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Dr. Vera L. Maria (others: V. L. Maria or Maria)
Department of Biology & CESAM, University of Aveiro, Portugal

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0 Biomarkers
0 Ecotoxicology
0 Nanotoxicology
0 Zebrafish
0 OMICS

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Journal article
Published: 05 August 2021 in Applied Sciences
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Worldwide, pesticides have contaminated the environment, affecting non-target species. The aim of this work was to evaluate the effects of fosetyl-Al (FOS) on model organisms. Based on the 3 Rs for animal research and described guidelines, the OECD 236 and 220 were applied with some modifications. The FOS test concentrations were 0.02–0.2–2–20–200 mg/L for Danio rerio and 250–500–750–1000–1250 mg/kg for Enchytraeus crypticus. Besides the standard endpoints, additional endpoints were evaluated (D. rerio: behavior and biochemical responses; E. crypticus: extension of exposure duration (28 d (days) + 28 d) and organisms’ sizes). For D. rerio, after 96 h (h), hatching was inhibited (200 mg/L), proteins’ content increased (2 and 20 mg/L), lipids’ content decreased (2 mg/L), glutathione S-transferase activity increased (2 mg/L), and, after 120 h, larvae distance swam increased (20 mg/L). For E. crypticus, after 28 d, almost all the tested concentrations enlarged the organisms’ sizes and, after 56 d, 1250 mg/kg decreased the reproduction. In general, alterations in the organisms’ biochemical responses, behavior, and growth occurred at lower concentrations than the effects observed at the standard endpoints. This ecotoxicological assessment showed that FOS may not be considered safe for the tested species, only at higher concentrations than the predicted environmental concentrations (PECs). This research highlighted the importance of a multi-endpoint approach to assess the (eco)toxic effects of the contaminants.

ACS Style

Angela Barreto; Joana Santos; Mónica Amorim; Vera Maria. Is the Synthetic Fungicide Fosetyl-Al Safe for the Ecotoxicological Models Danio rerio and Enchytraeus crypticus? Applied Sciences 2021, 11, 7209 .

AMA Style

Angela Barreto, Joana Santos, Mónica Amorim, Vera Maria. Is the Synthetic Fungicide Fosetyl-Al Safe for the Ecotoxicological Models Danio rerio and Enchytraeus crypticus? Applied Sciences. 2021; 11 (16):7209.

Chicago/Turabian Style

Angela Barreto; Joana Santos; Mónica Amorim; Vera Maria. 2021. "Is the Synthetic Fungicide Fosetyl-Al Safe for the Ecotoxicological Models Danio rerio and Enchytraeus crypticus?" Applied Sciences 11, no. 16: 7209.

Journal article
Published: 31 July 2021 in Aquatic Toxicology
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Engineered nanoparticles (NPs) are emerging contaminants of concern and it is important to understand their environmental behavior and ecological risks to exposed organisms. Despite their ubiquitous presence in the environment, there is little information about the hazards of certain NPs, such as boron (BNPs) and vanadium (VNPs). The aim of the present research was to investigate the effects of commercial BNPs and VNPs (80 to 100 nm) to zebrafish embryos, at different levels of biological organization. A range of nominal concentrations for both NPs (0, 0.01, 0.1, 1, and 10 mg/L) was tested. Due to the presence of triton X-100 in the NPs’ stock dispersions, an additional control group was included (0.001% triton X-100). Survival, hatching, and malformations of embryos were assessed for 96 hours (h) exposure. Locomotor behavior was evaluated at 120 h. Furthermore, embryos were exposed to 0, 1, and 10 mg/L of NPs to evaluate a set of biomarker responses after 96 h: cholinesterase (ChE) and glutathione S-transferase (GST) activities, total glutathione (TG) and energy budgets levels. VNPs induced malformations (10 mg/L), hyperactivity (10 mg/L), erratic swimming (0.01 mg/L), altered swimming pattern (>0.01 mg/L), delayed hatching (10 mg/L) and altered biochemical responses involved in antioxidant defense (GST and TG at >1 mg/L), neurotransmission (ChE at 10 mg/L) and energy metabolism (lipids at >1 mg/L and carbohydrates at 10 mg/L). BNPs caused malformations (10 mg/L), affected swimming pattern (>0.01 mg/L), induced erratic swimming (10 mg/L) and decreased TG content and GST activity (>1 mg/L). At the same concentrations, VNPs affected a greater number of endpoints than BNPs, demonstrating a greater toxicity to zebrafish embryos. The present study shows that BNPs and VNPs may affect aquatic organisms, albeit at relatively great non-environmentally relevant concentrations, reinforcing the importance of the risk assessment of different NPs.

ACS Style

Joana Santos; Ângela Barreto; Célia Almeida; Cátia Azevedo; Inês Domingues; Mónica J.B. Amorim; Vera L. Maria. Toxicity of boron and vanadium nanoparticles on Danio rerio embryos – Phenotypical, biochemical, and behavioral alterations. Aquatic Toxicology 2021, 238, 105930 .

AMA Style

Joana Santos, Ângela Barreto, Célia Almeida, Cátia Azevedo, Inês Domingues, Mónica J.B. Amorim, Vera L. Maria. Toxicity of boron and vanadium nanoparticles on Danio rerio embryos – Phenotypical, biochemical, and behavioral alterations. Aquatic Toxicology. 2021; 238 ():105930.

Chicago/Turabian Style

Joana Santos; Ângela Barreto; Célia Almeida; Cátia Azevedo; Inês Domingues; Mónica J.B. Amorim; Vera L. Maria. 2021. "Toxicity of boron and vanadium nanoparticles on Danio rerio embryos – Phenotypical, biochemical, and behavioral alterations." Aquatic Toxicology 238, no. : 105930.

Journal article
Published: 28 July 2021 in Nanomaterials
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From the start of the 21st century, nanoecotoxicological research has been growing in fast steps due to the need to evaluate the safety of the increasing use of engineered nanomaterials. Boron (B) and vanadium (V) nanoparticles (NPs) generated by anthropogenic activities are subsequently released in the environment; therefore, organisms can be continuously exposed to these NPs for short or long periods. However, the short and long-term effects of BNPs and VNPs on soil organisms are unknown. This work aimed to recognize and describe their potential toxicological effects on the model species Enchytraeus crypticus, assessing survival and reproduction, through a longer-term exposure (56 days (d)—OECD test extension of 28 d), and avoidance behavior, through a short-term exposure (48 hours (h)). After 28 d, BNPs did not induce a significant effect on E. crypticus survival, whereas they decreased the organisms’ reproduction at 500 mg/kg. From 10 to 500 mg/kg, VNPs decreased the E. crypticus survival and/or reproduction. After 56 d, 100 to 500 mg/kg BNPs and 50 to 500 mg/kg VNPs, decreased the reproduction output of E. crypticus. The estimated Effect Concentrations (ECx) based on reproduction, for BNPs, were lower at 56 d compared with 28 d; for VNPs, an opposite pattern was found: ECx 28 d < ECx 56 d. BNPs did not induce an avoidance behavior, but organisms avoided the soil contaminated with 10 mg VNPs/kg. The tested NPs showed different E. crypticus apical effects at 28 d from the ones detected at 56 d, dependent on the type of NPs (B vs. V). In general, VNPs showed to be more toxic than BNPs. However, the effects of VNPs were alleviated during the time of exposure, contrarily to BNPs (which became more toxic with extended duration). The present study adds important information about NPs toxicity with ecological significance (at the population level). Including long-term effects, the obtained results contributes to the improvement of NPs risk assessment.

ACS Style

Angela Barreto; Joana Santos; Mónica Amorim; Vera Maria. Environmental Hazards of Boron and Vanadium Nanoparticles in the Terrestrial Ecosystem—A Case Study with Enchytraeus crypticus. Nanomaterials 2021, 11, 1937 .

AMA Style

Angela Barreto, Joana Santos, Mónica Amorim, Vera Maria. Environmental Hazards of Boron and Vanadium Nanoparticles in the Terrestrial Ecosystem—A Case Study with Enchytraeus crypticus. Nanomaterials. 2021; 11 (8):1937.

Chicago/Turabian Style

Angela Barreto; Joana Santos; Mónica Amorim; Vera Maria. 2021. "Environmental Hazards of Boron and Vanadium Nanoparticles in the Terrestrial Ecosystem—A Case Study with Enchytraeus crypticus." Nanomaterials 11, no. 8: 1937.

Journal article
Published: 15 June 2021 in Environmental Pollution
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Silver nanomaterials (AgNMs) are broadly used and among the most studied nanomaterials. The underlying molecular mechanisms (e.g. protein and metabolite response) that precede phenotypical effects have been assessed to a much lesser extent. In this paper, we assess differentially expressed proteins (DEPs) and metabolites (DEMs) by high-throughput (HTP) techniques (HPLC-MS/MS with tandem mass tags, reversed-phase (RP) and hydrophilic interaction liquid chromatography (HILIC) with mass spectrometric detection). In a time series (0, 7, 14 days), the standard soil model Enchytraeus crypticus was exposed to AgNM300K and AgNO3 at the reproduction EC20 and EC50. The impact on proteins/metabolites was clearly larger after 14 days. NM300K caused more upregulated DEPs/DEMs, more so at the EC20, whereas AgNO3 caused a dose response increase of DEPs/DEMs. Similar pathways were activated, although often via opposite regulation (up vs down) of DEPs, hence, dissimilar mechanisms underlie the apical observed impact. Affected pathways included e.g. energy and lipid metabolism and oxidative stress. Uniquely affected by AgNO3 was catalase, malate dehydrogenase and ATP-citrate synthase, and heat shock proteins (HSP70) and ferritin were affected by AgNM300K. The gene expression-based data in Adverse Outcome Pathway was confirmed and additional key events added, e.g. regulation of catalase and heat shock proteins were confirmed to be included. Finally, we observed (as we have seen before) that lower concentration of the NM caused higher biological impact. Data was deposited to ProteomeXchange, identifier PXD024444.

ACS Style

Vera L. Maria; David Licha; Janeck J. Scott-Fordsmand; Christian G. Huber; Mónica J.B. Amorim. Multiomics assessment in Enchytraeus crypticus exposed to Ag nanomaterials (Ag NM300K) and ions (AgNO3) – Metabolomics, proteomics (& transcriptomics). Environmental Pollution 2021, 286, 117571 .

AMA Style

Vera L. Maria, David Licha, Janeck J. Scott-Fordsmand, Christian G. Huber, Mónica J.B. Amorim. Multiomics assessment in Enchytraeus crypticus exposed to Ag nanomaterials (Ag NM300K) and ions (AgNO3) – Metabolomics, proteomics (& transcriptomics). Environmental Pollution. 2021; 286 ():117571.

Chicago/Turabian Style

Vera L. Maria; David Licha; Janeck J. Scott-Fordsmand; Christian G. Huber; Mónica J.B. Amorim. 2021. "Multiomics assessment in Enchytraeus crypticus exposed to Ag nanomaterials (Ag NM300K) and ions (AgNO3) – Metabolomics, proteomics (& transcriptomics)." Environmental Pollution 286, no. : 117571.

Journal article
Published: 26 February 2021 in Toxics
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Once in the environment, nanoplastics (NPls) may interact with other contaminants, such as pharmaceuticals, potentially acting as carriers and modulating their toxicity. Thus, the main aim of the current study is to investigate how polystyrene (PS) NPls (mean diameter: 60 nm) interact with simvastatin (SIM), an anticholesterolemic drug, and modulate its toxicity to zebrafish (Danio rerio) embryos. PS NPls were carboxyl group functionalized, to promote the interaction/binding of NPls with SIM (worst-case scenarios) and it was fluorescently dyed, allowing to detect the intake. Exposure was 96 h to 0–150 mg/L NPls or 0–150 µg/L SIM, as well as to dual combinations (NPls 0.015 or 1.5 mg/L and SIM 12.5 or 15 µg/L). PS NPls alone did not exert effects whereas SIM (≥ 12.5 µg/L) significantly delayed the hatching, decreased the heartbeat, induced edemas and mortality. The combination of NPls (1.5 mg/L) and SIM (12.5 or 15 µg/L) had significant effects on the survival of the organisms while the correspondent NPls and SIM single exposures did not have significant effects on this endpoint. Concerning the malformations appearance, SIM alone had similar effects than when in co-exposures (0.015 mg/L NPls plus 12.5 or 15 µg/L SIM). Hatching and heartbeat increased after the co-exposures SIM and NPls comparing with SIM single exposures, showing that 0.015 mg/L NPls plus 12.5 or 15 µg/L SIM did not cause significant effects on these endpoints. This study shows that NPls effects on bioavailability and toxicity of other contaminants cannot be ignored when assessing the environmental behavior and risks of NPls.

ACS Style

Angela Barreto; Joana Santos; Mónica Amorim; Vera Maria. Polystyrene Nanoplastics Can Alter the Toxicological Effects of Simvastatin on Danio rerio. Toxics 2021, 9, 44 .

AMA Style

Angela Barreto, Joana Santos, Mónica Amorim, Vera Maria. Polystyrene Nanoplastics Can Alter the Toxicological Effects of Simvastatin on Danio rerio. Toxics. 2021; 9 (3):44.

Chicago/Turabian Style

Angela Barreto; Joana Santos; Mónica Amorim; Vera Maria. 2021. "Polystyrene Nanoplastics Can Alter the Toxicological Effects of Simvastatin on Danio rerio." Toxics 9, no. 3: 44.

Journal article
Published: 30 October 2020 in Applied Sciences
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Nanoplastics (NPls) are ubiquitous in terrestrial environments, with numerous consequences for biodiversity and ecosystems. Research is urgently required to clarify the NPls environmental behaviour, fate and ecotoxicological effects to soil ecosystems. The aim of this research was to assess and comprehend the effects of polystyrene NPls to the terrestrial species Enchytraeus crypticus using survival, reproduction and avoidance behaviour as endpoints. A range of concentrations, 0.015 to 1500 mg NPls/kg LUFA 2.2 (Landwirtschaftliche Untersuchungs- und Forschungsanstalt Speyer, Germany) soil, was tested. Due to the effect of tween 20 and sodium azide (NaN3) on the NPls dispersion, the effects of these compounds were also assessed separately. After 21 d, 1200 and 1500 mg/kg NPls dispersion had significant effects on the organism survival and/or reproduction. However, these effects may be mainly associated with tween 20 and NaN3 present in the NPls dispersion and not with NPls themselves. After 48 h, there was a tendency of the organisms to avoid the NPls spiked soils, being this response significant at 0.015 mg/kg although a reduced avoidance behaviour was observed as NPls concentration increased. The present study provides screening data on the effects of NPls, alone and considering the presence of other compounds like the solvents, which is essential for regulators and strategic management of plastic pollution.

ACS Style

Angela Barreto; Joana Santos; Mónica Amorim; Vera Maria. How Can Nanoplastics Affect the Survival, Reproduction, and Behaviour of the Soil Model Enchytraeus crypticus? Applied Sciences 2020, 10, 7674 .

AMA Style

Angela Barreto, Joana Santos, Mónica Amorim, Vera Maria. How Can Nanoplastics Affect the Survival, Reproduction, and Behaviour of the Soil Model Enchytraeus crypticus? Applied Sciences. 2020; 10 (21):7674.

Chicago/Turabian Style

Angela Barreto; Joana Santos; Mónica Amorim; Vera Maria. 2020. "How Can Nanoplastics Affect the Survival, Reproduction, and Behaviour of the Soil Model Enchytraeus crypticus?" Applied Sciences 10, no. 21: 7674.

Journal article
Published: 25 February 2020 in Nanomaterials
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Silica nanoparticles (SiO2NPs) are one of the most used in commercial products and biomedical tools, however, their environmental effects have not been fully described. Although negative effects of SiO2NPs on the behavior of freshwater invertebrates have been reported, the knowledge is limited, especially the effect of nanopowders in terrestrial organisms. Accordingly, the aim of the present study is to understand the effects of SiO2NPs on the avoidance behavior of five soil species, whose niche may differ thus contributing to differential harmful SiO2NPs effects. Hence, avoidance assays testing SiO2NPs concentrations of 0, 10, 100, 250, 500 and 1000 mg/kg were performed with Enchytraeus crypticus, Folsomia candida, Tenebrio molitor, Porcellionides pruinosus and Eisenia fetida. SiO2NPs induced different behavioral effects, depending on the invertebrate ecology/habitat, exposure route and physiology. T. molitor, P. pruinosus and F. candida did not avoid contaminated soil; however, E. crypticus and E. fetida significantly avoided SiO2NPs spiked soil. Since these terrestrial worms (oligochaetes) live mostly burrowed in the soil, this can provide greater opportunity for SiO2NPs’ uptake. On the other hand, the other tested organisms mainly living on the upper part of the soil did not avoid the SiO2NPs spiked soil. The avoidance data obtained here also highlight the need for further studies to understand whether (or not) the detected behavioral responses are linked to either neurotransmission processes or sensorial aspects of the biological models.

ACS Style

Joana Santos; Ângela Barreto; João Nogueira; Ana Luísa Daniel-Da-Silva; Tito Trindade; Mónica J. B. Amorim; Vera L. Maria. Effects of Amorphous Silica Nanopowders on the Avoidance Behavior of Five Soil Species—A Screening Study. Nanomaterials 2020, 10, 402 .

AMA Style

Joana Santos, Ângela Barreto, João Nogueira, Ana Luísa Daniel-Da-Silva, Tito Trindade, Mónica J. B. Amorim, Vera L. Maria. Effects of Amorphous Silica Nanopowders on the Avoidance Behavior of Five Soil Species—A Screening Study. Nanomaterials. 2020; 10 (3):402.

Chicago/Turabian Style

Joana Santos; Ângela Barreto; João Nogueira; Ana Luísa Daniel-Da-Silva; Tito Trindade; Mónica J. B. Amorim; Vera L. Maria. 2020. "Effects of Amorphous Silica Nanopowders on the Avoidance Behavior of Five Soil Species—A Screening Study." Nanomaterials 10, no. 3: 402.

Journal article
Published: 23 October 2018 in Applied Soil Ecology
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The potential implications of long-term exposure to contaminants are not covered by current standard toxicity guidelines, usually referring to one generation and a fraction of the life cycle of the test species. Hence, in the present study, we aimed to assess the effects of the multigenerational exposure (generation 1–3: F1-F3) of Folsomia candida to an insect growth regulator (IGR) compound: teflubenzuron (TFB). The selected endpoints included both the standard ones as in the OECD and ISO guidelines (survival, reproduction, and avoidance) as well as additional ones (organisms size and cellular oxidative stress markers: acetylcholinesterase, glutathione S-transferase, catalase, glutathione peroxidase, lipid peroxidation). Although no avoidance behaviour was recorded at field-relevant concentrations (PEC (Predicted Environmental Concentration) = 0.06 mg/kg soil dry weigh (dwt)), survival and reproduction were impacted (LC50 = 0.1 mg/kg soil dwt; EC50 = 0.05 mg/kg soil dwt). Multigenerational exposure to TFB caused increased toxicity in F. candida in F3 in terms of survival and reproduction. This could be related to the mode of action of TFB which does not seem to activate some of the general stress mechanisms of response like oxidative stress. In addition, TFB causes a reduction of the organisms’ size, with a reduction of the number of large-sized juveniles, which has potential adverse consequences in terms of organisms’ performance, e.g. change in age structure and hence population dynamics. Hence, both observations may increase the environmental concern and associated risk of this insecticide.

ACS Style

Bruno Guimarães; Vera Lúcia Maria; J. Römbke; M.J.B. Amorim. Exposure of Folsomia candida (Willem 1902) to teflubenzuron over three generations – Increase of toxicity in the third generation. Applied Soil Ecology 2018, 134, 8 -14.

AMA Style

Bruno Guimarães, Vera Lúcia Maria, J. Römbke, M.J.B. Amorim. Exposure of Folsomia candida (Willem 1902) to teflubenzuron over three generations – Increase of toxicity in the third generation. Applied Soil Ecology. 2018; 134 ():8-14.

Chicago/Turabian Style

Bruno Guimarães; Vera Lúcia Maria; J. Römbke; M.J.B. Amorim. 2018. "Exposure of Folsomia candida (Willem 1902) to teflubenzuron over three generations – Increase of toxicity in the third generation." Applied Soil Ecology 134, no. : 8-14.

Journal article
Published: 25 September 2018 in Geoderma
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In standard toxicity tests one generation of test organisms is used, and they are usually exposed only during a fraction of their life-cycle. This approach is very important but does not cover the potential effects of multigenerational (MG) exposure and may underestimate risks. Hence, the main aim of this study was to assess the MG impact of the veterinary pharmaceutical ivermectin (IVM) on Folsomia candida during three generations (F1–F3). Ivermectin is a veterinary medicine, persistent in the environment and toxic to non-target soil invertebrates. A suite of different endpoints was used including avoidance, survival, reproduction, size and cellular biomarkers (catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST), acetylcholinesterase (AChE) and lipid peroxidation (LPO)). Survival and reproduction were affected (LC50: 40 mg/kg; EC50: 5 mg/kg), but no avoidance occurred, which poses additional ecological concern. Exposure throughout the generations showed similar toxicity in terms of survival and reproduction. Regarding size there was an impact, e.g., organisms were smaller and more abundant in F2 and larger and less abundant in F3. This can have implications in terms of risk as e.g. smaller organisms can respond differently to stress compared to larger organisms in future generations. The antioxidant mechanisms were dynamically activated along the generations, e.g. in F1 CAT was increased whereas in F3 there was increased GST activity, which resulted in damage (LPO) for F1 and F2 organisms but not for the F3 generation. The multi-endpoint approach proved to be beneficial for the interpretation of results and we recommend it, especially for persistent and/or highly adsorptive chemicals, but also endocrine disruptors. Moreover, the evaluation of size as an additional sub-lethal endpoint has significantly added to the relevance of this test. The relative proportion of small, medium and large animals may be an even more relevant aspect of this endpoint. This does not require guideline modifications and is hence easily implementable.

ACS Style

Bruno Guimarães; Vera Lúcia Maria; J. Römbke; M.J.B. Amorim. Multigenerational exposure of Folsomia candida to ivermectin – Using avoidance, survival, reproduction, size and cellular markers as endpoints. Geoderma 2018, 337, 273 -279.

AMA Style

Bruno Guimarães, Vera Lúcia Maria, J. Römbke, M.J.B. Amorim. Multigenerational exposure of Folsomia candida to ivermectin – Using avoidance, survival, reproduction, size and cellular markers as endpoints. Geoderma. 2018; 337 ():273-279.

Chicago/Turabian Style

Bruno Guimarães; Vera Lúcia Maria; J. Römbke; M.J.B. Amorim. 2018. "Multigenerational exposure of Folsomia candida to ivermectin – Using avoidance, survival, reproduction, size and cellular markers as endpoints." Geoderma 337, no. : 273-279.

Research article
Published: 07 September 2018 in Environmental Science & Technology
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Tungsten carbide cobalt (WCCo) nanoparticles (NPs) are widely used in hard metal industries. Pulmonary diseases and risk of cancer are associated with occupational exposure, but knowledge about the environmental fate and effects is virtually absent. In this study, the fate and effects of crystalline WCCo NPs, WC, and Co2+ were assessed in the soil model Enchytraeus crypticus, following the standard Enchytraeid Reproduction Test (ERT). An additional 28 day exposure period compared to the ERT (i.e., a total of 56 days) was performed to assess longer-term effects. WCCo NPs affected reproduction at a concentration higher than the corresponding Co based (EC50 = 1500 mg WCCo/kg, equivalent to 128 mg Co/kg). WC showed no negative effect up to 1000 mg W/kg. Maximum uptake of Co was 10-fold higher for CoCl2 compared to WCCo exposed organisms. Overall toxicity seems to be due to a combined effect between WC and Co. This is supported by the soil bioavailable fraction and biological tissue measurements. Last, results highlight the need to consider longer exposure period of NPs for comparable methods standardized for conventional chemicals.

ACS Style

Maria J. Ribeiro; Vera Lúcia Maria; Amadeu M.V.M. Soares; Janeck Scott-Fordsmand; Mónica J.B. Amorim. Fate and Effect of Nano Tungsten Carbide Cobalt (WCCo) in the Soil Environment: Observing a Nanoparticle Specific Toxicity in Enchytraeus crypticus. Environmental Science & Technology 2018, 52, 11394 -11401.

AMA Style

Maria J. Ribeiro, Vera Lúcia Maria, Amadeu M.V.M. Soares, Janeck Scott-Fordsmand, Mónica J.B. Amorim. Fate and Effect of Nano Tungsten Carbide Cobalt (WCCo) in the Soil Environment: Observing a Nanoparticle Specific Toxicity in Enchytraeus crypticus. Environmental Science & Technology. 2018; 52 (19):11394-11401.

Chicago/Turabian Style

Maria J. Ribeiro; Vera Lúcia Maria; Amadeu M.V.M. Soares; Janeck Scott-Fordsmand; Mónica J.B. Amorim. 2018. "Fate and Effect of Nano Tungsten Carbide Cobalt (WCCo) in the Soil Environment: Observing a Nanoparticle Specific Toxicity in Enchytraeus crypticus." Environmental Science & Technology 52, no. 19: 11394-11401.

Accelerated article
Published: 24 August 2018 in PROTEOMICS
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The transcriptome of the ecotoxicological model Enchytraeus crypticus (known as potworm) is well studied but the downstream changes at the protein level remained a gap. Changes in the protein regulation following exposure to CuO nanomaterial (NM) and Cu salt (CuCl2) were investigated. High performance liquid chromatography with tandem mass spectrometry using tandem mass tags was used. CuO NM elicited higher number of differentially expressed proteins compared to CuCl2 with little to no overlap of proteins. CuO NM caused more stress response mechanisms, with good agreement between differentially expressed proteins, genes and metabolites. CuCl2 caused higher impact in shorter time periods, but organisms have conserved mechanisms (constitutive genes) that allow Cu handling and detoxification. CuO NM caused higher impact after a longer exposure period, inducing regulation of facultative genes with a whole differentiated paradigm and cascade. This could be due to different issues: 1) the cell uptake route is different for Cu NM and Cu ions, 2) internalized Cu NM can result in a “Trojan‐ horse” effect, 3) the cascade of events occurs in a different time order, 4) the organism uptake is different between life stages, i.e., cocoons thickened surface protects the entry of NM and juveniles have facilitated entry via tegument. This article is protected by copyright. All rights reserved

ACS Style

Vera L. Maria; David Licha; Janeck Scott-Fordsmand; Christian G. Huber; Mónica J.B. Amorim. The Proteome ofEnchytraeus crypticus-Exposure to CuO Nanomaterial and CuCl2-in Pursue of a Mechanistic Interpretation. PROTEOMICS 2018, 18, e1800091 .

AMA Style

Vera L. Maria, David Licha, Janeck Scott-Fordsmand, Christian G. Huber, Mónica J.B. Amorim. The Proteome ofEnchytraeus crypticus-Exposure to CuO Nanomaterial and CuCl2-in Pursue of a Mechanistic Interpretation. PROTEOMICS. 2018; 18 (19):e1800091.

Chicago/Turabian Style

Vera L. Maria; David Licha; Janeck Scott-Fordsmand; Christian G. Huber; Mónica J.B. Amorim. 2018. "The Proteome ofEnchytraeus crypticus-Exposure to CuO Nanomaterial and CuCl2-in Pursue of a Mechanistic Interpretation." PROTEOMICS 18, no. 19: e1800091.

Journal article
Published: 01 August 2018 in Science of The Total Environment
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Effects of pollutants are mostly assessed using standard testing procedures, which cover a fraction of the animals' life cycle. Although, in nature species are exposed during multiple generations of sub-lethal doses of persistent chemicals. In the present study, we focused on the multigenerational (MG) effects of silver in Folsomia candida during 6 generations using the EC50 for reproduction as exposure concentration. We tested 9 different exposure scenarios, going from continuous 6 generations Ag exposure over pulse exposure (i.e. one generation clean, next contaminated, next clean etc.) to gradually increasing the number of exposure generations, with a final transfer to clean media. The biological endpoints assessed included survival, reproduction and size, with reproduction being the most sensitive. The biological response depended on the specific MG scenario, e.g. the 6 Ag MG caused a decreased number of juveniles from F4, whereas the pulse exposure experienced an increase in reproductive output when in clean soil. It is uncertain whether Ag causes transgenerational effects, but the reproduction levels in both pulse exposures are lower than in continuous control over the 6 generations which could be due to transference of Ag by the maternal generation. Overall, population size distribution seemed to indicate a delay in time for egg laying, with close relationship between adult survival, organisms size and reproduction output. Size monitoring allowed significant added interpretation possibilities and we strongly recommend the addition of this endpoint to the standard guideline. The smaller observed size range can have implications in terms of adaptation potential, carrying associated increased risk.

ACS Style

Luís André Mendes; Vera Lúcia Maria; Janeck Scott-Fordsmand; M.J.B. Amorim. Multigenerational exposure of Folsomia candida to silver: Effect of different contamination scenarios (continuous versus pulsed and recovery). Science of The Total Environment 2018, 631-632, 326 -333.

AMA Style

Luís André Mendes, Vera Lúcia Maria, Janeck Scott-Fordsmand, M.J.B. Amorim. Multigenerational exposure of Folsomia candida to silver: Effect of different contamination scenarios (continuous versus pulsed and recovery). Science of The Total Environment. 2018; 631-632 ():326-333.

Chicago/Turabian Style

Luís André Mendes; Vera Lúcia Maria; Janeck Scott-Fordsmand; M.J.B. Amorim. 2018. "Multigenerational exposure of Folsomia candida to silver: Effect of different contamination scenarios (continuous versus pulsed and recovery)." Science of The Total Environment 631-632, no. : 326-333.

Article
Published: 22 June 2018 in Nanotoxicology
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The stress metabolome provides a thorough insight into the signals and hence mechanisms of response of organisms. This is an excellent tool to advance the understanding of interactions, especially for substances like nanomaterials (NMs), for which there is an urgent need for alternative methods for hazard assessment. The metabolome of Enchytraeus crypticus was studied for the first time. The case study, CuO NM (and CuCl2) covered exposure along a time frame [0–7–14 days (d)] and two reproduction effect concentrations (EC10 and EC50). High-performance liquid chromatography-mass spectrometry based method (HPLC-MS) was used, with reversed phase (RP) separation and mass spectrometric detection in positive and negative modes. Metabolite profiling of Cu materials yielded 155 and 382 metabolite features in positive and negative modes, respectively, showing an expression related with time, material, and ECx. The number of differentially expressed metabolites (DEMs) decreased with exposure time (14 d) for CuO NM, whereas for CuCl2 EC50 it increased. Overall, almost all DEMs are down-regulated for CuO NM and up-regulated for CuCl2 (both modes). Early effects were mainly related to amino acids and later to lysophospholipids (down-regulation). Furthermore, the underlying mechanisms of CuO NM toxicity (e.g. neurotransmission, nucleic acids generation, cellular energy, and immune defense) differ from CuCl2, where later metabolomic responses are mostly linked to the metabolism of lipids and fewer to amino acids. This study reports a large scale metabolome profiling for E. crypticus and identifies potential markers of Cu materials, which can help to align intelligent testing strategies and safer-by-design materials.

ACS Style

Vera L. Maria; David Licha; Christina Ranninger; Janeck Scott-Fordsmand; Christian G. Huber; Mónica J. B. Amorim. The Enchytraeus crypticus stress metabolome – CuO NM case study. Nanotoxicology 2018, 12, 766 -780.

AMA Style

Vera L. Maria, David Licha, Christina Ranninger, Janeck Scott-Fordsmand, Christian G. Huber, Mónica J. B. Amorim. The Enchytraeus crypticus stress metabolome – CuO NM case study. Nanotoxicology. 2018; 12 (7):766-780.

Chicago/Turabian Style

Vera L. Maria; David Licha; Christina Ranninger; Janeck Scott-Fordsmand; Christian G. Huber; Mónica J. B. Amorim. 2018. "The Enchytraeus crypticus stress metabolome – CuO NM case study." Nanotoxicology 12, no. 7: 766-780.

Environmental toxicology
Published: 10 August 2017 in Environmental Toxicology and Chemistry
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Enchytraeids have been used in standard ecotoxicity testing for approximately 20 yr. Since adopting the standard test for survival and reproduction, a number of additional tools have been developed, including transcriptomics and enzymatic biomarkers. So far, a genotoxicity tool and endpoint have not been used; hence, the goals of the present study included optimization of the in vivo alkaline comet assay in Enchytraeus crypticus. Further, the effect of silver nanomaterial (Ag NM300K, dispersed, 15 nm) was tested and compared with silver nitrate. Hydrogen peroxide was used as a positive control. The various steps were optimized. The fully detailed standard operating procedure is presented. Silver materials caused genotoxicity, this being differentiated for the nano and non‐nano forms. Silver nitrate caused genotoxicity after 3 d of exposure in a dose‐related manner, although after 7 d the effects were either reduced or repaired. Ag NM300K caused higher genotoxicity after 7 d for the lowest concentration, highlighting a potential nonmonotonic dose–response effect. Overall, the comet assay showed the power to discriminate effects between materials and also toxicity at low relevant doses. Environ Toxicol Chem 2018;37:184–191. © 2017 SETAC

ACS Style

Vera L. Maria; Maria João Ribeiro; Sofia Guilherme; Amadeu M.V.M. Soares; Janeck Scott-Fordsmand; Mónica Amorim. Silver (nano)materials cause genotoxicity in Enchytraeus crypticus , as determined by the comet assay. Environmental Toxicology and Chemistry 2017, 37, 184 -191.

AMA Style

Vera L. Maria, Maria João Ribeiro, Sofia Guilherme, Amadeu M.V.M. Soares, Janeck Scott-Fordsmand, Mónica Amorim. Silver (nano)materials cause genotoxicity in Enchytraeus crypticus , as determined by the comet assay. Environmental Toxicology and Chemistry. 2017; 37 (1):184-191.

Chicago/Turabian Style

Vera L. Maria; Maria João Ribeiro; Sofia Guilherme; Amadeu M.V.M. Soares; Janeck Scott-Fordsmand; Mónica Amorim. 2017. "Silver (nano)materials cause genotoxicity in Enchytraeus crypticus , as determined by the comet assay." Environmental Toxicology and Chemistry 37, no. 1: 184-191.

Journal article
Published: 01 August 2016 in Marine Environmental Research
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The transcriptomic effects of Ibuprofen (IBU) in the digestive gland tissue of Mytilus galloprovincialis Lam. specimens exposed at low environmental concentrations (250 ng L(-1)) are presented. Using a 1.7 K feature cDNA microarray along with linear models and empirical Bayes statistical methods 225 differentially expressed genes were identified in mussels treated with IBU across a 15-day period. Transcriptional dynamics were typical of an adaptive response with a peak of gene expression change at day-7 (177 features, representing about 11% of sequences available for analysis) and an almost full recovery at the end of the exposure period. Functional genomics by means of Gene Ontology term analysis unraveled typical mussel stress responses i.e. aminoglycan (chitin) metabolic processes but also more specific effects such as the regulation of NF-κB transcription factor activity.

ACS Style

Vera Lúcia Maria; Mónica Amorim; Maria Bebianno; Francesco Dondero. Transcriptomic effects of the non-steroidal anti-inflammatory drug Ibuprofen in the marine bivalve Mytilus galloprovincialis Lam. Marine Environmental Research 2016, 119, 31 -39.

AMA Style

Vera Lúcia Maria, Mónica Amorim, Maria Bebianno, Francesco Dondero. Transcriptomic effects of the non-steroidal anti-inflammatory drug Ibuprofen in the marine bivalve Mytilus galloprovincialis Lam. Marine Environmental Research. 2016; 119 ():31-39.

Chicago/Turabian Style

Vera Lúcia Maria; Mónica Amorim; Maria Bebianno; Francesco Dondero. 2016. "Transcriptomic effects of the non-steroidal anti-inflammatory drug Ibuprofen in the marine bivalve Mytilus galloprovincialis Lam." Marine Environmental Research 119, no. : 31-39.

Journal article
Published: 09 October 2015 in International Journal of Environmental Research and Public Health
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The effects of nanomaterials have been primarily assessed based on standard ecotoxicity guidelines. However, by adapting alternative measures the information gained could be enhanced considerably, e.g., studies should focus on more mechanistic approaches. Here, the environmental risk posed by the presence of silver nanoparticles (Ag NM300K) in soil was investigated, anchoring population and cellular level effects, i.e., survival, reproduction (28 days) and oxidative stress markers (0, 2, 4, 6, 10 days). The standard species Folsomia candida was used. Measured markers included catalase (CAT), glutathione reductase (GR), glutathione S-transferase (GST), total glutathione (TG), metallothionein (MT) and lipid peroxidation (LPO). Results showed that AgNO3 was more toxic than AgNPs at the population level: reproduction EC20 and EC50 was ca. 2 and 4 times lower, respectively. At the cellular level Correspondence Analysis showed a clear separation between AgNO3 and AgNP throughout time. Results showed differences in the mechanisms, indicating a combined effect of released Ag+ (MT and GST) and of AgNPs (CAT, GR, TG, LPO). Hence, clear advantages from mechanistic approaches are shown, but also that time is of importance when measuring such responses.

ACS Style

Luís André Mendes; Vera Lúcia Maria; Janeck J. Scott-Fordsmand; Mónica J. B. Amorim. Ag Nanoparticles (Ag NM300K) in the Terrestrial Environment: Effects at Population and Cellular Level in Folsomia candida (Collembola). International Journal of Environmental Research and Public Health 2015, 12, 12530 -12542.

AMA Style

Luís André Mendes, Vera Lúcia Maria, Janeck J. Scott-Fordsmand, Mónica J. B. Amorim. Ag Nanoparticles (Ag NM300K) in the Terrestrial Environment: Effects at Population and Cellular Level in Folsomia candida (Collembola). International Journal of Environmental Research and Public Health. 2015; 12 (10):12530-12542.

Chicago/Turabian Style

Luís André Mendes; Vera Lúcia Maria; Janeck J. Scott-Fordsmand; Mónica J. B. Amorim. 2015. "Ag Nanoparticles (Ag NM300K) in the Terrestrial Environment: Effects at Population and Cellular Level in Folsomia candida (Collembola)." International Journal of Environmental Research and Public Health 12, no. 10: 12530-12542.

Journal article
Published: 14 August 2015 in International Journal of Environmental Research and Public Health
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The mechanisms of toxicity of Ag nanoparticles (NPs) are unclear, in particular in the terrestrial environment. In this study the effects of AgNP (AgNM300K) were assessed in terms of oxidative stress in the soil worm Enchytraeus crypticus, using a range of biochemical markers [catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), glutathione reductase (GR), total glutathione (TG), metallothionein (MT), lipid peroxidation (LPO)]. E. crypticus were exposed during 3 and 7 days (d) to the reproduction EC20, EC50 and EC80 levels of both AgNP and AgNO3. AgNO3 induced oxidative stress earlier (3 d) than AgNP (7 d), both leading to LPO despite the activation of the anti-redox system. MT increased only for AgNP. The Correspondence Analysis showed a clear separation between AgNO3 and AgNP, with e.g. CAT being the main descriptor for AgNP for 7 d. LPO, GST and GPx were for both 3 and 7 d associated with AgNO3, whereas MT and TG were associated with AgNP. These results may reflect a delay in the effects of AgNP compared to AgNO3 due to the slower release of Ag+ ions from the AgNP, although this does not fully explain the observed differences, i.e., we can conclude that there is a nanoparticle effect.

ACS Style

Maria J. Ribeiro; Vera L. Maria; Janeck J. Scott-Fordsmand; Mónica J. B. Amorim. Oxidative Stress Mechanisms Caused by Ag Nanoparticles (NM300K) are Different from Those of AgNO3: Effects in the Soil Invertebrate Enchytraeus Crypticus. International Journal of Environmental Research and Public Health 2015, 12, 9589 -9602.

AMA Style

Maria J. Ribeiro, Vera L. Maria, Janeck J. Scott-Fordsmand, Mónica J. B. Amorim. Oxidative Stress Mechanisms Caused by Ag Nanoparticles (NM300K) are Different from Those of AgNO3: Effects in the Soil Invertebrate Enchytraeus Crypticus. International Journal of Environmental Research and Public Health. 2015; 12 (8):9589-9602.

Chicago/Turabian Style

Maria J. Ribeiro; Vera L. Maria; Janeck J. Scott-Fordsmand; Mónica J. B. Amorim. 2015. "Oxidative Stress Mechanisms Caused by Ag Nanoparticles (NM300K) are Different from Those of AgNO3: Effects in the Soil Invertebrate Enchytraeus Crypticus." International Journal of Environmental Research and Public Health 12, no. 8: 9589-9602.

Validation study
Published: 01 August 2014 in Environmental Research
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Folsomia candida (Collembola) is a standard soil ecotoxicological species; effect assessment includes survival and reproduction as endpoints. In the present study, and for the first time, a range of oxidative stress biomarkers measurement was optimized and validated. The antioxidant capacity was measured by the activities of catalase (CAT), glutathione reductase (GR), glutathione-s-transferase (GST) and content of total glutathione (TG). The oxidative damage in the lipid membranes was estimated by lipid peroxidation (LPO) and metallothionein (MT) levels. The exposure included the essential and non-essential metals Cu and Cd, in LUFA 2.2 natural standard soil, using a series of sampling times along a 10 days period (0, 2, 4, 6 and 10 days). Exposure concentrations were selected based on their reproduction EC50 values, 60 and 1000 mg/kg soil DW, for Cd and Cu respectively. The protocols were optimized and results show that oxidative stress biomarkers can be successfully used in F. candida, this being highly relevant as complementary information to the mechanistic level. The selected sampling times gave a good indication of the markers dynamic and can be reduced/adapted in future testing. Results showed that both metals caused an increase in the MT levels after 6 days but Cd acted as a stronger oxidant agent compared to Cu, i.e. causing higher damage. In sum, Cd mobilized/activated more antioxidant enzymes, but the increased activities were not enough to prevent LPO. This study confirms that the oxidative stress caused by Cd is higher despite the use of same reproduction EC50 indicating that toxicity seems more reversible for Cu than for Cd. Among others, GST and MT would be a good selection of biomarkers for Cd effect.

ACS Style

Vera Lúcia Maria; Maria João Ribeiro; Mónica Amorim. Oxidative stress biomarkers and metallothionein in Folsomia candida - responses to Cu and Cd. Environmental Research 2014, 133, 164 -169.

AMA Style

Vera Lúcia Maria, Maria João Ribeiro, Mónica Amorim. Oxidative stress biomarkers and metallothionein in Folsomia candida - responses to Cu and Cd. Environmental Research. 2014; 133 ():164-169.

Chicago/Turabian Style

Vera Lúcia Maria; Maria João Ribeiro; Mónica Amorim. 2014. "Oxidative stress biomarkers and metallothionein in Folsomia candida - responses to Cu and Cd." Environmental Research 133, no. : 164-169.

Journal article
Published: 16 June 2014 in Science of The Total Environment
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The UV filters and musk fragrances have come into focus because these compounds are contained and increasingly used not only in sunscreen products but also in many products of daily use, such as cosmetics, skin creams, plastics or varnish. In view of this, the main objective of the present work was to develop and validate a method for the determination of three UV filters, two UV stabilizers and four musks in mussel samples (Mytilus galloprovincialis). The procedure combined a QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) extraction method with an analysis by gas chromatography–tandem mass spectrometry (GC–MS/MS). The methodology allowed the determination of target analytes at trace concentration levels (ng/g), with mean recoveries ranging from 91 to 112%. A monitoring study was conducted in four beaches in the Portuguese coast which are impacted by recreational activities and outflow of treated waste water effluents in rivers. The results are used to assess the occurrence of UV filters in comparison with UV stabilizers and musk fragrances which indicate other activities than bathing.

ACS Style

M. Picot Groz; Maria Jesus Martinez Bueno; D. Rosain; H. Fenet; C. Casellas; Catarina Guerreiro Pereira; Vera Lúcia Maria; Maria Bebianno; E. Gomez. Detection of emerging contaminants (UV filters, UV stabilizers and musks) in marine mussels from Portuguese coast by QuEChERS extraction and GC–MS/MS. Science of The Total Environment 2014, 493, 162 -169.

AMA Style

M. Picot Groz, Maria Jesus Martinez Bueno, D. Rosain, H. Fenet, C. Casellas, Catarina Guerreiro Pereira, Vera Lúcia Maria, Maria Bebianno, E. Gomez. Detection of emerging contaminants (UV filters, UV stabilizers and musks) in marine mussels from Portuguese coast by QuEChERS extraction and GC–MS/MS. Science of The Total Environment. 2014; 493 ():162-169.

Chicago/Turabian Style

M. Picot Groz; Maria Jesus Martinez Bueno; D. Rosain; H. Fenet; C. Casellas; Catarina Guerreiro Pereira; Vera Lúcia Maria; Maria Bebianno; E. Gomez. 2014. "Detection of emerging contaminants (UV filters, UV stabilizers and musks) in marine mussels from Portuguese coast by QuEChERS extraction and GC–MS/MS." Science of The Total Environment 493, no. : 162-169.

Journal article
Published: 30 June 2011 in Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology
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This study aimed to assess the antioxidant system potential and lipid peroxidative effects, in the gill and digestive gland of Mytilus galloprovincialis exposed to individual and binary mixtures of benzo(a)pyrene (BaP) and Cu for 7 days. Data demonstrated that in mussels exposed to BaP antioxidant enzymes (catalase--CAT, total glutathione peroxidase--tGPx, glutathione S-transferase--GST and glutathione reductase--GR) and lipid peroxidation (LPO) increased in the gill. On the contrary, in the digestive gland inhibitory antioxidant effects (superoxide dismutase-SOD, GR, metallothioneins-MT) and no changes in LPO levels were detected. Cu was also a potent oxidant agent since MT and LPO levels increased in mussel gill, despite no LPO effect in the digestive gland. For both single contaminants the organ specificity and distinct physiologic/metabolism roles were evident in terms of antioxidant capacity. Gill SOD inhibition, MT and GST unchanged was a result of "simple independent action" of exposure to BaP and Cu. "Interactions" in the binary mixtures, led to absence of changes in LPO effects. In the digestive gland, BaP and Cu interactions were also responsible for the GST and LPO enhancement (antagonistic effects). The current findings demonstrate the differences in antioxidant responses where the organ dependency highlights each contaminant particular mode of action. Generally, in the gill "non-interactive" effects occurred with the lowest Cu concentration while "interactions" exist for the mixture with the highest Cu concentrations. In the digestive gland, "interactions" and "no interaction" effects occurred in all the binary mixtures. Complex contaminant mixtures interact differently based on target tissue which may lead to an imbalance in the mussels health status.

ACS Style

V.L. Maria; M.J. Bebianno. Antioxidant and lipid peroxidation responses in Mytilus galloprovincialis exposed to mixtures of benzo(a)pyrene and copper. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 2011, 154, 56 -63.

AMA Style

V.L. Maria, M.J. Bebianno. Antioxidant and lipid peroxidation responses in Mytilus galloprovincialis exposed to mixtures of benzo(a)pyrene and copper. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology. 2011; 154 (1):56-63.

Chicago/Turabian Style

V.L. Maria; M.J. Bebianno. 2011. "Antioxidant and lipid peroxidation responses in Mytilus galloprovincialis exposed to mixtures of benzo(a)pyrene and copper." Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 154, no. 1: 56-63.