Copper Acetate Manufacturers (WSDTY) proposed we hypothesized that one mechanism underlying this inhibitory synergistic effect might be reduction of Copper Acetate by Fe(II). Fe(II) has been shown to reduce Copper Acetate under environmentally relevant conditions (35). Our own experiments showed that in our growth medium, Fe(II) could quickly reduce a significant amount of Cu(II) to Cu(I) abiotically. We tested copper reduction as a possible cause of the inhibitory effect by using ascorbate as an alternate reductant to Fe(II): when added at the same concentrations as Fe(II), ascorbate also rapidly reduced Cu(II) but resulted in only a slight delay in growth compared to that with Fe(II). Moreover, Cu(I) had the same effect on R. palustris as Cu(II) either alone, with ascorbate, or with Fe(II). If the reduction of Cu(II) by Fe(II) were the sole cause of toxicity, then we would expect Cu(I) to have a greater effect than Cu(II). Together, these results suggest that the reduction of Cu(II) to Cu(I) is not sufficient to explain the observed growth inhibition.
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