Antimicrobial activity of three kinds of commercially available montmorillonite nano-clays including a naturally occurring one (Cloisite Na+) and two organically modified ones (Cloisite 20A and Cloisite 30B) against four representative pathogenic bacteria (two Gram-positive ones such as Staphylococcus aureus and Listeria monocytogenes, and two Gram-negative ones such as Salmonella typhimurium and E. coli O157:H7) was investigated. Antimicrobial activity was found to be dependent on the type of nano-clay and microorganisms tested. Among the nano-clays tested, Cloisite 30B showed the highest antibacterial activity followed by Cloisite 20A, however, the unmodified montmorillonite (Cloisite Na+) did not show any antibacterial activity. Especially, Cloisite 30B inactivated Gram-positive bacteria completely within an hour of incubation and inactivated Gram-negative bacteria by more than 2-3 log cycles after 8 hours incubation. SEM and TEM images of cell structure indicated that the organically modified nano-clay caused rupture of cell membrane and inactivation of the bacteria. This finding of Antimicrobial activity of the organo-clay would open a new opportunity to develop polymer nanocomposites with additional functionality, i.e., Antimicrobial function.