Control measures against antimicrobial resistant bacterial pathogens are important challenges in our daily life. In this study, we discuss the sensitivity and resistance of four bacterial pathogens, Vibrio alginolyticus, Escherichia coli, Staphylococcus aureus, and Bacillus subtilis, to silver-silica hybrid nanoparticles. Successively, by combining with an efflux pump blocking agent Verapamil, we find that these hybrid nanoparticles induce complete mortality to even the most resistive S. aureus. The above pathogens are selected from a pool of 100 bacterial strains resistant to silver nitrate. While S. aureus shows increased resistance to the nanoparticles, the cell wall integrity and genetic stability of V. alginolyticus and E. coli are compromised in the presence of the hybrid nanoparticles. These studies suggest that the antimicrobial properties of the nanoparticles against Gram-negative pathogens originate from increased oxidative stress, which is confirmed by the blocking of reactive oxygen species (ROS) using scavengers such as ascorbic acid and observing DNA damage. The antimicrobial property of the nanoparticle when combined with its nontoxic nature to mammalian cells makes it a promising agent for controlling drug-resistant Gram-negative pathogens. © 2019 American Chemical Society.