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To assess the antimicrobial effects of silver ion tubing (ST) on dental unit waterlines (DUWLs) and their sustainability over time. Six dental chair units (DCUs) equipped with ST and four with common tubing (CT) were included in the study. Repeated flushing with phosphate-buffered saline (PBS) was conducted to dislodge biofilms. Then, genetic analysis of the PBS was performed. The tubing was also detached and scanned under a scanning electron microscope (SEM) to observe the adherent biofilm on the lumen walls. Low bacterial levels were