Removal of Intracellular and Extracellular Antibiotic Resistance Genes and Virulence Factor Genes Using Electricity-Intensified Constructed Wetlands

Constructed wetland (CW) is considered a promising technology for the removal of emerging contaminants. However, its removal performance for antibiotic resistance genes (ARGs) is not efficient and influence of virulence factor genes (VFGs) have not been elucidated. Here, removal of intracellular and extracellular ARGs as well as VFGs by electricity-intensified CWs was comprehensively evaluated. The two electrolysis-intensified CWs can improve the removal of intracellular ARGs and MGEs to 0.96- and 0.85-logs, respectively. But cell-free extracellular ARGs (CF-eARGs) were significantly enriched with 1.8-logs in the electrolysis-intensified CW. Interestingly, adding Fe-C microelectrolysis to the electrolysis-intensified CW is conducive to the reduction of CF-eARGs. However, the detected number and relative abundances of intracellular and extracellular VFGs were increased in all of the three CWs. The biofilms attached onto the substrates and rhizosphere are also hotspots of both intracellular and particle-associated extracellular ARGs and VFGs. Structural equation models and correlation analysis indicated that ARGs and VFGs were significantly cooccurred, suggesting that VFGs may affect the dynamics of ARGs. The phenotypes of VFGs, such as biofilm, may act as protective matrix for ARGs, hindering the removal of resistance genes. Our results provide novel insights into the ecological remediation technologies to enhance the removal of ARGs. Constructed wetland (CW) has been considered a promising technology for the removal of emerging contaminants. But its removal performance for antibiotic resistance genes (ARGs) is controversial. Here, we proposed electrolysis-intensified CW to enhance the removal of intracellular and extracellular ARGs. This strategy significantly promotes the removal of resistance genes. In addition, we evaluated the correlations between virulence factor genes (VFGs) and ARGs, suggesting that VFGs may affected the removal of ARGs using biofilm- or substrate-based technologies. These results lead significant guidance for the enhancement removal of ARGs using CWs, and highlight the need to take VFGs into account.