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.

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.

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.

A regression-based QSAR-model to predict acute toxicity of aromatic chemicals in tadpoles of the Japanese brown frog (Rana japonica): Calibration, validation, and future developments to support risk assessment of chemicals in amphibians.

Amphibian populations are undergoing a global decline worldwide. Such decline has been attributed to their unique physiology, ecology, and exposure to multiple stressors including chemicals, temperature, and biological hazards such as fungi of the Batrachochytrium genus, viruses such as Ranavirus, and habitat reduction. There are limited toxicity data for chemicals available for amphibians and few quantitative structure-activity relationship (QSAR) models have been developed and are publicly available. Such QSARs provide important tools to assess the toxicity of chemicals particularly in a data poor context. QSARs provide important tools to assess the toxicity of chemicals particularly when no toxicological data are available. This manuscript provides a description and validation of a regression-based QSAR model to predict, in a quantitative manner, acute lethal toxicity of aromatic chemicals in tadpoles of the Japanese brown frog (Rana japonica). QSAR models for acute median lethal molar concentrations (LC50-12 h) of waterborne chemicals using the Monte Carlo method were developed. The statistical characteristics of the QSARs were described as average values obtained from five random distributions into training and validation sets. Predictions from the model gave satisfactory results for the overall training set (R2 = 0.72 and RMSE = 0.33) and were even more robust for the validation set (R2 = 0.96 and RMSE = 0.11). Further development of QSAR models in amphibians, particularly for other life stages and species, are discussed.

Assessing Stress Response in Lizards from Agroecosystems with Different Management Practices.

Despite the importance of reptiles in agroecosystems, little is known about the effects of agricultural intensification and pesticide use on these animals. We compared antioxidant and haematological biomarkers in the wild Italian wall lizards Podarcis siculus from three olive groves representing a gradient of management intensity. Lizards from the conventional grove showed induced antioxidant defences relative to those from the organic field. However, this induction did not avoid the occurrence of oxidative stress in males from intensively managed olive groves, who showed TBARS levels 58%-133% higher than males from the other sites. Haematological responses also suggested increased stress in females from the intensively managed olive groves, with a heterophil-to-lymphocyte ratio 5.3 to 14.8-fold higher than in the other sites. The observed stress responses of lizards along the studied gradient of agricultural management suggest their potential usefulness as non-destructive biomarkers to environmental stressors associated with agricultural intensification.

Bird exposure to fungicides through the consumption of treated seeds: A study of wild red-legged partridges in central Spain.

Sown seeds are a key component of many farmland birds' diets due to natural food shortages in autumn and winter. Because these seeds are often treated with pesticides, their ingestion by birds can result in toxic effects. For risk assessment, data on treated seed toxicity should be combined with information about exposure risk for wild birds and the factors that modulate it. We characterized the exposure of red-legged partridges to pesticide-treated seeds through the analysis of digestive contents of birds shot by hunters (n = 194) in an agricultural region in central Spain. We measured the contribution of sown seeds to the partridges' diet and how it related to pesticide exposure. Moreover, we evaluated the influence of landscape composition on the intake of sown seeds and pesticides by partridges. During peak sowing time, seeds constituted half (50.7%) of the fresh biomass ingested by partridges, which consumed mostly winter cereal seeds (42.3% of biomass). Residues of seven fungicides and one insecticide (active ingredients) were detected in 33.0% of birds. The presence of pesticides in digestive contents was linked to the ingestion of cereal sown seeds. Moreover, dietary exposure of birds to pesticides was modulated by landscape characteristics, being lower in areas with heterogeneous landscapes, greater habitat mosaic and more natural vegetation. The estimated dietary intake of pesticides resulting from our field observations, in combination with experimental data on pesticide toxicity, raise concerns about the risks that pesticide-treated cereal seeds pose to granivorous bird populations. Our results highlight the importance of farming landscape composition and diversification, which should be considered as a priority in the agricultural policy to mitigate pesticide risks to farmland birds through the consumption of treated seeds.

Birds feeding on tebuconazole treated seeds have reduced breeding output.

Drilled seeds are an important food resource for many farmland birds but may pose a serious risk when treated with pesticides. Most compounds currently used as seed treatment in the EU have low acute toxicity but may still affect birds in a sub-chronic or chronic way, especially considering that the sowing season lasts several weeks or months, resulting in a long exposure period for birds. Tebuconazole is a triazole fungicide widely used in agriculture but its toxicity to birds remains largely unknown. Our aim was to test if a realistic scenario of exposure to tebuconazole treated seeds affected the survival and subsequent reproduction of the red-legged partridge (Alectoris rufa). We fed captive partridges with wheat seeds treated with 0%, 20% or 100% of tebuconazole application rate during 25 days in late winter (i.e. tebuconazole dietary doses were approximately 0.2 and 1.1 mg/kg bw/day). We studied treatment effects on the physiology (i.e. body weight, biochemistry, immunology, oxidative stress, coloration) and reproduction of partridges. Exposed birds did not reduce food consumption but presented reduced plasmatic concentrations of lipids (triglycerides at both exposure doses, cholesterol at high dose) and proteins (high dose). The coloration of the eye ring was also reduced in the low dose group. Exposure ended 60 days before the first egg was laid, but still affected reproductive output: hatching rate was reduced by 23% and brood size was 1.5 times smaller in the high dose group compared with controls. No significant reproductive effects were found in the low dose group. Our results point to the need to study the potential endocrine disruption mechanism of this fungicide with lagged effects on reproduction. Risk assessments for tebuconazole use as seed treatment should be revised in light of these reported effects on bird reproduction.

Integrating active and passive monitoring to assess sublethal effects and mortality from lead poisoning in birds of prey.

The ingestion of lead (Pb) ammunition is the most important exposure pathway to this metal in birds and involve negative consequences to their health. We have performed a passive monitoring of Pb poisoning in birds of prey by measuring liver (n = 727) and blood (n = 32) Pb levels in individuals of 16 species found dead or sick in Spain between 2004 and 2020. We also performed an active monitoring by measuring blood Pb levels and biomarkers of haem biosynthesis, phosphorus (P) and calcium (Ca) metabolism, oxidative stress and immune function in individuals (n = 194) of 9 species trapped alive in the field between 2016 and 2017. Passive monitoring results revealed some species with liver Pb levels associated with severe clinical poisoning (>30 µg/g d.w. of Pb): Eurasian griffon vulture (27/257, 10.5%), red kite (1/132, 0.8%), golden eagle (4/38, 10.5%), and Northern goshawk (1/8, 12.5%). The active monitoring results showed that individuals of bearded vulture (1/3, 33.3%), Eurasian griffon vulture (87/118, 73.7%), Spanish imperial eagle (1/6, 16.7%) and red kite (1/18, 5.6%) had abnormal blood Pb levels (>20 µg/dL). Blood Pb levels increased with age, and both monitoring methods showed seasonality in Pb exposure associated with a delayed effect of the hunting season. In Eurasian griffon, blood Pb concentration was associated with lower δ-ALAD activity in blood and P levels in plasma, and with higher blood lipid peroxidation and plasma carotenoid levels in agreement with other experimental and field studies in Pb-exposed birds. The study reveals that Pb poisoning is a significant cause of death and sublethal effects on haem biosynthesis, P metabolism and oxidative stress in birds of prey in Spain.

Multi-level analysis of exposure to triazole fungicides through treated seed ingestion in the red-legged partridge.

Triazole fungicides are the most widely used products to treat cereal seeds. Granivorous birds, such as red-legged partridges (Alectoris rufa), which consume seeds left on the surface of fields after sowing, have a high risk of exposure. As triazole fungicides can affect sterol synthesis, we tested the hypothesis that treated seed consumption could alter the synthesis of sex hormones and reduce the reproductive capacity of partridges. We exposed adult partridges to seeds treated with four different formulations containing triazoles as active ingredients (flutriafol, prothioconazole, tebuconazole, and a mixture of the latter two) simulating a field exposure during the late autumn sowing season. All treatments produced biochemical changes and an overexpression of genes encoding for enzymes involved in the biosynthesis of sterols and steroid hormones, such as PMVK, ABCA1, MVD, PSCK9, DHCR7 and HSD17B7. Plasma levels of oestradiol were reduced in partridges exposed to tebuconazole. We also monitored reproduction 3 months after exposure (laying date, egg fertilization and hatching rates). We observed a 14-day delay in the laying onset of partridges that had been exposed to flutriafol as compared to controls. These results show that the consumption of seeds treated with triazole fungicides has the potential to affect granivorous bird reproduction. We recommend the evaluation of lagged reproductive effects as part of the protocols of environmental risk assessment of pesticides in wild birds in light of the effects resulting from the exposure to triazole-treated seeds.

Egg Overspray with Herbicides and Fungicides Reduces Survival of Red-Legged Partridge Chicks.

Within the environmental risk assessment conducted for pesticide registration in the European Union (EU), avian reproductive toxicity is characterized after exposing adults. However, eggs of ground-nesting species can be exposed when pesticide applications occur during laying or incubation. We simulated environmentally realistic exposure of red-legged partridge (Alectoris rufa) eggs to an herbicide (2,4-D) and a fungicide (tebuconazole) applied to winter cereal crops during the breeding season of most farmland birds. We analyzed the effects on hatching success, offspring survival, and physiology. Exposure by overspray led to greater pesticide accumulation in the eggshell or content than exposure through contact with treated soil (3.1-13.7 times higher, depending on the pesticide and target sample). Egg overspray with tebuconazole significantly increased chick mortality, which was 26% higher than that of controls. 2,4-D caused a similar but a close to significant increase (chick mortality 24% higher than controls). Exposure to either pesticide through contact with treated soils did not affect chick survival but altered some biochemical parameters posthatching. Our experiment shows that egg spraying with pesticides should be considered as a relevant exposure scenario in risk assessment procedures, given its potential to affect the reproductive success of ground-nesting farmland birds.

Assessment of ecotoxicological risks to river otters from ingestion of invasive red swamp crayfish in metal contaminated areas: Use of feces to estimate dietary exposure.

The invasive red swamp crayfish (Procambarus clarkii) has become a major food resource for Eurasian otters (Lutra lutra) in the Iberian Peninsula. Crayfish accumulate large amounts of metals, and hence otters could be at risk of exposure and intoxication through crayfish consumption. We conducted a food safety risk assessment for otters inhabiting two historical mining areas in central Spain affected by lead (Pb) and mercury (Hg) pollution. Estimated daily intakes (EDI) of Pb and Hg were non-invasively calculated from the proportion of crayfish remains and metal levels in otter feces. We considered that the abdominal muscle and the carcass of crayfish differ significantly in relative weight, total metal content and bioavailability of metals to reduce the uncertainty of risk characterization. Fecal concentrations of Hg and Pb in the polluted areas were 1.878 and 6.554 µg/g d. w., respectively (13-fold and 7-fold higher compared to a non-polluted area). EDI of Hg and Pb in the polluted areas were 66.02 and 78.26 µg/kg-day, respectively (14- and 8-fold higher than in the reference area). EDI from the Hg area were above minimum levels susceptible to cause neurotoxicity in mustelids, and 6.3% were above levels susceptible to cause histopathological lessions. In the Pb area, 16.7% of EDI were consistent with levels causing reproductive effects. Metal exposure through crayfish consumption might prevent or slow the recovey of otters in these polluted environments, thus this factor should be considered in management strategies aimed to protect otter populations.

Food safety risk assessment of metal pollution in crayfish from two historical mining areas: Accounting for bioavailability and cooking extractability.

Here we characterize the bioaccumulation of mercury (Hg) and lead (Pb) in red swamp crayfish (Procambarus clarkii) from two river courses in Central Spain that are impacted by historical Hg and Pb mining activities, respectively. We estimate the absolute oral bioavailability of metals in crayfish tissues by means of in vitro bioaccessibility simulations, and assess whether their consumption may imply a health risk for humans by estimating target hazard quotients and safe consumption rates. We also study the effect of cooking crayfish on the mobilization of the metal body burden in the context of the traditional Spanish cuisine. The results showed that crayfish from the mining districts accumulated a high level of Hg and Pb pollution in both the tail muscle and the carcass. The in vitro bioaccessibility of Hg and Pb in the edible part was 27.86 ±â€¯4.05 and 33.73 ±â€¯5.91%, respectively. Absolute bioavailability was estimated to be 38.31 for Hg, and 20.21 (adults) and 67.35% (children) for Pb. Risk indices indicated that, even after adjusting for bioavailability, it is not safe to consume crayfish from the mining-impacted rivers because of their high levels of Hg and Pb. Using the carcass as a condiment for flavouring should also be avoided. The cooking procedure extracted relatively small amounts of the total Hg (8.92 ±â€¯2.13%) and Pb (1.68 ±â€¯0.29%) body burden. Further research that will support human and ecological risk assessment, along with the implementation of advisory measures for the local population as regards crayfish consumption, are recommended.

Accumulation of pollutants in nestlings of an endangered avian scavenger related to territory urbanization and physiological biomarkers.

We monitor pollutant accumulation and investigate associated changes at the physiological level within the population of an obligate avian scavenger, the Egyptian Vulture (Neophron percnopterus), from Catalonia (NE Spain). This population is expanding its range, presumably because of the use of human waste disposal sites as food resource. We hypothesized that habitat urbanization, presumably associated with feeding from human wastes, could influence the accumulation of persistent organic pollutants and metals. The aim of this study was to explore the relationship between accumulated pollutants and biochemical parameters in nestling blood. We used the proportion of urban surface within an 8 km radius of each nest as a proxy to study the relationship between anthropic influence and pollutant accumulation. Observed blood levels of metals, organochlorine pesticides, polychlorinated biphenyls (PCBs), per- and polyfluoroalkylated substances (PFAS) and polybrominated diphenyl ethers (PBDEs) were relatively low, as expected for nestling individuals due to short-term exposures. CB-180 and PBDEs were associated with variations in blood biochemistry parameters; hexa-BDEs appeared positively associated with activities of the enzymes aspartate aminotransferase and lactate dehydrogenase, whereas CB-180 accumulation was associated with an increased activity of creatine phosphokinase and elevated glutathione levels. Increased CB-180 levels were also related to decreased blood concentrations of calcium, cholesterol, α-tocopherol and lutein. A proportion of urban surfaces as low as 6.56% within a radius of 8 km around the nest appears related to the accumulation of CB-180, the majority of analysed PFAS and of PBDE congeners 99 and 209, and increased urbanization was also associated with decreased plasma levels of α-tocopherol and carotenoids. These associations suggest that changes in blood profiles of vitamins, carotenoids or other analytes, despite related to increased plasma levels of CB-180, would be consequence of exploitation of artificial food sources rather than of a direct effect of the pollutants.

Immunotoxic effects of lead on birds.

The ingestion of lead (Pb) shot pellets is a well-known cause of avian mortality, but exposure of birds to Pb may have other sublethal effects that can affect population sustainability. One of these effects is the alteration of the immunological status. Pb can affect most components of the avian immune system and imbalance the relationship among them. Pb exposure typically alters the ratio between Th1- and Th2-type responses mounted by different classes of T-lymphocytes, causing the depression of the Th1 responses that are associated with cell-mediated immunity. Immunodepressing effects of Pb on birds are observed at blood levels above 50 µg dL-1, but developing birds show immunodepressing effects at much lower concentrations (> 10 µg dL-1). Impacts of Pb on the avian immune system also relate to reduced resistance to infection. We review immunotoxic effects of Pb on birds affected by shot ingestion as well as by other sources of exposure to this element.

Tools for non-invasive sampling of metal accumulation and its effects in Mediterranean pond turtle populations inhabiting mining areas.

Among reptiles, freshwater turtle species have high potential for metal accumulation because of their long lifespan or their aquatic and terrestrial habits. In order to monitor metal bioaccumulation, determine potential toxic effects, and investigate tools for non-invasive metal sampling in reptiles, we studied lead (Pb) and mercury (Hg) accumulation in Mediterranean pond turtles (Mauremys leprosa) inhabiting two former mining areas, one of them with high environmental concentrations of Pb (Sierra Madrona-Alcudia Valley district) and the other one with high environmental concentrations of Hg (Almadén district). Individuals from the Pb mining area showed mean blood concentrations (i.e. 5.59 µg Pb/g dry weight, d.w.) that were higher than those measured in other populations. Blood Hg concentrations were highest (8.83 µg Hg/g d.w.) in the site close to the former Hg mines, whereas blood Hg concentrations in terrapins from another site of Almadén district, located ∼28 km downstream, were not different from locations at the non-mining area. Animals from the Pb-contaminated site showed evidence of oxidative stress, whereas those from the Hg-contaminated site showed increased activity of the antioxidant enzyme glutathione peroxidase, as well as reduced circulating levels of the main endogenous antioxidant peptide, glutathione. Concentrations measured in feces and carapace scutes were useful indicators to monitor blood concentrations of Pb, but not of Hg. Our results provide evidence of the usefulness of freshwater turtles as sentinels of chronic metal pollution, and validate non-invasive tools to advance Pb monitoring in reptiles.

Domestic waste disposal sites secure food availability but diminish plasma antioxidants in Egyptian vulture.

Some domestic waste disposal sites (DWDS) accumulate enormous amounts of food that humans discard. This food resource is exploited by different species whose population dynamics have as a result been altered. The effects of feeding on human waste on individual health parameters such as body condition are still unknown. Traditionally, body condition in wildlife has been assessed using morphological indices based on size and weight. Expanding the range of indices to include the analysis of indicators at the physiological level can provide a more comprehensive picture of the animals' health status. In this work, we studied how different amounts of waste consumption (assessed using stable isotope analysis) affect nestling health and body condition in an expanding population of the endangered Egyptian Vulture (Neophron percnopterus). We used morphometric and physiological approaches, including plasma biochemistry, antioxidant defences and oxidative stress biomarkers. The results of the multivariate redundancy analysis and variance partitioning showed that the proportion of diet originating from DWDS is the main factor explaining physiological indicators in the studied vulture population. Nestlings that were fed on food from DWDS had lower levels of vitamins (e.g. α-tocopherol) and carotenoids (zeaxanthin and lutein), which have important functions as antioxidant defences, immunostimulant properties and ornamental purposes. Nevertheless, nestlings that were fed on food from DWDS were better nourished and experienced fewer food shortages than individuals not fed using this resource, which had lower levels of cholesterol. Possibly due to the easy access to food, breeding pairs using DWDS could have fewer difficulties in feeding their nestlings and thus could engender more regular brood sizes than those not exploiting this type of resource. Our integrative approach using diverse components was crucial for a reliable assessment of health and body condition in Egyptian Vulture nestlings and its relationship to the consumption of food derived from human-altered environments.

Exposure of black-necked grebes (Podiceps nigricollis) to metal pollution during the moulting period in the Odiel Marshes, Southwest Spain.

European populations of black-necked grebes (Podiceps nigricollis) congregate every year to moult at the salt ponds of the Odiel Marshes (SW Spain). However, the Odiel Marshes are part of one of the most metal-polluted coastal estuaries in the world, which may pose risks to wildlife. We assessed the exposure of grebes to metal pollution during the critical moulting period in the Odiel Marshes and its potential to cause adverse health effects. Levels of metals in red blood pellet (as a biomarker of exposure), plasma carotenoids, eye redness, and body condition (as biomarkers of effects) were studied. Metal content was also analyzed in the brine shrimp Artemia parthenogenetica, the most important food for grebes in this hypersaline ecosystem during the moulting period. Results showed that, in comparison to toxicity thresholds, grebes had relatively high blood levels of arsenic (As), mercury (Hg) and zinc (Zn). The high loads found in Artemia and the way blood levels vary during the moulting period indicate that shrimp consumption may be the main route of metal exposure for grebes. Plasma carotenoids and body condition showed a positive association with exposure to As, while the relationship of lutein-like carotenoids with Hg accumulation was negative at the beginning of the moulting period to become positive afterwards. Moreover, eye redness was negatively affected by As accumulation. Factors including food resource availability, seasonal fluctuations in physiological status, and interannual variations in the degree of environmental contamination should be considered in monitoring efforts when using moult migrant waterbirds as sentinel species.

Brood size is reduced by half in birds feeding on flutriafol-treated seeds below the recommended application rate.

Despite the efforts of the European Commission to implement measures that offset the detrimental effects of agricultural intensification, farmland bird populations continue to decline. Pesticide use has been pointed out as a major cause of decline, with growing concern about those agro-chemicals that act as endocrine disruptors. We report here on the effects of flutriafol, a ubiquitous systemic fungicide used for cereal seed treatment, on the physiology and reproduction of a declining gamebird. Captive red-legged partridges (Alectoris rufa; n = 11-13 pairs per treatment) were fed wheat treated with 0%, 20% or 100% of the flutriafol application rate during 25 days in late winter. We studied treatment effects on the reproductive performance, carotenoid-based coloration and cellular immune responsiveness of adult partridges, and their relationship with changes in oxidative stress biomarkers and plasma biochemistry. We also studied the effect of parental exposure on egg antioxidant content and on the survival, growth and cellular immune response of offspring. Exposed partridges experienced physiological effects (reduced levels of cholesterol and triglycerides), phenotypical effects (a reduction in the carotenoid-based pigmentation of their eye rings), and most importantly, severe adverse effects on reproduction: a reduced clutch size and fertile egg ratio, and an overall offspring production reduced by more than 50%. No effects on body condition or cellular immune response of either exposed adult or their surviving offspring were observed. These results, together with previous data on field exposure in wild partridges, demonstrate that seed treatment with flutriafol represents a risk for granivorous birds; they also highlight a need to improve the current regulation system used for foreseeing and preventing negative impacts of Plant Protection Products on wildlife.

Toward sustainable environmental quality: Priority research questions for Europe.

The United Nations' Sustainable Development Goals have been established to end poverty, protect the planet, and ensure prosperity for all. Delivery of the Sustainable Development Goals will require a healthy and productive environment. An understanding of the impacts of chemicals which can negatively impact environmental health is therefore essential to the delivery of the Sustainable Development Goals. However, current research on and regulation of chemicals in the environment tend to take a simplistic view and do not account for the complexity of the real world, which inhibits the way we manage chemicals. There is therefore an urgent need for a step change in the way we study and communicate the impacts and control of chemicals in the natural environment. To do this requires the major research questions to be identified so that resources are focused on questions that really matter. We present the findings of a horizon-scanning exercise to identify research priorities of the European environmental science community around chemicals in the environment. Using the key questions approach, we identified 22 questions of priority. These questions covered overarching questions about which chemicals we should be most concerned about and where, impacts of global megatrends, protection goals, and sustainability of chemicals; the development and parameterization of assessment and management frameworks; and mechanisms to maximize the impact of the research. The research questions identified provide a first-step in the path forward for the research, regulatory, and business communities to better assess and manage chemicals in the natural environment. Environ Toxicol Chem 2018;37:2281-2295. © 2018 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

Validity of fish, birds and mammals as surrogates for amphibians and reptiles in pesticide toxicity assessment.

Amphibians and reptiles are the two most endangered groups of vertebrates. Environmental pollution by pesticides is recognised as one of the major factors threatening populations of these groups. However, the effects of pesticides on amphibians and reptiles have been studied for few substances, which is partly related to the fact that these animals are not included in the mandatory toxicity testing conducted as part of environmental risk assessments of pesticides. Whether risks of pesticides to amphibians and reptiles are addressed by surrogate taxa used in risk assessment is currently under debate. In order to develop a scientifically sound and robust risk assessment scheme, information needs to be gathered to examine whether fish, birds and mammals are valid surrogates for amphibians and reptiles. We updated a systematic review of scientific literature that was recently published compiling toxicity data on amphibians and reptiles. The outcome of this review was analysed with the purposes to (1) compare endpoints from amphibians and reptiles with the available information from fish, birds and mammals, and (2) develop species sensitivity distributions (SSDs) for those substances tested in at least six amphibian species (no substances were found tested in at least six reptile species) to identify a candidate amphibian model species to be used as surrogate in risk assessment. A positive correlation was found between toxicity recorded on fish and amphibians, the former revealing, in general, to be more sensitive than the latter to waterborne pollutants. In the terrestrial environment, although birds and mammals were more sensitive than amphibians and reptiles to at least 60% of tested substances, just a few weak significant correlations were observed. As a general rule, homoeothermic vertebrates are not good surrogates for reptiles and terrestrial amphibians in pesticide risk assessment. However, some chemical-dependent trends were detected, with pyrethroids and organochlorine insecticides being more toxic to amphibians or reptiles than to birds or mammals. These trends could ultimately help in decisions about protection provided by surrogate taxa for specific groups of substances, and also to determine when risk assessment of pesticides needs to pay special consideration to amphibians and reptiles. The outcome of this review reflects that there is still much information needed to reduce uncertainties and extract relevant conclusions on the overall protection of amphibians and reptiles by surrogate vertebrates.