Biofilms on silicone rubber voice prostheses are the major cause for frequent failure and replacement of these devices. The presence of both bacterial and yeast strains has been suggested to be crucial for the development of voice prosthetic biofilms. Polymicrobial biofilms that include Candida albicans and Rothia dentocariosa are the leading cause of voice prosthesis failure. An in vitro biofilm comprising these two organisms was developed on silicone rubber, a material used for Groningen button voice prosthesis. We found that this biofilm environment was not conducive for C. albicans growth or differentiation. Global transcriptional analyses of C. albicans biofilm cells grown with R. dentocariosa revealed that genes with functions related to cell cycle progression and hyphal development were repressed >2-fold. The mixed species biofilms were more compact and less robust compared to C. albicans mono-species biofilms, even when developed under conditions of continuous nutrient flow. Under these conditions R. dentocariosa also significantly inhibited C. albicans biofilm dispersal. Preferential adherence of R. dentocariosa to candidal hyphae was mediated by the adhesin Als3.