Water Framework Directive micropollutant monitoring mirrors catchment land use: Importance of agricultural and urban sources revealed.

River monitoring programs worldwide consistently unveil micropollutant concentrations (pesticide, pharmaceuticals, and industrial chemicals) exceeding regulatory quality targets with deteriorating effects on aquatic communities. However, both the composition and individual concentrations of micropollutants are likely to vary with the catchment land use, in particular regarding urban and agricultural area as the primary sources of micropollutants. In this study, we used a dataset of 109 governmental monitoring sites with micropollutants monitored across the Federal State of North Rhine-Westphalia, Germany, to investigate the relationship between high-resolution catchment land use (distinguishing urban, forested and grassland area as well as 22 different agricultural crop types) and 39 micropollutants using Linear Mixed Models (LMMs). Ecotoxicological risks were indicated for mixtures of pharmaceutical and industrial chemicals for 100 % and for pesticides for 55 % of the sites. The proportion of urban area in the catchment was positively related with concentrations of most pharmaceuticals and industrial chemicals (R2 up to 0.54), whereas the proportions of grassland and forested areas generally showed negative relations. Cropland overall showed weak positive relationships with micropollutant concentrations (R2 up to 0.29). Individual crop types, particularly vegetables and permanent crops, showed higher relations (R2 up to 0.46). The findings suggest that crop type-specific pesticide applications are mirrored in the detected micropollutant concentrations. This highlights the need for high-resolution spatial land use to investigate the magnitude and dynamics of micropollutant exposure and relevant pollution sources, which would remain undetected with highly aggregated land use classifications. Moreover, the findings imply the need for tailored management measures to reduce micropollutant concentrations from different sources and their related ecological effects. Urban point sources, could be managed by advanced wastewater treatment. The reduction of diffuse pollution from agricultural land uses requires additional measures, to prevent pesticides from entering the environment and exceeding regulatory quality targets.

Linking wastewater treatment plant effluents to water quality and hydrology: Effects of multiple stressors on fish communities.

Wastewater treatment plants (WWTPs) are essential for maintaining a good water quality of surface waters. However, WWTPs are also associated with water quality deterioration and hydro-morphological alteration. Riverine communities respond to these stressors with changes in their community structure, abundance and diversity. In this study, we used a dataset of 94 monitoring sites across North Rhine-Westphalia, Germany to investigate the influence of WWTPs on the water quality and hydro-morphological quality in river sections downstream of WWTP effluents. More specifically, we analyzed the effects of the percentage of WWTP effluents (in relation to median base flow) on four stressor groups (physico-chemistry, micropollutants, hydrological and morphological alteration) using Linear Mixed Models (LMM). Furthermore, we assessed the impact of a selection of twelve ecologically relevant stressor variables reflecting water quality deterioration and hydro-morphological alteration on reference fish communities using Canonical Correspondence Analysis (CCA). The percentage of WWTP effluents was correlated with water quality, especially with toxic units of a wide range of pharmaceuticals including diclofenac, venlafaxine and sulfamethoxazole (R² up to 0.54) as well as specific pesticides (e.g., terbutryn: R² = 0.33). The correlation of percent WWTP effluents with hydro-morphological alteration was weaker and most pronounced for the frequency of high flow (R² = 0.24) and flow variability (R² = 0.19). About 40 % of the variance in the fish community structure were explained by 12 stressor variables in the CCA models. Water quality and hydrological, but not morphological stressors showed strong albeit highly variable effects on individual fish species. The results indicate that water quality degradation and hydrological alteration are important factors determining the ecological status of fish communities. In this context, WWTP effluents can impose relevant point sources of pollution that affect water quality but also cause alterations of the hydrological regime. Further management measures addressing both stressor groups are needed to improve the ecological status.

Demonstration of an aggregated biomarker response approach to assess the impact of point and diffuse contaminant sources in feral fish in a small river case study.

The assessment of the exposure of aquatic wildlife to complex environmental mixtures of chemicals originating from both point and diffuse sources and evaluating the potential impact thereof constitutes a significant step towards mitigating toxic pressure and the improvement of ecological status. In the current proof-of-concept study, we demonstrate the potential of a novel Aggregated Biomarker Response (ABR) approach involving a comprehensive set of biomarkers to identify complex exposure and impacts on wild brown trout (Salmo trutta fario). Our scenario used a small lowland river in Germany (Holtemme river in the Elbe river catchment) impacted by two wastewater treatment plants (WWTP) and diffuse agricultural runoff as a case study. The trout were collected along a pollution gradient (characterised in a parallel study) in the river. Compared to fish from the reference site upstream of the first WWTP, the trout collected downstream of the WWTPs showed a significant increase in micronucleus formation, phase I and II enzyme activities, and oxidative stress parameters in agreement with increasing exposure to various chemicals. By integrating single biomarker responses into an aggregated biomarker response, the two WWTPs' contribution to the observed toxicity could be clearly differentiated. The ABR results were supported by chemical analyses and whole transcriptome data, which revealed alterations of steroid biosynthesis and associated pathways, including an anti-androgenic effect, as some of the key drivers of the observed toxicity. Overall, this combined approach of in situ biomarker responses complemented with molecular pathway analysis allowed for a comprehensive ecotoxicological assessment of fish along the river. This study provides evidence for specific hazard potentials caused by mixtures of agricultural and WWTP derived chemicals at sublethal concentrations. Using aggregated biomarker responses combined with chemical analyses enabled an evidence-based ranking of sites with different degrees of pollution according to toxic stress and observed effects.

Bioavailability and impacts of estrogenic compounds from suspended sediment on rainbow trout (Oncorhynchus mykiss).

Numerous environmental pollutants have the potential to accumulate in sediments, and among them are endocrine-disrupting chemicals (EDCs). It is well documented that water-borne exposure concentrations of some potent EDCs, more specifically estrogenic- active compounds (ECs), can impair the reproduction of fish. In contrast, little is known about the bioavailability and effects of sediment-associated ECs on fish. Particularly, when sediments are disturbed, e.g., during flood events, chemicals may be released from the sediment and become bioavailable. The main objectives of this study were to evaluate a) whether ECs from the sediment become bioavailable to fish when the sediment is suspended, and b) whether such exposure leads to endocrine responses in fish. Juvenile rainbow trout (Oncorhynchus mykiss) were exposed over 21 days to constantly suspended sediments in the following treatments: i) a contaminated sediment from the Luppe River, representing a "hotspot" for EC accumulation, ii) a reference sediment (exhibiting only background contamination), iii) three dilutions, 2-, 4- and 8-fold of Luppe sediment diluted with the reference sediment, and iv) a water-only control. Measured estrogenic activity using in vitro bioassays as well as target analysis of nonylphenol and estrone via LC-MS/MS in sediment, water, fish plasma, as well as bile samples, confirmed that ECs became bioavailable from the sediment during suspension. ECs were dissolved in the water phase, as indicated by passive samplers, and were readily taken up by the exposed trout. An estrogenic response of fish to Luppe sediment was indicated by increased abundance of transcripts of typical estrogen responsive genes, i.e. vitelline envelope protein α in the liver and vitellogenin induction in the skin mucus. Altered gene expression profiles of trout in response to suspended sediment from the Luppe River suggest that in addition to ECs a number of other contaminants such as dioxins, polychlorinated biphenyls (PCBs) and heavy metals were remobilized during suspension. The results of the present study demonstrated that sediments not only function as a sink for ECs but can turn into a significant source of pollution when sediments are resuspended as during flood-events. This highlights the need for sediment quality criteria considering bioavailability sediment-bound contaminants in context of flood events.

Assessing endocrine disruption in freshwater fish species from a "hotspot" for estrogenic activity in sediment.

Little is known about sediment-bound exposure of fish to endocrine disrupting chemicals (EDC) under field conditions. This study aimed to investigate potential routes of EDC exposure to fish and whether sediment-bound contaminants contribute towards exposure in fish. Tench (Tinca tinca) and roach (Rutilus rutilus) as a benthic and pelagic living fish species, respectively, were sampled at the Luppe River, previously described as a "hotspot" for accumulation of EDC in sediment. A field reference site, the Laucha River, additionally to fish from a commercial fish farm as reference were studied. Blackworms, Lumbriculus variegatus, which are a source of prey for fish, were exposed to sediment of the Luppe River and estrogenic activity of worm tissue was investigated using in vitro bioassays. A 153-fold greater estrogenic activity was measured using in vitro bioassays in sediment of the Luppe River compared the Laucha River. Nonylphenol (NP; 22 mg/kg) was previously identified as one of the main drivers of estrogenic activity in Luppe sediment. Estrogenic activity of Luppe exposed worm tissue (14 ng 17ß-estradiol equivalents/mg) indicated that food might act as secondary source to EDCs. While there were no differences in concentrations of NP in plasma of tench from the Luppe and Laucha, vitellogenin, a biomarker for exposure to EDCs, was induced in male tench and roach from the Luppe River compared to both the Laucha and cultured fish by a factor of 264 and 90, respectively. However, no histological alterations in testis of these fish were observed. Our findings suggest that sediments substantially contribute to the overall EDC exposure of both benthic and pelagic fish but that the exposure did not impact gonad status of the fish.