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Acute toxicity and genotoxic activity of 11 pollutants were investigated in wild populations of Diamesa cinerella and Diamesa zernyi (Diptera Chironomidae) from two alpine streams (Italian Alps). D. cinerella was collected in two sites on the non-glacial Vermigliana stream, 50 m-upstream and 5-m downstream of the Wastewater Treatment Plant (WTP) at the Tonal Pass (1799 m a.s.l.). D. zernyi was collected in the Presena glacial stream, close to the glacier snout (2685 m a.s.l.). IV-instar larvae were exposed for 24-96 h to increasing concentrations of three drugs (ibuprofen-IBU, furosemide-FUR, trimethoprim-TMP), three personal care products (triclocarban-TCC, tonalid-TON, sucralose-SUCR), and five pesticides (boscalid-BOS, captan-CAP, chlorpyrifos-CPS, metolachlor-MET, terbuthylazine-TER). The experimental concentrations were from one to several million times higher than the highest environmental concentration (EC) measured in the study sites. Two mixtures of pesticides were also prepared: MIX 1K =103 x EC of CPS, MET and TER, and MIX 10K = 104 x EC of CPS, MET and TER. Species- and site-specific responses were observed for both tests. On the basis of survival data, both species resulted very resistant to pharmaceuticals (mainly to FUR for which no effects on survival and movement or pupation were observed), and more sensitive to pesticides (mainly to CPS, MET and CAP). Genotoxicity tests (Comet assay) highlighted a WTP effect under natural conditions and a genotoxic effect for 9 of the 11 tested compounds. Overall, a clear gradient of increasing resistance in larvae from the least (PR0) to the most polluted (TP_dw) site was highlighted by both tests, ecotoxicological and of genotoxicity, as also expected according to species autecology (D. zernyi is restricted to very cold and pristine habitats). D. cinerella living downstream of the effluent accumulates a significantly higher DNA damage than the other populations, highlighting a basal physiological stress condition in nature. It is plausible that these larvae possess chemical resistance strategies to survive already under natural conditions. Diamesa spp. exhibited a higher toxic resistance than any other model species tested to date under the same pollutants, probably associable to its strong cold resistance. The results emphasised that the measured concentrations of Contaminants of Emerging Concern (CECs) and pesticides seem to be far below those required to cause acute effects. However, the effects on freshwater communities of prolonged exposure to mixture of trace CECs and pesticides remain unknown.