Among Bacillus bacteria, B. subtilis is the species that produces the most antimicrobial compounds. In this study, we analyzed the activity of probiotic strain B. subtilis 3 against the influenza virus. The antiviral effect of this strain has been demonstrated in vitro and in vivo. A new peptide, P18, produced by the probiotic strain was isolated, purified, chemically synthesized, and characterized. Cytotoxicity studies demonstrated no toxic effect of P18 on Madin-Darby canine kidney (MDCK) cells, even at the highest concentration tested (100 g/ml). Complete inhibition of the influenza virus in vitro was observed at concentrations of 12.5 to 100 g/ ml. The protective effect of P18 in mice was comparable to that of oseltamivir phosphate (Tamiflu). Further study will assess the potential of peptide P18 as an antiviral compound and as a promising candidate for the development of new antiviral vaccines.