The advent of omic platforms revealed the significant benefits of probiotics in the prevention of many infectious diseases. This led to a growing interest in novel strains of probiotics endowed with health characteristics related to microbiome and immune modulation. Therefore, autochthonous bacteria in plant ecosystems might offer a good source for novel next-generation probiotics. The main objective of this study was to analyze the effect ofRouxiella badensis acadiensis Canan(R. acadiensis)a bacterium isolated from the blueberry biota, on the mammalian intestinal ecosystem and its potential as a probiotic microorganism.R.acadiensis, reinforced the intestinal epithelial barrier avoiding bacterial translocation from the gut to deep tissues, even after feeding BALB/c mice for a prolonged period of time. Moreover, diet supplementation withR. acadiensisled to increases in the number of Paneth cells, well as an increase in the antimicrobial peptide α defensin. The anti-bacterial effect ofR.acadiensisagainstStaphylococcus aureusandSalmonella entericaserovar Typhimurium was also reported. Importantly,R. acadiensis-fed animals showed better survival in an in vivoSalmonella entericaserovar Typhimurium challenge compared with those that received a conventional diet. These results demonstrated thatR. acadiensispossesses characteristics of a probiotic strain by contributing to the reinforcement and maintenance of intestinal homeostasis.