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The efficient management of soil represents a mission of vital importance for meeting the continuously increasing agricultural demand in a sustainable way. Decades of research identified in the biotechnological potential of soil microorganisms an always more practicable channel for achieving these goals. Due to the complexity of soil microbial communities and their tight connection to soil characteristics, it is still difficult to define universal strategies for an efficient and sustainable agroecosystem management. We here propose a new framework for the assessment of the impact of agricultural practices in the agroecosystem that revolves around the concept of microbial community recovery. This assessment is based on the selection of (i) a representative temporal interval, (ii) a representative agricultural system and (iii) monitoring tools able to assess the expression levels of microbial functionality in soil. This approach can be especially valuable for evaluating the effects of agrochemicals and other agronomical amendments (of different nature: biological, physical, chemical) on the soil microbiota. In the same way precision-medicine tries to tailor drugs on an always smaller subset of patients' characteristics, a new generation of agrochemicals can be developed and tested considering soil characteristics in order to minimize their off-target effects. What remains central in this paradigm is the promotion of Soil Health maintenance practices. As for healthy humans, a healthy soil is more resilient and tolerates treatments and stresses better while recovering more quickly.

Ecological risk assessment (ERA) has followed a taxonomy-based approach, making the assumption that related species will show similar sensitivity to toxicants, and using safety factors or species sensitivity distributions to extrapolate from tested to untested species. In ecology it has become apparent that taxonomic approaches may have limitations for the description and understanding of species assemblages in nature. Therefore it has been proposed that the inclusion of species traits in ERA could provide a useful and alternative description of the systems under investigation. At the same time, there is a growing recognition that the use of mechanistic approaches in ERA, including conceptual and quantitative models, may improve predictive and extrapolative power. Purposefully linking traits with mechanistic effect models could add value to taxonomy-based ERA by improving our understanding of how structural and functional system facets may facilitate inter-species extrapolation. Here, we explore whether and in what ways traits can be linked purposefully to mechanistic effect models to predict intrinsic sensitivity using available data on the acute sensitivity and toxicokinetics of a range of freshwater arthropods exposed to chlorpyrifos. The results of a quantitative linking of seven different endpoints and twelve traits demonstrate that while quantitative links between traits and/or trait combinations and process based (toxicokinetic) model parameters can be established, the use of simple traits to predict classical sensitivity endpoints yields little insight. Remarkably, neither of the standard sensitivity values, i.e. the LC(50) or EC(50), showed a strong correlation with traits. Future research in this area should include a quantitative linking of toxicodynamic parameter estimations and physiological traits, and requires further consideration of how mechanistic trait-process/parameter links can be used for prediction of intrinsic sensitivity across species for different substances in ERA.

In this chapter we review the ecotoxicology of the synthetic pyrethroids (SPs). SPs are potent, broad-spectrum insecticides. Their effects on a wide range of nontarget species have been broadly studied, and there is an extensive database available to evaluate their effects. SPs are highly toxic to fish and aquatic invertebrates in the laboratory, but effects in the field are mitigated by rapid dissipation and degradation. Due to their highly lipophilic nature, SPs partition extensively into sediments. Recent studies have shown that toxicity in sediment can be predicted on the basis of equilibrium partitioning, and whilst other factors can influence this, organic carbon content is a key determining variable. At present for SPs, there is no clear evidence for adverse population-relevant effects with an underlying endocrine mode of action. SPs have been studied intensively in aquatic field studies, and their effects under field conditions are mitigated from those measured in the laboratory by their rapid dissipation and degradation. Studies with a range of test systems have shown consistent aquatic field endpoints across a variety of geographies and trophic states. SPs are also highly toxic to bees and other nontarget arthropods in the laboratory. These effects are mitigated in the field through repellency and dissipation of residues, and recovery from any adverse effects tends to be rapid.

A key challenge in ecotoxicology is to assess the potential risks of chemicals to the wide range of species in the environment on the basis of laboratory toxicity data derived from a limited number of species. These species are then assumed to be suitable surrogates for a wider class of related taxa. For example, Daphnia spp. are used as the indicator species for freshwater aquatic invertebrates. Extrapolation from these datasets to natural communities poses a challenge because the extent to which test species are representative of their various taxonomic groups is often largely unknown, and different taxonomic groups and chemicals are variously represented in the available datasets. Moreover, it has been recognized that physiological and ecological factors can each be powerful determinants of vulnerability to chemical stress, thus differentially influencing toxicant effects at the population and community level. Recently it was proposed that detailed study of species traits might eventually permit better understanding, and thus prediction, of the potential for adverse effects of chemicals to a wider range of organisms than those amenable for study in the laboratory. This line of inquiry stems in part from the ecology literature, in which species traits are being used for improved understanding of how communities are constructed, as well as how communities might respond to, and recover from, disturbance (see other articles in this issue). In the present work, we develop a framework for the application of traits-based assessment. The framework is based on the population vulnerability conceptual model of Van Straalen in which vulnerability is determined by traits that can be grouped into 3 major categories, i.e., external exposure, intrinsic sensitivity, and population sustainability. Within each of these major categories, we evaluate specific traits as well as how they could contribute to the assessment of the potential effects of a toxicant on an organism. We then develop an example considering bioavailability to explore how traits could be used mechanistically to estimate vulnerability. A preliminary inventory of traits for use in ecotoxicology is included; this also identifies the availability of data to quantify those traits, in addition to an indication of the strength of linkage between the trait and the affected process. Finally, we propose a way forward for the further development of traits-based approaches in ecotoxicology., (Copyright © 2011 SETAC.)

Recent advances in modeling the processes of the toxicity of chemicals-toxicokinetics (TK) and toxicodynamics (TD)-are improving environmental risk assessment (ERA) through prediction of effects from time-varying exposure. This has been achieved by linking chemical fate and toxicological effects mechanistically, based on internal concentrations, through the tissue residue approach. However, certain questions remain: for example, how do TK and TD differ among species and how does this relate to differences in species sensitivity? In a series of experiments, we studied the TK of [(14)C]chlorpyrifos in 15 freshwater arthropod species, two of which were studied in juvenile and adult life stages. Uptake (k(in)) and elimination (k(out)) rate constants were fitted using a one-compartment single first-order kinetic model. The application of two complementary parameter estimation methods facilitated the calculation of bioconcentration factors (BCF) with prediction intervals and 95% depuration times (t(95)) for all tested species. Extremely slow elimination was observed in some species as well as high overall variation in k(in), k(out), BCF, and t(95) across the tested aquatic arthropod species. This variation has implications for the development of TKTD approaches in ERA, including assessing fluctuating exposure concentrations and the interpretation of observed toxicity responses in the laboratory and in the field., (Environ. Toxicol. Chem. 2010;29:2225-2234. © 2010 SETAC.)

Under typical agricultural use of an insecticide, it is likely that only part of an edge-of-field drainage ditch will be directly contaminated by spray drift. The response, including recovery, of aquatic macroinvertebrates in sprayed ditch sections may be affected by immigration of organisms from adjacent nonsprayed ditch sections, but also the population dynamics in nonsprayed sections (refuges) may be affected by nearby contaminated patches (known as action at a distance). Experimental ditches were used to study the influence of the presence of nearby refuges on the responses of macroinvertebrates in ditch sections directly sprayed with the insecticide lufenuron, and vice versa. The treatment regimes differed in the proportion of the ditch (0, 33, 67, and 100% of surface area) that was sprayed to reach a lufenuron concentration of 3 microg/L in the water column of the sprayed ditch section. In sprayed ditch sections, clear treatment-related effects were observed for adult midges in the emergence traps and for aquatic arthropods (mainly juveniles) in the artificial substrate/sweep net samples. The extent in magnitude and duration of effects in sprayed ditch sections was overall larger when a larger proportion of the ditch was sprayed and/or the distance to the refuge was larger. In nonsprayed ditch sections of partially treated ditches, treatment-related effects were absent or minor for macroinvertebrates that predominantly dwell on or in the sediment compartment, particularly at a larger distance from the sprayed ditch sections. More mobile arthropods that predominantly dwell in the water column showed clear treatment-related effects in the nonsprayed ditch sections as well, but action at a distance was smaller if a smaller proportion of ditch was treated., (Copyright 2010 SETAC.)

This study evaluates the effects of the triazine herbicide simazine in an outdoor pond microcosm test system that contained two submerged rooted species (Myriophyllum spicatum and Elodea canadensis) and two emergent rooted species (Persicaria amphibia and Glyceria maxima) over a period of 84 days. Simazine was applied to the microcosms at nominal concentrations of 0.05, 0.5 and 5 mg/L. General biological endpoints and physiological endpoints were used to evaluate herbicide toxicity on macrophytes and the algae developing naturally in the system. Concentration-related responses of macrophytes and algae were obtained for the endpoints selected, resulting in a no observed ecologically adverse effect concentration (NOEAEC) at simazine concentrations of 0.05 mg active ingredient/L after 84 days. E. canadensis was the most negatively affected species based on length increase, which was consistently a very sensitive parameter for all macrophytes. The experimental design presented might constitute a suitable alternative to conventional laboratory single-species testing., (Copyright (c) 2009 Elsevier Ltd. All rights reserved.)

The long-term response, including recovery, of aquatic macroinvertebrates to short-term insecticide exposure may be affected by the presence of uncontaminated refuges in the stressed ecosystem. Experimental ditches were used to study the influence of non-sprayed ditch sections regarding the ecotoxicological effects on and the recovery of macroinvertebrates following treatment with the insecticide lufenuron. The treatment regimes differed in the proportion of the ditch (0, 33, 67, and 100% of surface area) that was sprayed to reach a lufenuron concentration of 3 microg/L in the water column of the sprayed ditch section. The magnitude and duration of effects on macroinvertebrates, and on arthropods in particular, were higher when a larger proportion of the ditch was treated. Initially, more pronounced responses were observed for bivoltine and multivoltine insects and macrocrustaceans than for univoltine and semivoltine arthropods. Most macroinvertebrate arthropods showed delayed responses, with maximum treatment-related effects observed two to six weeks after lufenuron application. This latency of effects can be explained by the mode of action of lufenuron, involving inhibition of chitin synthesis, which affects arthropod molting and metamorphosis. The observed effects were short-lived only in those ditches where 33% of the surface area was sprayed. In the ditches where 67 and 100% of the surface area was sprayed, some insects and macrocrustaceans showed long-term effects. In the 100% sprayed ditches in particular, the treatment-related reduction in arthropods resulted in indirect effects, such as an increase in snails, and later in an increase in the ephemeropteran Cloeon dipterum, probably because of an increase in periphyton, and release from competition and predation. Effects that are most likely indirect also were observed for Oligochaeta, Hirudinea, and the flatworm Mesostoma sp.

Outdoor experimental ditches were used to evaluate the influence of untreated refuges on the recovery of zooplankton communities following treatment with the fast-dissipating insecticide lufenuron. Each experimental ditch was divided into three sections of the same surface area. The treatments differed in the proportion of ditch (0, 33, 67, and 100% of the surface area) to which the insecticide was applied at the same nominal treatment (3 mug/L). During the first week postapplication, a barrier was placed between treated and untreated ditch sections. The untreated sections were included to provide a source of organisms for recovery of affected zooplankton populations in the treated sections of the ditch after the removal of the barrier. Cyclopoida were the most affected by lufenuron treatment, followed by Daphnia gr. galeata. These and other direct effects of treatment on larvae of the phantom midge Chaoborus spp. resulted in clear indirect effects on populations of Calanoida, Ceriodaphnia, and Rotifera. Overall, faster recovery of the zooplankton community was observed in the treated sections of ditches that were sprayed for a smaller proportion of their surface area. Nevertheless, individual zooplankton populations showed considerable differences in rate of recovery. Cyclopoida showed a relatively slow rate of recovery even in the partially treated ditches. Daphnia gr. galeata recovered more rapidly in treated ditch sections in the presence of unsprayed ditch sections, illustrating the potential influence of unexposed refuges. Furthermore, the presence of refuges most likely dampened the magnitude and duration of indirect effects in the ditches treated with lufenuron.

The concept of an organism's niche is central to ecological theory, but an operational definition is needed that allows both its experimental delineation and interpretation of field distributions of the species. Here we use population growth rate (hereafter, pgr) to define the niche as the set of points in niche space where pgr > 0. If there are just two axes to the niche space, their relationship to pgr can be pictured as a contour map in which pgr varies along the axes in the same way that the height of land above sea level varies with latitude and longitude. In laboratory experiments we measured the pgr of Daphnia magna over a grid of values of pH and Ca2+, and so defined its "laboratory niche" in pH-Ca2+ space. The position of the laboratory niche boundary suggests that population persistence is only possible above 0.5 mg Ca2+/L and between pH 5.75 and pH 9, though more Ca2+ is needed at lower pH values. To see how well the measured niche predicts the field distribution of D. magna, we examined relevant field data from 422 sites in England and Wales. Of the 58 colonized water bodies, 56 lay within the laboratory niche. Very few of the sites near the niche boundary were colonized, probably because pgr there is so low that populations are vulnerable to extinction by other factors. Our study shows how the niche can be quantified and used to predict field distributions successfully.

DNA microarrays can be used to measure environmental stress responses. If they are to be predictive of environmental impact, we need to determine if altered gene expression translates into negative impacts on individuals and populations. A large cDNA microarray (14000 spots) was created to measure molecular stress responses to cadmium in Daphnia magna,the mostwidely used aquatic indicator species, and relate responses to population growth rate (pgr). We used the array to detect differences in the transcription of genes in juvenile D. magna (24 h old) after 24 h exposure to a control and three cadmium concentrations (6, 20, and 37 microg Cd2+ L(-1)). Stress responses at the population level were estimated following a further 8 days exposure. Pgr was approximately linear negative with increasing cadmium concentration over this range. The microarray profile of gene expression in response to acute cadmium exposure begins to provide an overview of the molecular responses of D. magna, especially in relation to growth and development. Of the responding genes, 29% were involved with metabolism including carbohydrate, fat and peptide metabolism, and energy production, 31% were involved with transcription/translation, while 40% of responding genes were associated with cellular processes like growth and moulting, ion transport, and general stress responses (which included oxidative stress). Our production and application of a large Daphnia magna microarray has shown that measured gene responses can be logically linked to the impact of a toxicant such as cadmium on somatic growth and development, and consequently pgr.

Background: Ibuprofen and other nonsteroidal anti-inflammatory drugs have been designed to interrupt eicosanoid metabolism in mammals, but little is known of how they affect nontarget organisms. Here we report a systems biology study that simultaneously describes the transcriptomic and phenotypic stress responses of the model crustacean Daphnia magna after exposure to ibuprofen., Results: Our findings reveal intriguing similarities in the mode of action of ibuprofen between vertebrates and invertebrates, and they suggest that ibuprofen has a targeted impact on reproduction at the molecular, organismal, and population level in daphnids. Microarray expression and temporal real-time quantitative PCR profiles of key genes suggest early ibuprofen interruption of crustacean eicosanoid metabolism, which appears to disrupt signal transduction affecting juvenile hormone metabolism and oogenesis., Conclusion: Combining molecular and organismal stress responses provides a guide to possible chronic consequences of environmental stress for population health. This could improve current environmental risk assessment by providing an early indication of the need for higher tier testing. Our study demonstrates the advantages of a systems approach to stress ecology, in which Daphnia will probably play a major role.

The non-steroidal anti-inflammatory drug (NSAID) ibuprofen (IB) is a widely used pharmaceutical that can be found in several freshwater ecosystems. Acute toxicity studies with Daphnia magna suggest that the 48h EC(50) (immobilisation) is 10-100 mgIBl(-1). However, there are currently no chronic IB toxicity data on arthropod populations, and the aquatic life impacts of such analgesic drugs are still undefined. We performed a 14-day exposure of D. magna to IB as a model compound (concentration range: 0, 20, 40 and 80 mgIBl(-1)) measuring chronic effects on life history traits and population performance. Population growth rate was significantly reduced at all IB concentrations, although survival was only affected at 80 mgIBl(-1). Reproduction, however, was affected at lower concentrations of IB (14-day EC(50) of 13.4 mgIBl(-1)), and was completely inhibited at the highest test concentration. The results from this study indicate that the long-term crustacean population consequences of a chronic IB exposure at environmentally realistic concentrations (ngl(-1) to microgl(-1)) would most likely be of minor importance. We discuss our results in relation to recent genomic studies, which suggest that the potential mechanism of toxicity in Daphnia is similar to the mode of action in mammals, where IB inhibits eicosanoid biosynthesis.

A new test design for the non-axenic submergent aquatic macrophytes Elodea canadensis Michx. and Myriophyllum spicatum L. has been developed for potential use in herbicide toxicity testing. For the non-axenic cultures, the best growth conditions were observed in the Elendt-M4 medium in which no growth of algae or bacteria was observed. Cuttings were placed in beakers containing only the artificial M4 medium or were planted in small beakers containing OECD (Organisation for Economic Cooperation and Development) sediment (5% peat, 75% sand, 20% kaolinite), which were then placed in larger vessels with the M4 medium. The plants were observed for main and secondary shoot length, biomass and root formation within 2-3 weeks of planting. Growth rates were calculated for total plant length and biomass. The variance between the replicates was low throughout the experiment [coefficient of variation (CV) < 26% for total plant length, and between 16 and 40% for biomass]. Relative growth rates based on total plant length were determined as 0.028 and 0.050 per day for M. spicatum in the systems containing M4 medium only and medium plus sediment respectively. Similar results were observed for E. canadensis, with relative growth rates of 0.26 and 0.073 per day in the two test systems. The root-shoot ratio at harvest was greater by a factor of 2-3 for E. canadensis in the M4 medium than in the system containing sediment. However, comparable ratios were observed for M. spicatum in the two test systems. Both growth in total plant length and growth in biomass of the two species have potential as measures of toxicity.

Background: Transcriptomic techniques are now being applied in ecotoxicology and toxicology to measure the impact of stressors and develop understanding of mechanisms of toxicity. Microarray technology in particular offers the potential to measure thousands of gene responses simultaneously. However, it is important that microarrays responses should be validated, at least initially, using real-time quantitative polymerase chain reaction (QPCR). The accurate measurement of target gene expression requires normalisation to an invariant internal control e.g., total RNA or reference genes. Reference genes are preferable, as they control for variation inherent in the cDNA synthesis and PCR. However, reference gene expression can vary between tissues and experimental conditions, which makes it crucial to validate them prior to application., Results: We evaluated 10 candidate reference genes for QPCR in Daphnia magna following a 24 h exposure to the non-steroidal anti-inflammatory drug (NSAID) ibuprofen (IB) at 0, 20, 40 and 80 mg IB l-1. Six of the 10 candidates appeared suitable for use as reference genes. As a robust approach, we used a combination normalisation factor (NF), calculated using the geNorm application, based on the geometric mean of three selected reference genes: glyceraldehyde-3-phosphate dehydrogenase, ubiquitin conjugating enzyme and actin. The effects of normalisation are illustrated using as target gene leukotriene B4 12-hydroxydehydrogenase (Ltb4dh), which was up-regulated following 24 h exposure to 63-81 mg IB l-1., Conclusions: As anticipated, use of the NF clarified the response of Ltb4dh in daphnids exposed to sublethal levels of ibuprofen. Our findings emphasise the importance in toxicogenomics of finding and applying invariant internal QPCR control(s) relevant to the study conditions.

Outdoor aquatic ditch mesocosms were treated with a range of pesticides to simulate various spray drift rates resulting from a typical crop protection program used in the cultivation of potatoes in The Netherlands. The main experimental aims of the present study were to provide information on the fate and ecological effects of drift of the pesticides into surface water and to evaluate the effectiveness of drift-reduction measures in mitigating risks. The pesticides selected and the dosage, frequency, and timing of application were based on normal agricultural practices in the potato crop. Applications of prosulfocarb, metribuzin (both herbicides), lambda-cyhalothrin (insecticide), chlorothalonil, and fluazinam (both fungicides) were made in the sequence typical of the spray calendar for potatoes. A total of 15 treatments with the various compounds were made by spray application to the water surface at 0.2%, 1%, and 5% of the recommended label rates. Chemical fate and effects on ecosystem function and structure (phytoplankton, zooplankton, chlorophyll-a, macroinvertebrates, macrophytes, breakdown of plant litter) were investigated. To interpret the observed effects, treatment concentrations were also expressed in toxic units (TU), which describe the relative toxicity of the compounds with standard toxicity test organisms (Daphnia and algae). After treatment, each compound disappeared from the water phase within 2 d, with the exception of prosulfocarb, for which 50% dissipation time (DT50) values ranged between 6 and 7 d. At the 5% treatment level, an exposure peak of 0.9 TUalgae was observed, which resulted in short-term responses of pH, oxygen, and phytoplankton. At the 5% treatment level, exposure concentrations also exceeded 0.1 TUDaphnia, and this resulted in long-term effects on zooplankton and macroinvertebrates, some of which did not fully recover by the end of the present study. At the 1% treatment level, only slight transient effects were observed on a limited number of zooplankton and macro-invertebrate species and on pH. At the 0.2% level, no consistent treatment-related effects were observed. Most of the observed effects were consistent with the results from higher-tier and mesocosm studies with the individual compounds. Multi and repeated stress played a small role within the applied pesticide package, because of rapid dissipation of most substances and the absence of many simultaneous applications. This suggests that risk assessments based on the individual compounds would in this case have been sufficiently protective for their uses in a crop protection program.

The aim of the study was to compare the effects of the pyrethroid insecticide lambda-cyhalothrin (treated at 10, 25, 50, 100, 250 ng active ingredient a.i./L) on a drainage ditch ecosystem in spring and late summer. Microcosms (water volume approximately 430 L) were established using enclosures in a 50-cm-deep experimental ditch system containing communities typical of macrophyte-dominated freshwater ecosystems. Effects on macroinvertebrates, zooplankton, phytoplankton, macrophytes, and community metabolism were assessed and evaluated using univariate and multivariate statistical techniques. The macroinvertebrate community responded most clearly to treatment and, as anticipated, insects and crustaceans were among the most sensitive organisms. Statistical analysis showed that the underlying community structure was significantly different between the spring and summer experiments. However, the most sensitive species (Chaoborus obscuripes and Gammarus pulex) were abundant in spring as well as in late summer. In spring and late summer, only slight and transient effects were observed at the community level in the 10-ng/L treatment. Overall, the study did not show substantial differences in the responses of sensitive taxa between spring and late summer treatments, and effects thresholds were similar irrespective of season of treatment.

The fate and effects of the pyrethroid insecticide lambda-cyhalothrin were compared in mesotrophic (macrophyte-dominated) and eutrophic (phytoplankton-dominated) ditch microcosms (approximately 0.5 m3). Lambda-cyhalothrin was applied three times at one-week intervals at concentrations of 10, 25, 50, 100, and 250 ng/L. The rate of dissipation of lambda-cyhalothrin in the water column of the two types of test systems was similar. After 1 d, only 30% of the amount applied remained in the water phase. Initial, direct effects were observed primarily on arthropod taxa. The most sensitive species was the phantom midge (Chaoborus obscuripes). Threshold levels for slight and transient direct toxic effects were similar (10 ng/L) between types of test systems. At treatment levels of 25 ng/L and higher, apparent population and community responses occurred. At treatments of 100 and 250 ng/L, the rate of recovery of the macroinvertebrate community was lower in the macrophyte-dominated systems, primarily because of a prolonged decline of the amphipod Gammarus pulex. This species occurred at high densities only in the macrophyte-dominated enclosures. Indirect effects (e.g., increase of rotifers and microcrustaceans) were more pronounced in the plankton-dominated test systems, particularly at treatment levels of 25 ng/L and higher.

Results of previous laboratory studies suggest that high population density often buffers the effects of chemical stressors that predominately increase mortality. Mortality stressors act to release more resources for the survivors and, therefore, produce less-than-additive effects. By contrast, growth stressors are expected to have opposite results or more-than-additive effects. We investigated the effects of a growth inhibitor (lufenuron) on larval growth and survival of Chironomus riparius and examined its joint effects with density on population growth rate (PGR). Exposure to 60 microg/kg sediment or greater inhibited larval growth, and exposure to 88 microg/kg or greater often resulted in mortality before reaching emergence. The effects of lufenuron, however, differed with population density. At 88 microg/kg, mortalities and, to a lesser extent, reduced fecundity resulted in a reduction in PGR at low density. Conversely, when populations were initiated at high density, PGR was similar to that of controls, because the few survivors reached maturity sooner and started producing offspring earlier. The effect of density as a growth stressor therefore was stronger than the effect of lufenuron, which had effects similar to those of a mortality stressor and produced less-than-additive effects. Long-term studies under field conditions, however, are needed before less-than-additive effects are considered to be the norm.

Rapid progress in the field of genomics (the study of how an individual's entire genetic make-up, the genome, translates into biological functions) is beginning to provide tools that may assist our understanding of how chemicals can impact on human and ecosystem health. In many ways, if scientific and regulatory efforts in the 20th century have sought to establish which chemicals cause damage to ecosystems, then the challenge in ecotoxicology for the 21st century is to understand the mechanisms of toxicity to different wildlife species. In the human context, 'toxicogenomics' is the study of expression of genes important in adaptive responses to toxic exposures and a reflection of the toxic processes per se. Given the parallel implications for ecological (environmental) risk assessment, we propose the term 'ecotoxicogenomics' to describe the integration of genomics (transcriptomics, proteomics and metabolomics) into ecotoxicology. Ecotoxicogenomics is defined as the study of gene and protein expression in non-target organisms that is important in responses to environmental toxicant exposures. The potential of ecotoxicogenomic tools in ecological risk assessment seems great. Many of the standardized methods used to assess potential impact of chemicals on aquatic organisms rely on measuring whole-organism responses (e.g. mortality, growth, reproduction) of generally sensitive indicator species at maintained concentrations, and deriving 'endpoints' based on these phenomena (e.g. median lethal concentrations, no observed effect concentrations, etc.). Whilst such phenomenological approaches are useful for identifying chemicals of potential concern they provide little understanding of the mechanism of chemical toxicity. Without this understanding, it will be difficult to address some of the key challenges that currently face aquatic ecotoxicology, e.g. predicting toxicant responses across the very broad diversity of the phylogenetic groups present in aquatic ecosystems; estimating how changes at one ecological level or organisation will affect other levels (e.g. predicting population-level effects); predicting the influence of time-varying exposure on toxicant responses. Ecotoxicogenomic tools may provide us with a better mechanistic understanding of aquatic ecotoxicology. For ecotoxicogenomics to fulfil its potential, collaborative efforts are necessary through the parallel use of model microorganisms (e.g. Saccharomyces cerevisiae) together with aquatic (e.g. Danio rerio, Daphnia magna, Lemna minor and Xenopus tropicalis) and terrestrial (e.g. Arabidopsis thailiana, Caenorhabdites elegans and Eisenia foetida) plants, animals and microorganisms.

The toxicity of the pyrethroid insecticide lambda-cyhalothrin to freshwater invertebrates has been investigated using data from short-term laboratory toxicity tests and in situ bioassays and population-level effects in field microcosms. In laboratory tests, patterns of toxicity were consistent with previous data on pyrethroids. The midge Chaoborus obscuripes was most sensitive (48- and 96-h EC50 = 2.8 ng/L). Other insect larvae (Hemiptera, Ephemeroptera) and macrocrustacea (Amphipoda, Isopoda) were also relatively sensitive, with 48- and 96-h EC50 values between 10 and 100 ng/L. Generally, microcrustacea (Cladocera, Copepoda) and larvae of certain insect groups (Odonata and Chironomidae) were less sensitive, with 48-h EC50 values higher than 100 ng/L. Mollusca and Plathelminthes were insensitive and were unaffected at concentrations at and above the water solubility (5 microg/L). Generally, the EC50 values based on initial population responses in field enclosures were similar to values derived from laboratory tests with the same taxa. Also, the corresponding fifth and tenth percentile hazard concentrations (HC5 and HC10) were similar (laboratory HC5 = 2.7 ng/L and field HC5 = 4.1 ng/L; laboratory and field HC10 = 5.1 ng/L), at least when based on the same sensitive taxonomic groups (insects and crustaceans) and when a similar concentration range was taken into account. In the three field enclosure experiments and at a treatment level of 10 ng/L, consistent effects were observed for only one population (Chaoborus obscuripes), with recovery taking place within 3 to 6 weeks. The laboratory HC5 (2.7 ng/L) and HC10 (5.1 ng/L) based on acute EC50 values of all aquatic arthropod taxa were both lower than this 10 ng/L, a concentration that might represent the "regulatory acceptable concentration." The HC5 and HC10 values in this study in The Netherlands (based on static laboratory tests with freshwater arthropods) were very similar to those derived from a previous study in the United Kingdom (1.4 and 3.3 ng/L). This suggests that for pesticides like lambda-cyhalothrin, HC5 values based on static laboratory tests may provide a conservative estimate of the potential for community-level effects under field conditions. While these HC5 values are conservative for initial effects, they do not provide information on recovery potential, which may be important for regulatory decision-making.

Use of the insecticide lambda-cyhalothrin in agriculture may result in the contamination of water bodies, for example by spray drift. Therefore, the possible exposure of aquatic organisms to this insecticide needs to be evaluated. The exposure of the organisms may be reduced by the strong sorption of the insecticide to organic materials and its susceptibility to hydrolysis at the high pH values in the natural range. In experiments done in May and August, formulated lambda-cyhalothrin was mixed with the water body of enclosures in experimental ditches containing a bottom layer and macrophytes (at different densities) or phytoplankton. Concentrations of lambda-cyhalothrin in the water body and in the sediment layer, and contents in the plant compartment, were measured by gas-liquid chromatography at various times up to 1 week after application. Various water quality parameters were also measured. Concentrations of lambda-cyhalothrin decreased rapidly in the water column: 1 day after application, 24-40% of the dose remained in the water, and by 3 days it had declined to 1.8-6.5%. At the highest plant density, lambda-cyhalothrin residue in the plant compartment reached a maximum of 50% of the dose after 1 day; at intermediate and low plant densities, this maximum was only 3-11% of the dose (after 1-2 days). The percentage of the insecticide in the ditch sediment was 12% or less of the dose and tended to be lower at higher plant densities. Alkaline hydrolysis in the water near the surface of macrophytes and phytoplankton is considered to be the main dissipation process for lambda-cyhalothrin.

The partitioning, bioavailability, and toxicity of cypermethrin in water-sediment systems was investigated. Cypermethrin adsorbed extensively and rapidly, with an overall mean organic carbon (OC) adsorption partition coefficient (Koc) of 350,000, and approximately 99% adsorption occurred within 24 h. Bioavailability was measured via body burdens of Daphnia magna and Chironomus tentans. Mean biota-sediment accumulation factors (BSAFs), that is, the concentration in the organism as a proportion of the concentration in the sediment, decreased with increasing OC content. The BSAF values were 0.31, 0.14, and 0.08 for D. magna and 0.63, 0.19, and 0.08 for C. tentans, in 1, 3, and 13% OC sediments, respectively. The 10-d median lethal sediment concentrations (LC50s) of cypermethrin were 3.6, 18, and 32 mg/kg for Hyalella azteca and 13, 67, and 62 mg/kg for C. tentans in 1, 3, and 13% OC sediments, respectively. Predictions of aqueous concentrations at the LC50 in sediments (based on Koc) compared well to each other and to effect concentrations from studies in water alone, suggesting that equilibrium partitioning theory could be used reasonably to predict and normalize the toxicity of cypermethrin across sediments of differing OC content.

Aquatic exposure assessments for pesticides are generally based on laboratory studies performed in water alone or water sediment systems. Although aquatic macrophytes, which include a variety of bryophytes, macroalgae, and angiosperms, can be a significant component of many aquatic ecosystems, their impact on pesticide fate is generally not included in exposure assessments. To investigate the influence of aquatic plants on the fate and behavior of the pyrethroid insecticide lambda (lambda)-cyhalothrin, two laboratory experiments (to assess adsorption and degradation) and an indoor microcosm study (to assess fate under semirealistic conditions) were conducted. In the laboratory studies, adsorption to macrophytes was extensive and essentially irreversible, and degradation occurred rapidly by cleavage of the ester bond. In the indoor microcosm, which contained water, sediment, and macrophytes from a pond, degradation was also rapid, with DT50 and DT90 values of less than 3 and 19 h, respectively, for dissipation from the water column and of less than 3 and 56 h, respectively, for the whole system. For adsorptive and readily degraded pesticides like lambda-cyhalothrin, we conclude that macrophytes have considerable influence on fate and behavior in surface waters.

This is the first in a series of five papers that assess the risk of the cotton pyrethroids in aquatic ecosystems in a series of steps ranging from the analysis of effects data through modeling exposures in the landscape. Pyrethroid insecticides used on cotton have the potential to contaminate aquatic systems. The objectives of this study were to develop probabilistic estimates of toxicity distributions, to compare these among the pyrethroids, and to evaluate cypermethrin as a representative pyrethroid for the purposes of a class risk assessment of the pyrethroids. The distribution of cypermethrin acute toxicity data gave 10th centile values of 10 ng/L for all organisms, 6.4 ng/L for arthropods, and 380 ng/L for vertebrates. For bifenthrin, cyfluthrin, lambda-cyhalothrin, and deltamethrin, the 10th centile values for all organisms were 15, 12, 10, and 9 ng/L, respectively, indicating similar or somewhat lower toxicity than cypermethrin. For tralomethrin and fenpropathrin, the 10th centiles were <310 and 240 ng/L, respectively. The distribution of permethrin toxicity to all organisms, arthropods, and vertebrates gave 10th centiles of 180, 76, and 1600 ng/L, respectively, whereas those for fenvalerate were 37, 8, and 150 ng/L. With the exception of tralomethrin, the distributions of acute toxicity values had similar slopes, suggesting that the variation of sensitivity in a range of aquatic nontarget species is similar. The pyrethroids have different recommended field rates of application that are related to their efficacy, and the relationship between field rate and 10th centiles showed a trend. These results support the use of cypermethrin as a reasonable worst-case surrogate for the other pyrethroids for the purposes of risk assessment of pyrethroids as a class.

Estimates of potential aquatic exposure concentrations arising from the use of pyrethroid insecticides on cotton produced using conventional procedures outlined by the U.S. Environmental Protection Agency's Office of Pesticide Programs Environmental Fate and Effects Division seem unrealistically high. Accordingly, the assumptions inherent in the pesticide exposure assessment modeling scenarios were examined using remote sensing of a significant Mississippi, USA, cotton-producing county. Image processing techniques and a geographic information system were used to investigate the number and size of the water bodies in the county and their proximity to cotton. Variables critical to aquatic exposure modeling were measured for approximately 600 static water bodies in the study area. Quantitative information on the relative spatial orientation of cotton and water, regional soil texture and slope, and the detailed nature of the composition of physical buffers between agricultural fields and water bodies was also obtained. Results showed that remote sensing and geographic information systems can be used cost effectively to characterize the agricultural landscape and provide verifiable data to refine conservative model assumptions. For example, 68% of all ponds in the region have no cotton within 360 m and 92% of the ponds have no cotton within 60 m. Only 2% of ponds have cotton present in all directions around the ponds and within 120 m. These are significant modifications to conventional pesticide risk assessment exposure modeling assumptions and exemplify the importance of using landscape-level risk assessments to better describe the Mississippi cotton agricultural landscape. Incorporating spatially characterized landscape information into pesticide aquatic exposure scenarios is likely to have greater impact on the model output than many other refinements.

Since their introduction, synthetic pyrethroid insecticides have generated regulatory concerns regarding their toxicity to fish and aquatic invertebrates. In this paper we assess the potential for risks to aquatic ecosystems in cotton-growing areas, focusing on cypermethrin as a suitable representative of the pyrethroid class and static water bodies (ponds and lakes) as worst-case water bodies because of low levels of dilution. Reviews of cypermethrin effects under laboratory and field conditions have characterized the potential aquatic effects of the chemical. Also, a landscape-level exposure characterization has been conducted in a worst-case cotton-growing county, Yazoo County, Mississippi, USA, to provide a more realistic exposure characterization than is possible using standard model scenarios. Risks were characterized using the standard tier I and II approaches of the U.S. Environmental Protection Agency. In addition, a probabilistic risk assessment was conducted by comparing landscape-level exposure calculations for ponds and lakes in Yazoo County (modified tier II analysis) with distributions of laboratory effect concentrations and with data from field studies. Risk characterization using tier I and tier II models demonstrated a level of concern for certain aquatic organisms. However, modified tier II analysis showed that exposure concentrations are unlikely to exceed concentrations that might cause ecologically significant effects. Indeed, in the vast majority of cases, concentrations in the modified tier II analysis were several orders of magnitude lower than those at which effects would be predicted on the basis of laboratory and field data. The conclusion of minimal potential for adverse ecological effects was also supported by field studies, which showed that impacts on aquatic systems were negligible, even at concentrations many times higher than the modified tier II exposure concentrations.

Results of mesocosm and field studies with cypermethrin and esfenvalerate were analyzed and interpreted to support an ecological risk assessment of cotton pyrethroids in aquatic ecosystems. A core group of seven mesocosm studies conducted on two continents over the course of a decade were examined, and additional observations from mesocosm and field studies with these and other cotton pyrethroids were also brought to bear. The results for cypermethrin and esfenvalerate were remarkably consistent. They revealed a trend in sensitivity from amphipods, isopods, midges, mayflies, copepods, and cladocerans (most sensitive) to fish, snails, oligochaetes, and rotifers (least sensitive). With few exceptions, populations affected by pyrethroids in the mesocosms recovered to normal levels before the end of the year of exposure; most populations recovered within weeks. Factors presumed responsible for population recovery included internal refuges (areas of low exposure), resistant life stages, rapid generation times, and egg deposition by adults from outside the treated systems. Indirect effects on fish (which have been hypothesized to occur when invertebrate food sources are reduced) were not observed. The lowest-observed-adverse-effect concentrations for the overall ecosystems for cypermethrin and esfenvalerate corresponded to the 54th and 41st centiles of acute toxicity endpoints (LC50s) for arthropods measured in laboratory studies with these compounds, implying that a risk characterization based on 10th centiles would be highly conservative.

Environmental risk assessment is a key feature of regulations controlling the placing of new, and the maintenance of existing, chemicals products in the market place. For example, European Commission Directive 93/67/EC on Risk Assessment for New Notified Substances and Commission Regulation (EC) No. 1488/94 on Risk Assessment for Existing Substances requires that risk assessments be carried out for new and existing substances in the European Community. The process of environmental risk assessment seeks to determine the balance of probability of species and communities being damaged by chemical releases. The process relies upon a valid estimation of a predicted environmental concentration (PEC) in relevant environmental compartments and a predicted no effect concentration (PNEC) below which the organisms present in that compartment are unlikely to be significantly affected. If the PEC exceeds the PNEC there is a potential for damaging effects to occur. This article focuses on the determination of PNECs for risk assessment. Methods for determining a PNEC described in OECD Monograph 26 (1989, Report of the OECD Workshop on Ecological Effect Assessment, Paris, France, have been applied to data derived for the four chemicals lindane, 3,4-dichloroaniline, atrazine, and copper in a series of collaborative research projects funded by the European Commission., (Copyright 2000 Academic Press.)

This article presents a summary of a collaborative research program involving five European research groups, that was partly funded by the European Commission under its Environmental Research Program. The objective of the program was to develop aquatic toxicity tests that could be used to obtain data for inclusion at Level 2 of the Risk Evaluation Scheme for the Notification of Substances as required by the 7th Amendment to EC Directive 79/831/EEC. Currently only a very limited number of test methods have been described that can be used for this purpose and these are based on an even smaller number of test species. Tests based upon algae (Chlamydomonas reinhardi, Scenedesmus subspicatus, and Euglena gracilis), protozoa (Tetrahymena pyriformis), rotifera (Brachionus calyciflorus), crustacea (Gammarus pulex), and diptera (Chironomus riparius) were developed. The tests encompassed a range of end points and were evaluated against four reference chemicals: lindane, 3, 4-dichloroaniline (DCA), atrazine, and copper. The capacity of the tests to identify concentrations that are chronically toxic in the field was addressed by comparing the effects threshold concentrations determined in the laboratory tests with those determined for similar and/or related species and end points in stream and pond mesocosm studies. The lowest no-observed-effect concentrations (NOEC), EC(x), or LC(x) values obtained for lindane, atrazine, and copper were comparable with the lowest values obtained in the mesocosms. The lowest chronic NOEC determined for DCA using the laboratory tests was approximately 200 times higher than the lowest NOEC in the mesocosms., (Copyright 2000 Academic Press.)

Acute and sublethal toxicity of the organochlorine insecticide lindane to the amphipod crustacean Hyalella azteca was investigated. Acute experiments were conducted for a maximum test exposure period of 240 h with adult and neonate H. azteca. Median lethal concentrations (LC50s) determined for adult Hyalella included a 48-h LC50 of 47.6 &mgr;g/L and 240-h LC50 of 26.9 &mgr;g/L. For neonate H. azteca 24-, 48-, and 240-h LC50s were 29.5, 14.8, and 9.8 &mgr;g lindane/L, respectively. Neonate H. azteca were approximately three times more sensitive than adults. Two sublethal toxicity bioassays were developed based on the direct and indirect disruption of the precopulatory or mate guarding behavior of Hyalella. This reproductive behavior is readily quantifiable and of ecological significance as it is a vital component of the mating success of the species. The direct disruption bioassay examined the separation of precopulatory pairs maintained in control water and a range of lindane concentrations during a 24-h exposure period. Median separation times (ST50s) were determined and the LOEC was 24.4 &mgr;g lindane/L. The indirect disruption bioassay consisted of a test exposure period of just 4 h after which an invertebrate anesthetic solution was administered to induce separation of precopulatory pairs. The LOEC was 17.3 &mgr;g lindane/L, suggesting that the indirect precopulatory separation bioassay was comparable to the 24-h direct separation study. Both bioassays are rapid, relatively simple to perform, and have yielded effect concentrations that correspond with LC50 values determined using adult and neonate H. azteca life stages over more prolonged lindane exposures. Following some modification, these behavioral bioassays may be suitable for use in the hazard evaluation of sediments and for deployment as in situ toxicity tests.

: The fate and effects of two pyrethroid insecticides (lambda-cyhalothrin and cypermethrin) were investigated in replicated 25 m(3) pond mesocosms. Three pesticide treatments which simulated spray drift deposition were examined: 0.7 g a.i. ha(-1) cypermethrin and 0.17 and 1.7 g a.i. ha(-1) lambda-cyhalothrin. Based on the use rate and pesticidal activity of the chemicals, the cypermethrin and lower lambda-cyhalothrin rates were approximately equivalent. After applications, pyrethroid residues in the water column declined rapidly. Treatment-related effects were observed on some macroinvertebrate taxa, most notably the Asellidae and Gammaridae. Surfacedwelling insects also suffered initial knock-down, particularly in the 1.7 g a.i. ha(-1) lambda-cyhalothrin treatment, but there was recovery after the spray period. No adverse effects occurred on algae, macrophytes or zooplankton, but there were occasional enhancements (e.g. algal biomass and abundances of copepod nauplii and Rotifera) which may have been indirect effects. An overall comparison of the treatments indicated that the higher lambda-cyhalothrin rate had the greatest effects, whilst the cypermethrin application had a somewhat greater impact than the lower lambda-cyhalothrin treatment rate (due to effects on peracarid crustaceans). The study indicated that should spray drift occur at the levels expected for either pyrethroid's normal use patterns, potential impacts on natural aquatic ecosystems would be minor and transient.

The growth of the freshwater macroinvertebrates Gammarus pulex (L.) and Chironomus riparius Meigen exposed to 3,4-dichloroaniline in chambers within stream mesocosms was determined. DCA significantly affected the growth of neonate G. pulex and third instar C. riparius over 25 and 12 days, respectively. The no-observed-effect concentrations (NOECs) obtained in the tests were 0.08 mg DCA liter-1 (G. pulex) and 0.76 mg DCA liter-1 (C. riparius) and these are compared to toxicity data from other investigations. Inclusion of single-species bioassays in mesocosm studies provides complementary information on toxicant effects and indicates the suitability of the results of such tests (which may be routinely performed under laboratory conditions) for protecting particular ecosystems.

The acute and chronic toxicities of lindane to larvae of the freshwater insects Chironomus riparius Meigen, Chaoborus flavicans (Meigen), and Sigara striata (L.) were investigated in mesocosm compartments of an experimental pond. The following median lethal concentrations (LC50s) were determined: 240-hr LC50 of 2.0 micrograms lindane liter-1 for second instar C. riparius, 72-hr LC50 of 6.5 micrograms lindane liter-1 for fourth instar C. riparius, and 96-hr LC50s of 4.0 and 3.9 micrograms lindane liter-1 for fourth instar C. flavicans and fourth or fifth instar S. striata, respectively. Lindane significantly reduced the growth over 10 days of second instar C. riparius compared to that of the control at the treatment concentrations where larvae survived (1.0, 2.5, and 7.0 micrograms lindane liter-1). A significant increase in the median emergence time in comparison to that of the control was observed for C. riparius exposed to 0.8 and 2.0 micrograms lindane liter-1, with higher concentrations causing 100% mortality. The findings compare well with previously reported laboratory data on the toxicity of lindane to insects and support the methodology and results of a laboratory growth test for C. riparius.

The lethal toxicities of the four pollutants 3,4-dichloroaniline (DCA), atrazine, copper, and lindane were determined for the 2nd larval instar of the insect Chironomus riparius Meigen and the juvenile stage (2nd or 3rd moult) of the crustacean Gammarus pulex (L.). Median lethal concentrations (LC50s) were determined over a 240 h test period. The order of toxicity of the test chemicals is different for each species. For C. riparius, lindane was the most toxic, followed by copper, DCA, and atrazine. During the first 96 h of exposure, the order for G. pulex was copper, lindane, then DCA and atrazine with similar LC50 values. However, at 240 h lindane replaced copper as the most toxic chemical to G. pulex. The relative sensitivity of the two species was dependent on both the toxicant and the exposure period. The lethal concentrations determined for the four chemicals are compared to the results of other toxicity studies and discussed with respect to current standard test methods.