icrobiological analysis (total viable bacterial count)
Total viable counts were determined during the fermentation.
The fermented soybeans (10 g) were homogenized with 90 ml of
the sterilized physiological saline (0.85%). Serial dilutions were
prepared in sterilized physiological saline and 1 ml of appropriate
dilutions was poured in triplicate plates for total viable count. Total
viable counts of L. plantarum P-8, L. casei Zhang and B. animalis 937
were made using a pour plate method and MRS agar after serial
dilution in maximum recovery diluents. A pre-prepared test sample
(1 ml) of 107, 108, and/or 109 dilution was transferred into a
sterile petri dish, in triplicate, and warm (45 2 C) sterile plate
count MRS agar (15 ml) was mixed with the inoculums. Cultures
were incubated anaerobically at 37 C for 48 2 h. Total Viable
counts of B. subtilis natto was made using spread plate technique.
About 15 ml nutrient agar medium was poured into plates prior to
use. A pre-prepared test sample (0.1 ml) of 106, 107, and/or 108
dilutionwas transferred to the surface of nutrient agar medium, the
inoculums was spread evenly over the entire surface of the agar by
a rotary twirling motion of the plate under the rod. Cultures were
incubated at 37 C for 24 2 h. The colonies were then counted and
expressed as logarithmic colony forming units per gram (lg CFU/g)
of the sample.