Water was sampled from source water, the 3-in-1 syringe and the air rotor water line of dental unit water systems (DUWS) in general dental practice in Attica, Greece. A section of the water line supplying the 3-in-1 syringe was cut for biofilm analysis. High total viable counts, Mycobacterium spp. and Pseudomonas aeruginosa were detected in the samples. Back siphonage was evidenced by the presence of blood and the recovery of oral anaerobes from the samples. Legionella pneumophila was recovered from 16.1% of biofilm samples. All tested disinfectants reduced the total viable counts of the DUWS to <100 CFU/mL.

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