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The soil columns were dismantled and seven soil layers were sampled and analyzed at the end of the second cycle (total experiment time 12 weeks). X-ray absorption spectroscopy analyses were then conducted to assess Zn speciation in OW and OW-amended soils. The results of this study highlighted that (i) the fate of Zn in water-soil-plant compartments was similar, regardless of the type of soil and OW, (ii) 97.6% of the Zn input from OW accumulated in the soil surface layer, (iii) Zn uptake by lettuce increased with repeated OW applica