Inocula preparetion of yeasts and E. betae
Pichia albicans and C. sake were grown at 24°C for 48 h on nutrient
yeast broth (20.0 g glucose; 20.0 g peptone and 10.0 g yeast extract) with
shaking. Cultures were centrifuged and then pellets were resuspended in sterile
distilled water and centrifuged again. The resulting pellets were dispersed in
sterile distilled water, and the concentration of the yeasts was adjusted to 108
colony forming units (cfu) ml-1 using a hemacytometer. 100 g of S. servisiae as
fresh preparations from dry commercial formula was well dissolved in sterile
distilled water and the suspension was diluted to 10 liters using sterile distilled
water before application.