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Analysis on Cu bioavailability revealed that extractable Cu2+ was higher in CuSO4-spiked soils than nCuO-spiked soils. At a nCuO exposure concentration of ≤265 mg Cu kg-1 soil, stimulatory effects were observed in nitrification, β-glucosidase and community level physiological profiling (CLPP) tests. nCuO showed no significant inhibitory effects on the soil microbial growth, activity or diversity at the highest concentration (i.e. 627 mg Cu kg-1 soil), with the exception of the dehydrogenase (i.e. ≥27 mg Cu kg-1 soil) and phosphatase (i.