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.

Pancreatic stellate cells exhibit adaptation to oxidative stress evoked by hypoxia.

BACKGROUND INFORMATION: Pancreatic stellate cells play a key role in the fibrosis that develops in diseases such as pancreatic cancer. In the growing tumour, a hypoxia condition develops under which cancer cells are able to proliferate. The growth of fibrotic tissue contributes to hypoxia. In this study, the effect of hypoxia (1% O2 ) on pancreatic stellate cells physiology was investigated. Changes in intracellular free-Ca2+ concentration, mitochondrial free-Ca2+ concentration and mitochondrial membrane potential were studied by fluorescence techniques. The status of enzymes responsible for the cellular oxidative state was analyzed by quantitative reverse transcription-polymerase chain reaction, high-performance liquid chromatography, spectrophotometric and fluorimetric methods and by Western blotting analysis. Cell viability and proliferation were studied by crystal violet test, 5-bromo-2-deoxyuridine cell proliferation test and Western blotting analysis. Finally, cell migration was studied employing the wound healing assay. RESULTS: Hypoxia induced an increase in intracellular and mitochondrial free-Ca2+ concentration, whereas mitochondrial membrane potential was decreased. An increase in mitochondrial reactive oxygen species production was observed. Additionally, an increase in the oxidation of proteins and lipids was detected. Moreover, cellular total antioxidant capacity was decreased. Increases in the expression of superoxide dismutase 1 and 2 were observed and superoxide dismutase activity was augmented. Hypoxia evoked a decrease in the oxidized/reduced glutathione ratio. An increase in the phosphorylation of nuclear factor erythroid 2-related factor and in expression of the antioxidant enzymes catalytic subunit of glutamate-cysteine ligase, catalase, NAD(P)H-quinone oxidoreductase 1 and heme oxygenase-1 were detected. The expression of cyclin A was decreased, whereas expression of cyclin D and the content of 5-bromo-2-deoxyuridine were increased. This was accompanied by an increase in cell viability. The phosphorylation state of c-Jun NH2 -terminal kinase was increased, whereas that of p44/42 and p38 was decreased. Finally, cells subjected to hypoxia maintained migration ability. CONCLUSIONS AND SIGNIFICANCE: Hypoxia creates pro-oxidant conditions in pancreatic stellate cells to which cells adapt and leads to increased viability and proliferation.

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.

Concentration of 12 Metals and Metalloids in the Blood of White Stork (Ciconia ciconia): Basal Values and Influence of Age and Gender.

The white stork (Ciconia ciconia) is being increasingly used in biomonitoring programmes of environmental contaminants due to its growing population in Europe; however, studies on inorganic elements are scarce. The blood of 70 white storks was collected and analysed by inductively coupled plasma mass spectroscopy (ICP-MS) to determine the presence of the following elements: lead (Pb), mercury (Hg), arsenic (As), nickel (Ni), iron (Fe), zinc (Zn), copper (Cu), selenium (Se), manganese (Mn), chromium (Cr), cobalt (Co), and cadmium (Cd). Our main goals were to determine the mean concentrations of these elements in the blood and to study its association with age and gender. Mean concentrations were highest for Fe, followed by Zn, and lowest for Co and Cd. The metal levels were similar to the values referred in the literature for the same species from different locations. No statistically significant differences were found between males and females. Regarding age, statistically significant differences were observed for Ni, Cu, Se, Hg, and Pb between young and adult animals (except for Pb, values in adults were higher than in fledglings). Many element concentrations were correlated, with the strongest correlations between the pairs Hg-Se, Hg-As, and Fe-Zn, mainly in adults. This study provides the baseline data for a monitoring program based on white stork blood as a nondestructive sample.

Characterization of plasma cholinesterase in rabbit and evaluation of the inhibitory potential of diazinon.

Several studies indicate that more than one cholinesterase form may be present in the blood of mammals. In this study the predominant plasma cholinesterase activity, the physiological cholinesterase activity as well as cholinesterase sex-dependent changes in non-exposed individuals of rabbit have been established. Plasma cholinesterase was characterized using three substrates (acetylthiocholine iodide, propionylthiocholine iodide, and S-butyrylthiocholine iodide) and three cholinesterase inhibitors (eserine sulfate, BW284C51 and iso-OMPA). The results indicated that propionylthiocholine was the preferred substrate by plasma cholinesterase followed by acetylthiocholine and butyrylthiocholine, and the predominant enzymatic activity was acetylcholinesterase. Physiological plasma cholinesterase activity was 198.9 ± 5.8 nmol/min/ml for male and 205.2 ± 5.0 nmol/min/ml for female using acetylthiocholine as substrate. Thus, sex had no significant effect on the physiological cholinesterase activity (p>0.05). In addition, the in vivo and in vitro sensitivity of plasma cholinesterase to diazinon was also investigated. In rabbits exposed to single doses of diazinon (25 or 125 mg/kg) the higher inhibitions of plasma cholinesterase were reached 9h after oral administration (53% and 87% inhibition, respectively). Cholinesterase activity significantly recovered up to values similar to pre-administration between 3 and 7d depending on the administered dose and sex of the animals. Plasma cholinesterase activity decreased to 24%, 53% and 74% of the initial activity at 9h of in vitro exposure to 1.25, 3.13 and 6.25mg/l of diazinon, respectively, and it remained steadily depressed throughout the experimental period (10d). This study has demonstrated the sensitivity of cholinesterase activity in plasma of rabbits following both in vivo and in vitro exposure to sub-lethal concentrations of diazinon.

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.

Biomarker responses and lethal dietary doses of tau-fluvalinate and coumaphos in honey bees: Implications for chronic acaricide toxicity.

Evidence suggests that acaricide residues, such as tau-fluvalinate and coumaphos, are very prevalent in honey bee colonies worldwide. However, the endpoints and effects of chronic oral exposure to these compounds remain poorly understood. In this study, we calculated LC50 and LDD50 endpoints for coumaphos and tau-fluvalinate, and then evaluated in vivo and in vitro effects on honey bees using different biomarkers. The LDD50 values for coumaphos were 0.539, and for tau-fluvalinate, they were 12.742 in the spring trial and 8.844 in the autumn trial. Chronic exposure to tau-fluvalinate and coumaphos resulted in significant changes in key biomarkers, indicating potential neurotoxicity, xenobiotic biotransformation, and oxidative stress. The Integrated Biomarker Response was stronger for coumaphos than for tau-fluvalinate, supporting their relative lethality. This study highlights the chronic toxicity of these acaricides and presents the first LDD50 values for tau-fluvalinate and coumaphos in honey bees, providing insights into the risks faced by colonies.

Mineral and potentially toxic element profiles in the soil-feed-animal continuum: Implications for public, environmental, and livestock health in three pasture-based sheep farming systems.

Grazing livestock derive most of their mineral requirements from foraging. The presence of toxic elements in soils has become a significant concern for food safety and ecosystem services. Understanding the mineral content profiles in soil and forage is crucial for assessing animal health, predicting potential transfers of minerals or heavy metals into the food-chain, and assessing threats to the environment and human health. In this study, Na, Mg, P, K, Ca, Mn, Fe, Co, Cu, Zn, Se, As, Cd, Hg, and Pb were measured to determine the mineral status of three different pasture-based farming systems (with grazing sheep livestock) in a Spanish region of significant economic importance. A risk assessment evaluation of animal, environmental, and human health was performed on soil, forage, feed, serum, milk, and wool. Notably, traces of Pb, and As were identified in pastures in all farms. Our calculation of pollution indices revealed moderate levels of contamination by various elements, including Co, Cu, Zn, Se, As, Cd, Hg, and Pb. The two farms with more intense agrosystem practices showed a significant potential ecological risk, characterized by high soil levels of Hg and Cd. Animals from these farms also had high concentrations of these metals in wool. Although the target Hazard Quotient derived from milk consumption suggests that dairy products from this area are safe for consumption for adults, only milk from a dehesa farm (mix of woodland and pastureland) was free of potential health concerns related to Pb exposure. Our assessment of mineral profiles reveals a cohesive relationship between soil quality and derived animal products, particularly of the Merino sheep breeding and farming system. The results reveal the importance of adopting and reinforcing strategies to preserve dehesas as a sustainable and environmentally friendly agrosystem in the western Mediterranean region.

Noninvasive heavy metal pollution assessment by means of Iberian wolf (Canis lupus signatus) hair from Galicia (NW Spain): a comparison with invasive samples.

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.

Organochlorine residues in muscle of European eels (Anguilla anguilla) from four Spanish Mediterranean wetlands and coastal lagoons.

European eels (Anguilla anguilla) are an endangered species throughout their range, and chlorine organic compounds are some of the most important pollutants for marine species. Data on contaminants in eel stocks remain incomplete, so organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) in muscle of European eels from four Spanish Mediterranean ecosystems were analyzed. COPs are presents in eels from all areas, but some compounds are not detected: HCH α, ß and γ (lindane), endosulfan sulfate, heptachlor, and PCBs 28, 52 and 180. The high percentage of DDT 2,4' in eels from S'Albufera des Grau Natural Park, an ecosystem with good ecological status according to the Water Framework Directive, and the presence of PCBs in S'Albufereta Natural Reserve indicate the need to carry out further studies in the future. The results obtained can improve the management of this species in the studied areas.

Prevalence of burnout syndrome among veterinarians in Spain.

OBJECTIVE: Burnout is a syndrome of emotional distress common in health-care professions. The aim of this study was to assess the potential levels of burnout syndrome among veterinarians in Spain as well as different factors involved in its prevalence. SAMPLE: 282 Spanish veterinarians (57.4% women and 42.6% men). PROCEDURES: All data were obtained through an online survey. The instrument utilized for this purpose was the Maslach Burnout Inventory Human Services Survey (adapted to a Spanish-speaking population), with analyses performed using statistical software. Moreover, a sociodemographic questionnaire was utilized. RESULTS: Within the considered population, 23% suffered from burnout syndrome and the average scores were 29 points for emotional exhaustion, 14 points for depersonalization, and 35 points for personal accomplishment. A higher prevalence of burnout was associated with small animal clinicians (75.3%) and women (66.1%), in comparison to large animal clinicians (24.6%) and men (33.8%). CLINICAL RELEVANCE: This study showed for the first time that burnout syndrome is a current and highly relevant risk for Spanish veterinarians. The results obtained could help to raise awareness of this reality and the factors involved in the development or exacerbation of this syndrome.

Metal and metalloid concentrations in wild mammals from SW Europe: European hedgehog (Erinaceus europaeus) and badger (Meles meles).

In recent years, there have been increasing ecological and global concerns associated to Potentially Toxic Elements (PTEs). Thus, the relevance of wild mammals as biomonitors has been globally recognised. In the present study, Cd, Pb, Hg, Zn and As concentrations were quantified in European hedgehog and badger inhabiting SW Europe, and cumulative trends in relation to age and sex were evaluated. Liver and kidney samples were collected, mineralised and PTE content was determined by ICP-MS. Zn was the most abundant element quantified in both organs (239 and 89.8 mg kg-1 for hedgehogs and 179 and 164 mg kg-1 dw for badgers). In hedgehogs, very high Hg concentration were quantified (4.35 and 15.5 mg kg-1 dw in liver and kidney), and Cd was the most abundant for badgers (4.70 and 7.61 mg kg-1 dw in liver and kidney). Positive correlations were observed for the concentrations of PTE in the organs of both species. Age-dependence increased only Cd concentration, with levels in adult kidneys being significantly higher. In this study, European hedgehog and badger were used as biomonitors for the determination of PTEs to provide current reference values in relatively non-polluted areas of SW Europe, and to enhance the use of these species for future ecotoxicological studies.

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.

Evaluating the chronic effect of two varroacides using multiple biomarkers and an integrated biological response index.

There is mounting evidence that acaricides are among the most prevalent medicinal compounds in honey bee hive matrices worldwide. According to OCDE guideline No. 245 chronic lethal concentration of tau-fluvalinate (at concentrations ranging from 77.5 to 523.18 ppm), coumaphos (59.8 ppm) and dimethoate (0.7 ppm) were determined. The activity of the biomarkers acetylcholinesterase (AChE), carboxylesterase (CbE), glutathione S-transferase (GST), catalase (CAT) and malondialdehyde (MDA) was analysed and as they are implicated in neurotoxicity, biotransformation and antioxidant defences, these values were combined into an integrated biomarker response (IBR). There was enhanced AChE, CAT and GST activity in honey bees exposed to tau-fluvalinate, while dimethoate inhibited AChE activity. Both dimethoate and coumaphos inhibited CbE activity but they enhanced CAT activity and MDA formation. Our results highlight how these biomarkers may serve to reveal honey bee exposure to commonly used acaricides.

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.

Malnutrition and Increased Risk of Adverse Outcomes in Elderly Patients Undergoing Elective Colorectal Cancer Surgery: A Case-Control Study Nested in a Cohort.

BACKGROUND: Malnutrition increases worse outcomes during hospital admission for elective colorectal cancer (CRC) surgery in older adults. METHODS: This work was designed an observational, monocentric, case-control study nested in a cohort of patients undergoing elective surgery for CRC disease at the Hospital Universitario de la Ribera (HULR) (Alzira, Valencia, Spain) between 2011 and 2019. The study considered patients with a CONUT score in the range of moderate to severe malnutrition (>4 points), with control patients with normal nutritional situations or mild malnutrition. RESULTS: Moderate-to-severe malnutrition cases presented a greater length of stay (LOS), a higher incidence of adverse events (both medical and surgical complications), a higher incidence of surgical-wound infection, a greater need for blood transfusion, and a greater amount of transfused packed red blood cells. During hospitalization, the percentage of patients without nutritional risk decreased from 46 to 9%, and an increase in mild, moderate, and severe risk was observed. Patients with severe nutritional risk at hospital admission had significantly increased mortality at 365 days after discharge (HR: 2.96 (95% CI 1.14-7.70, p = 0.002)). After adjusting for sex, age, and Charlson index score, patients with severe nutritional risk at admission maintained a higher mortality risk (HR: 3.08 (95% CI 1.10-8.63, p = 0.032)). CONCLUSION: Malnutrition prevalence is high in older adults undergoing CRC elective surgery. Furthermore, this prevalence increases during hospital admission. Malnutrition is linked to worse outcomes, such as LOS, surgical and clinical complications, and mortality. For this reason, nutritional interventions are very important in the perioperative period.

Effects of carbofuran on the sea bass (Dicentrarchus labrax L.): study of biomarkers and behaviour alterations.

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.

Sex- and age-dependent accumulation of heavy metals (Cd, Pb and Zn) in liver, kidney and muscle of roe deer (Capreolus capreolus) from NW Spain.

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.