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Every year, rivers introduce a staggering amount of hundred kilotons of plastic into the Oceans. This plastic is inhabited by microorganisms known as the plastisphere, which can be transferred between different ecosystems through the transport of microplastics. Here, we simulated the microbial colonization of polyethylene-based plastic pellets that are classically used to manufacture large-scale plastic products. The pellets were immersed for 1 month in four to five sampling stations along the river-to-sea continuum of nine of the major European rivers. This study presents the first untargeted metabolomics analysis of the plastisphere, by using ultra high-performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS). The plastisphere metabolomes were similar in the Rhine and Rhone rivers, while being different from the Tiber and Loire rivers, which showed greater similarity to the Thames and Seine rivers. Interestingly, we found a clear distinction between plastisphere metabolomes from freshwater and marine water in most of the river-to-sea continuum, thus suggesting a complete segregation in plastisphere metabolites that is not consistent with a major transfer of microorganisms between the two contrasted ecosystems. Putative annotations of 189 discriminating metabolites suggested that lipid metabolism was significantly modulated. These results enlightened the relevance of using environmental metabolomic as complementary analysis to the current OMICs analysis.
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Styrene butadiene rubber is one of the main constituents of tire tread. During tire life, the tread material undergoes different stresses that impact its structure and chemical composition. Wear particles are then released into the environment as weathered material. To understand their fate, it is important to start with a better characterization of abiotic and biotic degradation of the elastomer material. A multi-disciplinary approach was implemented to study the photo- and thermo- degradation of non-vulcanized SBR films containing 15 w% styrene as well as their potential biodegradation by Rhodoccocus ruber and Gordonia polyisoprenivorans bacterial strains. Each ageing process leads to crosslinking reactions, much surface oxidation of the films and the production of hundreds of short chain compounds. These degradation products present a high level of unsaturation and oxidation and can be released into water to become potential substrates for microorganisms. Both strains were able to degrade from 0.2 to 1.2 % (% ThOD) of the aged SBR film after 30-day incubation while no biodegradation was observed on the pristine material. A 25-75 % decrease in the signal intensity of water extractable compounds was observed, suggesting that biomass production was linked to the consumption of low-molecular-weight degradation products. These results evidence the positive impact of abiotic degradation on the biodegradation process of styrene butadiene rubber.
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Butadienos , Elastômeros , Borracha , Estirenos , Estireno , ÁguaRESUMO
Roadway particles (RP) that can be collected with on-vehicle system, consist of a mixture of Tire and road wear particles (TRWP) with other traffic-derived particles (exhaust or non-exhaust) and/or biogenic compounds and represent a significant source of xenobiotics, susceptible to reach the different environmental compartments. The study of the RP fate is thus a major challenge to tackle in order to understand their degradation and impact. They offer a variety of carbon sources potentially usable by microorganisms, ranging from the tire-derived plasticizers, vulcanizing agents, protective agents and their transformation products, to other traffic, road and environmental-derived contaminants. A multi-analytical approach was implemented to characterize RP and study their biodegradation. Kinetics of RP extractions were monitored during 21 days in water, methanol, acetone and chloroform to identify leaching and extractable compounds and monitor the particle composition. The results confirmed that hundreds of readily leachable chemicals can be extracted from RP directly into water according to a dynamic process with time while additional poorly soluble compounds remain in the particles. Mass spectrometry (LC-HRMS and GC-MS) allowed us to propose 296 putative compounds using an extensive rubber database. The capacity of 6 bacterial strains, belonging to Rhodococcus, Pseudomonas and Streptomyces genera, to biodegrade RP was then evaluated over 14 days of incubation. The selected strains were able to grow on RP using various substrates. Elastomer monitoring by 1H NMR revealed a significant 12 % decrease of the extractable SBR fraction when the particles were incubated with Rhodococcus ruber. After incubation, the biodegradation of 171 compounds among leachable and extractable compounds was evaluated. Fatty acids and alkanes from rubber plasticizers and paraffin waxes were the most degraded putative compounds by the six strains tested, reaching 75 % of biodegradation for some of them.
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Plastificantes , Borracha , Plastificantes/análise , Emissões de Veículos/análise , Cromatografia Gasosa-Espectrometria de Massas , ÁguaRESUMO
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.
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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.
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Plastics are ubiquitous in the oceans and constitute suitable matrices for bacterial attachment and growth. Understanding biofouling mechanisms is a key issue to assessing the ecological impacts and fate of plastics in marine environment. In this study, we investigated the different steps of plastic colonization of polyolefin-based plastics, on the first one hand, including conventional low-density polyethylene (PE), additivated PE with pro-oxidant (OXO), and artificially aged OXO (AA-OXO); and of a polyester, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), on the other hand. We combined measurements of physical surface properties of polymers (hydrophobicity and roughness) with microbiological characterization of the biofilm (cell counts, taxonomic composition, and heterotrophic activity) using a wide range of techniques, with some of them used for the first time on plastics. Our experimental setup using aquariums with natural circulating seawater during 6 weeks allowed us to characterize the successive phases of primo-colonization, growing, and maturation of the biofilms. We highlighted different trends between polymer types with distinct surface properties and composition, the biodegradable AA-OXO and PHBV presenting higher colonization by active and specific bacteria compared to non-biodegradable polymers (PE and OXO). Succession of bacterial population occurred during the three colonization phases, with hydrocarbonoclastic bacteria being highly abundant on all plastic types. This study brings original data that provide new insights on the colonization of non-biodegradable and biodegradable polymers by marine microorganisms.
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Plant protection based on novel alternative strategies is a major concern in agriculture to sustain pest management. The marc extract of red grape cultivars reveals plant defence inducer properties. Treatment with grape marc extract efficiently induced hypersensitive reaction-like lesions with cell death evidenced by Evans Blue staining of tobacco leaves. Examination of the infiltration zone and the surrounding areas under UV light revealed the accumulation of autofluorescent compounds. Both leaf infiltration and a foliar spray of the red grape extract on tobacco leaves induced defence gene expression. The PR1 and PR2 target genes were upregulated locally and systemically in tobacco plants following grape marc extract treatment. The grape extract elicited an array of plant defence responses making this natural compound a potential phytosanitary product with a challenging issue and a rather attractive option for sustainable agriculture and environmentally friendly practices.
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Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Nicotiana/genética , Extratos Vegetais/toxicidade , Folhas de Planta/química , Vitis/química , Morte Celular/efeitos dos fármacos , Genes de Plantas , RNA de Plantas/genética , Transdução de Sinais , Nicotiana/efeitos dos fármacos , Nicotiana/metabolismo , Regulação para CimaRESUMO
Pesticide reactivity toward light is rarely considered at the leaf surface after crop treatment; regardless, these degradation reactions directly impact the pesticide effectiveness. The use of sunscreen adjuvants to overcome photodegradation has presented some limitations so far. Raw hydroalcoholic plant extracts have been recently proposed to be used as photoprotecting adjuvants; on a model system they significantly decreased the photodegradation of pesticide. Here it is demonstrated that their use makes possible a dose reduction. Sulcotrione, a selective herbicide for use in maize, was tested in a growth chamber equipped with simulated solar light against a typical weed in maize. Sprayed weeds were monitored by biometrical and physiological parameters. Sulcotrione minimum dose required for a good herbicidal efficacy (ED(50), corresponding to 50% of chlorophyll content decay) was estimated to be 55 g ha(-1). In the presence of grape extract added in a 3-fold excess compared to the herbicide, the ED(50) decreased to 34 g ha(-1). The use of grape extract allows extension of sulcotrione herbicidal activity and reduction of the dose by 35% in controlled conditions. This is a promising result for the effective dose field adjustment.
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Adjuvantes Farmacêuticos/química , Química Farmacêutica/métodos , Cicloexanonas/química , Herbicidas/química , Mesilatos/química , Fotólise , Extratos Vegetais/química , Cinética , Vitis/químicaRESUMO
A pesticide's reactivity toward light at the leaf surface after crop treatment is rarely considered, although such degradation reactions directly affect the pesticide's effectiveness. To overcome these limitations, the use of plant pigments was proposed as a new class of photoprotecting agent. The photoprotecting properties of seven plant pigments were tested under controlled conditions over herbicide sulcotrione. Grape wine extracts were tested over a panel of pesticides from distinct chemical families. The addition of plant extracts almost systematically reduced the pesticide's photoreactivity. The grape wine extracts improve at least by 38% the half-life of photolysis of almost all of the active ingredients tested, except for the herbicide triclopyr. Fustictree extract increases by 82% the photostability of the herbicide sulcotrione. Plant extracts mainly act as sunscreens; that is, the photostabilization of the active ingredient is due to the competitive energy absorption of UV photon. The use of natural plant extracts is a promising strategy to limit pesticide photodegradation. It is a way to develop sustainable and innovative technology for the plant protection industry, being beneficial from both economic and ecological points of view.
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Praguicidas/química , Extratos Vegetais/farmacologia , Ecologia , FotoquímicaRESUMO
Photolyses of three herbicides, bentazon, clopyralid, and triclopyr, were studied on plant leaves after crop treatment. The experiments were carried out on cuticular wax films, which are good models for leaf surfaces. The pure compounds and their commercial formulations were investigated under simulated solar light. At the recommended agricultural application rates, the three formulated herbicides photolyzed more rapidly on films than on soil or in water. Their photolysis is likely to be an important dissipation path from crops after treatment. The effects induced by the adjuvants in formulations were varied. Adjuvants slowed the photodegradation of bentazon slightly. In Garlon, in which triclopyr and clopyralid are combined, the adjuvants did not affect the photolysis of clopyralid even though they accelerated the rate of photolysis of triclopyr by a factor of 7. The kinetics were also affected by the application rates. This work also underscores the importance of assessing the photoreactivity of active ingredients in conditions similar to those of their application.