The raffinose family of oligosaccharides (RFOs) isolated from lupin seeds (Lupinus albus var. Multolupa) was evaluated for bifidogenic effects during the manufacture of probiotic fermented milk. A mixed starter inoculum was composed of Bifidobacterium lactis Bb-12 and Lactobacillus acidophilus (1:1). Lupins are a rich source of RFOs that can be used as functional food ingredients. The addition of RFOs to milk increased B. lactis Bb-12 and L. acidophilus populations at the final fermentation time compared with controls. Final fermentation products are positively affected by addition of RFOs, and time of fermentation was reduced from 12 to 10 h. When RFOs were added to milk, they were preferentially used as a carbon source (57.7%) compared with lactose (23.7%) at the end of fermentation. These results suggest that the eventual choice of B. lactis Bb-12 and L. acidophilus in a mixed culture at a 1:1 ratio and addition of RFOs to produce a fermented milk product would have the advantages of rapid growth and acidification rate and would likely increase the probiotic effect of the final functional product. Copyright ©, International Association for Food Protection.