Antibiotic resistance spread potential in urban wastewater effluents disinfected by UV/H2O2 process

Abstract

Urban wastewater treatment plants (UWTPs) are among the main hotspots of antibiotic resistance (AR) spread into the environment and the role of conventional and new disinfection processes as possible barrier to minimise the risk for AR transfer is presently under investigation. Accordingly, the aim of this work was to evaluate the effect of an advanced oxidation process (AOP) (specifically UV/H2O2) on AR transfer potential. UV/H2O2 disinfection experiments were carried out on real wastewater samples to evaluate the: i) inactivation of total coliforms, Escherichia coli and antibiotic resistant E. coli as well as ii) possible removal of target antibiotic resistance genes (ARGs) (namely, blaTEM, qnrS and tetW). In particular, DNA was extracted from both antibiotic resistant E. coli bacterial cells (intracellular DNA), grown on selective culture media, and the whole water suspension (total DNA) collected at different treatment times. Polymerase chain reaction (PCR) assay was performed to detect the absence/presence of the selected ARGs. Real Time quantitative Polymerase Chain Reaction (qPCR) was used to quantify the investigated ARGs in terms of copiesmL(-1). In spite of the bacterial inactivation and a decrease of ARGs in intracellular DNA after 60min treatment, UV/H2O2 process was not effective in ARGs removal from water suspension (total DNA). Particularly, an increase up to 3.7×10(3)copiesmL(-1) (p>0.05) of blaTEM gene was observed in total DNA after 240min treatment, while no difference (p>0.05) was found for qnrS gene between the initial (5.1×10(4)copiesmL(-1)) and the final sample (4.3×10(4)copiesmL(-1)). On the base of the achieved results, the investigated disinfection process may not be effective in minimising AR spread potential into the environment. The death of bacterial cells, which results in DNA release in the treated water, may pose a risk for AR transfer to other bacteria present in the receiving water body.

Keywords: Advanced oxidation processes; Antibiotic resistance genes; Antibiotic resistant bacteria; DNA; qPCR.