ABSTRACT The effects of ethylenediaminetetraacetic acid (EDTA) and its salts, alone or in combination with Mg 2+ , Fe 3+ , Zn 2+ or Ca 2+ , on growth of S. aureus strain 196E, were studied. EDTA or its Na + or K + salts (0.8–1.7 mM), but not its Ca 2+ or Fe 3+ salts, inhibited cell growth in Brain Heart Infusion broth. Bacteriostatic effects in Casamino Acids‐Yeast Extract (CasY) broth were produced by 0.17 mM EDTA. Addition of Fe 3+ , Zn 2+ , or Ca 2+ to EDTA‐containing broth eliminated inhibition at an EDTA: cation molar ratio of 1:1, while Mg 2+ required a sevenfold concentration. Data suggest that EDTA exerts its inhibitory effect by chelating calcium and/or other essential cations which form complexes with comparable stability constants with EDTA.