Degradation of extracellular genomic, plasmid DNA and specific antibiotic resistance genes by chlorination

Highlights

Extracellular DNA structure damaged by chlorination was characterized.

Integrity of extracellular ARG genetic information after chlorination was determined.

Typical chlorine doses will likely effectively diminish extracellular DNA and ARGs.

Plasmid DNA/ARGs were less readily broken down than genomic DNA.

The Bioanalyzer methodology effectively documented damage incurred to DNA.

Abstract

There is a need to improve understanding of the effect of chlorine disinfection on antibiotic resistance genes (ARGs) in order to advance relevant drinking water, wastewater, and reuse treatments. However, few studies have explicitly assessed the physical effects on the DNA. Here we examined the effects of free chlorine (1–20 mg Cl₂/L) on extracellular genomic, plasmid DNA and select ARGs. Chlorination was found to decrease the fluorometric signal of extracellular genomic and plasmid DNA (ranging from 0.005 to 0.05 mg/mL) by 70%, relative to a no-chlorine control. Resulting DNA was further subject to a fragment analysis using a Bioanalyzer, indicating that chlorination resulted in fragmentation. Moreover, chlorine also effectively deactivated both chromosomal- and plasmid-borne ARGs, mecA and tetA, respectively. For concentrations >2 mg Cl₂//L × 30 min, chlorine efficiently reduced the qPCR signal when the initial concentration of ARGs was 10⁵ copies/mL or less. Notably, genomic DNA and mecA gene signals were more readily reduced by chlorine than the plasmid-borne tetA gene (by ~2 fold). Based on the results of qPCR with short (~200 bps) and long amplicons (~1200 bps), chlorination could destroy the integrity of ARGs, which likely reduces the possibility of natural transformation. Overall, our findings strongly illustrate that chlorination could be an effective method for inactivating extracellular chromosomal- and plasmid-borne DNA and ARGs.

Graphical abstract

Keywords

Antibiotic resistance / Antibiotic resistance genes (ARGs) / Extracellular DNA/ARGs / Chlorination

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Acknowledgements

This work was supported in part by the National Science Foundation Partnership in International Research and Education (PIRE): OISE Award # 1545756, “Halting Environmental Antimicrobial Resistance Dissemination”, National Key Research and Development Program of China-International collaborative project from Ministry of Science and Technology (No. 2017YFE0107300), and Virginia Tech ICTAS Center for Science and Engineering of the Exposome (SEE) and The Virginia Tech National Center for Earth and Environmental Nanotechnology National Science Foundation Grant (NNCI- 1542100).

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