Heatwaves, elevated temperatures, and a pesticide cause interactive effects on multi-trophic levels of a freshwater ecosystem.

Climate impacts of elevated temperatures and more severe and frequent weather extremes like heatwaves are globally becoming discernible on nature. While a mechanistic understanding is pivotal for ecosystem management, stressors like pesticides may interact with warming, leading to unpredictable effects on freshwater ecosystems. These multiple stressor studies are scarce and experimental designs often lack environmental realism. To investigate the multiple stressor effects, we conducted a microcosm experiment for 48 days comprising benthic macroinvertebrates, zooplankton, phytoplankton, macrophytes, and microbes. The fungicide carbendazim (100 μg/L) was investigated combined with temperature scenarios representing elevated temperatures (+4 °C) or heatwaves (+0 to +8 °C), both applied with similar energy input on a daily fluctuating ambient temperature (18 °C ± 1.5 °C), which served as control. Measurements showed the highest carbendazim dissipation in water under heatwaves followed by elevated and ambient temperatures. Average carbendazim concentrations were about 50% in water and 16% in sediment of the nominal concentration. In both heated cosms, zooplankton community dynamics revealed an unexpected shift from Rotifera to Cladocera and Copepoda nauplii, indicating variations in their thermal sensitivity, tolerance and resilience. Notably, warming and heatwaves shaped community responses similarly, suggesting heat intensity rather than distribution patterns determined the community structure. Heatwaves led to significant early and longer-lasting adverse effects that were exacerbated over time with Cladocera and Copepoda being most sensitive likely due to significant carbendazim interactions. Finally, a structural equation model demonstrated significant relationships between zooplankton and macrophytes and significantly negative carbendazim effects on zooplankton, whereas positive on macroinvertebrate abundances. The relationship between macroinvertebrate feeding and abundance was masked by significantly temperature-affected microbial leaf litter decomposition. Despite the thermal tolerance of zooplankton communities, our study highlights an increased pesticide threat under temperature extremes. More intense heatwaves are thus likely to cause significant alterations in community assemblages which will adversely affect ecosystem's processes and functions.