Cross-regional scale pollution of freshwater biofilms unveiled by antibiotic resistance genes.

A comprehensive global profile of the distribution of ARGs in freshwater biofilms is lacking. We utilized metagenomic approaches to reveal the diversity, abundance, transferability and hosts of ARGs in 96 freshwater biofilm samples from 38 sampling sites across four countries. The abundant ARGs were associated with bacitracin, multidrug, polymyxin macrolide-lincosamide-streptogramin (MLS) aminoglycoside, β-lactam, chloramphenicol, sulfonamide and tetracycline resistance, consistent with the spectrum of antibiotics commonly used in human or veterinary medicine. As expected, the resistome in freshwater biofilm habitats was significantly influenced by geographical location and human footprint. Based on the co-occurrence pattern revealed by network analysis, mdtC, kdpE, and emrB were proposed as ARG indicators in freshwater biofilms that can be used to evaluate the abundance of 46 other co-occurring ARG subtypes quantitatively. Metagenomic assembly analysis revealed that the identified ARGs were hosted by more than 46 bacterial phyla, including various pathogens, which greatly expands the knowledge of resistome diversity in freshwater biofilms. Our study points to the central roles of biofilms in harbouring ARGs. The results could enhance understanding the distribution of ARGs in freshwater habitats, thereby strengthening the global environmental risk assessment and management of ARGs.