From the Reclaimed Water Treatment Plant to Irrigation in Intensive Agriculture Farms: Assessment of the Fate of Antibiotics, Antibiotic Resistance Bacteria and Genes, and Microbial Pathogens at Real Scale

This work aims to investigate the occurrence of 31 antibiotics (ABs), 2 bacteria (<i>Escherichia coli</i> and <i>Pseudomonas</i> spp.) and their counterpart antibiotic-resistant bacteria (carbapenem and cephalosporin families), and several antibiotic-resistant genes (ARGs) throughout a full distribution system of reclaimed water (RW) in a real-scale scenario. The RW was analyzed (i) before and after the tertiary treatment (sand filtration and chlorination), (ii) during the storage period in secondary ponds before its use in irrigation, and (iii) directly in the droppers installed in four plastic-based greenhouses over 9 months. The results obtained in RW showed a bacterial concentration below the minimum required to reach class A (<10 CFU/100 mL, Regulation EU 2020/741), a reduction of the initial AB concentration (up to 13 ABs, total 4847 ± 1413 ng/L) of 58%, and no significant reduction of ARGs (Log units/100 mL: 16S rRNA (9.99 ± 0.80) > <i>intI1</i> (8.80 ± 0.95) > <i>bla_CTX-M32</i> (7.53 ± 0.63) > <i>sul1</i> (7.08 ± 1.05) > <i>bla</i>_<i>TEM</i> (6.81 ± 1.05) > <i>qnrS</i> (5.72 ± 0.82)). The storage of RW was a hotspot only for bacteria; an increase in all concentrations was observed in both main and secondary reservoirs, demonstrating that direct RW reuse is the most beneficial option to avoid significant bacterial regrowth. In all greenhouse droppers' systems, a significantly higher concentration of all bacteria was generally detected than in secondary reservoirs, demonstrating that this is another hotspot independent of whether the RW is used directly or not. Therefore, the RW storage and distribution may negatively affect the microbial water quality, while ABs and ARGs are detected along the entire scheme of urban wastewater reclamation and reuse, reaching the greenhouse environment (including soil and plants).