Distinct effects of hypochlorite types on the reduction of antibiotic resistance genes during waste activated sludge fermentation: Insights of bacterial community, cellular activity, and genetic expression

The effectiveness of hypochlorites (NaClO and Ca(ClO)₂) on the reduction of antibiotic resistance genes (ARGs) during waste activated sludge (WAS) fermentation was determined by the quantitative PCR. NaClO and Ca(ClO)₂ exhibited distinct effects on ARGs fates. Ca(ClO)₂ was effective in removing all investigated ARGs, and the efficiency was highly dose-dependent. Unexpectedly, the NaClO treatment attenuated ARGs with lower efficiency and even caused the propagation of certain ARGs (i.e., aadA1 and tetQ) at higher doses. The extracellular polymeric substances dissolution and membrane integrity suggested that unstable NaClO had acute effects on bacteria initially, while it was ineffective to further attenuate ARGs released from hosts due to the rapid consumption of oxidative ClO^-. Without lasting and strong oxidative stress, the microbial activities of tolerant ARGs hosts will partially recover and then contribute to the ARGs dissemination across genera. In contrast, solid-state Ca(ClO)₂ was slowly released and exhibited prolonged effects on bacteria by disrupting cell membranes and removing the susceptible ARGs released from hosts. Furthermore, bacterial taxa-ARG network analysis indicated that Ca(ClO)₂ reduced the abundance of potential hosts, and the metabolic pathway and gene expression related to ARGs propagation were significantly downregulated by Ca(ClO)₂, which contributed to efficient ARGs attenuation.