Pseudomonas aeruginosa is an epitome of opportunistic pathogens. It is the third most common pathogen associated with hospital acquired catheter associated urinary tract infections (CAUTIs). This pathogen forms biofilms on the surface of indwelling catheters which are resistant to antimicrobial agents as well as to host defense mechanisms leading to chronicity and recurrence of infections. In the present study, elaboration of virulence factors by immature and mature biofilm cells of P. aeruginosa was assessed in vitro. It was observed that with increase in time period of biofilm generation from 1 to 4 day (immature biofilms) there was increase in elaboration of majority of virulence traits. However 5 day old biofilms (mature biofilms) were least producers of all the virulence determinants which continued till 7 th day. Implications of these findings in relation to urinary tract infections caused by biofilms of P. aeruginosa have been discussed.