To demonstrate the feasibility of growing lactobacilli and producing lactic acid using maple sap as a sugar source and to show the importance of oligosaccharides in the processes.Two maple sap samples (Cetta and Pinnacle) and purified sucrose were used as carbon sources in the preparation of three culture media. Compared with the sucrose-based medium, both maple sap-based media produced increased viable counts in two strains out of five by a factor of four to seven. Maple sap-based media also enhanced lactic acid production in three strains. Cetta sap was found to be more efficient than Pinnacle sap in stimulating lactic acid production and, was also found to be richer in various oligosaccharides. The amendment of the Pinnacle-based medium with trisaccharides significantly stimulated Lactobacillus acidophilus AC-10 to grow and produce lactic acid.Maple sap, particularly if rich in oligosaccharides, represents a good carbon source for the growth of lactobacilli and the production of lactic acid.This study provides a proof-of-concept, using maple sap as a substrate for lactic acid production and for the development of a nondairy probiotic drink.