Behavior of antibiotic resistance genes under extremely high-level antibiotic selection pressures in pharmaceutical wastewater treatment plants

Pharmaceutical wastewater treatment plants (PWWTPs), which receive wastewater containing extremely high levels of antibiotics, are regarded as potential hot spots for antibiotic-resistance development in the environment. Six sampling campaigns in six PWWTPs in Southeastern China were carried out to assess the prevalence and fate of antibiotic resistance genes (ARGs). Different genes were monitored in different PWWTPs (PWWTP A: lincosamides; PWWTP B: aminoglycosides and macrolides; PWWTP C: quinolones; PWWTP D: macrolides and quinolones; PWWTP E: cephalosporins; and PWWTP F: quinolones and macrolides) using real-time quantitative polymerase chain reactions (PCRs), according to the antibiotic type produced. The levels of typical ARG subtypes in the final effluents ranged from (1.03±0.91)×101 to (6.78±0.21)×107copies/mL. The absolute abundance of ARGs in effluents accounted for 0%-577% of influents to the six PWWTPs with a median value of 6%. Most of the ARGs are transported to the dewatered sludge, with concentrations from (1.38±0.21)×105 to (6.84±0.43)×1010copies/g dry weight (dw). In different treatment units (before/after biological units), a clear trend of proliferation or attenuation was not observed for the ARGs, aside from a strong attenuation in moving bed bio-film reactor (MBBR) in PWWTP C. Through correlation analyses, this study demonstrated that the bacterial abundance and antibiotic concentrations within the PWWTPs influenced the fate of the associated ARGs, and this was possibly related primarily to the intrinsic resistance mechanisms of corresponding ARGs. Macrolide ARGs, which tend to locate in plasmids and transposons, positively correlate weakly with total macrolide antibiotic concentrations but positively correlate strongly with 16S rRNA concentrations. Furthermore, ARG concentrations in the wastewater from fermentation were significantly higher than in the wastewater from chemical synthesis and preparation. This is the first comprehensive study on the behavior of antibiotic resistance genes under extremely high-level antibiotic selection pressures in pharmaceutical wastewater treatment plants (PWWTPs) in Southeastern China.