Of causes and symptoms: using monitoring data and expert knowledge to diagnose the causes of stream degradation.

Ecological status assessment under the European Water Framework Directive (WFD) often integrates the impact of multiple stressors into a single index value. This hampers the identification of individual stressors being responsible for status deterioration. As a consequence, management measures are often disentangled from assessment results. To close this gap and to support river basin managers in the diagnosis of stressors, we linked numerous macroinvertebrate assessment metrics and one diatom index with potential causes of ecological deterioration through Bayesian belief networks (BBNs). The BBNs were informed by WFD monitoring data as well as regular consultation with experts and allow to estimate the probabilities of individual degradation causes based upon a selection of biological metrics. Macroinvertebrate metrics were shown to be stronger linked to hydromorphological conditions and land use than to water quality-related parameters (e.g., thermal and nutrient pollution). The modeled probabilities also allow to order the potential causes of degradation hierarchically. The comparison of assessment metrics showed that compositional and trait-based community metrics performed equally well in the diagnosis. The testing of the BBNs by experts resulted in an agreement between model output and expert opinion of 17-92% for individual stressors. Overall, the expert-based validation confirmed a good diagnostic potential of the BBNs; on average 80% of the diagnosed causes were in agreement with expert judgement. We conclude that diagnostic BBNs can assist the identification of causes of stream and river degradation and thereby inform the derivation of appropriate management decisions.

A new enzymatic method assessing the impact of wastewater treatment plant effluents on the assimilative capacity of small rivers.

Microorganisms play an important role in maintaining a good water quality in rivers by degrading organic material, including toxic substances. In the present study, we analyzed the potential impact of municipal wastewater treatment plant (WWTP) effluents as a major stress factor on the assimilative capacity of small rivers. It was the aim to develop a new bioassay for assessing such impacts in the receiving rivers by measuring the activity of extracellular enzymes (exoenzymes) in bacteria. Therefore, we established a specific in-vitro assay to detect inhibitory effects of solid phase-enriched water samples on ß-glucosidase (BGL) activity of the actinobacterium Cellulomonas uda as a proxy for the microbial decomposition of organic substances and thus for the assimilative capacity of surface waters. We found significant reductions of BGL activity in the WWTP effluents and in the receiving waters directly downstream as well as a relative quick recovery over the further course of the water bodies. The new bioassay offers a promising tool for the assessment of the assimilative capacity in surface waters and a potential impact of WWTP effluents on this key ecosystem function. Abbreviations WWTP wastewater treatment plant BGL ß-glucosidase EU-WFD European Water Framework Directive FAU Formazin Attenuation Units PE population equivalents REF relative enrichment factor; SPE solid phase extraction MTBE methyl-tert-buthyl-ether DMSO dimethyl-sulfoxide NPG 4-nitrophenol-ß-d-glucopyranoside DOC dissolved organic carbon.

Small but with big impact? Ecotoxicological effects of a municipal wastewater effluent on a small creek.

Municipal wastewater treatment plants (WWTPs) discharge micropollutants like pharmaceuticals, pesticides, personal care products or endocrine disrupting chemicals but also nutrients. Both can adversely influence the freshwater ecosystem and may finally affect the ecological conditions. Many studies focus on the potential impact of large WWTPs even if smaller ones are more common, often less efficient and discharge into small creeks or the upper reaches of rivers. As a result, the receiving waters are characterized by relatively high shares of treated wastewater. Thus, the primary objective of this study was to investigate the ecotoxicological effects of a small WWTP on freshwater amphipods and mollusks in a small creek using an active and passive monitoring approach, accompanied by laboratory experiments (LE). In vitro assays with recombinant yeasts and the microtox assay with Aliivibrio fischeri were performed in parallel to determine the endocrine potential and the baseline toxicity. The evaluation of the effects of the analysed WWTP was possible due to its shutdown during our study and the application of the same in vivo and in vitro assays before and after the shutdown. During the operation of the WWTP the discharge of treated wastewater caused significantly higher mortalities and lower reproduction of the anaylsed invertebrates in the active and passive montoring as well as in the LEs. Furthermore, the amphipod species assemblage in the creek was affected downstream of the WWTP effluent. Besides, the endocrine activity and baseline toxicity were significantly higher downstream of the effluent. After the shutdown of the WWTP, the in vitro activity levels and adverse in vivo effects in the receiving water recovered quickly with no significant differences downstream of the former WWTP effluent compared to the upstream station. Furthermore, the previously disturbed amphipod species assemblage recovered significantly with a shift in favor of Gammarus fossarum downstream of the effluent. These biological results are consistent with a marked decline by 81.5% for the detected micropollutants in the receiving creek after the shutdown which points to a prominent role of micropollutants for the observed effects.

Effects of metoprolol on aquatic invertebrates in artificial indoor streams.

Aquatic organisms are impacted by various biotic and abiotic stressors such as current, inter- and intraspecific competition for food resources and habitat, neobiota as well as an increasing number of chemicals. The latter also include pharmaceuticals, which are increasingly being detected in surface waters due to their growing use. The aim of our study was to determine effect data for metoprolol as a model compound for beta-blockers under an environmentally realistic exposure scenario on aquatic invertebrates inhabiting lotic environments. To this end we performed a 40-day experiment in artificial indoor streams (AIS) located in a greenhouse. We focussed on three autochthonous invertebrate species with high relevance in stream ecology: the amphipod Gammarus fossarum, the gastropod Potamopyrgus antipodarum, and the oligochaete Lumbriculus variegatus. Effects on reproduction were found with EC10 (40 days) values of 0.092 mg L-1 (G. fossarum), 0.253 mg L-1 (P. antipodarum), and 0.596 mg L-1 (L. variegatus). Considering environmental data, metoprolol seems to pose no hazard for aquatic invertebrates at present exposure levels.

Optimizing the design of a reproduction toxicity test with the pond snail Lymnaea stagnalis.

This paper presents the results from two ring-tests addressing the feasibility, robustness and reproducibility of a reproduction toxicity test with the freshwater gastropod Lymnaea stagnalis (RENILYS strain). Sixteen laboratories (from inexperienced to expert laboratories in mollusc testing) from nine countries participated in these ring-tests. Survival and reproduction were evaluated in L. stagnalis exposed to cadmium, tributyltin, prochloraz and trenbolone according to an OECD draft Test Guideline. In total, 49 datasets were analysed to assess the practicability of the proposed experimental protocol, and to estimate the between-laboratory reproducibility of toxicity endpoint values. The statistical analysis of count data (number of clutches or eggs per individual-day) leading to ECx estimation was specifically developed and automated through a free web-interface. Based on a complementary statistical analysis, the optimal test duration was established and the most sensitive and cost-effective reproduction toxicity endpoint was identified, to be used as the core endpoint. This validation process and the resulting optimized protocol were used to consolidate the OECD Test Guideline for the evaluation of reproductive effects of chemicals in L. stagnalis.

Development and validation of an OECD reproductive toxicity test guideline with the pond snail Lymnaea stagnalis (Mollusca, Gastropoda).

The OECD test guideline development program has been extended in 2011 to establish a partial life-cycle protocol for assessing the reproductive toxicity of chemicals to several mollusk species, including the great pond snail Lymnaea stagnalis. In this paper, we summarize the standard draft protocol for a reproduction test with this species, and present inter-comparison results obtained in a 56-day prevalidation ring-test using this protocol. Seven European laboratories performed semi-static tests with cultured snails of the strain Renilys® exposed to nominal concentrations of cadmium chloride (from 53 to 608µgCdL(-1)). Cd concentrations in test solutions were analytically determined to confirm accuracy in the metal exposure concentrations in all laboratories. Physico-chemical and biological validity criteria (namely dissolved oxygen content >60% ASV, water temperature 20±1°C, control snail survival >80% and control snail fecundity >8 egg-masses per snail over the test period) were met in all laboratories which consistently demonstrated the reproductive toxicity of Cd in snails using the proposed draft protocol. Effect concentrations for fecundity after 56days were reproducible between laboratories (68