Human risk associated with exposure to mixtures of antiandrogenic chemicals evaluated using in vitro hazard and human biomonitoring data.

BACKGROUND: Scientific evidence for underestimated toxicity from unintentional exposure to chemical mixtures is mounting. Yet, harmonized approaches on how to assess the actual risk of mixtures is lacking. As part of the European Joint programme 'Human Biomonitoring for Europe' we explored a novel methodology for mixture risk assessment of chemicals affecting male reproductive function. METHODOLOGY: We explored a methodology for chemical mixture risk assessment based on human in vitro data combined with human exposure data, thereby circumventing the drawbacks of using hazard data from rodents and estimated exposure intake levels. Human androgen receptor (hAR) antagonism was selected as the most important molecular initiating event linked to adverse outcomes on male reproductive health. RESULTS: Our work identified 231 chemicals able to interfere with hAR activity. Among these were 61 finally identified as having both reliable hAR antagonist and human biomonitoring data. Calculation of risk quotients indicated that PCBs (118, 138, 157), phthalates (BBP, DBP, DIBP), benzophenone-3, PFOS, methylparaben, triclosan, some pesticides (i.e cypermethrin, ß-endosulfan, methylparathion, p,p-DDE), and a PAH metabolite (1-hydroxypyrene) contributed to the mixture effect. The major chemical mixture drivers were PCB 118, BBP, PFOS, DBP, and the UV filter benzophenone-3, together contributing with 75% of the total mixture effect that was primarily driven by high exposure values. CONCLUSIONS: This viable way forward for mixture risk assessment of chemicals has the advantages of (1) being a more comprehensive mixture risk assessment also covering data-poor chemicals, and (2) including human data only. However, the approach is subjected to uncertainties in terms of in vitro to in vivo extrapolation, it is not ready for decision making, and needs further development. Still, the results indicate a concern for adverse effects on reproductive function in highly exposed boys, especially when considering additional exposure to data-poor chemicals and chemicals acting by other mechanisms of action.

The German Environmental Survey for Children and Adolescents 2014-2017 (GerES V) - Study population, response rates and representativeness.

The German Environmental Survey (GerES) is a population-representative, cross-sectional study on environmental exposures of the general population of Germany. GerES has repeatedly been conducted since 1985 by the German Environment Agency (UBA) in close collaboration with the Health Interview and Examination Surveys of the Robert Koch Institute (RKI). In the German Environmental Survey for Children and adolescents 2014-2017 (GerES V) pollutants and other environmental stressors were measured in human samples as well as in the homes of 3- to 17-year-old children and adolescents. Interviews were conducted about health-related behaviors and living conditions. The GerES V basic program encompassed examinations of whole blood, blood plasma, morning urine and drinking water samples, measurements of ultrafine particles and noise levels, comprehensive standardized interviews, and self-administrated questionnaires. Additional modules on volatile organic compounds and aldehydes, particulate matter (PM2.5) in indoor air, organic compounds in drinking water and pollutants in house dust were conducted in subsamples. Potential GerES V participants were identified and attained by the RKI from those participants who were examined and interviewed for the cross-sectional component of the second follow-up to the German Health Interview and Examination Survey for Children and Adolescents (KiGGS Wave 2). The gross sample of GerES V comprised 3031 children and adolescents of which 2294 finally took part in the survey. This equals a total response rate of 75.7 %. Response rates varied, depending on region, type of municipality, age and sex, from 66.0 % to 78.3 %. By calculating individual case weights, discrepancies due to sample design and non-response between the GerES V sample and the whole population could be considered in statistical analysis. Therefore, the representativeness of the GerES V results with regard to age, sex, community size and region was assured.

Disentangling multiple chemical and non-chemical stressors in a lotic ecosystem using a longitudinal approach.

Meeting ecological and water quality standards in lotic ecosystems is often failed due to multiple stressors. However, disentangling stressor effects and identifying relevant stressor-effect-relationships in complex environmental settings remain major challenges. By combining state-of-the-art methods from ecotoxicology and aquatic ecosystem analysis, we aimed here to disentangle the effects of multiple chemical and non-chemical stressors along a longitudinal land use gradient in a third-order river in Germany. We distinguished and evaluated four dominant stressor categories along this gradient: (1) Hydromorphological alterations: Flow diversity and substrate diversity correlated with the EU-Water Framework Directive based indicators for the quality element macroinvertebrates, which deteriorated at the transition from near-natural reference sites to urban sites. (2) Elevated nutrient levels and eutrophication: Low to moderate nutrient concentrations together with complete canopy cover at the reference sites correlated with low densities of benthic algae (biofilms). We found no more systematic relation of algal density with nutrient concentrations at the downstream sites, suggesting that limiting concentrations are exceeded already at moderate nutrient concentrations and reduced shading by riparian vegetation. (3) Elevated organic matter levels: Wastewater treatment plants (WWTP) and stormwater drainage systems were the primary sources of bioavailable dissolved organic carbon. Consequently, planktonic bacterial production and especially extracellular enzyme activity increased downstream of those effluents showing local peaks. (4) Micropollutants and toxicity-related stress: WWTPs were the predominant source of toxic stress, resulting in a rapid increase of the toxicity for invertebrates and algae with only one order of magnitude below the acute toxic levels. This toxicity correlates negatively with the contribution of invertebrate species being sensitive towards pesticides (SPEARpesticides index), probably contributing to the loss of biodiversity recorded in response to WWTP effluents. Our longitudinal approach highlights the potential of coordinated community efforts in supplementing established monitoring methods to tackle the complex phenomenon of multiple stress.

Long-term effects of a catastrophic insecticide spill on stream invertebrates.

Accidental spills or illegal discharges of pesticides in aquatic ecosystems can lead to exposure levels that strongly exceed authorized pesticide concentrations, causing major impacts on aquatic ecosystems. Such short-term events often remain undetected in regular monitoring programs with infrequent sampling. In early spring 2015, we identified a catastrophic pesticide spill with the insecticide cypermethrin in the Holtemme River, Germany. Based on existing pre-event macroinvertebrate community data, we monitored the effects and recovery of the macroinvertebrate community for more than two years after the spill. Strong short-term effects were apparent for all taxa with the exception of Chironomidae and Tubificidae. Effects could also be observed on the community level as total abundance, taxa number and biomass strongly decreased. Total abundance and taxa number showed a fast recovery. Regarding long-term effects, the total biomass remained substantially below the pre-contamination level (76%) until the end of the study. Also the abundances of three taxa (Gammarus, Leuctra, Limnius Ad.) did not return to levels prior to the spill even after 26 months. This lack of the taxon-specific recovery was likely due to their long generation time and a low migration ability due to a restricted connectivity between the contaminated site and uncontaminated stream sections. These factors proved to be stronger predictors for the recovery than the pesticide tolerance. We revealed that the biological indicators SPEARpesticides and share of Ephemeroptera, Plecoptera and Trichoptera (EPT) are not suitable for the identification of such extreme events, when nearly all taxa are eradicated. Both indicators are functioning only when repeated stressors initiate long-term competitive replacement of sensitive by insensitive taxa. We conclude that pesticide spills can have significant long-term effects on stream macroinvertebrate communities. Regular ecological monitoring is imperative to identify such ecosystem impairments, combined with analytical chemistry methods to identify the potential sources of spills.

Learning from previous work and finding synergies in the domains of public and environmental health: EU-funded projects BRIDGE Health and HBM4EU.

BACKGROUND: During the last decade, the European Union initiated several projects in the domains of public and environmental health. Within this framework, BRIDGE Health (Bridging Information and Data Generation for Evidence-based Health policy and Research) and HBM4EU (European human biomonitoring initiative) have been implemented. Whereas, the focus of BRIDGE Health was towards a sustainable and integrated health information system (HIS), the aim of HBM4EU is to improve evidence of the internal exposure of European citizens to environmental chemicals by human biomonitoring (HBM) and the impact of internal exposure on health. As both, environmental and public health determinants are important for health promotion, disease prevention and policy, BRIDGE Health and HBM4EU have overlapping aims and outcomes. In order to improve health information regarding public health and environmental health issues, best use and exchange of respective networks and project results is necessary. METHODS: Both projects have implemented health information (HI) and HBM tasks in order to provide adequate environmental and public health information of the European population. Synergies of the projects were identified in the working progress and because of overlapping networks and experts a focused analysis of both projects was envisaged. This paper elaborates on the aims and outcomes of both projects and the benefit of merging and channelling research results for the use of better health information and policy making that may be of relevance for any other project in these research fields. RESULTS: The need for focused exchanges and collaborations between the projects were identified and benefits of exchanges were highlighted for the specific areas of indicator development, linkage of data repositories and the combination of HBM studies and health examination surveys (HES). Further recommendations for a European wide harmonisation among different tasks in the fields of public health and environmental health are being developed. CONCLUSIONS: Lessons learned from HBM4EU and BRIDGE Health show that continuous efforts must be undertaken, also by succeeding projects, to guarantee the exchange between public health and environmental health issues. Networks covering both are essential to provide better evidence of knowledge. The experiences from BRIDGE Health and HBM4EU give a valuable input for any future activity in these domains. Avoiding overlaps and streamlining further exchange of public health and environmental health contributes to best use of research results and allows to develop new strategies and tools for improvement of health information and thus enhances people's health and well-being.

Species occurrence relates to pesticide gradient in streams.

Freshwater communities are threatened worldwide, with pesticides being one of the main stressors for vulnerable invertebrates. Whereas the effects of pesticides on communities can be quantified by trait-based bioindicators such as SPEARpesticides, single species' responses remain largely unknown. We used the bioindicator SPEARpesticides to predict the toxic pressure from pesticides in 6942 macroinvertebrate samples from 4147 sites during the period 2004 to 2013, obtained by environmental authorities in Germany, and classified all samples according to their magnitude of pesticide pressure. Along this gradient of pesticide pressure, we quantified the occurrence of 139 macroinvertebrate species. We identified 71 species characterized by decreasing occurrence with increasing pesticide pressure. These 'decreasing species', mainly insects, occurred at a frequency of 19.7% at sites with reference conditions and decreased to 1.7% at sites with the highest pesticide pressure. We further determined 55 'nonspecific species' with no strong response as well as 13 'increasing species', mainly Gastropoda, Oligochaeta and Diptera, which showed an increase of frequency from 1.8% at sites with reference conditions to 11.4% at sites with the highest pesticide pressure. Based on the change in frequency we determined the pesticide vulnerability of single species, expressed as Pesticide Associated Response (PARe). Furthermore, a trait analysis revealed that species' occurrence may additionally depend on oxygen demand and, to a lesser extent on substrate preference, whereas no significant effect of feeding and respiration type could be found. Our results provide the first extensive pesticide vulnerability ranking for single macroinvertebrate species based on empirical large-scale field data.

Statement on advancing the assessment of chemical mixtures and their risks for human health and the environment.

The number of anthropogenic chemicals, manufactured, by-products, metabolites and abiotically formed transformation products, counts to hundreds of thousands, at present. Thus, humans and wildlife are exposed to complex mixtures, never one chemical at a time and rarely with only one dominating effect. Hence there is an urgent need to develop strategies on how exposure to multiple hazardous chemicals and the combination of their effects can be assessed. A workshop, "Advancing the Assessment of Chemical Mixtures and their Risks for Human Health and the Environment" was organized in May 2018 together with Joint Research Center in Ispra, EU-funded research projects and Commission Services and relevant EU agencies. This forum for researchers and policy-makers was created to discuss and identify gaps in risk assessment and governance of chemical mixtures as well as to discuss state of the art science and future research needs. Based on the presentations and discussions at this workshop we want to bring forward the following Key Messages.

Realistic pesticide exposure through water and food amplifies long-term effects in a Limnephilid caddisfly.

Pesticides are increasingly recognized as relevant stressors in stream ecosystems. Stream biota is exposed to pesticides with low water solubility, e.g. pyrethroid insecticides, via water, habitat, and food. However, long-term effects of simultaneous exposure pathways are unknown. In this context, we conducted a microcosm experiment with the caddisfly Anabolia nervosa exposing the larvae to the pyrethroid insecticide esfenvalerate (EFV) at 0.1 and 1.0µgL-1 via (i) water, (ii) food or a (iii) combination of water and food. Combined exposure through water and food significantly reduced emergence by 60% and significantly postponed emergence timing at the highest EFV level, whereas none of the single-phase exposures showed significant effects. Moreover, our study revealed that successfully emerged females from the highest biphasic treatment level were characterised by altered composition of storage lipids indicative of reduced energy reserves. Consequently, a realistic test scenario that represents simultaneous exposure of organisms and their food may reveal substantially increased long term effects of pyrethroids when compared with current ecological risk assessment applying only single phase exposure. We recommend that relevant concurrent exposure routes of pesticides should be considered in order to derive realistic regulatory acceptable concentrations of pesticides.