Microplastic contamination in marine mussels from the Atlantic coast of North Portugal and human risk of microplastic intake through mussel consumption.

Microplastic (MP) pollution has become a global concern due to its potential impacts on the environment, ecosystem services and human health. The goals of the present study were to document the MP contamination in wild specimens of Mytilus galloprovincialis sampled along the Atlantic coast of the North region of Portugal continental (NW Portuguese coast), and to estimate the human risk of MP intake (HRI) through the consumption of local mussels as seafood. Mussels were collected at four sampling sites along the NW Portuguese coast (40 mussels per site), and the whole soft body of each mussel was analysed for MP content. HRI estimates were based on the mean of MP items per wet weight of mussel analysed tissue (MP/g) and consumption habits. A total of 132 MP items were recovered from mussels. MP had diverse sizes (98-2690 µm) and colours. The most common shapes were fibres (39%) and pellets (36%). Five polymers were identified in the MP: polyethylene (50%), polystyrene (15%), poly(ethylene vinyl acetate) (14%), polyamide (12%) and polypropylene (9%). From the 160 analysed mussels, 55% had MP. The mean and standard error of the mean of mussel contamination ranged from 0.206 ± 0.067 and 0.709 ± 0.095 MP/g. Compared to estimates based on MP contamination in mussels from other areas and varied consumption habits, the HRI through the consumption of mussels from the NW Portuguese coast is relatively low.

Occurrence of microplastics and metals in European seabass produced in different aquaculture systems: Implications for human exposure, risk, and food safety.

Microplastics (MPs) are emerging contaminants of increasing concern as they may cause adverse effects and carry other contaminants, which may potentially compromise human health. Despite occurring in aquatic ecosystems worldwide, the knowledge about MP presence in different aquaculture systems and their potential impact on seafood products is still limited. This study aimed to determine the levels of MPs in water, feed, and European seabass (Dicentrarchus labrax) from three relevant aquaculture systems and estimate human exposure to MPs and metals through seabass consumption. The recirculating aquaculture system (RAS) had the highest MP occurrence in water and feed. MP levels in seabass followed the aquaculture system's levels in water and feed, with RAS-farmed fish presenting the highest MP load, both in the fish gastrointestinal tract (GIT) and muscle, followed by pond-, and cage-farmed fish. MPs' characteristics across aquaculture systems and fish samples remained consistent, with the predominant recovered particles falling within the MP size range. The particles were visually characterized and chemically identified by micro-Fourier Transform Infrared Spectroscopy (µFTIR). Most of these particles were fibres composed of man-made cellulose and PET. MP levels in GIT were significantly higher than in muscle for pond- and RAS-farmed fish, MPs' bioconcentration factors >1 indicated bioconcentration in farmed seabass. Metal concentrations in fish muscle were below permissible limits, posing low intake risks for consumers according to the available health-based guidance values and estimated dietary scenarios.

Microplastics in marine mussels, biological effects and human risk of intake: A case study in a multi-stressor environment.

This study documented seasonal levels of microplastics (MPs) and biomarkers (condition index, neurotoxicity, energy, oxidative stress) in mussels (Mytilus galloprovincialis), and water physico-chemical parameters in the Douro estuary (NE Atlantic coast), and estimated the human risk of MP intake (HRI) through mussels. Mussel stress was determined through the Integrated Biomarker Response (IBR). HRI was estimated from mussel MP concentrations and consumer habits. MPs were mainly micro-fibres (72 %) with varied chemical composition. Seasonal MP means (±SEM) in mussels ranged from 0.111 ± 0.044 (spring) to 0.312 ± 0.092 MPs/g (summer). Seasonal variations of mussel stress (IBR: 1.4 spring to 9.7 summer) and MP concentrations were not related. MeO-BDEs, PBDEs, temperature, salinity and other factors likely contributed to mussel stress variation. HRI ranged from 2438 to 2650 MPs/year. Compared to the literature, MP contamination in mussels is low, as well as the human risk of MP intake through their consumption.

Microplastics in water, feed and tissues of European seabass reared in a recirculation aquaculture system (RAS).

Plastic particles (PLs) are ubiquitous in aquatic ecosystems, and aquaculture production is susceptible to contamination from external or endogenous sources. This study investigated PL presence in water, fish feed and body sites of 55 European seabass produced in a recirculating aquaculture system (RAS). Fish morphometric parameters and health status biomarkers were determined. A total of 372 PLs were recovered from water (37.2 PL/L), 118 PLs from feed (3.9 PL/g), and 422 from seabass (0.7 PL/g fish; all body sites analysed). All 55 specimens had PLs in at least two of the four body sites analysed. Concentrations were higher in the gastrointestinal tract (GIT; 1.0 PL/g) and gills (0.8 PL/g) than in the liver (0.8 PL/g) and muscle (0.4 PL/g). PL concentration in GIT was significantly higher than in muscle. Black, blue, and transparent fibres made of man-made cellulose/rayon and polyethylene terephthalate were the most common PLs in water and seabass, while black fragments of phenoxy resin were the most common in feed. The levels of polymers linked to RAS components (polyethylene, polypropylene, and polyvinyl chloride) were low, suggesting a limited contribution to the overall PL levels found in water and/or fish. The mean PL size recovered from GIT (930 µm) and gills (1047 µm) was significantly larger than those found in the liver (647 µm) and dorsal muscle (425 µm). Considering all body sites, PLs bioconcentrated in seabass (BCFFish >1), but their bioaccumulation did not occur (BAFFish <1). No significant differences were observed in oxidative stress biomarkers between fish with low (<7) and high (≥7) PL numbers. These findings suggest that fish produced in RAS are mainly exposed to MPs through water and feed. Further monitoring under commercial conditions and risk assessment are warranted to identify potential threats to fish and human health and define mitigating measures.

Immunotoxicological effects of perfluorooctanesulfonic acid on European seabass are reduced by polyethylene microplastics.

Marine environments receive plastic waste, where it suffers a transformation process into smaller particles. Among them, microplastics (MPs; <5 mm) are ingested by aquatic organisms leading to negative effects on animal welfare. The interactions between MPs, contaminants and organisms are poorly understood. To clarify this issue, European seabass (Dicentrarchus labrax L.) were fed with diets supplemented with 0 (control), polyethylene (PE) MPs (100 mg/kg diet), perfluorooctanesulfonic acid (PFOS, 4.83 µg/kg diet) or PFOS adsorbed to MPs (MPs-PFOS; final concentrations of 4.83 µg and 100 mg of PFOS and MP per kg of feed, respectively). Samples of skin mucus, serum, head-kidney (HK), liver, muscle, brain and intestine were obtained. PFOS levels were high in the liver of fish fed with the PFOS-diet, and markedly reduced when adsorbed to MPs. Compared to the control groups, liver EROD activity did not show any significant changes, whereas brain and muscle cholinesterase activities were decreased in all the groups. The histological and morphometrical study on liver and intestine showed significant alterations in fish fed with the experimental diets. At functional level, all the experimental diets affected the humoral (peroxidase, IgM, protease and bactericidal activities) as well as cellular (phagocytosis, respiratory burst and peroxidase) activities of HK leukocytes, being more marked those effects caused by the PFOS diet. Besides, treatments produced inflammation and oxidative stress as evidenced at gene level. Principal component analysis demonstrated that seabass fed with MPs-PFOS showed more similar effects to MPs alone than to PFOS. Overall, seabass fed with MPs-PFOS diet showed similar or lower toxicological alterations than those fed with MPs or PFOS alone demonstrating the lack of additive effects or even protection against PFOS toxicity.

A global synthesis of microplastic contamination in wild fish species: Challenges for conservation, implications for sustainability of wild fish stocks and future directions.

Research on the occurrence of microplastics in wild fish populations is a constantly growing area, requiring continuous reviews to properly keep up with the fast pace of publications and guide future work. This review analyses the scientific output of 260 field studies covering 1053 different fish taxa for the presence of microplastics. To date, microplastics have been recorded in 830 wild fish species, including 606 species of interest to commercial and subsistence fisheries. Among these, based on IUCN Red List status, 34 species are globally classified in one of the three threatened categories (Critically Endangered, Endangered or Vulnerable) and another 22 species were assessed as "Near Threatened". Of the species for which the IUCN Red List tracks population trend data, the fish species reported to have microplastics so far include 81 which are recorded as declining, 134 as stable and just 16 as increasing. This review highlights the potential implications of fish microplastic contamination to biodiversity conservation, sustainability of wild fish stocks, and human food safety and security. Finally, recommendations for future research are presented.

Fate and effects of graphene oxide alone and with sorbed benzo(a)pyrene in mussels Mytilus galloprovincialis.

Graphene oxide (GO) has gained a great scientific and economic interest due to its unique properties. As incorporation of GO in consumer products is rising, it is expected that GO will end up in oceans. Due to its high surface to volume ratio, GO can adsorb persistent organic pollutants (POPs), such as benzo(a)pyrene (BaP), and act as carrier of POPs, increasing their bioavailability to marine organisms. Thus, uptake and effects of GO in marine biota represent a major concern. This work aimed to assess the potential hazards of GO, alone or with sorbed BaP (GO+BaP), and BaP alone in marine mussels after 7 days of exposure. GO was detected through Raman spectroscopy in the lumen of the digestive tract and in feces of mussels exposed to GO and GO+BaP while BaP was bioaccumulated in mussels exposed to GO+BaP, but especially in those exposed to BaP. Overall, GO acted as a carrier of BaP to mussels but GO appeared to protect mussels towards BaP accumulation. Some effects observed in mussels exposed to GO+BaP were due to BaP carried onto GO nanoplatelets. Enhanced toxicity of GO+BaP with respect to GO and/or BaP or to controls were identified for other biological responses, demonstrating the complexity of interactions between GO and BaP.

Are microplastics contributing to pollution-induced neurotoxicity? A pilot study with wild fish in a real scenario.

Pollution-induced neurotoxicity is of high concern. This pilot study investigated the potential relationship between the presence of microplastics (MPs) in the brain of 180 wild fish (Dicentrarchus labrax, Platichthys flesus, Mugil cephalus) from a contaminated estuary and the activity of the acetylcholinesterase (AChE) enzyme. MPs were found in 9 samples (5% of the total), all of them from D. labrax collected in the summer, which represents 45% of the samples of this species collected in that season (20). Seventeen MPs were recovered from brain samples, with sizes ranging from 8 to 96 µm. Polyacrylamide, polyacrylic acid and one biopolymer (zein) were identified by Micro-Raman spectroscopy. Fish with MPs showed lower (p ≤ 0.05) AChE activity than those where MPs were not found. These findings point to the contribution of MPs to the neurotoxicity induced by long-term exposure to pollution, stressing the need of further studies on the topic to increase 'One Health' protection.

Warmer water, high light intensity, lithium and microplastics: Dangerous environmental combinations to zooplankton and Global Health?

Nowadays there is a high concern about the combined effects of global warming and emerging environmental contaminants with significant increasing trends of use, such as lithium (Li) and microplastics (MPs), both on wildlife and human health. Therefore, the effects of high light intensity (26,000 lx) or warmer water temperature (25 °C) on the long-term toxicity of Li and mixtures of Li and MPs (Li-MPs mixtures) were investigated using model populations of the freshwater zooplankton species Daphnia magna. Three 21-day bioassays were done in the laboratory at the following water temperatures and light intensities: (i) 20 °C/10830 lx; (ii) 20 °C/26000 lx (high light intensity); (iii) 25 °C/10830 lx (warmer temperature). Based on the 21-day EC50s on reproduction, high light intensity increased the reproductive toxicity of Li and Li-MPs mixtures by ~1.3 fold; warmer temperature increased the toxicity of Li by ~1.2 fold, and the toxicity of Li-MPs mixtures by ~1.4 fold based on the concentration of Li, and by ~2 fold based on the concentrations of MPs. At high light intensity, Li (0.04 mg/L) and Li-MPs mixtures (0.04 Li + 0.09 MPs mg/L) reduced the population fitness by 32 % and 41 %, respectively. Warmer temperature, Li (0.05 mg/L) and Li-MPs mixtures (0.05 Li + 0.09 MPs mg/L) reduced it by 63 % and 71 %, respectively. At warmer temperature or high light intensity, higher concentrations of Li and Li-MPs mixtures lead to population extinction. Based on the population growth rate and using data of bioassays with MPs alone done simultaneously, Li and MPs interactions were antagonistic or synergistic depending on the scenario. High light intensity and chemical stress generally acted synergistically. Warmer temperature and chemical stress always acted synergistically. These findings highlight the threats of long-term exposure to Li and Li-MPs mixtures to freshwater zooplankton and Global Health in a warmer world.

Occurrence and risk assessment of endocrine-disrupting compounds in fish muscle: The case study of the Douro River estuary (North East Atlantic Ocean).

This work was focused on evaluating the occurrence of endocrine-disrupting compounds (EDCs) in fish muscles, such as bisphenol analogues, personal care products, including some UV-filters and musk fragrances, and selected pesticides. A total of 238 muscle samples of three fish species (Platichthys flesus, Mugil cephalus, and Dicentrarchus labrax) of an important estuary ending in the North East Atlantic Ocean (Douro River estuary, Portugal), in different seasons of the year, was analysed. M. cephalus was the species with a greater occurrence of contaminants, mainly in fishes collected during the spring. Bisphenol A was the main bisphenol analogue detected with an average content of 23.1 µg/kg wet weight (w.w.). Among chemicals used in personal care products, galaxolide, tonalide, and 2-ethylhexyl salicylate, were the most often found, being present in 15.9% (2.2-538.4 µg/kg w. w.), 4.6% (1.1-57.8 µg/kg w. w.) and 3.4% (3.9-56.1 µg/kg w. w.) of the samples, respectively. About 14% of the samples contained residues of at least one pesticide, being alachlor, aldrin, p,p'-DDT, permethrin, and prochloraz the main chemicals observed, varying from 0.1 µg/kg w. w. (p,p'-DDT) to 37.8 µg/kg w. w. (prochloraz). The daily intake estimates of the most frequent EDCs found in the fish muscles suggested that there are no health concerns, based on the recommended weekly consumption of fish for adults.

Effects of microplastics alone or with sorbed oil compounds from the water accommodated fraction of a North Sea crude oil on marine mussels (Mytilus galloprovincialis).

Microplastics (MPs) can adsorb persistent organic pollutants such as oil hydrocarbons and may facilitate their transfer to organisms (Trojan horse effect). The aim of this study was to examine the effects of a 21 day dietary exposure to polystyrene MPs of 4.5 µm at 1000 particles/mL, alone and with sorbed oil compounds from the water accommodated fraction (WAF) of a naphthenic North Sea crude oil at two dilutions (25 % and 100 %), on marine mussels. An additional group of mussels was exposed to 25 % WAF for comparison. PAHs were accumulated in mussels exposed to WAF but not in those exposed to MPs with sorbed oil compounds from WAF (MPs-WAF), partly due to the low concentration of PAHs in the studied crude oil. Exposure to MPs or to WAF alone altered the activity of enzymes involved in aerobic (isocitrate dehydrogenase) and biotransformation metabolism (glutathione S-transferase). Prevalence of oocyte atresia and volume density of basophilic cells were higher and absorption efficiency lower in mussels exposed to MPs and to WAF than in controls. After 21 days MPs caused DNA damage (Comet assay) in mussel hemocytes. In conclusion, a Trojan horse effect was not observed but both MPs and oil WAF caused an array of deleterious effects on marine mussels at different levels of biological organization.

Occurrence and seasonal variation of several endocrine disruptor compounds (pesticides, bisphenols, musks and UV-filters) in water and sediments from the estuaries of Tagus and Douro Rivers (NE Atlantic Ocean coast).

Exposure of aquatic environments to emerging contaminants is a global issue, special relevant in many estuaries due to impacts from anthropogenic activity. The aim of this work was to evaluate thirty-seven endocrine disruptor chemicals (EDCs) from four different classes (pesticides, bisphenols, polycyclic musks and UV-filters) in water and sediment samples collected during one-year in the estuaries of Tagus and Douro Rivers located into the NE Atlantic Ocean coast. EDCs analysis was achieved afterward validation of a gas-chromatography mass spectrometry (GC-MS) method using Dispersive Liquid-Liquid Microextraction (DLLME) as extraction procedure for water samples, and Quick, Easy, Cheap, Efficient, Rugged and Safe (QuEChERS) combined with DLLME for sediments. Tagus estuary presented higher levels of contamination with pesticide residues and bisphenols (BPs) than the Douro estuary in both water and sediment samples. Contrariwise, levels and frequency of polycyclic musks (PCMs) and UV-filters (UVF) were slightly higher in Douro estuary. Levels of pesticide residues in both sediment and water samples, and levels of PCMs and UVF in water samples were higher in warmer seasons (summer and spring) than in colder ones (winter and autumn). The opposite was found in what respect levels of BPs in water and sediment samples, and PCMs and UVF levels in sediment samples. Although the levels found for each contaminant are low, usually in the order of a few ng/mL(g), the presence of a high number of toxic compounds is a source of concern and requires constant monitoring.

Long-term effects of lithium and lithium-microplastic mixtures on the model species Daphnia magna: Toxicological interactions and implications to 'One Health'.

Environmental contamination with lithium (Li) and microplastics (MP) has been steadily increasing and this trend is expected to continue in the future. Many freshwater ecosystems, which are crucial to reach the United Nations Sustainable Development Goals, are particularly vulnerable to Li and MP contamination, and other pressures. The long-term effects of Li, either alone or combined with MP (Li-MP mixtures), were investigated using the freshwater zooplankton micro-crustacean Daphnia magna as model species. In the laboratory, D. magna females were exposed for 21 days to water concentrations of Li (0.02, 0.04, 0.08 mg/L) or Li-MP mixtures (0.02 Li + 0.04 MP, 0.04 Li + 0.09 MP mg/L, 0.08 Li + 0.19 MP mg/L). In the range of concentrations tested, Li and Li-MP mixtures caused parental mortality, and decreased the somatic growth (up to 20% and 40% reduction, respectively) and the reproductive success (up to 93% and 90% reduction, respectively). The 21-day EC50s of Li and Li-MP mixtures on D. magna reproduction were 0.039 mg/L and 0.039 Li + 0.086 MP mg/L, respectively. Under exposure to the highest concentration of Li (0.08 mg/L) and Li-MP mixtures (0.08 Li + 0.19 MP mg/L), the mean of D. magna population growth rate was reduced by 67% and 58%, respectively. Based on the population growth rate and using data from a bioassay testing the same concentrations of MP alone and carried simultaneously, the toxicological interaction between Li and MP was antagonism under exposure to the lowest and the highest concentrations of Li-MP mixtures, and synergism under exposure to the medium concentration of Li-MP mixtures. These findings highlight the need of further investigating the combined effects of contaminants, and the threat of long-term environmental contamination with Li and MP to freshwater zooplankton, biodiversity, ecosystem services and 'One Health'.

Microplastics in fishes from an estuary (Minho River) ending into the NE Atlantic Ocean.

Wild fish (Cyprinus carpio, Mugil cephalus, Platichthys flesus) from an estuary of the NE Atlantic coast were investigated for plastic contamination (N = 128). From the 1289 particles recovered from fish samples, 883 were plastics. Among these, 84% were fibres and 97% were microplastics. Thirty-six polymers were identified. The number of microplastics (mean ± SD) per individual fish (MP/fish) was 8 ± 6 in C. carpio, 10 ± 9 in M. cephalus and 2 ± 2 in P. flesus. The means of MP/fish per body site were 6 ± 7 in gastrointestinal tract, 0.5 ± 1.1 in gills, 0.3 ± 0.7 in liver and 0.6 ± 1.2 in muscle samples. A few large fibres in liver (≤ 4841 µm) and muscle (≤ 5810 µm) samples were found. The results evidence the existence of high fish contamination by microplastics and reinforce the need of further research on plastic pollution in estuaries.

Polybrominated diphenyl ethers and their methoxylated congeners in Douro river estuary biota: Seasonal occurrence and risk assessment.

Especially added on many industrial and domestic products as flame retardants (FRs), polybrominated diphenyl ethers (PBDEs) are among the chemicals of high environmental concern because of their potential harmfulness for environmental and human health. Seafood consumption is considered the main source of PBDEs and their methoxylated congeners (MeO-BDEs) for humans. The present study aims to investigate the seasonal occurrence of six PBDEs and eight MeO-BDEs congeners using Douro river biota (different trophic levels) as sentinels, as well as to evaluate the human exposure risk to PBDEs through seafood consumption. Biota samples (n = 273) were collected from one of the most important Portuguese estuaries in the north-western coast of Portugal at four different seasons (2019-2020). The analyses were performed by an environmental-friendly extraction procedure followed by Gas Chromatography coupled to a triple quadrupole detector (GC-MS/MS). PBDEs were detected in all seafood samples analysed, with means ranging from 0.02 ng g-1 ww (flounder in autumn) to 3.75 ng g-1 ww (mussel in winter). Levels of lower-brominated PBDE congeners were significantly higher than higher-brominated ones in all seasons (p < 0.01). MeO-BDEs ranged from 0.001 ng g-1 ww (grey mullet in summer) to 5.66 ng g-1 ww (green crab in spring). Crabs and mussels presented the highest means of PBDEs and MeO-BDEs. Regarding the health risk assessment of the studied PBDE congeners (47, 99, and 153), consumption of Douro river fish is not a case of concern for consumers.

Long-term adverse effects of microplastics on Daphnia magna reproduction and population growth rate at increased water temperature and light intensity: Combined effects of stressors and interactions.

In many ecosystems, the zooplankton community has been pressured simultaneously by microplastic pollution and alterations resulting from global climate changes. The potential influence of light intensity rise (from 10,830 lx to 26,000 lx) and water temperature rise (from 20 °C to 25 °C) on the long term-toxicity of microplastics (MPs) to Daphnia magna were investigated. Three 21-day laboratory bioassays with model MPs (1-5 µm diameter) were carried out at (i) 20 °C/10830 lx, (ii) 20 °C/26000 lx, and (iii) 25 °C/10830 lx. In each bioassay, one control (no MPs) and three MP concentrations (0.04, 0.09, 0.19 mg/L) were tested. In all the bioassays, MPs caused parental and juvenile mortality, and reduced the somatic growth, reproduction and population growth rate. The MP EC50s on living offspring (95% confidence interval within brackets) were 0.146 mg/L (0.142-0.151 mg/L) at 20 °C/10830 lx, 0.102 mg/L (0.099-0.105 mg/L) at 20 °C/26000 lx, and 0.101 mg/L (0.098-0.104 mg/L) at 25 °C/10830 lx. Relatively to the respective control group, 0.19 mg/L of MPs decreased the mean of the population growth rate by 27% at 20 °C/10830 lx, 38% at 20 °C/26000 lx and 59% at 25 °C/10830 lx. Based on the population growth rate and in relation to 20 °C/10830 lx (control, no MPs), the interaction between increased light intensity (26,000 lx) and MPs was synergism (at all the MP concentrations tested). The interaction between water temperature rise (25 °C) and MPs was antagonism at 0.04 mg/L of MPs and synergism at 0.09 and 0.19 mg/L of MPs. In the present scenario of climate changes and global MP pollution such findings raise high concern because zooplankton communities are crucial for aquatic biodiversity conservation, ecosystem functioning and services provided to humans. Further studies on the combined effects of MPs, other common pollutants, and alterations due to climate changes are needed.

Sorption of okadaic acid lipophilic toxin onto plastics in seawater.

The present study tested under laboratorial conditions the sorption of okadaic acid (OA), a lipophilic toxin produced by marine phytoplankton, onto 6-mm plastic circular fragments. Fragments (4 g L-1) of polyethylene terephthalate (PET), polypropylene (PP), expanded polystyrene (EPS) and non-expanded polystyrene (PS) were exposed for 96 h to seawater spiked with 10 ng mL-1 of OA. Results of the experiments showed a broad percentage of OA removed from the water by plastics after 48 h of exposure: 30 ± 5.1% (PET), 37 ± 9.5% (PP), 62 ± 7.1% (EPS) and 83 ± 1.9% (PS). Sorption appears to be highly influenced by polymer's characteristics, such as polarity and degree of crystallinity. Further studies are needed to clarify the effect of the contact area on sorption by expanded plastics. These results point to the plausible interaction of OA and plastics in coastal waters.