BACKGROUND: Antimicrobial resistant (AMR) pathogens in drinking water plumbing systems represent a significant yet underestimated public health threat. METHODS: This is the first study to use qPCR and culture-based methods to investigate the prevalence of key AMR threats, methicillin resistant Staphylococcus aureus (MRSA) and carbapenem resistant Pseudomonas aeruginosa and Acinetobacter baumannii, in Australian hospital and residential drinking water and biofilm samples. RESULTS: Seventy three percent of residential water and biofilm samples were qPCR positive for at least one target pathogen compared with 38% of hospital samples, and 45% of residential plumbing fixtures harboured at least two target pathogens. Thirty seven percent of water and biofilm samples were qPCR positive for P. aeruginosa, 22.3% for A. baumannii and 21.7% for S. aureus. Using culture, 10% of samples were positive for P. aeruginosa, 8% for A. baumannii and 7% for S. aureus. Of these, 29% of P. aeruginosa and 28% of A. baumannii culture isolates were carbapenem resistant, and 54% of S. aureus isolates were identified as MRSA. Drain biofilms were the most common reservoir for AMR A. baumannii, S. aureus and P. aeruginosa. Carbapenem resistance genes including bla CONCLUSION: These findings underscore the critical role of plumbing biofilms as hotspots for diverse AMR pathogens, amplifying risks for vulnerable populations, particularly in home healthcare settings. This study highlights an urgent need for enhanced surveillance and targeted interventions to mitigate AMR risks in drinking water plumbing systems.