Background Wild pollinator populations in the European Union lack defined, quantitative Specific Protection Goals (SPGs) for pesticide risk assessment. Without an agreed threshold, lower-tier toxicity data cannot be evaluated for acceptability, nor can higher-tier study results be assessed against a protection goal, leaving the tiered ERA process at an impasse. The existing honey bee surrogate compounds the problem: this was selected for laboratory convenience rather than functional representativeness, and their managed, colonial life history cannot reliably predict outcomes for the diverse life-history strategies of wild pollinators. Results This paper presents a pragmatic, staged framework combining trait-based species selection with demographic toxicity assessment. It focuses on Stage 1 (non-spatial) of a the three-stage process, where life-stage-structured population models are implementable now (2026–2027), uses existing data and tools, and generates quantitative, population-level SPG thresholds within the current [1] tiered assessment framework. The framework combines two methodological advances: trait-based species selection identifies functionally vulnerable species by grouping taxa according to exposure- and sensitivity-related traits and population resilience; and demographic toxicity assessment translates individual-level pesticide effects into population-level outcomes through life-stage-structured projection matrices. Parameter space exploration demonstrates that Stage 1 yields defined, quantitative thresholds: a maximum acceptable population decline and a maximum acceptable exceedance risk probability. Conclusions Stage 1 resolves the immediate operational constraint by providing provisional, population-level thresholds for regulatory use. A roadmap for adaptive implementation and progressive refinement is provided: Stage 2 (operational 2028–2029) incorporates landscape-scale spatial dynamics, and Stage 3 (operational 2030+) implements full agent-based modelling using the ALMaSS system. The approach is demonstrated for bees (bumble bees and solitary bees), but the trait-based framework is designed to be flexible enough to generalise across hoverflies, butterflies, moths, and other pollinating taxa.

Insecticides are widely used to control pests, but can also be toxic to ecosystem service providers, including bees. Moreover, their efficacy can increase if they are applied in mixtures. These mixtures can be intentional, e.g., two or more active substances formulated in one product or created by farmers as tank mixtures (i.e., a mixture of different, individually formulated plant protection products (PPP) applied together in one application event or applied at short intervals), or unintentional, when exposure to a mixture results from bees foraging on different crops, each of which is treated with a different PPP. Although studies on the combined toxicity of various PPPs are increasing, the impact of mixture is still not adequately addressed in environmental risk assessment, posing a potential threat to bees. Here, we assessed the interactive effects of five insecticides belonging to different chemical classes: organophosphate Dursban 480 EC (a.s. chlorpyrifos), two pyrethroids - Sherpa 100 EC (a.s. cypermethrin) and Karate Zeon 050 CS (a.s. lambda-cyhalothrin), neonicotinoid Mospilan 20 SP (a.s. acetamiprid), and sulfoximine Closer (a.s. sulfoxaflor)) applied topically alone and as binary mixtures, on survival of Osmia bicornis females. Contrary to expectations, the experiment revealed either no interaction (in 'Dursban × Sherpa' experiment) or antagonistic interactions, particularly in mixtures of insecticide belonging to pyrethroids (Sherpa or Karate) with the one belonging to either neonicotinoids (Mospilan) or sulfoximines (Closer). Moreover, the mixture 'Karate × Closer' suggests a potential antagonistic effect on the survival of O. bicornis already at field-relevant concentrations. Such unexpected results emphasize the need for longer-term testing of cumulative toxicity effects via different exposure routes and on different bee species, as well as the need to consider the risk of exposure to multiple pesticides when assessing the safety of pesticides for bees.

Many carabid species are essential for pest management in agricultural areas, but at the same time can be exposed to pesticides. To understand how pesticides may affect ecosystem services provided by carabids, it is crucial to assess the effects of exposure not only for individual species but for whole communities. The objective of our study was thus to assess the distribution of sensitivity in carabid communities inhabiting agricultural landscapes toward three insecticides representing three major groups: neonicotinoid acetamiprid, organophosphate chlorpyrifos, and pyrethroid λ-cyhalothrin. Using a vial test, in which an active ingredient dissolved in acetone is evenly distributed inside a glass vial, we first assessed the median lethal doses (24-h LD50) for each insecticide to 10-20 species collected in agricultural areas, which were further used to establish Species Sensitivity Distribution (SSD) profiles. Each insecticide tested caused a serious threat to beetle communities at the recommended field doses, with the strongest effect caused by chlorpyrifos and the weakest by acetamiprid. The estimated Potentially Affected Fraction (PAF) of species at doses of the insecticides recommended for field use was 100 % for chlorpyrifos, 99.8 % for λ-cyhalothrin, and ca. 66 % for acetamiprid. The Hazardous Doses for 5 % species (HD5) were 0.034, 0.47, and 3.79 % of their recommended field doses, respectively. Our findings prove that carabids are a group of ecosystem service providers at considerable risk from insecticide exposure.

Bats, as exceptionally long-lived small mammals, are at particular risk of metal poisoning due to the tendency of metals to bioaccumulate throughout their lives. In our study, we investigated the general question of how trace metal concentrations change with age in different bat tissues on the example of Pipistrellus kuhlii lepidus, which lives for years in one area and is strongly associated with urban environments. To determine the exact age of the individuals, osteochronology was applied, counting the number of dentine rings in cross-sections of the upper canine tooth of each individual. The age of 57 individuals of P. kuhlii lepidus, representing ca. 10

The trait-based approach is increasingly used for soil invertebrates. Complementary to the taxonomy based approach, the trait-based approach can provide a more mechanistic understanding of the responses of organisms to environmental disturbances and of their effects on soil functioning. However, this approach has several limitations linked to the conceptual development of functional traits. There is a large ambiguity and variability in using the term "functional traits” by zoologists and ecologists working with soil invertebrates. In this study, we used a questionnaire and literature scanning to review the practical use of functional traits concept in soil ecology over the last decade. We clarifed and expanded the functional trait definition as "A functional trait is a measurable characteristic of an individual organism or its colony that has a clear link to the organism's fitness and/or its effect on other organisms and/or the environment”. We also reviewed existing trait databases showing a high amount, but also high heterogeneity and low accessibility of data on the functional traits of soil invertebrates. We suggest synthesising existing trait data and databases, using the functional trait-based approach consistently and reproducibly, and disseminating it to facilitate mechanistic research in soil ecology.

The use of pesticides has increased considerably over the past few decades to maximize crop yields. This resulted in the presence of a range of toxic compounds in ecosystems, which may interact with each other, potentially enhancing adverse impacts on wildlife and ecosystem services. The objective of this study was to evaluate potential effects induced by chronic exposure to a mixture of pesticide formulations extensively used in agriculture: one containing cypermethrin (Sherpa 100 EC - SHP) and used as insecticide, the other containing tebuconazole (Spekfree 430 SC - SKF) and used as fungicide. In laboratory experiments, the earthworm Eisenia fetida was exposed for 28 days to the following treatments: control, double recommended field dose (2x FD) and quadruple recommended field dose (4x FD) of SHP or SKF, and mixtures of both formulations in a full factorial design. Effect criteria were mortality, reproduction, growth, respiration rate, and the activity of glutathione S-transferases (GST) and acetylcholinesterase (AChE). No significant effects on the life-cycle parameters of E. fetida, AChE activity or GST activity were observed. Therefore, chronic exposure to levels above the recommended field application doses of SHP and SKF are not expected to cause significant negative effects to the evaluated parameters in Eisenia fetida.

PollinERA aims to reverse pollinator population declines and reduce the harmful impacts of pesticides. It addresses the call through four objectives: SO1 filling ecotoxicological data gaps to enable realistic prediction of the source and routes of exposure and impact of pesticides on pollinators and their sensitivity to individual pesticides and mixtures. SO2 developing and testing a co-monitoring scheme for pesticides and pollinators across European cropping systems and landscapes, developing risk indicators and mixture exposure information. SO3 developing models for predicting pesticide toxicological effects on pollinators for chemicals and organisms, environmental fate, toxicokinetic/ toxicodynamic, and population models. SO4 developing a population-level systems-based approach to risk and policy assessment considering multiple stressors and long-term spatiotemporal dynamics at the landscape scale and generating an open database for pollinator/pesticide data and tools. This will be achieved through developing knowledge and protocols for a broad range of toxicological testing, feeding to in silico models (QSARS, toxicokinetic/toxicodynamic, and population). Using a strong stakeholder co-development approach, these models will be combined in a One System framework taking a systems view on risk assessment and policy evaluation, including an international monitoring program. The One System framework is based on EFSA’s system ERA view, expanding on the tools used for bees to include butterflies, moths and hoverflies. The consortium partners are experts in the field needed for this development and are well-placed to facilitate the uptake of tools by European bodies to guarantee the project's future impact. Expected impacts target Destination impacts of better understanding and addressing drivers of biodiversity decline, interconnected biodiversity research using digital technologies, and understanding the biodiversity and health nexus at the ecosystem level.

Although our understanding of the dramatic worldwide loss of biodiversity in recent decades is far from adequate, one of the main factors in areas dominated by agriculture is undoubtedly the widespread use of synthetic pesticides. Unfortunately, the ecological risk assessment (EcoRA) for pesticides is based on a few single-species bioassays which do not allow for the evaluation of risks to whole communities. Here we present the results of an experimental assessment of the risk to the ecosystem service provider (ESP) communities - pest control agents - from exposure to the commonly used pyrethroid insecticide, lambda-cyhalothrin. The study was performed in five European countries (Germany, Poland, Portugal, Spain, United Kingdom) representing different pedoclimatic zones. Representatives of the most common species of the ESP communities in each country were exposed in a standardized insecticide-coated glass vials bioassay to five doses of lambda-cyhalothrin: 0.8 %, 4 %, 20 %, 100 %, and 200 % of the recommended field dose (RFD) plus an untreated control. Based on the calculated LD(50)s, species sensitivity distributions (SSDs) were estimated for each country and on combined data. In all five countries, the estimated hazardous concentration for 5 % of the species (HD5) was between 0.23 % and 1.67 % RFD, with HD5 = 0.44 % RFD based on combined data. At the RFD = 7.5 g a.i./ha (active ingredient per hectare), the predicted affected fraction of the ESP communities was between 96.4 % and 99.9 % of the species (98.5 % for combined data). The results indicate an extremely high risk to ESP communities across Europe associated with the use of lambda-cyhalothrin at the recommended doses when these species are exposed to insecticide treatment. We recommend that EcoRA should include multi-species approaches, such as SSD, to better protect entire ESP communities from the negative impacts of pesticides.

Bats are an endangered group of mammals that are very sensitive to environmental stresses. One of such stress factor is trace metals pollution which threatens populations of insectivorous bats due to their top position in the food webs and exceptionally long life span. In our research Pipistrellus kuhlii was tested as a promising indicator species (urban-dwelling, sedentary, with limited daily home-range) for trace metal exposure of bats. We measured concentrations of Pb, Cu, Zn and Cd in internal and external tissues of bats from the industrial city Mariupol and the village Karlovka, Ukraine, to answer the following questions: (1) Do metal concentrations in soil samples differ between urban and rural areas, and between populations of P. kuhlii from those areas? (2) Does metal contamination differ between individuals of different sexes and ages? (3) Whether fur and/or wing membrane, the two tissues that can be collected from live bats, can be used as proxies of metal contamination in internal tissues (liver, kidney, lung, forearm bones) of P. kuhlii? Metal concentrations in soil samples were significantly higher in the city. Bats from the city accumulated significantly more Cd, Pb and Zn in external tissues than those from the rural area. Females accumulated more Cd than males, and this-year-born did not differ significantly from adult individuals. We did not find, however, significant positive correlations between metal concentrations in external and internal tissues, indicating that external tissues cannot serve as an indicator of the metal contamination of internal tissues in P. kuhlii.

Solitary bees provide an important ecological and agricultural service by pollinating both wild plants and crops, often more effectively than honey bees. In the context of worldwide pollinators' declines, it is important to better understand the functioning of populations under multiple stressors at larger spatial and temporal scales. Here we propose building a detailed, spatially-explicit agent-based model of one of the best-studied species of solitary bees, Osmia bicornis L. In this Formal Model, we review various aspects of O. bicornis biology and ecology in detail and provide descriptions of their planned implementations in the model. We also discuss the model gaps and limitations, as well as inclusions and exclusions, allowing a dialogue with the reviewers about the model's design. The ALMaSS model of O. bicornis aims to provide a realistic and detailed representation of O. bicornis populations in space and time in European agricultural landscapes. The model will be a part of the Animal, Landscape and Man Simulation System (ALMaSS); thus will be able to utilise a highly detailed, dynamic ALMaSS landscape model. It will consider the behaviour of all bee life stages daily and use state transitions to allow each individual to decide their behaviour. The development of egg-to-pupa stages in the nest will be temperature-driven. Adult bees, after they emerge from the nest in spring, will interact with the environment. They will be able to search for suitable nesting locations, provision their brood cells with pollen and reproduce. Modelled females will balance offspring size and number following the optimal allocation theory, but local environmental factors will modify their actual parental investment decisions. The model will include the daily mortality rate for the egg-to-pupa stages, overwintering mortality, and background mortality outside the nest. We will also consider the risk of open-cell parasitism as increasing with the time the brood cell is open. With the level of detail suggested, the model will be able to simulate population-level dynamics in response to multiple factors at the landscape scale over long periods. The European Food Safety Authority (EFSA) has suggested O. bicornis as a model organism for non-Apis solitary bees in the pesticide risk assessment scheme. Therefore, we hope our model will be a first step in building future landscape risk assessments for solitary bees.

The Excel files contain the results of ecotoxicological tests for the effects of selected insecticides on ESP species. The objective of this dataset is to provide original data from acute and semi-chronic laboratory tests on a few important beneficial species with broad geographic distribution. The data allow the evaluation of delayed effects and possible interactive effects of combined treatments for those pesticides that are commonly used in mixtures or sprayed next to each other in short time intervals, effectively exposing non-target arthropods to combined/sequential effects. Each data file contains the “Description” sheet where all details of the test and the exact meaning of data fields in the database are reported. The data files are named in a self-explanatory manner, starting with the name of the institution that produced the data (UC – University of Coimbra; UJA – Jagiellonian University), followed by the name of the tested species and names of tested products.

This document is composed of two parts: a report and the associated Microsoft Access database with information collected for selected Ecosystem Service Provider (ESP) species. The objective of this database was to provide compiled, and ready to use, information on the biology and ecology of key Ecosystem Service Providers (ESP) within the studied EU agricultural landscapes to be modelled in ALMaSS (agent-based or subpopulation-based models that will run on ALMaSS landscapes and will be used to test Ecostack scenarios). This information was necessary to build the “formal model” for each species, a step of paramount important in model building. The DB contains information from species being currently modelled but is prepared to be updated with new information on the same or other ESP species. To better understand the DB, it is advisable to consult the associated report.

The intensifications in the agricultural landscape and the application of pesticides can cause adverse effects on the fitness of organisms in that landscape. Here, we investigated whether habitats with different agricultural pressures influenced acetylcholinesterase (AChE) activity - a biomarker for exposure to pesticides, respiration rate, and resistance to starvation in the ground beetle Poecilus cupreus. Two differently structured landscapes were selected for the study, one dominated by small (S) and another by large (L) fields. Within each landscape three habitat types were selected: in the S landscape, these were habitats with medium (M), small (S) and no canola (meadow, 0) coverage (i.e., SM, SS, S0), and in the L landscape habitats with large (L), medium (M) and no canola (meadow, 0) coverage (i.e., LL, LM, L0), representing different levels of agricultural pressure. The activity of AChE was the highest in beetles from canola-free habitats (S0 and L0), being significantly higher than in beetles from the SM and SS habitats. The mean respiration rate corrected for body mass was also the highest in S0 and L0 beetles, with significant differences between populations from L0 vs. SS and from S0 vs. SS. Only beetles from S0, SS, L0, and LM were numerous enough to assess the resistance to starvation. Individuals from the LM habitat showed better survival compared to the canola-free habitat in the same landscape (L0), whereas in S landscape the SS beetles survived worse than those from S0, suggesting that characteristics of L landscape may lead to developing mechanisms of starvation resistance of P. cupreus in response to agricultural pressure.

The intensification of agriculture leads to increased pesticide use and significant transformation from small fields towards large-scale monocultures. This may significantly affect populations of non-target arthropods (NTA). We aimed to assess whether the multigenerational exposure to plant protection products has resulted in the evolution of resistance to insecticides in the ground beetle Poecilus cupreus originating from different agricultural landscapes. Two contrasting landscapes were selected for the study, one dominated by small and another by large fields. Within each landscape the beetles were collected at nine sites representing range of canola coverage and a variety of habitat types. Part of the collected beetles, after acclimation to laboratory conditions, were tested for sensitivity to Proteus 110 OD-the most commonly used insecticide in the studied landscapes. The rest were bred in the laboratory for two consecutive generations, and part of the beetles from each generation were also tested for sensitivity to selected insecticide. We showed that the beetles inhabiting areas with medium and large share of canola located in the landscape dominated by large fields were less sensitive to the studied insecticide. The persistence of reduced sensitivity to Proteus 110 OD for two consecutive generations indicates that either the beetles have developed resistance to the insecticide or the chronic exposure to pesticides has led to the selection of more resistant individuals naturally present in the studied populations. No increased resistance was found in the beetles from more heterogeneous landscape dominated by small fields, in which spatio-temporal diversity of crops and abundance of small, linear off-crop landscape elements may provide shelter that allows NTAs to survive without developing any, presumably costly, resistance mechanisms.

The physiological and biochemical stress induced by pesticides need to be addressed in economically and ecologically important non-Apis solitary bees, particularly at lower than field-applied concentrations. Thus, the aim of the present study was to analyse the physiological and biochemical changes in female adult Osmia bicornis bees upon continuous oral exposure to three insecticide-based agrochemicals – i.e. Dursban 480 EC (active ingredient - a.i. chlorpyrifos), Sherpa 100 EC (a.i. cypermethrin), and Mospilan 20 SP (a.i. acetamiprid), in a toxicokinetic manner (feeding with either insecticide-contaminated food or uncontaminated food (controls) for 8 d in the contamination phase followed by 8 d of decontamination (i.e. feeding with uncontaminated food)). All three tested agrochemicals altered the energetic budget of bees by the deprivation of energy derived from lipids and carbohydrates (but not proteins) and/or a decrease in respiration based metabolic rate (energy consumption) compared to the controls. The activities of acetylcholinesterase and glutathione-S-transferase enzymes were not altered by insecticides at tested concentrations. These results show that chronic exposure to at least some pesticides even at relatively low concentrations may cause severe physiological disruptions that could potentially be damaging for the solitary bees.

Agricultural landscapes have changed substantially in recent decades, shifting from the dominance of small fields (S) with diverse cropping systems toward large-scale monoculture (L), where landscape heterogeneity disappears. In this study, artificial nests of the red mason bee, Osmia bicornis, were placed in S and L landscape types on the perimeter of oilseed rape fields representing different oilseed rape coverages (ORC, % land cover). The local landscape structure around each nest was characterised within a 100, 200, 500, and 1000 m radius using ORC and 14 landscape characteristics, which were then reduced by non-metric multidimensional scaling (nMDS) to two axes: nMDS1 characterised the dataset primarily according to land fragmentation and the main crop, whereas nMDS2 captured the prevalence of more natural areas in the landscape. Pollen diversity and insecticide risk levels in the pollen provisions collected by the bees were analysed, and their dependence on the landscape structure was tested. Thereafter, the effects of pollen diversity, insecticide risk, and landscape structure on the life-history traits of bees and their sensitivity to topically applied Dursban 480 EC were determined. Pollen taxa richness in a single nest ranged from 3 to 12, and 34 pesticides were detected in the pollen at concentrations of up to 320 ng/g for desmedipham. The O. bicornis foraging range was relatively large, indicating that the landscape structure within a radius of ~1000 m around the nest is important for this species. Pollen diversity in the studied areas was of minor importance for bee performance, but the ORC or landscape structure significantly affected the life-history traits of the bees. Contamination of pollen with insecticides affected the bees by decreasing the mass of newly emerged adults but their sensitivity to Dursban 480 EC was not related to environmental variables.

The landscape structure of an agro-ecosystem can directly influence its biodiversity. In order to better understand the ongoing worldwide biodiversity collapse, a comprehensive landscape characterisation method is indispensable. In this paper we propose a robust and universal methodological approach to describe agricultural landscape properties tailored for the species of interest. In the proposed method we advocate for using species-specific classes of landscape elements and proper spatial scale reflecting the species home range. As a result, complex landscape description is reduced to relatively few main landscape shaping factors that can serve as landscape metavariables in any further analysis of choice. As a working example, we used a non-metric multidimensional scaling-based method to evaluate how the landscape-level spatial properties of two distinct agricultural landscapes – dominated either by large-scale industrial agriculture or by traditional small-scale farming – shape the environment available to the red mason bee (Osmia bicornis) at different spatial scales. The results for the landscapes studied herein suggest that for pollinators with a home range radius above ca. 100 m, not only local habitat counts but also large-scale landscape properties are of prime importance. This means that for a successful management of biodiversity of such species, large-scale landscape structure must be considered.

All European bats are protected by the EU and Associated Members legal regulations. Being insectivorous and top predators, they can be particularly exposed to persistent organic and inorganic pollutants. It is surprising how little is known about the impact of environmental pollutants on bats from physiological to populational levels. In this study we focused on contamination with trace metals of first-year bats from Kharkiv city, NE Ukraine. Tissues from the carcasses of two species, Nyctalus noctula (n = 20) and Eptesicus serotinus (n = 20), were used for metal analysis. The samples of external (wing membrane, fur) and internal (liver, lung, kidney, bones) tissues were analysed for contents of Pb, Cu, Zn, and Cd to see whether fur or wing membrane can be used as proxies for metal contamination of the vital internal tissues. In E. serotinus, significant positive correlations in Pb concentrations were found between all external and internal tissues. For Cd only, correlation between the fur and lung was found, for Cu between the fur and liver, and for Zn between the fur and kidney. In contrast, for N. noctula, only one such correlation was found - between Zn concentrations in the fur and kidney. The tissues differed significantly in concentrations of all studied metals, with no difference between the species. The results showed that the fur and wing membrane can be used as good proxies for Pb concentrations in internal organs of E. serotinus, but not necessarily for other metals or for N. noctula. The results for Pb are, however, encouraging enough to conclude that the topic is worth further studies, covering more species, a wider age range and more diverse environments. (C) 2021 The Authors. Published by Elsevier Ltd.