In this study, the safety evaluation, health-beneficial potential, probiotic and functional properties of Lactobacillus delbrueckii MA-9 strain originating from human breast milk were investigated in vitro. MA-9 strain showed very high resistance to simulated in vitro gastrointestinal conditions and food preservatives. The strain demonstrated non-hemolytic activity, inhibitory action on different indicator pathogen microorganisms (twelve ­clinical or human food-borne bacteria, two yeast and seven bacterial fish pathogens), high levels of aggregation, strong cholesterol removal ability and 2,2-Diphenyl-1-­picrylhydrazyl (DPPH) radical scavenging activity (43.40%). Moreover, the strain showed positive a-amylase and lichenase (ß-1,3-1,4-glucanase) enzyme activities. The results demonstrated that the L. delbrueckii MA-9 strain can be a good probiotic candidate and natural food additive to promote health.