Denture stomatitis (DS) can affect about 70% of denture wearers causing inflammation of oral mucosa underneath the removable denture mostly seen at the palatal area. Candida albicans is a well-known as the main causative agent associated with denture stomatitis. Studies shown that DS-related biofilm consists of complex structured of microbial communities. Presence of co-existing bacteria in these biofilms may also contribute to the infection. In order to understand the microbial interaction between C. albicans and bacteria, this study focus on the ability of bacteria species which are Streptococcus mutans and Staphylococcus aureus to form biofilm with and without the presence of C. albicans. For this study, two groups were observed which are the single species biofilm and the mixed species biofilm. Biofilm assay was utilized to measure biofilm mass using spectrophotometer. From the observation, single species groups formed a better biofilm when compared to fungal-bacterial biofilm. Even though biofilm assay is unable to determine the number of cell available, both species were present when fungal-bacterial biofilm sample was observed microscopically using Gram stain technique. This experiment showed that both fungal and bacteria species are able to form biofilm together and this finding can be investigated further for better understanding of DS-related biofilm. KEYWORDS Biofilm, Candida albicans, mixed species biofilm