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The quality of dental unit water is of considerable importance since patients and dental staff are regularly exposed to water and aerosols generated from the dental unit waterlines (DUWLs). Pseudomonas aeruginosa contamination in dental unit waterlines was reported by its origin from incoming local water supplies of the dental clinics. A total of 82 dental unit water samples from 34 dental clinics of Amravati city were collected, out of which 59 water samples were contaminated by P. aeruginosa. The ultrasonic scaler showed maximum 81% contamination of P. aeruginosa then other two hand-pieces, 3 in 1 syringe 68% and air rotor 69%. The dental clinics using distilled water as a source of water for treatment showed 72% P. aeruginosa contamination whereas Undergraduate (UG) dentists clinics showed 75% and Postgraduates (PG) dentists clinics showed 68% P. aeruginosa in DUWLs water.