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A growing body of scientific literature stresses the need to advance current environmental risk assessment (ERA) methodologies and associated regulatory frameworks to better address the landscape-scale and long-term impact of pesticide use on biodiversity and the ecosystem. Moreover, more collaborative and integrative approaches are needed to meet sustainability goals. The One Health approach is increasingly applied by the European Food Safety Authority (EFSA) to support the transition towards safer, healthier and more sustainable food. To this end, EFSA commissioned the development of a roadmap for action to establish a European Partnership for next-generation, systems-based Environmental Risk Assessment (PERA). Here, we summarise the main conclusions and recommendations reported in the 2022 PERA Roadmap. This roadmap highlights that fragmentation of data, knowledge and expertise across regulatory sectors results in suboptimal processes and hinders the implementation of integrative ERA approaches needed to better protect the environment. To advance ERA, we revisited the underlying assumptions of the current ERA paradigm; that chemical risks are generally assessed and managed in isolation with a substance-by-substance, realistic worst-case and tiered approach. We suggest optimising the use of the vast amount of information and expertise available with pesticides as a pilot area. It is recommended to as soon as possible adopt a systems-based approach, i.e. within the current regulatory framework, to spark a step-wise transition towards an ERA framed at a system level of ecological and societal relevance. Tangible systems-based and integrative steps are available. For instance, the rich sources of existing data for prospective and retrospective ERA of pesticides could be used to reality-benchmark existing and new ERA methods. To achieve these goals, collaboration among stakeholders across scientific disciplines and regulatory sectors must be strengthened., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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, λ-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 λ-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 λ-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., Competing Interests: Declaration of competing interest Ryszard Laskowski reports financial support was provided by European Union. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

In this study we aimed to assess the role of wild red deer, along with other ungulates such as roe deer and wild boar, in the soil's physical properties, namely soil penetration resistance and depth (used as a proxy for soil compaction), hydraulic conductivity (a proxy for water infiltration), and the proportion of soil stable aggregates. Results showed that, at the density level found in our study area, red deer have a neutral effect at the soil level, not causing significant soil compaction or significantly influencing measured soil functions., (© 2024 International Society of Zoological Sciences, Institute of Zoology/Chinese Academy of Sciences and John Wiley & Sons Australia, Ltd.)

While pesticide use is subject to strict regulatory oversight worldwide, it remains a main concern for environmental protection, including biodiversity conservation. This is partly due to the current regulatory approach that relies on separate assessments for each single pesticide, crop use, and non-target organism group at local scales. Such assessments tend to overlook the combined effects of overall pesticide usage at larger spatial scales. Integrative landscape-based approaches are emerging, enabling the consideration of agricultural management, the environmental characteristics, and the combined effects of pesticides applied in a same or in different crops within an area. These developments offer the opportunity to deliver informative risk predictions relevant for different decision contexts including their connection to larger spatial scales and to combine environmental risks of pesticides, with those from other environmental stressors. We discuss the needs, challenges, opportunities and available tools for implementing landscape-based approaches for prospective and retrospective pesticide Environmental Risk Assessments (ERA). A set of "building blocks" that emerged from the discussions have been integrated into a conceptual framework. The framework includes elements to facilitate its implementation, in particular: flexibility to address the needs of relevant users and stakeholders; means to address the inherent complexity of environmental systems; connections to make use of and integrate data derived from monitoring programs; and options for validation and approaches to facilitate future use in a regulatory context. The conceptual model can be applied to existing ERA methodologies, facilitating its comparability, and highlighting interoperability drivers at landscape level. The benefits of landscape-based pesticide ERA extend beyond regulation. Linking and validating risk predictions with relevant environmental impacts under a solid science-based approach will support the setting of protection goals and the formulation of sustainable agricultural strategies. Moreover, landscape ERA offers a communication tool on realistic pesticide impacts in a multistressors environment for stakeholders and citizens., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Implementation of marker-assisted selection (MAS) in modern beekeeping would improve sustainability, especially in breeding programs aiming for resilience against the parasitic mite Varroa destructor . Selecting honey bee colonies for natural resistance traits, such as brood-intrinsic suppression of varroa mite reproduction, reduces the use of chemical acaricides while respecting local adaptation. In 2019, eight genomic variants associated with varroa non-reproduction in drone brood were discovered in a single colony from the Amsterdam Water Dune population in the Netherlands. Recently, a new study tested the applicability of these eight genetic variants for the same phenotype on a population-wide scale in Flanders, Belgium. As the properties of some variants varied between the two studies, one hypothesized that the difference in genetic ancestry of the sampled colonies may underly these contribution shifts. In order to frame this, we determined the allele frequencies of the eight genetic variants in more than 360 Apis mellifera colonies across the European continent and found that variant type allele frequencies of these variants are primarily related to the A. mellifera subspecies or phylogenetic honey bee lineage. Our results confirm that population-specific genetic markers should always be evaluated in a new population prior to using them in MAS programs.

Laboratory ecotoxicological tests are important tools for the management of environmental changes derived from anthropogenic activities. Folsomia candida is usually the model species used in some procedures. However, this species may not be sufficiently representative of the sensitivity of the other collembolan species. This study aimed to evaluate (i) the effects of soils naturally rich in potentially toxic elements (PTE) and soil characteristics on the reproduction and survival of different collembolan species, (ii) whether the habitat function of these soils is compromised, and (iii) to what extent F. candida is representative of the other collembolan species. For this, reproduction tests with six collembolan species were conducted in 14 different samples of soils. In general, collembolan reproduction was not completely inhibited in none of the natural tested soils. Even soils with high pollution load index values did not negatively affect collembolan reproduction for most of the species. In contrast, the lowest collembolan reproduction rates were found in a visually dense soil (lowest volume/weight ratio), highlighting that soil attributes other than total PTE concentration also interfere in the reproduction of collembolans. Our results support the idea that the F. candida species might not be representative of other collembolan species and that laboratory tests to assess soil contaminations should be conducted using diverse collembolan species., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

In this work we aim to provide a quantitative method allowing the probing of the physiological status of honeybee colonies by providing them with a gentle, short, external artificial vibrational shockwave, and recording their response. The knock is provided by an external electromagnetic shaker attached to the outer wall of a hive, driven by a computer with a 0.1 s long, monochromatic vibration at 340Hz set to an amplitude that occasionally yields a mild response from the bees, recorded by an accelerometer placed in the middle of the central frame of the colony. To avoid habituation, the stimulus is supplied at randomised times, approximately every hour. The method is pioneered with a pilot study on a single colony hosted indoors, then extended onto eight outdoors colonies. The results show that we can quantitatively sense the colony's overall mobility, independently from another physiological aspect, which is phenomenologically explored. Using this, a colony that is queenless is easily discriminated from the others., (© 2024. The Author(s).)

Honey bee colonies have great societal and economic importance. The main challenge that beekeepers face is keeping bee colonies healthy under ever-changing environmental conditions. In the past two decades, beekeepers that manage colonies of Western honey bees ( Apis mellifera ) have become increasingly concerned by the presence of parasites and pathogens affecting the bees, the reduction in pollen and nectar availability, and the colonies' exposure to pesticides, among others. Hence, beekeepers need to know the health condition of their colonies and how to keep them alive and thriving, which creates a need for a new holistic data collection method to harmonize the flow of information from various sources that can be linked at the colony level for different health determinants, such as bee colony, environmental, socioeconomic, and genetic statuses. For this purpose, we have developed and implemented the B-GOOD (Giving Beekeeping Guidance by computational-assisted Decision Making) project as a case study to categorize the colony's health condition and find a Health Status Index (HSI). Using a 3-tier setup guided by work plans and standardized protocols, we have collected data from inside the colonies (amount of brood, disease load, honey harvest, etc.) and from their environment (floral resource availability). Most of the project's data was automatically collected by the BEEP Base Sensor System. This continuous stream of data served as the basis to determine and validate an algorithm to calculate the HSI using machine learning. In this article, we share our insights on this holistic methodology and also highlight the importance of using a standardized data language to increase the compatibility between different current and future studies. We argue that the combined management of big data will be an essential building block in the development of targeted guidance for beekeepers and for the future of sustainable beekeeping.

Basamid® is a fumigant nematicide and fungicide known to break down in several volatile compounds, mainly methyl isothiocyanate (MITC), when in contact with water. Soil abiotic parameters, such as pH, influences this breakdown process, and thus, the toxic effects of Basamid® to aquatic biota. This work studied the influence of soil pH (5.5, 6.5 and 7.5) on the toxicity of eluates (1:4, m:v), obtained from Basamid®-contaminated soils (with the recommended dose of 145 mg of dazomet/Kg of soil), on two primary consumers: Daphnia magna and Brachionus calyciflorus. For this, lethal and sublethal toxicity of eluates originated from soils at pH 5.5, 6.5 and 7.5, contaminated with Basamid® (Ba-E 5.5; 6.5 and 7.5, respectively), were assessed (dilutions between 0.096 - 100%). The LD 50,24h of Basamid® eluates for D. magna varied from 3.07% to 7.82% (Ba-E 6.5 and Ba-E 5.5 respectively), while for B. calyciflorus varied from 18.1% to 84.7% (Ba-E 6.5 and Ba-E 7.5, respectively). Both species were less sensitive to Basamid® eluates originated from soils with pH 7.5 and more sensitive to those obtained from soils with pH 6.5. Regarding the sublethal effects, a lower soil pH was associated with a higher toxicity of Basamid® to D. magna reproduction (LOED: 0.125% Ba-E 5.5), while for B. calyciflorus such a higher toxicity was observed at the highest soil pH (ED 20 : 7.42% [5.10-9.74] at Ba-E 7.5). These results show a negative association between soil pH and the lethal toxicity of Basamid® contaminated eluates. However, such a pattern was not observed at sublethal level, at which a species dependency was observed regarding the influence of soil pH in the observed toxicity. Nevertheless, it is to highlight that very low concentrations of eluates (as 3.07%) caused significant mortality, indicating a high risk for freshwater biota. Considering that Basamid® is likely to reach the aquatic systems is real, for which reason the recommended dose must be reviewed at environmentally-relevant scenarios., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

The COVID-19 pandemic has drastically affected people's social habits, especially those related to outdoor activities. We intended to understand the effects of the two national lockdowns in Portugal on the presence and activity of a wild population of red deer ( Cervus elaphus ) by analysing data from camera traps installed at Lousã mountain, in the central part of Portugal. The cameras were set between 2019 and 2021, and a total of 2434 individual contacts of red deer and 182 contacts of people were recorded. Results showed a higher human presence in the mountain area during the COVID-19 outbreak, especially during the first lockdown in 2020 (0.05 ± 0.17 individuals/day), compared to the same period of the year before the pandemic (0.02 ± 0.05 individuals/day), which resulted in an increase of people by 150%. The increase in human presence did not have a significant direct effect on the presence of red deer. Despite the low overlap of activity patterns between people and red deer, deer showed avoidance behaviour in the 24 h after the detection of human presence on camera traps, as well as an increase in daily activity during the 2020 lockdown, showing red deer's awareness of human visitation. These results showed that people's increased search for cultural services in wild environments during COVID-19 lockdowns, such as hiking and biking, seemed to influence the population of red deer, albeit momentarily., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 Published by Elsevier B.V.)

Agrochemicals are mostly used to deplete pests and treat diseases in terrestrial agro-ecosystems. However, their transport through the soil, by leaching and/or runoff, may cause them to reach aquatic systems. Environmental parameters, such as soil pH, can affect this transport, by influencing the magnitude of agrochemicals degradation and chemical reaction. This work aimed at investigating the influence of soil pH on the toxicity of eluates obtained from Basamid® contaminated soils to Hydra viridissima, Xenopus laevis and Danio rerio. For this, a natural soil with pH amended to 5.5, 6.5 and 7.5, was spiked with the recommended dose (RD) of Basamid® (145 mg dazomet/kg soil) and eluates (Ba-E) were prepared with the respective species culture medium. Dilutions of the eluates (0.14-100%), obtained from the three soils (Ba-E 5.5, Ba-E 6.5 and Ba-E 7.5, corresponding to soil spiked with Basamid® RD at soil pH of 5.5, 6.5 and 7.5, respectively), were used to expose the organisms. Results showed that for H. viridissima increased soil alkalinity provoked less mortality comparatively to lower soil pH [LD 50,96h of Ba-E 5.5: 10.6% and LD 50,96h of Ba-E 7.5: 21.2%]. As for X. laevis and D. rerio Ba-E lethal ecotoxicity was similar across soil pH (LD 50,96h varied from 5.7 to 6.9% and from 2.1 to 4.3%, respectively). For malformations, 20% effect dilution (ED) in H. viridissima was significantly higher at Ba-E 7.5 (ED 20,96h : 17.4%), comparatively to Ba-E 5.5 and Ba-E 6.5 (ED 20,96h : 7.9% and 7.7%, respectively). From the three tested organisms and based on both lethal and sublethal effects, H. viridissima presented the highest tolerance to Basamid® eluates and soil pH was a major factor determining the fumigant toxicity, with higher soil pH levels inducing, lower toxicity. The eluates obtained from soils contaminated with RD of Basamid® induced severe effects to the three aquatic species., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Sexual segregation is a common phenomenon among animals, particularly dimorphic ones. Although widely addressed, the reasons and consequences of sexual segregation are still an important topic in need of better understanding. In this study, we mainly evaluate the diet composition and feeding behaviour of animals, which are related to the use of different habitats by the sexes, a special case of sexual segregation also termed habitat segregation. Sexually size dimorphic males and females often have different energetic and nutritional needs and, thus, different diets. We collected fresh faecal samples from wild Iberian red deer ( Cervus elaphus L.) in Portugal. Samples were analysed in terms of diet composition and quality. As expected, both sexes differed in their diet composition, with males eating more arboreous species than females, but this difference was affected by sampling periods. Diet composition of both sexes had the biggest differences (and the lowest overlap) in spring, which corresponds to the end of gestation and beginning of birth. These differences might be a consequence of the sexual body size dimorphism characteristic of this species, as well as of different needs due to different reproductive costs. No differences regarding the quality of the excreted diet were observed. Our results may help to understand some patterns of sexual segregation observed in this red deer population. However, besides foraging ecology, other factors may also be contributing to sexual segregation in this Mediterranean population of red deer, and further studies focusing on sexual differences regarding feeding behaviour and digestibility are needed.

Enhanced drought, more frequent rainfall events and increased inter-annual variability of precipitation are the main trends of climate expected for the Mediterranean. Drought is one of the most important stressors for plants and significantly impacts plant communities causing changes in plant composition and species dominance. Through an experiment under controlled conditions, we assessed the response of Mediterranean species from different functional groups (annual grass, annual forb, annual legume, and perennial shrub) to moderate and severe water deficit. Changes in plant traits (leaf dry matter), biomass and physiology (water status, photosynthesis, pigments, and carbohydrate) were evaluated. The studied species differed in their response to water deficit. Ornithopus compressus , the legume, showed the strongest response, particularly under severe conditions, decreasing leaf relative water content (RWC), pigments and carbohydrates. The grass, Agrostis pourreti and the forb, Tolpis barbata , maintained RWC, indicating a higher ability to cope with water deficit. Finally, the shrub, Cistus salviifolius , had the lowest response to stress, showing a higher ability to cope with water deficit. Despite different responses, plant biomass was negatively affected by severe water deficit in all species. These data provide background for predicting plant diversity and species composition of Mediterranean grasslands and Montado under climate change conditions.

Intensive agriculture along with the use of agrochemicals has been associated with low soil fertility, soil erosion, and soil acidity. Management of soil pH through liming is a common practice in agriculture to increase soil fertility and nutrient availability. When altering soil pH, different chemical reactions occur depending on soil composition and agrochemicals presence. Basamid® is a fumigant used worldwide targeting soil nematodes, fungi, and weeds in diverse crops, that can reach freshwater ecosystems by leaching through the soil layers. The major goal of this work was to assess the influence of soil pH in the toxicity of Basamid® eluates to the microalgae Raphidocelis subcapitata and the duckweed Lemna minor. For this, eluates were prepared from soils with different pH (5.5, 6.5 and 7.5), contaminated with the recommended dose of Basamid® corresponding to 145.7 mg of dazomet/Kg soil. Soil was amended with calcium carbonate (CaCO 3 ). Raphidocelis subcapitata and L. minor were exposed to the eluates during 72 h and 7 days respectively, and multiple endpoints were assessed: growth rate, biomass, pigment as chlorophyl content and cell damage. Results showed that soil pH can influence the performance of the tested species and also be a major factor in influencing Basamid®'s toxicity. However, a clear pattern of the influence of soil pH on Basamid®'s toxicity was not observed and was species dependent. For R. subcapitata lower soil pHs induced higher toxicity of Basamid®'s to the algae [ED 50 for growth rate: 30 % (confidence limits-CL: 22.8-37.2) for soil pH 5.5; >100 % for soil pH 6.5 and pH 7.5], while for L. minor the opposite was observed [ED 50 for number of fronds: 27.2 % (CL: 22.8-31.6) for pH 5.5; 20.3 % (CL: 10.0-30.6) for pH 6.5 and 10.7 % (CL: 6.3-15.1)]. Overall, these results showed that leachates of Basamid® through soils, at recommended doses, can have a severe impact on aquatic systems, with or without the influence of abiotic factors., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Eucalyptus plantations occupy 26 % of Portuguese forested areas. Its flowers constitute important sources for bees and beekeepers take advantage of this and keep their honey bee colonies within or near the plantations for honey production. Nonetheless, these plantations are susceptible to pests, such as the eucalyptus weevil Gonipterus platensis. To control this weevil, some plantations must be treated with pesticides, which might harm non-target organisms. This study aimed to perform a multifactorial assessment of the health status and development of Apis mellifera iberiensis colonies in two similar landscape windows dominated by Eucalyptus globulus plantations - one used as control and the other with insecticide treatment. In each of the two selected areas, an apiary with five hives was installed and monitored before and after a single application of the insecticide acetamiprid (40 g a.i./ha). Colony health and development, resources use, and pesticide residues accumulation were measured. The results showed that the application of acetamiprid in this area did not alter the health status and development of the colonies. This can be explained by the low levels of residues of acetamiprid detected only in pollen and bee bread samples, ~52 fold lower than the sublethal effect threshold. This could be attributed to the low offer of resources during and after the application event and within the application area, with the consequent foraging outside the sprayed area during that period. Since exposure to pesticides in such complex landscapes seems to be dependent on the spatial and temporal distribution of resources, we highlight some key monitoring parameters and tools that are able to provide reliable information on colony development and use of resources. These tools can be easily applied and can provide a better decision-taking of pesticide application in intensive production systems to decrease the risk of exposure for honey bees., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Honey bee colonies have shown abnormal mortality rates over the last decades. Colonies are exposed to biotic and abiotic stressors including landscape changes caused by human pressure. Modern agriculture and even forestry, rely on pesticide inputs and these chemicals have been indicated as one of the major causes for colony losses. Neonicotinoids are a common class of pesticides used worldwide that are specific to kill insect pests, with acetamiprid being the only neonicotinoid allowed to be applied outdoors in the EU. To evaluate honeybees' exposure to acetamiprid under field conditions as well as to test the use of in-situ tools to monitor pesticide residues, two honeybee colonies were installed in five Eucalyptus sp. plantations having different area where Epik® (active substance: acetamiprid) was applied as in a common spraying event to control the eucalyptus weevil pest. Flowers, fresh nectar, honey bees and colony products samples were collected and analyzed for the presence of acetamiprid residues. Our main findings were that (1) acetamiprid residues were found in samples collected outside the spraying area, (2) the amount of residues transported into the colonies increased with the size of the sprayed area, (3) according to the calculated Exposure to Toxicity Ratio (ETR) values, spraying up to 22 % of honeybees foraging area does not harm the colonies, (4) colony products can be used as a valid tool to monitor colony accumulation of acetamiprid and (5) the use of Lateral Flow Devices (LFDs) can be a cheap, fast and easy tool to apply in the field, to evaluate the presence of acetamiprid residues in the landscape and colony products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Understanding the effects of forest-to-agriculture conversion on microbial diversity has been a major goal in soil ecological studies. However, linking community assembly to the ruling ecological processes at local and regional scales remains challenging. Here, we evaluated bacterial community assembly patterns and the ecological processes governing niche specialization in a gradient of geography, seasonality, and land-use change, totaling 324 soil samples, 43 habitat characteristics (abiotic factors), and 16 metabolic and co-occurrence patterns (biotic factors), in the Brazilian Atlantic Rainforest, a subtropical biome recognized as one the world's largest and most threatened hotspots of biodiversity. Pairwise beta diversities were lower in pastures than in forest and no-till soils. Pasture communities showed a predominantly neutral model, regarding stochastic processes, with moderate dispersion, leading to biotic homogenization. Most no-till and forest microbial communities followed a niche-based model, with low rates of dispersal and weak homogenizing selection, indicating niche specialization or variable selection. Historical and evolutionary contingencies, as represented by soil type, season, and dispersal limitation were the main drivers of microbial assembly and processes at the local scale, markedly correlated with the occurrence of endemic microbes. Our results indicate that the patterns of assembly and their governing processes are dependent on the niche occupancy of the taxa evaluated (generalists or specialists). They are also more correlated with historical and evolutionary contingencies and the interactions among taxa (i.e., co-occurrence patterns) than the land-use change itself., (© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Agricultural intensification has increased the number of stressors that pollinators are exposed to. Besides increasing landscape fragmentation that limit the supply of flower resources, intensive agricultural practices relying on the use of pesticides to control agricultural pests also affect non-target organisms like honey bees. The use of most pesticides containing neonicotinoids has been severely restricted in the European Union, leaving pesticides containing acetamiprid as the only ones that are still authorized. In the meantime, new substances like sulfoxaflor, that have a similar mode of action acting on the insect's nicotinic acetylcholine receptors (nAChR), have been approved for agricultural use. In Europe and USA, the use of pesticides containing this active ingredient is limited due to toxic effects already reported on bees, but no restrictions regarding this matter were applied in other countries (e.g., Brazil). In this study, homing ability tests with acetamiprid and sulfoxaflor were performed, in which honey bees were fed with three sub-lethal doses from each substance. After exposure, each honey bee was equipped with an RFID chip and released 1 km away from the colony to evaluate their homing ability. No significant effects were detected in honey bees fed with 32, 48 and 61 ng of acetamiprid while a poor performance on their homing ability, with only 28% of them reaching the colony instead of 75%, was detected at a 26 ng/a.s./bee dose of sulfoxaflor. Although, both pesticides act on the nAChR, the higher sulfoxaflor toxicity might be related with the honey bees detoxifying mechanisms, which are more effective on cyano-based neonicotinoids (i.e., acetamiprid) than sulfoximines. With this study we encourage the use of homing ability tests to be a suitable candidate to integrate the future risk assessment scheme, providing valuable data to models predicting effects on colony health that emerge from the individual actions of each bee., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Climate change may interfere with the behavior of pesticides and organisms, influencing pesticides toxicity to non-target organisms like collembolans. Aiming to assess the representativeness of the standardized species F. candida to the Collembola group under different temperatures, four species of collembolans - Folsomia candida, Folsomia fimetaria, Proisotoma minuta and Sinella curviseta - were exposed to a new generation insecticide of Chlorantraniliprole, under the standardized temperature of 20 °C, and a temperature foreseeing a global warming scenario of 25 °C. Results showed that F. candida, F. fimetaria and P. minuta were sensitive to Chlorantraniliprole at both temperatures, while S. curviseta was insensitive to the insecticide concentrations up to 457 mg a.i./kg of soil, regardless of the temperature. The sensitivity of F. candida and P. minuta was significantly higher at 25 °C than at 20 °C, while F. fimetaria and S. curviseta remained equally sensitive/insensitive to both temperatures. Results suggest that F. candida can be representative of F. fimetaria under standard conditions but not for F. fimetaria under 25 °C nor for P. minuta and S. curviseta under both temperatures due to the higher sensitivity of F. candida. On the other hand, due to its higher sensitivity, F. candida can be used to define environmentally protective measures (at both test temperatures) but the use of additional Collembola species is recommended to avoid the definition of over-protective goals., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Natal-da-Luz, Tiago reports financial support was provided by H2020 Food Security Sustainable Agriculture and Forestry Marine Maritime and Inland Water Research and the Bioeconomy. Has patent pending to., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Stream intermittency - periodic sequences of water flow cessation and resumption - occurs throughout the year, across seasons. Even though temperature is a known regulator of litter decomposition in both terrestrial and aquatic environments, comparative experiments on drought durations at distinct temperatures on microbial-mediated decomposition in streams experiencing intermittency are still lacking. Here, three drought temperatures (5, 15 and 25 °C) and two durations (short: 2.5 weeks; long: 5 weeks) were applied in a microcosm study to oak leaf discs colonized in a reference stream; mass loss and associated microbial parameters (fungal biomass, microbial activity, and sporulation rates) were evaluated following re-submersion for 2 weeks. Higher mass loss was found at 15 °C than 25 °C. A prolongation of the drought exposure period had no effect on mass loss, suggesting an early (≤ 2.5 weeks) inhibitor effect of drought on microbial-mediated leaf degradation. Fungal biomass was highest at 25 °C following a short drought, and decreased with a longer drought period at both 15 °C and 25 °C. Microbial activity was not affected by either drought duration or temperature. Sporulation rates and fungal diversity were significantly reduced by the longer drought period; in the short treatment, maximum values were found at 15 °C. In contrast to longer droughts, aquatic fungal communities during short dry periods seem to invest in energetically-expensive physiological responses to desiccation (e.g., ergosterol production) promoting biomass accrual at the expense of mass loss and reproductive output. Under more severe desiccation (higher duration and temperature), the lower diversity of fungal communities seem to result in negative legacy effects for fungal growth and reproductive capacity after flow resumption. These results suggest that native riparian vegetation, through its ability to regulate temperature in streams, may be critical in protecting freshwaters from intensified severity of drought periods in streams experiencing intermittency., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Biobased and biodegradable plastic mulch films (aka, mulch biofilm) have emerged as a sustainable alternative to conventional plastic mulch films in agriculture, promising to reduce soil contamination with plastic residues through in situ biodegradation. However, current standards certifying biodegradable plastics cannot predict biodegradability in natural settings. The scarce studies considering the possible biodegradation and ecotoxicity of mulch biofilms in soil systems question the environmental friendliness of these alternative options. This study assessed the biodegradation of a commercially available mulch biofilm by the soil-dwelling fungus Penicillium brevicompactum (in solid culture media and soil for 15 and 28 days, respectively), and the ecotoxicological effects of mulch biofilm microplastics on the earthworm Eisenia andrei (pristine or UV-weathered, at 0.125-0.250-0.500 g/kg). Results (from microplastics' mass loss, microscopy, and FTIR spectroscopy) suggest that the presence of P. brevicompactum promotes mulch biofilm's biodegradation. Exposure to environmental concentrations of pristine biofilm microplastics (and its ingestion) increased earthworms' sensitivity to touch, induced physiological alterations, decreased energy reserves, and decreased their reproduction (>30%). Conversely, exposure to weathered biofilm microplastics slightly increased earthworms' sensitivity, as well as carbohydrate reserves,without affecting their reproduction. The tested mulch biofilm seems to be, at first sight, an environmentally friendly alternative as it presented susceptibility for biodegradation by a widespread fungus, and the absence of ecotoxicological chronic effects on a key macroinvertebrate species in soil ecosystems when considering environmental relevant concentrations and plastics weathered conditions. Notwithstanding, the obtained results highlight the need to revise current standards, as they often neglect the role of, and their chronic effects on, naturally occurring organisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

In this study the recolonization concentration concept for soil organisms is presented and validated. This concept is based on the empirically deduced avoidance-recolonization hypothesis, which shows a negative correlation between avoidance (AC x ) and recolonization (RC x ) (AC x = RC 100-x ) responses. The concept was validated in a two-step approach composed by (i) individual placement tests, to demonstrate the non-influence of individual placement in a dual chamber avoidance test and (ii) small scale gradient tests to demonstrate that the number of colonizers reaching a soil patch with a certain concentration is independent on their previous exposure to lower concentrations. Overall, data show that avoidance data can be used, when framed under the recolonization concentration concept, to evaluate the recolonization potential of contaminated sites. The recolonization concept is an important theoretical concept that when coupled with spatial modelling tools could be used to tackle the spatial and temporal recovery dynamics of contaminated soil.

Soils are habitat to a variety of flora and fauna in a linked ecosystem which provides essential ecosystem services. In soil, metals can accumulate at high concentrations, because of anthropogenic activities, leading to toxic effects, threatening the ecosystem and the services it provides. In most real-world contamination scenarios, metals occur as complex mixtures which can interact and produce different toxicity than predicted from individual metal data. Current regulatory guidelines are based on single species responses to individual metals and ignore indirect effects inherent to the inter-linked nature of ecosystems. Also, the evaluation of anthropogenic impacts to the soil communities is usually measured through structural endpoints (e.g. abundance) disregarding functional measurements (e.g. organic matter decomposition rates), which are often seen as tightly related, and thus, similarly affected. In this study we tested three mixture ratios of five metal oxides (lead, copper, nickel, zinc, cobalt) at three dose levels (Low, Med, High) in a terrestrial model ecosystem experiment and measured structural and functional endpoints. Exposure to metal mixtures for 16 weeks did not affect the microarthropod community, but produced severe effects on soil microbial activity (PNR and DHA) reducing activity below 50% compared to control levels, in all dosed treatments. Metal contamination also significantly affected feeding activity and organic matter decomposition, but effects were not as pronounced as on microbial activity. Data suggest that, in the risk assessment of metals and their mixtures, effects on ecosystem structure and functions must be considered to provide adequate environmental protection., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Information concerning arbuscular mycorrhizal (AM) fungal geographical distribution in tropical and subtropical soils from the Atlantic Forest (a global hotspot of biodiversity) are scarce and often restricted to the evaluation of richness and abundance of AM fungal species at specific ecosystems or local landscapes. In this study, we hypothesized that AM fungal diversity and community composition in subtropical soils would display fundamental differences in their geographical patterns, shaped by spatial distance and land-use change, at local and regional scales. AM fungal community composition was examined by spore-based taxonomic analysis, using soil trap cultures. Acaulospora koskei and Glomus were found as generalists, regardless of mesoregions and land uses. Other Acaulospora species were also found generalists within mesoregions. Land-use change and intensification did not influence AM fungal composition, partially rejecting our first hypothesis. We then calculated the distance-decay of similarities among pairs of AM fungal communities and the distance-decay relationship within and over mesoregions. We also performed the Mantel test and redundancy analysis to discriminate the main environmental drivers of AM fungal diversity and composition turnover. Overall, we found significant distance-decays for all land uses. We also observed a distance-decay relationship within the mesoregion scale (< 104 km) and these changes were correlated mainly to soil type (not land use), with the secondary influence of both total organic carbon and clay contents. AM fungal species distribution presented significant distance-decays, regardless of land uses, which was indicative of dispersal limitation, a stochastic neutral process. Although, we found evidence that, coupled with dispersal limitation, niche differentiation also played a role in structuring AM fungal communities, driven by long-term historical contingencies, as represented by soil type, resulting from different soil origin and mineralogy across mesoregions., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC part of Springer Nature.)

The goal of this study was to incorporate community data into the effect assessment of environmental and regulatory relevant metal mixtures. In this experiment three fixed mixture ratios (Canadian soil quality guideline ratio - CSQG; Agricultural, residential and Loamy ratio - ARL; and Sudbury ratio - SUD) were tested in a natural community microcosm with 11 doses for each mixture ratio. The effect of metal mixtures on the community was measured using the community effect concentration (EC) concept which assumes that as contamination increases, the community similarity between test and control treatments decreases producing a dose response curve allowing the calculation of community effect concentrations. In regulatory mixture ratios (CSQG and ARL) community EC10s were four times higher than regulatory thresholds and current regulation might be overprotective of the microarthropod communities in some soils. For the contaminated site ratio (SUD), the field dose in the contaminated site corresponded to a community EC20 and if metal concentrations were reduced by 1TU, (from 3.1TU to 2.1TU) effects would be below a community EC10. Overall, the community EC concept was successfully applied and has the potential for inclusion in risk assessment schemes as a measure of community response., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The contamination left by abandoned mines demands sustainable mitigation measures. Hence, the aim of this study was to examine the phytoremediator ability of Eucalyptus globulus Labill. to be used for cleaning up metal-contaminated soils from an African abandoned iron (Fe) mine (Ait Ammar, Oued Zem, Morocco). Plantlets of this species were exposed to a control (CTL), a reference (REF), and a mine-contaminated soil (CS). Morphological (growth, leaf area) and physiological stress biomarkers (photosynthetic efficiency, pigments content, leaf relative water, and malondialdehyde (MDA) levels) and metal bioaccumulation were assessed. The growth and leaf area of E. globulus increased overtime in all soils, although at a lower rate in the CS. Its photosynthetic efficiency was not markedly impaired, as well as MDA levels decreased throughout the experiment in CS. In this soil, higher metal contents were detected in E. globulus roots than in leaves, especially Fe (roots: 15.98-213.99 μg g -1 ; leaves: 5.97-15.98 μg g -1 ) and Zn (roots: 1.64-1.99 μg g -1 ; leaves: 0.67-1.19 μg g -1 ), indicating their reduced translocation. Additionally, though at low extent, the plants bioaccumulated some metals (Pb > Zn > Cu) from CS. Overall, E. globulus may be potentially used for the phytoremediation of metals in metal-contaminated soils.

Olive growing has been intensified through the simplification of agricultural landscapes. In order to rethink the environmental drawbacks of these practices, conservation biological control techniques have been examined. In this work, Prays oleae and its natural enemy Chrysoperla carnea were monitored to account for the effects of the amount and diversity of different land-uses. We found that C. carnea showed an attraction to areas with high abundances of P. oleae but this predator did not display any affection by the different land-uses. Inversely, P. oleae abundance was lower in diverse landscapes and higher in simplified ones. Importantly, higher abundances of C. carnea were related to lower infestation levels of P. oleae in the late part of the season. These results corroborate the attraction of C. carnea to the olive moth, highlighting the potential of C. carnea as a biological control agent of this pest, assert that the promotion of land-use diversity can reduce P. oleae and confirm that landscapes dominated by olive groves can promote this pest. The present study aims at contributing to the discussion about the management of agricultural ecosystems by providing farmers with sustainable alternatives that do not have harmful effects on the environment and public health.

Phosphorus is a critical element to agriculture, consequently global phosphate rock demand will remain rising to feed a growing world population. The beneficiation of phosphorous ore gives rise to several tons of a waste by-product [phosphogypsum (PG)] which valorisation is limited, within other reasons, by the risks posed to environment and human health. Although threatening, the accumulation in stacks is the only procedure so far practiced by several countries as a means to get rid of this industrial externality. As part of a NATO Science for Peace Project (SfP 983311) this study describes the application of an environmental risk assessment (ERA) framework, to assess the risks posed by a PG stack to the surrounding soils, in Sfax, Republic of Tunisia. The ERA followed a weight of evidence approach, supported by two lines of evidence (LoE): the chemical (ChemLoE) and the ecotoxicological (EcotoxLoE). Integrated risks point for risk values greater than 0.5 in soils collected in PG stack surrounding area. Soil salinization, has likely contributed to the exacerbation of risks, as well as to the lack of consistency between both LoEs. This study highlights the need of rethinking the weight given to each LoE in ERA, in areas where soil salinization is a reality., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier Ltd.)

For regulatory purposes, the concentration addition model is the default first tier for assessing joint-action toxicity of metal mixtures. Although many researchers have evaluated binary and ternary mixtures, fewer have investigated joint-action toxicity in more complex mixtures, where deviations from additivity are more likely due to the greater number of potential interactions. In this study, we tested fixed ratios of five metals (lead, copper, nickel, zinc, cobalt) as metal oxide mixtures on three soil invertebrate species (Enchytraeus crypticus, Folsomia candida, Oppia nitens) at different dose effect levels (EC10-EC90) in an acid sandy forest and a loamy soil. Total metal concentrations for mixture ratios in soil did not explain or correlate with species responses. For F. candida, toxicity was linked to metal solubility, while for O. nitens and E. crypticus, toxicity did not correlate with total or extractable metals. In O. nitens and E. crypticus, however, soil ingestion could be an important route of exposure. Analysing the joint effect of metal mixtures, F. candida response was globally additive, while E. crypticus and O. nitens both presented synergistic effects at low-dose effect levels. Estimations at the EC50 level underestimated the deviations from additivity which were larger at higher and especially lower effect levels. Testing across different effect concentrations (EC10-EC90) was an important tool allowing the identification of these larger deviations from additivity outside the EC50 threshold. Considering most protection thresholds are set below the EC50 level, and it was in this low effect range where the highest synergisms were observed, risk assessment schemes should test additivity at the target protection level using representative test organisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Wastes generated in pig production are widely used as agricultural fertilizers. Nevertheless, such form of disposal supplies large amounts of waste in soils annually and can cause environmental pollution. The ecological risk of this practice to soil organisms has received little attention. Ecotoxicological tests are used to evaluate the toxicity of contaminants added to the soil biota. The aimed to evaluate the effect of liquid pig manure (LPM) on the reproduction of Eisenia andrei and Enchytraeus crypticus when applied in natural soils. LPM doses caused effects on earthworm reproduction in both soils, with EC 50 of 112 and 150 m 3  ha -1 in the Entisol and Nitosol, respectively. On enchytraeids, LPM had bigger effects, leading to EC 50 of 17.7 and 45.0 m 3  ha -1 in the Entisol and Nitosol, respectively. The results emphasize the importance to consider the ecological risks of LPM of conducting studies with natural soils and edaphic fauna as indicators.

Hunting activity is usually seen as a factor capable of causing an intense stress response in wildlife that may lead to short but also long-term stress. In the Lousã Mountain, Portugal, the population of red deer ( Cervus elaphus ) is the target of intensive seasonal hunting. We collected and measured cortisol (and its metabolites) in three tissues types (blood, feces and hair) from red deer hunted during two hunting seasons to evaluate the stress levels at different time windows. We also assessed the immunological and physical condition of the animals. We predicted that the hunting activity would act as a stressor inducing increased short and long-term stress levels in the population. Results showed an increase in hair cortisol levels during the months of harvesting. Surprisingly, the tendency for plasma cortisol levels was to decrease during the hunting season, which could be interpreted as habituation to hunting activity, or due to the hunting duration. Contrary to our predictions, fecal cortisol metabolites did not show any clear patterns across the months. Overall, our results suggest an influence of hunting activities on the physiological stress in red deer. In addition, hair seems to be useful to measure physiological stress, although more studies are required to fully understand its suitability as an indicator of long-term stress. Methodologically, our approach highlights the importance of simultaneously using different methods to assess short and long-term effects in studies on physiological stress reactions., Competing Interests: The authors declare no conflicts of interest.

Projected climate change and rainfall variability will affect soil microbial communities, biogeochemical cycling and agriculture. Nitrogen (N) is the most limiting nutrient in agroecosystems and its cycling and availability is highly dependent on microbial driven processes. In agroecosystems, hydrolysis of organic nitrogen (N) is an important step in controlling soil N availability. We analyzed the effect of management (ecological intensive vs. conventional intensive) on N-cycling processes and involved microbial communities under climate change-induced rain regimes. Terrestrial model ecosystems originating from agroecosystems across Europe were subjected to four different rain regimes for 263 days. Using structural equation modelling we identified direct impacts of rain regimes on N-cycling processes, whereas N-related microbial communities were more resistant. In addition to rain regimes, management indirectly affected N-cycling processes via modifications of N-related microbial community composition. Ecological intensive management promoted a beneficial N-related microbial community composition involved in N-cycling processes under climate change-induced rain regimes. Exploratory analyses identified phosphorus-associated litter properties as possible drivers for the observed management effects on N-related microbial community composition. This work provides novel insights into mechanisms controlling agro-ecosystem functioning under climate change.

Arsenic (As) is toxic to humans and the environment. Its toxicity has been assessed in many ways, including plant growth tests integrated in As risk assessments (RA). The tiered approach used in RA schemes assumes that lower tiers are more conservative than higher tiers. Plant growth tests may comprise lower tier of a RA and include the measurement of several endpoints. However, only few of these endpoints are highly sensitive and reliable, which makes them more appropriate to comprise lower tiers. Therefore, the selection of those endpoints is needed. The present study aimed to evaluate the most appropriate endpoints of plant growth tests to use in lower tier As RA schemes. This selection of endpoints was based on their sensitivity and reliability, using different tropical soils and plant species. In order to achieve this objective, six plant species were exposed to eight levels of As contamination (0; 8; 14.5; 26; 46.5; 84; 150; 270 mg kg -1 ), in three different tropical soils (Oxisol, Inceptisol, and tropical artificial soil). The endpoints measured were: first germination count (FrC), plant height (PH), relative leaf area (RLA), stem diameter (SD), total germination (TG), germination speed index (GSI), dry mass (DM), number of completely expanded leaves (CEL), plant survival (PS), soil plant analysis development chlorophyll level (SPAD), and the final germination count (FnC). Toxic values for 50% of effect were estimated for each endpoint within each species and test soil, to rank them according to their sensitivity and reliability. The most sensitive endpoints were: FrC, RLA, DM, GSI, PH, and FnC, while the most reliable endpoints were: FrC, DM, GSI, DM, PH, FnC, and TG. Our findings suggest that FrC, DM, GSI, PH, and FnC are the most adequate endpoints to be used in plant growth tests as lower tiers of As RA in tropical regions., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In soil metal ecotoxicology research, dosing is usually performed with metal salts, followed by leaching to remove excess salinity. This process also removes some metals, affecting metal mixture ratios as different metals are removed by leaching at different rates. Consequently, alternative dosing methods must be considered for fixed ratio metal mixture research. In this study three different metal mixture dosing methods (nitrate, oxide and annealed metal dosing) were examined for metal concentrations and toxicity. In the nitrate metal dosing method leaching reduced total metal retention and was affected by soil pH and cation exchange capacity (CEC). Acidic soils 3.22 (pH 3.4, CEC 8 meq/100g) and WTRS (pH 4.6, CEC 16 meq/100g) lost more than 75 and 64% of their total metals to leaching respectively while Elora (6.7 pH, CEC 21 meq/100g) and KUBC (pH 5.6, CEC 28 meq/100g) with higher pH and CEC only lost 13.6% and 12.2% total metals respectively. Metal losses were highest for Ni, Zn and Co (46.0%, 63.7% and 48.4% metal loss respectively) whereas Pb and Cu (5.6% and 20.0% metal loss respectively) were mostly retained, affecting mixture ratios. Comparatively, oxide and annealed metal dosing which do not require leaching had higher total metal concentrations, closer to nominal doses and maintained better mixture ratios (percent of nominal concentrations for the oxide metal dosing were Pb = 109.9%, Cu = 84.6%, Ni = 101.9%, Zn = 82.3% and Co = 97.8% and for the annealed metal dosing were Pb = 81.7%, Cu = 80.3%, Ni = 100.5%, Zn = 89.2% and Co = 101.3%). Relative to their total metal concentrations, nitrate metal dosing (lowest metal concentrations) was the most toxic followed by metal oxides dosing while the annealed dosing method was generally non-toxic. Due to the lack of toxicity of the annealed metals and their higher dosing effort, metal oxides, are the most appropriate of the tested dosing methods, for fixed-ratio metal mixtures studies with soil invertebrates., Competing Interests: The authors have declared that no competing interests exist.

The recently approved European Union (EU) Circular Economy Package intends to boost the production of fertilizing products, such as composts obtained from urban wastes (UWC) and the harmonization of their quality standards (certification), to avoid market rejection. UWC quality assessment frameworks in Europe, including the Portuguese and EU latest proposal on regulation of UWC production/commercialization are mostly based on physical-chemical and agronomical characterization. These do not provide any insight on the fraction of contaminant/mixture of contaminants bioavailable for non-target organisms, nor the existence of potential antagonistic and/or synergistic effects on them. The main objective of the present work if to evaluate the effects of UWC application on crop soils using seven standard ecotoxicological tests. Five UWC, two derived from source-separated organic wastes and three from mixed urban wastes were selected and tested using a battery of ISO guideline assays with plants and soil invertebrates. The tested doses intended to simulate over-use and repeated application scenarios, common practices among farmers. The results showed that the highest toxicity was observed for the UWC originated from source-separated organic wastes, when using concentrations slightly above the maximum annual doses. Excepting for this UWC, all the derived NOEC (No Observed Effect Concentration) were equal or higher than the maximum annual doses. The UWC toxicity for the tested species increased as follows: T. aestivum < L. sativa < E. crypticus < F. candida < E. andrei. UWC salinity, rather than the content of potentially toxic elements (PTEs), could explain the negative effects observed, considering that the composts are all equally stabilized. These results reinforce the need to include data from biological susceptibility of the receptors at risk on the existing regulation, to obtain a more realistic view of the potential risks and to adapt the UWC application practices, ultimately boosting the confidence of target-consumers., (Copyright © 2019 Elsevier B.V. All rights reserved.)

One of the entry routes of arsenic (As) into the food chain is through the consumption of edible parts of crops contaminated by this element. Different plant species present distinctive As accumulation and tolerance capacities. These differences are also influenced by As availability and speciation in soils. This study assessed the effect of As contamination on plant emergence and initial growth, as well as on accumulated As contents in different crops grown in tropical soils. In addition, it was intended to verify the protection level of the current soil As prevention value adopted in Brazil, which should be applicable for conceivably other tropical soils in Latin America. Plants of maize, rice, sorghum, common bean, sunflower, and radish were cultivated in two different tropical soils (Oxisol and Inceptisol) and in a standard substrate (tropical artificial soil - TAS) dosed with As (0; 8; 14.5; 26; 46.5; 84; 150; 270 mg kg -1 ). Early germination, total dry mass, As content, and bioconcentration factor were evaluated. The EC 20 and EC 50 values (the As concentration for 20% or 50% of effect relative to control treatment) based on total As concentration were more variable among different soils than the corresponding EC 20 and EC 50 values based on extractable (phytoavailable) As concentration. From the studied species, common bean was the most sensitive and maize was the least sensitive to As. Those species were the ones that accumulated the lowest As levels in shoot tissues. Arsenic concentrations measured in plant tissues and estimated bioaccumulation factors were not related to relative As toxicity among species. Data obtained suggest that the current Brazilian prevention value for arsenic is adequate for soils with high arsenic adsorption capacity., (Copyright © 2019. Published by Elsevier B.V.)

The aims of this work was to investigate, in soil microcosms, the effects on soil microbial community structure and function of increasing concentrations of 4-Nonylphenol (NP). The lasts is a product of degradation of NPEOs (Nonylphenol polyethoxylates) with a known toxic and estrogenic capacity able to disrupt animal's hormonal systems. The effect of increasing concentrations of NP (0, 10, 30, 90, and 270 mg NP kg -1 of dry soil) in soil microcosms in three sampling dates (28, 56, and 112 days) over soil microbial activity and function were assessed. Soil microbial activity was estimated by microbial ATP content, and both bacterial and fungal communities composition were estimated using the terminal restriction fragment length polymorphism technique (T-RFLP). Abundance of ammonia-oxidizing bacteria (AOB) was estimated by qPCR of gene encoding for the bacterial ammonia-monoxygenase (amoA). Changes in biologically mediated soil properties were also assessed, namely water-soluble NH + 4 , NO - 2 and NO - 3 content, the two last allowing the assessment of mineralization rates. NP-spiking had some unexpected impacts on microbial community structure and functions, since (i) impacted both bacterial and fungal communities structure at the highest NP concentration tested, bacterial communities were resistant to lower concentrations, while fungal communities were increasingly impacted until the end of the incubation at day 112; (ii) no community structure resilience was observed in bacteria at the highest NP concentration nor for fungi at any concentration; (iii) microbial activity decreased with NP after 28 and 56 d, but increased in the last sampling at the highest concentrations tests, coupled to an enrichment in AOB taxa after 56 and 112 days, that at least partly explain also explain the observed speed up of nitrification rates., (Copyright © 2019 Elsevier Inc. All rights reserved.)

There is a growing conservation concern about the possible consequences of environmental contamination in the health of bat communities. Most studies on the effects of contaminants in bats have been focused on organic contaminants, and the consequences of bat exposure to metals and metalloids remain largely unknown. The aim of this study was to evaluate the suitability of external biological matrices (fur and wing membrane) for the assessment of exposure and bioaccumulation of metals in bats. The concentration of arsenic, cadmium, cobalt, chromium, copper, manganese, nickel, lead, selenium and zinc was measured in internal organs (liver, heart, brain), internal (bone) and external tissues (wing membrane, fur) collected from bat carcasses of four species (Hypsugo savii, Nyctalus leisleri, Pipistrellus pipistrellus, Pipistrellus pygmaeus) obtained in windfarm mortality searches. With the exception of zinc (P = 0.223), the results showed significant differences between the concentrations of metals in the analyzed tissues for all metals (P < 0.05). Significant differences were also found between organs/tissues (P < 0.001), metals (P < 0.001) and a significant interaction between organs/tissues and metals was found (P < 0.001). Despite these results, the patterns in terms of metal accumulation were similar for all samples. Depending on the metal, the organ/tissue that showed the highest concentrations varied, but fur and wing had the highest concentrations for most metals. The variability obtained in terms of metal concentrations in different tissues highlights the need to define standardized methods capable of being applied in monitoring bat populations worldwide. The results indicate that wing membrane and fur, biological matrices that may be collected from living bats, yield reliable results and may be useful for studies on bats ecotoxicology, coupled to a standardized protocol for large-scale investigation of metal accumulation., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The use of organic wastes as soil amendments can be an important measure to improve soil quality and reduce waste accumulation and landfilling. However, the potential contaminant loads of such wastes, can be a source of environmental concern. Consequently, legislation has been developed to regulate the use of these wastes in agricultural soils. However, the regulations only consider chemical parameters, which are insufficient to establish the level of environmental risk. A possible solution is the use of species sensitivity distributions (SSDs), employing ecotoxicological data from test batteries that could be incorporated into legislation. In the present study, 2 different hazardous concentrations affecting 5 and 50% of the soil community (HC5 and HC50, respectively) were determined using ecotoxicological data (effect concentrations, 10 and 50% [EC10 and EC50, respectively]) for 5 different wastes. The results demonstrate that, as expected, current legislative thresholds do not translate to environmental risk/protection and that SSDs may be an important tool allowing the simple inclusion and interpretation of ecotoxicological data from test batteries in legislation. On the other hand, SSDs must be used with caution because there are still doubts about their actual value in risk prediction and about which estimates provide adequate protection. For instance, the use of HC50 EC10 values is not recommended; these values overlap with the more conservative HC5 EC50 data, highlighting the fact that the use of lower effect concentrations may not always provide the most protective approach. Also, hazardous concentrations need to be calibrated at the field or semifield level, to verify environmental protection in different soils/environments and the adequacy of standard test organisms. Environ Toxicol Chem 2019;38:1569-1576. © 2019 SETAC., (© 2019 SETAC.)

Inadequate application of liquid pig manure (LPM) may pose risks to the soil due to the potential contaminants that exists, as well as by the large water input that can originate excessive moisture. By using Terrestrial Model Ecosystems, this study aimed to evaluate the effect of application of LPM (82% moisture) using the application rates of 20, 50 and 150 m 3 ha -1 and also of the corresponding amount of water to understand the origin of effects on the soil fauna of two sub-tropical soils. In general, the results obtained for the two soil types indicated that LPM (150 m 3 ha -1 ) changed the composition of soil fauna, with an increase in the abundance of insect larvae and dipterans, but a decrease in the number of earthworms and enchytraeids. Microbial biomass, soil respiration and the nutrients Mg, K and P increased with the amount of LPM supplied to the soil. When analysing the effects of adding large volumes of water via the LPM, results showed that application did not originate significant effects on the parameters measured. The differences obtained when comparing both treatments were mainly attributed to the organic load brought by the LPM and not to the amount of water without LPM., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The laboratory reproduction test with the predatory mite Hypoaspis aculeifer is currently a mandatory test in the new EU data requirements for prospective environmental risk assessment of Plant Protection Products (PPPs). However, the low sensitivity often shown by this mite towards PPPs, when compared to other invertebrates (namely Folsomia candida and Eisenia fetida), makes the test with this species not very useful in the lower tier test battery. However, the current test protocol only considers exposure to contaminants via contaminated soil, disregarding exposure via contaminated food and does not take into account the fact that H. aculeifer is a predatory species. Therefore, through this protocol, the toxicity of contaminants to soil mites might be underestimated and, thus, an adaptation of the test performance, by including exposure via contaminated food, may be necessary. With this aim, two reproduction tests with H. aculeifer were performed using copper chloride as model substance, artificial soil as test substrate and cheese mites as food. The OECD guideline was followed but, while in one test cheese mites from normal laboratory breeding cultures (clean prey mites) were provided, in the other test, cheese mites previously exposed to copper (Cu pre-exposed prey mites) were provided. Predatory mites were affected at lower concentrations in tests using Cu pre-exposed prey compared to test with clean-prey (NOEC = 1225 and 1508 mg kg -1 and EC 10  = 1204 and 1903 mg kg -1 using Cu pre-exposed and clean prey, respectively). However, this higher sensitivity was not detected by EC 50 values (EC 50  = 2634 and 2814 mg kg -1 using Cu pre-exposed and clean prey, respectively). Further tests are needed in order to (i) investigate the relevance of oral exposure to different PPPs, (ii) optimize the contamination of prey mites according to the chemical properties of each substance and (iii) substantiate a proposal to adapt the standard protocol., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Veterinary medicines are widely applied for the treatment and prevention of animal diseases. Consequently, animal manure contains significant amounts of environmental pollutants that are potential sources of environmental pollution when inappropriately applied in soils. This work aimed to evaluate ecotoxicological effects of doses of commercial fipronil, neem cake and neem extract in the survival and reproduction of earthworms (Eisenia andrei), enchytraeidae (Enchytraeus crypticus) and springtails (Folsomia candida) in Oxisol and tropical artificial soil (TAS). Applications of fipronil, neem cake and extract in soil were carried out according to standardized ISO methodologies by using a random experimental design with five replicates. Toxic effects of fipronil for springtails in Oxisol and TAS were observed with LC 50 of 0.26 mg kg -1 (0.18-0.35 mg kg -1 ) and 0.29 mg kg -1 (0.22-0.37 mg kg -1 ), respectively. It was not observed significant toxic effects of fipronil for earthworms and enchytraeidae in both soils. However, significant amounts of juvenile earthworm and adult enchytraeidae decreased in fipronil doses higher than 10 mg kg -1 . Neem cake and extract were not toxic for earthworms and enchytraeidae but, significant amounts of juvenile springtails decreased in neem cake doses from 500 to 1000 mg kg -1 . It can be concluded that the use of veterinary medicines containing synthetic compounds for preventing diseases in animals needs to be controlled to avoid environmental pollution after applying manure in soil. Veterinary medicines containing natural compounds as neem cake and extract are eco-friendly and could be efficiently applied in soil in a sustainable way., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The aim of this study is to evaluate the toxicity of the fipronil and its metabolites, fipronil sulfone and fipronil desulfinyl on Folsomia candida. Three types of soils were used to perform the ecotoxicological tests, two natural soils (Oxisol and Entisol), and an artificial substrate (Tropical Artificial Soil). The treatments consisted of increasing doses of a veterinary medicinal product with active ingredient based on fipronil, and of its two main metabolites, fipronil sulfone and fipronil desulfinyl. The doses of fipronil were 0.00, 0.02, 0.04, 0.08, 0.15, 0.30, 0.60, and 1.00 mg kg -1 of the active ingredient. For fipronil sulfone and fipronil desulfinyl the doses tested were: 0.00, 0.01, 0.02, 0.04, 0.06, 0.08, 0.15, 0.30, 0.60, 1.00, and 5.00 mg kg -1 . The effects of toxicity on F. candida were evaluated by survival and reproduction tests, based on the methods described by ISO 11267. There was a completely randomized design, with six replicates for the fipronil tests and eight for the metabolites. We show that fipronil and its metabolites, even at very low concentrations, are toxic to F. candida. Reduction of reproduction can be observed at concentrations of 0.15 mg kg -1 . There was little difference in toxicity (LC 50 and EC 50 ) between fipronil and its metabolites. Rather, differences were related more to soil type, where the artificial soil was more sensitive than the two natural soils (Oxisol and Entisol). We conclude that it is important to perform ecotoxicological tests in natural soils, in order to generate more realistic representations of veterinary drug toxicity on the environment., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Arbuscular mycorrhizal fungi (AMF) are mutualistic symbionts considered a key group in soil systems involved in the provision of several ecosystem services. Recently they have been listed by EFSA as organisms to be included in the test battery for the risk assessment of plant protection product (PPPs). This study aimed to contribute to improve the ISO Protocol (ISO 10832: 2009) by assessing the feasibility of using other AMF species under different test conditions. Overall, results showed that AMF species Gigaspora albida and Rhizophagus clarus (selected out of five AMF species) are suitable to be used in spore germination tests using the ISO protocol (14 days incubation with sand or artificial soil as substrate) to test PPPs. However, several modifications to the protocol were made in order to accommodate the use of the tested isolates, namely the incubation temperature (28 °C instead of 24 °C) and the change of reference substance (boric acid instead of cadmium nitrate). The need for these changes, plus the results obtained with the three fungicides tested (chlorothalonil, mancozeb and metalaxyl-M) and comparisons made with literature on the relevance of the origin of AMF isolates in dictating the adequate test conditions, emphasize the importance of adjusting test conditions (AMF species/isolates and test temperature) when assessing effects for prospective risk assessment targeting different climatic zones. So, further studies should be conducted with different AMF species and isolates from different climatic regions, in order to better define which species/isolate and test conditions should be used to assess effects of a particular PPP targeting a given climatic zone.

Increasing climatic and anthropogenic pressures on soil ecosystems are expected to create a global patchwork of disturbance scenarios. Some regions will be strongly impacted by climate change, others by agricultural intensification, and others by both. Soil microbial communities are integral components of terrestrial ecosystems, but their responses to multiple perturbations are poorly understood. Here, we exposed soils from sustainably- or intensively-managed grasslands in an agro-silvo-pastoral oak woodland to month-long intensified drought and flood simulation treatments in a controlled mesocosm setting. We monitored the response of the bacterial communities at the end of one month as well as during the following month of recovery. The communities in sustainably-managed plots under all precipitation regimes were richer and more diverse than those in intensively-managed plots, and contained a lower proportion of rapidly-growing taxa. Soils from both land managements exhibited changes in bacterial community composition in response to flooding, but only intensively-managed soils were affected by drought. The ecologies of bacteria favored by both drought and flood point to both opportunism and stress tolerance as key traits shaping the community following disturbance. Finally, the response of several taxa (i.e. Chloracidobacteria RB41, Janthinobacterium sp.) to precipitation depended on land management, suggesting that the community itself affected individual disturbance responses. Our findings provide an in-depth view of the complexity of soil bacterial community responses to climatic and anthropogenic pressures in time, and highlight the potential of these stressors to have multiplicative effects on the soil biota., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Neonicotinoids are the most prominent group of insecticides in the world and are commercialized in over 120 countries for the control of agricultural pests mainly due to their broad-spectrum activity and versatility in application. Though non-target soil organisms are likely to be exposed during application, there is paucity of information in scientific literature regarding their sensitivity to neonicotinoids. This study attempts to fill this gap by evaluating, under laboratory conditions, the chronic toxicity of the neonicotinoids thiacloprid and acetamiprid, through their commercial formulations (CF), to the soil invertebrates Folsomia candida, Eisenia andrei and Enchytraeus crypticus. Results obtained indicate that the relative reproductive sensitivity of the test organisms can be expressed as: F. candida = E. andrei > E. crypticus (for acetamiprid CF) and E. andrei > F. candida > E. crypticus (for thiacloprid CF). To extrapolate from laboratory test results to field conditions, predicted environmental concentrations (PECs) and predicted no-effect concentrations were derived. Calculated toxicity-exposure ratios (TER = EC10/PEC) were below trigger values for acetamiprid and thiacloprid, when estimated with initial PEC. While estimated hazard quotients (HQ = PEC/PNEC), were greater than the European Commission trigger value. Therefore, with the current data under standard environmental risk assessment schemes it can be considered that the risk of thiacloprid and acetamiprid to the soil compartment is unacceptable. However, further research into the effects of these substances on different organisms is required to increase the confidence in the risk assessment estimates for instance, by calculating hazardous concentrations using species sensitivity distribution curves., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Soils from some Latin American countries support the highest biodiversity levels on the planet and simultaneously have some of the most serious environmental impacts attributed to both historical and current agricultural practices and industrial activities. Soil contamination has resulted from intensive use of pesticides, extensive mining and other industrial activities, and uncontrolled management of waste within inappropriate regulatory frameworks. The present study presents an overview of the scientific research on soil ecotoxicology conducted in Latin America, summarizing the recent advances and highlighting the needs for further refinements in this research field. Most of the contributions to the scientific literature have been from Brazil. The most investigated issue is the ecotoxicity of pesticides and earthworms, which were the organisms most frequently used as test species. Needs identified by Latin American researchers include methods and procedures for: 1) identifying and collecting natural soils to be used as reference test-substrates in tests, 2) identifying and discerning the range of sensitivities of native test species to soil contaminants, 3) developing environmental guidelines applicable to tropical/subtropical conditions, and 4) developing methods and procedures for higher tier testing for full development and implementation of environmental risk assessment schemes. The protection of Latin American soils, including provision of goods and services, is currently framed in legislation and other regulations, but implementation requires significant improvement and additional training programs. Environ Toxicol Chem 2017;36:1795-1810. © 2017 SETAC., (© 2017 SETAC.)

Using organic wastes, as soil amendments, is an important alternative to landfilling with benefits to soil structure, water retention, soil nutrient and organic matter concentrations. However, this practice should be monitored for its environmental risk due to the frequent presence, of noxious substances to soil organisms. To evaluate the potential of eight organic wastes with different origins, as soil amendments, reproduction tests with four soil invertebrate species (Folsomia candida, Enchytraeus crypticus, Hypoaspis aculeifer, Eisenia fetida) were performed using gradients of soil-waste mixtures. Results obtained demonstrated that contaminant concentrations required by current legislation might not be a protective measure for the soil ecosystem, as they do not properly translate the potential toxicity of wastes to soil invertebrates. Some wastes with contaminant loadings below thresholds showed higher toxicity than wastes with contaminants concentrations above legal limits. Also, test organism reproduction was differently sensitive to the selected wastes, which highlights the need to account for different organism sensitivities and routes of exposure when evaluating the toxicity of such complex mixtures. Finally this study shows that when combining chemical and ecotoxicological data, it is possible to postulate on potential sources of toxicity, contributing to better waste management practices and safer soil organic amendment products., (Copyright © 2017 Elsevier B.V. All rights reserved.)