The pathogenesis of both superficial and systemic candidiasis is closely dictated by properties of the yeast biofilms. Despite extensive investigations on bacterial biofilms, the characteristics of candidal biofilms, and various factors affecting this process remain to be determined. Therefore we examined the effect of human whole saliva and dietary sugars, glucose and galactose on the adhesion and biofilm formation of Candida albicans. Biofilms of C. albicans isolate 192 887 g were developed on polystyrene, flat-bottomed 96-well microtiter plates and monitored using ATP bioluminescence and tetrazolium (XTT) reduction assays as well as the conventional colony forming unit (CFU) evaluation. Our data showed that both the ATP and the XTT assays strongly correlated with the CFU assay (ATP versus CFU: r = 0.994, P = 0.006; XTT versus CFU: r = 0.985, P = 0.015). Compared with a glucose-supplemented (100 mM) medium, galactose containing (500 mM) medium generated consistently lower levels of both candidal adhesion and biofilm formation (all P < 0.05), but a higher pace of biofilm development over time (96 h). Whist the presence of an immobilised saliva coating had little effect on either the candidal adhesion or biofilm formation, the addition of saliva to the incubation medium quantitatively affected biofilm formation especially on day 3 and 4, without any significant effect on yeast adhesion. To conclude, biofilm formation of C. albicans within the oral milieu appears to be modulated to varying extents by dietary and salivary factors and, further investigations are required to elucidate these complex interactions.