Assessing combined effects of long-term exposure to copper and marine heatwaves on the reef-forming serpulid Ficopomatus enigmaticus through a biomarker approach.

Sessile benthic organisms can be affected by global changes and local pressures, such as metal pollution, that can lead to damages at different levels of biological organization. Effects of exposure to marine heatwaves (MHWs) alone and in combination with environmentally relevant concentration of copper (Cu) were evaluated in the reef-forming tubeworm Ficopomatus enigmaticus using a multi-biomarker approach. Biomarkers of cell membrane damage, enzymatic antioxidant defences, metabolic activity, neurotoxicity, and DNA integrity were analyzed. The exposure to Cu alone did not produce any significant effect. Exposure to MHWs alone produced effects only on metabolic activity (increase of glutathione S-transferase) and energy reserves (decrease in protein content). MHWs in combination with copper was the condition that most influenced the status of cell homeostasis of exposed F. enigmaticus. The combination of MHWs plus Cu exposure induced increase of protein carbonylation and glutathione S-transferase activity, decrease in protein/carbohydrate content and carboxylesterase activity. This study on a reef-forming organism highlighted the additive effect of a climate change-related stressor to metals pollution of marine and brackish waters.

Ocean acidification impairs seagrass performance under thermal stress in shallow and deep water.

Despite the effects of ocean acidification (OA) on seagrasses have been widely investigated, predictions of seagrass performance under future climates need to consider multiple environmental factors. Here, we performed a mesocosm study to assess the effects of OA on shallow and deep Posidonia oceanica plants. The experiment was run in 2021 and repeated in 2022, a year characterized by a prolonged warm water event, to test how the effects of OA on plants are modulated by thermal stress. The response of P. oceanica to experimental conditions was investigated at different levels of biological organization. Under average seawater temperature, there were no effects of OA in both shallow and deep plants, indicating that P. oceanica is not limited by current inorganic carbon concentration, regardless of light availability. In contrast, under thermal stress, exposure of plants to OA increased lipid peroxidation and decreased photosynthetic performance, with deep plants displaying higher levels of heat stress, as indicated by the over-expression of stress-related genes and the activation of antioxidant systems. In addition, warming reduced plant growth, regardless of seawater CO2 and light levels, suggesting that thermal stress may play a fundamental role in the future development of seagrass meadows. Our results suggest that OA may exacerbate the negative effects of future warming on seagrasses.

Robinia pseudoacacia L. (Black Locust) Leaflets as Biomonitors of Airborne Microplastics.

Here we investigate the suitability of Robinia pseudoacacia L. (black locust) leaflets as a novel biomonitor of airborne microplastics (MPs) including tyre wear particles (TWPs). Leaflets were collected from rural roadside locations (ROs, n = 5) and urban parks (UPs, n = 5) in Siena, Italy. MPs were removed by washing, identified by stereomicroscope, and analysed for polymer type by Fourier transform infrared spectroscopy. Daily MP deposition was estimated from leaf area. The mass magnetic susceptibility and the bioaccumulation of traffic-related potentially toxic elements (PTEs) were also analysed. The total number of MPs at ROs was significantly higher at 2962, dominated by TWPs, compared with 193 in UPs, where TWPs were not found. In contrast, total microfibres were significantly higher in UPs compared with ROs (185 vs. 86). Daily MP deposition was estimated to range from 4.2 to 5.1 MPs/m2/d across UPs and 29.9-457.6 MPs/m2/d across ROs. The polymer types at ROs were dominated by rubber (80%) from TWPs, followed by 15% polyamide (PA) and 5% polysulfone (PES), while in UPs the proportion of PES (44%) was higher than PA (22%) and polyacrylonitrile (11%). The mean mass magnetic susceptibility, a proxy of the bioaccumulation of traffic-related metallic particles, was higher at ROs (0.62 ± 0.01 10-8 m3/kg) than at UPs (-0.50 ± 0.03 10-8 m3/kg). The content of PTEs was similar across sites, except for significantly higher concentrations of Sb, a tracer of vehicle brake wear, at ROs (0.308 ± 0.008 µg/g) compared with UPs (0.054 ± 0.006 µg/g). Our results suggest that the waxy leaflets and easy determination of surface area make Robinia an effective biomonitor for airborne MPs including TWPs.

Impact of global change on environmental hazards of different clays: A case study on Aliivibrio fischeri.

The effects of global change in marine ecosystems are expected to lower pH from the current 8.1-7.5-7.0, which will have significant impacts on marine species. The purpose of this study is to investigate whether the ecotoxicity of ten different natural clays change significantly in response to the acidification process and what factors are associated with the observed changes. In this study, the ecotoxicological response of a bacterium (Aliivibrio fischeri) was tested under current (pH= 8.1) and acidified (pH 7.5 and 7.0) conditions. The ecotoxicity detected in the solid phase test (SPT protocol) and in the contact water was affected by the pH, which increased the ecotoxicity from 2/10 clays (pH 8.10) to 7/10 clays (pH 7.00), also shifting the detected effects from low to high toxicity values. The analyses performed on the natural clays studied show that pH can affect the release of metals, metalloids and rare earths from the clays into the contact water phase, affecting the toxicity observed. This phenomenon depends on the type of clay and is closely related to its mineralogical composition. As consequence, in a globally changing scenario, ecotoxicity, even of natural materials such as clay, cannot be considered stable, but must be accurately revaluated depending on the mineralogical and chemical composition of the clay. Moreover, the mineralogical composition of clays showed different efficiency in absorbing bacteria on the surface of clay particles. It was found that live bacterial cells were absorbed on the clay surface in numbers that were dependent on both clay types and pH levels.

Multiple approach for assessing lagoon environmental status based on water bodies quality indices and microplastics accumulation.

Lagoon environments, like all the marine-coastal areas, offer a wide variety of ecosystem services, but at the same time are affected by pressing human activities that lead to deterioration of the environmental quality, loss of biodiversity, habitat destruction and pollution. Since the well-being of population and local economy depend on the environmental status of these ecosystems, it is essential to adopt long-term management tools to achieve the Good Environmental Status sensu European Marine Strategy Framework Directive and Water Framework Directive. A Nature 2000 site (Lesina lagoon, south Italy) was assessed within a project aimed at protecting and restoring biodiversity and lagoon habitat through integrated monitoring, suitable management, and good practices. Here we provide an assessment of the lagoon integrity based on a multi-metrics approach, highlighting match/mismatch among environmental quality indicators and microplastics (MP) pollution. Some environmental quality indices based on ecosystem components as vegetation, macroinvertebrates and water trophic variables were applied together with an accurate evaluation of MPs abundance, distribution, and composition to assess the ecological status of Lesina lagoon before and after cleaning actions with litter removal. Overall, all the ecological descriptors outlined a clear lagoon spatial gradient, with a western saltier and organic-enriched area characterized by the absence of vegetation, macrozoobenthos with lower diversity and richness and high MPs occurrence. The focus on macrozoobenthos, considered as a key component of the lagoon ecosystem, identified much more sites in "poor" status than the other indicators here considered. Moreover, it was found a negative relationship between the Multivariate Marine Biotic Index and MPs items in sediment, suggesting that MP pollution has a negative impact on macrobenthic fauna, concurring to the deterioration of the benthic ecological status.

Understanding Marine Biodegradation of Bio-Based Oligoesters and Plasticizers.

The study reports the enzymatic synthesis of bio-based oligoesters and chemo-enzymatic processes for obtaining epoxidized bioplasticizers and biolubricants starting from cardoon seed oil. All of the molecules had MW below 1000 g mol-1 and were analyzed in terms of marine biodegradation. The data shed light on the effects of the chemical structure, chemical bond lability, thermal behavior, and water solubility on biodegradation. Moreover, the analysis of the biodegradation of the building blocks that constituted the different bio-based products allowed us to distinguish between different chemical and physicochemical factors. These hints are of major importance for the rational eco-design of new benign bio-based products. Overall, the high lability of ester bonds was confirmed, along with the negligible effect of the presence of epoxy rings on triglyceride structures. The biodegradation data clearly indicated that the monomers/building blocks undergo a much slower process of abiotic or biotic transformations, potentially leading to accumulation. Therefore, the simple analysis of the erosion, hydrolysis, or visual/chemical disappearance of the chemical products or plastic is not sufficient, but ecotoxicity studies on the effects of such small molecules are of major importance. The use of natural feedstocks, such as vegetable seed oils and their derivatives, allows the minimization of these risks, because microorganisms have evolved enzymes and metabolic pathways for processing such natural molecules.

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.

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.

Interdisciplinary approach to solve unusual mortalities in the European common frog (Rana temporaria) in two high-mountain ponds affected by climate change.

The global decline in amphibian populations is a major environmental issue. Chytridiomycosis, Ranaviruses and the red-leg syndrome have been identified in unusual mortality events. However, these infections do not account for all causes of declining amphibian populations. Moreover, several cases of amphibian mortality are difficult to solve without resorting to an interdisciplinary approach. Two cases of unusual mortality in Rana temporaria occurred at two high-mountain ponds (northwest Italy) in April and May 2021. Water and frog samples were analysed to understand the possible causes responsible for the unusual mortalities. Results of the main physicochemical (pH, conductivity, dissolved oxygen, chemical and biochemical oxygen demand) and nutrient (ammonia/ammonium, nitrite, nitrate, total phosphorus) parameters revealed a good condition of the water quality, with the absence of the main cyanotoxins (microcystins/nodularins). However, unseasonably high spring water temperatures were recorded in both ponds (12.73 °C and 14.21 °C for Frog Pond and Selleries Pond, respectively). Frogs (n = 50; snout-vent length: 7.0-9.8 cm; body mass: 85-123 g) collected from Frog Pond mainly presented bumps on the ventral cavity and dermal ulceration associated with the isolation of Carnobacterium maltaromaticum. On the other hand, frogs (n = 5; snout-vent length: 8.0-9.1 cm; body mass: 87-92 g) from Selleries Pond presented petechiae and dermal ulcerations on the rear limbs associated with the isolation of Aeromonas salmonicida and A. sobria. In both mortality events, the interdisciplinary approach revealed an association between frog mortalities and the isolation of bacteria. Isolated bacteria are considered opportunistic pathogens, and the high values of the water temperature has certainly led a stress on the frogs, favouring the spread of bacteria and the death of the frogs. Further studies are needed to assess the pathophysiological effects of the opportunistic bacteria here isolated, clarifying the interactions between emerging pathogens and climate change.

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.

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.

Natural Active Ingredients for Poly (Lactic Acid)-Based Materials: State of the Art and Perspectives.

This review describes the state of the art in the field of poly (lactic acid) (PLA)-based materials activated by natural compounds and extracts (active ingredients, AIs) from plant sources for food and biomedical applications. With a multidisciplinary approach, after a description of the synthesis and properties of PLA, special attention was paid to the chemical properties and unconventional extraction technologies of AIs used for PLA activation. Innovative techniques for the incorporation of AIs into PLA; characterization and the antioxidant and antimicrobial properties of the novel materials were discussed. In view of future perspectives, this study has evidenced that some aspects need to be further investigated from joint research between academia and industry, according to the green chemistry principles and circular economy strategy.

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.

Multivariate tools to investigate the spatial contaminant distribution in a highly anthropized area (Gulf of Naples, Italy).

The Gulf of Naples located in a high anthropized coastal area is subjected to an infrastructural intervention for the installation of a submarine power pipeline. In order to evaluate the distribution of contaminants in the seafloor sediments, a preliminary study has been conducted in the area using multivariate techniques. The statistic approach was performed to gain insights on the occurrence of organic and inorganic contaminants within the area, aiming to identify the relevant hot spots. Three geographical sub-areas influenced by different contaminant association were recognized: Torre Annunziata (TA), Capri (CA), and middle offshore (MO). TA and CA resulted marked by a severe contamination pattern due to anthropogenic pressures. In addition, the influence of the depositional basin in governing the contamination trend has been pointed out. The supervised technique PLS_DA resulted to be a powerful tool in addressing the complexity of the huge dataset acquired during the marine survey, highlighting the main trends in the variability of quality indicators, orienting thus the deeper investigations during follow-up monitoring activities.

Polyethylene microplastics reduce filtration and respiration rates in the Mediterranean sponge Petrosia ficiformis.

Microplastic (MP) pollution represents a distinctive mark of the Anthropocene. Despite the increasing efforts to determine the ecological impacts of MP on marine biodiversity, our understanding of their toxicological effects on invertebrate species is still limited. Despite their key functional roles, sponges (Phylum Porifera) are particularly understudied in MP research. These filter-feeders extract and retain particles from the water column, across a broad size range. In this study, we carried out a laboratory experiment to assess the uptake of MPs (polyethylene, PE) by the Mediterranean sponge Petrosia ficiformis, how MPs influence key biological process after different times of exposure (24h and 72h) and whether they can be subsequently eliminated. MP uptake increased with time of exposure, with 30.6% of the inoculated MP particles found in sponge samples after 72h. MPs impaired filtration and respiration rates and these effects were still evident 72h after sponges had been transferred in uncontaminated water. Our study shows that time of exposure represents a key factor in determining MP toxicity in sponges. In addition, our results suggest that sponges are able to incorporate foreign particles and may thus be a potential bioindicator for MP pollutants.

Modulation of Antioxidant Defense in Farmed Rainbow Trout (Oncorhynchus mykiss) Fed with a Diet Supplemented by the Waste Derived from the Supercritical Fluid Extraction of Basil (Ocimum basilicum).

Phytotherapy is based on the use of plants to prevent or treat human and animal diseases. Recently, the use of essential oils and polyphenol-enriched extracts is also rapidly increasing in the aquaculture sector as a means of greater industrial and environmental sustainability. Previous studies assessed the antibacterial and antiparasitic effects of these bioactive compounds on fish. However, studies on the modulation of oxidative stress biomarkers are still scant to date. Thus, in this study, the modulation of antioxidant defense against oxidative stress exerted by fish diets supplemented with a basil supercritical extract (F1-BEO) was assessed in rainbow trout Oncorhynchus mykiss. The F1-BEO extracted with supercritical fluid extraction was added to the commercial feed flour (0.5, 1, 2, 3% w/w) and mixed with fish oil to obtain a suitable compound for pellet preparation. Fish were fed for 30 days. The levels of stress biomarkers such as superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, glutathione reductase, glyoxalase I, glyoxalase II, lactate dehydrogenase, glutathione and malondialdehyde showed a boost in the antioxidant pathway in fish fed with a 0.5% F1-BEO-supplemented diet. Higher F1-BEO supplementation led to a failure of activity of several enzymes and the depletion of glutathione levels. Malondialdehyde concentration suggests a sufficient oxidative stress defense against lipid peroxidation in all experimental groups, except for a 3% F1-BEO-supplemented diet (liver 168.87 ± 38.79 nmol/mg prot; kidney 146.86 ± 23.28 nmol/mg prot), compared to control (liver 127.76 ± 18.15 nmol/mg prot; kidney 98.68 ± 15.65 nmol/mg prot). Our results suggest supplementing F1-BEO in fish diets up to 0.5% to avoid potential oxidative pressure in farmed trout.

Occurrence of microplastics in the gastrointestinal tract of benthic by-catches from an eastern Mediterranean deep-sea environment.

Concern about microplastic pollution little is known about levels in deep-sea species; to fill this knowledge gap, levels of microplastics in the gastrointestinal (GI) tracts of 34 fish from eight different deep-sea by-catches: blackmouth catshark, lesser spotted dogfish, and velvet belly, armless snake eel, hollowsnout grenadier, phaeton dragonet, royal flagfin, and slender snipe eel were measured. All were collected at the same site (east Sardinia, Mediterranean Sea; 40°10'12.49″N, 9°44'12.31″E) using a bottom gillnet at depths between -820/250 and -1148 ft./350 m. Microplastics (MPs) were retrieved in 16 out of 34 fish. At least one microplastic item was found in 48% (33%, E. spinax - 75%, G. melastomus) of the samples. The most frequent was polyethylene (PE), with nine items (filaments, films, fragments) found in five specimens. This preliminary study of by-catches adds new data on MPs ingestion by species inhabiting a deep-sea environment of the Mediterranean.

Microplastics accumulation in functional feeding guilds and functional habit groups of freshwater macrobenthic invertebrates: Novel insights in a riverine ecosystem.

Microplastics pose a major threat for aquatic ecosystems, but the contamination dynamics in organisms inhabiting freshwater ecosystems is still little studied. Largely used for biomonitoring, macrobenthic invertebrates provide a pivotal trophic resource for many fish and bird species. In this study, we investigated the microplastics contamination in a macrobenthic invertebrate community (2772 individuals belonging to 33 taxa identified) in a high-plain riverine ecosystem (Vipacco River, northeast Italy) and compared the amount of microplastics accumulated in functional feeding guilds/functional habit groups. Microplastics (cellulosic fibers associated with polyester) were found in 48.5% of the taxa, with the highest amount detected in the collector-gatherers, followed by predators. The collector-gatherers showed a significantly higher microplastic accumulation than the other functional feeding guilds, whereas there was no difference among the functional habit groups. The main source of microplastics pollution was most likely urban wastewater discharge points located along the river. Our study reports a novel approach about microplastic pollution assessment in lotic environments, as it focuses into the microplastic contamination dynamics in an entire macrobenthic invertebrate community perspective and underlines the need for further study.

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.

A Multidisciplinary Approach to Evaluate the Effects of Contaminants of Emerging Concern on Natural Freshwater and Brackish Water Phytoplankton Communities.

Ecotoxicological assays on monospecific phytoplankton have limited application for detecting the effects of environmental pollutants on multiple species communities. With this study, we took an ecotoxicological, ecological, and biochemical approach to evaluate the effects of two contaminants of emerging concern (zinc oxide nanoparticles, ZnO NPs, and potassium dichromate, K2Cr2O7) at different concentrations (K2Cr2O7 5.6-18-50 mg/L; ZnO NPs 10-100-300 mg/L) on natural freshwater and brackish water phytoplankton communities. Cell density and absorbance values decreased in freshwater and brackish water phytoplankton communities after exposure to ZnO NPs (100 mg/L and 300 mg/L only for freshwater), whereas growth rate was increased in both freshwater and brackish water phytoplankton communities after exposure to ZnO NPs 10 mg/L. Differently, there was no clear relationship between concentration and inhibition growth after exposure to K2Cr2O7: the lowest cell density was recorded after exposure to 18 mg/L. Moreover, the evenness index value was lower compared to the other concentrations, indicating the growth of a few, albeit resistant species to higher K2Cr2O7 concentrations. Generally, Bacillariophyceae and Dinoficee were prevalent in phytoplankton cultures after exposure to ZnO NPs and K2Cr2O7. The Shannon-Wiener index was slightly higher in the negative than the positive controls, but diversity was low after all treatments in both ecotoxicological assays. The evenness index was always very close to zero, indicating the numerical predominance of one or very few species. Finally, the decrease in chlorophyll-a and pheophytin-a in both ecotoxicological assays indicated a change in photosynthetic activity. Our findings provide evidence for alterations in natural phytoplankton after exposure to emerging contaminants that can disrupt an entire ecosystem's integrity.