The environmental Escherichia albertii strain DM104, which cross-reacts serologically with Shigella dysenteriae was assessed for pathogenic properties, immunogenicity, and protective efficacy in different animal models to evaluate it as a vaccine candidate against S. dysenteriae, which causes the severe disease, shigellosis. The DM104 isolate was found to be non-invasive and did not produce any entero- or cyto-toxins. The strain also showed negative results in the mouse lethal activity assay. The non-pathogenic DM104 strain gave, however, a high protective efficacy as an ocularly administered vaccine in the guinea pig eye model against S. dysenteriae type 4 challenge. It also induced a high titer of serum IgG against S. dysenteriae type 4 whole cell lysate and lipopolysaccharide. Taken together, all these results indicate a good potential for the use of the DM104 as a live vaccine candidate against shigellosis.