The use of copper as plant protection product contributes to environmental contamination and resulting impacts on terrestrial and aquatic biodiversity and ecosystem functions.

Copper-based plant protection products (PPPs) are widely used in both conventional and organic farming, and to a lesser extent for non-agricultural maintenance of gardens, greenspaces, and infrastructures. The use of copper PPPs adds to environmental contamination by this trace element. This paper aims to review the contribution of these PPPs to the contamination of soils and waters by copper in the context of France (which can be extrapolated to most of the European countries), and the resulting impacts on terrestrial and aquatic biodiversity, as well as on ecosystem functions. It was produced in the framework of a collective scientific assessment on the impacts of PPPs on biodiversity and ecosystem services in France. Current science shows that copper, which persists in soils, can partially transfer to adjacent aquatic environments (surface water and sediment) and ultimately to the marine environment. This widespread contamination impacts biodiversity and ecosystem functions, chiefly through its effects on phototrophic and heterotrophic microbial communities, and terrestrial and aquatic invertebrates. Its effects on other biological groups and biotic interactions remain relatively under-documented.

Impacts of neonicotinoids on biodiversity: a critical review.

Neonicotinoids are the most widely used class of insecticides in the world, but they have raised numerous concerns regarding their effects on biodiversity. Thus, the objective of this work was to do a critical review of the contamination of the environment (soil, water, air, biota) by neonicotinoids (acetamiprid, clothianidin, imidacloprid, thiacloprid, thiamethoxam) and of their impacts on terrestrial and aquatic biodiversity. Neonicotinoids are very frequently detected in soils and in freshwater, and they are also found in the air. They have only been recently monitored in coastal and marine environments, but some studies already reported the presence of imidacloprid and thiamethoxam in transitional or semi-enclosed ecosystems (lagoons, bays, and estuaries). The contamination of the environment leads to the exposure and to the contamination of non-target organisms and to negative effects on biodiversity. Direct impacts of neonicotinoids are mainly reported on terrestrial invertebrates (e.g., pollinators, natural enemies, earthworms) and vertebrates (e.g., birds) and on aquatic invertebrates (e.g., arthropods). Impacts on aquatic vertebrate populations and communities, as well as on microorganisms, are less documented. In addition to their toxicity to directly exposed organisms, neonicotinoid induce indirect effects via trophic cascades as demonstrated in several species (terrestrial and aquatic invertebrates). However, more data are needed to reach firmer conclusions and to get a clearer picture of such indirect effects. Finally, we identified specific knowledge gaps that need to be filled to better understand the effects of neonicotinoids on terrestrial, freshwater, and marine organisms, as well as on ecosystem services associated with these biotas.

Linking ecotoxicological effects on biodiversity and ecosystem functions to impairment of ecosystem services is a challenge: an illustration with the case of plant protection products.

There is growing interest in using the ecosystem services framework for environmental risk assessments of chemicals, including plant protection products (PPPs). Although this topic is increasingly discussed in the recent scientific literature, there is still a substantial gap between most ecotoxicological studies and a solid evaluation of potential ecotoxicological consequences on ecosystem services. This was recently highlighted by a collective scientific assessment (CSA) performed by 46 scientific experts who analyzed the international science on the impacts of PPPs on biodiversity, ecosystem functions, and ecosystem services. Here, we first point out the main obstacles to better linking knowledge on the ecotoxicological effects of PPPs on biodiversity and ecological processes with ecosystem functions and services. Then, we go on to propose and discuss possible pathways for related improvements. We describe the main processes governing the relationships between biodiversity, ecological processes, and ecosystem functions in response to effects of PPP, and we define categories of ecosystem functions that could be directly linked with the ecological processes used as functional endpoints in investigations on the ecotoxicology of PPPs. We then explore perceptions on the possible links between these categories of ecosystem functions and ecosystem services among a sub-panel of the scientific experts from various fields of environmental science. We find that these direct and indirect linkages still need clarification. This paper, which reflects the difficulties faced by the multidisciplinary group of researchers involved in the CSA, suggests that the current gap between most ecotoxicological studies and a solid potential evaluation of ecotoxicological consequences on ecosystem services could be partially addressed if concepts and definitions related to ecological processes, ecosystem functions, and ecosystem services were more widely accepted and shared within the ecotoxicology community. Narrowing this gap would help harmonize and extend the science that informs decision-making and policy-making, and ultimately help to better address the trade-off between social benefits and environmental losses caused by the use of PPPs.

Main conclusions and perspectives from the collective scientific assessment of the effects of plant protection products on biodiversity and ecosystem services along the land-sea continuum in France and French overseas territories.

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.

Changes in soil carbon mineralization related to earthworm activity depend on the time since inoculation and their density in soil.

Earthworms play a key role in soil carbon mineralization, but their effect is highly uncertain and suspected to vary as a function of several factors, particularly the earthworm density and time from earthworm inoculation. We conducted a meta-analysis considering these factors based on 42 experiments comparing carbon mineralization in the absence and presence of earthworms at different times. The results reveal an average carbon mineralization increase of 24% (sd 41%) in the presence of earthworms with an initial median earthworm density of 1.95 mg/g soil DM (Dry Mass) (sd 48%). We show that carbon mineralization due to earthworms was related to their density and time from inoculation. From a simple regression model using these two variables, the estimated impact of earthworms on carbon mineralization was 20% increase from 0 to 60 days and 14% decrease at day 350 for a density of worms commonly found in soils (0.5 mg/g soil DM). Finally, we proposed a simple equation that could be used in organic matter decomposition models that do not take macrofauna into account.

Analysis of complex trophic networks reveals the signature of land-use intensification on soil communities in agroecosystems.

Increasing evidence suggests that agricultural intensification is a threat to many groups of soil biota, but how the impacts of land-use intensity on soil organisms translate into changes in comprehensive soil interaction networks remains unclear. Here for the first time, we use environmental DNA to examine total soil multi-trophic diversity and food web structure for temperate agroecosystems along a gradient of land-use intensity. We tested for response patterns in key properties of the soil food webs in sixteen fields ranging from arable crops to grazed permanent grasslands as part of a long-term management experiment. We found that agricultural intensification drives reductions in trophic group diversity, although taxa richness remained unchanged. Intensification generally reduced the complexity and connectance of soil interaction networks and induced consistent changes in energy pathways, but the magnitude of management-induced changes depended on the variable considered. Average path length (an indicator of food web redundancy and resilience) did not respond to our management intensity gradient. Moreover, turnover of network structure showed little response to increasing management intensity. Our data demonstrates the importance of considering different facets of trophic networks for a clearer understanding of agriculture-biodiversity relationships, with implications for nature-based solutions and sustainable agriculture.

Agroecosystem diversification with legumes or non-legumes improves differently soil fertility according to soil type.

Plant diversification through crop rotation or agroforestry is a promising way to improve sustainability of agroecosystems. Nonetheless, criteria to select the most suitable plant communities for agroecosystems diversification facing contrasting environmental constraints need to be refined. Here, we compared the impacts of 24 different plant communities on soil fertility across six tropical agroecosystems: either on highly weathered Ferralsols, with strong P limitation, or on partially weathered soils derived from volcanic material, with major N limitation. In each agroecosystem, we tested several plant communities for diversification, as compared to a matching low diversity management for their cropping system. Plant residue restitution, N, P and lignin contents were measured for each plant community. In parallel, the soil under each community was analyzed for organic C and N, inorganic N, Olsen P, soil pH and nematode community composition. Soil potential fertility was assessed with plant bioassays under greenhouse controlled climatic conditions. Overall, plant diversification had a positive effect on soil fertility across all sites, with contrasting effects depending on soil type and legumes presence in the community. Communities with legumes improved soil fertility indicators of volcanic soils, which was demonstrated through significantly higher plant biomass production in the bioassays (+18%) and soil inorganic N (+26%) compared to the low diversity management. Contrastingly, communities without legumes were the most beneficial in Ferralsols, with increases in plant biomass production in the bioassays (+39%), soil Olsen P (+46%), soil C (+26%), and pH (+5%). Piecewise structural equation models with Shipley's test revealed that plant diversification impacts on volcanic soil fertility were related to soil N availability, driven by litter N. Meanwhile, Ferralsols fertility was related to soil P availability, driven by litter P. These findings underline the importance of multifactorial and multi-sites experiments to inform trait-based frameworks used in designing optimal plant diversification in agroecological systems.

Cascading effects of moth outbreaks on subarctic soil food webs.

The increasing severity and frequency of natural disturbances requires a better understanding of their effects on all compartments of biodiversity. In Northern Fennoscandia, recent large-scale moth outbreaks have led to an abrupt change in plant communities from birch forests dominated by dwarf shrubs to grass-dominated systems. However, the indirect effects on the belowground compartment remained unclear. Here, we combined eDNA surveys of multiple trophic groups with network analyses to demonstrate that moth defoliation has far-reaching consequences on soil food webs. Following this disturbance, diversity and relative abundance of certain trophic groups declined (e.g., ectomycorrhizal fungi), while many others expanded (e.g., bacterivores and omnivores) making soil food webs more diverse and structurally different. Overall, the direct and indirect consequences of moth outbreaks increased belowground diversity at different trophic levels. Our results highlight that a holistic view of ecosystems improves our understanding of cascading effects of major disturbances on soil food webs.

Relationships between metal compartmentalization and biomarkers in earthworms exposed to field-contaminated soils.

Partitioning tissue metal concentration into subcellular compartments reflecting toxicologically available pools may provide good descriptors of the toxicological effects of metals on organisms. Here we investigated the relationships between internal compartmentalization of Cd, Pb and Zn and biomarker responses in a model soil organism: the earthworm. The aim of this study was to identify metal fractions reflecting the toxic pressure in an endogeic, naturally occurring earthworm species (Aporrectodea caliginosa) exposed to realistic field-contaminated soils. After a 21 days exposure experiment to 31 field-contaminated soils, Cd, Pb and Zn concentrations in earthworms and in three subcellular fractions (cytosol, debris and granules) were quantified. Different biomarkers were measured: the expression of a metallothionein gene (mt), the activity of catalase (CAT) and of glutathione-s-transferase (GST), and the protein, lipid and glycogen reserves. Biomarkers were further combined into an integrated biomarker index (IBR). The subcellular fractionation provided better predictors of biomarkers than the total internal contents hence supporting its use when assessing toxicological bioavailability of metals to earthworms. The most soluble internal pools of metals were not always the best predictors of biomarker responses. metallothionein expression responded to increasing concentrations of Cd in the insoluble fraction (debris + granules). Protein and glycogen contents were also mainly related to Cd and Pb in the insoluble fraction. On the other hand, GST activity was better explained by Pb in the cytosolic fraction. CAT activity and lipid contents variations were not related to metal subcellular distribution. The IBR was best explained by both soluble and insoluble fractions of Pb and Cd. This study further extends the scope of mt expression as a robust and specific biomarker in an ecologically representative earthworm species exposed to field-contaminated soils. The genetic lineage of the individuals, assessed by DNA barcoding with cytochrome c oxidase subunit I, did not influence mt expression.

Urban and industrial land uses have a higher soil biological quality than expected from physicochemical quality.

Despite their importance both in soil functioning and as soil indicators, the response of microarthropods to various land uses is still unclear. The aim of this study is to assess the effect of land use on microarthropod diversity and determine whether a soil's biological quality follows the same physicochemical quality-based gradient from forest, agriculture-grassland, agriculture-arable land, vineyards, urban vegetable gardens to urban, industrial, traffic, mining and military areas. A database compiling the characteristics of 758 communities has been established. We calculated Collembola community indices including: species richness, Pielou's evenness index, collembolan life forms, the abundance of Collembola and of Acari, the Acari/Collembola abundance ratio, and the Collembolan ecomorphological index. Results show that agricultural land use was the most harmful for soil microarthropod biodiversity, whilst urban and industrial land uses give the same level of soil biological quality as forests do. Furthermore, differences between the proportions of Acari and ecomorphological groups were observed between land uses. This study, defining soil microarthropod diversity baselines for current land uses, should therefore help in managing and preserving soil microarthropod biodiversity, especially by supporting the preservation of soil quality.

A structural equation model of soil metal bioavailability to earthworms: confronting causal theory and observations using a laboratory exposure to field-contaminated soils.

Structural equation models (SEM) are increasingly used in ecology as multivariate analysis that can represent theoretical variables and address complex sets of hypotheses. Here we demonstrate the interest of SEM in ecotoxicology, more precisely to test the three-step concept of metal bioavailability to earthworms. The SEM modeled the three-step causal chain between environmental availability, environmental bioavailability and toxicological bioavailability. In the model, each step is an unmeasured (latent) variable reflected by several observed variables. In an exposure experiment designed specifically to test this SEM for Cd, Pb and Zn, Aporrectodea caliginosa was exposed to 31 agricultural field-contaminated soils. Chemical and biological measurements used included CaC12-extractable metal concentrations in soils, free ion concentration in soil solution as predicted by a geochemical model, dissolved metal concentration as predicted by a semi-mechanistic model, internal metal concentrations in total earthworms and in subcellular fractions, and several biomarkers. The observations verified the causal definition of Cd and Pb bioavailability in the SEM, but not for Zn. Several indicators consistently reflected the hypothetical causal definition and could thus be pertinent measurements of Cd and Pb bioavailability to earthworm in field-contaminated soils. SEM highlights that the metals present in the soil solution and easily extractable are not the main source of available metals for earthworms. This study further highlights SEM as a powerful tool that can handle natural ecosystem complexity, thus participating to the paradigm change in ecotoxicology from a bottom-up to a top-down approach.

PAH dynamics in roadside environments: Influence on the consistency of diagnostic ratio values and ecosystem contamination assessments.

Roadside contamination (of air, soils and organisms) by polycyclic aromatic hydrocarbons (PAHs) was examined in an arable field and a mature forest (central France). Measured contents accounted for minute fractions of the cumulative vehicular exhaust emissions. The fate of vehicular PAHs was affected by many factors, including: atmospheric load dispersal, deposition on soils and vegetation, incorporation into water and organic matter cycles, and accumulation in species. Given these empirical results, we evaluated the consistency of a set of well-known diagnostic ratios. This effort has revealed that: i) most diagnostic ratio values vary considerably across roadside samples, including exhaust emissions; and ii) the first few meters from the carriageway or the road verge/forest interface or remote areas where surface water accumulates actually define turning or inflection points in the ratio profiles. These variations constitute a major obstacle to delimitating the extent of roadside contamination due to PAHs, in addition to raising questions over the applicability of ratios routinely used to designate sources. New ratios, namely (Flt+BkF)/(Pyr+BbF) and (Flt+BkF+BghiP)/Σ10PAH, have been specifically developed to address this challenge. The higher consistencies exhibited among environmental compartments as well as between surface soil and exhaust emissions still yield differentiated values relative to several industrial sources.

Subcellular partitioning of metals in Aporrectodea caliginosa along a gradient of metal exposure in 31 field-contaminated soils.

Subcellular fractionation of metals in organisms was proposed as a better way to characterize metal bioaccumulation. Here we report the impact of a laboratory exposure to a wide range of field-metal contaminated soils on the subcellular partitioning of metals in the earthworm Aporrectodea caliginosa. Soils moderately contaminated were chosen to create a gradient of soil metal availability; covering ranges of both soil metal contents and of several soil parameters. Following exposure, Cd, Pb and Zn concentrations were determined both in total earthworm body and in three subcellular compartments: cytosolic, granular and debris fractions. Three distinct proxies of soil metal availability were investigated: CaCl2-extractable content dissolved content predicted by a semi-mechanistic model and free ion concentration predicted by a geochemical speciation model. Subcellular partitionings of Cd and Pb were modified along the gradient of metal exposure, while stable Zn partitioning reflected regulation processes. Cd subcellular distribution responded more strongly to increasing soil Cd concentration than the total internal content, when Pb subcellular distribution and total internal content were similarly affected. Free ion concentrations were better descriptors of Cd and Pb subcellular distribution than CaCl2 extractable and dissolved metal concentrations. However, free ion concentrations and soil total metal contents were equivalent descriptors of the subcellular partitioning of Cd and Pb because they were highly correlated. Considering lowly contaminated soils, our results raise the question of the added value of three proxies of metal availability compared to soil total metal content in the assessment of metal bioavailability to earthworm.

The impact of management strategies in apple orchards on the structural and functional diversity of epigeal spiders.

Apple orchards are agro-ecosystems managed with high levels of inputs and especially pesticides. Epigeal spider communities were sampled in three seasons using pitfall traps in 19 apple orchards with four different management strategies (abandoned, under organic, Integrated Pest Management or conventional protection) and thus significantly different pesticide usage. The abundance and diversity of the spider communities was the highest in abandoned orchards. Higher diversity and evenness values were the only difference in spider communities from the organic orchards compared to the other commercial orchards. The analysis of five ecological traits (proportion of aeronauts, type of diet, overwintering stages, body size and maternal care), however, clearly showed differences in the spiders from the organic orchards. The spider species in the other commercial orchards were smaller and have higher dispersal abilities. Seven bioindicator species were identified in abandoned orchards, two species in organic ones (only Lycosidae) and one species in conventional orchards (Linyphiidae).

A thesaurus for soil invertebrate trait-based approaches.

Soil invertebrates are known to be much involved in soil behaviour and therefore in the provision of ecosystem services. Functional trait-based approaches are methodologies which can be used to understand soil invertebrates' responses to their environment. They (i) improve the predictions and (ii) are less dependent on space and time. The way traits have been used recently has led to misunderstandings in the integration and interpretation of data. Trait semantics are especially concerned. The aim of this paper is to propose a thesaurus for soil invertebrate trait-based approaches. T-SITA, an Internet platform, is the first initiative to deal with the semantics of traits and ecological preferences for soil invertebrates. It reflects the agreement of a scientific expert community to fix semantic properties (e.g. definition) of approximately 100 traits and ecological preferences. In addition, T-SITA has been successfully linked with a fully operational database of soil invertebrate traits. Such a link enhances data integration and improves the scientific integrity of data.

Is there a relationship between earthworm energy reserves and metal availability after exposure to field-contaminated soils?

Generic biomarkers are needed to assess environmental risks in metal polluted soils. We assessed the strength of the relationship between earthworm energy reserves and metal availability under conditions of cocktail of metals at low doses and large range of soil parameters. Aporrectodea caliginosa was exposed in laboratory to a panel of soils differing in Cd, Pb and Zn total and available (CaCl2 and EDTA-extractable) concentrations, and in soil texture, pH, CEC and organic-C. Glycogen, protein and lipid contents were recorded in exposed worms. Glycogen contents were not linked to the explaining variables considered. Variable selection identified CaCl2 extractable metals concentrations and soil texture as the main factors affecting protein and lipid contents. The results showed opposite effects of Pb and Zn, high inter-individual variability of biomarkers and weak relationships with easily extractable metals. Our results support the lack of genericity of energy reserves in earthworms exposed to field-contaminated soils.

Functional traits of soil invertebrates as indicators for exposure to soil disturbance.

We tested a trait-based approach to link a soil disturbance to changes in invertebrate communities. Soils and macro-invertebrates were sampled in sandy soils contaminated by long-term wastewater irrigation, adding notably organic matter and trace metals (TM). We hypothesized that functional traits of invertebrates depict ways of exposure and that exposure routes relate to specific TM pools. Geophages and soft-body invertebrates were chosen to inform on exposure by ingestion or contact, respectively. Trait-based indices depicted more accurately effects of pollution than community density and diversity did. Exposure by ingestion had more deleterious effects than by contact. Both types of exposed invertebrates were influenced by TM, but geophages mainly responded to changes in soil organic matter contents. The trait-based approach requires to be applied in various conditions to uncorrelate specific TM impacts from those of other environmental factors.

ISTA 14--impact of antibiotics from pig slurry on soil microbial communities, including the basidiomycete Trametes versicolor.

Livestock slurry containing antibiotics is a source of contamination of agricultural soils, with possible effects on soil function and micro-organisms. Extracellular oxido-reductases and hydrolases from the fungus T. versicolor and fungal growth were monitored in liquid cultures in the presence of tetracycline, lincomycine, sulfadiazine and ciprofloxacin for 10 days, in order to assess the suitability of these enzymes as biomarkers. Among the conditions of treatment, statistical analysis demonstrated an increase in manganese-dependent peroxidase after exposure to sulfadiazine at 1 mg/L when compared with the control. Acid phosphatase activity was decreased by lincomycine at 1 or 10 mg/L. Conversely, ß-glucosidase activity increased in the presence of this antibiotic at 10 mg/L. In Terrestrial Model Ecosystems spiked with contaminated pig slurry, lincomycine at the concentration of 8 or 80 µg/kg dry soil, and ciprofloxacin at 250 ng/kg dry soil decreased the activity of soil dehydrogenase, when compared with a green slurry treatment, over 28-day incubations. Laccase activity was similarly decreased in the presence of the highest concentration of antibiotics. We determined bacterial and fungal biomasses using Q-PCR. Bacterial biomass was increased in the presence of lincomycine at 80 µg/kg whatever the time of exposure, and to a lesser extent in the presence of ciprofloxacin at 250 ng/kg, but only at day 28. In contrast, both antibiotics, whatever their concentrations, did not modify fungal biomass in soil. In conclusion, we were unable to demonstrate important effects of antibiotics at concentrations found in the agricultural environment.

Earthworm cast production as a new behavioural biomarker for toxicity testing.

There is currently a lack of ecotoxicity tests adapted to earthworm species of higher ecological relevance and whose endpoints could be directly related to their ecological role in the soil. We propose a new and relatively simple ecotoxicity test based on the estimation of cast production (CP) by Lumbricus terrestris under laboratory conditions. CP was found to be linearly correlated to earthworm biomass and to be greatly influenced by soil water content. Azinphos-methyl had no effect on CP at all the concentrations tested. Significant decreases were observed at the normal application rate for other pesticides with (imidacloprid, carbaryl, methomyl) or without (ethyl-parathion and chlorpyrifos-ethyl) a clear concentration-effect response. For the highest concentration tested, reduction in CP varied between 35 and 67%. CP is straightforward and rapidly measured and ecologically meaningful. We thus believe it to be of great use as an endpoint in ecotoxicity testing.

OprF polymorphism as a marker of ecological niche in Pseudomonas.

OprF is the major outer-membrane protein of Pseudomonas sensu stricto (rRNA group I). In addition to playing a role as porin, membrane structural protein and root adhesion, this pleiotropic protein shows a length polymorphism corresponding to two types of OprF, termed OprF type 1 and OprF type 2. In a previous work, all the P. fluorescens isolated from bulk soil (non-rhizospheric) were shown to possess oprF type 1, while all the clinical P. fluorescens isolates and most rhizospheric strains corresponded to type 2. In this study, we further investigated the relation between the OprF polymorphism and the ecological niche by developing a culture-independent approach (a ratio polymerase chain reaction) to measure the percentage of each oprF type in environmental DNA samples, including two different soils and three different cultured plants (flax, wheat and grassland). Although the proportions of oprF type 2 between rhizospheric samples were quite variable, they were always very significantly higher (P<0.001) than the proportions of oprF type 2 of the adjacent bulk soil where the vast majority of oprF (>95%) corresponded to type 1. We discuss the potential applications of this ecological fingerprint in an agronomic and taxonomic point of view.