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.

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.

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.

Occurrence and ecological risk assessment of organic UV filters in coastal waters of the Iberian Peninsula.

This study aimed to assess the presence of 21 UVFs and metabolites in coastal regions of the Iberian Peninsula, to evaluate their environmental risk, and identify possible influential factors affecting their measured concentrations. Sampling was carried out in spring and summer to assess possible seasonal variations. UVFs were detected in 43 of the 46 sampling sites. Only 5 were found above LOD: BP4, OC, BP3 and metabolites BP1 and BP8. Samples collected in Mar Menor had the greatest variety of compounds per sample and the highest cumulative concentrations. The risk was characterized using Risk Quotients (RQ). BP1 showed a Low environmental Risk in 2 sites while for OC the RQ indicated a Moderate Risk in 22 points. The variables that contribute most to the variation are population density, sampling season, whether it was an open bay or not, and level of urbanization. The presence of WWTPs had a lower influence.

Bioaccumulation of CuO nanomaterials in rainbow trout: Influence of exposure route and particle shape.

The bioaccumulation potential of spherical and rod-shaped CuO nanomaterials (NMs) was assessed in rainbow trout (Oncorhynchus mykiss) exposed via water and diet following the OECD Test Guideline No. 305. Fish were exposed via diet to both NMs at concentrations of 70 and 500 mg Cu/kg for 15 days, followed by 44 days of depuration. For water-borne exposure, only the rod-shaped CuO NMs were tested at 0.08 and 0.8 mg Cu/L for 28 days, followed by 14 days of depuration. The concentration of Cu was determined in fish whole body to derive biomagnification and bioconcentration factors (BMF and BCF). Different tissues were sampled to investigate the total Cu biodistribution and target organs as well as the particle number-based bioaccumulation of CuO NMs. Estimated BMF and BCF values were below the thresholds of concern. However, shape and route influenced depuration. Following dietary exposure, there was a higher depuration of Cu from fish exposed to the rod-shaped compared to the spherical CuO NMs. A higher depuration was also observed for rod-shaped CuO NMs following the dietary exposure compared the aqueous one. Despite the much higher dietary exposure concentrations of rod-shape CuO NMs, similar Cu body burdens were reached via water. Cu was found in particulate form in different tissues. Although these NMs had a low bioaccumulation potential, differences in distribution and elimination patterns of Cu were observed depending on the exposure route and particle shape. Careful consideration of the most relevant exposure route is needed when designing a bioaccumulation experiment for testing NMs.

Occurrence and ecological risk assessment of antibiotic residues in urban wastewater discharged into the coastal environment of the Persian Gulf (the case of Bandar Abbas).

This is the first attempt to detect antibiotic residues released into the Persian Gulf marine environment. In particular, this study quantifies and assesses the ecological risks of amoxicillin (AMX) and azithromycin (AZM) residues in wastewater outfalls from Bandar Abbas, one of the major coastal cities in southern Iran. The wastewater effluent samples were collected from two main wastewater discharging stations, Gursuzan and Suru, between December 2020 and February 2021. High-performance liquid chromatography (HPLC) analysis revealed the average concentration (± 95% CL) of AMX were 460 ± 230.0 µg L-1 and 280 ± 100.6 µg L-1 in Gursuzan and Suru stations. Mean AZM concentrations were also 264 ± 10.59 µg L-1 and 295 ± 89.75 µg L-1 in these stations, respectively. Pooled data indicated that there are 335.17 ± 105.11 and 288.17 ± 37.94 µg of AMX and AZM residues in the wastewater per liter. The values of potential ecological risk, hazard quotient (HQ), were extensively above 10 (AMX: 90,586.5 and AZM: 5541.7) which suggest that these substances have a high health risk for the ecosystem and public. Given that Bandar Abbas wastewater treatment plant (WWTP) outlets are discharged at about 500 to 700 L s-1, the daily maximum potential AMX and AZM released were estimated to be 19.05 (± 0.283) × 103 and 14.74 (± 0.113) × 103 g day-1, respectively (α = 0.05). Our findings show that there is a concerning volume of antibiotic residues released into the northern Persian Gulf, and hence urgent policies and actions are necessary to reduce this pollution.

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.

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.

Melatonin modulates red-ox state and decreases viability of rat pancreatic stellate cells.

In this work we have studied the effects of pharmacological concentrations of melatonin (1 µM-1 mM) on pancreatic stellate cells (PSC). Cell viability was analyzed by AlamarBlue test. Production of reactive oxygen species (ROS) was monitored following CM-H2DCFDA and MitoSOX Red-derived fluorescence. Total protein carbonyls and lipid peroxidation were analyzed by HPLC and spectrophotometric methods respectively. Mitochondrial membrane potential (ψm) was monitored by TMRM-derived fluorescence. Reduced (GSH) and oxidized (GSSG) levels of glutathione were determined by fluorescence techniques. Quantitative reverse transcription-polymerase chain reaction was employed to detect the expression of Nrf2-regulated antioxidant enzymes. Determination of SOD activity and total antioxidant capacity (TAC) were carried out by colorimetric methods, whereas expression of SOD was analyzed by Western blotting and RT-qPCR. The results show that melatonin decreased PSC viability in a concentration-dependent manner. Melatonin evoked a concentration-dependent increase in ROS production in the mitochondria and in the cytosol. Oxidation of proteins was detected in the presence of melatonin, whereas lipids oxidation was not observed. Depolarization of ψm was noted with 1 mM melatonin. A decrease in the GSH/GSSG ratio was observed, that depended on the concentration of melatonin used. A concentration-dependent increase in the expression of the antioxidant enzymes catalytic subunit of glutamate-cysteine ligase, catalase, NAD(P)H-quinone oxidoreductase 1 and heme oxygenase-1 was detected in cells incubated with melatonin. Finally, decreases in the expression and in the activity of superoxide dismutase were observed. We conclude that pharmacological concentrations melatonin modify the redox state of PSC, which might decrease cellular viability.

Presence of pharmaceutical compounds, levels of biochemical biomarkers in seafood tissues and risk assessment for human health: Results from a case study in North-Western Spain.

This study assessed the presence of 27 pharmaceutically active compounds belonging to common therapeutic groups (cardiovascular, antiashmatic, psychoactive, diuretic, analgesic/anti-inflammatory, and antibiotic drugs) in the tissues of representative seafood species of bivalves, cephalopods, arthropods, and fish of high economic importance and consumption rates in North-Western Spain. Four pharmaceutical compounds, out of the 27 analyzed, were detected in the collected samples. The benzodiazepine citalopram was detected in the tissues of common octopus (14.1 ng g-1 dry weight) and pod razor (9.4 ng g-1 dw). The anxiolytic venlafaxine was detected in the tissues of common cockle (2.9 ng g-1 dw). The veterinary antiparasitic ronidazole was found in pod razor (2.3 ng g-1 dw) and, finally, the psychoactive compound alprazolam was also measured in common octopus (0.3 ng g-1 dw). Hazard quotients were calculated to assess the hazard posed by the consumption of the sampled seafoods. Octopus and pod razor tissues containing citalopram and alprazolam exceeded our chosen hazard limits (HQ > 0.1) for toddlers who are high consumers of seafood (HQ values between 0.18 and 0.27). A battery of biochemical biomarkers of effects (acetylcholinesterase, glutathione S-transferase; catalase, glutathione peroxidase and glutathione reductase enzymes activities and reduced/oxidized glutathione and malondialdehyde levels) was applied to samples of the study species with the aim of characterizing their basal levels and evaluating their suitability as a tool in the monitoring chronic exposure to environmental contaminants such as those analyzed in this study. According to the measured biomarkers, pod razor and cockles have the potential to be good bioindicator species, based on the observed among-site differences detected on acetylcholinesterase, glutathione S-transferase, catalase, glutathione peroxidase activities; reduced/oxidized glutathione and malondialdehyde levels.

Metal content in the liver, kidney, and feathers of Northern gannets, Morus bassanus, sampled on the Spanish coast.

The value of birds as bioindicators for monitoring the environmental inorganic elements has been globally recognized. In this context, due to its well-known ecology and population stability, the Northern gannet (Morus bassanus) could be particularly useful. Dead Northern gannets (n = 30) were collected and samples from the liver, kidney, and feathers were taken, dried, mineralized, and finally analyzed via ICP-MS. Metals and metalloids, namely As, Cd, Hg, Pb, and Zn, associated with environmental pollution and toxicity on living organisms, were evaluated. The mean highest concentrations of As, Hg, and Zn were found in the liver (0.916, 7.026, and 89.81 mg/kg dry weight, respectively). For Cd, the kidney showed the highest mean concentration (17.51 mg/kg dry weight), whereas for Pb, this value corresponded to the feathers (0.399 mg/kg dry weight). Significant differences were found between the age classes in terms of contaminant concentrations, with the adults exhibiting higher metal levels. This difference was significantly relevant for Pb and Hg, where the effect of age was observed for all the considered tissues. When considering the effect of gender, no significant differences were observed, in agreement with similar studies performed in other geographical regions. Finally, positive correlations between the concentrations of Hg and Pb in the feathers and in the liver (r = 0.688, p < 0.001 and r = 0.566, p < 0.001, respectively) were observed, as well as between the feather and kidney concentrations (r = 0.685, p < 0.001) indicating the possibility to use feathers, a non-invasive biomonitoring tissue, for better understanding Hg and Pb exposure in seabirds.

Melatonin induces reactive oxygen species generation and changes in glutathione levels and reduces viability in human pancreatic stellate cells.

In this study, the effects of pharmacological concentrations of melatonin (1 µM-1 mM) on human pancreatic stellate cells (HPSCs) have been examined. Cell type-specific markers and expression of melatonin receptors were analyzed by western blot analysis. Changes in intracellular free Ca2+ concentration were followed by fluorimetric analysis of fura-2-loaded cells. Reduced glutathione (GSH) and oxidized glutathione (GSSG) levels were determined by fluorescence techniques. Production of reactive oxygen species (ROS) was monitored following 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate acetyl ester and MitoSOX™ Red-derived fluorescence. Cell viability was studied using the AlamarBlue® test. Cultured cells expressed markers typical of stellate cells. However, cell membrane receptors for melatonin could not be detected. Thapsigargin, bradykinin, or melatonin induced changes in intracellular free Ca2+ concentration. In the presence of the indole, a decrease in the GSH/GSSG ratio was observed that depended on the concentration of melatonin used. Furthermore, the indole evoked a concentration-dependent increase in ROS production in the mitochondria and in the cytosol. Finally, melatonin decreased HPSC viability in a time and concentration-dependent manner. We conclude that melatonin, at pharmacological concentrations, induces changes in the oxidative state of HPSC. This might regulate cellular viability and could not involve specific plasma membrane receptors.

The organophosphorus pesticide dimethoate decreases cell viability and induces changes in different biochemical parameters of rat pancreatic stellate cells.

In the present study we employed cultured pancreatic stellate cells to study the effect of the organophosphorus insecticide dimethoate on pancreatic cell physiology. Esterase activity, cell viability, reactive oxygen species generation and Ca2+ mobilization were examined. Our results show that dimethoate (0.1, 1 and 10 µM) induced a concentration-dependent inhibition of cholinesterase enzymatic activity at all concentrations tested. A drop in carboxylesterase activity was noted in the presence of 10 µM dimethoate. In the presence of the pesticide a decrease in cell viability was detected. The clearer effect could be observed when the cells had been incubated during 96 h in the presence of dimethoate. The pesticide induced a slight but statistically significant increase in the production of reactive oxygen species in the mitochondria. Incubation of cells with dimethoate, in the presence of Ca2+ in the extracellular medium, led to a slow and progressive increase in [Ca2+]c towards an elevated value over the prestimulation level. A similar behavior was observed in the absence of extracellular Ca2+, indicating that dimethoate releases Ca2+ from the intracellular stores. Our results suggest that dimethoate might alter intracellular pathways that are critical for pancreatic physiology, creating a situation potentially leading to dysfunction in the exocrine pancreas.