Linking chemical surface water monitoring and pesticide regulation in selected European countries.

The progress in chemical analytics and understanding of pesticide dynamics in surface waters allows establishing robust data on compounds with frequent exceedances of quality standards. The current chemical, temporal, and spatial coverage of the pesticide monitoring campaigns differs strongly between European countries. A questionnaire revealed differences in monitoring strategies in seven selected European countries; Nordic countries prioritize temporal coverage, while others focus on spatial coverage. Chemical coverage has increased, especially for non-polar classes like synthetic pyrethroids. Sweden combines monitoring data with agricultural practices for derived quantities, while the Netherlands emphasizes spatial coverage to trace contamination sources. None of the EU member states currently has established a process for linking chemical surface water monitoring data with regulatory risk assessment, while Switzerland has recently established a legally defined feedback loop. Due to their design and objectives, most strategies do not capture concentration peaks, especially 2-week composite samples, but also grab samples. Nevertheless, for substances that appear problematic in many data sets, the need for action is evident even without harmonization of monitoring programs. Harmonization would be beneficial, however, for cross-national assessment including risk reduction measures.

Analysis and dissipation of the antiparasitic agent ivermectin in cattle dung under different field conditions.

Cattle treated with the veterinary parasiticide ivermectin fecally excrete residues. The authors report the exposition and dissipation characteristics of these residues in dung of ivermectin-treated cattle and in soil beneath this dung on pastures in Canada, France, Switzerland, and The Netherlands. Residues were quantified for dung collected from cattle after 3 d, 7 d, 14 d, and 28 d posttreatment and subsequently exposed in the field for up to 13 mo. The authors optimized a high-performance liquid chromatography-fluorescence detection method to detect ivermectin residues in dung and soil matrices. They showed that a solid phase extraction and purification step generally can be eliminated to reduce the time and cost of these analyses. They also found that the addition of water to relatively dry samples improves the extraction efficiency of residues. They then analyzed the field samples to document differences in ivermectin dissipation in cattle dung among sites, with 50% dissipation times of up to 32 d and 90% dissipation times >396 d. They further showed that the dissipation characteristics of residues are comparable between dung of ivermectin-treated cattle and dung to which ivermectin has been added directly. Lastly, they report the first use of a desorption electrospray ionization-high-resolution-mass spectrometric method to detect residues of metabolites in a dung matrix. Environ Toxicol Chem 2016;35:1924-1933. © 2016 SETAC.

Nontarget effects of ivermectin residues on earthworms and springtails dwelling beneath dung of treated cattle in four countries.

The authorization of veterinary medicinal products requires that they be assessed for nontarget effects in the environment. Numerous field studies have assessed these effects on dung organisms. However, few studies have examined effects on soil-dwelling organisms, which might be exposed to veterinary medicinal product residues released during dung degradation. The authors compared the abundance of earthworms and springtails in soil beneath dung from untreated cattle and from cattle treated 0 d, 3 d, 7 d, 14 d, and 28 d previously with ivermectin. Study sites were located in different ecoregions in Switzerland (Continental), The Netherlands (Atlantic), France (Mediterranean), and Canada (Northern Mixed Grassland). Samples were collected using standard methods from 1 mo to 12 mo after pat deposition. Ivermectin concentrations in soil beneath dung pats ranged from 0.02 mg/kg dry weight (3 mo) to typically <0.006 mg/kg dry weight (5-7 mo). Earthworms were abundant and species-rich at the Swiss and Dutch sites, less common with fewer species at the French site, and essentially absent at the Canadian site. Diverse but highly variable communities of springtails were present at all sites. Overall, results showed little effect of residues on either earthworms or springtails. The authors recommend that inclusion of soil organisms in field studies to assess the nontarget effects of veterinary medicinal products be required only if earthworms or springtails exhibit sensitivity to the product in laboratory tests. Environ Toxicol Chem 2016;35:1959-1969. © 2015 SETAC.

A four-country ring test of nontarget effects of ivermectin residues on the function of coprophilous communities of arthropods in breaking down livestock dung.

By degrading the dung of livestock that graze on pastures, coprophilous arthropods accelerate the cycling of nutrients to maintain pasture quality. Many veterinary medicinal products, such as ivermectin, are excreted unchanged in the dung of treated livestock. These residues can be insecticidal and may reduce the function (i.e., dung-degradation) of the coprophilous community. In the present study, we used a standard method to monitor the degradation of dung from cattle treated with ivermectin. The present study was performed during a 1-yr period on pastures in Canada, France, The Netherlands, and Switzerland. Large effects of residue were detected on the coprophilous community, but degradation of dung was not significantly hampered. The results emphasize that failure to detect an effect of veterinary medicinal product residues on dung-degradation does not mean that the residues do not affect the coprophilous community. Rather, insect activity is only one of many factors that affect degradation, and these other factors may mask the nontarget effect of residues. Environ Toxicol Chem 2016;35:1953-1958. © 2015 SETAC.

Validation of a standard field test method in four countries to assess the toxicity of residues in dung of cattle treated with veterinary medical products.

Registration of veterinary medical products includes the provision that field tests may be required to assess potential nontarget effects associated with the excretion of product residues in dung of treated livestock (phase II, tier B testing). However, regulatory agencies provide no guidance on the format of these tests. In the present study, the authors report on the development of a standardized field test method designed to serve as a tier B test. Dung was collected from cattle before and up to 2 mo after treatment with a topical application of a test compound (ivermectin). Pats formed of dung from the different treatments were placed concurrently in the field to be colonized by insects. The abundance, richness, and diversity of insects developing from egg to adult in these pats were compared across treatments using analysis of variance tests. Regression analyses were used to regress abundance, richness, and diversity against residue concentrations in each treatment. Results of the regression were used to estimate mean lethal concentration (LC50) values. The robustness of the method and the repeatability of its findings were assessed concurrently in 4 countries (Canada, France, Switzerland, and The Netherlands) in climatically diverse ecoregions. Results were generally consistent across countries, and support the method's formal adoption by the European Union to assess the effects of veterinary medical product residues on the composition and diversity of insects in dung of treated livestock. Environ Toxicol Chem 2016;35:1934-1946. © 2015 Crown in the right of Canada. Published by Wiley Periodicals Inc., on behalf of SETAC.

Towards a renewed research agenda in ecotoxicology.

New concerns about biodiversity, ecosystem services and human health triggered several new regulations increasing the need for sound ecotoxicological risk assessment. The PEER network aims to share its view on the research issues that this challenges. PEER scientists call for an improved biologically relevant exposure assessment. They promote comprehensive effect assessment at several biological levels. Biological traits should be used for Environmental risk assessment (ERA) as promising tools to better understand relationships between structure and functioning of ecosystems. The use of modern high throughput methods could also enhance the amount of data for a better risk assessment. Improved models coping with multiple stressors or biological levels are necessary to answer for a more scientifically based risk assessment. Those methods must be embedded within life cycle analysis or economical models for efficient regulations. Joint research programmes involving humanities with ecological sciences should be developed for a sound risk management.

How to test nontarget effects of veterinary pharmaceutical residues in livestock dung in the field.

To register veterinary medicinal products (VMPs) as parasiticides on pastured animals, legislation in the European Union requires an environmental risk assessment to test the potential nontarget effects of fecal residues on dung-dwelling organisms. Products with adverse effects in single-species laboratory tests require further, higher-tier testing to assess the extent of these effects on entire communities of dung-dwelling organisms under more realistic field or semifield conditions. Currently, there are no documents specifically written to assist researchers in conducting higher-tier tests or to assist regulators in interpreting the results of such tests in an appropriate context. Here we provide such a document, written by members of the SETAC Advisory Group DOTTS (Dung Organism Toxicity Testing Standardization) with research experience on dung fauna in central and southern Europe, Canada, Australia, and South Africa. This document briefly reviews the organisms that make up the dung community and their role in dung degradation, identifies key considerations in the design and interpretation of experimental studies, and makes recommendations on how to proceed.

Ecological vulnerability in wildlife: application of a species-ranking method to food chains and habitats.

Nature development in The Netherlands is often planned on contaminated soils or sediments. This contamination may present a risk for wildlife species desired at those nature development sites and must be assessed by specific risk assessment methods. In a previous study, we developed a method to predict ecological vulnerability in wildlife species by using autecological data and expert judgment; in the current study, this method is further extended to assess ecological vulnerability of food chains and terrestrial and aquatic habitats typical for The Netherlands. The method is applied to six chemicals: Cd, Cu, Zn, dichlorodiphenyltrichloroethane, chlorpyrifos, and ivermectin. The results indicate that species in different food chains differ in vulnerability, with earthworm-based food chains the most vulnerable. Within and between food chains, vulnerability varied with habitat, particularly at low trophic levels. The concept of habitat vulnerability was applied to a case study of four different habitat types in floodplains contaminated with cadmium and zinc along the river Dommel, The Netherlands. The alder floodplain forest habitat contained the most vulnerable species. The differences among habitats were significant for Cd. We further conclude that the method has good potential for application in mapping of habitat vulnerability.

Wildlife vulnerability and risk maps for combined pollutants.

Ecological risk and vulnerability maps can be used to improve the analysis of pollutant risks and communication to stakeholders. Often, such maps are made for one pollutant at the time. We used the results of wildlife vulnerability analysis, a novel trait-based risk assessment approach, to map overall vulnerability of habitats in Denmark to various metals and one insecticide. These maps were combined with maps of estimated soil concentrations for the same compounds divided by their Maximum Permissible Concentrations. This combination yielded relative risk maps that can be used to assess where the highest risk conditions to wildlife from these individual pollutants in Denmark occur (hot spot identification). In order to show how cumulative risk maps can be made, the maps of the individual pollutants were combined assuming different mechanisms of joint toxicity: no addition, concentration addition, antagonism and synergism. The study demonstrated that with an accurate set of geographical and ecological data one can use the results of vulnerability analysis to make relevant ecological risk maps that show hot spot areas for risks of single or cumulative risks from soil pollutants.

Environmental risk mapping of pollutants: state of the art and communication aspects.

Risk maps help risk analysts and scientists to explore the spatial nature of the effects of environmental stressors such as pollutants. The development of Geographic Information Systems over the past few decades has greatly improved spatial representation and analysis of environmental information and data. Maps also constitute a powerful tool to communicate the outcome of complex environmental risk assessment to stakeholders such as the general public and policy makers. With appropriate cartography one can improve communication and thus bridge the gap between experts and users. Appropriate risk communication is pivotal to risk management, decision making and implementation and may prevent unnecessary concern about environmental pollutants. However, at present few risk maps are specifically tailored to meet the demands of such defined uses. This paper presents an overview of the most important types of risk maps that can be distinguished using examples from the scientific literature: contamination maps, exposure maps, hazard maps, vulnerability maps and 'true' risk maps. It also discusses, in a general way, the most important issues that need to be addressed when making risk maps for communication purposes: risk perception, target audience, scale and spatial aggregation and visualisation such as use of colours and symbols. Finally, some general rules of thumb are given for making environmental risk maps for communication purposes.

Ecological vulnerability in wildlife: an expert judgment and multicriteria analysis tool using ecological traits to assess relative impact of pollutants.

Nature development in The Netherlands often is planned on contaminated soils and sediments of former agricultural land and in floodplain areas; however, this contamination may present a risk to wildlife species desired at those nature development sites. Specific risk assessment methods are needed, because toxicological information is lacking for most wildlife species. The vulnerability of a species is a combination of its potential exposure, sensitivity to the type of pollutant, and recovery capacity. We developed a new method to predict ecological vulnerability in wildlife using autecological information. The analysis results in an ordinal ranking of vulnerable species. The method was applied to six representative contaminants: copper and zinc (essential metals, low to medium toxicity), cadmium (nonessential metal, high toxicity), DDT (persistent organic pesticide, high toxicity), chlorpyrifos (persistent organophosphate insecticide, high toxicity), and ivermectin (persistent veterinary pharmaceutical, low to medium toxicity). High vulnerability to the essential metals copper and zinc was correlated with soil and sediment habitat preference of a species and with r-strategy (opportunistic strategy suited for unstable environments). Increased vulnerability to the bioaccumulating substances cadmium and DDT was correlated with higher position of a species in the food web and with life span and K-strategy (equilibrium strategy suited for stable environments). Vulnerability to chlorpyrifos and ivermectin was high for species with a preference for soil habitats. The ecological vulnerability analysis has potential to further our abilities in risk assessment.

An integrated assessment of estrogenic contamination and biological effects in the aquatic environment of The Netherlands.

An extensive study was carried out in the Netherlands on the occurrence of a number of estrogenic compounds in surface water, sediment, biota, wastewater, rainwater and on the associated effects in fish. Compounds investigated included natural and synthetic hormones, phthalates, alkylphenol(ethoxylate)s and bisphenol-A. The results showed that almost all selected (xeno-)estrogens were present at low concentrations in the aquatic environment. Locally, they were found at higher levels. Hormones and nonylphenol(ethoxylate)s were present in concentrations that are reportedly high enough to cause estrogenic effects in fish. Field surveys did not disclose significant estrogenic effects in male flounder (Platichthys flesus) in the open sea and in Dutch estuaries. Minor to moderate estrogenic effects were observed in bream (Abramis brama) in major inland surface waters such as lowland rivers and a harbor area. The prevalence of feminizing effects in male fish is largest in small regional surface waters that are strongly influenced by sources of potential hormone-disrupting compounds. High concentrations of plasma vitellogenin and an increased prevalence of ovotestes occurred in wild male bream in a small river receiving a considerable load of effluent from a large sewage treatment plant. After employing in vitro and in vivo bioassays, both in situ and in the laboratory, we conclude that in this case hormones (especially 17 alpha-ethynylestradiol) and possibly also nonylphenol(ethoxylate)s are primarily responsible for these effects.

Responses in sediment bioassays used in the Netherlands: can observed toxicity be explained by routinely monitored priority pollutants?

In order to identify the cause of toxicity in sediments and suspended matter, a large number of samples with different degrees of contamination was taken at various locations in The Netherlands. Standard acute bioassays were carried out with the bacterium Vibrio fischeri, the rotifer Brachionus calyciflorus and the anostracan Thamnocephalus platyurus. Chronic standard tests were performed using the water flea Daphnia magna and larvae of the midge Chironomus riparius. Some novel bioassays were performed as well. Most toxic effects observed in standard bioassays with sediments from polluted sediments (class 3 and 4 on a scale of 0-4 according to the Dutch criteria) could be partly explained by toxic concentrations of known persistent priority pollutants, mainly heavy metals and occasionally polycyclic aromatic hydrocarbons. In some of the samples, ammonia toxicity was a confounding factor during testing. Suspended matter from the Meuse river at Eijsden, which may be considered as 'new' sediment (pollution class 2), was moderately to highly toxic in almost all bioassays. This could have been associated with a combination of heavy metals, PAHs and ammonia. At two locations from the Lake IJssel area with no apparent persistent pollution, moderate and strong effects were nonetheless observed in invertebrate tests. This might have been due to agricultural run-off of pesticides, which are not routinely measured in sediments. A few effects on V. fischeri in canals and a small stream could not be explained with standard chemical analysis, but seemed associated with the outlets of sewage water treatment plants and industrial effluents. Additional chemical analysis of pore water samples from five selected sediments yielded more identified substances such as phtalates, decanes, cosanes and fragrances, but it was estimated that their contribution to the effects observed on V. fischeri, D. magna and C. riparius was negligible.

Estrogenic effects in fish in The Netherlands: some preliminary results.

Recently, a large-scale field study in The Netherlands has focused on the effects of estrogenic contaminants on feral fish populations. The freshwater bream (Abramis brama) and the estuarine flounder (Platichthys flesus) were sampled at a large number of locations in the spring and autumn of 1999. Concentrations of the yolk protein vitellogenin (VTG) in blood plasma of male flounders were small at most sites. At two sites, however, moderately elevated concentrations were found in autumn. Both sites were situated in the same industrial harbour zone also receiving effluent from sewage treatment works. At many sites VTG levels in male bream were significantly greater than at the control site. The greatest concentrations were observed in individuals collected from a small stream, close to the discharge of a relatively large municipal waste water treatment plant. This was also the only site where considerable intersex occurred; 37% of male bream exhibited ovotestes. Ovotestis was not observed in any of the male flounder captured. The results from The Netherlands are briefly discussed and compared with the well-known case studies in the UK.

Biological measurement of estrogenic activity in urine and bile conjugates with the in vitro ER-CALUX reporter gene assay.

Although estrogens are excreted as biologically inactive conjugates, they can be reconverted to an active form, possibly by bacteria. A simple method was developed to deconjugate estrogen metabolites present in human urine and fish bile back to active estrogens by enzymatic hydrolysis with beta-glucuronidase or live Escherichia coli cells. Deconjugated extracts were tested for estrogenic activity in the in vitro stable estrogen receptor-mediated chemical-activated luciferase gene expression (ER-CALUX) assay. Estrogen glucuronides in urine obtained from human males and females were effectively converted to active forms after incubation with beta-glucuronidase or E. coli. The highest estrogenic activity was found in deconjugated metabolites from urine of a pregnant woman, in which levels up to 3,000 nmol estradiol equivalents per liter of urine were found after overnight incubation of urine with E. coli. Bile sampled from male bream and flounder from various freshwater and marine locations was also deconjugated and a good correlation was found between high biliary estrogenic activity and elevated levels of xenoestrogenic activity in surface water as well as in plasma vitellogenin. Therefore, the measurement of deconjugated bile could form a useful (indirect) biomarker for internal dose of xenoestrogens in male fish.