Candida krusei and Candida albicans present the ability to form communities of microorganisms called biofilms. Biofilms can be composed of a single species or more and are an important virulence factor. The inhibition of C. albicans and C. krusei as well as of their dual-species biofilms by antimicrobial Photodynamic Therapy (aPDT) has been demonstrated. This study aimed to investigate the effect of aPDT, with TBO, on dual-species biofilms of C. albicans and C. krusei using different culture mediums, RPMI-1640 and Sabouraud-dextrose broth (SDB) to produce biofilms presenting different C. albicans/C. krusei ratio. Biofilms formed using RPMI-1640 presented a higher C. albicans/C. krusei ratio, however, biofilms formed using SDB presented a predominance of C. krusei. The metabolic activity of biofilms produced using RPMI-1640 was inhibited by aP (-40%), while biofilms produced using SDB were not affected by aPDT. In addition, biofilm biomass was reduced in biofilms produced using RPMI-1640 and treated with aPDT (-20%). The results demonstrated that aPDT reduces C. albicans development in dual-species biofilms with C. krusei. However, no effect could be observed on C. krusei, demonstrating that C. krusei, when present in the structure of dual-species biofilms can be resistant to aPDT.