Importance of sludge conditioning in attenuating antibiotic resistance: Removal of antibiotic resistance genes by bioleaching and chemical conditioning with Fe[III]/CaO.

Conditioning can drastically improve the dewaterability of sewage sludge and thus it is widely practiced in most wastewater treatment plants (WWTPs). In WWTPs, various antibiotic resistance genes (ARGs) present in sewage are concentrated in the sewage sludge, but the effect of sludge conditioning on ARGs in sewage sludge remains unclear. Here, we evaluated and compared the effectiveness of four sludge conditioning methods (namely chemical conditioning with polyacrylamide (PAM), chemical conditioning with Fe[III]/CaO, bioleaching conditioning, and chemical acidification conditioning) and an aerobic incubation control in removing 46 target ARGs and intI1 from a municipal sewage sludge. The damage of sludge microbial cells and the change in the sludge bacterial community during the various sludge conditioning treatments were also characterized. The results suggested that the chemical conditioning with PAM and aerobic incubation treatment did not remove ARGs and intI1 from the sewage sludge. The chemical acidification reduced the absolute abundances of most ARGs and intI1, but increased their relative abundances. However, the chemical conditioning with Fe[III]/CaO and bioleaching conditioning reduced both the absolute and relative abundances of most ARGs and removed a majority of extracellular ARGs in the sludge. During sludge conditioning treatments, the sludge microbial cells were severely damaged to decrease the total bacterial biomass in sludge, and accordingly the bacterial hosts carrying ARGs and intI were effectively damaged to reduce the absolute abundances of most ARGs and intI1. In addition, the sludge bacterial community in conditioned sludge determined the relative abundances of residual ARGs. Our findings suggest that sludge conditioning can be an important sludge treatment process in attenuating antibiotic resistance in sewage sludge, and bioleaching and chemical conditioning with Fe[III]/CaO can be employed as effective conditioning ways to reduce ARGs in sewage sludge, potentially limiting their release to the environment.