Ecotoxicological effects of copper oxide nanoparticles (nCuO) on the soil microbial community in a biosolids-amended...

This study represents a holistic approach in assessing the effects of copper oxide nanoparticles (nCuO) on microbial health and community structure in soil amended with municipal biosolids. The biosolids were amended with nCuO (<50 nm) and mixed into a sandy loam soil at measured Cu concentrations of 27, 54, 123, 265 and 627 mg Cu kg^-1 soil. A suite of tests were used to assess the potential impact of nCuO on microbial growth, activity, and diversity. Microbial growth was determined by the heterotrophic plate count (HPC) method, while microbial diversity was assessed using both community level physiological profiling (CLPP) and 16S ribosomal DNA (rDNA) sequencing. Microbial activity was assessed by examining soil nitrification, organic matter decomposition, soil respiration (basal and substrate induced) and soil enzyme assays for dehydrogenase, phosphatase and β-glucosidase activities. As a readily soluble positive control, copper sulfate (CuSO₄) was used at measured Cu concentrations of 65, 140, 335 and 885 mg Cu kg^-1 soil for select tests, and at the highest concentration for the remaining tests. 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.e. 627 mg Cu kg^-1 soil) enzyme activities. In contrast, inhibition from CuSO₄ at 885 mg Cu kg^-1 soil was observed in all tests with the exception of β-glucosidase enzyme activity. The growth of a Cu tolerant bacterium, Rhodanobacter sp., was observed at 885 mg Cu kg^-1 soil (CuSO₄).