Abstract：As a pivotal reservoir of antibiotic resistance genes （ARGs）， water environment has become an ideal place for the spread of ARGs and generation of new ARGs. But until now， information about characteristics and functions of phage-borne ARGs in aquatic environments is poorly defined. Here viral metagenomics was used to investigate novel phage-brone ARGs in Nansi Lake and Dongping Lake and then the antibiotic resistance profile of novel phage-borne ARGs was determined by in vitro experiments. The results showed that different classes of ARGs were identified from viromes through analyzing viral reads and contigs， including AAC（6'）， QnrA， VanY， Vat， and β-lactamase genes. More importantly， two novel phage-carried genes encoding β-lactamase， blaNSDPV-1 and blaNSVM-1， were identified by sequence analysis， and the minimum inhibitory concentration test showed that both blaNSDPV-1 and blaNSVM-1 displayed resistance to cephalosporins and carbapenems. In conclusion， these findings indicate that water-borne phages are a reservoir of ARGs or even novel ARGs， therefore playing an important role in the dissemination of ARGs. In addition， viral metagenomics combined with in vitro experiments is an important method to discover novel phage-borne ARGs.