Search

This work reports the development of an electrochemical biosensor for uric acid (UA), based on covalent immobilization of uricase (Uox) on cysteamine self-assembled monolayers (SAM). The monolayers were prepared over screen-printed gold electrode (AuSPE) modified with Au nanostructures for covalent linkage through EDC-NHS. The modified AuSPE was characterized by Scanning Electron Microscope (SEM) and Cyclic Voltammetry (CV). The results verified the cysteamine SAM formation, and the improved electrochemical performance of the working surface. The AuSPE/SAM/Uox biosensor presented a linear range of 0.84 – 16.81 mg/dL (50-1000 µM), a sensibility of 9.1423 µA/(mg/dL)cm2, and a limit of detection (LOD) of 0.0700 mg/dL. The biosensor did not present a response due to ascorbic acid (AA), verifying the selectivity of the assembly only to UA oxidation. The AuSPE/SAM/Uox showed a promising performance as a reliable, simple and fast detection tool for UA.

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 p p p p p 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.

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, The work was funded by the Consejería de Economía e Infraestructuras of Junta de Extremadura (Convocatoria de Proyectos de investigación en los centros públicos de I + D + i de la Comunidad Autónoma de Extremadura, 2016; Ref. IB16112); project code: 2017/00206/001.

9 Pág. Departamento de Medio Ambiente y Agronomía​, 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.

Zinc, cadmium, and lead in livers, kidneys, and feathers of 48 young and adult Atlantic puffins found dead or dying off the coast of Galicia (Northwest Spain) were determined. The most abundant bet...

Background This study was conducted to provide much needed information on the potential of low solubility, highly biodurable nanoparticles (NPs) (TiO2 and CeO2 NPs), to bioaccumulate in fish and to investigate the relationship between an engineered nanomaterials surface coating, uptake and biokinetics in vivo. Rainbow trout (Oncorhynchus mykiss) were fed diets spiked with uncoated or surface coated (polyethylene glycol (PEG) or citrate (CIT)) TiO2 and CeO2 NPs (100 mg NPs/kg feed) for 10 days and thereafter fish were allowed to depurate for 42 days. Special care was taken to measure the real dispersed and actual administered concentrations, taking into consideration any potential losses from leaching, and to characterise the form (size, aggregation state, charge) of the NPs to which the fish were exposed. Results The coatings had an influence on levels of uptake and distributions. Most notably a higher uptake of PEG and CIT coated TiO2 and CeO2 NPs compared to fish exposed to uncoated materials was observed. The elimination of any Ti from tissues was rapid during the first day of depuration, whereas Ce levels remained in fish tissues (stomach, intestine and liver) with differences in depuration and redistribution of the three types of CeO2 NPs. However, no bioaccumulation potential for both tested metal oxide NPs in fish irrespective of coatings is expected according to BMF values Conclusions Distinct uptake, distribution and depuration kinetics in rainbow trout have been evidenced for different metal oxide NPs (TiO2 and CeO2 NPs). Coatings influenced uptake and in some cases led to slower depuration and distinct distributions, but do not make the uncoated NPs studied bioaccumulative (BMF > 1). Graphical Abstract

10 Pág. Departamento de Medio Ambiente y Agronomía​ (INIA), 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., This work was cofinanced by the European Regional Development Fund (ERDF) and the Junta de Extremadura (GR 18080) and by the Junta de Extremadura through several research contracts.

8 Pág. Departamento de Medio Ambiente y Agronomía​, 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., The study was supported by grants IB18025 and GR18119 from Junta de Extremadura, Spain.

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.

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.

Nowadays, software projects and in particular open source ones heavily rely on a plethora of tools (e.g., Git, GitHub) to support and coordinate development activities. Despite their paramount value, they foster to fragment members' contribution, since members can access them with different identities (e.g., email, username). Thus, researchers and practitioners willing to evaluate individual members contributions are often forced to develop ad-hoc scripts or perform manual work to merge identities. This comes at the risk of obtaining wrong results and hindering replication of their work. In this demo we present SortingHat, which helps to track unique identities of project members and their related information such as gender, country and organization enrollments. It allows to manipulate identities interactively as well as to load bulks of identities via batch files (useful for projects with large communities). SortingHat is a component of GrimoireLab, an industry strong free platform developed by Bitergia, which offers commercial software analytics and is part of the CHAOSS project of the Linux Foundation. A video showing SortingHat is available at https://youtu.be/724I1XcQV6c.

The aim of this study was to determine the levels of different inorganic elements (lead [Pb], mercury [Hg], and arsenic [As]) and persistent chlorinated pollutants (including polychlorinated biphenyls [PCBs] and organochlorine pesticides [OCPs]) in blood and plasma of White stork (Ciconia ciconia) nestlings from northwest (NW) Spain. The concentrations of PCBs were lower than the limit of detection in all samples. The OCPs γ-HCH, 4,4'-DDE, HCB, and endosulfan were detected most frequently in plasma from White stork nestlings. These OCPs were detected in 98, 54, 39, and 37 % of all samples, respectively. However, the concentrations of organic pollutants were lower than the risk thresholds for birds. The mean levels of the inorganic elements Pb, Hg, and As were found to be 36.92 ± 33.48, 16.48 ± 12.87, and 9.813 ± 13.84 µg/L, respectively. These levels were also lower than the risk thresholds for birds. This study not only provides a snapshot of the levels of both inorganic and organic contaminants in wild White storks in NW Spain, it also provides a useful baseline for biomonitoring levels of the measured contaminants in this area.

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.

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.

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) 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.

Cd, Pb, and Zn were quantified in liver and kidney of red foxes (Vulpes vulpes) which were hunted during the 2003–2011 hunting seasons in Galicia (NW Spain). The effects of age and gender were evaluated to determine whether these variables should be included in future biomonitoring studies. The concentrations of hepatic and renal Cd (average 0.6 and 1.3 µg/g) and Pb (0.8 and 0.06 µg/g, respectively) were similar to background levels, with no known toxicological relevance. Similarly, the average levels of Zn in liver and kidney (77 and 17 µg/g) were in the range of physiological levels for canids. Although no significant gender-dependent variations were observed, the effect of aging was evident: the levels of hepatic Pb and both hepatic and renal Cd were higher in adults than in juveniles. Age should be included as a parameter during future biomonitoring programs focusing on trace metal bioaccumulation in red foxes.

The aim of this study was to determine the impact of specific environmental contaminants on the activities of antioxidant enzymes in terrestrial, free-living birds. To investigate this, several biomarkers and the levels of lead, mercury, and arsenic were determined in blood of white stork (Ciconia ciconia) nestlings from two different colonies located in the west of Spain that were exposed to different environmental conditions and human activities. Different biomarker and contaminant patterns were observed between the two colonies, suggesting different levels of environmental contamination. In comparison to nestlings from a natural reference area, the levels of the three hazardous elements Pb, As, and Hg were significantly higher in the colony nearby a landfill. Analysis of antioxidant biomarkers revealed significantly higher levels of glutathione and glutathione S-transferase activity in birds from the colony close to the landfill site than were found in those from the “control” area. In contrast, the le...

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.

The European Union FP-7 project GUIDEnano developed a web-based guidance tool, which guides users to assess human and environmental risks of nanomaterial-enabled products throughout their life cycle. One of the aims in the GUIDEnano hazard assessment strategy is to derive safety limit values based on existing human toxicity and ecotoxicological studies. Clear criteria needed to be established to select studies that could be used for such purpose. In the present paper, we present an approach for a systematical and quantitative evaluation of the quality of environmental and human toxicity studies performed with nanomaterials. The approach builds upon previous initiatives and includes refinements to facilitate its application by users with limited toxicological expertise. It covers in vivo and in vitro human toxicity studies as well as ecotoxicological studies addressing the toxicity to all environmental compartments. A scoring system related to test design and reporting considerations was developed following the principles of the Klimisch score (K score). In addition, the approach includes a scoring system based on the physicochemical properties that have been characterized and reported for the nanomaterial, including properties characterized in the exposure medium (S score). These two scores (K and S) are combined to obtain an overall quality score (Q score) that can be used to select studies, to weight different studies, and/or to introduce uncertainty factors in the risk assessment process. During its development, the approach has been tested and refined with 137 peer-reviewed articles. The final quality assessment approach and the results of its evaluation are presented here.

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.

Concentrations of 7 different polychlorinated biphenyl (PCB) congeners, and eleven organochlorine pesticides (OCPs) and metabolites, including DDTs (dichlorodiphenyltrichloroethane), HCHs (hexachlorocyclohexane isomers), Endosulfan, Endosulfan sulfate, Endrin, Dieldrin and HCB (hexachlorobenzene), were determined in adipose tissue of 57 yellow-legged gulls collected from NW and N Spain. Furthermore, the possible differences due to two endogenous factors, age and gender, were determined. All the analyzed PCBs were detected in over 66% of the samples, with levels of 291.9 (PCB 180), 34.5 (PCB 118), 0.7 (PCB 28), 432.6 (PCB 153), 225.5 (PCB 138), 1.3 (PCB 101) and 0.4 (PCB 52) µg/kg of adipose tissue. With respect to the OCPs and metabolites, only 4,4′-DDE and HCB were detected in more than 50% of the samples, with means of 360.6 and 2.5 µg/kg of adipose tissue, respectively. From all the considered contaminants, only 4,4′-DDE levels presented significant differences depending on the gender, with females showing higher values than males (p < 0.01). Significant differences (p < 0.001) were also found related to age for the levels of PCBs 180, 138, 101, 28 and 153, as well as 4,4′-DDE, with adult levels being higher than those in young birds. The results of the present study constitute a baseline to better assess the environmental impacts of PCB and OCP contamination at other coastal sites for future biomonitoring studies, with particular emphasis on gender- and age-related differences.

We investigated the levels of Zn(II], Pb(II] and Cd(II] in livers and muscles of the wild duck (Anas platyrhynchos) hunting in El Melah lagoon in northeastern Tunisia from December 2010. Analysis of variance shows that no significant differences in metals levels were found between samples, but it should be pointed out that the number of samples was small. HSD Tukey test show that the amount of Cd(II], Zn(II] and Pb(II] in the levers was higher than in the muscles. The results obtained suggest the importance of wild ducks as bioindicators of heavy metal pollution.

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 Caceres 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.

The present study is aimed to evaluate the effect of sublethal exposure to the pyrethroid pesticide deltamethrin, according to biometric parameters and important liver biomarkers in the common carp (Cyprinus carpio). Fish were exposed for 4, 15 and 30 days to deltamethrin mixed to the aquaria water at the concentrations of 0.08, 0.4 and 0.8μg/L. The exposure did not affect fish biometric parameters. Glutathione-S-transferase (GST) activity in liver (hepatopancreas) increased in fish exposed to the high concentration of pesticide only after 30 days when compared to controls. A significant increase of malondialdehyde (MDA) content (79.4% and 90.2% of control) was remarked after 15 and 30 days of exposure to the highest concentration of pesticide. Catalase activities (CAT) went up after 4 days of was to the concentration of 0.4μg/L and 0.8μg/L and at 15 days of exposure to 0.4μg/L. The most significant change of glutathione reductase (GR) activity was recorded at the fourth day, when GR activity was significantly increased after exposure to the concentrations of 0.4μg/L and 0.8μg/L of deltamethrin.

This study presents a consumer and farmer safety evaluation on the use of four botanical pesticides in pepper berry crop protection. The pesticides evaluated include preparations from clove, tuba root, sweet flag and pyrethrum. Their safety evaluation was based on their active ingredients being eugenol, rotenone, β-asarone and pyrethrins, respectively. Botanical pesticides from Acorus calamus are of possible concern because of the genotoxic and carcinogenic ingredient β-asarone although estimated margins of exposure (MOE) for consumers indicate a low priority for risk management. For the other three botanical pesticides the margin of safety (MOS) between established acute reference doses and/or acceptable daily intake values and intake estimates for the consumer, resulting from their use as a botanical pesticide are not of safety concern, with the exception for levels of rotenone upon use of tuba root extracts on stored berries. Used levels of clove and pyrethrum as botanical pesticides in pepper berry crop production is not of safety concern for consumers or farmers, whereas for use of tuba root and sweet flag some risk factors were defined requiring further evaluation and/or risk management. It seems prudent to look for alternatives for use of sweet flag extracts containing β-asarone.

The aim of this study was to determine heavy metal reference levels for risk assessment studies. For this purpose, the levels of lead, cadmium, copper and zinc were determined in liver tissues of wild boars sampled in NW Spain. The mean values were 0.383, 0.326, 23.50 and 56.86 mg/kg dried weight, respectively. In general, the levels detected were similar to or lower than the levels reported in literature. This study not only provides a useful baseline for biomonitoring the levels of the analyzed contaminants in wildlife in NW Spain, it also helps to understand the effects of gender on the levels of these elements. Similar to studies performed in other geographical regions, no significant gender-related differences could be detected. Although differences were not significant, the levels of zinc, cadmium and lead were modestly higher in males (55.78, 0.346 and 0.424 mg/kg, respectively) compared to females (45.25, 0.305 and 0.341 mg/kg). Our results indicate that, although gender did not significantly affect heavy metal uptake and toxicokinetics of contaminants in wild boars, these effects could vary between species, populations, organs, and elements. It is therefore essential to investigate gender-related differences for each species. © 2017 Elsevier Inc.

In the present study the effect of a chronic exposition to the organophosphorous pesticide diazinon on the porphyrin profile in feces of rabbit was evaluated, in order to validate the use of such molecules as non-destructive biomarkers for monitoring exposure of mammals to this environmental xenobiotic. Male and female (12:12) rabbits were exposed to oral single doses of diazinon, feces being sampled at every 10 days, till the end of the experience (30 days). HPLC method was validated from the results obtained, for detection of porphyrins in feces of mammals. Results revealed differences on the porphyrin profile between male and female, the most relevant differences associated to the uroporphyrin levels. In conclusion, porphyrin levels in rabbit's excreta may be used as indicators of exposure to such chemicals, thus providing a useful non-destructive method for monitoring exposure of animals to different environmental pollutants. However, the effect of gender should be taken into account when interpreting results.

The objective of this study was to investigate the acute effects of the pesticide carbofuran on the sea bass (Dicentrarchus labrax) using parameters at different levels of biological organisation (swimming behaviour and several biomarkers) and possible relationships between alterations found in different effect criteria. In a bioassay, sea bass juveniles were individually exposed to different doses of carbofuran (31, 63, 125 and 250 μg/L) for 96 h. At the end of the bioassay, the swimming performance and 11 biomarkers were determined. Biomarkers were: hepatosomatic index (HSI), lipid peroxidation (LPO), reduced glutathione and the activities of the enzymes ethoxyresorufin O-deethylase (EROD), glutathione S-transferases, superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, brain acetylcholinesterase (AChE) and muscle cholinesterases (ChE). After 96 h of exposure, carbofuran induced a decrease of the swimming velocity and inhibition of EROD activity at all concentrations tested, and inhibition of muscle ChE and brain AChE activities at 250 μg/L. No relevant alterations in any of the other tested parameters were found. These results show that carbofuran induced adverse effects on fish by interfering with neurofunction, capability of detoxication and swimming velocity. In addition, positive and significant correlations between the swimming velocity and (i) brain AChE activity, (ii) muscle ChE activity and (iii) EROD activity suggest that the inhibition of these enzymes may somehow be related to the behavioural changes observed. Since these functions are determinant for the survival and performance of the fish in the wild, the findings of the present study suggest that adverse effects may occur in populations exposed to carbofuran if a sufficient number of animals is affected.

In this study, the chronic effect of the insecticides carbofuran and deltamethrin on acetylcholinesterase (AChE) activity and malondialdehyde (MDA) and reduced glutathione (GSH) levels were examined in the brain of tench. Both pesticides were evaluated in two separate experiments, and animals were exposed in a continuous flow-system to three different concentrations of carbofuran (0, 10 and 100 microg/L) and deltamethrin (0, 0.0039 and 0.039 microg/L) for 60 days. After that period, animals were kept into pesticide-free water for other 30 days. In all cases, animals were sampled every 10 days all along the experience. AChE activity was significantly inhibited in fish exposed to 100 microg/L of carbofuran, during the first 30 days of exposition, returning to basal levels after this initial period. With respect to deltamethrin exposure, AChE activity was not significantly affected. When considering MDA levels, significant changes could only be detected during the recovery period for both pesticides, with a maximum of induction at 70 and 80 days, respectively associated to the highest dose of carbofuran and deltamethrin. Similarly, GSH levels varied all along the experience, with a maximum of significant increase at day 80 of exposition to the highest dose of both pesticides. This study shows that changes in AChE brain activity in tench can be used as a biomarker of early pesticide exposition in environmental monitoring programs, whereas MDA and GSH levels could be more associated to long-term expositions. The above results confirm and broaden former observations, suggesting that more investigations are needed before these biochemical parameters can be used as biomarkers.

This article describes a case of Chinaberry tree poisoning diagnosed in a dog. The initial clinical signs were variable and included tremors (muscular seizures) and a moderate limp in the dog's back leg, which evolved to a more severe condition in the following hours. Abdominal radiographic evaluation was requested, and abundant small, foreign, radio-dense bodies were detected, which were associated with Chinaberry tree fruits after surgical extraction. Adequate treatment was established, and the patient recovered completely. In addition, we compare clinical and gross postmortem findings in other similar cases reported in the literature. There is a general lack of information of such poisoning in pets.

The effect of a long-term exposure of tenchs to different concentrations (10 and 100 micro g/L) of the pesticide carbofuran has been evaluated. Microsomal hepatic cytochrome P450 subfamily 1A (CYP1A) and 3A (CYP3A) activities, as well as the phase II enzyme uridine diphospho-glucuronosyltransferase (UDPGT) activity were evaluated as adequate biomarkers of fish exposure to environmentally relevant concentrations of the pesticide carbofuran in freshwater ecosystems. A clear time-dependent inhibition of both CYP1A and UDPGT activities was observed in fish exposed to the highest dose of carbofuran with respect to controls, whereas in the case of CYP3A activity, values of exposed animals did not show a clear pattern of alteration during the experiment. The results of the present study demonstrated that hepatic CYP1A and UDPGT activities from tench could be considered as sensitive biomarkers for carbamate pesticides in polluted water, thus allowing future and ecologically relevant biomonitoring studies with this species.

Los Bifenilos Policlorados son una amplia familia de contaminantes ambientales persistentes de enorme relevancia ambiental, pues a pesar de estar prohibidos, siguen encontrándose en concentraciones preocupantes en el medio ambiente, suficientes como para desencadenar programas de biomonitorizacion ambiental. Y en esta necesidad de seguir identificándolos y cuantificándolos se encuentra, en un punto destacado, su potencial efecto como disruptores endocrinos, tema de suma importancia desde el punto de vista de la reproducción de las especies de vida salvaje. De hecho, en las dos últimas décadas, se ha incrementado entre la comunidad científica el interés por investigar las posibles alteraciones en el funcionamiento del sistema endocrino de vertebrados e invertebrados como consecuencia de la exposición a productos químicos. En definitiva, la presente revisión resume los conocimientos más actualizados existentes sobre los Bifenilos policlorados cuantificados en la fauna salvaje, y sus principales efectos sobre la reproducción. Les biphényles polychlorés sont une grande famille de polluants environnementaux persistants d'une énorme importance pour l'environnement, parce que bien qu'ils soient interdits, on trouve encore des concentrations alarmantes dans l'environnement suffisant pour déclencher des programmes de biosurveillance de l'environnement. Et ce besoin de continuer à identifier et quantifier eux est, dans un point culminant son effet potentiel comme perturbateurs endocriniens, question importante du point de vue de la reproduction des espèces d'animaux sauvages. En fait, au cours des deux dernières décennies, il a augmenté dans l'intérêt de la communauté scientifique enquêter sur d'éventuelles modifications dans le fonctionnement du système endocrinien des vertébrés et des invertébrés à la suite de l'exposition aux produits chimiques. En bref, la présente revue résume les connaissances de la date actuelle sur les polychlorobiphényles quantifiés dans la faune sauvage, et leurs principaux effets sur la reproduction. Polychlorinated biphenyls are a broad family of persistent environmental pollutants of enormous environmental significance, because despite being prohibited, they are still found in alarming concentrations in the environment, enough to trigger environmental biomonitoring programs. Associated to this need to continue identifying and quantifying them, their potential effect as endocrine disruptors is of great relevance, being an important issue from the point of view of the reproduction of wildlife species. In fact, in the last two decades, the scientific community interest about investigating the alterations in the functioning of the endocrine system of vertebrates and invertebrates because of exposure to chemicals has increased. In short, the present review summarizes current date knowledge on polychlorinated biphenyls quantified in wildlife, and their main effects on reproduction.

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. © 2016 Elsevier B.V.

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.

Los moluscos, debido a su capacidad filtrante, pueden acumular metales llegando a contaminar los alimentos y como consecuencia pueden tener una repercusion negativa sobre la salud de los consumidores. En este estudio se han determinado los contenidos de algunos metales (Cu, Zn, Cd y Pb) en muestras de almejas comercializadas en Galicia (Espana) tanto en conserva como en fresco. Para su analisis se ha empleado la voltamperometria de redisolucion anodica, obteniendose los niveles de dichos metales en musculo y en hepatopancreas. Los resultados han sido tratados estadisticamente para evaluar las posibles correlacciones en el contenido de metales en las distintas porciones anatomicas establecidas. Destacan las concentraciones mucho mas elevadas en hepatopancreas que en musculo para zinc, plomo y cobre en muestras en fresco. Asi, los maximos en hepatopancreas de almejas en fresco fueron para Zn: 1250 mg/kg; para Pb: 15,60 mg/kg y para Cu: 75,04 mg/kg, frente a los maximos de Zn (60,36 mg/kg), Pb (0,67 mg/kg) y Cu (9,48 mg/kg), en musculo. En cuanto al cadmio, en general las muestras en conserva presentaron concentraciones mas elevadas que las muestras en fresco, llegando a alcanzar el hepatopancreas de una de ellas hasta 8,11 mg/kg. Teniendo en cuenta la importancia de estos moluscos en la alimentacion y de acuerdo con la legislacion vigente, se ha podido establecer, desde el punto de vista toxicologico, que los niveles de estos metales no suponen riesgo para el consumidor. © 2005 Altaga. Todos los derechos reservados. Articulo en Espanol

Living beings are constantly exposed to a wide variety of environmental contaminants and their biochemical response can be employed to assess the possible biological impact of such pollution. Sex can play a major role in the uptake, fate, and effects of contaminants in organisms. The present study evaluated endogenous levels of different biomarkers in both male and female roe deer in order to determine background levels of these biomarkers and sex-related differences in this species. Post-mitochondrial hepatic, renal, and pulmonary cytochrome P450 subfamily 1A1, as well as phase II enzymes glutathione S-transferase and uridine diphospho-glucuronosyltransferase, and lipid peroxidation in terms of malondialdehyde levels were determined. The highest values of all the considered parameters were quantified in liver. Results showed no differences in levels of enzymatic activities in the three considered tissues. However, a statistical difference (P

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

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 min-1 per mg of protein; p, Istražen je utjecaj pesticida metomila na razine enzimskih aktivnosti u tkivima šarana izloženih subletalnoj koncentraciji od 0,34 mg L-1 tijekom 15 dana. Nakon 4 i 15 dana u jetrima i škrgama pokusnih životinja izmjerene su aktivnosti katalaze i glutation S-transferaze (GST), a aktivnost acetilkolinesteraze (AChE) u mozgu i mišićima. Aktivnost katalaze u jetrima blago se smanjila u izloženih riba u usporedbi s kontrolnom skupinom, no to je smanjenje bilo statistički značajno tek na početku pokusa. Istovremeno nisu zamijećene značajne promjene u aktivnosti katalaze u škrgama izloženih i kontrolnih riba (srednje vrijednosti bile su u rasponu 10,7-11,7 nmol min-1 po miligramu proteina. Dok je aktivnost enzima GST u jetrima izloženih riba bila tek blago inhibirana na početku pokusa, u škrgama je utvrđena značajna inhibicija. Aktivnost acetilkolinesteraze u mozgu s vremenom se smanjivala i pokazala značajan pad nakon 96 sati izloženosti (0,041 vs. 0,075 nmol min-1 po miligramu proteina; p

The effect of orally administered atrazine (25 or 100 mg/kg on days 0, 5, and 10 of the experiment) was studied in European quail (Coturnix coturnix coturnix) on four non-destructive biomarkers fecal porphyrins, blood glutathione-S-transferase, glutathione reductase, reduced glutathione, and malondialdehyde (MDA). Uroporphyrin I (UPI) and coproporphyrins I and III (CPIII) were the main porphyrins detected in feces. The lowest dose of ATZ caused a significant (P 〈 0.05) increase in UPI and CPIII at day 5, and the highest dose of ATZ caused an induction of CPI and a significant (P 〈 0.05) decrease in MDA levels at day 30. © Springer Science+Business Media New York 2013.

The current study represents the first investigation of the suitability of wolf hair as indicator tissue for metal exposure and accumulation within NW Spanish ecosystems. Dead animals, which were not specifically killed for these purposes, were necropsied, and further toxicological analyses were performed in order to establish the heavy metal content (Pb, Cd, and Zn) in liver, kidney, and hair, by means of inductively coupled plasma optical emission spectrometry. Two different factors, gender and age, were considered in order to determine their influence on heavy metal accumulation. Mean liver, kidney and hair concentrations of both toxic element, Cd (0.528, 2.692, and 0.026 ppm) and Pb (4.108, 0.031, and 0.196 ppm) considered on a dry weight basis, were situated below the established as acute toxicity levels for mammals. The highest concentrations were quantified for Zn, with means ranging from a maximum in hair samples (150.9 ppm) to a minimum in kidney samples (25.81 ppm). When the variable of gender and age were considered, female and adult wolves had higher concentrations of Cd in all the considered organs, although differences were only statistically significant for kidney. Neither the sex nor the age had a significant effect on Pb and Zn concentrations. A significant positive correlation (p < 0.05) was identified between hair and liver Pb concentrations, as well as between hair and kidney Cd concentrations, thus suggesting the interest of the noninvasive sample for future ecotoxicological biomonitoring studies.

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.

In the present study, bioaccumulation of lead, cadmium and zinc in liver, kidney and muscle of roe deer (Capreolus capreolus) from Galicia (NW Spain) was quantified. Metal analysis was performed by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). In general the quantified concentrations were associated with background environmental levels, except for Pb, where some markedly elevated concentrations were obtained, but with no toxicological relevance. The effect of sex on Cd, Pb and Zn levels was considered, females showing a general trend to accumulate higher concentrations of these metals, with the exception of the hepatic content of Cd (which was significantly higher in males, P < 0.05) and Pb levels, which did not differ according to sex. The effect of age was analysed only in male animals and was clearly observed for Pb levels, with concentrations significantly higher in adult animals (liver: P < 0.01, kidney: P < 0.05 and muscle: P < 0.001) and for Cd levels in kidney samples (P < 0.001). This study provides the first data on heavy metal content in roe deer from the area of study and suggests that this species could be a good bioindicator of metal pollution, even though both factors, sex and age, could represent an important source of variation in the bioaccumulation of these metals in wild populations.