The upcoming European Soil Monitoring Law: An effective instrument for the protection of terrestrial ecosystems?

Soils are a precious resource consistently placed under several threats and urgently in need of protection within a regulatory framework at the European level. Soils are central to the provision of environmental services as well as human existence on earth. The need to protect soil has been identified by several recent European strategies and fortunately, a specific European regulation for soil protection is on the way-the European Soil Monitoring Law (formerly: Soil Health Law). However, efforts need to ensure that the upcoming Soil Monitoring Law closes gaps between existing regulations for chemicals and acknowledges current European strategies for environmental protection and sustainability. This brief communication started from a fruitful discussion among SETAC Global Soils Interest Group members on a recent public consultation on the newly proposed Soil Monitoring Law of the European Commission and highlights critical points focusing on the chemical pollution of soils. We emphasize urgent needs such as the essential definition of a "healthy state" of soils; the implementation of a suitable set of indicators and quality standards for the description of physical, chemical, and biological states of soils; the enforcement of the "polluter-pays" principle; and the establishment of a Europe-wide monitoring program. Results from monitoring need to be fed back into regulatory frameworks, including the regulation of chemicals. Guidance documents for the risk assessment of chemicals are outdated and need to be updated. Finally, actions need to be taken to foster healthy soils, stop biodiversity decline, and ensure the functioning of ecosystem services for future generations. Integr Environ Assess Manag 2024;20:316-321. © 2023 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Incorporation of sediment- and soil-specific aspects in the Criteria for Reporting and Evaluating Ecotoxicity Data (CRED).

In environmental risk assessment either for registration purposes or for retrospective assessments of monitoring data, the hazard assessment is predominantly based on effect data from ecotoxicity studies. Most regulatory frameworks require studies used for risk assessment to be evaluated for reliability and relevance. Historically, the Klimisch methodology was used in many regulatory procedures where reliability needed to be evaluated. More recently, the Criteria for Reporting and Evaluating Ecotoxicity Data (CRED) have been developed for aquatic ecotoxicity studies, providing more detailed guidance on the evaluation and reporting of not only the reliability but also the relevance of a scientific study. Here, we discuss the application of the CRED methodology for assessing sediment and soil ecotoxicity studies, addressing important sediment- and soil-specific criteria that should be included as part of the CRED evaluation system. We also provide detailed recommendations for the design and reporting of sediment and soil toxicity studies that can be used by scientists and researchers wishing to contribute ecotoxicological data for effect assessments carried out within regulatory frameworks. Integr Environ Assess Manag 2024;00:1-13. © 2024 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

The Recolonization Concentration Concept: Using Avoidance Assays with Soil Organisms to Predict the Recolonization Potential of Contaminated Sites.

In this study the recolonization concentration concept for soil organisms is presented and validated. This concept is based on the empirically deduced avoidance-recolonization hypothesis, which shows a negative correlation between avoidance (ACx) and recolonization (RCx) (ACx = RC100-x) responses. The concept was validated in a two-step approach composed by (i) individual placement tests, to demonstrate the non-influence of individual placement in a dual chamber avoidance test and (ii) small scale gradient tests to demonstrate that the number of colonizers reaching a soil patch with a certain concentration is independent on their previous exposure to lower concentrations. Overall, data show that avoidance data can be used, when framed under the recolonization concentration concept, to evaluate the recolonization potential of contaminated sites. The recolonization concept is an important theoretical concept that when coupled with spatial modelling tools could be used to tackle the spatial and temporal recovery dynamics of contaminated soil.

Response addition is more protective of biogeochemical cycles of carbon and phosphorus compared to concentration addition.

In soils, enzymes are crucial to catalyzing reactions and cycling elements such as carbon (C), nitrogen (N), and phosphorus (P). Although these soil enzymes are sensitive to metals, they are often disregarded in risk assessments, and regulatory laws governing their existence are unclear. Nevertheless, there is a need to develop regulatory standards for metal mixtures that protect biogeochemical cycles because soil serve as a sink for metals and exposures occur as mixtures. Using a fixed ratio ray design, we investigated the effects of 5 single metals and 10 quinary mixtures of Zn, Cu, Ni, Pb, and Co metal oxides on two soil enzymes (i.e., acid phosphatases [ACP] and beta glucosidases [BGD]) in two acidic Canadian soils (S1: acid sandy forest soil, and S2: acid sandy arable soil), closely matched to EU REACH standard soils. Compared to BGD, ACP was generally the more sensitive enzyme to both the single metals and the metal mixtures. The effective concentration inhibiting 50% enzyme activity (EC50) estimates for single Cu (2.1-160.7 mmol kg-1) and Ni (12-272 mmol kg-1) showed that those were the most toxic to both enzymes in both soils. For metal mixtures, response addition (RA) was more conservative in predicting metal effects compared to concentration addition (CA). For both additivity models, antagonism was observed except at lower concentrations (≤10,000 mg/kg) where synergism was observed. At higher concentrations (>10,000 mg/kg), free and CaCl2 extractable Cu protected both enzymes against the toxicity of other metals in the mixture. The results suggest that assuming CA at concentrations less than EC50 does not protect biogeochemical cycling of C and P. And Cu in soil may protect soil enzymes from other toxic metals and thus may have an overall positive role.

Community effect concentrations as a new concept to easily incorporate community data in environmental effect assessment of complex metal mixtures.

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

Are structural and functional endpoints of soil communities similarly affected by metal mixtures? - A terrestrial model ecosystem approach.

Soils are habitat to a variety of flora and fauna in a linked ecosystem which provides essential ecosystem services. In soil, metals can accumulate at high concentrations, because of anthropogenic activities, leading to toxic effects, threatening the ecosystem and the services it provides. In most real-world contamination scenarios, metals occur as complex mixtures which can interact and produce different toxicity than predicted from individual metal data. Current regulatory guidelines are based on single species responses to individual metals and ignore indirect effects inherent to the inter-linked nature of ecosystems. Also, the evaluation of anthropogenic impacts to the soil communities is usually measured through structural endpoints (e.g. abundance) disregarding functional measurements (e.g. organic matter decomposition rates), which are often seen as tightly related, and thus, similarly affected. In this study we tested three mixture ratios of five metal oxides (lead, copper, nickel, zinc, cobalt) at three dose levels (Low, Med, High) in a terrestrial model ecosystem experiment and measured structural and functional endpoints. Exposure to metal mixtures for 16 weeks did not affect the microarthropod community, but produced severe effects on soil microbial activity (PNR and DHA) reducing activity below 50% compared to control levels, in all dosed treatments. Metal contamination also significantly affected feeding activity and organic matter decomposition, but effects were not as pronounced as on microbial activity. Data suggest that, in the risk assessment of metals and their mixtures, effects on ecosystem structure and functions must be considered to provide adequate environmental protection.

Metal oxides and annealed metals as alternatives to metal salts for fixed-ratio metal mixture ecotoxicity tests in soil.

In soil metal ecotoxicology research, dosing is usually performed with metal salts, followed by leaching to remove excess salinity. This process also removes some metals, affecting metal mixture ratios as different metals are removed by leaching at different rates. Consequently, alternative dosing methods must be considered for fixed ratio metal mixture research. In this study three different metal mixture dosing methods (nitrate, oxide and annealed metal dosing) were examined for metal concentrations and toxicity. In the nitrate metal dosing method leaching reduced total metal retention and was affected by soil pH and cation exchange capacity (CEC). Acidic soils 3.22 (pH 3.4, CEC 8 meq/100g) and WTRS (pH 4.6, CEC 16 meq/100g) lost more than 75 and 64% of their total metals to leaching respectively while Elora (6.7 pH, CEC 21 meq/100g) and KUBC (pH 5.6, CEC 28 meq/100g) with higher pH and CEC only lost 13.6% and 12.2% total metals respectively. Metal losses were highest for Ni, Zn and Co (46.0%, 63.7% and 48.4% metal loss respectively) whereas Pb and Cu (5.6% and 20.0% metal loss respectively) were mostly retained, affecting mixture ratios. Comparatively, oxide and annealed metal dosing which do not require leaching had higher total metal concentrations, closer to nominal doses and maintained better mixture ratios (percent of nominal concentrations for the oxide metal dosing were Pb = 109.9%, Cu = 84.6%, Ni = 101.9%, Zn = 82.3% and Co = 97.8% and for the annealed metal dosing were Pb = 81.7%, Cu = 80.3%, Ni = 100.5%, Zn = 89.2% and Co = 101.3%). Relative to their total metal concentrations, nitrate metal dosing (lowest metal concentrations) was the most toxic followed by metal oxides dosing while the annealed dosing method was generally non-toxic. Due to the lack of toxicity of the annealed metals and their higher dosing effort, metal oxides, are the most appropriate of the tested dosing methods, for fixed-ratio metal mixtures studies with soil invertebrates.

Single metal and metal mixture toxicity of five metals to Oppia nitens in five different Canadian soils.

Metal mixture toxicity across soil types is a daunting challenge to risk assessment. Here, we evaluated metal mixture toxicity in Oppia nitens, using ten fixed metal mixture ratios in five Canadian soils that closely matched some of the EU PNEC reference soils. Soils were dosed with five metals (Cu, Zn, Pb, Co, Ni) as single metals (ten concentrations) and as mixtures (eight concentrations). Synchronized adult mites were exposed to metals, with survival and reproduction assessed after 28 days. We found out that (i) the differences among soils in mite sensitivity and single metals were not consistent when mites were exposed to metal mixtures, (ii) assuming concentration addition, the mixture interaction factor (MIF) showed that single metal low effect levels excessively underestimated low level metal mixture effects (iii) Zn emerged as a protective metal in most mixtures, and (iv) Soil properties such as CEC, independent of effects on metal speciation, explained more of the variation than measured metals. This study suggests that metal risk assessment should be done on a case by case basis. Further work is needed to ensure that by protecting soil-dwelling organisms from single metals, the risk from metal mixtures is appropriately protected for.

The effects of complex metal oxide mixtures on three soil invertebrates with contrasting biological traits.

For regulatory purposes, the concentration addition model is the default first tier for assessing joint-action toxicity of metal mixtures. Although many researchers have evaluated binary and ternary mixtures, fewer have investigated joint-action toxicity in more complex mixtures, where deviations from additivity are more likely due to the greater number of potential interactions. In this study, we tested fixed ratios of five metals (lead, copper, nickel, zinc, cobalt) as metal oxide mixtures on three soil invertebrate species (Enchytraeus crypticus, Folsomia candida, Oppia nitens) at different dose effect levels (EC10-EC90) in an acid sandy forest and a loamy soil. Total metal concentrations for mixture ratios in soil did not explain or correlate with species responses. For F. candida, toxicity was linked to metal solubility, while for O. nitens and E. crypticus, toxicity did not correlate with total or extractable metals. In O. nitens and E. crypticus, however, soil ingestion could be an important route of exposure. Analysing the joint effect of metal mixtures, F. candida response was globally additive, while E. crypticus and O. nitens both presented synergistic effects at low-dose effect levels. Estimations at the EC50 level underestimated the deviations from additivity which were larger at higher and especially lower effect levels. Testing across different effect concentrations (EC10-EC90) was an important tool allowing the identification of these larger deviations from additivity outside the EC50 threshold. Considering most protection thresholds are set below the EC50 level, and it was in this low effect range where the highest synergisms were observed, risk assessment schemes should test additivity at the target protection level using representative test organisms.

Toxicity assessment of metal mixtures to soil enzymes is influenced by metal dosing method.

Metals are present as mixtures in the environment, yet testing such complex mixture poses design and technical challenges. One possible solution is the use of fixed ratios, i.e. rays of increasing metal concentrations. But fixed ratios rays are compromised when soils dosed with metal salts are leached due to metal-soil selectivity rules. Two alternative metal forms, metal oxides and spinel minerals of quinary metal mixtures (Pb, Cu, Co, Ni, Zn), were evaluated for their toxicity to soil microorganisms measured by the activity of ammonia monooxygenases and acid-phosphatases in three soils. Leaching, a required step for salts, had a larger effect on ammonia monooxygenases than metals. Generally, metal salts were the most toxic form, while the spinel minerals were the least toxic form. Two extractants, CaCl2 and DTPA, were evaluated for their ability to link toxicity to metals across all three metal forms. Salt toxicity was closely linked to CaCl2 extractable concentrations but DTPA was the most appropriate for oxides. We strongly recommend combining fixed ratio rays with metal oxides for metal mixture studies, since soil ratios created using oxides were more precise and required less experimental effort compared to salts and spinel minerals. Furthermore, because DTPA and CaCl2 closely tracked the toxicity of more realistic metal forms (i.e. oxides), we recommend that field studies investigating metal mixtures use both DTPA and CaCl2.

Exploring the Use of Species Sensitivity Distributions to Define Protective Limits for the Use of Organic Wastes as Soil Amendments.

The use of organic wastes as soil amendments can be an important measure to improve soil quality and reduce waste accumulation and landfilling. However, the potential contaminant loads of such wastes, can be a source of environmental concern. Consequently, legislation has been developed to regulate the use of these wastes in agricultural soils. However, the regulations only consider chemical parameters, which are insufficient to establish the level of environmental risk. A possible solution is the use of species sensitivity distributions (SSDs), employing ecotoxicological data from test batteries that could be incorporated into legislation. In the present study, 2 different hazardous concentrations affecting 5 and 50% of the soil community (HC5 and HC50, respectively) were determined using ecotoxicological data (effect concentrations, 10 and 50% [EC10 and EC50, respectively]) for 5 different wastes. The results demonstrate that, as expected, current legislative thresholds do not translate to environmental risk/protection and that SSDs may be an important tool allowing the simple inclusion and interpretation of ecotoxicological data from test batteries in legislation. On the other hand, SSDs must be used with caution because there are still doubts about their actual value in risk prediction and about which estimates provide adequate protection. For instance, the use of HC50EC10 values is not recommended; these values overlap with the more conservative HC5EC50 data, highlighting the fact that the use of lower effect concentrations may not always provide the most protective approach. Also, hazardous concentrations need to be calibrated at the field or semifield level, to verify environmental protection in different soils/environments and the adequacy of standard test organisms. Environ Toxicol Chem 2019;38:1569-1576. © 2019 SETAC.

From laboratory to the field: Validating molecular markers of effect in Folsomia candida exposed to a fungicide-based formulation.

Under controlled laboratory conditions, toxicity data tend to be less variable than in more realistic in-field studies and responses may thus differ from those in the natural environment, creating uncertainty. The validation of data under environmental conditions is therefore a major asset in environmental risk assessment of chemicals. The present study aimed to validate the mode of action of a commercial fungicide formulation in the soil invertebrate F. candida, under more realistic exposure scenarios (in-field bioassay), by targeting specific molecular biomarkers retrieved from laboratory experiments. Organisms were exposed in soil cores under minimally controlled field conditions for 4 days to a chlorothalonil fungicide dosage causing 75% reduction of reproduction in a previous laboratory experiment (127 mg a.i. kg-1) and half this concentration (60 mg a.i. kg-1). After exposure, organisms were retrieved and RNA was extracted from each pool of organisms. According to previous laboratorial omics results with the same formulation, ten genes were selected for gene expression analysis by qRT-PCR, corresponding to key genes of affected biological pathways including glutathione metabolism, oxidation-reduction, body morphogenesis, and reproduction. Six of these genes presented a dose-response trend with higher up- or down-regulation with increasing pesticide concentrations. Highly significant correlations between their expression patterns in laboratory and in-field experiments were observed. This work shows that effects of toxicants can be clearly demonstrated in more realistic conditions using validated biomarkers. Our work outlines a set of genes that can be used to assess the early effects of pesticides in a realistic agricultural scenario.

Effects of the neonicotinoids acetamiprid and thiacloprid in their commercial formulations on soil fauna.

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

Organic wastes as soil amendments - Effects assessment towards soil invertebrates.

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