Leaf litter breakdown is a fundamental process in aquatic ecosystems that is realized by microbial decomposers and invertebrate detritivores. Although this process may be adversely affected by fungicides, among other factors, no test design exists to assess combined effects on such decomposer-detritivore systems. Hence, the present study assessed effects of the model fungicide tebuconazole (65 µg/L) on the conditioning of leaf material (by characterizing the associated microbial community) as well as the combined effects (i.e., direct toxicity and food quality-related effects (=indirect)) on the energy processing of the leaf-shredding amphipod Gammarus fossarum using a five-week semistatic test design. Gammarids exposed to tebuconazole produced significantly less feces (≈ 20%), which in turn significantly increased their assimilation (≈ 30%). Moreover, a significantly reduced lipid content (≈ 20%) indicated lower physiological fitness. The conditioning process was altered as well, which was indicated by a significantly reduced fungal biomass (≈ 40%) and sporulation (≈ 30%) associated with the leaf material. These results suggest that tebuconazole affects both components of the investigated decomposer-detritivore system. However, adverse effects on the level of detritivores cannot be explicitly attributed to direct or indirect pathways. Nevertheless, as the endpoints assessed are directly related to leaf litter breakdown and associated energy transfer processes, the protectiveness of environmental risk assessment for this ecosystem function may be more realistically assessed in future studies by using this or comparable test designs.

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The level of protection provided by the present environmental risk assessment (ERA) of fungicides in the European Union for fungi is unknown. Therefore, we assessed the structural and functional implications of five fungicides with different modes of action (azoxystrobin, carbendazim, cyprodinil, quinoxyfen, and tebuconazole) individually and in mixture on communities of aquatic hyphomycetes. This is a polyphyletic group of fungi containing key drivers in the breakdown of leaf litter, governing both microbial leaf decomposition and the palatability of leaves for leaf-shredding macroinvertebrates. All fungicides impaired leaf palatability to the leaf-shredder Gammarus fossarum and caused structural changes in fungal communities. In addition, all compounds except for quinoxyfen altered microbial leaf decomposition. Our results suggest that the European Union's first-tier ERA provides sufficient protection for the tested fungicides, with the exception of tebuconazole and the mixture, while higher-tier ERA does not provide an adequate level of protection for fungicides in general. Therefore, our results show the need to incorporate aquatic fungi as well as their functions into ERA testing schemes to safeguard the integrity of aquatic ecosystems.

Summary1. Molecular genetic methods continuously uncover cryptic lineages harboured by various species. However, from an applied perspective, it remains unclear whether and to which extent such a genetic diversity affects biological traits (e.g. ecological, behavioural and physiological characteristics) and environmental management. 2. We assessed potential deviations regarding the trait 'environmental stress tolerance' using individuals from five field populations of each of two cryptic lineages (called A and B) comprised under the nominal species Gammarus fossarum. We used ammonia as a chemical stressor while assessing the feeding rate on leaf discs as a measure of sublethal response. In this context, we established a restriction fragment length polymorphism assay to allow a rapid identification of the lineages. 3. We observed a biologically meaningful and statistically significant twofold higher overall tolerance of one cryptic lineage, lineage B, over the other. Confounding factors that may have the potential to influence the test results, such as life stage, sex, season of collection, parasitism, physiological status of organisms and upstream land-use patterns of the river catchments, were either controlled for or displayed only minor deviations between lineages. 4. Synthesis and applications. The trait differences observed in the present study seem to be mainly explained by the considerable genetic differentiation between cryptic lineages of one nominal species. Although traits other than tolerance have been minimally investigated in this context, this study indicates implications in the reliability and quality of environmental monitoring and management if cryptic lineage complexes are ignored.