UV Dosage Unveils Toxic Properties of Weathered Commercial Bioplastic Bags.

Previous studies indicated that weathered conventional plastics and bioplastics pose ecotoxicological risks. Here, the effects of artificial and natural weathering on the ecotoxicity of three compostable bags and a conventional polyethylene (PE) bag are investigated. With that aim, a 21-day artificial indoor weathering experiment featuring UV light, UV-filtered light, and darkness was run simultaneously to a 120-day outdoor littoral mesocosm exposure featuring natural light, UV-filtered light, and shaded conditions. Acute toxicity of so-weathered plastic specimens was tested in vivo using the sensitive Paracentrotus lividus sea-urchin embryo test. PE was nontoxic from the beginning and did not gain toxicity due to UV weathering. In contrast, for bioplastics, dry artificial UV weathering increased toxicity in comparison to the dark control. Weathering in outdoor mesocosm led to a rapid loss of toxic properties due to leaching in rainwater. With a higher UV dosage, a plastic-type-dependent regain of toxicity was observed, most likely driven by enhanced availability or transformation of functional additives or due to bioplastic degradation products. PE showed moderate UV absorbance, while bioplastics showed high UV absorbance. This study highlights the potential of biodegradable plastics to pose enhanced ecotoxicological risk due to weathering under environmentally relevant conditions.

Chemicals sorbed to environmental microplastics are toxic to early life stages of aquatic organisms.

Microplastics are ubiquitous in aquatic ecosystems, but little information is currently available on the dangers and risks to living organisms. In order to assess the ecotoxicity of environmental microplastics (MPs), samples were collected from the beaches of two islands in the Guadeloupe archipelago, Petit-Bourg (PB) located on the main island of Guadeloupe and Marie-Galante (MG) on the second island of the archipelago. These samples have a similar polymer composition with mainly polyethylene (PE) and polypropylene (PP). However, these two samples are very dissimilar with regard to their contamination profile and their toxicity. MPs from MG contain more lead, cadmium and organochlorine compounds while those from PB have higher levels of copper, zinc and hydrocarbons. The leachates of these two samples of MPs induced sublethal effects on the growth of sea urchins and on the pulsation frequency of jellyfish ephyrae but not on the development of zebrafish embryos. The toxic effects are much more marked for samples from the PB site than those from the MG site. This work demonstrates that MPs can contain high levels of potentially bioavailable toxic substances that may represent a significant ecotoxicological risk, particularly for the early life stages of aquatic animals.

Towards standard methods for the classification of aquatic toxicity for biologically active household chemicals (BAHC) present in plastics, pharmaceuticals, and cosmetic products.

A standard method to test the aquatic toxicity of biologically active household chemicals (BAHC), including those with very low water solubility, is proposed. The method uses the common marine models Paracentrotus lividus embryos and Acartia clausi larvae, in order to advance towards derivation of water quality criteria for these emerging pollutants that currently lack environmental standards. Depending on the water solubility and octanol-water partition coefficient (Kow) of the substance, the protocol consists of testing the toxicity of the substances by serial dilutions of water stocks, dimethyl-sulfoxide stocks, or 100 mg/L lixiviates in seawater. When this method is applied to eleven model BAHC, the pharmaceutical fluoxetine, the antioxidant butylated hydroxytoluene, and the UV filters broadly present in cosmetics octocrylene and 4-methylbenzylidene camphor, are classified as very toxic to aquatic life, since their EC50 values are < 1 mg/L. In general, both biological models, P. lividus and A. clausi, yield the same classification of the substances tested, but variations in the classification of aquatic toxicity depending on methodological aspects are discussed. The use of A. clausi nauplii provides more protecting value to the toxicity parameters obtained by using this protocol.

In vivo oral bioavailability of Pb sequestered in metal rich granules in bivalves.

The present study was designed to evaluate in vivo the oral bioavailability of lead (Pb) present in the marine bivalve Dosinia exoleta. This infaunal clam, despite inhabiting in clean areas, presents Pb concentrations that are over the 1.5 mg kg-1 wet weight limit for human consumption set by the European Commission. However, Pb is accumulated in this clam in the form of metal rich granules, and it has been shown to be unavailable for trophic transfer to a marine decapod, so it was hypothesised that it might be unavailable for human consumers as well. Twelve Sprague Dawley rats were fed during 14 days with a diet including control mussels (Mytilus galloprovincialis), D. exoleta, or mussels enriched in Pb to the same levels as those found in D. exoleta. Pb accumulation in different rat tissues (blood, bone, kidneys and liver) was analysed. It was observed that Pb assimilation from D. exoleta was about half of Pb assimilation from M. galloprovincialis, and absolute bioavailabilities were around 2% for M. galloprovincialis and 1% for D. exoleta. These results suggest that it might be possible to increase the limit for human consumption for this bivalve to 3 mg kg-1 wet weight without representing an increase in the risk for consumers.

Shotgun Proteomics Analysis Discards Alkali Labile Phosphate as a Reliable Method To Assess Vitellogenin Levels in Mytilus galloprovincialis.

Vitellogenin, the egg yolk precursor, is a well-known biomarker of endocrine disruption in oviparous vertebrates. In invertebrates, such as bivalves, it has been used in the last 10 years for the same purpose, despite the limited knowledge of invertebrate endocrinology. In bivalves, vitellogenin levels are usually estimated using an indirect technique, alkali labile phosphate (ALP), that assumes that vitellogenin is the most abundant phosphorylated protein in the analyzed tissue. In this study, we applied shotgun proteomics for the identification and quantification of vitellogenin in marine mussel gonads and compared the results with those obtained with the ALP method. The proteomic analysis revealed that vitellogenin is only detected in female gonads with expression levels that are rather variable among female mussels at different stages of gonad development. ALP analysis, on the contrary, detected similar amounts of phosphorylated proteins regardless of sex or gonad development stage. These results show evidence that the ALP method is not providing reliable information about Vtg levels, at least in marine mussel gonads. ALP is not a good proxy to assess Vtg levels in marine mussels, and careful verification of the adequacy of the procedure should be done before ALP is further assumed as a proxy of Vtg in other bivalve mollusks.

Subcellular distribution and trophic transfer of Pb from bivalves to the common prawn Palaemon serratus.

The edible clam Dosinia exoleta has been reported to accumulate high contents of lead (Pb) in soft tissues disregarding the levels of Pb in the environment. This is due to the retention of Pb in the form of metal rich granules (MRG) in their kidneys throughout the mollusc lifespan. The potential for trophic transfer of Pb in this form to predators is expected to be low, since metals in the form of MRG are generally supposed to be trophically unavailable, but this assumption is based on studies with other metals (Ag, Cd, Cu or Zn) and has not been demonstrated with Pb until now. This study was designed to test if the Pb present in D. exoleta in the form of MRG is available to a decapod consumer, the common prawn Palaemon serratus, in comparison with a mussel diet showing a different subcellular distribution of Pb. As hypothesised, despite the high Pb concentrations (15µgg-1ww) offered to the prawns as D. exoleta tissues, Pb was almost completely unavailable for trophic transfer, and the prawns fed with this diet during 28 days showed the same Pb accumulation as prawns fed with a control diet with a much lower Pb concentration. On the contrary, individuals fed with mussel tissues containing the same Pb concentrations as the diet based on D. exoleta tissues showed 10 times higher Pb bioaccumulation, corresponding to a trophic transfer factor of 1.1%. Subcellular fractionation experiments revealed that the fraction of Pb in the form of MRG was much lower for the mussel, confirming, as observed for other metals, that MRG-associated Pb is not available for trophic transfer to decapod crustaceans.

Acute toxicity of selected organic pollutants to saltwater (mysid Siriella armata) and freshwater (cladoceran Daphnia magna) ecotoxicological models.

The acute toxicity of three organophosphate pesticides (diazinon, chlorpyrifos, pirimiphos-methyl) two chlorinated biocides (endosulfan, pentachlorophenol) and a nonsteroidal anti-inflammatory drug (diclofenac) was tested on <24 h neonates of saltwater mysidacea (Siriella armata) in laboratory acute toxicity test. The 50 and 10 % lethal effective concentrations (LC(50) and LC(10)), NOEC and LOEC values were calculated. The three organophosphate pesticides showed an apparently biphasic dose-response profile, supporting that its mechanism of action in crustaceans differ from other organic compounds. The biphasic pattern of response was confirmed using the common aquatic ecotoxicological model Daphnia magna. According to the 96-h LC(50) values for S. armata, the ranking of toxicity was chlorpyrifos (0.13 µg/L) < pirimiphos-methyl (1.3 µg/L) < endosulfan (3.2 µg/L) < diazinon (4.03 µg/L) < pentachlorophenol (262.2 µg/L) < diclofenac (2919 µg/L). In general, mysids resulted at least one order of magnitude more sensitive than model daphnia, which stresses the need for using marine species for the derivation of seawater quality standards.

Bioaccumulation of BDE-47 and effects on molecular biomarkers acetylcholinesterase, glutathione-S-transferase and glutathione peroxidase in Mytilus galloprovincialis mussels.

Mussels, Mytilus galloprovincialis, showed a high bioaccumulation ability when exposed to waterborne tetrabromodiphenyl ether (BDE-47), with a bioconcentration factor of 10,900 L Kg(-1) wet weight, and slow depuration rates in clean seawater. Kinetic and concentration-response experiments were performed measuring in the exposed mussel the activities of three molecular biomarkers: glutathione S-transferase (GST), glutathione peroxidase (GPx) and acetylcholinesterase (AChE). The long term (30 days) exposure of mussels to all concentrations (2-15 µg L(-1)) of BDE-47 significantly inhibited the AChE and GST activities, a result that supports the suitability of these biomarkers in marine pollution monitoring programs. However, GPx activity showed a less consistent pattern of response depending on the concentration and the duration of exposure.

Acute toxicity of tralopyril, capsaicin and triphenylborane pyridine to marine invertebrates.

A need for environmentally acceptable alternative antifouling (AF) biocides has arisen through restrictions in the use of many common biocides in the European Union through the Biocidal Product Regulation (Regulation EU No. 528/2012). Three such alternatives are triphenylborane pyridine (TPBP), tralopyril and capsaicin. This study aims at extending the available information on the toxicity of these three emerging AF biocides to key marine invertebrates. Here we investigate the toxicity of tralopyril and capsaicin to the early life stages of the mussel Mytilus galloprovincialis and the sea urchin Paracentrotus lividus and also of tralopyril, capsaicin and TPBP to the early life stages of the copepod Tisbe battagliai. The EC50 that causes abnormal development of mussel's D-veliger larvae and impairs the growth of sea urchin pluteus larvae are respectively 3.1 and 3.0 µg/L for tralopyril and 3,868 and 5,248 µg/L for capsaicin. Regarding the copepod T. battagliai, the LC50 was 0.9 µg/L for tralopyril, 1,252 µg/L for capsaicin and 14 µg/L for TPBP. The results obtained for the three substances are compared to a reference AF biocide, tributyltin (TBT), and their ecological risk evaluated. These compounds pose a lower environmental risk than TBT but still, our results suggest that tralopyril and TPBP may represent a considerable threat to the ecosystems.

Effects of primary leachates of conventional and alternative plastics in Cyprinodon variegatus fish larvae: Endocrine disruption and toxicological responses.

The inclusion of hazardous substances in the formulation of plastics raises significant concerns, particularly, if those substances are released as primary leachates during plastic degradation and/or fragmentation. In this sense, the production of degradable plastics holding deleterious additives can increase the release of harmful substances into the environment. Additionally, the effects of primary leachates of "eco-friendly" materials remain unexplored. To address this, we performed exposures to primary leachates of alternative polymers, and commercial bags to verify possible responses associated with endocrine disruption and/or activation of the detoxification pathway in larvae of the marine fish model Cyprinodon variegatus. The chemical characterization evidenced a great number of additives in the formulation of the materials analyzed in this study. Those include, except for the PLA sample, relevant levels of the hazardous phthalates DEHP and DiBP. Regarding the effects on marine fish larvae, exposure to leachates from alternative polymers (10 g/L) PHB and PHBV produced remarkable mortality (100%). While the exposure to bag leachates of all tested materials (1 and 10 g/L) produced alterations in biomarkers for steroidogenic and detoxification pathways. To a lesser extent (10 g/L), three materials produced significant alterations in estrogenic biomarkers (Home-compostable bag 1, LDPE and Recycled PE bags). Although the alterations in gene expression were not directly correlated to the amount of DEHP or DiBP, we can conclude that primary leachates of "eco-friendly" bags are harmful to marine vertebrates.

Marine degradation and ecotoxicity of conventional, recycled and compostable plastic bags.

Plastic bags are currently a major component of marine litter, causing aesthetical nuisance, and undesirable effects on marine fauna that ingest them or are entangled. Plastic litter also rises concern on the ecotoxicological effects due to the potential toxicity of the chemical additives leached in aquatic environments. Conventional plastic bags are made of polyethylene, either from first use or recycled, but regulations restricting single-use plastics and limiting lightweight carrier bags (<50 µm thickness) have fostered the replacement of thin PE bags by compostable materials advertised as safer for the environment. In this study, we assess the degradation of commercially available plastic bags in marine conditions at two scales: aquariums (60 days) and outdoors flow-through mesocosm (120 days). Strength at break point and other tensile strength parameters were used as ecologically relevant endpoints to track mechanical degradation. Ecotoxicity has been assessed along the incubation period using the sensitive Paracentrotus lividus embryo test. Whereas PE bags did not substantially lose their mechanical properties within the 60 d aquarium exposures, compostable bags showed remarkable weight loss and tensile strength decay, some of them fragmenting in the aquarium after 3-4 weeks. Sediment pore water inoculum promoted a more rapid degradation of compostable bags, while nutrient addition pattern did not affect the degradation rate. Longer-term mesocosms exposures supported these findings, as well as pointed out the influence of the microbial processes on the degradation efficiency of compostable/bioplastic bags. Compostable materials, in contrast toPE, showed moderate toxicity on sea-urchin larvae, partially associated to degradation of these materials, but the environmental implications of these findings remain to be assessed. These methods proved to be useful to classify plastic materials, according to their degradability in marine conditions, in a remarkably shorter time than current standard tests and promote new materials safer for the marine fauna.

Weathering increases the acute toxicity of plastic pellets leachates to sea-urchin larvae-a case study with environmental samples.

Microplastics, particles under 5 mm, pervade aquatic environments, notably in Tarragona's coastal region (NE Iberian Peninsula), hosting a major plastic production complex. To investigate weathering and yellowness impact on plastic pellets toxicity, sea-urchin embryo tests were conducted with pellets from three locations-near the source and at increasing distances. Strikingly, distant samples showed toxicity to invertebrate early stages, contrasting with innocuous results near the production site. Follow-up experiments highlighted the significance of weathering and yellowing in elevated pellet toxicity, with more weathered and colored pellets exhibiting toxicity. This research underscores the overlooked realm of plastic leachate impact on marine organisms while proposes that prolonged exposure of plastic pellets in the environment may lead to toxicity. Despite shedding light on potential chemical sorption as a toxicity source, further investigations are imperative to comprehend weathering, yellowing, and chemical accumulation in plastic particles.

A Practical Tool for the Assessment of Polymer Biodegradability in Marine Environments Guides the Development of Truly Biodegradable Plastics.

Environmental persistence is one of the few shortcomings of plastic materials. As a consequence, alternative plastics labeled as compostable are replacing polyolefins in some commercial applications, such as food bags and trash bags. A rapid, high-throughput, and environmentally relevant method to assess the potential biodegradability in marine conditions is used to assess these materials already on the market, as well as novel bio-based polymers still in development. By fitting experimental data to a non-linear logistic model, ultimate biodegradability can be calculated without regard for incubation time. Whereas the commercial products show negligible or very low marine biodegradability, one of the novel materials exceeds the 20% biodegradation threshold relative to fully marine biodegradable PHB after 28 days. In addition, the sensitivity of the method can be enhanced and its duration reduced, at the expense of labor-demanding preconditioning of the microbial inoculum, by increasing the bacterial density in the incubation vessels. In contrast, pre-exposure of the inoculum to plastic, either in laboratory or field conditions, does not enhance the performance of the test.

Filtration of biopolymer PHB particles loaded with synthetic musks does not cause significant bioaccumulation in marine mussels.

The role of the biopolymer polyhydroxybutyrate (PHB, <250 µm) as a vehicle of a synthetic musks mixture (celestolide, galaxolide, tonalide, musk xylene, musk moskene and musk ketone) to Mytilus galloprovincialis was investigated. For 30 days, virgin PHB, virgin PHB+musks (6.82 µg g-1) and weathered PHB+musks, were daily spiked into tanks containing mussels, followed by a 10-day depuration period. Water and tissues samples were collected to measure exposure concentrations and accumulation in tissues. Mussels were able to actively filter microplastics in suspension but the concentration of the musks found in tissues (celestolide, galaxolide, tonalide) were markedly lower than the spiked concentration. Estimated Trophic Transfer Factors suggest that PHB will only play a minor role on musks accumulation in marine mussels, even if our results suggest a slightly extended persistence in tissues of musks loaded to weathered PHB.

Evaluating the alterations of the estrogen-responsive genes in Cyprinodon variegatus larvae for biomonitoring the impacts of estrogenic endocrine disruptors (EEDs).

Currently, endocrine disruptors (EDs) can be found in all the environmental compartments. To understand the effects of estrogenic EDs (EEDs), adults of Cyprinodon variegatus have been classically used as a marine model. However, it is during development that exposure to contaminants may generate permanent consequences. Thus, the aim of this study was to verify the effects produced by acute exposure to 17α-ethinylestradiol (EE2) in C. variegatus larvae. Quantitative PCR (qPCR) results revealed the induction of vtg and zp gene expression on exposure to 1000 ng/L EE2 and the induction of vtgc, zp2, zp3 and cyp19a2, and inhibition of vtgab, wap and cyp1a1 on exposure to 100 ng/L EE2. Lower concentrations inhibited the gene expression of vtgab and wap (50 ng/L), cyp1a1 (25 ng/L) and zp2 (12.5 ng/L). These alterations in gene expression allow us to affirm that larvae of C. variegatus are an efficient and sensitive model for biomonitoring EEDs.

A protocol for lixiviation of micronized plastics for aquatic toxicity testing.

Plastics contain various types and amounts of additives that can leach into the water column when entering aquatic ecosystems. Some leached plastic additives are hazardous to marine biota at environmentally relevant concentrations. Disparate methodological approaches have been adopted for toxicity testing of plastic leachates, making comparison difficult. Here we propose a protocol to standardize the methodology to obtain leachates from microplastics (MPs) for aquatic toxicity testing. Literature reviewing and toxicity tests using marine model organisms and different types of MPs were conducted to define the main methodological aspects of the protocol. Acute exposure to leachates from the studied plastics caused negative effects on the early life stages of sea urchins and marine bacteria. We provide recommendations of key factors influencing lixiviation of MPs , such as particle size (<250 µm), solid-to-liquid ratio (1-10 g/L), mixing conditions (1-60 rpm), and lixiviation time (72 h). The proposed methodology was successful to determine the toxicity of leachates from different micronized plastics on marine biota. Our recommendations balance sensitivity, feasibility and environmental relevance, and their use would help ensure comparability amongst studies for a better assessment of the toxicity of plastic leachates on aquatic biota.

Occurrence and size distribution study of microplastics in household water from different cities in continental Spain and the Canary Islands.

The purpose of this study was to investigate the occurrence of microplastics (MPs) in drinking water in Spain by comparing tap water from different locations using common sampling and identification procedures. We sampled tap water from 24 points in 8 different locations from continental Spain and the Canary Islands by means of 25 µm opening size steel filters coupled to household connections. All particles were measured and spectroscopically characterized including not only MPs but also particles consisting of natural materials with evidence of industrial processing, such as dyed natural fibres, referred insofar as artificial particles (APs). The average concentration of MPs was 12.5 ± 4.9 MPs/m3 and that of anthropogenic particles 32.2 ± 12.5 APs/m3. The main synthetic polymers detected were polyamide, polyester, and polypropylene, with lower counts of other polymers including the biopolymer poly(lactic acid). Particle size and mass distributions were parameterized by means of power law distributions, which allowed performing estimations of the concentration of smaller particles provided the same scaling parameter of the power law applies. The calculated total mass concentration of the identified MPs was 45.5 ng/L. The observed size distribution of MPs allowed an estimation for the concentration of nanoplastics (< 1 µm) well below the ng/L range; higher concentrations are not consistent with scale invariant fractal fragmentation. Our findings showed that MPs in the drinking water sampled in this work do not represent a significant way of exposure to MPs and would probably pose a negligible risk for human health.

A nationwide monitoring of atmospheric microplastic deposition.

Plastic production continues to increase every year, yet it is widely acknowledged that a significant portion of this material ends up in ecosystems as microplastics (MPs). Among all the environmental compartments affected by MPs, the atmosphere remains the least well-known. Here, we conducted a one-year simultaneous monitoring of atmospheric MPs deposition in ten urban areas, each with different population sizes, economic activities, and climates. The objective was to assess the role of the atmosphere in the fate of MPs by conducting a nationwide quantification of atmospheric MP deposition. To achieve this, we deployed collectors in ten different urban areas across continental Spain and the Canary Islands. We implemented a systematic sampling methodology with rigorous quality control/quality assurance, along with particle-oriented identification and quantification of anthropogenic particle deposition, which included MPs and industrially processed natural fibres. Among the sampled MPs, polyester fibres were the most abundant, followed by acrylic polymers, polypropylene, and alkyd resins. Their equivalent sizes ranged from 22 µm to 398 µm, with a median value of 71 µm. The particle size distribution of MPs showed fewer large particles than expected from a three-dimensional fractal fragmentation pattern, which was attributed to the higher mobility of small particles, especially fibres. The atmospheric deposition rate of MPs ranged from 5.6 to 78.6 MPs m-2 day-1, with the higher values observed in densely populated areas such as Barcelona and Madrid. Additionally, we detected natural polymers, mostly cellulosic fibres with evidence of industrial processing, with a deposition rate ranging from 6.4 to 58.6 particles m-2 day-1. There was a positive correlation was found between the population of the study area and the median of atmospheric MP deposition, supporting the hypothesis that urban areas act as sources of atmospheric MPs. Our study presents a systematic methodology for monitoring atmospheric MP deposition.

Linking chemical contamination to biological effects in coastal pollution monitoring.

To establish the connection between pollutant levels and their harmful effects on living resources, coastal monitoring programmes have incorporated biological tools, such as the scope for growth (SFG) in marine mussels and benthic macrofauna community indices. Although the relation between oxygen-depleting anthropogenic inputs and the alteration of benthic communities is well described, the effects of chemical pollutants are unknown because they are not expected to favour any particular taxa. In this study, the combined efforts of five research teams involved in the investigative monitoring of marine pollution allowed the generation of a multiyear data set for Ría de Vigo (NW Iberian Peninsula). Multivariate analysis of these data allowed the identification of the chemical-matrix combinations responsible for most of the variability among sites and the construction of a chemical pollution index (CPI) that significantly (P < 0.01) correlated with biological effects at both the individual and the community levels. We report a consistent reduction in the physiological fitness of local populations of mussels as chemical pollution increases. The energy balance was more sensitive to pollution than individual physiological rates, but the reduction in the SFG was primarily due to significantly decreased clearance rates. We also found a decrease in benthic macrofauna diversity as chemical pollution increases. This diversity reduction resulted not from altered evenness, as the classic paradigm might suggest, but from a loss of species richness.

Prospective ecological risk assessment of sediment resuspension in an estuary.

This study assesses potential ecological risk of resuspended sediment in the water column during the construction of a viaduct in the estuary of the Ulla river (Galicia, NW Iberian Peninsula), a shellfish production area. Chemical analyses and toxicity bioassays with elutriates were performed with sediments from the area where the three pillars of the viaduct will be located (CT1, CT2 and CT3) and a reference sediment (A2). Acute toxicity of the elutriate was evaluated in five species of three trophic levels (Isochrysis galbana, Paracentrotus lividus, Mytilus galloprovincialis, Venerupis pullastra and Siriella armata). The sediments of the pillars showed moderate levels of contamination by trace elements (Cu, Cr). Clam and sea urchin embryo-larval toxicity tests showed slightly higher sensitivity than mussel embryo tests, and toxicity was not detected for phytoplankton and mysid bioassays. The predicted no-effect environmental concentration (PNEC) was calculated from the arithmetic mean of the lowest calculated EC(50)s for each sampling site. The predicted environmental concentration (PEC) was estimated from a simple dilution model and the PEC/PNEC ratio was calculated according to different scenarios of resuspension. Negligible ecological risk in the water column is expected during construction of the pillars.