Lyophilized culture of L. acidophilus La5 was rehydrated (at 15 C
for 20 min) in sterile reconstituted skimmed milk containing 20%
(v/v) glycerol (cryoprotectant), and transferred to micro tubes and
kept in a freezer (18 C). For processing, this culture was reactivated
in MRS broth (2% v/v) for two consecutive times, with incubation
at 37 C for 15 h. The reactivated culture was placed in
sterile polypropylene tubes and centrifuged at 8000 g for 10 min at
4 C (Sorvall Instruments RC5C e DuPont). The cell concentratewas
resuspended in peptone water (0.1% v/v) and centrifuged again
under the same conditions, followed by resuspension in peptone
water to obtain a concentration of cells with 9e10 log10 CFU/mL
(Gebara et al., 2013). The cell concentrate viability was assessed by
pour plating on MRS agar incubated at 37 C for 72 h in anaerobic