Search

International audience; Preservation of biodiversity and ecosystem services is critical for sustainable development and human well-being. However, an unprecedented erosion of biodiversity is observed and the use of plant protection products (PPP) has been identified as one of its main causes. In this context, at the request of the French Ministries responsible for the Environment, for Agriculture and for Research, a panel of 46 scientific experts ran a nearly 2-year-long (2020-2022) collective scientific assessment (CSA) of international scientific knowledge relating to the impacts of PPP on biodiversity and ecosystem services. The scope of this CSA covered the terrestrial, atmospheric, freshwater, and marine environments (with the exception of groundwater) in their continuity from the site of PPP application to the ocean, in France and French overseas territories, based on international knowledge produced on or transposable to this type of context (climate, PPP used, biodiversity present, etc.). Here, we provide a brief summary of the CSA's main conclusions, which were drawn from about 4500 international publications. Our analysis finds that PPP contaminate all environmental matrices, including biota, and cause direct and indirect ecotoxicological effects that unequivocally contribute to the decline of certain biological groups and alter certain ecosystem functions and services. Levers for action to limit PPP-driven pollution and effects on environmental compartments include local measures from plot to landscape scales and regulatory improvements. However, there are still significant gaps in knowledge regarding environmental contamination by PPPs and its effect on biodiversity and ecosystem functions and services. Perspectives and research needs are proposed to address these gaps.

A wide diversity of plant protection products (PPP) is used for crop protection leading to the contamination of soil, water, and air, which can have ecotoxicological impacts on living organisms. It is inconceivable to study the effects of each compound on each species from each compartment, experimental studies being time consuming and cost prohibitive, and animal testing having to be avoided. Therefore, numerous models are developed to assess PPP ecotoxicological effects. Our objective was to provide an overview of the modeling approaches enabling the assessment of PPP effects (including biopesticides) on the biota. Six categories of models were inventoried: (Q)SAR, DR and TKTD, population, multi-species, landscape, and mixture models. They were developed for various species (terrestrial and aquatic vertebrates and invertebrates, primary producers, micro-organisms) belonging to diverse environmental compartments, to address different goals (e.g., species sensitivity or PPP bioaccumulation assessment, ecosystem services protection). Among them, mechanistic models are increasingly recognized by EFSA for PPP regulatory risk assessment but, to date, remain not considered in notified guidance documents. The strengths and limits of the reviewed models are discussed together with improvement avenues (multigenerational effects, multiple biotic and abiotic stressors). This review also underlines a lack of model testing by means of field data and of sensitivity and uncertainty analyses. Accurate and robust modeling of PPP effects and other stressors on living organisms, from their application in the field to their functional consequences on the ecosystems at different scales of time and space, would help going toward a more sustainable management of the environment. Graphical Abstract Combination of the keyword lists composing the first bibliographic query. Columns were joined together with the logical operator AND. All keyword lists are available in Supplementary Information at https://doi.org/10.5281/zenodo.5775038 (Larras et al. 2021).

While long-term organic fertilizer (OF) applications tend to decrease copper (Cu) and zinc (Zn) availability in agricultural soils, earthworm bioturbation has been reported to have the opposite effect. Thus, the consequences of OF amendments in earthworm-inhabited soils on Cu and Zn bioavailability to earthworms are still under debate. Here, we assessed the effect of a decade of agronomically realistic OF applications on Cu and Zn availability in earthworm-inhabited soils and the consequences on Cu and Zn bioavailability to earthworms. An epi-endogeic species (Dichogaster saliens) was exposed in microcosms to three field-collected soils that had received either no, mineral, or organic fertilization for a decade. Dissolved organic matter (DOM) properties (i.e., concentration, aromaticity, and binding properties toward Cu), pH, and Cu and Zn availability (i.e., total concentration and free ionic activity) were determined in the solution of the soil containing earthworms. Cu and Zn bioavailability was assessed by measuring the net accumulation (ng) and concentration of Cu and Zn in earthworms (mg kg−1). Despite soil Cu and Zn contamination induced by a decade of OF applications, organic fertilization induced an increase in soil pH and DOM properties that drove the reduction of Cu and Zn availability in earthworm-inhabited soils, while bioturbation had little effect on soil pH, DOM properties, and Cu and Zn availability. Consistently, Cu and Zn bioavailability to earthworms did not increase with OF applications. From an ecotoxicological perspective, our results suggest that agronomically realistic applications of OF for a decade should not pose a risk to earthworms in terms of Cu and Zn net accumulation, but further studies have to be undertaken to understand consequent long-term toxicity after exposure.

Pesticides may harm soil organisms such as earthworms and enchytraeids, but knowledge is lacking on their relative sensitivity to these chemicals and the consequences on soil functions. The aim of this study was to assess the impact of exposure to a commercial fungicide formulation (Swing® Gold, containing dimoxystrobin and epoxiconazole) on the function of earthworms (Aporrectodea caliginosa) and enchytraeids (Enchytraeus buchholzi) in soil organic matter (SOM) mineralization. The soil organisms were incubated alone and together in a 26-day laboratory experiment. At the recommended field rate, the fungicide induced a decrease in the SOM mineralization and a delay in the maximum daily CO2 emissions compared to the control soil without fungicide. Soil fauna also influenced SOM mineralization with a higher cumulated CO2 release after 26 days in the control soil with earthworms (by 21%) than without fauna. When both earthworms and enchytraeids were present, SOM mineralization did not increase, and there was a negative effect on earthworm weight gain. Finally, an alteration of fauna influence with treatment was observed from day 19, meaning that the effect of fauna on SOM mineralization changed with fungicide treatment. Earthworms no longer promoted SOM mineralization when fungicide was present at three-fold the recommended field rate. The effects of enchytraeids on SOM mineralization were similar with and without fungicide exposure. This study underlines the importance of considering the relative sensitivity of soil organisms to environmental factors and interactions between them when assessing soil functioning.

A wide diversity of plant protection products (PPP) is used for crop protection leading to the contamination of soil, water, and air, which can have ecotoxicological impacts on living organisms. It is inconceivable to study the effects of each compound on each species from each compartment, experimental studies being time consuming and cost prohibitive, and animal testing having to be avoided. Therefore, numerous models are developed to assess PPP ecotoxicological effects. Our objective was to provide an overview of the modeling approaches enabling the assessment of PPP effects (including biopesticides) on the biota. Six categories ofmodels were inventoried: (Q)SAR, DR and TKTD, population, multi-species, landscape, and mixture models. They were developed for various species (terrestrial and aquatic vertebrates and invertebrates, primary producers, micro-organisms) belonging to diverse environmental compartments, to address different goals (e.g., species sensitivity or PPP bioaccumulation assessment, ecosystem services protection). Among them, mechanistic models are increasingly recognized by EFSA for PPP regulatory risk assessment but, to date, remain not considered in notified guidance documents. The strengths and limits of the reviewed models are discussed together with improvement avenues (multigenerational effects, multiple biotic and abiotic stressors). This review also underlines a lack of model testing by means of field data and of sensitivity and uncertainty analyses. Accurate and robust modeling of PPP effects and other stressors on living organisms, from their application in the field to their functional consequences on the ecosystems at different scales of time and space, would help going toward a more sustainable management of the environment.

Before plant protection product (PPP) marketing authorization, a risk assessment for nontarget soil organisms (e.g., earthworms) is required as part of Regulation (EC) No. 1107/2009. Following a stepwise approach, higher tier earthworm field studies are needed if they cannot demonstrate low long-term risk based on laboratory studies. The European guidance for terrestrial ecotoxicology refers to ISO guideline 11268-3 as a standard to conduct earthworm field studies. Assessment of such studies may be challenging, as no European harmonized guidance is available to properly analyze the accuracy, representativeness, and appropriateness of experimental designs, as well as the statistical analysis robustness of results and their scientific reliability. Following the ISO guideline 11268-3, a field study was performed in 2016-2017 (Versailles, France). An assessment of the first year of this field study was performed in agreement with the quality criteria provided in 2006 in the guidance document published by de Jong and collaborators and recommendations by Kula and collaborators that allows describing the protocol and results of earthworm field studies. Not only did we underline the importance of a detailed analysis of raw data on the effects of pesticides on earthworms in field situations, but we also provided recommendations to harmonize protocols for assessing higher tier field studies devoted to earthworms to advance a better assessment of PPP fate and ecotoxicity. In particular, we provided practical field observations related to the study design, pesticide applications, and earthworm sampling. Concurrently, in addition to the conventional earthworm community study, we propose carrying out an assessment of soil function (i.e., organic matter decomposition, soil structuration, etc.) and calculating diversity indices to obtain information about earthworm community dynamics after the application of PPPs. Finally, through field observations, any relevant observation of external and/or internal recovery should be reported. Integr Environ Assess Manag 2023;19:254-271. © 2022 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental ToxicologyChemistry (SETAC).

International audience; The pesticide risk assessment for earthworms is currently performed using standardized tests, the model species Eisenia fetida, and the analyses of the data obtained are performed with ad hoc statistical tools. We assessed the impact of two fungicides on the entire growth pattern of the earthworm species Aporrectodea caliginosa, which is highly representative of agricultural fields. Individuals of three different ages (from hatching to 56 days old) were exposed to Cuprafor micro(R) (copper oxychloride) and Swing(R) Gold (dimoxystrobin and epoxiconazole). Data were analyzed with an energy-based toxicodynamic model coupled with a toxicokinetic model. The copper fungicide caused a drastic growth inhibition once the no effect concentration (NEC), estimated at 65 mg kg(-1) of copper, was exceeded. The Swing(R) Gold negatively affected the growth with NEC values estimated at 0.387 mg kg(-1) and 0.128 mg kg(-1) for the dimoxystrobin and the epoxiconazole in this fungicide formulation, respectively. The time-profile of the effects on A. caliginosa individuals was fully accounted for by the model, whatever their age of exposure. Furthermore, toxicity data analyses, supported by measurements of fungicide concentrations in earthworm at the end of the experiment, allowed bettering understanding of the mechanisms of action of the fungicides towards earthworm growth.

It is essential to monitor pesticides in soils as their presence at trace levels and their bioavailability can induce adverse effects on soil's ecosystems, animals, and human health. In this study, we developed an analytical method for the quantification of traces of multi-class pesticides in soil using liquid chromatography-tandem mass spectrometry. In this way, 31 pesticides were selected, including 12 herbicides, 9 insecticides, and 10 fungicides. Two extraction techniques were first evaluated, namely, the pressurized liquid extraction and the QuEChERS procedure. The latest one was finally selected and optimized, allowing extraction recoveries of 55 to 118%. The role of the chelating agent EDTA, which binds preferentially to soil cations that complex some pesticides, was highlighted. Coupled with liquid chromatography-tandem mass spectrometry, the procedure displayed very high sensitivity, with limits of quantification (LOQ) in the range 0.01-5.5 ng/g. A good linearity (R

The use of pesticides in agriculture to protect crops against pests and diseases generates environmental contamination. The atmospheric compartment contributes to their dispersion at different distances from the application areas and to the exposure of organisms in untreated areas through dry and wet deposition. A multiresidue analytical method using the same TD-GC-MS analytical pipeline to quantify pesticide concentrations in both the atmosphere and rainwater was developed and tested in natura. A Box-Behnken experimental design was used to identify the best compromise in extraction conditions for all 27 of the targeted molecules in rainwater. Extraction yields were above 80% except for the pyrethroid family, for which the recovery yields were around 40-59%. TD-GC-MS proved to be a good analytical solution to detect and quantify pesticides in both target matrices with low limits of quantification. Twelve pesticides (six fungicides, five herbicides and one insecticide) were quantified in rainwater at concentrations ranging from 0.5 ng·L

International audience; A new sensitive and selective analytical methodology to quantify glyphosate (GLY), aminomethylphosphonic acid (AMPA), and glufosinate (GLU) in both soil and earthworms (Allolobophora chlorotica) was developed. The extraction and purification methods were optimized. The samples were extracted with various aqueous solutions (HNO3, H2O, KOH and borate buffer) and derivatized with 9-Fluorenylmethyl chloroformate (FMOCCl). To optimize the extraction step, a method to remove the excess FMOCCl was applied based on liquid-liquid extraction with diethyl ether. The purification of derivatized extracts was carried out using XLB solid phase extraction (SPE) cartridges before internal standard quantification by liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS). The elution step was optimized to obtain the best recoveries possible, which was with acidic methanol (1% formic acid) (67% for GLY, 70% for GLU and 65% for AMPA). The extraction and purification method followed by analysis of the two herbicides and AMPA in soils using LC/MS/MS determined limit of quantification (LOQ) values of 0.030mug g(-)(1) for GLY, 0.025mug g(-)(1) for AMPA and 0.020microg g(-)(1) for GLU . For earthworms, LOQ were 0.23mug g(-)(1) for GLY, 0.20mug g(-)(1) for AMPA and 0.12mug g(-)(1) for GLU. . The developed method was applied to determine these compounds in natural soils and earthworms.

International audience; Before their placing on the market, the safety of plant protection products (PPP) towards both human and animal health, and the environment has to be assessed using experimental and modelling approaches. Models are crucial tools for PPP risk assessment and some even help to avoid animal testing. This review investigated the use of modelling approaches in the ecotoxicology section of PPP active substance assessment reports prepared by the authorities and opened to consultation from 2011 to 2021 in the European Union. Seven categories of models (Structure-Activity, ToxicoKinetic, ToxicoKinetic-ToxicoDynamic, Species Sensitivity Distribution, population, community, and mixture) were searched for into the reports of 317 active substances. At least one model category was found for 44 % of the investigated active substances. The most detected models were Species Sensitivity Distribution, Structure-Activity and ToxicoKinetic for 27, 21 and 15 % of the active substances, respectively. The use of modelling was of particular importance for conventional active substances such as sulfonylurea or carbamates contrary to microorganisms and plant derived substances. This review also highlighted a strong imbalance in model usage among the biological groups considered in the European Regulation (EC) No 1107/2009. For example, models were more often used for aquatic than for terrestrial organisms (e.g., birds, mammals). Finally, a gap between the set of models used in reports and those existing in the literature was observed highlighting the need for the implementation of more sophisticated models into PPP regulation.

Massive use of pesticides in conventional agriculture leads to accumulation in soil of complex mixtures, triggering questions about their potential ecotoxicological risk. This study assessed cropland soils containing pesticide mixtures sampled from conventional and organic farming systems at La Cage and Mons, France. The conventional agricultural field soils contained more pesticide residues (11 and 17 versus 3 and 11, respectively) and at higher concentrations than soils from organic fields (mean 6.6 and 10.5 versus 0.2 and 0.6 μg kg−1, respectively), including systemic insecticides belonging to neonicotinoids, carbamate herbicides and broad-spectrum fungicides mostly from the azole family. A risk quotient (RQi) approach evaluated the toxicity of the pesticide mixtures in soil, assuming concentration addition. Based on measured concentrations, both conventional agricultural soils posed high risks to soil invertebrates, especially due to the presence of epoxiconazole and imidacloprid, whereas soils under organic farming showed negligible to medium risk. To confirm the outcome of the risk assessment, toxicity of the soils was determined in bioassays following standardized test guidelines with seven representative non-target invertebrates: earthworms (Eisenia andrei, Lumbricus rubellus, Aporrectodea caliginosa), enchytraeids (Enchytraeus crypticus), Collembola (Folsomia candida), oribatid mites (Oppia nitens), and snails (Cantareus aspersus). Collembola and enchytraeid survival and reproduction and land snail growth were significantly lower in soils from conventional compared to organic agriculture. The earthworms displayed different responses: L. rubellus showed higher mortality on soils from conventional agriculture and large body mass loss in all field soils, E. andrei showed considerable mass loss and strongly reduced reproduction, and A. caliginosa showed significantly reduced acetylcholinesterase activity in soils from conventional agriculture. The oribatid mites did not show consistent differences between organic and conventional farming soils. These results highlight that conventional agricultural practices pose a high risk for soil invertebrates and may threaten soil functionality, likely due to additive or synergistic “cocktail effects”.

The burrowing, feeding and foraging activities of terrestrial and benthic organisms induce displacements of soil and sediment materials, leading to a profound mixing of these media. Such particle movements, called "sediment reworking" in aquatic environments and "bioturbation" in soils, have been thoroughly studied and modeled in sediments, where they affect organic matter mineralization and contaminant fluxes. In comparison, studies characterizing the translocation, by soil burrowers, of mineral particles, organic matter and adsorbed contaminants are paradoxically fewer. Nevertheless, models borrowed from aquatic ecology are used to predict the impact of bioturbation on organic matter turnover and contaminant transport in the soil. However, these models are based on hypotheses that have not been tested with adequate observations in soils, and may not necessarily reflect the actual impact of soil burrowers on particle translocation. This paper aims to (i) highlight the possible shortcomings linked to the current use of sediment reworking models for soils, (ii) identify how recent progresses in aquatic ecology could help to circumvent these limitations, and (iii) propose key steps to ensure that soil bioturbation models are built on solid foundations: more accurate models of organic matter turnover, soil evolution and contaminant transport in the soil are at stake.

International audience; In a companion study, carried out in a fallow meadow close to a lead recycling factory, we showed that earthworms were absent in the first 20 m and then gradually increased in abundance from 30 to 110 m from the factory. Here we assessed in the same meadow whether these differences in earthworm abundance were associated with the loss of physical soil properties. Soil cores were sampled and infiltration measured in situ at five distances from the factory (10, 30, 50, 80 and 110 m). X-ray tomography was used to characterize the earthworm burrow systems within cores. The burrow systems were minimal at the first two distances, with the only macropores observed probably produced by insects such as ants. Typical earthworm burrows were seen at 50 m but most of their characteristics (volume, diameter, continuity) were similar to those observed at 10 or 30 m. Dense and well-developed burrow systems were observed at 80 and 110 m from the factory with significantly larger volume and continuity. Burrow diameter at 80 m was significantly higher than at closer distances but it significantly decreased at 110 m associated with the higher abundance of endogeics earthworm species. Water infiltration followed the same trend with significantly lower rates at the first two distances compared to those further from the factory, where rates increased from 70 to 250%. This study emphasizes the need to develop a more functional approach to study the spatial effects of contamination on soil ecological processes and fertility, beyond modifications in earthworm communities.

Interactions of earthworm species with their environment and in particular the soil they inhabit is poorly understood. In this study, we aimed to use mid infrared absorption (MIR) spectroscopy of earthworm bodies in combination with discriminant analysis of principal components (DAPC), in order to investigate if their biogeochemical composition is determined by their species belonging or the soil they inhabit. Our conceptual approach included the comparison of MIR spectra of four earthworm species (Lumbricus terrestris, Allolobophora chlorotica, Aporrectodea caliginosa and Aporrectodea icterica) sampled from four different agricultural fields in France. Moreover, we tested the influence of the habitat on the earthworm MIR spectra by exchanging individuals and soils in an incubation experiment. MIR spectra varied according to species and sampling site. The MIR spectra of earthworm bodies also changed during the incubation experiment, and for L. terrestris, A. chlorotica and A. icterica depended on the properties of the soil ingested. This suggested that the food source in different habitats impacted the biogeochemical composition of earthworm bodies. DAPC allowed for the identification of earthworm species ranging from 64 to 100% of accuracy (mean 78%). We thus conclude that MIR spectroscopy may be a suitable approach to identify earthworm species, but that their signature may also allow to investigate their ecology as determined by the soil they inhabit.

International audience; Critical knowledge gaps about environmental fate and unintentional effects of currently used pesticides (CUPs) hamper the understanding and mitigation of their global impacts on ecological processes. We investigated the exposure of earthworms to 31 multiclass CUPs in an arable landscape in France. We highlighted the presence of at least one pesticide in all soils (n=180) and 92% of earthworms (n=155) both in treated crops and nontreated habitats (hedgerows, grasslands, and cereals under organic farming). Mixtures of at least one insecticide, one herbicide, and one fungicide (> limit of quantification) contaminated 90% of soils and 54% of earthworms at levels that could endanger these nontarget beneficial soil organisms. A high risk of chronic toxicity to earthworms was found (46% of samples) both in treated winter cereals and nontreated habitats considered as refuges. This may alter biodiversity, hinder recovery, and impair ecosystem functions. These results provide essential insights for sustainable agriculture and CUP regulation, and highlight the potential of pesticides as agents of global change.

International audience; The representativeness of laboratory studies of the fate of pesticides in soil in field conditions is questionable. This study aimed at comparing the dissipation and bioavailability to earthworms of two fungicides, dimoxystrobin (DMX) and epoxiconazole (EPX), over 12 months under laboratory and field conditions. In both approaches, the fungicides were applied to the same loamy soil as a formulated mixture at several concentrations. We determined total DMX and EPX concentrations in the soil using exhaustive extraction, their environmental availability using mild extraction and their bioavailability through internal concentrations in exposed earthworms. The initial fungicide application appeared as much better controlled in terms of dose and homogeneity in the laboratory than in the field. One year after application, a similar dissipation rate was observed between the laboratory and field experiments (ca 80% and 60% for DMX and EPX, respectively). Similarly, the ratio of available/total concentrations in soil displayed the same trend whatever the duration and the conditions (field or lab), EPX being more available than DMX. Finally, the environmental bioavailability of the two fungicides to earthworms was heterogeneous in the field, but, in the laboratory, the bioaccumulation was evidenced to be dose-dependent only for DMX. Our findings suggest that the actual fate of the two considered fungicides in the environment is consistent with the one determined in the laboratory, although the conditions differed (e.g., presence of vegetation, endogeic earthworm species). This study allowed better understanding of the fate of the two considered active substances in the soil and underlined the need for more research dedicated to the link between environmental and toxicological bioavailability.

International audience; In the context of global changes, interest is growing in biodiversity in urban areas. However, sampling biodiversity in urbanized environments faces challenges. For soil organisms such as earthworms, sampling methods exist and have been tested in natural and agricultural areas, yet their efficiency and acceptability in urbanized areas has not been sufficiently addressed. This study compared the efficiency of two classical methods (with different environmental impact) for the quantitative assessment of earthworm populations in landscape contexts with varying levels of urbanization. For that, 15 sites near Paris (France) were sampled in autumn 2016 and spring 2017 with an electrical extraction method and a combined mustard oil liquid extraction followed by hand-sorting. Both methods produced similar estimates of species richness (number of sampled species) and juveline/adult ratios, but the electrical method extracted significantly fewer earthworms than the combined mustard-hand-sorting, both for total abundance and for the three ecological categories of earthworms. No significant effect was observed of the proportion of urban land cover, nor of the interaction with the sampling method, on total earthworm abundance or species richness. No interaction was observed of the soil parameters studied (soil organic matter, texture or water holding capacity) with the sampling method. It is concluded that, in urbanized environments, where due to social and health and safety considerations it is not always possible to use chemicals and to disturb land by excavating soil pits, the electrical method is a useful tool for sampling earthworms.

International audience; The SETA group (Agricultural Science and Technology) sent a letter to the Italian Parliament advising them not to adopt a legislative measure that favors organic and biodynamic agriculture compared to the conventional one. Another group of scientists thinks instead that the Italian government should favor organic and biodynamic agriculture and explains its reasons. The current article lists the SETA letter and the point of view of other Italian and international ecologists, economists, pedologists, ecologists, writers and artists, which are tendentially “protectors of nature” but not idealists. Nor are they so much in agreement with each other. Judge for yourself in scrolling through the Discussion chapter. The divergence leads to the “why” it is necessary to switch to organic farming not on the “need” to do so. There are also disappointments about the use of GMOs orthe costs of highly technological agriculture. The situation is such on planet Earth that it is necessary to involve the whole society to get out of it. In addition, politicians are also needed to structure the collective action that only if conceived by the whole society and not by individuals or by non-coordinated organizations can save our species. We are convinced that agriculture and soil are among the fundamental levers of this action.

The risk assessment of pesticides on soil fauna is an issue to protect agroecosystem sustainability. Enchytraeids are recognized as relevant soil bioindicators of chemical stress in agroecosystems. In laboratory, the reproduction test was found to be sensitive to reveal chemical impacts on enchytraeids. However, it does not allow to assess the impacts on ecological functions in which enchytraeids are involved. The objectives of this study were (i) to explore the feasibility of the bait-lamina test with enchytraeids under laboratory conditions and (ii) to compare its sensitivity with the Enchytraeid Reproduction Test. We exposed individuals of Enchytraeus albidus to two commercial formulations of fungicides widely used in Europe. The EC50 reproduction for the Swing® Gold (50 g L−1 epoxiconazole and 133 g L−1 dimoxystrobin) and the Cuprafor micro® (50% copper oxychloride) were respectively estimated at 1.66 ± 0.93 times the recommended dose and > 496 mg kg−1 of copper. However, no impact was found on the feeding activity of enchytraeids. The bait-lamina test thus appeared less sensitive than the Enchytraeid Reproduction Test to the tested fungicides. Despite that, this test which is achievable under laboratory conditions and allows assessing indirect effects of pesticides is quick, cheap, and easy to perform. It would deserve to be used to explore longer-exposure effects through the repeated addition of bait-lamina sticks.

The frequent, widespread use of pesticides in agriculture adversely affects biodiversity, human health, and water quality. In 2008, the French government adopted an environmental policy plan, “Ecophyto 2018”, to halve pesticide use within 10 years. Trends in synthetic pesticide sales and use in France were described, through three different indicators: the number of unit doses (NUD), the quantity of active ingredient (QAI), and the treatment frequency index (TFI). Changes in pesticide use on seven of the principal arable crops in France since the implementation of this policy plan were analyzed, together with the impact of changes in pesticide use on water quality. No evidence was found for a decrease in pesticide sales at national level between 2008 and 2013. In terms of the TFI values for individual crops, the only decrease in pesticide use observed since 2001 was for soft wheat. This decrease was very slight, and pesticide use did not decline more rapidly after 2006 than before. Changes in pesticide use differed between French regions and crops. Water pollution did not decrease during the period studied. Possible explanations for the lack of effectiveness of the French environmental plan are considered in the context of European legislation.

Earthworms act synergistically with microorganisms in soils. They are ecosystem engineers involved in soil organic matter degradation and nutrient cycling, leading to the modulation of resource availability for all soil organisms. Using a soil microcosm approach, we aimed to assess the influence of the earthworm Aporrectodea caliginosa on the response of soil microbial activities against two fungicides, i.e., Cuprafor Micro® (copper oxychloride, a metal) and Swing® Gold (epoxiconazole and dimoxystrobin, synthetic organic compounds). The potential nitrification activity (PNA) and soil enzyme activities (glucosidase, phosphatase, arylamidase, and urease) involved in biogeochemical cycling were measured at the end of the incubation period, together with earthworm biomass. Two common indices of the soil biochemistry were used to aggregate the response of the soil microbial functioning: the geometric mean (Gmean) and the Soil Quality Index (SQI). At the end of the experiment, the earthworm biomass was not impacted by the fungicide treatments. Overall, in the earthworm-free soil microcosms, the two fungicides significantly increased several soil enzyme and nitrification activities, leading to a higher GMean index as compared to the non-treated control soils. The microbial activity responses depended on the type of activity (nitrification was the most sensitive one), on the fungicide (Swing® Gold or Cuprafor Micro®), and on the doses. The SQI indices revealed higher effects of both fungicides on the soil microbial activity in the absence of earthworms. The presence of earthworms enhanced all soil microbial activities in both the control and fungicide-contaminated soils. Moreover, the magnitude of the fungicide impact, integrated through the SQI index, was mitigated by the presence of earthworms, conferring a higher stability of microbial functional diversity. Our results highlight the importance of biotic interactions in the response of indicators of soil functioning (i.e., microbial activity) to pesticides.

The use of pesticides in agroecosystems can have negative effects on earthworms, which play key roles in soil functioning such as organic matter decomposition. The aim of this study was to assess the effects of two fungicides (Cuprafor micro®, composed of copper oxychloride, and Swing Gold®, composed of epoxiconazole (EPX) and dimoxystrobin (DMX)) on earthworm reproduction by exposing adults and cocoons. First, adult Aporrectodea caliginosa individuals were exposed for 28 days to 3.33, 10 and 30 times the recommended dose (RD) of Cuprafor micro® corresponding to 25.8, 77.5 and 232.5 mg kg−1 dry soil of copper, respectively, and 0.33, 1 and 3 times the RD of Swing Gold® (corresponding to 5.2 × 10−2 mg DMX kg−1 + 1.94 × 10−2 mg EPX kg−1, 1.55 × 10−1 mg DMX kg−1 + 5.81 × 10−2 mg EPX kg−1 and 4.62 × 10−1 mg DMX kg−1 + 1.74 × 10−1 mg EPX kg−1 respectively), in addition to a control soil with no fungicide treatment. Cocoon variables (production, weight, hatching success, hatching time) were monitored. Second, “naive” cocoons produced by uncontaminated earthworms were exposed to soils contaminated by the same concentrations of the two fungicides, and we assessed hatching success and hatching time. In the first experiment, cocoon production was halved at the highest copper concentration (232.5 mg Cu kg−1 of dry soil) as compared to the control. Cocoons took 5 more days to hatch, and the hatching success decreased by 35% as compared to the control. In the Swing Gold® treatments, cocoon production was reduced by 63% at 3 times the RD, and the hatching success significantly decreased by 16% at the RD. In the second experiment, only the hatching success of cocoons was impacted by Swing Gold® at 3 times the RD (30% less hatching). It is concluded that the cocoon stock in the soil is crucial for the renewal of populations in the field. The most sensitive endpoint was the hatching success of the cocoons produced by exposed adults. This endpoint and the effects observed on the “naive” cocoons could be taken into account in pesticide risk assessment.

International audience; Belowground communities, and especially earthworms, provide numerous services in agroecosystems. In the current context of crop genetic erosion, this study aims at assessing the effect of wheat cultivar diversity on earthworm growth and survival. Our laboratory experiment was divided into two periods: (1) the “feeding period” (4 months), simulating conditions in autumn with cool temperatures and high food availability, and (2) the “non-feeding period” (1 month) simulating conditions in winter with cold temperatures and low food availability because litter had already been consumed without replacement. The aerial biomass of mixtures of wheat cultivars was used as food for earthworms and their growth was measured twice during the feeding period and once during the non-feeding period. We found two major effects on earthworm growth. Increasing the cultivar number (i) increased juvenile earthworm growth and earthworm survival and (ii) decreased adult earthworm growth. A consistent positive effect of nitrogen concentration of the biomass was found on growth and this effect was independent of biodiversity effects. This study suggests that the loss of litter intraspecific genetic diversity has an impact on earthworm populations, probably through mechanisms linked to the variance in edibility or the balanced diet hypotheses. While they remain to be tested in the field, these results suggest that the current losses of crop genetic diversity could potentially impact processes at the community or ecosystem level through a decrease in food quality.

International audience; Recent EFSA (European Food Safety Authority) reports highlighted that the ecological risk assessment of pesticides needed to go further by taking more into account the impacts of chemicals on biodiversity under field conditions. We assessed the effects of two commercial formulations of fungicides separately and in mixture, i.e., Cuprafor Micro® (containing 500 g kg−1 copper oxychloride) at 4 (C1, corresponding to 3.1 mg kg−1 dry soil of copper) and 40 kg ha−1 (C10), and Swing® Gold (50 g L−1 epoxiconazole EPX and 133 g L−1 dimoxystrobin DMX) at one (D1, 5.81 10−2 and 1.55 10−1 mg kg−1 dry soil of EPX and DMX, respectively) and ten times (D10) the recommended field rate, on earthworms at 1, 6, 12, 18 and 24 months after the application following the international ISO standard no. 11268-3 to determine the effects on earthworms in field situations. The D10 treatment significantly reduced the species diversity (Shannon diversity index, 54% of the control), anecic abundance (29% of the control), and total biomass (49% of the control) over the first 18 months of experiment. The Shannon diversity index also decreased in the mixture treatment (both fungicides at the recommended dose) at 1 and 6 months after the first application (68% of the control at both sampling dates), and in C10 (78% of the control) at 18 months compared with the control. Lumbricus terrestris, Aporrectodea caliginosa, Aporrectodea giardi, Aporrectodea longa, and Allolobophora chlorotica were (in decreasing order) the most sensitive species to the tested fungicides. This study not only addressed field ecotoxicological effects of fungicides at the community level and ecological recovery, but it also pinpointed some methodological weaknesses (e.g., regarding fungicide concentrations in soil and statistics) of the guideline to determine the effects on earthworms in field situations.

International audience; Long-term organic fertilizer (OF) application on agricultural soils is known to induce soil Cu and Zn contamination, along with pH and organic matter changes, which in turn alter the soil Cu and Zn availability. Our study was aimed at assessing Cu and Zn availability in long-term OF-amended soils by distinguishing the importance of increased contamination levels versus pH and organic matter changes in soil. Seventy-four soil samples were collected over time from fields corresponding to three soil types upon which no, mineral, or organic fertilization had been applied over a decade, and thus exhibited a gradient of Cu and Zn contamination, pH, and organic matter concentration. Soil Cu and Zn contamination (i.e. total and DTPA-extractable Cu and Zn concentration), soil solution chemistry (i.e. pH and dissolved organic matter concentration and aromaticity) and Cu and Zn availability (i.e. total concentration and free ionic activity in solution and DGT-available concentration in soil) levels were measured. The Windermere humic aqueous model (WHAM) was used to estimate Zn2+ activity and dissolved organic matter (DOM) binding properties in soil solution. Regardless of the soil type, organic fertilization increased Cu and Zn contamination in soil, in addition to the pH and the DOM concentration, aromaticity and binding properties in soil solution. The pH increase prompted a decrease in the total Zn concentration and Zn2+ activity in soil solution. The concomitant pH increase and DOM concentration, aromaticity and binding properties boosted the total Cu concentration but decreased the Cu2+ activity in soil solution. DGT-available Cu and Zn varied very little between the three fertilization modalities. Our results suggest that pH and DOM changes were able to regulate Cu and Zn availability in long-term OF amended soils by exerting a protective effect that offset the concomitant increase in soil Cu and Zn contamination.

Agricultural intensification, and in particular the use of pesticides, leads over the years to a loss of biodiversity and a decline of ecosystem services in cultivated zones and agricultural landscapes. Among the animal communities involved in the functioning of agro-ecosystems, earthworms are ubiquitous and recognized as indicators of land uses and cultural practices. However, little data is available on the levels of pesticides in such organisms in natura, which would allow estimating their actual exposure and the potentially resulting impacts. Thus, the objective of this study was to develop a sensitive analytical methodology to detect and quantify 27 currently used pesticides in earthworms (Allolobophora chlorotica). A modified QuEChERS extraction was implemented on individual earthworms. This step was followed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The whole analytical method was validated on spiked earthworm blank samples, with regard to linearity (from 1 to 100 method limit of quantification, r2 > 0.95), intra-day precision (relative standard deviation (RSD)

How pesticides are affecting earthworms? . 6th Symposium on sustainable use of pesticides Directive (SUD) - Integrated Pest Management, PAN Europe

Because of the wide use of pesticides in agriculture, there is still a need of higher-tier field studies to assess ecotoxicological effects of commercial formulations on a wider variety of non-target soil organisms such as soil annelids. We here tested the effects of different concentrations of two fungicide formulations, i.e., Cuprafor Micro(A (R)) (composed of 500 g kg(-1) copper oxychloride) and Swing Gold(A (R)) (composed of 50 g l(-1) epoxiconazole and 133 g l(-1) dimoxystrobin) on two families of terrestrial oligochaetes (Lumbricidae and Enchytraeidae) after 1 month of exposure. We also assessed the feeding activity of soil organisms using the bait lamina method. Along with the feeding activity, the enchytraeid density, diversity and communities were not different in the control and the contaminated plots. By contrast, epigeic earthworms were absent and earthworm diversity and densities of anecic species decreased significantly in the plots contaminated at ten times the recommended dose of the Swing Gold(A (R)) formulation. The copper fungicide (at 0.75 and 7.5 kg Cu ha(-1)) and the treatment with the pesticide mixture (Cuprafor Micro(A (R)) at 0.75 kg Cu ha(-1) and Swing Gold(A (R)) at the recommended dose) did not affect Oligochaeta communities compared with the control, except the Shannon index for earthworms in the mixture of both fungicides. Responses of the two annelid families to the tested pesticides were different with higher effects observed on the diversity and the community structure of earthworms compared with enchytraeids. This study allowed detecting early changes on oligochaete populations after pesticide application.

France can be qualified as terra incognita regarding terrestrial enchytraeids because very little data has been recorded so far in this country. In spring and autumn 2016, enchytraeid communities were investigated in a loamy soil in a meadow located in the park of Versailles palace, France. In total, twenty four enchytraeid species were identified, belonging to six different genera i.e. eleven Fridericia species, four Enchytraeus species, four Achaeta species, two Buchholzia species, two Marionina species and one Enchytronia species. According to the published data, this was one of the highest diversity found in a meadow in Europe.

Exotic peregrine earthworms are often considered to cause environmental harm and to have a negative impact on native species, but, as ecosystem engineers, they enhance soil physical properties. Pontoscolex corethrurus is by far the most studied morphospecies and is also the most widespread in tropical areas. The term of morphospecies is used in this review because P. corethrurus may in fact constitute a complex of cryptic species. This earthworm is found in a wide range of habitats, from apparently pristine to any kind of human-disturbed environment. This review synthesizes 265 studies describing the distribution, morphology, biological and ecological traits of this morphospecies, as well as its impacts on soil conditions and communities. We then discuss the characteristics necessary for this specific morphospecies to become a successful colonizer throughout the world and the positive and negative effects it can have on the ecosystems that it has invaded. We emphasize the lack of knowledge of P. corethrurus reproductive mode and ploidy level, of its population genetics, and of the potential existence of cryptic species. To finish, we highlight the fact that data on P. corethrurus interactions with non-earthworm soil macrofauna are scarce.

Ecotoxicological tests with earthworms are widely used and are mandatory for the risk assessment of pesticides prior to registration and commercial use. The current model species for standardized tests is Eisenia fetida or Eisenia andrei. However, these species are absent from agricultural soils and often less sensitive to pesticides than other earthworm species found in mineral soils. To move towards a better assessment of pesticide effects on non-target organisms, there is a need to perform a posteriori tests using relevant species. The endogeic species Aporrectodea caliginosa (Savigny, 1826) is representative of cultivated fields in temperate regions and is suggested as a relevant model test species. After providing information on its taxonomy, biology, and ecology, we reviewed current knowledge concerning its sensitivity towards pesticides. Moreover, we highlighted research gaps and promising perspectives. Finally, advice and recommendations are given for the establishment of laboratory cultures and experiments using this soil-dwelling earthworm species. Electronic supplementary material The online version of this article (10.1007/s11356-018-2579-9) contains supplementary material, which is available to authorized users.

Biochars are used as amendments to improve soil quality, but their effects on edaphic organisms such as earthworms remain controversial. This study aimed to assess the effects of adding a poultry manure-derived biochar into a contaminated technosol on trace element (TE) (i.e. As, Cd, Cu, Pb, and Zn) bioavailability for two earthworm species, Aporrectodea icterica and Aporrectodea longa. Three components of the bioavailability concept were determined using a pot experiment: (1) total soil TE (potentially reactive) and TE concentrations in the soil pore water (environmental availability), (2) TE concentrations in depurated whole earthworm bodies (environmental bioavailability) and (3) ecophysiological and biochemical effects on earthworms (toxicological bioavailability). Biochar addition increased TE concentrations in the soil pore water respectively from 1.8, 2.7, 9.4, 0.7 and 959 to 6, 6.2, 19.3, 6.9, and 3003 µg L−1 for As, Cd, Cu, Pb and Zn. Biochar addition did not influence TE environmental bioavailability for earthworms, except a decreased As concentration (32.5 to 15.2 µg g−1) in A. icterica. This suggests an inter-specific variability in As homeostasis in the Aporrectodea genus. In line with this internal As decrease, the Glutathione-S-transferase (GST) activity decreased by 42% and protein and lipid contents slightly increased (14 and 25%, respectively) in A. icterica tissues. The body weight of both earthworm species decreased for the biochar-amended soil. Environmental TE availability depended on both the biochar addition and the earthworm activity in the contaminated soil, while environmental and toxicological bioavailabilities resulted from the earthworm species, the targeted TE and biochar supply to the soil.

Développement d’une méthode multi-résidus par LC-MS/MS pour la quantification de traces de pesticides dans les vers de terre. 47ème congrès du Groupe Français des Pesticides

10 articles; Fate and impact of pesticides: new directions to explore

Enchytraeids, also called potworms, are Annelida Oligochaeta. They are present in many different soils worldwide and were found to be dominant in biomass or abundance in many habitats. These organisms are involved in key soil functions such as the dynamics of soil structure and organic matter. Based on an exhaustive literature review, we summarized the ecology and main roles of enchytraeids in soils. We also showed that enchytraeids are sensitive to changes in land use and management, both in terms of total abundance and community composition. The knowledge gaps were highlighted, as well as the needs for further research allowing to better understand the functional importance of these organisms in a context of global changes and anthropic pressures.

National audience; La compréhension des mécanismes d'effets toxiques et l'évaluation du risque des substances pour les écosystèmes sont des enjeux majeurs des recherches en écotoxicologie. Les effets écotoxicologiques : de la molécule à la population présente, au travers d'exemples concrets et de résultats de recherches récents, des concepts, des méthodes et des outils de mesure et de modélisation, qui permettent de comprendre, d'expliquer et de prédire l'impact des polluants chimiques de la cellule à la population. Cet ouvrage illustre la complexité et la richesse des questions scientifiques concernant les impacts toxiques sur différents niveaux d'organisation biologique. Il apporte des réponses aux lecteurs intéressés par les enjeux de compréhension et de prédiction des risques toxiques chimiques.

Tillage influences first soil structure and then soil organisms such as earthworms, which are highly involved in the creation of soil macroporosity. This study assessed the temporal dynamics of earthworm-related macroporosity in two ploughed and one unploughed cropping systems. Three sampling dates were chosen: one month before ploughing, and one and five months after the event. Earthworm communities, able to rebuild the macroporosity, were sampled one and five months after ploughing. Before ploughing, the burrow continuity, i.e., the number of burrows with a vertical length greater than 5 cm, was not significantly different in the three systems. It was stable between the three sampling dates in the unploughed system but it decreased by 65% and 46% after ploughing (i.e., in two months) in the organic and the conventional systems, respectively. Five months after ploughing, the burrow continuity remained between two and four times lower than in the unploughed living mulch cropping system. Earthworm biomass was higher and burrow system characteristics (i.e. burrow volume and continuity) were more stable over time in the non-tilled with a living mulch cropping system than in the tilled systems. Earthworm-produced macroporosity was thus substantially decreased after ploughing in conventional and organic systems and had still not totally recovered 5 months later. This can lead to large functional consequences on soil structure and thus on air and water fluxes in the soil.

The use of pesticides in crop fields may have negative effects on soil Oligochaeta Annelida, i.e., earthworms and enchytraeids, and thus affect soil quality. The aim of this study was to assess the effects of two commercial fungicide formulations on the earthworm Aporrectodea caliginosa and the enchytraeid Enchytraeus albidus in a natural soil. The fungicides were Cuprafor micro® (copper oxychloride), commonly used in organic farming, and Swing Gold® (epoxiconazole and dimoxystrobin), a synthetic fungicide widely used in conventional farming to protect cereal crops. Laboratory experiments were used to assess the survival, biomass loss and avoidance behaviour. No lethal effect was observed following exposure to the copper fungicide for 14 days, even at 5000 mg kg−1 of copper, i.e. 650 times the recommended dose (RD). However, a significant decrease in biomass was observed from 50 mg kg−1 of copper (6.5 times the RD) for A. caliginosa and at 5000 mg kg−1 of copper (650 times the RD) for E. albidus. These sublethal effects suggest that a longer period of exposure would probably have led to lethal effects. The EC50 avoidance for the copper fungicide was estimated to be 51.2 mg kg−1 of copper (6.7 times the RD) for A. caliginosa, and 393 mg kg−1 of copper (51 times the RD) for E. albidus. For the Swing Gold® fungicide, the estimated LC50 was 7.0 10−3 mL kg−1 (6.3 times the RD) for A. caliginosa and 12.7 10−3 mL kg−1 (11.0 times the RD) for E. albidus. No effect on biomass or avoidance was observed at sublethal concentrations of this synthetic fungicide. It was concluded that enchytraeids were less sensitive than earthworms to the two commercial fungicides in terms of mortality, biomass loss and avoidance behaviour. Therefore we discuss the different strategies possibly used by the two Oligochaeta species to cope with the presence of the pesticides were discussed, along with the potential consequences on the soil functions.

In soil, the determination of total concentration using an exhaustive extraction method is not relevant to evaluate the exposure of an organism to a chemical, because of sorption processes. However, little attention has been paid to the bioavailability of organic contaminants to earthworms, and to their effect on sensitive earthworm species found in cultivated fields, such as Aporrectodea icterica [1]. As ecosystem engineers, earthworms influence soil structure and chemistry and are thus key-organisms for the health of terrestrial ecosystems. However, they can be threatened by contaminants such as the fungicide epoxiconazole, which is persistent in soil and presents a chronic toxicity even towards a resistant earthworm species (Eisenia fetida) at a concentration close to the recommended dose. This study aimed to propose a mild extraction method to evaluate the bioavailability of the fungicide epoxiconazole to the earthworm Aporrectodea icterica [2]. Experiments were conducted in soils presenting various textures and organic carbon contents, spiked with formulated epoxiconazole 7 to 56 days prior to their extraction at one-fold to ten-fold the recommended dose. In parallel, the epoxiconazole concentration was determined in exposed earthworms. All the analysis were performed by UHPLCMS/ MS. The fungicide’s effects were evaluated by measuring weight gain, enzymatic activities and total protein contents. The proposed method to evaluate bioavailability was based on the pesticide entrapment in the cavity of hydroxypropyl-?-cyclodextrin, thus fulfilling the criteria of the ISO 17402 norm [3]. Furthermore, this mild method was demonstrated to be sensitive to soil sorption capacities and to ageing. The mild extraction method was then applied to explore the relationship between total and bioavailable concentrations in soil and in A. icterica, over 7 or 28 days exposure time. This demonstrated the proportionality between epoxiconazole concentration in earthworm and available in soil (up to 96%, with regression coefficient R2 0.98). Sublethal effects on earthworm remained not significant. References 1. C. Pelosi, S. Joimel, D. Makowski, 2013. Chemosphere 90, 895-900 2. S. Nélieu, G. Delarue, E. Ollivier, P. Awad, F. Fraillon, C. Pelosi, Environ. Sci. Pollut. Res. In Press DOI 10.1007/s11356-015-5270-4 3. ISO 17402, 2008. Soil quality - Guidance for the selection and application of methods for the assessment of bioavailability of contaminants in soil and soil materials

Background and aims Rapeseed ( Brassica napus L.) is a Cd/Zn-accumulator whereas soil conditioners such as biochars may immobilize trace elements. These potentially complementary soil remediation options were trialed, singly and in combination, in a pot experiment with a metal( loid )-contaminated technosol. Methods The technosol [total content in mg kg −1 Zn 6089, Cd 9.4, Cu 110, and Pb 956] was either amended (2% w/w) or not with a poultry manure-derived biochar. Rapeseed was cultivated for both soil treatments during 24 weeks up to harvest under controlled conditions. Results Biochar incorporation into the technosol promoted the As, Cd, Cu, Mo, Ni, Pb and Zn solubility. It decreased foliar B, Cu and Mo concentrations, and Mo concentration in stems, pericarps and seeds. But, it did not impact neither the biomass of aerial rapeseed parts (except a decrease for seeds), nor their C (except a decrease for stems), seed fatty acid, seed starch and soluble sugar contents, and antioxidant capacity in both leaves and seeds. Biochar amendment increased the phytoextraction by aerial plant parts for K, P, and S, reduced it for N, Ca, B, Mo, Ni and Se, whereas it remained steady for Mg, Zn, Fe, Mn, Cu, Cd and Co. Conclusions The biochar incorporation into this technosol did not promote Cd, Cu and Zn phytoextraction by rapeseed and its potential oilseed production, but increased the solubility of several metal( loid )s. Here Zn and Cd concentrations in the soil pore water were decreased by rapeseed, showing the feasibility to strip available soil Zn and Cd in combination with seed production.

International audience; No-till has been proposed to limit the negative impacts of intensive agriculture. Soil organisms such as earthworms are good indicators of soil tillage effects. The aim of this study was to assess the dynamics of both taxonomic and functional diversity of earthworm communities after switching from ploughing i.e. deep tillage at 25–30 cm depth inducing soil inversion in this paper, to no-till. We assumed (i) that this transition led to increased diversity and (ii) that both taxonomic and functional indices indicated changes in tillage practices. Earthworms were sampled at three experimental sites in France on a Cambisol, a Luvisol, and a Fluvisol over several years (from 1995 to 2011), comparing both tillage systems. Standardized mean differences (Cohen’s indices) were calculated on several taxonomic and functional indices to quantify the influence of tillage practices within each site. We generally found a neutral or positive effect of the no-till on taxonomic and functional indices. The temporal responses of earthworm diversity were stronger in earliest stages (less than 6 years) than in oldest ones (more than 8 years) revealing transitory effects on diversity. In site III, for instance, a positive effect of no-till was observed on species and functional richness during the first 5 years but a negative effect was shown in year 6. Thus, no clear continuous increase of taxonomic and functional diversity was found in no-tilled fields, but positive and negative effects according to sites and years. More marked differences were found for taxonomic than for functional indices, suggesting that individuals settling the fields shared a similar combination of functional traits to those previously present. As transitory effects have been observed, this study highlighted that long-term trials are needed to assess the effects of cropping systems on earthworm biodiversity

National audience