Abstract
Broad-host-range plasmids are frequently associated with antibiotic resistance genes and can quickly spread antibiotic resistant phenotypes among diverse bacterial populations. Wastewater treatment plants have been identified as reservoirs for broad-host-range plasmids carrying resistance genes. The threat of broad-host-range plasmids released into the environment from wastewater treatment plants has identified the need for disinfection protocols to target broad-host- range plasmid destruction. Here we evaluate the efficacy of dissolved ozone at 2 and 8 mg·L–1 as a primary means for the destruction of broad-host-range plasmid and chromosomal DNA in simulated effluent. Pilot-scale tests using an experimental unit were carried out in municipal wastewater treatment plant effluent and compared with ultraviolet (UV)-irradiation and chlorination methodologies. Genes specific to Escherichia coli (uidA) and IncP broad-host-range plasmids (trfA) were monitored using real-time quantitative polymerase chain reaction (qPCR), and total DNA was monitored using absorbance spectroscopy. In wastewater treatment plant experiments, E. coli qPCR results were compared to a recognized culture-based method (Colilert?) for E. coli. In laboratory experiments, dissolved ozone at 8 mg·L–1 significantly destroyed 93% total, 98% E. coli, and 99% of broad-host-range plasmid DNA. Ozonation, UV-irradiation, and chlorination significantly reduced DNA concentrations and culturable E. coli in wastewater treat- ment plant effluent. Chlorination and UV disinfection resulted in 3-log decreases in culture-based E. coli concentrations in wastewater treatment plant effluent while changes were not significant when measured with qPCR. Only ozonation significantly decreased the IncP broad-host-range plasmid trfA gene, although concentrations of 2.2 × 105 copies trfA·L–1 remained in effluent. Disinfection processes utilizing high dissolved ozone concentrations for the destruction of emerging contaminants such as broad-host-range plasmid and total DNA may have utility as methods to ensure downstream environmental health and safe water reuse become more important.
Highlights
Municipal and regional wastewater treatment facilities remain a cornerstone of urban and economic development
Chlorination and UV disinfection resulted in 3-log decreases in culture-based E. coli concentrations in wastewater treatment plant effluent while changes were not significant when measured with quantitative polymerase chain reaction (qPCR)
We found that only the higher 8.0 mg·L–1 dose of dissolved ozone significantly reduced total DNA, E. coli chromosomal DNA, and IncP broadhost-range plasmid DNA in 1-L simulated wastewater samples containing dissolved organic carbon to provide a background O3 demand (Table 1)
Summary
Municipal and regional wastewater treatment facilities remain a cornerstone of urban and economic development. Of developing concern are bioreactive compounds that are not targeted by regulations governing discharge of wastewater. This emerging group of contaminants includes compounds such as antibiotic resistance genes, disinfection by-products, pharmaceuticals, and hormones [5,6,7]. Biological contaminants pose a special set of threats among the compounds leaving wastewater treatment plants [8]. A less obvious threat is genetic determinants that can provide antimicrobial resistance phenotypes to pathogenic populations, such as antibiotic resistance genes contained on bacterial plasmids and other mobile genetic elements [1,9,10]. As human population density increases, drinking water resources are increasingly likely
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