Vertebrate biodiversity is rapidly decreasing worldwide with amphibians being the most endangered vertebrate group. In the EU, 21 of 89 amphibian species are recognized as being endangered. The intensively used European agricultural landscape is one of the major causes for these declines. As agriculture represents an essential habitat for amphibians, exposure to pesticides can have adverse effects on amphibian populations. Currently, the European risk assessment of pesticides for vertebrates requires specific approaches for fish regarding aquatic vertebrate toxicity and birds as well as mammals for terrestrial vertebrate toxicity but does not address the unique characteristics of amphibians. Therefore, the overall goal of this thesis was to investigate the ecotoxicological effects of pesticides on Central European anuran amphibians. For this, effects on aquatic and terrestrial amphibian life stages as well as on reproduction were investigated. Then, in anticipation of a risk assessment of pesticides for amphibians, this thesis discussed potential regulatory risk assessment approaches.
For the investigated pesticides and amphibian species, it was observed that the acute aquatic toxicity of pesticides can be addressed using the existing aquatic risk assessment approach based on fish toxicity data. However, lethal as well as sublethal effects were observed in terrestrial juveniles after dermal exposure to environmentally realistic pesticide concentrations, which cannot be covered using an existing risk assessment approach. Therefore, pesticides should also be evaluated for potential terrestrial toxicity using risk assessment tools before approval. Additionally, effects of co-formulants and adjuvants of pesticides need specific consideration in a future risk assessment as they can increase toxicity of pesticides to aquatic and terrestrial amphibian stages. The chronic duration of combined aquatic and terrestrial exposure was shown to affect amphibian reproduction. Currently, such effects cannot be captured by the existing risk assessment as data involving field scenarios analysing effects of multiple pesticides on amphibian reproduction are too rare to allow comparison to data of other terrestrial vertebrates such as birds and mammals. In the light of these findings, future research should not only address acute and lethal effects, but also chronic and sublethal effects on a population level. As pesticide exposure can adversely affect amphibian populations, their application should be considered even more carefully to avoid further amphibian declines. Overall, this thesis emphasizes the urgent need for a protective pesticide risk assessment for amphibians to preserve and promote stable amphibian populations in agricultural landscapes.
Studies have shown that wastewater treatment plant (WWTP) effluents are the major pathways of organic and inorganic chemicals of anthropogenic use (=micropollutants) into aquatic environments. There, micropollutants can be transferred to ground water bodies - and may finally end up in drinking water - or cause various effects in aquatic organisms like multiple resistances of bacteria. Hence, the upgrading of WWTPs with the aim to reduce the load of those micropollutants is currently under discussion.
Therefore, the primary objective of this thesis was to assess ecotoxicological effects of wastewater ozonation, a tertiary treatment method, using specifically developed toxicity tests with Gammarus fossarum (Koch) at various levels of ecological complexity. Several studies were designed in the laboratory and under semi-field conditions to cope with this primary objective. Prior to the investigations with ozone treated wastewater, the ecotoxicity of secondary treated (=non-ozone treated) wastewater from WWTP Wüeri, Switzerland, for the test species was assessed by a four-week experiment. This experiment displayed statistically significant impairments in feeding, assimilation and physiological endpoints related to population development and reproduction. The first experiment investigating ecotoxicological implications of ozone application in wastewater from the same WWTP displayed a preference of G. fossarum for leaf discs conditioned in ozone treated wastewater when offered together with leaf discs conditioned in non-ozone treated wastewater. This effect seems to be mainly driven by an alteration in the leaf associated microbial community. Another series of laboratory experiments conducted also with wastewater from WWTP Wüeri treated with ozone at the lab- or full-scale, revealed significantly increased feeding rates of G. fossarum exposed to ozone treated wastewater compared to non-ozone treated wastewater. These laboratory experiments also indicated that any alteration in the organic matrix potentially caused by ozone treatment is not related to the effects in feeding as this endpoint showed only negligible deviation in secondary treated wastewater, which contained hardly any (micro)pollutants (i.e. pharmaceuticals), from the same wastewater additionally treated with ozone. Moreover, it was shown that shifts in the dissolved organic carbon (DOC) profile do not affect the feeding rate of gammarids. In situ bioassays conducted in the receiving stream of the WWTP Wüeri confirmed the results of the laboratory experiments by displaying significantly reduced feeding rates of G. fossarum exposed below the WWTP effluent if non-ozone treated wastewater was released. However, at the time the ozonation was operating, no adverse effects in feeding rates were observed below the effluent compared to the unaffected upstream sites. Also population studies in on-site flow-through stream microcosms displayed an increased feeding and a statistically significantly higher population size after ten weeks when exposed to ozone treated wastewater compared to non-ozone treated wastewater.
In conclusion, the present thesis documents that ozonation might be a suitable tool to reduce both the load of micropollutants as well as the ecotoxicity of wastewaters. Thus, this technology may help to meet the requirements of the Water Framework Directive also under predicted climate change scenarios, which may lead to elevated proportions of wastewater in the receiving stream during summer discharge. However, as ozone application may also produce by-products with a higher toxicity than their parent compounds, the implementation of this technique should be assessed further both via chemical analysis and ecotoxicological bioassays.