The fermentative assay was performed in triplicate at 23 °C using a
fresh free-run juice (no skin content) made from Syrah grapes (Vitis
vinifera L.) with an initial sugar content of 220 g/l (potential alcohol
content of 13% v/v), pH 3.5, and heat treated at 100 °C for 3 min. All inocula
were standardized in order to obtain homogenous active populations
(106 cfu/ml), by adding 100 μl of each strain to 5 ml of YEPD
medium (Kurtzman & Fell, 1998) and were grown for 24 h at 23 °C,
twice in succession. In sequential fermentations (SF), 70 ml of must in
100 ml flasks was inoculated with 1 ml of each non-Saccharomyces
strain, and after 7 days (when a level of 8% v/v ethanol was reached)
the second inoculation was performed with 1 ml of strain Sc7VA. On
the other hand, mixed fermentations (MF) were co-inoculated with
1 ml of a non-Saccharomyces strain and 100 μl of strain Sc7VA (non-
Saccharomyces:Saccharomyces ratio 10:1).
Fermentation kinetics and fermentative power were estimated by
the dailyweighing of the fermentation flasks, thus registering variations
that correspond to the loss of CO2 associated with the fermentative
process. All chemical analyses were performed at the end of the
fermentations.