Non-albicans Candida yeasts are incriminated in systemic mycoses, and are often found with bacteria in multi-species biofilms. The management of biofilm-related infections is difficult and new therapeutic solutions are therefore necessary. The aim of this study is to evaluate the effectiveness of the antimicrobial lock technic (ALT) on mixed biofilms non albicans Candida species -bacteria formed on surgical catheters, using ethylene diamine tetra-acetic acid (EDTA) and amphotericin B (AmB) as antimicrobial agents. Mixed biofilms of non-albicans Candida species -bacteria, co-isolated from the same medical device, were grown on catheters and incubated with EDTA and AmB lock solution for 1, 3, 5 or 7days. Our results suggest AmB associated to EDTA may be useful ALT agents in the treatment of catheter related infections.