Individual performances and biochemical pathways as altered by field-realistic exposures of current-use fungicides and their mixtures in a non-target species, Gammarus fossarum.

Persistent fungicides, which are widely applied to agricultural soils to protect crops, are frequently detected in freshwaters because of hydraulic transfer, possibly resulting in unintentional adverse effects on wildlife. However, the ecotoxicity of fungicides in aquatic species remains scarcely assessed at environmentally relevant concentrations, and there is scant information available on their combined effects. This study aims at investigating multi-level changes elicited by two currently used fungicides, boscalid and tebuconazole, in the amphipod Gammarus fossarum. In microcosms, gammarids were exposed for 72 h to fungicides tested individually or in binary mixture at 0.01, 0.1 and 1.0 μg/L to monitor individual performances (locomotion, respiration and amplexus formation) and biochemical parameters (involved in energy metabolism, growth, moulting and cell stress). This range of exposure concentrations was field-realistic and largely lower than local environmental quality standards for the protection of aquatic wildlife. Overall, results showed that single and combined exposures altered the mobility and respiratory activity of individuals. At the cellular level, boscalid inhibited energy-based biomarkers whereas tebuconazole led to cytotoxicity associated with reduced antioxidant defences. In binary mixtures, the biochemical responses were mainly driven by the presence of boscalid. Multi-variable analyses, integrating individual alterations and cellular impairments alike, confirmed the relevance of the multi-level approach in forecasting the toxicity of fungicides and their mixtures towards other aquatic species. This study demonstrates dose-related sublethal effects of fungicides on multiple functional traits in an ecosystem engineer under realistic exposure scenarios, highlighting the harmful signs of these toxicants. Such sublethal alterations could thus constitute reliable tools for the early diagnosis of the organisms' health and the ecological status of agriculturally impacted hydrosystems.