Novel hybrid polyvinyl butyral nanofibers have been developed for antimicrobial applications. The nanofiber mats were obtained from a needleless rod electrospinning system. The novel inorganic antibacterial agents were incorporated into the nanofibers, and their antibacterial activity was compared. The obtained nanoparticle/nanofiber hybrid mats have a good surface morphology. The results indicated that the CuO, ZnO, ZnO/TiO2, and AgNO3 nanoparticle‐incorporated nanofiber layers have excellent antibacterial activity against to Escherichia coli compared with TiO2, SnO2, and ZrO2 ones. Copyright © 2016 John Wiley & Sons, Ltd.