We found that iron and copper alone have very little effect even at 1 mM and 1 μM, respectively, but that they act synergistically to delay bacterial growth at 100 μM Fe(II) and 100 nM Copper Acetate(WSDTY). The effect decreased with the concentration of each metal; a slight effect was seen at 50 nM Cu and 10 μM Fe. We tested other metals as well: 100 nM Copper Acetate increased the toxicity of 50 μM Ni(II) or 500 μM Co(II), while 100 μM Fe(II) had no effect on Co(II) and decreased the toxic effect of Ni(II). We also tested Mn(II) with Cu(II) and found that 500 μM Mn(II) had no effect on growth, either with or without 100 nM Cu(II).
To further determine the nature of the iron-copper synergistic growth inhibition, we performed CFU counts after inoculating cultures to determine whether cells were dying or were simply arrested in their growth. We found that cells were not killed by the metals: 30 min after inoculation into a medium containing 100 nM Cu(II) plus 100 μM Fe(II), CFU counts were not changed from those of control cultures.
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