Biofilms of Staphylococcus epidermidis and Candida spp. are two of the most frequent factors of infections associated with the use of indwelling medical devices. Several strategies have been proposed and/or developed to prevent infection. The aim of this study was to compare the effect of sub-inhibitory concentrations of anti-microbial agents on biofilm formation. Biofilms of three strains of S. epidermidis and two of both Candida albicans and Candida dubliniensis were formed in the presence of three antibiotics and two antifungal agents respectively. Based on the control samples, the percentage of biofilm formation inhibition by the different agents was determined and compared. The results showed that the influence of the antibacterial and antifungal agents tested is strain dependent, with the effect of the different agents also varying among strains, even though they have the same mechanism of action.