We report on the preparation and characterization of electrospun polyurethane nanofibers containing silver, cactus, rosin and Scutellariae Radix. The utilized polyurethane nanofibers containing different composite materials were prepared by a simple dip coating method. The morphology, structure and thermal characteristics of as-prepared composite nanofibers were studied by scanning electron microscopy, X-ray diffraction, Fourier transform infrared, Raman spectroscopy and thermogravimetric analysis. The antimicrobial activity of the composite nanofibers was tested against two common food borne pathogenic bacteria, Staphylococcus aureus and Escherichia coli, by the minimum inhibitory concentration method. Our results demonstrated that more pronounced antimicrobial activities were observed for the composite nanofibers. Overall, the fabrication of cheap, stable and effective material with excellent antimicrobial activity can be utilized to inhibit the microbial growth associated with food stuff.