The study investigates the distribution, abundance, and activity of prophage-encoded antibiotic resistance genes (pARGs) in different habitats, focusing on the impact of human activities on their spread. By analyzing 38,605 bacterial genomes, 1432 metagenomes, and 1186 metatranscriptomes across 12 contrasting habitats, the researchers found a significant increase in the abundance, diversity, and activity of pARGs in human-impacted environments. These environments, characterized by higher antibiotic exposure, showed a higher proportion of lysogenic bacteria and a higher content of pARGs per lysogen. The study also revealed that pARGs from human-impacted habitats were more likely to be transmitted between different environments and bacterial taxa, with a higher transcriptional activity and a higher risk of transmission. Experimental validation using mitomycin C treatment confirmed the induction of prophages and the resistance conferred by pARGs in a subset of tested strains. The findings suggest that human activities alter phage-host interactions, enriching pARGs in prophages and facilitating their global spread, which poses a significant threat to public health.The study investigates the distribution, abundance, and activity of prophage-encoded antibiotic resistance genes (pARGs) in different habitats, focusing on the impact of human activities on their spread. By analyzing 38,605 bacterial genomes, 1432 metagenomes, and 1186 metatranscriptomes across 12 contrasting habitats, the researchers found a significant increase in the abundance, diversity, and activity of pARGs in human-impacted environments. These environments, characterized by higher antibiotic exposure, showed a higher proportion of lysogenic bacteria and a higher content of pARGs per lysogen. The study also revealed that pARGs from human-impacted habitats were more likely to be transmitted between different environments and bacterial taxa, with a higher transcriptional activity and a higher risk of transmission. Experimental validation using mitomycin C treatment confirmed the induction of prophages and the resistance conferred by pARGs in a subset of tested strains. The findings suggest that human activities alter phage-host interactions, enriching pARGs in prophages and facilitating their global spread, which poses a significant threat to public health.