Feeding exposure and feeding behaviour as relevant approaches in the assessment of the effects of micro(nano)plastics to early life stages of amphibians.

The sedimentation of micro and nanosized plastics is of considerable environmental relevance and the need to assess its sublethal effects to biota increasingly recognized. In their majority, as bottom, non-selective grazers, independent-feeding young life stages of amphibians, an already severely endangered worldwide group, may be particularly vulnerable to sedimented plastics. Alongside, they may be good model organisms for the assessment of the effects of micro(nano)plastics (MNPs) through ingestion. However, to our knowledge, few studies have assessed amphibians' exposure to MNPs through contaminated food or its effects in feeding behaviour assays. The available studies reveal a lack of consistent methodology: organisms, food type, media of exposure, or exposure conditions (temperature and light) in the assessment of effects. This perspective article, will address major differences found in the available studies, identifying type, size and concentrations of the polymers tested, species, and observed effects, aiming to highlight the importance of feeding exposure assays when attempting to evaluate the effect of MNPs in amphibians.

The effects of nanoplastics on marine plankton: A case study with polymethylmethacrylate.

Marine biota is currently exposed to plastic pollution. The biological effects of plastics may vary according to polymer types (e.g. polystyrene, polyethylene, acrylate), size of particles (macro, micro or nanoparticles) and their shape. There is a considerable lack of knowledge in terms of effects of nanoplastics (NP) to marine biota particularly of polymers like polymethylmethacrylate (PMMA). Thus, this study aimed to assess its ecotoxicological effects using a battery of standard monospecific bioassays with four marine microalgae (Tetraselmis chuii, Nannochloropsis gaditana, Isochrysis galbana and Thalassiosira weissflogii) and a marine rotifer species (Brachionus plicatilis). The tested PMMA-NP concentrations allowed the estimation of median effect concentrations for all microalgae species. T. weissflogii and T. chuii were respectively the most sensitive (EC50,96h of 83.75 mg/L) and least sensitive species (EC50,96h of 132.52 mg/L). The PMMA-NP were also able to induce mortality in rotifers at concentrations higher than 4.69 mg/L with an estimated 48 h median lethal concentration of 13.27 mg/L. A species sensitivity distribution curve (SSD), constructed based on data available in the literature and the data obtained in this study, reveal that PMMA-NP appears as less harmful to marine biota than other polymers like polystyrene.

Polyhydroxybutyrate (PHB) nanoparticles modulate metals toxicity in Hydra viridissima.

The use of bioplastics (e.g., polyhydroxybutyrate) emerged as a solution to help reduce plastic pollution caused by conventional plastics. Nevertheless, bioplastics share many characteristics with their conventional counterparts, such as degradation to nano-sized particles and the ability to sorb environmental pollutants, like metals. This study aimed to assess the potential impacts of the interaction of metals (cadmium - Cd, copper - Cu, and zinc - Zn) with polyhydroxybutyrate nanoplastics (PHB-NPLs; ~200 nm) on the freshwater cnidarian Hydra viridissima in terms of mortality rates, morphological alterations, and feeding behavior. The metal concentrations selected for the combined exposures corresponded to concentrations causing 20 %, 50 %, and 80 % of mortality (LC20, LC50, and LC80, respectively) and the PHB-NPLs concentrations ranged from 0.01 to 1000 µg/L. H. viridissima sensitivity to the metals, based on the LC50's, can be ordered as: Zn < Cd < Cu. Combined exposure to metals and PHB-NPLs yielded distinct outcomes concerning mortality, morphological changes, and feeding behavior, uncovering metal- and dose-specific responses. The interaction between Cd-LCx and PHB-NPLs progressed from no effect at LC20,96h to an ameliorative effect at Cd-LC50,96h. Cu-LCx revealed potential mitigation effects (LC20,96h and LC50,96h) but at Cu-LC80,96h the response shifts to a potentiating effect. For Zn-LCx, response patterns across the combinations with PHB-NPLs were like those induced by the metal alone. PHB-NPLs emerged as a key factor capable of modulating the toxicity of metals. This study highlights the context-dependent interactions between metals and PHB-NPLs in freshwater environments while supporting the need for further investigation of the underlying mechanisms and ecological consequences in forthcoming research.

Short- and long-term toxicity of nano-sized polyhydroxybutyrate to the freshwater cnidarian Hydra viridissima.

The accumulation of increasingly smaller plastic particles in aquatic ecosystems is a prominent environmental issue and is causing a significant impact on aquatic biota. In response to this challenge, biodegradable plastics have emerged as a potential ecological alternative. Nevertheless, despite recent progress in polymer toxicology, there is still limited understanding of the ecological implications of biodegradable plastics in freshwater ecosystems. This study evaluated the toxicity of polyhydroxybutyrate nano-sized particles (PHB-NPLs) on the freshwater cnidarian Hydra viridissima assessing individual and population-level effects. Data revealed low toxicity of PHB-NPLs to H. viridissima in the short-term, as evidenced by the absence of significant malformations and mortality after the 96-h assays. In addition, hydras exhibited rapid and complete regeneration after 96 h of exposure to PHB-NPLs. Feeding assays revealed no significant alterations in prey consumption behavior in the 96-h mortality and malformations assay and the regeneration assay. However, significantly increased feeding rates were observed after long-term exposure, across all tested concentrations of PHB-NPLs. This increase may be attributed to the organisms' heightened energetic demand, stemming from prolonged activation of detoxification mechanisms. These changes may have a cascading effect within the food web, influencing community dynamics and ecosystem stability. Furthermore, a dose-dependent response on the hydras' populational growth was found, with an estimated 20 % effect concentration (EC20,8d) on this endpoint of 10.9 mg PHB-NPLs/L that suggests potential long-term impacts on the population's reproductive output and potential depression and local extinction upon long-term exposure to PHB-NPLs on H. viridissima. The obtained data emphasizes the importance of evaluating sublethal effects and supports the adoption of long-term assays when assessing the toxicity of novel polymers, providing crucial data for informed regulation to safeguard freshwater ecosystems. Future research should aim to unravel the underlying mechanisms behind these sublethal effects, as well as the impact of the generated degradation products.

Effects of petroleum-based and biopolymer-based nanoplastics on aquatic organisms: A case study with mechanically degraded pristine polymers.

Mismanaged plastic litter submitted to environmental conditions may breakdown into smaller fragments, eventually reaching nano-scale particles (nanoplastics, NPLs). In this study, pristine beads of four different types of polymers, three oil-based (polypropylene, PP; polystyrene, PS; and low-density polyethylene, LDPE) and one bio-based (polylactic acid, PLA) were mechanically broken down to obtain more environmentally realistic NPLs and its toxicity to two freshwater secondary consumers was assessed. Thus, effects on the cnidarian Hydra viridissima (mortality, morphology, regeneration ability, and feeding behavior) and the fish Danio rerio (mortality, morphological alterations, and swimming behavior) were tested at NPLs concentrations in the 0.001 to 100 mg/L range. Mortality and several morphological alterations were observed on hydras exposed to 10 and 100 mg/L PP and 100 mg/L LDPE, whilst regeneration capacity was overall accelerated. The locomotory activity of D. rerio larvae was affected by NPLs (decreased swimming time, distance or turning frequency) at environmentally realistic concentrations (as low as 0.001 mg/L). Overall, petroleum- and bio-based NPLs elicited pernicious effects on tested model organisms, especially PP, LDPE and PLA. Data allowed the estimation of NPLs effective concentrations and showed that biopolymers may also induce relevant toxic effects.

Bioplastics: known effects and potential consequences to marine and estuarine ecosystem services.

Bioplastics have been suggested as more sustainable alternatives to conventional, petroleum-based plastics. In this work, the available studies comparing effects of biopolymers and petroleum-based plastics were reviewed to improve the knowledge on the sustainability of biobased polymers, providing a benchmark regarding their ecotoxicological effects, as well as to highlight research priorities in this field. The literature review shows that, only a small number of the available biopolymers have been tested highlighting the need for more research diversifying the tested polymers. Overall, the available studies support the idea that bioplastics are likely to cause physiological impairments (feeding, reproduction, or locomotion) as well as cellular (proteome and enzyme activity) effects on biota. Furthermore, the studies on bioplastic degradation under realistic conditions report changes in water and sediment quality, which may also have consequences to biota. It is evident that some reservations must be kept regarding conventional plastics substitutions by bioplastics.

Ecotoxicity of cationic cellulose polymers to aquatic biota: The influence of charge density.

Better performances of cellulose-based polymers can be achieved by adjust their architecture including the density of cationic modifications. In this study, the influence of cationic substitution on the ecotoxicity of four quaternized hydroxyethyl cellulose polymers (SK-H, SK-L, SK-M, SK-MH) was studied, using an aquatic biota acute ecotoxicity classification, and rheological and physicochemical characterization. The ecotoxicity characterization was achieved by performing standard ecotoxicity assays with seven key trophic level species: Vibrio fischeri, Raphidocelis subcapitata, Chlorella vulgaris, Daphnia magna, Brachionus calyciflorus, Heterocypris incongruens, and Danio rerio. Median effective concentrations were used to compute hazard concentrations, through the species sensitive distribution curves method. The microalga C. vulgaris and rotifer B. calyciflorus were the most sensitive species to the studied polymers. The SK-H variant was highly toxic to the rotifer. Overall, variants with intermediate levels of cationic charge (SK-M, SK-MH) presented the lowest toxicity. The SK-M variant showed the lowest value of maximum acceptable concentration (0.00354 mg/L), thus being indicated as the least toxic variant. Therefore, the obtained results suggest that industry could direct the development of this type of polymers by tailoring its cationic substitution to moderate levels, in such a way that both functionality and environmental toxicity could be maximized.

Effects of virgin and weathered polystyrene and polypropylene microplastics on Raphidocelis subcapitata and embryos of Danio rerio under environmental concentrations.

Microplastics are ubiquitous contaminants of freshwater ecosystems. However, few ecotoxicity assays have been conducted on freshwater organisms using environmentally relevant concentrations of virgin and weathered microplastics. This work assessed the adverse effects of virgin and artificially weathered fragments of polystyrene and polypropylene on the microalga Raphidocelis subcapitata (72 h growth inhibition assay) and on embryos of the fish Danio rerio (96 h fish embryo assay) under environmentally relevant concentrations (2000-200,000 MP L-1) and high concentrations (12.5-100 mg L-1). Sizes of microplastics were measured as tens (polystyrene) to hundreds (polypropylene) of micrometers, while aging was assessed by measuring the carbonyl index. In the microalga, the tested high concentrations promoted growth, while environmentally relevant concentration induced either growth inhibition or promotion. In zebrafish embryos, environmentally relevant concentrations decreased body length and heart rates. No relevant effects were observed in organisms exposed to high concentrations for mortality, malformations, hatching rates, and swimming bladder inflation. Virgin microplastics presented slightly higher toxicity but direct comparison was hindered by the lack of a linear dose-response curve. Despite the lack of a clear pattern, adverse effects were often observed in the lowest environmentally relevant concentrations, raising concerns over the impacts of microplastics on freshwater ecosystems.

Public views on plastic pollution: Knowledge, perceived impacts, and pro-environmental behaviours.

Nowadays, plastic pollution is considered an issue of global concern. This environmental problem results from human industrial and domestic activities, associated with poor management, from manufacturing processes to products' end life. In this perspective, human consumption and management of daily used plastic materials can play a determinant role to control this environmental issue. Thus, understanding public perceptions about plastic pollution may be a valuable resource to engage society in solutions to reduce its environmental release. In this study, perceptions about plastic pollution, its impacts as well as sociodemographic and psychological factors predicting individuals' pro-environmental behaviours were analysed, in the Portuguese context. Overall, results showed that there is a general knowledge that plastics degrade in the environment. The participants of the study perceived the bio-ecological impacts of plastics as a greater threat than the socioeconomic impacts. A hierarchical regression analyses revealed that sociodemographic variables and those related to knowledge and perceived impacts about plastic pollution can predict participants' pro-environmental behaviours. Awareness about the impacts of plastic pollution (socioeconomic, health impacts and bio-ecological impacts) were highly associated to pro-environmental behaviour. This study data can help to understand how to enhance pro-environmental behaviours and contribute to decrease the presence of micro(nano)plastics in the environment.

On the path to minimize plastic pollution: The perceived importance of education and knowledge dissemination strategies.

Environmental awareness and education have been recognized as tools to help mitigate environmental issues. This study, performed in Portugal, aimed to describe perceived factors building environmental awareness on plastic pollution, study how perceptions are influenced by sociodemographic characteristics, and identify perceived priority targets for intervention. Results showed that participants recognize society's important role to raise awareness and to develop mitigation strategies. Higher perception was reported by women and participants with a higher level of school education. The more educated participants consider individuals possessing expert knowledge as a more reliable information source, whereas older and less schooled participants trust more in media messaging. Participants acknowledge that environmental education should be included in school plans, particularly in the first nine years of education (students with ages between 6 and 14, approximately). Data support the idea that target specific dissemination strategies should be planned according to variables like age, gender and education level.

Considerations when using microplates and Neubauer counting chamber in ecotoxicity tests on microplastics.

The ubiquity of microplastics raises issues regarding contamination control measures and laboratory practices. The objective was to adapt the use of counting chambers and plastic microplates on the ecotoxicity evaluation of microplastics. Counting chambers, originally used to quantify cells, can also be used to count high concentrations of microplastics (<100 µm) used in laboratory assays. By decontaminating the chamber and mixing the test solution with Nile Red (1:1), fluorescent particles can be easily counted under optical microscopy. Microplate wells, due to their composition, can be contaminated or release microplastics to the test medium, which can interfere with the results of ecotoxicity assays or spectroscopy readings. A cleaning method based on ethanol was developed, which effectively removed particles by 91% without interfering with microalgae yield. Besides providing practical applications that can improve ecotoxicity assays, this work intends to raise awareness on the need to adapt laboratory practices when working with microplastics.

Nanoplastics and marine organisms: What has been studied?

Nowadays, there is an increased awareness on the threat that marine litter may pose to the marine environment. This review describes the major concerns related to plastic pollution, namely in terms of toxicity of different types and sizes of nanoplastics (particles smaller than 100 nm) to marine organisms, either producers or consumers. The available data show that nanoplastics may affect negatively organisms from different phyla with reported effects ranging from alterations in reproduction to lethality. Nevertheless, no information regarding marine vertebrates (e.g., fish) was found. Data show a high potential for bioaccumulation/biomagnification along marine food chains, since they can easily be retained inside organisms. The lack of standardized methodology for nanoplastics detection and the poor or inexistent legislation makes nanoplastics an environmental challenge.