Background: Healthcare-associated infections remain a major public health challenge, with multidrug-resistant organisms (MDROs) accounting for a substantial proportion of cases. Although transmission is traditionally attributed to contact with contaminated surfaces and medical devices, increasing evidence indicates that hospital plumbing systems, particularly handwash basins, may serve as overlooked reservoirs and potential sources of airborne dissemination. Despite this emerging concern, the contribution of handwash basins to bioaerosol generation under real-world conditions remains poorly characterized. To address this gap, we conducted aerosol and surface sampling in five inpatient hospital rooms and three public restrooms.  Methods: Aerosols were collected under two conditions: with the faucet turned off, and turned on, using a single-stage Andersen Cascade Impactor and a BioSpot-VIVAS sampler. Swab samples were also collected from handwash basin drains. All samples were cultured on selective and non-selective agar media, and isolates were identified using MALDI-TOF mass spectrometry, with a focus on carbapenem-resistant bacteria and Candida species.  Results: In inpatient rooms, we detected at least seven MDRO in aerosol samples, including carbapenem-resistant gram-negative bacteria, with significant overlap between swab and aerosol samples supporting splash-mediated aerosolization. Microbial recovery differed substantially between the two sampling methods, demonstrating the limitation of relying on a single aerosol sampling method to characterize airborne microbiota. Conclusion: Our findings indicate that faucet operation in handwash basins can generate aerosols containing MDROs in both clinical and non-clinical settings. Overall, these results identify handwash basins as a potential source contributing to airborne transmission of MDRO, warranting consideration in infection prevention.