Fluconazole resistance of Candida albicans (C. albicans) is a growing medical issue which makes patient management more difficult. The development of new effective, safe and inexpensive alternatives becomes essential. Probiotics represent an intriguing strategy that can be used for prevention or treatment of Candida infections. The aim of our study is to investigate the anti-fungal activity of various Lactobacillus species against fluconazole resistant C. albicans isolates and to assess the inhibitory effect of cell free supernatant of most potent one. Sixty-nine C. albicans isolates were obtained from different clinical specimens and subjected to antifungal susceptibility testing. Then, the anti-Candida activity of various Lactobacillus spp was investigated via different methods. Results showed that 33% of C. albicans isolates were fluconazole resistant with MIC ≥ 64 μg/ml. L. rhamnosus demonstrated strong inhibition activity against majority of fluconazole resistant C. albicans isolates, followed by L. reuteri and L. salivarus respectively. Inhibitory activity of L. rhamnosus against C. albicans may be due to acid production. Our findings conclude that L. rhamnosus represent a promising candidate to be used in the treatment of candidiasis caused by fluconazole-resistant C. albicans strains.