Biofilms formed by pose therapeutic challenges due to their resistance to conventional antimicrobials, highlighting the need for more effective treatments. Rhamnolipids (RLs) are biosurfactants with diverse antimicrobial properties. Bacteriophages are viruses that target specific bacterial strains. Recent studies have shown that they may affect biofilm formation by fungi and yeasts. This study investigated the combined antimicrobial effects of RLs and bacteriophages against biofilms, focusing on their anti-adhesive and inhibitory effects on biofilm development. RT-PCR assays were used to analyze gene modulation in biofilm formation in response to RLs and bacteriophage treatments, while hyphae formation was examined using microscopy. The results showed that RLs-bacteriophage combinations significantly reduced biofilm formation compared to individual treatments. A combination of 200 mg/L RLs with bacteriophage BF9 led to a 94.8% reduction in biofilm formation. In a subsequent model, the same RL concentration with bacteriophage LO5/1f nearly eliminated biofilm formation (~96%). Gene expression analysis revealed downregulation of key biofilm-associated genes when cells were treated with 200 mg/L RLs and four bacteriophages (BF17, LO5/1f, JG004, FD). These results show the potential of RL and bacteriophage combinations in combating biofilms, presenting a promising therapeutic approach against resilient infections.