Metagenomic analysis of the dichotomous role of uranium in regulating intracellular and extracellular antibiotic resistance genes in activated sludge

Antibiotic resistance genes (ARGs) in activated sludge include intracellular ARGs (iARGs) and extracellular ARGs (eARGs), both of which are recognized as emerging pollutants. While the activated sludge process has been commonly considered for treating wastewater contaminated with radionuclide, the effects and mechanisms of radioactive heavy metals on the fate of iARGs and eARGs (i/e-ARGs) in activated sludge are largely elusive. Here, the distribution, mobility, and hosts of i/e-ARGs in activated sludge during environmental concentrations (50 μg/L and 5000 μg/L) of radioactive uranium (U) stress were explored via metagenomics. The results revealed that the total relative abundance of iARGs and eARGs decreased by 11.62% and 10.41%, respectively, after 90 days of 50 μg/L of U treatment. In contrast, both i/e-multi- and tetracycline ARGs remarkably increased after being exposed to 5000 μg/L of U. Additionally, exposure to 5000 μg/L of U triggered notable decrease in i/e-insertion sequences and plasmids abundance, but significantly enriched i/e-integrons (p < 0.05). Partial least squares pathway modelling indicated that the prevalence of iARGs and eARGs in activated sludge was primarily driven by bacterial hosts and functional genes, respectively. Our findings revealed the dichotomous variation landscape and mechanisms of i/e-ARGs dynamics in activated sludge during U exposure, offering valuable insights for controlling ARGs risk during radioactive wastewater treatment.