Dry matter was determined by drying samples at 65 °C
for 48 h to constant weight. Crude protein was analyzed by
the Kjeltec System (Tecator, Denmark) using AOAC methods
(1990). Neutral detergent Wber (NDF) and acid detergent
Wber (ADF) were determined according to the method
of Van Soest et al. (1991). pH was measured using a pH
meter (HI9321, Hanna Instrument, Portugal). Water soluble
carbohydrate was determined by the method
were achieved using 9 ml aliquots of sterile saline. The total
bacterial count (aerobes and facultative anaerobes) was
determined on plate count agar incubated at 30 °C for 48 h.
LAB were determined on MRS agar incubated at 37 °C for
24 h. Yeasts were determined on yeast-malt extract agar
(Difco Laboratories, USA) incubated at 37 °C for 48 h. Colony
counts of yeast and Bacillus spp. were determined by
selective counting characteristic colonies on the agar plates.