Given the rapidly increasing use of metal oxide nanoparticles in agriculture as well as their inadvertent addition through sewage sludge application to soils, it is imperative to assess their possible toxic effects on soil functions that are vital for healthy crop production. In this regard, we designed a lab study to investigate the potential toxicity of one of the most produced nanoparticles, i.e. zinc oxide nanoparticles (nZnO), in a calcareous soil. Microcosms of 80 g of dry-equivalent fresh soils were incubated in mason jars for 64 days, after adding 100 or 1000 mg of biogenically produced nZnO kg soil. Moreover, we also added rice-straw derived biochar at 1 or 5% (w: w basis) hypothesizing that the biochar would alleviate nZnO-induced toxicity given that it has been shown to adsorb and detoxify heavy metals in soils. We found that the nZnO decreased microbial biomass carbon by 27.0 to 33.5% in 100 mg nZnO kg soil and by 39.0 to 43.3% in 1000 mg nZnO kg soil treatments across biochar treatments in the short term i.e. 24 days after incubation. However, this decrease disappeared after 64 days of incubation and the microbial biomass in nZnO amended soils were similar to that in control soils. This shows that the toxicity of nZnO in the studied soil was ephemeral and transient which was overcome by the soil itself in a couple of months. This is also supported by the fact that the nZnO induced higher cumulative C mineralization (i.e. soil respiration) at both rates of addition. The treatment 100 mg nZnO kg soil induced 166 to 207%, while 1000 mg nZnO kg soil induced 136 to 171% higher cumulative C mineralization across biochar treatments by the end of the experiment. However, contrary to our hypothesis increasing the nZnO addition from 100 to 1000 mg nZnO kg soil did not cause additional decrease in microbial biomass nor induced higher C mineralization. Moreover, the biochar did not alleviate even the ephemeral toxicity that was observed after 24d of incubation. Based on overall results, we conclude that the studied soil can function without impairment even at 1000 mg kg concentration of nZnO in it.