Transgenerational effects of triazole fungicides on gene expression and egg compounds in non-exposed offspring: A case study using Red-Legged Partridges (Alectoris rufa).

Triazole fungicides are widely used to treat cereal seeds before sowing. Granivorous birds like the Red-legged Partridge (Alectoris rufa) have high exposure risk because they ingest treated seeds that remain on the field surface. As triazole fungicides can act as endocrine disruptors, affecting sterol synthesis and reproduction in birds several months after exposure, we hypothesized that these effects could also impact subsequent generations of exposed birds. To test this hypothesis, we exposed adult partridges (F0) to seeds treated at commercial doses with four different formulations containing triazoles as active ingredients (flutriafol, prothioconazole, tebuconazole, and a mixture of the latter two), simulating field exposure during late autumn sowing. During the subsequent reproductive season, two to four months after exposure, we examined compound allocation of steroid hormones, cholesterol, vitamins, and carotenoids in eggs laid by exposed birds (F1), as well as the expression of genes encoding enzymes involved in sterol biosynthesis in one-day-old chicks of this F1. One year later, F1 animals were paired again to investigate the expression of the same genes in the F2 chicks. We found changes in the expression of some genes for all treatments and both generations. Additionally, we observed an increase in estrone levels in eggs from partridges treated with flutriafol compared to controls, a decrease in tocopherol levels in partridges exposed to the mixture of tebuconazole and prothioconazole, and an increase in retinol levels in partridges exposed to prothioconazole. Despite sample size limitations, this study provides novel insights into the mechanisms of action of the previously observed effects of triazole fungicide-treated seeds on avian reproduction with evidence that the effects can persist beyond the exposure windows, affecting unexposed offspring of partridges fed with treated seeds. The results highlight the importance of considering long-term chronic effects when assessing pesticide risks to wild birds.

Toxicological effects assessment for wildlife in the 21st century: Review of current methods and recommendations for a path forward.

Model species (e.g., granivorous gamebirds, waterfowl, passerines, domesticated rodents) have been used for decades in guideline laboratory tests to generate survival, growth, and reproductive data for prospective ecological risk assessments (ERAs) for birds and mammals, while officially adopted risk assessment schemes for amphibians and reptiles do not exist. There are recognized shortcomings of current in vivo methods as well as uncertainty around the extent to which species with different life histories (e.g., terrestrial amphibians, reptiles, bats) than these commonly used models are protected by existing ERA frameworks. Approaches other than validating additional animal models for testing are being developed, but the incorporation of such new approach methodologies (NAMs) into risk assessment frameworks will require robust validations against in vivo responses. This takes time, and the ability to extrapolate findings from nonanimal studies to organism- and population-level effects in terrestrial wildlife remains weak. Failure to adequately anticipate and predict hazards could have economic and potentially even legal consequences for regulators and product registrants. In order to be able to use fewer animals or replace them altogether in the long term, vertebrate use and whole organism data will be needed to provide data for NAM validation in the short term. Therefore, it is worth investing resources for potential updates to existing standard test guidelines used in the laboratory as well as addressing the need for clear guidance on the conduct of field studies. Herein, we review the potential for improving standard in vivo test methods and for advancing the use of field studies in wildlife risk assessment, as these tools will be needed in the foreseeable future. Integr Environ Assess Manag 2024;20:699-724. © 2023 His Majesty the King in Right of Canada and The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC). Reproduced with the permission of the Minister of Environment and Climate Change Canada. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Wildlife ecological risk assessment in the 21st century: Promising technologies to assess toxicological effects.

Despite advances in toxicity testing and the development of new approach methodologies (NAMs) for hazard assessment, the ecological risk assessment (ERA) framework for terrestrial wildlife (i.e., air-breathing amphibians, reptiles, birds, and mammals) has remained unchanged for decades. While survival, growth, and reproductive endpoints derived from whole-animal toxicity tests are central to hazard assessment, nonstandard measures of biological effects at multiple levels of biological organization (e.g., molecular, cellular, tissue, organ, organism, population, community, ecosystem) have the potential to enhance the relevance of prospective and retrospective wildlife ERAs. Other factors (e.g., indirect effects of contaminants on food supplies and infectious disease processes) are influenced by toxicants at individual, population, and community levels, and need to be factored into chemically based risk assessments to enhance the "eco" component of ERAs. Regulatory and logistical challenges often relegate such nonstandard endpoints and indirect effects to postregistration evaluations of pesticides and industrial chemicals and contaminated site evaluations. While NAMs are being developed, to date, their applications in ERAs focused on wildlife have been limited. No single magic tool or model will address all uncertainties in hazard assessment. Modernizing wildlife ERAs will likely entail combinations of laboratory- and field-derived data at multiple levels of biological organization, knowledge collection solutions (e.g., systematic review, adverse outcome pathway frameworks), and inferential methods that facilitate integrations and risk estimations focused on species, populations, interspecific extrapolations, and ecosystem services modeling, with less dependence on whole-animal data and simple hazard ratios. Integr Environ Assess Manag 2024;20:725-748. © 2023 His Majesty the King in Right of Canada and The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC). Reproduced with the permission of the Minister of Environment and Climate Change Canada. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Assessing Contamination Profiles in Livers from Road-Killed Owls.

Raptors are recognized as valuable sentinel species for monitoring environmental contaminants owing to their foraging behavior across terrestrial and aquatic food webs and their high trophic position. The present study monitored environmental contaminants in livers from road-killed owls to evaluate differences in the exposure patterns due to factors such as species, age, and sex of individuals. Carcasses of road-killed individuals of eagle owl (Bubo bubo), long-eared owl (Asio otus), little owl (Athene noctua), tawny owl (Strix aluco), and barn owl (Tyto alba) were collected in Alentejo (Portugal). Eighty-one organic contaminants were analyzed, including organochlorine pesticides (OCPs), per- and polyfluoroalkyl substances (PFAS), polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), pharmaceuticals, in-use pesticides, and organophosphate esters (OPEs). Overall, 21 contaminants were detected. In all species ∑OCPs were prevalent at concentrations from 3.24 to 4480 ng/g wet weight, followed by perfluorooctane sulfonic acid (PFOS), the only PFASs detected (from 2.88 to 848 ng/g wet wt) and ∑PCBs (1.98-2010 ng/g wet wt); ∑PAHs were ubiquitous but detected at the lowest concentrations (7.35-123 ng/g wet wt). Differences among species were observed according to principal component analysis. Eagle owl and long-eared owl presented the highest levels of ∑OCPs, ∑PCBs, and PFOS, consistent with its higher trophic position, while ∑PAHs prevailed in tawny owl, barn owl, and little owl, related to their frequent use of urban areas for nesting and roadsides for hunting. Adults presented higher concentrations of ∑OCPs and ∑PCBs than juveniles, while no differences were observed for PFOS and ∑PAHs. Pharmaceuticals, in-use pesticides, and OPEs were not detected. Overall, the present study shows specific contamination patterns in five species with similar diet but with differences in habitat preferences. Environ Toxicol Chem 2024;43:821-832. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

HEALTH ASSESSMENT OF CAPTIVE BEARDED VULTURES (GYPAETUS BARBATUS) IN CATALONIA (SPAIN).

Bearded vulture (Gypaetus barbatus) populations are declining worldwide primarily due to anthropogenic factors. A captive breeding program has been established in Spain, a country with one of the largest free-living populations in Europe, to further enhance the conservation efforts of this emblematic species. However, captive vulture populations can be exposed to different risks through food, such as drug residues and antimicrobial-resistant (AMR) bacteria. Health surveillance of species involved in captive breeding programs is important to face introduction of healthy animals in situ and to obtain baseline clinical data. The objective of this study was to assess the general health status of bearded vultures held in captivity in Catalonia (northeastern Spain) by carrying out hematologic, biochemical, toxicologic, and bacteriologic analyses. A total of 16 bearded vultures were sampled; the data obtained from one vulture, with a chronic tibiotarsal fracture, were excluded from the statistical analysis. Hematologic and biochemical parameters of the bearded vultures were mostly within the range of standard values as stated in previous studies. Basal feather and serum corticosterone levels were analyzed and described for the first time in this species. A total of 15 Escherichia coli isolates were obtained that were resistant to fluoroquinolones (80%), tetracycline (60%), trimethoprim and ampicillin (40%), sulfamethoxazole (33%), and colistin (20%), with 40% of them being multidrug resistant. Three of 15 isolates were carriers of the mcr-1 gene. Only the injured bird previously treated with enrofloxacin was positive for fluoroquinolone residues. Periodic monitoring for the presence of AMR bacteria would be recommended in captive breeding programs as a preventive action to establish future therapies.

Diet and Spatial Ecology Influence Red-Legged Partridge Exposure to Pesticides Used as Seed Treatment.

Seed treatment with pesticides is an extended agricultural practice with a high risk to granivorous birds that consume those seeds. To characterize that risk, it is necessary to understand the ecological factors that determine the exposure chances of birds to treated seeds. We investigated how pesticide uptake by red-legged partridges was related to cultivated plant ingestion and to the use of recently sown fields. We analyzed pesticide residues in 144 fecal samples from 32 flocks and determined the plant diet composition using DNA metabarcoding. Habitat use was studied through the monitoring of 15 GPS-tagged partridges. We confirmed, through the analysis of seeds, that >80% of cereal fields from the area had seeds treated with triazole fungicides. Tebuconazole was detected in 16.6% of partridges' feces. During the sowing season, cultivated plants accounted for half of the plant diet, but no association was found between cultivated plant consumption and pesticide intake. GPS tracking revealed that tebuconazole was detected in feces when partridges had recently used sown fields, whereas nonexposed partridges showed no overlap with recently sown areas. Our results highlight the need to incorporate field ecology into the characterization of pesticide exposure to improve the efficacy of environmental risk assessment.

Medicated livestock carcasses and landfill sites: Sources of highly toxic veterinary pharmaceuticals and caffeine for avian scavengers.

Veterinary drugs are of concern in terms of potential environmental pollution and their negative impacts on avian scavengers. These pharmaceuticals reach vultures through the consumption of carcasses of previously treated livestock. Here, we analysed samples from livestock carcasses (n = 159), avian scavenger tissues (n = 116) and plasma (n = 312) for 49 compounds commonly used in veterinary medicine in Aragon (NE Spain) and nearby regions. Samples were analysed using liquid chromatography with electrospray ionization mass spectrometry (LC-ESI-MS/MS). We detected pharmaceuticals in 54.1% of livestock carcasses analysed (50.3% with antibiotics, 10.8% with NSAIDs). For veterinary pharmaceuticals in tissues and plasma from avian scavengers, we detected pharmaceuticals in 51.7% and 28.5% of samples, respectively. Antibiotics were detected in 50.9% and 25.3% while NSAIDs were determined in 6.0% and 5.5% of tissues and plasma from avian scavengers, respectively. Moreover, caffeine was detected in plasma in 73.7% of vultures sampled at landfill sites, indicating its usefulness as a biomarker of urban garbage ingestion. We found an association between livestock carcasses, especially pigs and chickens, and the presence of veterinary pharmaceuticals in avian scavengers. We highlight that carcass disposal for feeding avian scavengers must address the potential risks posed by veterinary pharmaceutical residues.

A heavy burden: Metal exposure across the land-ocean continuum in an adaptable carnivore.

Urbanisation and associated anthropogenic activities release large quantities of toxic metals and metalloids into the environment, where they may bioaccumulate and threaten both wildlife and human health. In highly transformed landscapes, terrestrial carnivores may be at increased risk of exposure through biomagnification. We quantified metallic element and metalloid exposure in blood of caracals (Caracal caracal), an adaptable felid inhabiting the rapidly urbanising, coastal metropole of Cape Town, South Africa. Using redundancy analysis and mixed-effect models, we explored the influence of demography, landscape use, and diet on the concentration of 11 metals and metalloids. Although species-specific toxic thresholds are lacking, arsenic (As) and chromium (Cr) were present at potentially sublethal levels in several individuals. Increased use of human-transformed landscapes, particularly urban areas, roads, and vineyards, was significantly associated with increased exposure to aluminium (Al), cobalt (Co) and lead (Pb). Foraging closer to the coast and within aquatic food webs was associated with increased levels of mercury (Hg), selenium (Se) and arsenic, where regular predation on seabirds and waterbirds likely facilitates transfer of metals from aquatic to terrestrial food webs. Further, several elements were linked to lower haemoglobin levels (chromium, mercury, manganese, and zinc) and elevated levels of infection-fighting cells (mercury and selenium). Our results highlight the importance of anthropogenic activities as major environmental sources of metal contamination in terrestrial wildlife, including exposure across the land-ocean continuum. These findings contribute towards the growing evidence suggesting cities are particularly toxic areas for wildlife. Co-exposure to a suite of metal pollutants may threaten the long-term health and persistence of Cape Town's caracal population in unexpected ways, particularly when interacting with additional known pollutant and pathogen exposure. The caracal is a valuable sentinel for assessing metal exposure and can be used in pollution monitoring programmes to mitigate exposure and promote biodiversity conservation in human-dominated landscapes.

Water pollution threats in important bird and biodiversity areas from Spain.

Chemical pollution is still an underestimated threat to surface waters from natural areas. This study has analysed the presence and distribution of 59 organic micropollutants (OMPs) including pharmaceuticals, lifestyle compounds, pesticides, organophosphate esters (OPEs), benzophenone and perfluoroalkyl substances (PFASs) in 411 water samples from 140 Important Bird and Biodiversity Areas (IBAs) from Spain, to evaluate the impact of these pollutants in sites of environmental relevance. Lifestyle compounds, pharmaceuticals and OPEs were the most ubiquitous chemical families, while pesticides and PFASs showed a detection frequency below 25% of the samples. The mean concentrations detected ranged from 0.1 to 301 ng/L. According to spatial data, agricultural surface has been identified as the most important source of all OMPs in natural areas. Lifestyle compounds and PFASs have been related to the presence of artificial surface and wastewater treatment plants (WWTPs) discharges, which were also an important source of pharmaceuticals to surface waters. Fifteen out of 59 OMPs have been found at levels posing a high risk for the aquatic IBAs ecosystems, being the insecticide chlorpyrifos, the antidepressant venlafaxine and perfluorooctanesulfonic acid (PFOS) the most concerning compounds. This is the first study to quantify water pollution in IBAs and evidence that OMPs are an emerging threat to freshwater ecosystems that are essential for biodiversity conservation.

A non-invasive method to monitor farmland bird exposure to triazole fungicides.

The treatment of seeds with pesticides is an extended practice in current agriculture. There is a high risk of exposure in granivorous birds, such as the red-legged partridge (Alectoris rufa), that can consume those seeds remaining on the surface during sowing. Fungicide exposure could in turn affect bird reproductive capacity. To better understand to what extent triazole fungicides are a threat to granivorous birds, we need an easy and reliable method to quantify field exposure. In this study, we tested a novel non-invasive method to detect the presence of triazole fungicide residues in farmland bird faeces. We experimentally exposed captive red-legged partridges to validate the method, and then applied it in a real scenario to assess exposure of wild partridges. We exposed adult partridges to seeds treated with two formulations containing triazole fungicides as active ingredients: Vincit®Minima (flutriafol 2.5%) and Raxil®Plus (prothioconazole 25% and tebuconazole 15%). We collected two types of faeces (caecal and rectal samples) immediately after exposure and 7 days later and quantified the concentrations of the three triazoles and their common metabolite (1,2,4-triazole). The three active ingredients and 1,2,4-triazole were only detected in faeces collected immediately after exposure. Triazole fungicide detection rates in rectal stool were 28.6%, 73.3% and 80% for flutriafol, prothioconazole and tebuconazole, respectively. In caecal samples, detection rates were 40%, 93.3% and 33.3%, respectively. 1,2,4-triazole was detected in 53% of rectal samples. For an applied use of the method in the field, we collected 43 faecal samples from wild red-legged partridges during autumn cereal seed sowing and found detectable levels of tebuconazole in 18.6% of the analysed wild partridges. The results of the experiment were then used to estimate actual exposure levels from this prevalence value found in wild birds. Our study shows that faecal analysis can be a useful tool to assess farmland bird exposure to triazole fungicides, when samples are fresh and the method has been validated for the detection of target molecules.

Mercury-Modulated Immune Responses in Arctic Barnacle Goslings (Branta leucopsis) upon a Viral-Like Immune Challenge.

Historical mining activities in Svalbard (79°N/12°E) have caused local mercury (Hg) contamination. To address the potential immunomodulatory effects of environmental Hg on Arctic organisms, we collected newborn barnacle goslings (Branta leucopsis) and herded them in either a control or mining site, differing in Hg levels. An additional group at the mining site was exposed to extra inorganic Hg(II) via supplementary feed. Hepatic total Hg concentrations differed significantly between the control (0.011 ± 0.002 mg/kg dw), mine (0.043 ± 0.011 mg/kg dw), and supplementary feed (0.713 ± 0.137 mg/kg dw) gosling groups (average ± standard deviation). Upon immune challenge with double-stranded RNA (dsRNA) injection, endpoints for immune responses and oxidative stress were measured after 24 h. Our results indicated that Hg exposure modulated the immune responses in Arctic barnacle goslings upon a viral-like immune challenge. Increased exposure to both environmental as well as supplemental Hg reduced the level of natural antibodies, suggesting impaired humoral immunity. Hg exposure upregulated the expression of proinflammatory genes in the spleen, including inducible nitric oxide synthase (iNOS) and interleukin 18 (IL18), suggesting Hg-induced inflammatory effects. Exposure to Hg also oxidized glutathione (GSH) to glutathione disulfide (GSSG); however, goslings were capable of maintaining the redox balance by de novo synthesis of GSH. These adverse effects on the immune responses indicated that even exposure to low, environmentally relevant levels of Hg might affect immune competence at the individual level and might even increase the susceptibility of the population to infections.

Field Availability and Avoidance of Imidacloprid-Treated Soybean Seeds and Cotyledons by Birds.

Treated seeds and their cotyledons can present a toxicological risk to seed-eating birds. To assess whether avoidance behavior limits exposure and consequently the risk to birds, three fields were sown with soybeans. Half of the surface of each field was sown with seeds treated with 42 g/100 kg seed of insecticide imidacloprid (T plot, treated) and the other half with seeds without imidacloprid (C plot, control). Unburied seeds were surveyed in C and T plots at 12 and 48 h post-sowing. Damaged seedlings were surveyed in C and T plots at 12 days post-sowing. The abundance and richness of birds was surveyed at the field level (without distinguishing between C and T plots) before, during, and after sowing, and 12 days post-sowing. Unburied seed density was higher in the headlands of the T plots than in the C plots, but did not differ between 12 and 48 h. The damage to cotyledons of seedlings was 15.4% higher in C plots than in T plots. The abundance and richness/ha of birds that eat seeds and cotyledons were lower after sowing, indicating a deterrent effect on birds by sowing imidacloprid-treated seeds. Although the variation in seed density over time does not allow solid conclusions to be drawn about the avoidance of seeds treated by birds, the seedling results suggest an aversive effect of imidacloprid-treated soybeans on birds. The dominant species was the eared dove (Zenaida auriculata), whose risk of acute poisoning by imidacloprid in soybean seeds and cotyledons was low, according to its toxicity exposure ratio, foraged area of concern, and foraged time of concern. Environ Toxicol Chem 2023;42:1049-1060. © 2023 SETAC.

Legacy and emerging contaminants in flamingos' chicks' blood from the Ebro Delta Natural Park.

The Ebro Delta is a wetland of international importance for waterbird conservation but severally affected by intensive agriculture, toxic waste discharges from a past chloro-alkali industry and affluence of tourism. The discharge of contaminants associated to these activities pose waterbirds breeding in the Ebro Delta at risk. The aim of this study is to evaluate the exposure of 91 emerging and legacy micropollutants in flamingo chicks (Phoenicopterus roseus), an emblematic species of the area. Fifty chicks of 45-60 days were captured, biometric parameters measured and whole blood collected. Compounds analyzed included perfluoroalkyl substances (PFASs), pharmaceuticals, organophosphate esters (OPEs), in-use pesticides, polychlorinated biphenyls (PCBs), organochlorine pesticides (OCs), and polycyclic aromatic hydrocarbons (PAHs). The results indicate a multi-exposure of flamingo's chicks from a very young age. PFASs were the most ubiquitous compounds with ∑PFASs ranging from 9.34 to 576 ng/mL, being PFOA, PFOS and PFHxS detected in all samples. ∑PAHs ranged from 0.19 to 423 ng/mL, ∑PCBs from 0.5 to 15.6 ng/mL and ∑OCs from 1.35 to 37.8 ng/mL. Pharmaceuticals, OPEs and in-use pesticides were not detected. The flamingo's filtering behavior on mud and maternal ovo-transference are the more likely routes of exposure of organic micropollutants to flamingos' chicks. The reported levels of micropollutants were not associated with any alteration in the body condition of chicks. This is the first study to describe flamingos chicks' exposure to multiple contaminants, highlighting the importance of biomonitoring for wildlife conservation and biodiversity preservation.

A global review of the temporal and spatial patterns of DDT and dieldrin monitoring in raptors.

Concentrations of organochlorine pesticides have been extensively monitored in birds, particularly from higher trophic guilds such as raptors. While monitoring of raptors has been ongoing for decades, patterns from monitoring activities have never been summarised on a global scale. In this study, we undertake a review to better describe the monitoring of two widespread organochlorine pesticides monitored globally in raptors, DDT and dieldrin. We provide a historical retrospective on the monitoring effort of a global environmental issue. Sampling was heavily biased geographically to the global north, with more than 90 % of studies conducted in this socio-geographic region, most from Europe and North America. Although monitoring occurred from at least 114 species, most samples came from relatively few species, with three species (Eurasian Sparrowhawk Accipiter nisus, Bald Eagle Haliaeetus leucocephalus, and Peregrine Falcon Falco peregrinus) comprising 50 % of samples. The types of raptors sampled have changed over time, with avian and mammal specialists dominating samples until the 1970s, but more diverse dietary guilds monitored in later decades, and greater proportions of samples coming from generalist species. The three most sampled tissues (egg, liver, and plasma) comprised 84 % of all samples. Eggs were the earliest tissue examined and the only tissue sampled in all decades. The geographical bias in monitoring effort and relatively narrow species focus, suggests that patterns in these pesticides are unlikely to be fully representative of all global environments occupied by raptors. While DDT has been banned throughout most of the global north, it remains in use in the global south, yet monitoring effort in the south, does not match that of the north. While monitoring remains prevalent in the global north, contemporary monitoring is limited in the global south with less than 10 % of raptors sampled in Asia, Africa, and South America, over the last 3 decades.

Do Pharmaceuticals in the Environment Pose a Risk to Wildlife?

The vast majority of knowledge related to the question "To what extent do pharmaceuticals in the environment pose a risk to wildlife?" stems from the Asian vulture crisis (>99% decline of some species of Old World vultures on the Indian subcontinent related to the veterinary use of the nonsteroidal anti-inflammatory drug [NSAID] diclofenac). The hazard of diclofenac and other NSAIDs (carprofen, flunixin, ketoprofen, nimesulide, phenylbutazone) to vultures and other avian species has since been demonstrated; indeed, only meloxicam and tolfenamic acid have been found to be vulture-safe. Since diclofenac was approved for veterinary use in Spain and Italy in 2013 (home to ~95% of vultures in Europe), the risk of NSAIDs to vultures in these countries has become one of the principal concerns related to pharmaceuticals and wildlife. Many of the other bodies of work on pharmaceutical exposure, hazard and risk to wildlife also relate to adverse effects in birds (e.g., poisoning of scavenging birds in North America and Europe from animal carcasses containing pentobarbital, secondary and even tertiary poisoning of birds exposed to pesticides used in veterinary medicine as cattle dips, migratory birds as a vector for the transfer of antimicrobial and antifungal resistance). Although there is some research related to endocrine disruption in reptiles and potential exposure of aerial insectivores, there remain numerous knowledge gaps for risk posed by pharmaceuticals to amphibians, reptiles, and mammals. Developing noninvasive sampling techniques and new approach methodologies (e.g., genomic, in vitro, in silico, in ovo) is important if we are to bridge the current knowledge gaps without extensive vertebrate testing. Environ Toxicol Chem 2023;00:1-16. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Second-generation anticoagulant rodenticides in the blood of obligate and facultative European avian scavengers.

The widespread use of second-generation anticoagulant rodenticides (SGARs) and their high persistence in animal tissues has led to these compounds becoming ubiquitous in rodent-predator-scavenger food webs. Exposure to SGARs has usually been investigated in wildlife species found dead, and despite growing evidence of the potential risk of secondary poisoning of predators and scavengers, the current worldwide exposure of free-living scavenging birds to SGARs remains scarcely investigated. We present the first active monitoring of blood SGAR concentrations and prevalence in the four European obligate (i.e., vultures) and facultative (red and black kites) avian scavengers in NE Spain. We analysed 261 free-living birds and detected SGARs in 39.1% (n = 102) of individuals. Both SGAR prevalence and concentrations (ΣSGARs) were related to the age and foraging behaviour of the species studied. Black kites showed the highest prevalence (100%), followed by red kites (66.7%), Egyptian (64.2%), bearded (20.9%), griffon (16.9%) and cinereous (6.3%) vultures. Overall, both the prevalence and average ΣSGARs were higher in non-nestlings than nestlings, and in species such as kites and Egyptian vultures foraging in anthropic landscapes (e.g., landfill sites and livestock farms) and exploiting small/medium-sized carrions. Brodifacoum was most prevalent (28.8%), followed by difenacoum (16.1%), flocoumafen (12.3%) and bromadiolone (7.3%). In SGAR-positive birds, the ΣSGAR (mean ± SE) was 7.52 ± 0.95 ng mL-1; the highest level detected being 53.50 ng mL-1. The most abundant diastereomer forms were trans-bromadiolone and flocoumafen, and cis-brodifacoum and difenacoum, showing that lower impact formulations could reduce secondary exposures of non-target species. Our findings suggest that SGARs can bioaccumulate in scavenging birds, showing the potential risk to avian scavenging guilds in Europe and elsewhere. We highlight the need for further studies on the potential adverse effects associated with concentrations of SGARSs in the blood to better interpret active monitoring studies of free-living birds.

Determinants of the exposure of Eurasian griffon vultures (Gyps fulvus) to fluoroquinolones used in livestock: The role of supplementary feeding stations.

Veterinary pharmaceuticals, including antibiotics, are emerging contaminants of concern worldwide. Avian scavengers are exposed to pharmaceuticals through consumption of livestock carcasses used for feeding wildlife for conservation purposes at supplementary feeding stations. Here we tested the hypothesis that griffon vultures (Gyps fulvus) would be more exposed to antibiotics (i.e., quinolones) when feeding on livestock carcasses from intensive farming than when they rely on carcasses from extensive farming or wild animals. We sampled 657 adult griffon vultures captured between 2008 and 2012. In addition, we sampled tissues from domestic livestock supplied at feeding stations in the study area between 2009 and 2019; pig (n = 114), sheep (n = 28), cow (n = 1) and goat (n = 2). Samples were analysed by liquid chromatography with electrospray ionization mass spectrometry (LC-ESI-MS). Quinolones were detected in plasma from 12.9% of the griffon vultures analysed. Quinolone prevalence in griffon vultures varied significantly among feeding stations but was also affected by the total amount of carcasses supplemented, especially the mass of pig carcasses. These results aligned with a 21.1% quinolone prevalence in pig carcasses sampled at feeding stations, wherein enrofloxacin and ciprofloxacin levels of up to 3359 ng/g and 1550 ng/g, respectively, were found. Given enrofloxacin pharmacokinetics in pig tissues, 5.3% of the analysed pigs may have died no more than one day after treatment. Quinolone presence in vultures was negatively associated with blood lead levels, which mostly originates from lead ammunition and indicates a higher consumption of game animal carcasses. Carcass disposal for feeding avian scavengers must always assess and manage the risks posed by veterinary pharmaceuticals, especially when livestock provided may have died soon after treatment.

Factors influencing lead, mercury and other trace element exposure in birds from metal mining areas.

Non-ferrous metal mining is considered one of the largest sources of toxic metal released to the environment and may threaten ecosystems, notably biota. We explored how birds that inhabit non-ferrous metal mining sites are exposed to mercury, lead, and other trace elements by analyzing their feathers and verifying which factors may influence element concentrations in feathers. We sampled a total of 168 birds, representing 26 species, with different feeding habits and migration patterns in a non-polluted reference site and two historical metal mining areas: Almadén, which is considered one of the most heavily mercury-contaminated sites worldwide, and the Sierra Madrona mountains where lead has been mined since ancient times. The quantification of aluminum (Al), arsenic (As), barium (Ba), beryllium (Be), cadmium (Cd), cobalt (Co), chromium (Cr), copper (Cu), iron (Fe), mercury (Hg), magnesium (Mg), manganese (Mn), molybdenum (Mo), nickel (Ni), lead (Pb), selenium (Se), thorium (Th), thallium (Tl), uranium (U), vanadium (V) and zinc (Zn) was performed by inductively coupled plasma mass spectrometry (ICP-MS). Feather analysis revealed contamination by Hg and Pb, in Almadén and Sierra Madrona, respectively. We found that granivorous birds had the lowest feather Hg levels compared to those found in omnivorous, insectivorous, and piscivorous species, whereas feather Pb was about twice as high in granivores and omnivores, than in insectivorous and piscivorous birds. We also found differences among study sites in 13 elements and confirmed the influence of feather age, migratory patterns of the birds, and external deposition of elements, on metal concentrations in the feathers. Our results highlight that despite the cessation of metal mining in the study areas, local avifauna are being exposed to Hg and Pb from abandoned mines and old tailings sites, indicating that appropriate measures are needed to protect biota from overexposure to these toxic metals.

Weasel exposure to the anticoagulant rodenticide bromadiolone in agrarian landscapes of southwestern Europe.

Bromadiolone is an anticoagulant rodenticide (AR) commonly used as a plant protection product (PPP) against rodent pests in agricultural lands. ARs can be transferred trophically to predators/scavengers when they consume intoxicated live or dead rodents. ARs exposure in weasels Mustela nivalis, small mustelids specialized on rodent predation, is poorly known in southern Europe. Moreover, in this species there is no information on bioaccumulation of AR diastereomers e.g., cis- and trans-bromadiolone. Trans-bromadiolone is more persistent in the rodent liver and thus, is expected to have a greater probability of trophic transfer to predators. Here, we report on bromadiolone occurrence, total concentrations and diastereomers proportions (trans- and cis-bromadiolone) in weasels from Castilla y León (north-western Spain) collected in 2010-2017, where bromadiolone was irregularly applied to control outbreaks of common voles Microtus arvalis mainly with cereal grain bait distributed by the regional government. We also tested variables possibly associated with bromadiolone occurrence and concentration, such as individual features (e.g., sex), spatio-temporal variables (e.g., year), and exposure risk (e.g., vole outbreaks). Overall bromadiolone occurrence in weasels was 22% (n = 32, arithmetic mean of concentration of bromadiolone positives = 0.072 mg/kg). An individual showed signs of bromadiolone intoxication (i.e., evidence of macroscopic hemorrhages or hyperaemia and hepatic bromadiolone concentration > 0.1 mg/kg). All the exposed weasels (n = 7) showed only trans-bromadiolone diastereomer in liver, whilst a single analyzed bait from those applied in Castilla y León contained trans- and cis-bromadiolone at 65/35%. Bromadiolone occurrence and concentration in weasels varied yearly. Occurrence was higher in 2012 (100% of weasels), when bromadiolone was widely distributed, compared to 2016-2017 (2016: 20%; 2017: 8.33%) when bromadiolone was exceptionally permitted. The highest concentrations happened in 2014 and 2017, both years with vole outbreaks. Our findings indicate that specialist rodent predators could be exposed to bromadiolone in areas and periods with bromadiolone treatments against vole outbreaks.