Toxic Effects of Different Coating-Related Functionalized Nanoparticles on Aquatic Organisms.

The peculiar physico-chemical characteristics of nanomaterials (NMs) and the use of different coatings to improve their expected properties result in a huge amount of nanoforms, which vary in chemical composition, size, shape and surface characteristics. This makes it almost impossible to test all the nanoforms available, and efforts have been made to establish grouping or read-across strategies. The aim of this work was to find a behavior pattern of effect among nanoforms of different metallic core nanoparticles (NPs) (TiO2, CeO2 and Ag NP) with the same coatings (sodium citrate, poly (ethylene glycol), dodecylphosphonic acid or oleylamine). Daphnia magna, rainbow trout and two fish cell lines (PLHC-1 and RTH-149) were exposed to a range of concentrations (up to 100 mg/L) of the uncoated or coated NPs. Ag NPs were the most toxic, followed by CeO2 NPs and finally by TiO2 NPs. The results show that a clear pattern of toxicity in the studied species could not be established related to the coatings. However, it was possible to confirm different inter-species sensitivities. RTH-149 was the most sensitive cell line, and Daphnia magna was more sensitive than fish. Moreover, some differences in coating-core interactions were found between the metal oxide and the metal NPs in Daphnia magna.

Biochemical Effects of Heavy Metals and Organochlorine Compounds Accumulated in Different Tissues of Yellow-Legged Gulls (Larus Michahellis).

In the present study, livers, kidneys and adipose tissue of Yellow-legged Gull (Larus michahellis) were collected. Samples were used to determine relationships between heavy metals/metalloids in liver and kidneys (Hg, Cd, Pb, Se and As) or persistent organic pollutants in adipose tissue (7 PCBs and 11 organochlorine pesticides) with biomarkers of oxidative stress (CAT, GPx, GR, GSH, GST, MDA) analysed in both internal organs. Three possible influencing variables have been studied: age, sex and sampling area. As a result, statistically significant differences (P < 0.05, P < 0.01) were only found according to the sampling area, with differences among the three studied areas found in both organs. Significant positive correlations (P < 0.01) were found in liver (Hg vs. GST; Se vs. MDA) and in kidney (As vs. GR; As vs. GPx; PCB52 vs. CAT; PCB138 vs. CAT). The scarcity in correlations suggests that the levels of pollutants found in animals were not high enough to trigger an effect at the oxidative level.

Use of poisoned baits against wildlife. A retrospective 17-year study in the natural environment of Extremadura (Spain).

This study reports the results obtained from toxicological analyses of different types of baits referred to the laboratory of the Toxicology Area (Faculty of Veterinary Medicine, Cáceres, Spain) over a 17-year period (2002-2018). These baits were suspicious materials found in the environment of the region of Extremadura (Western Spain), where such malpractices are a problem to be addressed, as wide livestock farming and hunting activities are combined with a significant wealth of wildlife (especially birds of prey). A total of 246 baits, including 32 commercial chemical products to be used in baits, were analysed. Samples from 183 cases were received and classified according to the material used for their preparation and the toxic substance found. Overall, the most common bait consisted of meat preparations (56.3% of cases) intended to eliminate predators considered 'annoying' for livestock and hunting practices, such as carnivores and scavengers. It should be noted that contact baits (as fenthion-impregnated perches) were also detected (7.6%). Regarding the substances detected, anticholinesterase compounds (organophosphates and carbamates) were the most commonly used substances for the preparation of baits (detected in 85.3% of positive baits). Moreover, 8% of the positive baits presented more than one toxic substance in their composition. Due to the types of toxic compounds and the methods used to prepare the baits, this study shows that the malicious use of highly toxic substances in the environment to kill wildlife is a common and current issue and poses a serious risk to different species.

Heavy metals and metalloid levels in the tissues of yellow-legged gulls (Larus michahellis) from Spain: sex, age, and geographical location differences.

In the present study, mercury (Hg), cadmium (Cd), lead (Pb), selenium (Se), and arsenic (As) were measured in liver, kidney, and feathers of adult, juvenile, and chick seagulls (Larus michahellis) collected from the northwest of Spain. Age, sex, and the geographical location of samples were considered variables that can influence metal bioaccumulation, for which concentrations were determined by means of ICP-MS. The mean concentrations (dry weight) found in seagulls were 7.01 ± 0.37 mg Hg/kg, 22.82 ± 2.83 mg Cd/kg, 7.36 ± 1.36 mg Pb/kg, 18.64 ± 0.63 mg Se/kg, and 10.64 ± 0.59 mg As/kg. Regarding the different factors analyzed, Hg was the only metal showing sex-related differences, being significantly higher (p < 0.05) the concentrations found in feathers of males (1.26 ± 0.12 mg/kg) than those in females (0.99 ± 0.11 mg/kg). A highly significant (p < 0.01) increase in levels of some metals was found in liver related to the increase of age: Hg (adults (A) 3.33 ± 0.22 mg/kg vs chicks (C) 1.76 ± 0.28 mg/kg), Cd (A 4.74 ± 0.62 mg/kg vs C 1.79 ± 0.2), Pb (A 0.65 ± 0.12 mg/kg vs juveniles 0.4 ± 0.11 mg/kg), and Se (A 7.56 ± 0.43 mg/kg vs C 5.24 ± 0.53 mg/kg). Positive correlations between Cd-Hg and Se-Hg were found in liver (p < 0.001), kidney (p < 0.001), and feathers (p < 0.05 and p < 0.001, respectively). The associations found may reflect antagonistic interactions between Se and Cd on Hg toxicity. The results suggest that L. michahellis can reveal local contamination around the foraging and breeding sites and can be a very useful monitoring instrument for assessing heavy metal contamination and sentinel species of environmental health.

Nanopharmaceuticals (Au-NPs) after use: Experiences with a complex higher tier test design simulating environmental fate and effect.

The current environmental hazard assessment is based on the testing of the pristine substance. However, it cannot be excluded that (nano)pharmaceuticals are excreted into sewage during the use phase followed by entry into wastewater treatment plants (WWTPs). Sorption to sewage sludge or release via effluent can result in modified ecotoxicological effects which possibly can only be detected with a modified test approach. The objective of our study was to investigate a realistic exposure scenario for metallic nanoparticles (NPs) in pharmaceutical products, excreted into effluent, and released into the environment after treatment in WWTPs. The test approach was illustrated by using gold (Au) NPs. Effluent from model WWTPs were investigated in aquatic tests (Daphnia magna, fish cell lines). Sewage sludge was used as a sole food source (Eisenia fetida) or mixed with soil and used as test medium (soil microorganisms, Folsomia candida, Enchytraeus crypticus). To cover the aspect of regulation, the test systems described in OECD-test guidelines (OECD TG 201, 211, 220, 232, 249, 317) were applied. Modifications and additional test approaches were included to meet the needs arising out of the testing of nanomaterials and of the exposure scenarios. The results were assessed regarding the suitability of the test design and the toxicity of Au-NPs. Except for activated sludge as a sole food source for E.fetida, the selected test approach is suitable for the testing of nanomaterials. Additional information can be gained when compared to the common testing of the pristine nanomaterials in the standardized test systems. Effects of Au-NPs were observed in concentrations exceeding the modeled environmental.

Heavy Metals Content in Great Shearwater (Ardenna Gravis): Accumulation, Distribution and Biomarkers of Effect in Different Tissues.

The purpose of this study was to explore the usefulness of Great Shearwater (Ardenna gravis) as a bioindicator for biomonitoring programs for metal pollution. Three different metals were analysed in liver, kidney, and feathers, including cadmium, lead, and zinc. Glutathione-S-transferase, malondialdehyde, reduced glutathione, and catalase were assessed as oxidative stress biomarkers. Sex-related trends in metal accumulation also were evaluated. In liver and kidney, the mean concentrations of Zn (146.1 ± 5.14 and 108 ± 2.70 mg/kg, respectively) and Pb (0.19 ± 0.01 and 0.13 ± 0.01 mg/kg, respectively) in A. gravis were generally comparable to values reported in other studies. However, animals presented slightly higher concentrations of Cd (9.67 ± 0.65 in liver and 17.41 ± 0.84 mg/kg in kidney) than those reported in the same species sampled in Southern Atlantic waters. The slightly higher levels of Cd found in this study compared with other studies are probably affected by the location in Northern Atlantic waters (with different diet intake). In feathers, levels of Zn (70.70 ± 1.76 mg/kg) were lower than in other Ardenna shearwaters, whereas higher levels were found for Cd (0.16 ± 0.01 mg/kg) and Pb (0.84 ± 0.06 mg/kg). The lack of differences found between males and females could be influenced by the migration status, because both sexes stay in similar physiological conditions, with no laying eggs. Levels found in the feathers of the present study were related to concentrations in internal tissues below those which cause adverse effects in birds. Thus, feathers would appear as a potential noninvasive tool for metals biomonitoring in seabirds, because it is possible to quantify them. Baseline data of oxidative stress levels have been reported, both in liver and kidney, presenting no correlations with the levels of metals in these tissues. The low internal metal levels and the lack of correlations between oxidative stress metrics suggest a low risk of the environmental concentrations for seabirds.

Monochloramine effects on gallbladder contractility.

Digestive inflammatory processes induce motility alterations associated with an increase in reactive oxygen species production, including monochloramine (NH2 Cl). The aim of the study was to characterize the effects of the naturally occurring oxidant monochloramine in the guinea pig gallbladder. We used standard in vitro contractility technique to record guinea pig gallbladder strips contractions. NH2 Cl caused a concentration-dependent contraction which was reduced by inhibition of extracellular Ca2+ influx and tyrosine kinase pathways. The PKC antagonist GF109203X also reduced the response but not after previous tyrosine kinase inhibition, suggesting that PKC is activated by tyrosine kinase activity. The NH2 Cl contractile effect was also reduced by inhibitors of mitogen-activated protein kinase (MAPK), nitric oxide synthase, phospholipase A2 and cyclooxygenase. In addition, NH2 Cl impaired the responses to CCK, tissue depolarization and electrical field stimulation. In conclusion, we present new evidence that monochloramine impairs not only the gallbladder response to CCK but also to membrane depolarization and nervous plexus stimulation, and that tyrosine kinase, PKC, MAPK and NO pathways are involved in the contractile direct effect of monochloramine.

Brain biomarkers in tench ( Tinca tinca L.) after semi-static exposure to the pesticide carbofuran / Biomarcadores cerebrales en tenca (Tinca tinca L.) tras una exposición semiestática al pesticida carbofurano / Biomarcadores cerebrais na tenca (Tinca tinca L.) após uma exposição semi-estática ao pesticida carbofurano

Abstract Background: The aquatic environment is continuously being contaminated with toxic chemical substances from industrial, agricultural and domestic activities. The identification of different biomarkers of such contamination is of great relevance to determine the potential impact of those xenobiotics on the health of organisms. Objectives: To estimate the suitability of different biochemical brain biomarkers from tench in order to evaluate the environmental effect of an exposition to different concentrations of the pesticide carbofuran. Methods: The present study focused on the effect of a long-term exposure (28 days) to different concentrations (50, 100 and 200 μg/L) of the pesticide carbofuran on certain biochemical parameters of tench (Tinca tinca L.). Selected biomarkers for stress monitoring were malondialdehyde (MDA), as index of lipid peroxidation, and reduced glutathione (GSH). Moreover, acetylcholinesterase (AChE) activity was quantified as biomarker of effect on fish nervous system. Results: AChE activity was significantly inhibited in fish exposed to all concentrations of carbofuran after 14 and 21 days of exposure, returning to basal levels after this period with 50 and 100 μg/L. Moreover, significant decreases of MDA levels could be detected after 14 and 21 days of exposure to 200 μg/L, lasting until the end of the experiment. GSH levels decreased after 14 and 21 days of exposure to 100 and 200 μg/L; nevertheless, GSH recovered basal levels at the end of the experiment. Conclusion: All the three biochemical parameters assessed -mainly inhibition of AChE activity- could be adequate biomarkers of early exposition (less than 14 days) to low doses of carbofuran in biomonitoring programs, being MDA reduction also related to longer exposure.

Resumen Antecedentes: El ambiente acuático está continuamente contaminado con sustancias químicas tóxicas provenientes de actividades industriales, agrícolas y domésticas. La identificación de diferentes biomarcadores de dicha contaminación es de gran relevancia para determinar el impacto potencial de esos xenobióticos en la salud de los organismos. Objetivo: Estimar la idoneidad de diferentes biomarcadores cerebrales bioquímicos de tenca para evaluar el efecto ambiental de una exposición a diferentes concentraciones del pesticida carbofurano. Método: El presente estudio se centró en los efectos de una exposición a largo plazo (28 días) a diferentes concentraciones (50, 100 y 200 μg/L) del pesticida carbofurano sobre determinados parámetros bioquímicos de la tenca (Tinca tinca L.). Los biomarcadores seleccionados para el control del estrés fueron el malondialdehído (MDA), como indicador de peroxidación lipídica, y el glutatión reducido (GSH). Además, se cuantificó la actividad acetilcolinesterasa (AChE), como biomarcador de efecto sobre el sistema nervioso del pez. Resultados: La actividad AChE se inhibió significativamente en los peces expuestos a todas las concentraciones de carbofurano después de 14 y 21 días de exposición, regresando a los niveles basales tras este periodo en las exposiciones con 50 y 100 μg/L. Así mismo se detectaron descensos significativos de los niveles de MDA tras 14 y 21 días de exposición con 200 μg/L, manteniéndose hasta el final del experimento. Los niveles de GSH descendieron tras 14 y 21 días de exposición con 100 y 200 µg/l; sin embargo, los niveles basales se recuperaron al final del experimento. Conclusión: Los tres parámetros bioquímicos evaluados, pero principalmente la inhibición de la actividad de AChE, podrían usarse como biomarcadores de exposición temprana (menos de 14 días) a bajas dosis de carbofurano en programas de biomonitorización, estando la reducción de MDA también relacionada con exposiciones más prolongadas.

Resumo Antecedentes: O meio aquático está sendo continuamente contaminado com substâncias químicas tóxicas de atividades industriais, agrícolas e domésticas. A identificação de diferentes biomarcadores de tal contaminação é de grande relevância para determinar o impacto potencial desses xenobióticos na saúde dos organismos. Objetivo: Estimativa da idoneidade de várias biomarcadores cerebrais bioquímicos de tenca para avaliar o efeito ambiental de uma exposição a diferentes concentrações do pesticida carbofurano. Método: O presente estudo centrou-se no efeito de uma exposição a longo prazo (28 dias) a diferentes concentrações (50, 100 e 200 μg/L) do pesticida carbofurano em certos parâmetros bioquímicos da tenca (Tinca tinca L.). Os biomarcadores selecionados para o monitoramento do estresse foram malondialdeído (MDA), como índice de peroxidação lipídica e glutationa reduzida (GSH). Além disso, a atividade da acetilcolinesterase (AChE) foi quantificada, como biomarcador de efeito sobre o sistema nervoso do peixe. Resultados: A atividade de AChE foi significativamente inibida em peixes expostos a todas as concentrações de carbofurano após 14 e 21 dias de exposição, voltando aos níveis basais após esse período com 50 e 100 μg/L. Além disso, as diminuições significativas dos níveis de MDA podem ser detectadas após 14 e 21 dias de exposição a 200 μg/L, até o final do experimento. Os níveis de GSH diminuíram após 14 e 21 dias de exposição a 100 e 200 μg/L, porém GSH recuperou os níveis basais no final do experimento. Conclusão: Todos os três parâmetros bioquímicos avaliados, mas principalmente a inibição da atividade AChE, podem ser biomarcadores adequados de exposição precoce (menos de 14 dias) a baixas doses de carbofurano em programas de biomonitorização, sendo a redução de MDA também relacionada a exposições mais longas.

Acute toxic effects caused by the co-exposure of nanoparticles of ZnO and Cu in rainbow trout.

The toxic effects produced by the co-exposure to low- and non-toxic concentrations of zinc oxide (ZnONPs) and copper nanoparticles (CuNPs) was assessed in rainbow trout following the OECD Test Guideline 203. Four groups of trouts were exposed for 96 h to a range of concentrations (0.0425-0.34 mg/L) of CuNPs (50 nm) in combination with a fixed non-toxic concentration (1.25 mg/L) of ZnONPs (25 nm) determined from an independent concentration-response study. One additional group was exposed to the highest concentration of CuNPs alone. Behaviour and mortality were observed during the experiment. After 96 h exposure, accumulated levels of Cu and Zn in the fish were measured by ICP-MS and ICP-OES, respectively. The induction of oxidative stress in liver and gills was evaluated by the glutathione-S-transferase (GST) activity and the reduced glutathione (GSH) / oxidized glutathione (GSSG) ratio. The ethoxyresorufin-O-deethylase (EROD) activity was also assessed. The results showed that CuNPs at the highest tested concentration do not cause acute toxicity, whereas exposure to all mixtures caused mortality, which was inversely proportional to the concentration of CuNPs (from 28% to 86% survival). Accumulated levels of Cu and Zn in the fish increased with the increasing concentrations of CuNPs, suggesting that the presence of CuNPs favours the entry of Zn. In general, the GST activity increased significantly in the gills of co-exposed groups, whereas the GSH/GSSG ratio was altered in the liver. The EROD activity was not modified. In conclusion, the co-exposure to these NPs potentiates their toxicity, observing an alteration of the GST activity and GSH/GSSG ratio in gill and liver, which was more pronounced at the lowest concentration of CuNPs. The lower toxic effect observed with the highest concentrations of CuNPs coincides with a greater internalization of Zn.

Mercury (Hg), Lead (Pb), Cadmium (Cd), Selenium (Se), and Arsenic (As) in Liver, Kidney, and Feathers of Gulls: A Review.

Mercury (Hg), lead (Pb), cadmium (Cd), selenium (Se), and arsenic (As) are metals or metalloids of high concern because of their effects on the environment and, specially, their potential toxicity on the animals inhabiting there. Due to their relevance, these elements have been object of several biomonitoring studies in different animal species around the world. Birds are widespread and, as species, are able to supply specific and relevant information about the regions where they live, being useful as bioindicators, as long as they are not birds with a strong migratory character. The main goal of this review is to summarize data collected from different studies using seabirds, paying special attention to gulls, in order to be helpful for coming studies and regulatory affairs.Several tissues have been used to evaluate Hg, Cd, Pb, Se, and As concentrations in seabirds, being focused the present review in those analyzing the liver, kidneys, and feathers. The most frequently analyzed tissue for Hg was the liver, followed by feathers, and finally kidney. For Cd levels, most of the studies were carried out in the liver, followed by feathers and kidneys. Pb, Se, and As levels were determined to a lesser extent. Feathers should be taken carefully as indicator of accumulation of pollutants, since procedure during analysis may lead to controversial results.Some authors reported that interspecific differences in the exposure of elements are determined by multiple factors, including properties of the contaminant, species, feeding habits, migratory status, sex, and age.The present review provides a comprehensive overview of the analyzed elements' occurrence in different species of seabirds, including gulls. Therefore, it can be a useful database providing for Hg, Pb, Cd, Se, and As levels in different tissues of seabirds.

Acute hazard of biocides for the aquatic environmental compartment from a life-cycle perspective.

One of the aims of the European project LIFE-COMBASE is to build a computational tool to predict the acute toxicity for aquatic organisms of biocidal active substances and its environmental degradation products. A database was implemented compiling toxicity data for these substances in organisms of the freshwater/marine and sewage treatment plant compartments. The goal of this study is to analyze the compiled data to identify the possible hazard of these compounds for the aquatic compartments. Several official and scientific databases were consulted. Data from 196 biocidal substances and 206 environmental metabolites were collected for the taxonomic groups, including fish, invertebrates, algae and sewage treatment plant (STP) microorganisms. Substances were categorized for their toxicity in four groups, considering values of L(E)C50, according to EU Regulation (EC) No 1272/2008. >50% of the parent were located in category 1 (L(E)C50 ≤ 1 mg/L) for fish, invertebrates and algae, indicating a high toxicity for the freshwater/marine compartments. However >60% were not toxic for STP microorganisms. Metabolites were mainly less toxic than the parent compounds, but 22-36% presented the same toxicity and ~6% were more toxic. No toxicological information was found for ~50% of the metabolites for fish, invertebrates and algae, reaching the 96% for the microorganisms. In addition, information on toxicity to the STP microorganisms was only found for 40% of the parent compounds. The high percentage of toxic metabolites and the scarcity of data for these compounds indicate the need to further study their impact in the aquatic compartments.

Influence of sex on biomarkers of oxidative stress in the kidney, lungs, and liver of rabbits after exposure to diazinon.

The present study evaluated the effect of two oral doses of the pesticide diazinon (25 and 125 mg/kg bw) on four biochemical parameters (malondialdehyde and glutathione levels, and catalase and glutathione S-transferase enzymatic activities) in the kidney, lungs, and liver of rabbit after 10 days of exposure. Malondialdehyde levels were significantly greater in exposed animals compared to controls, especially in the animals exposed to the higher dose of the pesticide. This result was reflected in the glutathione levels, which were significantly lower at that same higher dose. Catalase activity was also inhibited by the higher dose of the pesticide in all three organs analysed, whereas inhibition of glutathione S-transferase activity was only significant for the kidney and lungs. There were some slight differences between the sexes: taking the results for all three organs, the higher dose of diazinon resulted in a clearly significant inhibitory effect on the catalase activity and glutathione levels in males, and a significant enhancing effect on the malondialdehyde levels in females. These results help to confirm the interest of considering such endogenous factors in future ecotoxicological studies, and that the four biomarkers considered are suitable for reflecting the toxic effects of diazinon on rabbits, especially the effects related to oxidative stress. Graphical abstract.

Association of In Utero Persistent Organic Pollutant Exposure With Placental Thyroid Hormones.

In utero exposure to persistent organic pollutants (POPs) can result in thyroid function disorder, leading to concerns about their impact on fetal and neonatal development. The associations between placental levels of various POPs and thyroid hormones (THs) were investigated. In a prospective Danish study initially established for assessing congenital cryptorchidism, 58 placenta samples were collected from mothers of boys born with (n = 28) and without (n = 30) cryptorchidism. The concentrations of polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins/furans (PCDD/Fs), organotin chemicals (OTCs), organochlorine pesticides (OCPs), T4, T3, and rT3 were measured. The associations between placental THs and various POPs were analyzed using multiple linear regression. Five PBDEs, 35 PCBs, 14 PCDD/Fs, 3 OTCs, 25 OCPs, T4, T3, and rT3 were measured. No correlation between THs and the odds of cryptorchidism was found. Several POPs were significantly associated with THs: (1) T4 was inversely associated with BDEs 99, 100, ΣPBDE, and 2378-TeCDD, and positively associated with 1234678-HpCDF; (2) T3 was positively associated with 2378-TeCDF and 12378-PeCDF; and (3) rT3 was positively associated with PCB 81, 12378-PeCDF, and 234678-HxCDF, and inversely associated with tributyltin, ΣOTC, and methoxychlor. These results revealed that POP exposures were associated with TH levels in placenta, which may be a possible mechanism for the impacts of POP exposures on children's growth and development. This study provides new insight into the complexity of thyroid-disrupting properties of POPs. More research is needed to elucidate the biological consequences of POP exposures.

Levels of perfluorinated acids (PFCAs) in different tissues of Lepidochelys olivacea sea turtles from the Escobilla beach (Oaxaca, Mexico).

Lepidochelys olivacea is the most abundant and globally distributed sea turtle species in the world and thus, monitoring this species for persistent organic pollutants, such as perfluorinated chemicals, is fundamental for their protection. This study was the first to evaluate the occurrence of five PFCAs (PFOA, PFNA, PFDA, PFUnA, PFDoA) in liver and blood samples of Olive Ridley turtle population from the Escobilla beach (Oaxaca, Mexico). PFDA and PFUnA were the predominant PFCs in blood samples (detected in 93% and 84% of samples, respectively) and were also present in the highest concentrations. Liver samples showed higher PFCA concentrations than whole blood samples, with PFNA and PFDA the most abundant PFCs congeners in liver samples, detected in 65% and 47% of the samples, respectively. The measured levels of contaminants in the blood samples of Lepidochelys olivacea sea turtles were compared to the levels reported in the literature for other turtle species. While linear significant correlations between PFNA, PFDA and PFUnA concentrations in blood samples and curved carapace lengths were determined, no correlation was found for PFOA, supporting the hypothesis that sea turtles could have a higher ability to eliminate this perfluorinated chemical from their blood than other PFCAs. However, we do not know if the concentrations are species or sampling areas dependent.

Mechanisms underlying the enhancement of toxicity caused by the coincubation of zinc oxide and copper nanoparticles in a fish hepatoma cell line.

Ecosystems are exposed to a wide variety of individual substances, including at the nano-scale; and the potential adverse effects of their interactions are an increasing concern. The purpose of the present study was to determine whether zinc oxide nanoparticles (ZnONPs) at a no-observed-effect concentration modulate the cytotoxicity of copper nanoparticles (CuNPs) in the fish hepatoma cell line PLHC-1 after 48 h of exposure and the contribution of the released ions to these effects. Cells were exposed to 50-nm CuNPs (0.39-25.0 µg/mL), alone or in combination with ZnONPs (25 nm or 100 nm), at 6.25 µg/mL. Cells were exposed to suspensions of NPs or to their supernatants, as well as to their combinations. The effects on cell viability were assessed through cytotoxicity assays. Changes in cell morphology and metal internalization were also evaluated. The cytotoxicity exerted by CuNPs was enhanced in the presence of nontoxic concentrations of ZnONPs. On the contrary, Zn ions protected the cell line from the CuNP toxicity, this effect being related to an increase in the intracellular levels of Zn. This increase of metal was not observed in cells exposed to both ZnONPs and CuNPs, even when they were visualized inside the cell. The results indicated that the internalization of ZnONPs, but not the Zn ions, was responsible for the enhanced toxicity of the CuNPs. Environ Toxicol Chem 2016;35:2562-2570. © 2016 SETAC.

Chlorinated pollutants in blood of White stork nestlings (Ciconia ciconia) in different colonies in Spain.

The aim of this study was to investigate the levels of persistent chlorinated pollutants (POPs) in wild birds. The concentrations of multiple POPs, including polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) were determined in plasma of White stork nestlings. Blood samples were collected from three breeding colonies located in the West of Spain that were exposed to different environmental conditions and human activities. The concentrations of PCBs were below the limit of detection in all samples. The OCPs Heptaclor, 4,4'-DDE, endosulfan and aldrin were detected most frequently in plasma from White stork nestlings. From these four compounds, 4,4'-DDE and Heptaclor were found in high abundance. OCPs could not be detected in the colony breading nearby a landfill and an intensive agricultural area, indicating that these banned compounds are not taken up by crops growing in this area. However, birds from the colony breading next to a landfill exhibited the highest OCPs levels. These high levels might be due to exposure of the mothers to OCPs during their migration to Africa and subsequent transfer to their eggs.

Different enzymatic activities in carp (cyprinus carpio L.) as potential biomarkers of exposure to the pesticide methomyl.

This study investigated the influence of the pesticide methomyl on different enzymatic activities in carp. The fish were exposed to a sub-lethal concentration (0.34 mg L-1) of methomyl for 15 days. On days 4 and 15, catalase (CAT) and glutathione-S-transferase (GST) activities were measured in the liver and gills. Acetylcholinesterase (AChE) activity in brain and muscle was also determined. Liver catalase activity slightly increased in exposed fish when compared to controls, but it was statistically significant only at the beginning of the experiment. No changes in CAT activity in the gills of exposed and control animals were observed (mean values were in the range 10.7-11.7 nmol min-1 per mg of protein). Liver GST activity was slightly inhibited in the exposed animals at the beginning of the study; however, it was significantly inhibited in the gills. Brain AChE activity was diminished throughout the experiment and significantly decreased after 96 h of exposure compared to controls (0.041 vs. 0.075 nmol min1 per mg of protein; p<0.001). Our findings suggest that CAT, GST, and AChE are reliable biomarkers of effect after exposure to methomyl.

Breeding near a landfill may influence blood metals (Cd, Pb, Hg, Fe, Zn) and metalloids (Se, As) in white stork (Ciconia ciconia) nestlings.

Cadmium, lead, mercury, selenium, iron, zinc and arsenic levels were measured in blood samples from 59 free-ranging white stork nestlings from colonies located in three different environmental conditions in Western Spain. The reference colony was situated in "Llanos de Cáceres y Sierra de Fuentes", an Area of Special Interest for Bird Protection. A second colony was located close to (4.9 km) an urban landfill and a third one was close to both an intensive agricultural area and an urban landfill (1.5 km). Blood samples were diluted and elemental analysis was performed using inductively coupled plasma mass spectrometer. In all cases, the essential metals zinc and iron were found at the highest mean concentrations followed by lead > selenium > mercury > arsenic > cadmium. Regarding toxic metals, the highest concentrations were found for lead (ranging from 23.27 to 146.4 µg/L) although in all cases the concentrations were lower than those considered to cause subclinical effects. The metals levels detected in the chick's blood were not related to the previously reported levels in the soil next to the colonies, which may indicate that landfills are the main source of metals in white stork nestlings. The present data showed that metal levels in white stork chicks may be influenced by the use of landfills as feeding areas by the parents. However, more studies on the metal content in the feed of white stork and the influence of the distance to the landfill are necessary to establish the causality of these findings.

Effects of carbofuran and deltamethrin on acetylcholinesterase activity in brain and muscle of the common carp.

This work investigated the effect from exposure to insecticides carbofuran and deltamethrin on acetylcholinesterase (AChE) activity in the brain and muscle of common carp (Cyprinus carpio). Both pesticides were evaluated through two separate experiments, and carp were exposed in a semi-static system to three different concentrations of carbofuran (10, 50, and 100 µg/L) and deltamethrin (0.08, 0.4, and 0.8 µg/L) during a month with sampling times at 0, 4, 15, and 30 days (n = 7 from each aquarium). AChE activity was significantly inhibited in both organs of carps exposed to carbofuran at all sampling times depending on dose and time, reaching inhibition values of 73.5 and 67.1%, in brain and muscle tissues respectively, after 30 days with the highest concentration. On the contrary, AChE activity was not significantly affected after deltamethrin exposure at all concentrations and times of the assay. This study shows that the measurement of brain and muscle AChE activity in Cyprinus carpio is a useful biomarker of carbamates exposure and/or effects, but has no application with pyrethroids.