Silver nanoparticle-embedded polyrhodanine nanofibers were synthesized by chemical oxidation polymerization. Silver ions were reduced to silver nanoparticles by oxidizing rhodanine monomer and simultaneously complexed with the rhodanine due to coordinative interactions, resulting in the formation of silver nanoparticle-embedded polyrhodanine nanofibers. The synthesized silver/polyrhodanine nanofiber was found to have the excellent antimicrobial activity against Gram-negative Escherichia coli, Gram-positive Staphylococcus aureus, and Candida albicans. The modified Kirby-Bauer test demonstrated that the silver/polyrhodanine nanofiber had better antimicrobial efficacy than silver sulfadiazine.