The article presents the incorporation of biocides [2-substituted N-alkylimidazoles and their silver(I) complexes] into electrospun nylon 6 nanofibers for application as antimicrobial materials. The electrospun nylon 6/biocides nanofiber composites were characterized by IR spectroscopy (ATR-FTIR) and scanning electron microscopy (SEM-EDX). The antimicrobial activity of the electrospun nylon 6/biocides nanofiber composites was evaluated against Escherichia coli, Staphylococcus aureus, and Bacillus subtilis subsp. spizizenii using the disk diffusion method, the American Association for Textile Chemists and Colorists test method 100-2004 and the dynamic shake flask method (American Society for Testing and Materials E2149-10). The electrospun nylon 6 nanofibers incorporated with 2-substituted N-alkylimidazoles displayed moderate to excellent levels of growth reduction against S. aureus (73.2–99.8%). For the electrospun nylon 6 nanofibers incorporated with silver(I) complexes, the levels of growth reduction were >99.99%, for both E. coli and S. aureus, after the antimicrobial activity evaluation using the shake flask method. The study demonstrated that the electrospun nanofibers, fabricated using the incorporation strategy, have the potential to be used as attractive antimicrobial materials. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014, 131, 39783.