Mesophilic and thermophilic anaerobic digestion of swine manure with sulfamethoxazole and norfloxacin: Dynamics of microbial communities and evolution of resistance genes

The role of norfloxacin (NOR) and sulfamethoxazole (SMX) in mesophilic and thermophilic anaerobic digestion (AD) of pig manure, with respect to methane production and variations in the microbial community and resistance genes, including antibiotic resistance genes (ARGs), class I integrase (intIl), and heavy metal resistance genes (MRGs), was investigated. The results indicated that NOR exerted little influence on the microbial community, whereas SMX negatively affected the acetoclastic methanogens. The abundance of two sulfonamide resistance genes (sul1 and sul2), three quinolone resistance genes (qnrS, parC, and aac(6′)-Ib-cr), and intI1 decreased by 2–3 orders of magnitude at the end of thermophilic AD. In contrast, mesophilic AD was generally ineffective in reducing the abundance of resistance genes. According to the results of redundancy analysis, the abundance of ARGs was affected primarily by microbial community dynamics (68.5%), rather than the selective pressure due to antibiotic addition (13.3%). Horizontal gene transfer (HGT) through intI1 contributed to 26.4% of the ARG variation. The archaeal community also influenced the changes in the resistance genes, and ARG reduction was significantly correlated with enhanced methane production. Thermophilic AD presented a higher methane production potential and greater reduction in resistance gene abundance.