Enzymatic Synthesis and Structural Modeling of Bio-Based Oligoesters as an Approach for the Fast Screening of Marine Biodegradation and Ecotoxicity.

Given the widespread use of esters and polyesters in products like cosmetics, fishing nets, lubricants and adhesives, whose specific application(s) may cause their dispersion in open environments, there is a critical need for stringent eco-design criteria based on biodegradability and ecotoxicity evidence. Our approach integrates experimental and computational methods based on short oligomers, offering a screening tool for the rapid identification of sustainable monomers and oligomers, with a special focus on bio-based alternates. We provide insights into the relationships between the chemical structure and properties of bio-based oligomers in terms of biodegradability in marine environments and toxicity in benchmark organisms. The experimental results reveal that the considered aromatic monomers (terephthalic acid and 2,5-furandicarboxylic acid) accumulate under the tested conditions (OECD 306), although some slight biodegradation is observable when the inoculum derives from sites affected by industrial and urban pollution, which suggests that ecosystems adapt to non-natural chemical pollutants. While clean seas are more susceptible to toxic chemical buildup, biotic catalytic activities offer promise for plastic pollution mitigation. Without prejudice to the fact that biodegradability inherently signifies a desirable trait in plastic products, nor that it automatically grants them a sustainable "license", this study is intended to facilitate the rational design of new polymers and materials on the basis of specific uses and applications.

Ecotoxicity of marine sediments: Sampling and laboratory artifacts and their impacts on risk classification.

Assessing the ecotoxicological risk of marine sediments is now a critical factor in deciding how to treat dredged material in harbor and coastal areas. Although ecotoxicological analyses are routinely required by some regulatory agencies in Europe, laboratory skills necessary to perform them are often underestimated. According to the Italian Ministerial decree No. 173/2016, ecotoxicological tests are performed on the solid phase and elutriates, and the classification of sediment quality is defined using the "Weight of Evidence" (WOE) approach. However, the decree does not provide adequate information regarding the preparation techniques and laboratory skills. As result, a wide variability among laboratories occurs. An error in the classification of ecotoxicological risk has a negative impact on the whole environmental quality and/or the economy and management of the interested area. Thus, the main aim of this study was to determine if such variability can affect the ecotoxicological outcomes of tested species and WOE associated classification, producing different options for the management of dredged sediments. Four different sediment types were selected to assess the ecotoxicological responses and their changes as a function of variability of the following factors: a) the storage time laps (STL) for both the solid phase and the elutriates; b) the methods used to prepare the elutriates (centrifugation vs. filtration), and the conservation method used for the elutriates (freshly prepared vs. freezing). Results suggest a wide variability of ecotoxicological responses among the four sediment samples here considered, differentiated according to chemical pollution, grain-size texture, and macronutrient contents. The storage time laps significantly affect the physicochemical parameters and the ecotoxicity of both the solid phase test and elutriates. For the elutriates preparation, centrifugation is preferred to filtration to preserve a better representation of sediment heterogeneity. Freezing of elutriates does not seem to show any significant effects on the toxicity. Findings allow to define a weighted schedule of the storage time of sediments and elutriates useful for laboratories to scale analytical priority and strategies related to different sediment types.

Navigating a Microplastic Sea: How the Pacific Cupped Oyster (Magallana gigas) Respond to Microplastic Pollution in Lagoons.

Microplastic pollution poses an escalating concern, particularly in coastal lagoons rich in biodiversity. This study delved into the occurrence of microplastics (MPs) in Magallana gigas (formerly Crassostrea gigas) from the Orbetello and Varano coastal lagoons (Italy), also investigating the response of these filter-feeding organisms to various colors (P = pink; B = blue; W = white) of high-density polyethylene (HDPE) MP fragments. Oysters were exposed for 7 days under controlled conditions. Subsequently, the oysters underwent analysis for both MP presence and biochemical markers of oxidative stress. Diverse ingestion rates of HDPE were noted among oysters from the two lagoons, eliciting antioxidant responses and modifying baseline activity. The two-way ANOVA revealed the significant effects of treatment (control; HDPE_B; HDPE_P; HDPE_W), site, and the interaction between treatment and site on all biomarkers. Non-metric multidimensional scaling showed a divergent effect of HDPE color on biomarkers. Further investigation is warranted to elucidate the mechanisms underlying the influence of MP color, dose-dependent effects, and the long-term impacts of exposure. Comprehending these intricacies is imperative for devising effective strategies to mitigate plastic pollution and safeguard marine health.

Chemical pollution and ecotoxicological effects of high-density polyethylene microplastics in Mytilus galloprovincialis from two Italian lagoon ecosystems.

Transitional water ecosystems have low water exchanges and can trap chemicals and microplastics (MPs). In this study, MPs, trace elements, polycyclic aromatic hydrocarbon-PHAs levels and the oxidative stress response were assessed in Mytilus galloprovincialis from two Italian lagoon ecosystems (Orbetello and Varano). In addition, the ecotoxicological effects induced by the exposure of M. galloprovincialis to high-density polyethylene-HDPE MPs were also determined. Levels of trace elements were almost always comparable among the sites, whereas MPs were found only in mussels from Orbetello. PAHs were always under the limit of quantification. Glutathione peroxidase and malondialdehyde levels were significantly higher in mussels from Varano. As regard the exposure test, it was found a significant effect of treatment, site and their interaction on mortality and biochemical biomarkers in both fed and unfed mussels. However, principal component analysis suggests similar effects of both color and nourishment condition on biochemical biomarkers. These findings warrant further investigation.

Ecotoxicological Assessment of "Glitter" Leachates in Aquatic Ecosystems: An Integrated Approach.

The most worrisome fraction within plastic pollution is that of microplastics (MP). A category of MP almost completely ignored is that of glitter. The objective of this study is to test the toxicity of nine types of glitter leachate (3 soak times: 3, 90 and 180 days) on model organisms in freshwater (Allivibrio fischeri, Raphidocelis subcapitata, Daphnia magna) and saltwater (Allivibrio fischeri, Phaeodactylum tricornutum, Paracentrotus lividus). An integrated approach was applied to obtain the percentage of ecotoxicological risk. The results show that (i) photosynthesizing primary producers are the most sensitive trophic level; (ii) algae transitioned from growth inhibition to biostimulation; (iii) D. magna showed higher sensitivity after 48 h compared to 24 h; (iv) A. fischeri responded more strongly in saltwater than in freshwater. The integrated data show a greater risk associated with the marine environment, with the highest risk for glitters that are hexagonal and composed of poly-methyl-methacrylate. Our multivariate analysis shows that the toxicity of plastic leaching is a complex phenomenon that depends on the sensitivity of the species, in some cases on the soaking time and on the medium, and is not clearly linked to the polymer type, the contact area or the colors of the particles.

Sparkling plastic: Effects of exposure to glitter on the Mediterranean mussel Mytilus galloprovincialis.

Microbeads and fragments have been widely studied, while glitter remains neglected by the literature although found in a variety product (e.g., body paints, nail polish, cosmetics, craft products). The main aim of this study was to assess the effects of different types and concentrations of glitter particles on Mytilus galloprovincialis after 7 days of exposure. The experiment was divided into a preliminary test and a confirmatory test. Our findings support the hypothesis for a link between concentration and type of glitter particles, percentage of recovery and oxidative stress in M. galloprovincialis. There was a significant correlation between particle length and percentage of particles recovered in water, suggesting that the digestive tract of M. galloprovincialis retains smaller particles more. In addition, we noted an increase in antioxidant defense induced by smaller particles. Moreover, certain types of glitter crumbled and shortened in length, resulting in higher levels of oxidative stress biomarkers. Finally, the star-shaped glitter particles had a different effect on oxidative stress biomarkers. Further studies are needed to clarify the toxic effects of glitter on aquatic organisms and to quantify its proportion to other microplastics in the environment.

2,5-Furandicarboxaldehyde as a bio-based crosslinking agent replacing glutaraldehyde for covalent enzyme immobilization.

In the quest for a bio-based and safer substitute for glutaraldehyde, we have investigated 2,5 diformylfuran (DFF) as bifunctional crosslinking agent for the covalent immobilization of glucoamylase on amino-functionalized methacrylic resins. Immobilization experiments and systematic comparison with glutaraldehyde at four different concentrations for the activation step showed that DFF leads to comparable enzymatic activities at all tested concentrations. Continuous flow experiment confirms a similar long term stability of the immobilized formulations obtained with the two crosslinkers. The NMR study of DFF in aqueous solution evidenced a much simpler behaviour as compared to glutaraldehyde, since no enolic forms can form and only a mono-hydrated form was observed. Unlike in the case of glutaraldehyde, DFF reacts covalently with the primary amino groups via imine bond formation only. Nevertheless, the stability of the covalent immobilization was confirmed also at acidic pH (4.5), most probably because of the higher stability of the imine bonds formed with the aromatic aldehydes. In terms of toxicity DFF has the advantage of being poorly soluble in water and, more importantly, poorly volatile as compared to glutaraldehyde, which displays severe respiratory toxicity. We have performed preliminary ecotoxicity assays using Aliivibrio fischeri, a marine bacterium, evidencing comparable behaviour (below the toxicity threshold) for both dialdehydes at the tested concentrations.

Lethal and Sublethal Responses of Hydropsyche pellucidula (Insecta, Trichoptera) to Commercial Polypropylene Microplastics after Different Preconditioning Treatments.

Microplastics (MPs) pose biological and chemical hazards in aquatic and terrestrial food webs across the globe. Research on microplastic contamination has long focused on marine ecosystems, whereas the toxicological impact on freshwater organisms is still little explored. In this study, the lethal and sublethal response of the freshwater macroinvertebrate Hydropsyche pellucidula exposed to polypropylene MPs after different pre-conditioning treatments was assessed. Field samples were collected in a riverine system (Vipacco river; northeast Italy) to assess the characteristics of the MPs in the aquatic environment Both water and sediment were contaminated by MPs (3.73 ± 2.11 items m-3 per min and 3.33 ± 4.16 items dm-3, respectively). The chemical MPs composition included polystyrene, polyethylene terephthalate, polyurethane, polyamide, polypropylene, and polyethylene. Polypropylene (PP), although not the most abundant polymer recorded in the study area, was preferred over the other types according to its abundance in freshwater and H. pellucidula feeding behavior. A housing test was performed to recreate the natural conditions of larvae sampled for a reliable response to the ecotoxicological tests. The microplastics underwent either preconditioning with Vipacco River water (PP-river) and surfactant Triton X-100 (PP-sf) or no pre-treatment (PP). Submersion of microplastics in 10 µg L-1 of surfactant solution for 24 h was sufficient to induce consistent spectral changes and modify the chemical profile of the plastic surface. Mortality rate differed according to treatment: PP and PP-river > positive control > PP-sf > negative control. Integrated biomarker response (IBRv2) and analysis of oxidative stress biomarker levels showed a greater response of superoxide dismutase and lipid peroxidation (malondialdehyde) in larvae treated with PP conditioned in surfactant. Our findings enhance knowledge on the toxicity of PP and conditioning phases on H. pellucidula larvae.

Exposure to pet-made microplastics: Particle size and pH effects on biomolecular responses in mussels.

This study aims to evaluate the expression of biomarkers of oxidative stress (LPO, GPx, AtCh, SOD) in mussels (Mytilus galloprovincialis) following the exposure to suspensions of microparticles irregular shaped fibres of Polyethylene terephthalate of different sizes (small 5-60 µm, S-PET; medium 61-499 µm, M-PET; large 500-3000 µm, L-PET) at a single dose of 0.1 g/L. Mussels were tested under two different starting pH conditions of marine water: standard (8.0) and acidified (7.5). The results obtained from this study show that: i) PET microplastics are able to induce biochemical stress in mussels; ii) among the biomarkers tested, LPO and GPx were more effective in detecting the stress induced by microplastic in both initial pH conditions; iii) the expression of biomarkers was influenced by the size of the microparticle. In particular, greater effects were associated with the largest PET particle tested (0.5-3.0 mm); iv) regarding the effect of pH, in experiments starting from 7.5 pH the animals showed a lower biomarker expression than those starting from 8.0 pH.

PET microplastics toxicity on marine key species is influenced by pH, particle size and food variations.

This study aims to evaluate effects induced by the exposure of key marine species to leachates and suspensions of different particle-size of PET microparticles, a plastic polymer that is actually considered safe for the environment. Leachates and suspensions of small (5-60 µm); medium (61-499 µm) and large (500-3000 µm) PET were tested on bacteria (V. fischeri; UNI EN ISO 11348-3:2009), algae (P. tricornutum; UNI EN ISO 10253:2016E), and echinoderms (P. lividus; EPA 600/R-95-136/Section 15) species both under standard (8.0) and acidified (7.5) pH conditions. Results obtained show that: i) conversely to larval stage of P. lividus, bacterial and algal tested species are not sensitive to PET pollution under all tested conditions; ii) different tested particle-sizes of PET produce effects that are not always related to their particle-size; iii) differences comparing acidified and standard pH conditions were recorded; iv) concerning echinoderms, food availability produce significant differences compared to fasting conditions; v) special attention on the possible interactions between MPs and other stressors (e.g., food and pH) is needed in order to give a better picture of natural occurring dynamics on marine ecosystems especially in the future frame of global changes.

Litter in alien species of possible commercial interest: The blue crab (Callinectes sapidus Rathbun, 1896) as case study.

Marine litter levels were measured in the stomach contents, hepatopancreas, and gonad tissues of crustacea decapod (Callinectes sapidus Rathbun, 1896; n = 6), a widespread alien species affecting the Lesina lagoon. Results highlight a mean of 2.5 (SD = 1.6) items/animal and indicate the presence of metals fragments (13%) and plastics (13% PE; 6.7% PET) in the stomach contents of collected individuals. No microplastic particles were detected in the hepatopancreas or in muscle tissue, while microplastic fibres (nylon, rayon, polyester) were found present in female specimen gonads. The presence of synthetic fibres in the investigated species reflects the relative contamination level in this habitat type and suggests that the blue crab could be considered a model organism for evaluating the contamination status of the study area.

Mediterranean Coastal Lagoons: The Importance of Monitoring in Sediments the Biochemical Composition of Organic Matter.

Transitional water ecosystems are targeted by the European Union (EU) Water Framework Directive (WFD, CE 2000/60) monitoring programs in coastal zones. Concerning sediments, activities performed for the WFD focus on a few variables concerning the biochemical composition of organic matter. Our research reports the effects of oxygen availability on the biochemical composition of organic matter in sediments to highlight levels of targeted variables in time and, according to the depth of sediment layer, both under oxygenated and anoxic conditions in a mesocosm study on sediment cores. Results provide evidence that tested factors of interest (i.e., disturbance type, oxygenic versus anoxic conditions; persistence time of disturbance, 0-14 days; penetration through sedimentary layers, 0-10 cm depth) are able to significantly affect the biochemical composition of organic matter in sediments. Large part of the variables considered in this study (total organic carbon (TOC), total phosphorous (TP), total sulphur (TS), Fe, carbohydrates (CHO), total proteins (PRT), biopolymeric carbon (BPC), chlorophyll-a (Chl-a) are significantly affected and correlated to the oxygenation levels and could be good early indicators of important changes of environmental conditions. Monitoring activities performed under WFD guidelines and management strategies of Mediterranean coastal lagoon ecosystems shall include the biochemical composition of organic matter in sediment to provide an exhaustive picture of such dynamic ecosystems.