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.

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.

Neonicotinoid insecticides can be transported from agricultural fields, where they are used as foliar sprays or seed treatments, to surface waters by surface or sub-surface runoff. Few studies have investigated the toxicity of neonicotinoid or the related butenolide insecticides to freshwater mollusk species. The current study examined the effect of neonicotinoid and butenolide exposures to the early-life stages of the ramshorn snail, Planorbella pilsbryi, and the wavy-rayed lampmussel, Lampsilis fasciola. Juvenile P. pilsbryi were exposed to imidacloprid, clothianidin, or thiamethoxam for 7 or 28 d and mortality, growth, and biomass production were measured. The viability of larval (glochidia) L. fasciola was monitored during a 48 h exposure to six neonicotinoids (imidacloprid, thiamethoxam, clothianidin, acetamiprid, thiacloprid, or dinotefuran), or a butenolide (flupyradifurone). The 7-d LC50s of P. pilsbryi for imidacloprid, clothianidin, and thiamethoxam were ≥4000 μg/L and the 28-d LC50s were ≥182 μg/L. Growth and biomass production were considerably more sensitive endpoints than mortality with EC50s ranging from 33.2 to 122.0 μg/L. The 48-h LC50s for the viability of glochidia were ≥456 μg/L for all seven insecticides tested. Our data indicate that neonicotinoid and butenolide insecticides pose less of a hazard with respect to mortality of the two species of mollusk compared to the potential hazard to other non-target aquatic insects.

In the present study we analyze the effect of seed treatment by a range of nano-TiO2 concentrations on the growth of Arabidopsis thaliana plants, on the vitamin E content and the expression of its biosynthetic genes, as well as activity of antioxidant enzymes and lipid peroxidation. To conduct the mechanistic analysis of nano-TiO2 on plants growth and antioxidant status we applied nanoparticles concentrations that are much higher than those reported in the environment. We find that as the concentration of nano-TiO2 increases, the biomass, and chlorophyll content in 5-week-old Arabidopsis thaliana plants decrease in a concentration dependent manner. In opposite, higher nano-TiO2 concentration enhanced root growth. Our results indicate that a high concentration of nano-TiO2 induces symptoms of toxicity and elevates the antioxidant level. We also find that the expression levels of tocopherol biosynthetic genes were either down- or upregulated in response to nano-TiO2. Thermoluminescence analysis shows that higher nano-TiO2 concentrations cause lipid peroxidation. To the best of our knowledge, this is the first report concerning the effect of nano-TiO2 on vitamin E status in plants. We conclude that nano-TiO2 affects the antioxidant response in Arabidopsis thaliana plants. This could be an effect of a changes in vitamin E gene expression that is diminished under lower tested nano-TiO2 concentrations and elevated under 1000 μg/ml.

Neonicotinoid insecticides are of environmental concern, but little is known about their occurrence in surface water. An area of intense corn and soybean production in the Midwestern United States was chosen to study this issue because of the high agricultural use of neonicotinoids via both seed treatments and other forms of application. Water samples were collected from nine stream sites during the 2013 growing season. The results for the 79 water samples documented similar patterns among sites for both frequency of detection and concentration (maximum:median) with clothianidin (75%, 257 ng/L:8.2 ng/L) > thiamethoxam (47%, 185 ng/L:<2 ng/L) > imidacloprid (23%, 42.7 ng/L: <2 ng/L). Neonicotinoids were detected at all nine sites sampled even though the basin areas spanned four orders of magnitude. Temporal patterns in concentrations reveal pulses of neonicotinoids associated with rainfall events during crop planting, suggesting seed treatments as their likely source.

International audience | Since their discovery in the late 1980s, neonicotinoid pesticides have become the most widely used class of insecticides worldwide, with large-scale applications ranging from plant protection (crops, vegetables, fruits), veterinary products, and biocides to invertebrate pest control in fish farming. In this review, we address the phenyl-pyrazole fipronil together with neonicotinoids because of similarities in their toxicity, physicochemical profiles, and presence in the environment. Neonicotinoids and fipronil currently account for approximately one third of the world insecticide market; the annual world production of the archetype neonicotinoid, imidacloprid, was estimated to be ca. 20,000 tonnes active substance in 2010. There were several reasons for the initial success of neonicotinoids and fipronil: (1) there was no known pesticide resistance in target pests, mainly because of their recent development, (2) their physicochemical properties included many advantages over previous generations of insecticides (i.e., organophosphates, carbamates, pyrethroids, etc.), and (3) they shared an assumed reduced operator and consumer risk. Due to their systemic nature, they are taken up by the roots or leaves and translocated to all parts of the plant, which, in turn, makes them effectively toxic to herbivorous insects. The toxicity persists for a variable period of time—depending on the plant, its growth stage, and the amount of pesticide applied. Awide variety of applications are available, including the most common prophylactic non-Good Agricultural Practices (GAP) application by seed coating. As a result of their extensive use and physicochemical properties, these substances can be found in all environmental compartments including soil, water, and air. Neonicotinoids and fipronil operate by disrupting neural transmission in the central nervous system of invertebrates. Neonicotinoids mimic the action of neurotransmitters, while fipronil inhibits neuronal receptors. In doing so, they continuously stimulate neurons leading ultimately to death of target invertebrates. Like virtually all insecticides, they can also have lethal and sublethal impacts on non-target organisms, including insect predators and vertebrates. Furthermore, a range of synergistic effects with other stressors have been documented. Here, we review extensively their metabolic pathways, showing how they form both compound-specific and common metabolites which can themselves be toxic. These may result in prolonged toxicity. Considering their wide commercial expansion, mode of action, the systemic properties in plants, persistence and environmental fate, coupled with limited information about the toxicity profiles of these compounds and their metabolites, neonicotinoids and fipronil may entail significant risks to the environment. A global evaluation of the potential collateral effects of their use is therefore timely. The present paper and subsequent chapters in this review of the global literature explore these risks and show a growing body of evidence that persistent, low concentrations of these insecticides pose serious risks of undesirable environmental impacts.

Aim of this study was to evaluate the interactive effects of glycine, alanine, calcium nitrate [Ca(NO₃)₂], and their mixture on the growth of two wheat (Triticum aestivum L.) varieties, i.e., var. Punjab-2011 and var. Anaj-2017 under lead [0.5 mM Pb(NO₃)₂] stress. A pot experiment was conducted for this purpose. Pre-sowing seed treatment with 1 mM glycine, alanine, and calcium nitrate [Ca(NO₃)₂] was applied under two levels of lead nitrate [Pb(NO₃)₂] stress, i.e., control and 0.5 mM Pb(NO₃)₂. Lead (0.5 mM) stress significantly decreased root and shoot lengths, fresh and dry weights of root and shoot, and chlorophyll contents, while it increased activities of antioxidant enzymes such as catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), and peroxidase (POD) in both wheat varieties. Lead (0.5 mM) stress increased the accumulation of free proline, glycinebetaine, total free amino acids, and total soluble protein contents. Although var. Punjab-2011 was higher in root fresh and dry weights, shoot length, and total leaf area per plant, however, var. Anaj-2017 showed less reduction in shoot dry weight, root fresh weight, and shoot length under lead stress. Under lead stress, Punjab-2011 was higher in grain yield and number of grain plant⁻¹, chlorophyll a contents, membrane permeability (%), POD activity, total free amino acids, and glycinebetaine (GB) contents as compared to Anaj-2017. Pre-sowing seed treatments with glycine, alanine, calcium nitrate, and their mixture (1 mM of each) increased shoot dry weight, number of grains per plants, 100-grain weight, number of spikes, and chlorophyll a contents under normal and lead-stressed conditions. Wheat var. Anaj-2017 showed higher growth and yield attributes as compared to var. Punjab-2011. Results of the current study have shown that pre-sowing seed treatments with glycine, alanine, calcium nitrate, and their mixture (1 mM of each) can overcome the harmful effects of lead (Pb) stress in wheat plants.

Fungicidal activity of experimental tebuconazole (TEB) formulations was investigated in laboratory soil ecosystems in wheat plant communities infected by Fusarium moniliforme. TEB was embedded in the matrix of poly-3-hydroxybutyrate, shaped as films and microgranules. These formulations were buried in the soil with wheat plants, and their efficacy was compared with that of commercial formulation Raxil and with the effect of pre-sowing treatment of seeds. In the experiment with the initially infected seeds and a relatively low level of natural soil infection caused by Fusarium fungi, the effects of the experimental P(3HB)/TEB formulations and Raxil were comparable. However, when the level of soil infection was increased by adding F. moniliforme spores, P(3HB)/TEB granules and films reduced the total counts of fungi and the abundance of F. moniliforme more effectively than Raxil. Seed treatment or soil treatment with Raxil solution showed an increase in the percentage of rot-damaged roots in the later stages of the experiment. In the early stage (between days 10 and 20), the percentage of rot-damaged roots in the soil with TEB embedded in the slowly degraded P(3HB) matrix was similar to that in the soil with Raxil. However, the efficacy of P(3HB)/TEB formulations lasted longer, and in later stages (between days 20 and 30), the percentage of rot-damaged roots in that group did not grow. In experiments with different TEB formulations and, hence, different fungicidal activities, the increase in plant biomass was 15–17 to 40–60% higher than in the groups where TEB was applied by using conventional techniques.

Systemic insecticides are applied to plants using a wide variety of methods, ranging from foliar sprays to seed treatments and soil drenches. Neonicotinoids and fipronil are among the most widely used pesticides in the world. Their popularity is largely due to their high toxicity to invertebrates, the ease and flexibility with which they can be applied, their long persistence, and their systemic nature, which ensures that they spread to all parts of the target crop. However, these properties also increase the probability of environmental contamination and exposure of nontarget organisms. Environmental contamination occurs via a number of routes including dust generated during drilling of dressed seeds, contamination and accumulation in arable soils and soil water, runoff into waterways, and uptake of pesticides by nontarget plants via their roots or dust deposition on leaves. Persistence in soils, waterways, and nontarget plants is variable but can be prolonged; for example, the half-lives of neonicotinoids in soils can exceed 1,000 days, so they can accumulate when used repeatedly. Similarly, they can persist in woody plants for periods exceeding 1 year. Breakdown results in toxic metabolites, though concentrations of these in the environment are rarely measured. Overall, there is strong evidence that soils, waterways, and plants in agricultural environments and neighboring areas are contaminated with variable levels of neonicotinoids or fipronil mixtures and their metabolites (soil, parts per billion (ppb)-parts per million (ppm) range; water, parts per trillion (ppt)-ppb range; and plants, ppb-ppm range). This provides multiple routes for chronic (and acute in some cases) exposure of nontarget animals. For example, pollinators are exposed through direct contact with dust during drilling; consumption of pollen, nectar, or guttation drops from seed-treated crops, water, and consumption of contaminated pollen and nectar from wild flowers and trees growing near-treated crops. Studies of food stores in honeybee colonies from across the globe demonstrate that colonies are routinely and chronically exposed to neonicotinoids, fipronil, and their metabolites (generally in the 1–100 ppb range), mixed with other pesticides some of which are known to act synergistically with neonicotinoids. Other nontarget organisms, particularly those inhabiting soils, aquatic habitats, or herbivorous insects feeding on noncrop plants in farmland, will also inevitably receive exposure, although data are generally lacking for these groups. We summarize the current state of knowledge regarding the environmental fate of these compounds by outlining what is known about the chemical properties of these compounds, and placing these properties in the context of modern agricultural practices.

BACKGROUND, AIM AND SCOPE: Seed treatments are widely used on cereals and other annual crops throughout Europe. Most of the formulated pesticide is found on the outside of the seed, the husk. Risk assessments of seed treatments are especially needed for granivorous mice living in the agricultural landscape e.g. for registration using the guidance for risk assessment for birds and mammals (EFSA 2009). The dehusking of seeds before consumption is a known behaviour of these mammals, but so far, no quantitative data on the reduction of exposure of seed treatments by dehusking were published. Therefore, we aimed at providing a first quantitative estimate of this behaviour-related exposure reduction for the wood mouse (Apodemus sylvaticus) with different seed types. MATERIALS AND METHODS: We evaluated the efficiency of dehusking behaviour of 20 wood mice captured in the wild for four different seeds (wheat, barley, maize and sunflower). One experimental setup used a fungicide seed treatment where the remaining seed husks of consumed seeds were analysed with a HPLC-MS/MS technique. In the second setup, we measured generic pigment present in a blank seed treatment formulation and determined the leftover pigment in the husks with a photometric technique. RESULTS: The exposure reduction was similar for the fungicide and the pigment design where the same seed types were studied. We could demonstrate exposure reductions ranging from around 60% for cereals to almost 100% for sunflower seeds as a result of the dehusking behaviour. DISCUSSION: Since exposure reduction was similar in both approaches, working with pigments would be a generic way to estimate the impact of dehusking behaviour on seed treatment exposure. This behaviour can result in a substantial exposure reduction and should, therefore, be considered in a seed-type specific way in the risk assessment of pesticide seed treatments. CONCLUSIONS: It is proposed to include a seed-specific dehusking factor in the calculations of estimated theoretical exposure of seed treatments for granivorous mice. The approach of accounting for a dehusking-related exposure reduction by field relevant wild mammal species seems a more promising way to advance the risk assessment instead of using generic species and neglecting behavioural traits. The pigment approach could be used to gather data for exposure reduction for other species and seed types. Its advantage is that it is harmless to the test species and comparatively cheap since no chemical analysis is involved. RECOMMENDATIONS AND PERSPECTIVES: Seed treatments are used for most of the cereal crops grown in Europe today. Their advantages usually include a lower application rate and the reduction of drift compared to a conventional spraying regime. However, there is a potential risk especially for granivorous mice, and its assessment is challenging in case of a high residue concentration on the dressed seeds. The concept of a dehusking factor in the risk assessment scheme for seed treatments for granivorous mice is a valid approach to account for the behavioural exposure reduction, and generic data could be easily generated also for other wild mammal species and other seed types, possibly analysing the pigment in commercial seed treatment formulations.