Pseudomonas aeruginosa is an opportunistic and nosocomial pathogen associated with respiratory tract infections. Due to the irrational use of antibiotics, P. aeruginosa has developed antibiotic resistance, resulting in increased morbidity and mortality rates. Medicinal plants of reported antibacterial activity can be alternatively used to treat its infections and do not result in resistance. The purpose of our study was to check for the antibiofilm potential of Juglans regia L. (Persian walnut) extracts against waterborne P. aeruginosa. The aqueous and methanol extracts were checked for antibacterial activity by agar well diffusion method. The biofilm formation assay carried out for the plant extracts gave MIC values of 3 and 5 µg/ml for P. aeruginosa strains. The thin layer chromatography (TLC), revealed the active components in the form of two bands. Our study indicated that the native plants of Pakistan have bioactivity against biofilm-forming pathogens.