The MetaInvert soil invertebrate genome resource provides insights into below-ground biodiversity and evolution.

Soil invertebrates are among the least understood metazoans on Earth. Thus far, the lack of taxonomically broad and dense genomic resources has made it hard to thoroughly investigate their evolution and ecology. With MetaInvert we provide draft genome assemblies for 232 soil invertebrate species, representing 14 common groups and 94 families. We show that this data substantially extends the taxonomic scope of DNA- or RNA-based taxonomic identification. Moreover, we confirm that theories of genome evolution cannot be generalised across evolutionarily distinct invertebrate groups. The soil invertebrate genomes presented here will support the management of soil biodiversity through molecular monitoring of community composition and function, and the discovery of evolutionary adaptations to the challenges of soil conditions.

Simplification of soil biota communities impairs nutrient recycling and enhances above- and belowground nitrogen losses.

Agriculture is a major source of nutrient pollution, posing a threat to the earth system functioning. Factors determining the nutrient use efficiency of plant-soil systems need to be identified to develop strategies to reduce nutrient losses while ensuring crop productivity. The potential of soil biota to tighten nutrient cycles by improving plant nutrition and reducing soil nutrient losses is still poorly understood. We manipulated soil biota communities in outdoor lysimeters, planted maize, continuously collected leachates, and measured N2 O- and N2 -gas emissions after a fertilization pulse to test whether differences in soil biota communities affected nutrient recycling and N losses. Lysimeters with strongly simplified soil biota communities showed reduced crop N (-20%) and P (-58%) uptake, strongly increased N leaching losses (+65%), and gaseous emissions (+97%) of N2 O and N2 . Soil metagenomic analyses revealed differences in the abundance of genes responsible for nutrient uptake, nitrate reduction, and denitrification that helped explain the observed nutrient losses. Soil biota are major drivers of nutrient cycling and reductions in the diversity or abundance of certain groups (e.g. through land-use intensification) can disrupt nutrient cycling, reduce agricultural productivity and nutrient use efficiency, and exacerbate environmental pollution and global warming.

Cryptic Species in Ecotoxicology.

The advent of genetic methods has led to the discovery of an increasing number of species that previously could not be distinguished from each other on the basis of morphological characteristics. Even though there has been an exponential growth of publications on cryptic species, such species are rarely considered in ecotoxicology. Thus, the particular question of ecological differentiation and the sensitivity of closely related cryptic species is rarely addressed. Tackling this question, however, is of key importance for evolutionary ecology, conservation biology, and, in particular, regulatory ecotoxicology. At the same time, the use of species with (known or unknown) cryptic diversity might be a reason for the lack of reproducibility of ecotoxicological experiments and implies a false extrapolation of the findings. Our critical review includes a database and literature search through which we investigated how many of the species most frequently used in ecotoxicological assessments show evidence of cryptic diversity. We found a high proportion of reports indicating overlooked species diversity, especially in invertebrates. In terrestrial and aquatic realms, at least 67% and 54% of commonly used species, respectively, were identified as cryptic species complexes. The issue is less prominent in vertebrates, in which we found evidence for cryptic species complexes in 27% of aquatic and 6.7% of terrestrial vertebrates. We further exemplified why different evolutionary histories may significantly determine cryptic species' ecology and sensitivity to pollutants. This in turn may have a major impact on the results of ecotoxicological tests and, consequently, the outcome of environmental risk assessments. Finally, we provide a brief guideline on how to deal practically with cryptic diversity in ecotoxicological studies in general and its implementation in risk assessment procedures in particular. Environ Toxicol Chem 2023;42:1889-1914. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Variation in the Chemical Sensitivity of Earthworms from Field Populations to Imidacloprid and Copper.

The chemical risk of pesticides for nontarget soil macroorganisms has mainly been assessed using the compost earthworm Eisenia fetida. However, E. fetida does not occur in agroecosystems, and it is generally less sensitive than other earthworm species. Thus, the extrapolation of its response to pesticides to other earthworm species may lead to uncertainties in risk assessment. Because toxicity data for other earthworms are scarce, we assessed the chemical sensitivity of five species (Allolobophora chlorotica, Aporrectodea caliginosa, Aporrectodea longa, Aporrectodea rosea, and Lumbricus rubellus) from different habitats (forests, wetlands, and grasslands), as well as E. fetida, to imidacloprid and copper in single-species acute toxicity tests. In addition, we examined the relationship between earthworm traits (ecotype and weight), habitat characteristics (ecosystem type and soil pH), and chemical sensitivity. The lower limits of the hazardous concentration affecting 5% (HC5) of species were 178.99 and 0.32 mg active ingredient/kg dry weight for copper and imidacloprid, respectively. Some concentrations that have been measured in European agroecosystems for both pesticides were above the HC5s, indicating toxic risks for these organisms. Furthermore, soil pH from the sampling habitat played a significant role, with earthworms sampled from extremely acidic soils being less sensitive to copper than earthworms from neutral soils. In addition, endogeic earthworms were more sensitive to imidacloprid than epigeic earthworms. This may translate to changes in soil functions such as bioturbation, which is mainly carried out by endogeic earthworms. Our results suggest that risk assessment should include a wider range of earthworms covering different habitats and ecosystem functions to achieve a better protection of the biological functions carried out by these key soil organisms. Environ Toxicol Chem 2023;42:939-947. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Validation of a method to determine transformation of chemicals in anaerobic liquid pig and cattle manure for the OECD test guideline programme.

Manure is widely used as a fertilizer and applied to agricultural land. It may contain highly active chemicals like veterinary medicinal products or biocides, which enter into the environment by this pathway. This is recognized by several regulatory frameworks, however, a detailed method for examining the transformation of chemicals in manure was lacking. This article describes the validation of a method for studying the anaerobic transformation of chemicals in pig and cattle liquid manure. Different steps are covered with an emphasis on the validation ring test and the OECD (Organisation for Economic Cooperation and Development) process that led to the recent adoption of the method as OECD Test Guideline (TG) 320.

Shotgun metagenomics of soil invertebrate communities reflects taxonomy, biomass, and reference genome properties.

Metagenomics - shotgun sequencing of all DNA fragments from a community DNA extract - is routinely used to describe the composition, structure, and function of microorganism communities. Advances in DNA sequencing and the availability of genome databases increasingly allow the use of shotgun metagenomics on eukaryotic communities. Metagenomics offers major advances in the recovery of biomass relationships in a sample, in comparison to taxonomic marker gene-based approaches (metabarcoding). However, little is known about the factors which influence metagenomics data from eukaryotic communities, such as differences among organism groups, the properties of reference genomes, and genome assemblies.We evaluated how shotgun metagenomics records composition and biomass in artificial soil invertebrate communities at different sequencing efforts. We generated mock communities of controlled biomass ratios from 28 species from all major soil mesofauna groups: mites, springtails, nematodes, tardigrades, and potworms. We shotgun sequenced these communities and taxonomically assigned them with a database of over 270 soil invertebrate genomes.We recovered over 95% of the species, and observed relatively high false-positive detection rates. We found strong differences in reads assigned to different taxa, with some groups (e.g., springtails) consistently attracting more hits than others (e.g., enchytraeids). Original biomass could be predicted from read counts after considering these taxon-specific differences. Species with larger genomes, and with more complete assemblies, consistently attracted more reads than species with smaller genomes. The GC content of the genome assemblies had no effect on the biomass-read relationships. Results were similar among different sequencing efforts.The results show considerable differences in taxon recovery and taxon specificity of biomass recovery from metagenomic sequence data. The properties of reference genomes and genome assemblies also influence biomass recovery, and they should be considered in metagenomic studies of eukaryotes. We show that low- and high-sequencing efforts yield similar results, suggesting high cost-efficiency of metagenomics for eukaryotic communities. We provide a brief roadmap for investigating factors which influence metagenomics-based eukaryotic community reconstructions. Understanding these factors is timely as accessibility of DNA sequencing and momentum for reference genomes projects show a future where the taxonomic assignment of DNA from any community sample becomes a reality.

Holistic evaluation of long-term earthworm field studies with a fungicide.

Plant protection products to be placed on the market in the European Union need to meet rigorous safety criteria including the testing of lumbricid earthworms, the functionally most important soil organism group in Central European agricultural ecosystems. To address uncertainties and investigate the potential long-term in-crop effects of the fungicide Cantus® containing 50% boscalid as an active substance, a series of standardized earthworm field studies with an overall duration of 5 years per study program was carried out in four German agricultural fields under realistic crop rotation conditions. A two-step approach was chosen to analyze the potential overall long-term effects on earthworms in agricultural fields: (i) an assessment of the earthworm abundance development in the course of the four study programs in relation to the determined actual content of boscalid in soil and (ii) an effect size meta-analysis of earthworm abundance 1 year after treatment for each consecutive year and study program. Measured boscalid concentrations in the soil after multiple applications were well above the maximum boscalid residues observed in agricultural soils across Central Europe. There were isolated statistically significant reductions of earthworm abundance for some species and groups at some time points during the studies, but no consistent relationship to the Cantus® treatments was observed. These results were supported by the meta-analysis, indicating no adverse effects on earthworm populations. Therefore, fluctuations of abundance reflect the natural variation of the populations rather than a concentration-related response. Based on this comprehensive analysis, we conclude that there is no application rate-related effect of the 5-year use of Cantus® on the development of the earthworm communities. The four study programs, paired with a comprehensive evaluation, directly address the concerns about the potential long-term effects of boscalid on earthworms in the field and suggest that multiyear applications do not adversely affect earthworm populations. Integr Environ Assess Manag 2022;18:1399-1413. © 2021 ECT Oekotoxikologie GmbH and BASF SE. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Application of the Closure Principle Computational Approach Test to Assess Ecotoxicological Field Studies: Comparative Analysis Using Earthworm Field Test Abundance Data.

Field studies to determine the effects of chemicals on earthworm communities are generally conducted according to International Organization for Standardization standard 11268-3 (and later comments). However, statistical test procedures suggested in the guideline are frequently criticized, mainly for 2 reasons: 1) Earthworm abundances are count data and often do not fulfill requirements for multiple t tests (normal distribution and homogeneity of variance), and 2) the resulting toxicity metrics of multiple testing procedures (no/lowest-observed-effect concentrations [NOEC/LOEC]) fail to adequately detect the actual level of effects. Recently, a new method to overcome these shortcomings was presented by the introduction of the closure principle computational approach test (CPCAT). We applied this statistical method to assess chemical effects on abundance in a large dataset of 26 earthworm field studies (with up to 3 test chemical application rates) and an additional extended study with 6 application rates. A comparative analysis was provided considering results of well-established multiple testing approaches (Dunnett's test) with particular consideration of the degree of overdispersion found in these data. It was shown that the CPCAT detects substantially more effects in earthworm field tests as statistically significant than standard t test approaches. This lowered the LOEC/NOEC for many chemical treatments to control comparisons. As a consequence, the statistically detected NOECs/LOECs were often set at lower percentage deviations between control and chemical treatment. This is the first time the performance of the CPCAT has been assessed within a comprehensive analysis of earthworm field study data. Environ Toxicol Chem 2021;40:1750-1760. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Potential of Eucalyptus globulus for the phytoremediation of metals in a Moroccan iron mine soil-a case study.

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

Application of a standard risk assessment scheme to a North Africa contaminated site (Sfax, Tunisia) -Tier 1.

Phosphorus is a critical element to agriculture, consequently global phosphate rock demand will remain rising to feed a growing world population. The beneficiation of phosphorous ore gives rise to several tons of a waste by-product [phosphogypsum (PG)] which valorisation is limited, within other reasons, by the risks posed to environment and human health. Although threatening, the accumulation in stacks is the only procedure so far practiced by several countries as a means to get rid of this industrial externality. As part of a NATO Science for Peace Project (SfP 983311) this study describes the application of an environmental risk assessment (ERA) framework, to assess the risks posed by a PG stack to the surrounding soils, in Sfax, Republic of Tunisia. The ERA followed a weight of evidence approach, supported by two lines of evidence (LoE): the chemical (ChemLoE) and the ecotoxicological (EcotoxLoE). Integrated risks point for risk values greater than 0.5 in soils collected in PG stack surrounding area. Soil salinization, has likely contributed to the exacerbation of risks, as well as to the lack of consistency between both LoEs. This study highlights the need of rethinking the weight given to each LoE in ERA, in areas where soil salinization is a reality.

Soil Biodiversity: State-of-the-Art and Possible Implementation in Chemical Risk Assessment.

Protecting the structure and functioning of soil ecosystems is one of the central aims of current regulations of chemicals. This is, for instance, shown by the emphasis on the protection of key drivers and ecosystem services as proposed in the protection goal options for soil organisms by the European Food Safety Authority (EFSA). Such targets require insight into soil biodiversity, its role in the functioning of ecosystems, and the way it responds to stress. Also required are tools and methodologies for properly assessing biodiversity. To address these issues, the Society of Environmental Toxicology and Chemistry (SETAC) Europe 14th Special Science Symposium (SESSS14) was held 19 to 20 November 2019 in Brussels, Belgium. The central aim of the SESSS14 was to provide information on how to include soil biodiversity and soil functions as protection goal options in the risk assessment and quantification of the effects of chemicals and other stressors (including their respective regulations). This paper is based on the presentations and discussions at the SESSS14 and will give a brief update on the scientific state-of-the art on soil biodiversity, novel scientific developments, experimental and modeling approaches, as well as case studies. It will also discuss how these approaches could inform future risk assessment of chemicals and other stressors in the regulatory context of protecting soil ecosystems. Integr Environ Assess Manag 2021;17:541-551. © 2020 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Acute and chronic toxicity of the fungicide carbendazim to the earthworm Eisenia fetida under tropical versus temperate laboratory conditions.

Research efforts into the potential side-effects of pesticides on beneficial organisms have focused on temperate test species and conditions. There is thus a need for studies into the ecotoxicity of a vaster range of pesticides under tropical conditions. The present study therefore aimed to compare the acute and chronic toxicity of the fungicide carbendazim to the earthworm Eisenia fetida under tropical and temperate conditions. To this end, laboratory toxicity tests were conducted with a tropical and European strain of E. fetida, using different artificial (OECD and TAS) and natural (LUFA and TNS) soils, and under different test temperatures (20 °C and 28 °C). In the acute lethality tests with artificial soils, the tropical strain of E. fetida was three to four order of magnitude less sensitive than the European strain, which is ascribed to the higher test temperature and (hence) higher microbial activity/pesticide degradation. The tropical strain was particularly sensitive in the tropical natural soil, which was attributed to the low pH (3.9) of this soil. The chronic toxicity tests overall also showed a lower sensitivity of the tropical strain on reproduction. These findings thus support the use of toxicity data generated under temperate conditions in tropical pesticide effect assessments. However, intensive agricultural practices in the tropics may dictate that exposure levels (and hence potentially also risks) are higher.

Commercial glyphosate-based herbicides effects on springtails (Collembola) differ from those of their respective active ingredients and vary with soil organic matter content.

Glyphosate-based herbicides (GBH) are currently the most widely used agrochemicals for weed control. Environmental risk assessments (ERA) on nontarget organisms mostly consider the active ingredients (AIs) of these herbicides, while much less is known on effects of commercial GBH formulations that are actually applied in the field. Moreover, it is largely unknown to what extent different soil characteristics alter potential side effects of herbicides. We conducted a greenhouse experiment growing a model weed population of Amaranthus retroflexus in arable field soil with either 3.0 or 4.1% soil organic matter (SOM) content and treated these weeds either with GBHs (Roundup LB Plus, Touchdown Quattro, Roundup PowerFlex) or their respective AIs (isopropylammonium, diammonium or potassium salts of glyphosate) at recommended dosages. Control pots were mechanically weeded. Nontarget effects were assessed on the surface activity of the springtail species Sminthurinus niger (pitfall trapping) and litter decomposition in the soil (teabag approach). Both GBHs and AIs increased the surface activity of springtails compared to control pots; springtail activity was higher under GBHs than under corresponding AIs. Stimulation of springtail activity was much higher in soil with higher SOM content than with low SOM content (significant treatment x SOM interaction). Litter decomposition was unaffected by GBHs, AIs or SOM levels. We suggest that ERAs for pesticides should be performed with actually applied herbicides rather than only on AIs and should also consider influences of different soil properties.

Effects of waste materials on Caenorhabditis elegans (Nematoda) using the ISO standard soil toxicity test.

The ecotoxicological characterization of waste according to the European Waste List (HP14) is part of its hazard classification, which is based on 15 different hazardous properties and should include toxicity tests representing the aquatic and terrestrial compartment. Besides established soil toxicity tests with bacteria, plants, and earthworms, the standardized test with the nematode Caenorhabditis elegans (ISO 10872:2010) is suitable for testing soils and wastes, however, has never been validated for the purpose of waste toxicity assessment. Therefore, 23 different waste samples were tested for their toxicity on growth and reproduction of C. elegans to validate the suitability of ISO 10872 (ISO 2010) for assessing the ecotoxicological hazard of waste and to compare the results with those of other ecotoxicological test systems. C. elegans showed a comparable response to the waste samples as the earthworm avoidance test, however, with single samples where the nematodes indicated a higher toxicity than the E. fetida, thus, providing non-redundant information to the ecotoxicological hazard assessment of wastes. Also due to the short duration (4 days) and small-scale test set-up, the soil toxicity test with C. elegans turned out to be a valuable addition to already existing test batteries, for assessing the ecotoxicity of wastes in the soil compartment.

Environmental risk assessment of pesticides in tropical terrestrial ecosystems: Test procedures, current status and future perspectives.

Despite the increasing use of pesticides in tropical countries, research and legislative efforts have focused on their temperate counterparts. This paper presents a review of the literature on environmental risk assessment of pesticides for tropical terrestrial agroecosystems. It aims at evaluating potential differences in pesticide risk between temperate and tropical regions as well as to highlight research needs in the latter. Peculiarities of pesticide risks in tropical terrestrial agroecosystems are discussed in subsections 1) agricultural practices; 2) research efforts; 3) fate and exposure; 4) toxicity testing methods; and 5) sensitivity. The intensive and often inadequate pesticide application practices in tropical areas are likely to result in a relatively greater pesticide exposure in edge-of-field water bodies. Since pesticide fate may be different under tropical conditions, tropical scenarios for models estimating predicted environmental pesticide concentrations should be developed. Sensitivity comparisons do not indicate a consistent similar, greater or lower relative sensitivity of tropical soil organisms as compared to temperate organisms. However, several methods and procedures for application in the tropics need to be developed, which include: 1) identifying and collecting natural soils to be used as reference test substrates in tests; 2) identifying and discerning the range of sensitivity of native test species to soil contaminants; 3) developing test guidelines applicable to tropical/subtropical conditions; and 4) developing methods and procedures for higher tier testing for full development and implementation of environmental risk assessment schemes.

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.

A new ecotoxicological test method for genetically modified plants and other stressors in soil with the black fungus gnat Bradysia impatiens (Diptera): current status of test development and dietary effects of azadirachtin on larval development and emergence rate.

BACKGROUND: Few suitable and standardized test methods are currently available to test the effects of genetically modified plants (GMP) on non-target organisms. To fill this gap and improve ecotoxicological testing for GMP, we developed a new soil ecotoxicological test method using sciarid larvae as test organisms. RESULTS: Bradysia impatiens was identified as a candidate species. Species of the genus Bradysia occur in high numbers in European agroecosystems and B. impatiens can be reared in the laboratory in continuous culture. A functional basic test design was successfully developed. Newly hatched larvae were used as the initial life stage to cover most of the life cycle of the species during the test. Azadirachtin was identified as a suitable reference substance. In several tests, the effects of this substance on development time and emergence rate varied for different temperatures and test substrates. The toxicity was higher at 25 °C compared to 20 °C and in tropical artificial soil compared to coconut fiber substrate. CONCLUSIONS AND OUTLOOK: Results suggest that the developed test system is suitable to enter a full standardization process, e.g., via the Organisation for Economic Co-operation and Development. Such a standardization would not only assist the risk assessment of GMP, but could include other stressors such as systemic pesticides or veterinary pharmaceuticals reaching the soil, e.g., via spreading manure. The use of sciarid flies as test organisms supports recommendations of EFSA, which stressed the ecological role of flies and encouraged including Diptera into test batteries.

Toward sustainable environmental quality: Priority research questions for Europe.

The United Nations' Sustainable Development Goals have been established to end poverty, protect the planet, and ensure prosperity for all. Delivery of the Sustainable Development Goals will require a healthy and productive environment. An understanding of the impacts of chemicals which can negatively impact environmental health is therefore essential to the delivery of the Sustainable Development Goals. However, current research on and regulation of chemicals in the environment tend to take a simplistic view and do not account for the complexity of the real world, which inhibits the way we manage chemicals. There is therefore an urgent need for a step change in the way we study and communicate the impacts and control of chemicals in the natural environment. To do this requires the major research questions to be identified so that resources are focused on questions that really matter. We present the findings of a horizon-scanning exercise to identify research priorities of the European environmental science community around chemicals in the environment. Using the key questions approach, we identified 22 questions of priority. These questions covered overarching questions about which chemicals we should be most concerned about and where, impacts of global megatrends, protection goals, and sustainability of chemicals; the development and parameterization of assessment and management frameworks; and mechanisms to maximize the impact of the research. The research questions identified provide a first-step in the path forward for the research, regulatory, and business communities to better assess and manage chemicals in the natural environment. Environ Toxicol Chem 2018;37:2281-2295. © 2018 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

A bacterium-based contact assay for evaluating the quality of solid samples-Results from an international ring-test.

The contact assay measuring the inhibition of Arthrobacter globiformis dehydrogenase activity as an endpoint to evaluate the toxicity of solid samples was tested in an international ring-test to validate its performance for ISO standardization (ISO/CD 18187). This work reports the results of the ring-test involving 9 laboratories from six countries. At least 8 valid data sets were obtained for each sample and more than three quarters of the participants attained the validity criteria defined in the standard. The coefficient of variation within (CVr) and between (CVR) laboratories was generally on average <15% and <30% for negative and positive controls, respectively. Regarding solid samples, the laboratories provided a similar ranking of the samples based on their toxicity, despite some variation in the LOEC values. The logarithmic within-lab standard deviation <0.50 for soils and <0.25 for wastes evidenced a good repeatability. The between-lab variability assessed by a CVR <30%, minimum-maximum factor <4 and a reproducibility standard deviation (SDR) <0.13 for a great part of the solid samples, confirmed the test reproducibility. Overall, this assay proved to be robust, sensitive and feasible for routine use towards the quality assessment of soils and wastes.

Standard methods for the assessment of structural and functional diversity of soil organisms: A review.

The lack of standardized methods to study soil organisms prevents comparisons across data sets and the development of new global and regional experiments and assessments. Moreover, standardized methods are needed to evaluate the impact of anthropogenic stressors, such as chemicals, on soil organism communities in the regulatory context. The goal of this contribution is to summarize current methodological approaches to measure structural and functional diversity of soil organisms, and to identify gaps and methodological improvements so as to cross data sets generated worldwide. This is urgently needed because several currently ongoing regional and global soil biodiversity studies are not coordinated with one another in terms of methodology, including database development. Therefore, we evaluated the standard methods to sample, identify, determine, and assess soil organisms currently applied or proposed, using well-accepted criteria such as ecological relevance; practicability of usage in terms of resources, time, and costs; and the level of standardization. Methods addressing both the structure and the functions of soil organisms (populations or communities) are included, with a special focus on new molecular methods based on nucleic acid extraction and further analyses by polymerase chain reaction (PCR)-based approaches for microorganisms and invertebrates. We particularly highlight the activities of the Technical Committee (TC) 190 of the International Organization for Standardization (ISO) because ISO guidelines are legally accredited by many national or international authorities when they put conservation laws and regulations into practice. Finally, we propose detailed recommendations regarding gaps in the available set of standards, in order to identify a list of new methods to be standardized. We propose to organize this whole process under the Global Soil Biodiversity Initiative (GSBI) in order to ensure a truly global approach for the assessment of soil biodiversity. Integr Environ Assess Manag 2018;14:463-479. © 2018 SETAC.