Silver and hydrogen peroxide (HP) acted synergistically on the viability of E. coli K-12. In certain concentration ranges the synergistic effect amounted to about 3 logs. Toxicity process kinetics were determined by following the decrease in luminescence of a highly luminescent recombinant E. coli harbouring a plasmid carrying the whole lux system of Vibrio fischeri. As in the viability studies, silver and HP also showed a synergistic effect on in vivo luminescence, which amounted to a 2 log increase in toxicity. A similar phenomenon was found for silver and certain metal ions, including Cu2+, Ni2+, Zn2+ and Cd2+, where toxicity increased by a factor of 10 in the presence of sub-inhibitory concentrations of silver. To monitor the expression of different stress response systems in treated cells, we have used E. coli carrying fusions of the lux system to promoters of different stress-controlling genes. Of the fusions tested, HP substantially increased the activity of recA, katG, micF, grpE, lon, and dnaK. Silver exerted a mild effect acting only on grpE and lon promoters. When in combination, a synergistic induction of the dnaK fusion and a slightly additive effect on grpE fusion were recorded. It appears that the combined toxic effect of silver and HP may be related with damage to cellular proteins. Nevertheless, the involvement of other cellular moieties can not be ruled out. The possibility that the synergistic effect is related to chemical interactions between silver and HP and the consequent increase in their toxicity is discussed.

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