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Global environmental contamination by microplastics (MPs) is a growing problem with potential One Health impacts. The presence of MPs in vital organs, such as the heart, is of particular concern, but the knowledge is still limited. The goal of the present pilot study was to investigate the potential presence of MPs in the heart of wild specimens of three commercial fish species (Merluccius merluccius, Sardina pilchardus, and Trisopterus luscus) from the North East Atlantic Ocean. Heart samples from 154 fish were analysed for MP content (one heart sample per fish). A total of 44 MPs were recovered from heart samples from the three species. MPs had varied chemical composition (5 polymers), shapes (4) and colours (5). Differences in the profile of the MPs among species was observed (p ≤ 0.05). Thirty fish (19%) had MPs in their hearts, with a total mean (±SD) concentration of 0.286 ± 0.644 MPs/fish. S. pilchardus had the highest heart contamination (p ≤ 0.05). There were no significant (p > 0.05) differences between M. merluccius and T. luscus. These findings in fish with different biological and ecological traits together with literature data suggest that heart contamination likely is a disseminated phenomenon. Therefore, further research on the presence of MPs in the cardiovascular system and its potential health effects is very much needed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2023. Published by Elsevier Ltd.)

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 (BCF Fish >1), but their bioaccumulation did not occur (BAF Fish <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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

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., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

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., Competing Interests: Declaration of competing interest None., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

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 EC 50 s 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., Competing Interests: Declaration of competing interest None., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

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 EC 50 s 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'., Competing Interests: Declaration of competing interest None., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

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., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

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 EC 50 s 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., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

In the present study, the levels of bisphenol A (BPA) and analagous compounds in muscle and liver of fish (Dicentrarchus labrax, Trachurus trachurus, Scomber colias) from the North East Atlantic Ocean were determined and the risk of their consumption by humans was assessed. The potential relationship between bisphenol concentrations and microplastic (MP) contamination of fish was also investigated. Fish from all the species had BPA in the liver and muscle, and bisphenol B (BPB) and bisphenol E (BPE) in the muscle. The highest concentration of BPA in the liver (302 ng/g dry weight - dw) was found in S. colias and the lowest one (5 ng/g dw) in T. trachurus. In the muscle, the bisphenol with the highest concentration was BPE in S. colias (272 ng/g dw). Fish with microplastics had significantly higher concentrations of bisphenols than fish where no microplastics were found, suggesting a relation between MP and bisphenol contamination in fish. In all species, the concentration of bisphenols was correlated with higher MP intake. Regarding human food safety, the estimated daily intake (EDI), target hazard quotient (THQ) and hazard index (HI) of bisphenols were higher than those established by the European Food Safety Authority suggesting hazardous risk for human consumers. These findings highlight the need of more research on fish contamination by MP and associated chemicals and inherent human food safety risks., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

The widespread use of microplastics and nanomaterials resulting in environmental contamination is of high concern. Microplastics have been found to modulate the toxicity of other environmental contaminants. Thus, the hypothesis that microplastics increase the toxicity of gold nanoparticles to the marine microalgae Tetraselmis chuii was tested. In a laboratory bioassay, T. chuii cultures were exposed for 96 h to ∼5 nm diameter gold nanoparticles (AuNP) and to virgin 1-5 μm diameter microplastics (MP), alone and in mixture. The treatments were: control; citrate-control; AuNP alone (0.1, 0.3 and 3 mg/L); MP alone (0.3, 0.9 and 4 mg/L) and mixture of the two substances in three different concentrations (0.1 mg/L AuNP + 0.3 mg/L MP; 0.3 mg/L AuNP + 0.9 mg/L MP; 3 mg/l AuNP + 4 mg/L MP). The effect criterion was the inhibition of the average specific growth rate. AuNP alone and MP alone did not cause significant decrease of T. chui average specific growth rate up to 3 mg/L and 4 mg/L, respectively. The mixture containing 3 mg/L AuNP + 4 mg/L MP significantly reduced the average specific growth rate of the microalgae. Therefore, this mixture was more toxic to T. chuii than its components individually. Overall, the results of the present study indicated that the MP and AuNP tested have a relatively low toxicity to T. chuii, but the toxicity increases when they are in mixtures containing high concentrations of both substances. These proof-of-concept findings stress the need of more research on the toxicity of mixtures containing microplastics and nanomaterials., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Microplastics (MPs) were found to modulate the toxicity of other pollutants but the knowledge on the topic is still limited. The goals of this study were to investigate the short-term toxicity of cadmium (Cd) to wild Pomatochistus microps juveniles, the potential modulation of acute Cd toxicity by 1-5 µm polyethylene MPs in this species, and possible differences of sensitivity to Cd and MPs-Cd mixtures between juveniles from two distinct wild populations. Juveniles were collected in the estuaries of Minho (M-est) and Lima (L-est) Rivers (NW Portugal). One 96 h bioassay with M-est juveniles and another one with L-est juveniles were carried out in laboratory conditions. Each bioassay had 12 treatments: control, 5 Cd concentrations, 1 MPs concentration, and 5 MPs-Cd mixtures. No significant differences in Cd-induced mortality between juveniles from distinct estuaries or between juveniles exposed to Cd alone and those exposed to MPs-Cd mixtures were found. The total 96h LC 10 and LC 50 of Cd alone were 2 mg/L (95% CI: 0-4 mg/L) and 8 mg/L (95% CI: 2-17 mg/L), respectively. Cd alone significantly decreased the post-exposure predatory performance (PEPP) of M-est (≥6 mg/L) and L-est juveniles (≥3 mg/L), and acetylcholinesterase (AChE) activity of M-est juveniles (13 mg/L). MPs alone (0.14 mg/L) significantly reduced the PEPP and AChE activity of L-est juveniles but not of M-est juveniles. MPs-Cd mixtures (3-13 mg/L of Cd + 0.14 mg/L of MPs) significantly inhibited the PEPP of juveniles from both estuaries and AChE of L-est estuary juveniles but not of M-est juveniles. Evidences of toxicological interactions, namely antagonism, between MPs and Cd were found. Overall, the results indicate that MPs modulated the sub-lethal toxic effects of Cd in wild P. microps juveniles, especially neurotoxicity. Moreover, the environmental conditions of the natural habitats to which juveniles were exposed during pre-developmental phases influence the sub-lethal toxicity of Cd, MPs, and their mixtures. The implications to environmental and human risk assessment are discussed and further research is needed.

The main objectives of this study were to investigate the effects of a mixture of microplastics and mercury on Corbicula fluminea, the post-exposure recovery, and the potential of microplastics to influence the bioconcentration of mercury by this species. Bivalves were collected in the field and acclimated to laboratory conditions for 14 days. Then, a 14-day bioassay was carried out. Bivalves were exposed for 8 days to clean medium (control), microplastics (0.13 mg/L), mercury (30 µg/L) and to a mixture (same concentrations) of both substances. The post-exposure recovery was investigated through 6 additional days in clean medium. After 8 and 14 days, the following endpoints were analysed: the post-exposure filtration rate (FR); the activity of cholinesterase enzymes (ChE), NADP-dependent isocitrate dehydrogenase (IDH), octopine dehydrogenase, catalase, glutathione reductase, glutathione peroxidase and glutathione S-transferases (GST), and the levels of lipid peroxidation (LPO). After 8 days of exposure to mercury, the bioconcentration factors (BCF) were 55 in bivalves exposed to the metal alone and 25 in bivalves exposed to the mixture. Thus, microplastics reduced the bioconcentration of mercury by C. fluminea. Bivalves exposed to microplastics, mercury or to the mixture had significantly (p ≤ 0.05) decreased FR and increased LPO levels, indicating fitness reduction and lipid oxidative damage. In addition, bivalves exposed to microplastics alone had significant (p ≤ 0.05) reduction of adductor muscle ChE activity, indicating neurotoxicity. Moreover, bivalves exposed to mercury alone had significantly (p ≤ 0.05) inhibited IDH activity, suggesting alterations in cellular energy production. Antagonism between microplastics and mercury in FR, ChE activity, GST activity and LPO levels was found. Six days of post-exposure recovery in clean medium was not enough to totally reverse the toxic effects induced by the substances nor to eliminate completely the mercury from the bivalve's body. These findings have implications to animal, ecosystem and human health., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)

The environmental contamination by microplastics is a global challenge to ecosystem and human health, and the knowledge on the long-term effects of such particles is limited. Thus, the effects of microplastics and post-exposure recovery were investigated over 4 generations (F 0 , F 1 , F 2 , F 3 ) using Daphnia magna as model. Effect criteria were parental mortality, growth, several reproductive parameters, and population growth rate. Microplastics exposure (0.1mg/l of pristine polymer microspheres 1-5μm diameter) caused parental mortality (10-100%), and significantly (p≤0.05) decreased growth, reproduction, and population growth rate leading to the extinction of the microplastics-exposed model population in the F 1 generation. Females descending from those exposed to microplastics in F 0 and exposed to clean medium presented some recovery but up to the F 3 generation they still had significantly (p≤0.05) reduced growth, reproduction, and population growth rate. Overall, these results indicate that D. magna recovery from chronic exposure to microplastics may take several generations, and that the continuous exposure over generations to microplastics may cause population extinction. These findings have implications to aquatic ecosystem functioning and services, and raise concern on the long-term animal and human exposure to microplastics through diverse routes., (Copyright © 2018. Published by Elsevier B.V.)

Recent studies have demonstrated the negative impacts of microplastics on wildlife. Therefore, the presence of microplastics in marine species for human consumption and the high intake of seafood (fish and shellfish) in some countries cause concern about the potential effects of microplastics on human health. In this brief review, the evidence of seafood contamination by microplastics is reviewed, and the potential consequences of the presence of microplastics in the marine environment for human food security, food safety and health are discussed. Furthermore, challenges and gaps in knowledge are identified. The knowledge on the adverse effects on human health due to the consumption of marine organisms containing microplastics is very limited, difficult to assess and still controversial. Thus, assessment of the risk posed to humans is challenging. Research is urgently needed, especially regarding the potential exposure and associated health risk to micro- and nano-sized plastics., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The effects of emerging environmental contaminants on human and environmental health is of high concern, especially those potentially induced by mixtures. The main goal of the present study was to assess the chronic effects of mixtures of citrate stabilized ≈5 nm gold nanoparticles (AuNP) and 1-5μm microplastics (MP) on Daphnia magna. A 21-day bioassay was carried out. The effect criteria were parental mortality, somatic growth and several reproductive parameters. AuNP induced parental mortality, reduced the total offspring and caused immobile juveniles and aborted eggs. MP induced parental mortality, delayed the first brood release, decreased the number of broods released, the total offspring, and caused immobile juveniles. All the mixtures caused higher toxicity than AuNP and MP alone. Based on parental mortality, evidences of antagonism between AuNP and MP were observed at low concentrations of both mixture components, whereas evidences of synergism at high concentrations were found. Chronic (21-day) exposure of D. magna to AuNPs, MP, and their mixtures can impair development, reproduction, ultimately leading to death., (Copyright © 2018. Published by Elsevier B.V.)

Microplastics and antimicrobials are widely spread environmental contaminants and more research on their toxicity is needed. The uptake and effects of the antimicrobial florfenicol, microplastics, and their mixtures on Corbicula fluminea were investigated. Bivalves were exposed for 96h to florfenicol (1.8 and 7.1mg/l), microplastics (0.2 and 0.7mg/l), or mixtures of the two substances. After 96h, all bivalves exposed to antimicrobial treatments had florfenicol in their body (e.g. 2±1μg/g). Microplastics were found in the gut, lumen of the digestive gland, connective tissue, hemolymphatic sinuses, and gills surface of animals. Florfenicol caused a significant inhibition of cholinesterase (ChE) activity (~32%). Animals exposed to 0.2mg/l of microplastics showed ChE activity inhibition (31%), and no other significant alterations. Mixtures caused feeding inhibition (57-83%), significant ChE inhibition (44-57%) and of isocitrate dehydrogenase activity, and increased anti-oxidant enzymes activity and lipid peroxidation levels. Overall, the results indicate that C. fluminea take up florfenicol and microplastics from the water and accumulated or at least retained it in their body for some time; both florfenicol (low ppm range) and microplastics (ppb range) were toxic to C. fluminea, with mixtures containing florfenicol and microplastics being more toxic. Thus, the risk of exposure and toxic effects of florfenicol to C. fluminea and other bivalves, and its predators increase in ecosystems contaminated with the antimicrobial and microplastics, as well as to humans consuming contaminated species from these ecosystems., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The goal of this study was to investigate the toxicity of cefalexin to Pomatoschistus microps juveniles in relation to the presence of microplastics in the water and temperature rise. After acclimatization, groups of wild juveniles were exposed for 96h to artificial salt water (control), microplastics alone (0.184mg/l), cefalexin alone (1.3-10mg/l) and in mixture with microplastics (cefalexin: 1.3-10mg/l; microplastics: 0.184mg/l) at 20 and 25°C. Effect criteria were mortality, post-exposure predatory performance (PEPP), acetylcholinesterase activity (AChE) and lipid peroxidation levels (LPO). At 20°C, concentrations of cefalexin alone≥5mg/l significantly reduced PEPP (up to 56%; 96h-EC 50 =8.4mg/l), indicating toxicity of the antibiotic to juveniles after short-term exposure to water concentrations in the low ppm range. At 20°C, fish exposed to microplastics alone did not have significant differences in any of the parameters tested relative to the control group but tended to have an inhibition of the PEPP (23%) and AChE (21%); at 25°C, microplastics alone caused mortality (33%) and PEPP inhibition (28%). Thus, microplastics are toxic to P. microps juveniles. At 20°C, under simultaneous exposure to cefalexin and microplastics, the PEPP was significantly reduced (at cefalexin concentrations≥1.25mg/l). Moreover, at 25°C, the toxicity curves of cefalexin (PEPP based), alone and in mixture with microplastics, were significantly different (p<0.05; 96h-EC 50 of 3.8 and 5.2mg/l, respectively), and the integrated data analysis indicated significant interactions between the two substances for all biomarkers. Thus, the presence of microplastics in the water influenced the toxicity of cefalexin. The rise of water temperature (from 20°C to 25°C), increased the microplastics-induced mortality (from 8 to 33%), and the inhibitory effects of cefalexin on the PEPP (up to 70%). Significant differences (p<0.05) between the toxicity curves of cefalexin alone at distinct temperatures were found, with a lower 96h-EC 50 at 25°C (3.8mg/l) than at 20°C (8.4mg/l). Moreover, at 25°C, increases of AChE activity (14%) and LPO (72%) in fish exposed to the mixture treatment containing the highest cefalexin concentration were found, and the integrated analysis of data indicated significant interactions between cefalexin and temperature for PEPP, and among all stressors for LPO. Thus, the temperature rise increased the toxicity of microplastics and of cefalexin, alone and in mixture with microplastics, to P. microps juveniles. These findings raise concern on the long-term exposure of wild populations to complex mixtures of pollutants, likely decreasing their fitness, and highlight the need of more research on the combined effects of widely used pharmaceuticals, microplastics and temperature increase on wild species to improve environmental and human risk assessments of chemicals and their safe use under a global warming scenario., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Microplastics (MP) are ubiquitous contaminants able to cause adverse effects on organisms. Three hypotheses were tested here: early Pomatoschistus microps juveniles can ingest MP; the presence of MP may reduce fish predatory performance and efficiency; developmental conditions may influence the preyselection capability of fish. Predatory bioassays were carried out with juveniles from two estuaries with differences in environmental conditions: Minho (M-est) and Lima (L-est) Rivers (NW Iberian coast). Polyethylene MP spheres (3 types) alone and in combination with Artemia nauplii were offered as prey.All the MP types were ingested, suggesting confusion with food. Under simultaneous exposure to MP and Artemia, L-est fish showed a significant reduction of the predatory performance (65%) and efficiency (upto 50%), while M-est fish did not, suggesting that developmental conditions may influence the preyselection capability of fish. The MP-induced reduction of food intake may decrease individual and population fitness.

Large amounts of herbicides are presently used in the industrialized nations worldwide, with an inexorable burden to the environment, especially to aquatic ecosystems. Primary producers such as microalgae are of especial concern because they are vital for the input of energy into the ecosystem and for the maintenance of oxygen in water on which most of other marine life forms depend on. The herbicide paraquat (PQ) is known to cause inhibition of photosynthesis and irreversible damage to photosynthetic organisms through generation of reactive oxygen species in a light-dependent manner. Previous studies have led to the development of a new formulation of PQ containing lysine acetylsalicylate (LAS) as an antidote, which was shown to prevent the mammalian toxicity of PQ, while maintaining the herbicidal effect. However, the safety of this formulation to primary producers in relation to commercially available PQ formulations has hitherto not been established. Therefore, the aim of this study was to evaluate the toxicity of the PQ+LAS formulation in comparison with the PQ, using Chlorella vulgaris as a test organism. Effect criterion was the inhibition of microalgal population growth. Following a 96 h exposure to increasing concentrations of PQ, C. vulgaris growth was almost completely inhibited, an effect that was significantly prevented by LAS at the proportion used in the formulation (PQ+LAS) 1:2 (mol/mol), while the highest protection was achieved at the proportion of 1:8. In conclusion, the present work demonstrated that the new formulation with PQ+LAS has a reduced toxicity to C. vulgaris when compared to Gramoxone(®)., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The aim of this study was to develop two behavioral tests (swimming velocity and avoidance behavior) specific for the common prawn, Palaemon serratus, and to investigate the effects of sublethal concentrations of fenitrothion on behavior and on several biomarkers. In a first bioassay, mortality was investigated in prawns exposed during 96 h to concentrations of fenitrothion ranging from 39 to 40000 ng L(-1). The effects on swimming velocity and biomarkers were determined in prawns exposed to the sub-lethal concentrations of fenitrothion (from 39 to 625 ng L(-1)). A specific newly bioassay was developed to assess the capability of prawns to avoid the toxicant (avoidance test). Finally, in order to determine the effects on biomarkers during the avoidance test, prawns were collected at different times of exposure (30, 60, 90 and 120 min). Results showed that prawns exposed to the sub-lethal concentrations of fenitrothion exhibited a significant inhibition of swimming velocity with a LOEC of 313 ng L(-1). A significant inhibition of both eye AChE (LOEC=78 ng L(-1)) and muscle ChE (LOEC=156 ng L(-1)) activities were also observed. Results from the avoidance test indicated that animals significantly avoided fenitrothion (78 ng L(-1)). However, this capability was not observed in prawns exposed to 156 ng L(-1) fenitrothion. Prawns exposed to fenitrothion showed alterations in enzymes involved in the production of energy (LDH and IDH) possibly in an attempt to cope with additional energetic demands. Impairment of locomotion and avoidance may lead to alterations at the population level. Thus, the present behavioral tests can be useful as ecologically relevant tools for ecological risk assessment., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The main purpose of the present study was to investigate the effects of deltamethrin on biomarkers and behavior of Palaemon serratus (common prawn), since this attempt to link different levels of biological organization will allow determining which biomarkers might be ecologically relevant and will be useful to complement the information about the effects of pesticides by using behavioral parameters. Therefore, parameters of liver antioxidant status, energy metabolism and neurotransmission were determined in different tissues of the common prawn and used to assess the effects at sub-individual level, whereas swimming velocity was used to assess the effects at the individual level. It was also investigated if the swimming velocity can be used as an endpoint in ecotoxicology bioassays and if it can be as sensitive as biomarker endpoints. Swimming velocity was significantly reduced in prawns exposed to deltamethrin, showing a lowest observed effect (LOEC) of 0.6 ng L(-1). Eye acetylcholinesterase (AChE) activity was significantly increased in prawns exposed to 0.6, 1.2 and 2.4 ng L(-1) deltamethrin, whereas muscle cholinesterase (ChE) activity was significantly increased in prawns exposed to 19 and 39 ng L(-1). On the other hand, lactate dehydrogenase (LDH) activity was significantly increased in muscle of prawns exposed to 0.6, 1.2, 2.4, 4.9 ng L(-1) deltamethrin, showing that organisms were requiring additional energy, but probably using it for detoxification processes rather than locomotion, since swimming velocity was inhibited. Glutathione S-transferase (GST) activity was significantly increased in the digestive gland of common prawn exposed to 19 and 39 ng L(-1) deltamethrin. Catalase (CAT) activity was significantly increased in digestive gland of prawn exposed to 19 ng L(-1) deltamethrin. However, CAT activity decreased in digestive gland of prawn exposed to 39 ng L(-1), suggesting an antioxidant defense system failure concomitant with high levels of lipid peroxidation. Thus, global results showed that decreased swimming velocity was not associated with cholinesterase inhibition. In fact, the impairment of swimming velocity may be due to allocation of energy for detoxification and antioxidant protection instead of swimming activity. The present study showed that swimming velocity could be used as an ecologically relevant tool and a sensitive endpoint to assess and complement the study of pesticide effects on marine organisms., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Effects induced on wild populations by recurrent environmental contamination may difficult the ecological risk assessment of punctual pollution events such as oil spills. Here, the issue was addressed by comparing the health status of Pomatoschistus microps populations from four NW Iberian estuaries, using an integrated chemical-biological monitoring. Despite high seasonal variability, the parameters measured discriminated estuaries with different contamination levels and associated biological effects with chemical and abiotic stress. The decreased health status of fish from polluted sites strengthens the need of considering pollution-induced background effects and seasonal variability when assessing impacts and risks of oil and other chemical spills.

The aim of this study was to assess the effects of the widely used anti-depressant fluoxetine on behaviour (locomotion), moulting, neuromuscular transmission, energy production and anti-oxidant defences' efficiency of the epibenthic crab Carcinus maenas. Crabs were individually exposed to fluoxetine concentrations for 7d. Effects on locomotion were assessed at the end of the exposure using an open field test adapted to C. maenas in the present study. Tissue samples were later collected to evaluate fluoxetine effects on physiological functions using the activity of key enzymes and other parameters as biomarkers, namely: N-acetyl-β-glucosaminidase (NAGase) in the epidermis (moulting) and the hepatopancreas; cholinesterases (ChE) in muscle (neuromuscular cholinergic transmission); NADP(+)-dependent isocitrate dehydrogenase (IDH) and lactate dehydrogenease (LDH) in muscle (energy production); glutathione S-transferases (GST) in hepatopancreas (biotransformation and oxidative stress system); glutathione reductase (GR), and glutathione peroxidade (GPx), total glutathione levels (TG) and lipid peroxidation levels in the hepatopancreas (anti-oxidant defences and oxidative damage). Because no information on C. maenas NAGase activity was previously available, its variation during the moult cycle was also investigated. The results showed that locomotion was significantly increased at fluoxetine concentrations equal or above 120 μg L⁻¹, with animals spending more time moving, walking longer distances than controls. Levels of NAGase activity were found to vary in relation to C. maenas moult cycle, but no alterations were observed after exposure to fluoxetine. Significant increases in the activity of ChE, GST and GR enzymes, and the levels of TG were found, with a lowest observed effect concentration (LOEC) of 120 μg L⁻¹. Effects on locomotion were significantly and positively correlated to those induced on ChE activity. The results raise concern when hypothesising conditions of chronic exposure in the wild., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The present study was conducted to assess three microevolutionary aspects of adaptation to pollution in Daphnia longispina populations historically exposed to an acid mine drainage from an abandoned pyrite mine: pollution mediated effects in acute tolerance to copper (Cu) and zinc (Zn); pollution-mediated effects on genetic variability of tolerant and physiological traits related to fitness (feeding rates); and fitness costs of tolerance measured as genetic trade-offs between tolerance and feeding rates under none and low levels of contamination. These objectives were addressed by comparing broad sense heritabilities and genetic correlations using up to 20 distinct clonal lineages randomly obtained from two populations: one located in a water reservoir contaminated by the acid mine drainage, and the other located in a nearby clean water reservoir. Results showed that only sensitive and resistant lineages to Cu were present in the reference and contaminated site, respectively. For Zn, however, both populations had a similar distribution pattern of sensitivities. Heritability values for tolerant and feeding traits across metal exposure levels was similar in both populations being in most cases greater than 50%. Fitness costs of tolerance were illustrated by lower feeding rates of the tolerant population compared to the reference one and negative genetic correlations between mean clonal feeding rates and median clonal survival time in control conditions (no added Cu or Zn). The results obtained thus support the view that tolerance to pollution is ecologically costly., ((c) 2009 SETAC.)

In November 2002, the tanker 'Prestige' released about 19,000 tonnes of a heavy fuel oil (no. 6) before sinking with about 58,000 tonnes of its cargo, 135 miles from Cabo Finisterra (Spain). A considerable part of the released fuel oil reached the Galician coast, causing a heavy black tide and an ecological disaster. Although the black tide did not reach the NW coast of Portugal, it is possible that some of the fuel oil or its components also arrived to this area directly through the sea water and/or indirectly through the food chain. Therefore, the aim of this study was to investigate possible changes in two widely used biomarkers, the activity of the enzymes cholinesterases (ChE) and glutathione S-transferases (GST), of three molluscs (Mytilus galloprovincialis, Nucella lapillus and Monodonta lineata) from wild populations of the NW Portuguese coast in relation to the 'Prestige' oil spill. Molluscs were collected seasonally before (autumn 2002) and after (winter 2002/2003), spring and summer 2003) the oil spill at several sites along the Portuguese NW coast. Enzymatic activities determined before the accident were compared with those determined at different times after the oil spill taking into consideration abiotic factors. Information from different parameters was integrated by Redundancy Analysis and Principal Response Curves (PRC). Results show that GST and ChE activities were influenced by abiotic factors. Despite this influence, the results of PRC analysis also suggest that some of the fuel oil reached the NW Portuguese coast changing the patterns of ChE and GST activities of local populations of rocky shore species. Furthermore, the present study highlights the need of long-term monitoring with wild populations to assess both historical and punctual effects of pollution in the marine environment.

The study presented here searched for the level of taxonomic resolution required to detect the effects of low-level chronic pollution on estuarine meiobenthic communities. Meiofauna from two sites, with special attention to harpacticoid copepods, was analysed at different taxonomic levels of aggregation using uni- and multivariate methods. Adaptation processes that could buffer biodiversity disruptions were also considered through the analysis of fitness-related and tolerance traits in the harpacticoid copepod Paronychocamptus nanus. Results showed that uni- and multivariate analyses could be inadequate when assessing subtle anthropogenic contamination. Instead, the assessment of inter-population differences in tolerance to the main source of stress rises as a required procedure if potential effects of this type of contamination are being investigated. Specifically, a 96 h acute toxicity test performed with populations from the affected site appears as a faster and reliable general tool to assess impacts of low-level chronic pollution in estuaries.

In aquatic environments, many factors determine the overall impact of a toxic event, constituting "exposure scenarios" that must be understood so that risk assessment strategies can be successfully applied. In this work, an experimental design was planned to investigate the influence of the organism age, duration of exposure and subsequent recovery period (after cessation of toxic exposure) on chlorpyrifos or carbendazim toxicity to Kiefferulus calligaster. The endpoints studied were cholinesterase (ChE) activity and glutathione S-transferase (GST) activity. The influence of age and exposure time and the existence of recovery depend on the chemical and the biomarkers tested, suggesting that no general rule can be applied when trying to assess overall toxicity of pesticides and that detailed knowledge has to be acquired on a case-to-case basis.

Mercury is one of the most hazardous metals that may contaminate estuarine ecosystems and induce toxic effects on wildlife organisms. It has been suggested that impairment of cholinesterase (ChE) activity may be involved in the resulting mercury toxicity. Following Palaemon serratus exposure to mercury chloride (HgCl2), no effect on ChE activity was observed whatever the concentration used (to 37.5 microM) or the time of exposure (to 7 days). By contrast, following 24 h exposure to dichlorvos, an organophosphate insecticide with a well-characterised anti-ChE action, decrease of ChE activity was observed until 30 to 40% basal activity, which seems to be the minimum activity required for prawn survival. In addition, HgCl2 does not affect dichlorvos toxicity and treatments with a mixture of both compounds can be interpreted as the sum of the two independent toxicities. Therefore, mercury and insecticide toxicities are independent and ChE activity from P. serratus eyes seems to be a reliable and sensitive biomarker for organophosphate insecticides even when organisms are simultaneously exposed to mercury.

This study was conducted to investigate the potential use of biomarkers in bioassays with chironomids to assess contamination by pesticides in temperate and tropical climates. Two species of midge were studied, the widespread Chironomus riparius and the tropical Kiefferulus calligaster (Kieffer, 1911). Preliminary studies included investigations of the effects of temperature on larval development and the influence of larval age on normal variability of cholinesterase (ChE) and glutathione S-transferase (GST) activities and protein content. In the second phase, the influence of two abiotic factors particularly important in tropical conditions (temperature and oxygen concentration) and of the organophosphorous (OP) insecticide dimethoate on biochemical and conventional endpoints was investigated. Results showed that K. calligaster is morphologically and physiologically similar to C. riparius and for both, the time of larval development decreases with the increase of temperature. Moreover, 3rd and 4th instars appeared to be the most suitable for biomarkers determinations. ChE activity seems to be valuable biomarker regarding temperature and dissolved oxygen (DO) variations, while some caution should be taken when using GST as an environmental biomarker, since it shows some dependence of these parameters. C. riparius was more sensitive to dimethoate than K. calligaster suggesting that the use of bioassays with the former species in tropical conditions may overestimate the toxicity of OP pesticides to autochthonous species. When testing sub-lethal effects of dimethoate to C. riparius, ChE activity showed to be a very sensitive parameter detecting significant effects at the lowest concentration that caused emergence delay of larvae, suggesting that it is an ecologically relevant parameter.

Aquaculture systems are a potentially significant source of antibacterial agents to the aquatic environment. The antibacterials oxytetracycline (OTC) and florfenicol (FLO) have been widely used in aquaculture. These pharmaceuticals may cause deleterious effects on wild aquatic organisms accidentally exposed to them. Therefore, the objective of this study was to evaluate the acute toxicity of OTC and FLO to the microalgae Tetraselmis chuii and to the crustacean Artemia parthenogenetica, using culture growth inhibition and death, respectively, as effect criteria. OTC and FLO were found to inhibit the growth of T. chuii cultures, with 96 h IC(50) values of 11.18 and 6.06 mg/L, respectively. OTC 24 and 48 h LC(50) values for A. parthenogenetica were 871 and 806 mg/L, respectively. FLO did not cause mortality of A. parthenogenetica. These results indicate that OTC and FLO are considerably more toxic to T. chui than to A. parthenogenetica. They also indicate that the concentrations required to induce mortality to A. parthenogenetica only in exceptional conditions will occur in the environment.

Following the development of urban and industrial centres petrochemical products have become a widespread class of contaminants. The aim of this study was to investigate the effects of petrochemical contamination in wild populations of mussels (Mytilus galloprovincialis) along the NW Atlantic coast of Portugal by applying antioxidant and energetic metabolism parameters as biomarkers. For that, mussels were collected at five sampling sites presenting different petrochemical contamination levels. To evaluate the mussels' antioxidant status, enzymatic activities of catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, glutathione S-transferases, as well as glutathione redox status were evaluated in gills and digestive glands of mussels collected from the selected sites. Lipid peroxidation was determined in the same tissues to quantify cellular oxidative damage. Furthermore, to investigate how energetic processes may respond to these contaminants, the activity of NADP(+)-dependent isocitrate dehydrogenase was determined in mussels' digestive glands, and octopine dehydrogenase was determined in mussels' posterior adductor muscle. Furthermore, the concentrations of aliphatic hydrocarbons, unresolved complex mixture and polycyclic aromatic hydrocarbons (PAHs) were quantified in mussels' tissue, and abiotic parameters were quantified in water samples collected at each site. Several biomarkers showed statistically significant differences among sampling sites. The redundancy analysis (RDA) used to perform the integrated analysis of the data showed a clear separation of the sampling sites in three different assemblages, which are in agreement with the PAHs levels found in mussels tissues. In addition, the RDA indicated that some of the selected biomarkers may be influenced by abiotic parameters (e.g. salinity, pH, nitrates and ammonia). The approach selected for this study seems to be suitable for monitoring petrochemical contamination.

Information related to varied uses of several species of the genus Artemia (commonly known as brine shrimp), is dispersed among literature from several scientific areas, such as Ecology, Physiology, Ecotoxicology, Aquaculture and Genetics. The present paper reviews information related to Artemia that may be considered relevant for ecotoxicity testing. Integration of different areas of scientific knowledge concerning biology, life cycle and environmental needs of Artemia is of crucial importance when considering the interpretation of results drawn from tests involving this genus. Furthermore, this paper provides suggestions to overcome problems related to toxicity assessment with the use of Artemia as test organism in bioassays, under the scope of estuarine, marine and hypersaline environments. Aspects related to variability in results, adoptable toxicity end-points, culture conditions, characteristics of species and strains, influence of geographical origins over physiological features and responses to exposure to chemical agents are considered.

Given the urgent need for ecologically relevant toxicity tools specific to estuarine systems, the main goal of the present study was to develop and evaluate the potential of an in situ algal assay for estuarine sediment-overlying water toxicity evaluations using the microalga Phaeodactylum tricornutum immobilized in alginate beads. With its simple and cost-effective design, the assay apparatus, coupled with the use of microalgae immobilized in alginate beads, allowed rapid and easy deployment as well as retrieval of assay chambers. The effectiveness of the proposed in situ assay was investigated by deploying it at two reference and six contaminated Portuguese estuaries. By deploying open chambers both with and without an artificial nutrient source and closed chambers with an artificial seawater medium, it was possible to discriminate toxic effects from those caused by differences in temperature, light, and nutrient conditions across sites. A reduction in algal growth was found at contaminated sites relative to reference sites. The strontium alginate beads used for microalgae immobilization were found to be appropriate to perform in situ assays, showing no signs of disruption or dissolution after up to 8 d of exposure in estuarine sediment-overlying waters. The proposed in situ assay was shown to be a useful tool for estuarine sediment-overlying water toxicity testing to provide relevant data concerning primary production to estuarine ecological risk assessment studies.

This study aimed at integrating postexposure feeding and some biochemical parameters in the responses of the estuarine polychaeta, Hediste diversicolor, to controlled laboratory exposure conditions and to in situ exposures scenario of sediment contamination. Since H. diversicolor feeding may be considered as a major rate-limiting step in the processing of detritus in European estuaries, a reduction in feeding activity may have implications not only at the individual and population level of the species but also in detritus processing and in organic matter decomposition rates at the ecosystem level. The biochemical parameters were chosen as indicators of four key physiological functions: neurotransmission, metabolic condition, detoxification processes and antioxidant defences. The Mira and Sado estuaries, located in the Southwest coast of Portugal and classified as undisturbed and impacted, respectively, were selected as sites for this study. A significant depression in H. diversicolor postexposure feeding (from 30 to 70%) was consistently detected in all impacted sediments, supporting the sensitivity and responsiveness of feeding as a sublethal toxicity endpoint. Alongside with a reduced energy intake, an increased rate of organisms' anaerobic metabolism, as evidenced by an enhancement of lactate dehydrogenase activity (up to 1.5-fold), suggested a rapid need of additional energy to ameliorate chemical stress. Moreover, oxidative stress was shown to be an important mechanism of toxicity of the impacted sediments in H. diversicolor, as evidenced by a marked reduction in the glutathione redox status (up to 6.5-fold) and an increase in lipid peroxides levels (up to 2.3-fold) in organisms exposed to the most impacted sediments. Results of the in situ assay, conducted to assess the ecological relevance of sediment laboratory toxicity estimates and their application to make valid field extrapolations, revealed a lack of agreement in the response of catalase in organisms exposed to moderate impacted sediments. Our results support the utility of integrating responses at individual and sub-individual level to evaluate potential toxicant-induced changes in key physiological functions of H. diversicolor and to interpret their potential ecological consequences.

This study developed and evaluated a short-term sublethal in situ toxicity assay for estuarine sediment-overlying waters, with the crab Carcinus maenas (L.) based on postexposure feeding. It consisted of a 48-h in situ exposure period followed by a short postexposure feeding period (30 min). A precise method for quantifying feeding, using the Polychaeta Hediste (Nereis) diversicolor Müller as food source, was first developed. The sensitivity of the postexposure feeding response was verified by comparing it to that of lethality, upon cadmium exposure. The influence of environmental conditions prevailing during exposure (salinity, temperature, substrate, light regime, and food availability) on postexposure feeding was also addressed. The potential of this in situ assay was then investigated by deploying organisms at ten sites, located in reference and contaminated Portuguese estuaries. Organism recovery ranged between 90% and 100% and a significant postexposure feeding depression (16.3-72.7%) was observed at all contaminated sites relatively to references.

This study evaluated a short-term sublethal endpoint for in situ toxicity assays for estuarine sediments, based on postexposure feeding of the polychaete Hediste (Nereis) diversicolor Müller. A method for precisely quantifying postexposure feeding rates of H. diversicolor was established under laboratory conditions using Artemia franciscana Kellog nauplii. The sensitivity of the postexposure feeding response to copper was investigated by comparing postexposure feeding rates to growth and lethality. The 48-h and 96-h median lethal concentration (LC50) of copper were 241 and 125 microg/L, respectively, whereas the 48-h median inhibitory concentration (IC50) for postexposure feeding and the 20-d IC50 for growth were 52 and 25 microg/L of copper, respectively. The influence of different exposure conditions (substrate, temperature, salinity, food availability, and light) on H. diversicolor postexposure feeding was assessed; temperature and salinity were found to influence significantly postexposure feeding. The effectiveness of the proposed in situ assay was investigated by deploying it at two reference and six contaminated Portuguese estuaries. A 48-h exposure period was followed by a 1-h postexposure feeding period. High organism recoveries (89-100%) were obtained. Postexposure feeding was depressed significantly (17-90%) at all contaminated sites relatively to reference sites. The proposed in situ assay with H. diversicolor was shown to be a potential useful tool for estuarine sediment toxicity testing.

The interest in the ecological relevance of risk assessments and, thus, in in situ bioassays has been increasing in the last years. The present study developed a time- and cost-effective in situ bioassay, aiming at obtaining, in a short period of time and with a minimum of resources, a set of ecologically relevant toxicological information in a site-specific approach. Poecilia reticulata and Gambusia holbrooki were chosen as test species. Post-exposure feeding inhibition and the biomarkers acetylcholinesterase, lactate dehydrogenase and glutathione S-transferases were the endpoints tested. The battery of biomarkers as a whole was sensitive to the in situ exposure in an acid mine drainage impacted effluent, although responses varied between test species. Post-exposure feeding inhibition was the most sensitive endpoint, and its association with biomarker responses was discussed. The linkage between individual responses, such as feeding, and biomarkers suggested that, at least in this case, biomarkers can be relevant at higher levels of biological organization. Altogether, the proposed short-term in situ bioassay seems to be a promising tool, since it represents a reasonable compromise between sensitivity, time/cost-effectiveness and ecological relevance.

The toxicity of cholinesterase-inhibiting compounds (e.g., carbamates and organophosphates) is due to a decrease in acetylcholine metabolism, which results in a continuous stimulation of cholinergic receptors (muscarinic and nicotinic) that can be fatal. The goal of this study was to evaluate the protective effect of atropine (muscarinic receptor antagonist) against paraoxon-induced toxicity to Daphnia magna using its survival rate for the detection of environmental contamination by cholinesterase-inhibiting compounds. As expected, paraoxon was lethal to D. magna in a concentration-dependent manner. Noteworthy, the pretreatment of these organisms with atropine dramatically increased their survival against paraoxon. These results indicate that muscarinic stimulation plays an important role in paraoxon-induced lethality in D. magna. Therefore, simply by using the survival of atropine-treated and nontreated D. magna, water contamination by cholinesterase-inhibiting compounds may be rapidly and specifically detected., (Copyright 2002 Elsevier Science (USA))

The purpose of this study was to develop an in situ sediment bioassay chamber and respective procedures, suitable for performing toxicity bioassays with benthic invertebrates, using the midge Chironomus riparius. It was also our objective to compare the responses obtained under controlled conditions (laboratory 10-day larval growth and survival test) with those obtained in situ. Clean sand and a formulated sediment were incorporated in the in situ bioassay, along with local sediments, as a way of minimizing natural variability due to physicochemical differences among sediments or due to interactions with indigenous organisms. Recovery of organisms was good (80-100% in the control and reference site), indicating that the developed chamber and protocol were suitable for exposing and retrieving C. riparius in situ. Results also showed differences between responses obtained with formulated and natural sediment in situ, as well as between laboratory and in situ.