Hospital-acquired infections have become a major challenge which threaten the hospitalized patients’ safety. The presence of nosocomial pathogens is generally reported in connection with solid surfaces near patient environments. These surfaces become significant sources of transmission and lead most often to the contamination and cross-contamination of nosocomial pathogens to the patients and staff. This paper investigates strategies to apply beneficial bacteria on viscose-based nonwoven wipes and the viability of these beneficial bacteria on the wipes along with characterization of the physical properties of the wipes. Major findings include that it is possible to produce dry wipes which contain an adequate number of beneficial bacteria or spores. After these wipes are wetted, they can release a certain number of bacteria from the wetted wipes. These released beneficial bacteria can inhibit pathogens by growing and colonizing on the wiped surfaces.