A facile one-pot method to prepare water-soluble, uniform-sized ZnS nanospheres (NSs) is reported. In the reaction, gradually decomposed thioacetamide (TAA) was used as the sulfur precursor, and cationic surfactant didodecyldimethylammonium bromide (DDAB) embedded with ferrocene (Fc) as the ligands for anions. The synthesized ZnS NSs were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy dispersive spectrometry (EDS), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). Wonderfully, the obtained ZnS NSs exhibited high performance in prohibiting the growth of Escherichia coli strain MG1655 (E. coli) and negligible mammalian cell toxicity, and these important features make it a promising antimicrobial agent for controlling implant-related infections.