In a context where learning databases of microplastic FTIR spectra are often incomplete, the objective of our work was to test whether a synthetic data generation method could be relevant to fill the gaps. To this end, synthetic spectra were generated to create new databases. The effectiveness of machine learning from these databases was then tested and compared with previous results. The results showed that the creation of synthetic learning databases could avoid, to a certain extent, the need for learning databases of environmental microplastics FTIR spectra. However, some limitations were encountered, for example, when two different chemical classes had very similar reference spectra or when the intensities of the bands associated with fouling became too intense. The FTIR study of the ageing and fouling of microplastics in the natural environment is one of the identified ways that could further improve this approach.

Bioplastic degradation and assimilation processes by a novel bacterium isolated from the marine plastisphere.

Biosourced and biodegradable plastics offer a promising solution to reduce environmental impacts of plastics for specific applications. Here, we report a novel bacterium named Alteromonas plasticoclasticus MED1 isolated from the marine plastisphere that forms biofilms on foils of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). Experiments of degradation halo, plastic matrix weight loss, bacterial oxygen consumption and heterotrophic biosynthetic activity showed that the bacterial isolate MED1 is able to degrade PHBV and to use it as carbon and energy source. The likely entire metabolic pathway specifically expressed by this bacterium grown on PHBV matrices was shown by further genomic and transcriptomic analysis. In addition to a gene coding for a probable secreted depolymerase, a gene cluster was located that encodes characteristic enzymes involved in the complete depolymerization of PHBV, the transport of oligomers, and in the conversion of the monomers into intermediates of central carbon metabolism. The transcriptomic experiments showed the activation of the glyoxylate shunt during PHBV degradation, setting the isocitrate dehydrogenase activity as regulated branching point of the carbon flow entering the tricarboxylic acid cycle. Our study also shows the potential of exploring the natural plastisphere to discover new bacteria with promising metabolic capabilities.

First national reference of microplastic contamination of French soils.

The recent emergence of studies on plastic contamination of terrestrial environments has revealed the presence of microplastics (MP) in a variety of soil types, from the most densely populated areas to the most remote ones. However, the concentrations and chemical natures of MP in soils vary between studies, and only a few ones have focused on this issue in France. The MICROSOF project aimed to establish the first national references for French soil contamination by microplastics. 33 soil samples randomly chosen on the French soil quality-monitoring network were analyzed. The study collected data on the abundance of microplastics in the [315-5000] µm range, their chemical nature and size, as well as mass abundance estimates and other relevant information. Results demonstrated that 76 % of the soil samples contained microplastics, in concentrations ranging from <6.7 to 80 MP.kg-1 (dry soil). Most samples from croplands, grasslands and vineyards and orchards were contaminated, whereas only one sample from forest contained MP, suggesting an increased risk of microplastic contamination in soils exposed to agricultural practices. The MP abundances are not statistically different from similar studies, indicating an intermediate level of contamination in French soils. Despite intervention reports and surveys, the sources remain unclear at this stage. For the first time, an overview of the state of soil contamination in France, as well as the potential risks is provided.

Sunlight and marine weathering of poly(oxymethylene): Evolution of the physico-chemical properties.

Plastic pollution is now an environmental problem that affects all environmental compartments. The study of plastic degradation in terrestrial, marine and other freshwater environments is emerging. Research is mainly focused on plastic fragmentation into microplastics. In this contribution, an engineering polymer, poly(oxymethylene) (POM), was studied under different weathering conditions using physico-chemical characterization techniques. A POM homopolymer and a POM copolymer were characterized by electron microscopy, tensile tests, DSC, infrared spectroscopy and rheometry tests after climatic and marine weathering or artificial UV/water spray cycles. Natural climatic conditions were the most favorable for POM degradation, especially under solar UV, as evidenced by the strong fragmentation into microplastics when subjected to artificial UV cycles. The evolution of properties with exposure time was found to be non-linear under natural conditions, in contrast to artificial conditions. Two main stages of degradation were evidenced by the correlation between strain at break and carbonyl indices.

Mission Tara Microplastics: a holistic set of protocols and data resources for the field investigation of plastic pollution along the land-sea continuum in Europe.

The Tara Microplastics mission was conducted for 7 months to investigate plastic pollution along nine major rivers in Europe-Thames, Elbe, Rhine, Seine, Loire, Garonne, Ebro, Rhone, and Tiber. An extensive suite of sampling protocols was applied at four to five sites on each river along a salinity gradient from the sea and the outer estuary to downstream and upstream of the first heavily populated city. Biophysicochemical parameters including salinity, temperature, irradiance, particulate matter, large and small microplastics (MPs) concentration and composition, prokaryote and microeukaryote richness, and diversity on MPs and in the surrounding waters were routinely measured onboard the French research vessel Tara or from a semi-rigid boat in shallow waters. In addition, macroplastic and microplastic concentrations and composition were determined on river banks and beaches. Finally, cages containing either pristine pieces of plastics in the form of films or granules, and others containing mussels were immersed at each sampling site, 1 month prior to sampling in order to study the metabolic activity of the plastisphere by meta-OMICS and to run toxicity tests and pollutants analyses. Here, we fully described the holistic set of protocols designed for the Mission Tara Microplastics and promoted standard procedures to achieve its ambitious goals: (1) compare traits of plastic pollution among European rivers, (2) provide a baseline of the state of plastic pollution in the Anthropocene, (3) predict their evolution in the frame of the current European initiatives, (4) shed light on the toxicological effects of plastic on aquatic life, (5) model the transport of microplastics from land towards the sea, and (6) investigate the potential impact of pathogen or invasive species rafting on drifting plastics from the land to the sea through riverine systems.

Continents of plastics: An estimate of the stock of microplastics in agricultural soils.

While there are estimates of the stock of microplastics in the marine environment, there are no estimates for soils. The main objective of this work is to estimate the total mass of microplastics in global agricultural soils. Microplastic abundance data from 442 sampling sites were collected from 43 articles. From these, the median of the abundance values, as well as the abundance profile of microplastics in soils were calculated. Thus, 1.5 to 6.6 Mt of microplastics would be present in soils on a global scale, i.e. one to two orders of magnitude higher than the estimated ocean surface microplastic stock. However, many limitations exist to accurately calculate these stocks. This work should therefore be considered as a first step in addressing this question. In the long term, in order to better assess this stock, it seems important to obtain more diversified data, e.g. better representing certain countries, or certain land uses.

A bionanocomposite of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)/ZnO-nanoparticles intended for food packaging.

Films-based bionanocomposites have gained a great importance in food plastic packaging because they are eco-friendly materials and have the potential to improve food protection, while limiting the accumulation of synthetic plastics on the planet. In this paper, biofilms were prepared using poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) reinforced with Zinc oxide nanoparticles (ZnO-NPs) to develop new bionanocomposite materials intended for food packaging. The samples were fabricated using first solvent casting method followed by melt compounding at various loading rates, i.e., 1.5, 3 and 6 wt%. The obtained results showed that the incorporation of ZnO-NPs to PHBHHx at 3 wt% leads to higher crystallinity, improved mechanical properties and antimicrobial activity, compared with neat polymer and other bionanocomposites. This was attributed to the finer and homogeneous nanofiller dispersion in the polymer matrix evidenced by scanning electron microscopy analysis. Whereas at 6 wt%, the bionanocomposite sample exhibited low mechanical properties due to the formation of ZnO-NPs aggregates. In view of the obtained results, the study highlights the potential of using the PHBHHx/ZnO-NPs bionanocomposite at 3 wt% in food packaging without any prior filler treatment.

Discussion about suitable applications for biodegradable plastics regarding their sources, uses and end of life.

This opinion paper offers a scientific view on the current debate of the place of biodegradable plastics as part of the solution to deal with the growing plastic pollution in the world's soil, aquatic, and marine compartments. Based on the current scientific literature, we focus on the current limits to prove plastic biodegradability and to assess the toxicity of commercially used biobased and biodegradable plastics in natural environments. We also discuss the relevance of biodegradable plastics for selected applications with respect to their use and end of life. In particular, we underlined that there is no universal biodegradability of plastics in any ecosystem, that considering the environment as a waste treatment system is not acceptable, and that the use of compostable plastics requires adaptation of existing organic waste collection and treatment channels.

Microplastic fouling: A gap in knowledge and a research imperative to improve their study by infrared characterization spectroscopy.

The marine weathering of microplastics is spectrally characterized by the appearance of new bands that disturb our understanding of the information carried by the spectra. Yet, no explanation has been provided on the chemical origin of these new bands. Thus, the main objective of this work was to identify the origins of these additional bands. To this end, 4042 spectra of poly (styrene), poly(ethylene) and poly(propylene) microplastics collected in the Mediterranean Sea, were analysed using principal component analysis. The results showed that the spectral variability was mainly related to only three processes: chemical ageing, organic and inorganic fouling. These processes probably differ from one polymer family to another due to surface affinities. This work has also led to the proposal of two new polymer indices that could be used to monitor the intensity of (bio)fouling. Finally, the development of advanced analyses could also provide information on the nature of the plastisphere.

Seasonal patterns of microplastics in surface sediments of a Mediterranean lagoon heavily impacted by human activities (Bizerte lagoon, Northern Tunisia).

In this paper, we investigated seasonal variations in concentrations of microplastics (MPs) in surface sediments of a lagoon heavily impacted by human activities, located in northern Tunisia (the Bizerte lagoon, southern Mediterranean Sea). Analyses of 112 sediment samples collected from 28 stations between May 2019 and February 2020 revealed significant seasonal variation in concentrations of total MPs, with the highest levels recorded in August 2019 (109.6 ± 59.8 items kg-1 DS (dry sediment)) and the lowest in February 2020 (33.2 ± 22.0 items kg-1 DS). In terms of polymer types, polyethylene particles were the most abundant throughout the year, followed by polypropylene, polyvinyl chloride, and polyethylene terephthalate. Spatial variations in total MP concentrations were found to depend on several environmental factors, including proximity to the coastline, level of anthropogenic pressure, location inside the lagoon, and presence/absence of vegetation. The upper 5-cm surface sediment layer of the lagoon was found to contain ~ 9.96 × 1010 MP particles, equal to ~ 248.97 t of plastic. Similar patterns of microplastic composition and structure were found throughout the year, revealing the same plastic pollution hotspots during all seasons. This indicates that sources of plastic pollution are land-based and originate from coastal urban, industrial, and agricultural areas, as well as from major freshwater streams. The findings of the present work can help to develop an efficient environmental management plan aiming to reduce and/or stop the spread of plastic pollution and its impacts on the socially and economically important ecosystem of the Bizerte lagoon.

Chemical composition of microplastics floating on the surface of the Mediterranean Sea.

The Mediterranean Sea is one of the most studied regions in the world in terms of microplastic (MP) contamination. However, only a few studies have analysed the chemical composition of MPs at the Mediterranean Sea surface. In this context, this study aims to describe the chemical composition as a function of particle size, mass and number concentrations of MPs collected in the surface waters of the Mediterranean Sea. The chemical composition showed a certain homogeneity at the Mediterranean Sea scale. The main polymers identified by Fourier Transform Infra-Red (FTIR) spectroscopy were poly(ethylene) (67.3 ± 2.4%), poly(propylene) (20.8 ± 2.1%) and poly(styrene) (3.0 ± 0.9%). Nevertheless, discrepancies, confirmed by the literature, were observed at a mesoscale level. Thus, in the North Tyrrhenian Sea, the proportion of poly(ethylene) was significantly lower than the average value of the Mediterranean Sea (57.9 ± 10.5%). Anthropic sources, rivers, or polymer ageing are assumed to be responsible for the variations observed.

Bacterial Abundance, Diversity and Activity During Long-Term Colonization of Non-biodegradable and Biodegradable Plastics in Seawater.

The microorganisms living on plastics called "plastisphere" have been classically described as very abundant, highly diverse, and very specific when compared to the surrounding environments, but their potential ability to biodegrade various plastic types in natural conditions have been poorly investigated. Here, we follow the successive phases of biofilm development and maturation after long-term immersion in seawater (7 months) on conventional [fossil-based polyethylene (PE) and polystyrene (PS)] and biodegradable plastics [biobased polylactic acid (PLA) and polyhydroxybutyrate-co-hydroxyvalerate (PHBV), or fossil-based polycaprolactone (PCL)], as well as on artificially aged or non-aged PE without or with prooxidant additives [oxobiodegradable (OXO)]. First, we confirmed that the classical primo-colonization and growth phases of the biofilms that occurred during the first 10 days of immersion in seawater were more or less independent of the plastic type. After only 1 month, we found congruent signs of biodegradation for some bio-based and also fossil-based materials. A continuous growth of the biofilm during the 7 months of observation (measured by epifluorescence microscopy and flow cytometry) was found on PHBV, PCL, and artificially aged OXO, together with a continuous increase in intracellular (3H-leucine incorporation) and extracellular activities (lipase, aminopeptidase, and ß-glucosidase) as well as subsequent changes in biofilm diversity that became specific to each polymer type (16S rRNA metabarcoding). No sign of biodegradation was visible for PE, PS, and PLA under our experimental conditions. We also provide a list of operational taxonomic units (OTUs) potentially involved in the biodegradation of these polymers under natural seawater conditions, such as Pseudohongiella sp. and Marinobacter sp. on PCL, Marinicella litoralis and Celeribacter sp. on PHBV, or Myxococcales on artificially aged OXO. This study opens new routes for a deeper understanding of the polymers' biodegradability in seawaters, especially when considering an alternative to conventional fossil-based plastics.

Microbial Diversity and Activity During the Biodegradation in Seawater of Various Substitutes to Conventional Plastic Cotton Swab Sticks.

The European Parliament recently approved a new law banning single-use plastic items for 2021 such as plastic plates, cutlery, straws, cotton swabs, and balloon sticks. Transition to a bioeconomy involves the substitution of these banned products with biodegradable materials. Several materials such as polylactic acid (PLA), polybutylene adipate terephthalate (PBAT), poly(butylene succinate) (PBS), polyhydroxybutyrate-valerate (PHBV), Bioplast, and Mater-Bi could be good candidates to substitute cotton swabs, but their biodegradability needs to be tested under marine conditions. In this study, we described the microbial life growing on these materials, and we evaluated their biodegradability in seawater, compared with controls made of non-biodegradable polypropylene (PP) or biodegradable cellulose. During the first 40 days in seawater, we detected clear changes in bacterial diversity (Illumina sequencing of 16S rRNA gene) and heterotrophic activity (incorporation of 3H-leucine) that coincided with the classic succession of initial colonization, growth, and maturation phases of a biofilm. Biodegradability of the cotton swab sticks was then tested during another 94 days under strict diet conditions with the different plastics as sole carbon source. The drastic decrease of the bacterial activity on PP, PLA, and PBS suggested no bacterial attack of these materials, whereas the bacterial activity in PBAT, Bioplast, Mater-Bi, and PHBV presented similar responses to the cellulose positive control. Interestingly, the different bacterial diversity trends observed for biodegradable vs. non-biodegradable plastics allowed to describe potential new candidates involved in the degradation of these materials under marine conditions. This better understanding of the bacterial diversity and activity dynamics during the colonization and biodegradation processes contributes to an expanding baseline to understand plastic biodegradation in marine conditions and provide a foundation for further decisions on the replacement of the banned single-used plastics.

Pre-detection of microplastics using active thermography.

Spectrometric analysis is one of the most widely used approaches to characterize the chemical nature of microplastics. Despite recent developments, this key step remains time consuming. The aim of this paper is to propose a new method for the pre-detection of microplastics based on mid-infrared imaging. Plastic particles were mixed with sand particles and placed on a glass filter. Infrared observation with a thermal camera shows a stronger thermal contrast measured between the filter and the plastics than between the filter and the sand, which reveals the plastic particles in a few tens of seconds. An image processing tool is then used to amplify this contrast. Furthermore, this pre-detection method makes it possible to propose hypotheses on the most probable chemical nature of the particles identified. Consequently, pre-detection using active thermography constitutes a promising way of significantly accelerating microplastic study.

Characterization of microplastics in the surface waters of an urban lagoon (Bizerte lagoon, Southern Mediterranean Sea): Composition, density, distribution, and influence of environmental factors.

Compared to open sea environments, there is still limited knowledge about microplastic levels in semi-enclosed systems such as coastal lagoons. This work aims to assess the levels of MPs in the waters of an urban lagoon (Bizerte lagoon, northern Tunisia) and to study the effects of environmental factors on their distribution and abundance. Average concentration of total MPs was found to be 453.0 ± 335.2 items m-3. The upper 25 cm water layer of the lagoon is most likely to contain ~16.99 × 109 MPs items (which correspond to a total mass of 42.47 t). Fibers were the primary MPs types encountered, and most of MPs particles identified were white and clear-colored. Polyethylene and polypropylene were the predominant polymer types in Bizerte lagoon. Among the various potential plastic sources of MPs, unmanaged domestic plastic wastes are likely to be the major source of plastic pollution in the lagoon. Several environmental factors appeared to influence the distribution and density of MPs in the lagoon waters. These information contribute to better understand the dynamics of MPs in lagoons and to develop environmental management actions.

Microplastics in edible mussels from a southern Mediterranean lagoon: Preliminary results on seawater-mussel transfer and implications for environmental protection and seafood safety.

This study assesses the microplastics (MPs) levels in mussels (Mytilus galloprovincialis) and seawater from a southern Mediterranean lagoon (Bizerte lagoon, Northern Tunisia) and discusses the potential effects of its consumption on human health. Polyethylene was the most abundant in mussels and seawater, followed by polypropylene and cellophane. The lowest and highest average MPs concentrations were recorded in the lagoon channel and southern area of the lagoon, respectively, for both mussels (2.6 ± 1.7-12.0 ± 1.4 items mussel-1) and seawater (0.2 ± 0.1-0.7 ± 0.2 items L-1). Mussels in areas highly polluted with fibers and polyethylene were found to have higher potential to ingest and/or adhere higher numbers of these particles. The annual dietary intake of MPs by Tunisians through the consumption of local mussels was estimated at 4.2 items capita-1 year-1. Even though MPs are not biodegraded and can be excreted by humans, their potential human health risks are discussed in this paper.

Why is there plastic packaging in the natural environment? Understanding the roots of our individual plastic waste management behaviours.

Plastic waste is now a classic contaminant of the natural environment and the origins of the contamination need to be well understood. The transition from a useful object to a waste product is a fundamental moment that, from the point of view of the scientific literature, remains poorly understood. This review therefore aims to highlight some factors controlling this intentionality, but also those that influence individual waste management behaviours. For this purpose, an original approach involving the study of the amount of knowledge within different disciplinary fields of research has been employed. The results underline that the low direct impact of the consequences on their users of the discarding of plastic packaging seems to be an important reason for individual mismanagement. Furthermore, the modern individual behaviours of the discarding of plastics are often deeply rooted in the past of the populations. Policies to reduce waste disposal come up against strong individual behavioural constraints that limit the proper management of plastic waste. Thus, incivilities, difficulty in enforcing sanctions, or public opposition to changes in waste management are all factors that contribute to the maintenance waste discarding behaviour. The reuse behaviour of objects that have become useless is also historically attested, but has tended to disappear with the rise of the consumer society. This type of behaviour, whose valorisation is a way of reducing plastic waste abandonment behaviour, remains, however, less scientifically studied than other ways such as recycling.

PHA Production and PHA Synthases of the Halophilic Bacterium Halomonas sp. SF2003.

Among the different tools which can be studied and managed to tailor-make polyhydroxyalkanoates (PHAs) and enhance their production, bacterial strain and carbon substrates are essential. The assimilation of carbon sources is dependent on bacterial strain's metabolism and consequently cannot be dissociated. Both must wisely be studied and well selected to ensure the highest production yield of PHAs. Halomonas sp. SF2003 is a marine bacterium already identified as a PHA-producing strain and especially of poly-3-hydroxybutyrate (P-3HB) and poly-3-hydroxybutyrate-co-3-hydroxyvalerate (P-3HB-co-3HV). Previous studies have identified different genes potentially involved in PHA production by Halomonas sp. SF2003, including two phaC genes with atypical characteristics, phaC1 and phaC2. At the same time, an interesting adaptability of the strain in front of various growth conditions was highlighted, making it a good candidate for biotechnological applications. To continue the characterization of Halomonas sp. SF2003, the screening of carbon substrates exploitable for PHA production was performed as well as production tests. Additionally, the functionality of both PHA synthases PhaC1 and PhaC2 was investigated, with an in silico study and the production of transformant strains, in order to confirm and to understand the role of each one on PHA production. The results of this study confirm the adaptability of the strain and its ability to exploit various carbon substrates, in pure or mixed form, for PHA production. Individual expression of PhaC1 and PhaC2 synthases in a non-PHA-producing strain, Cupriavidus necator H16 PHB¯4 (DSM 541), allows obtaining PHA production, demonstrating at the same time, functionality and differences between both PHA synthases. All the results of this study confirm the biotechnological interest in Halomonas sp. SF2003.

Relative Influence of Plastic Debris Size and Shape, Chemical Composition and Phytoplankton-Bacteria Interactions in Driving Seawater Plastisphere Abundance, Diversity and Activity.

The thin film of life that inhabits all plastics in the oceans, so-called "plastisphere," has multiple effects on the fate and impacts of plastic in the marine environment. Here, we aimed to evaluate the relative influence of the plastic size, shape, chemical composition, and environmental changes such as a phytoplankton bloom in shaping the plastisphere abundance, diversity and activity. Polyethylene (PE) and polylactide acid (PLA) together with glass controls in the forms of meso-debris (18 mm diameter) and large-microplastics (LMP; 3 mm diameter), as well as small-microplastics (SMP) of 100 µm diameter with spherical or irregular shapes were immerged in seawater during 2 months. Results of bacterial abundance (confocal microscopy) and diversity (16S rRNA Illumina sequencing) indicated that the three classical biofilm colonization phases (primo-colonization after 3 days; growing phase after 10 days; maturation phase after 30 days) were not influenced by the size and the shape of the materials, even when a diatom bloom (Pseudo-nitzschia sp.) occurred after the first month of incubation. However, plastic size and shape had an effect on bacterial activity (3H leucine incorporation). Bacterial communities associated with the material of 100 µm size fraction showed the highest activity compared to all other material sizes. A mature biofilm developed within 30 days on all material types, with higher bacterial abundance on the plastics compared to glass, and distinct bacterial assemblages were detected on each material type. The diatom bloom event had a great impact on the plastisphere of all materials, resulting in a drastic change in diversity and activity. Our results showed that the plastic size and shape had relatively low influence on the plastisphere abundance, diversity, and activity, as compared to the plastic composition or the presence of a phytoplankton bloom.

A machine learning algorithm for high throughput identification of FTIR spectra: Application on microplastics collected in the Mediterranean Sea.

The development of methods to automatically determine the chemical nature of microplastics by FTIR-ATR spectra is an important challenge. A machine learning method, named k-nearest neighbors classification, has been applied on spectra of microplastics collected during Tara Expedition in the Mediterranean Sea (2014). To realize these tests, a learning database composed of 969 microplastic spectra has been created. Results show that the machine learning process is very efficient to identify spectra of classical polymers such as poly(ethylene), but also that the learning database must be enhanced with less common microplastic spectra. Finally, this method has been applied on more than 4000 spectra of unidentified microplastics. The verification protocol showed less than 10% difference in the results between the proposed automated method and a human expertise, 75% of which can be very easily corrected.