Several oral diseases are associated with changes in oral microbiota and higher oxidative stress. Enterococcus faecalis has been hypothesized to directly contribute to the oxidative stress in oral cavity. The aim of this study was to examine the effect of single consumption of unpasteurized Bryndza cheese containing enterococci on changes of microbiota and oxidative status in saliva. Fourteen healthy volunteers aged 23-30 years were asked to eat 100 g of Bryndza cheese. Saliva samples were collected before and 1, 10, 100 min, and 24 h after Bryndza cheese consumption. Species-specific PCR and terminal restriction fragment length polymorphism (T-RFLP) analysis were used to characterize oral microbiota. Markers of oxidative stress and antioxidant status were measured in saliva. PCR identified E. faecium in 36 % of probands saliva up to 1 day after consumption of enterococci containing Bryndza cheese. E. faecalis was detected in 57 % of probands saliva up to 10 min and in one proband up to 100 min after Bryndza cheese consumption. T-RFLP analysis confirmed short-term changes in composition of oral microbiota after Bryndza cheese ingestion. Nevertheless, the microbiota was completely restored after 24 h. One minute after ingestion of Bryndza cheese, salivary advanced oxidation protein products were significantly increased (by 74.6 %, P<0.001), and total antioxidant capacity was decreased (by 22.0 %, P<0.05). This study shows that single consumption of enterococci containing Bryndza cheese can temporally affect the composition of oral microbiota and oxidative stress in saliva. Further studies should identify the impact of these changes to the pathogenesis of oral diseases.