Response of submerged macrophytes and biofilms to coexisting azithromycin and tetracycline: Antibiotic resistance genes removal, toxicity assessment and microbial properties

Antibiotics, such as azithromycin (AZ), tetracycline (TC), and their related antibiotic resistance genes (ARGs), create serious ecological risks to aquatic organisms. This study examined the response mechanisms of submerged macrophytes and periphytic biofilms to a mixture of AZ and TC pollution and determined the antibiotic removal efficiencies and fate of ARGs. The results showed that the plant-biofilm system had a significant capacity for removing both single and combined antibiotics with removal efficiencies of 93.06% ∼99.80% for AZ and 73.35% ∼97.74% for TC. Higher ARG (tetA, tetC, tetW, ermF, ermX, and ermB) abundances were observed in the biofilm, and subsequent exposure to the antibiotic mixture increased the abundances of these genes. Both single and combined antibiotics triggered antioxidant stress, but antagonistic effects were induced only with mixed AZ and TC exposure. Furthermore, the antibiotics changed the structural characteristics of extracellular polysaccharides and induced alterations in the structure of the biofilm microbial community. Increased N-acylated-l-homoserine lactone confirmed alternations in microbial quorum-sensing. The results extend the understanding of the fate of antibiotics and ARGs when aquatic plants and biofilms are exposed to antibiotic mixtures, as well as the organism's response mechanisms.