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Trimethoprim (TMP) is an antibiotic frequently detected in various environments.Microorganisms are the main drivers of emerging antibiotic contaminant degradation in the environment.However,the feasibility and stability of the anaerobic biodegradation of TMP with sulfate as an electron acceptor remain poorly understood.Here,TMP-degrading microbial consortia were successfully enriched from municipal activated sludge (AS) and river sediment (RS) as the initial inoculums.The acclimated consortia were capable of transforming TMP through demethylation,and the hydroxyl-substituted demethylated product (4-desmethyl-TMP) was further degraded.The biodegradation of TMP followed a 3-parameter sigmoid kinetic model.The potential degraders (Acetobacterium,Desulfovibrio,Desulfobulbus,and unidentified Peptococcaceae) and fermenters (Lentimicrobium and Petrimonas) were significantly enriched in the acclimated consortia.The AS-and RS-acclimated TMP-degrading consortia had similar core microbiomes.The anaerobic biodegradation of TMP could be coupled with sulfate respiration,which gives new insights into the antibiotic fate in real environments and provides a new route for the bioremediation of antibiotic-contaminated environments.