We investigated the effects of cations on the stability in culture of mucoid strains pf Pseudomonas aeruginosa isolated from patients with cystic fibrosis by studying their effects on the selection of nonmucoid derivatives which arise by spontaneous mutation in cultures of mucoid organisms. Calcium ion concentrations in the range 0.55 to 1.85 mM had no effect on the growth or stability of the mucoid cultures. Higher levels (5.0 mM) inhibited the growth of both mucoid and nonmucoid cells. Likewise, magnesium ion in concentrations of 0.3 to 3.0 mM had no effect. The concentration of iron (either Fe2+ or Fe3+) had a profound effect on the selection of nonmucoid mutants in unshaken cultures of mucoid organisms. In medium containing 0.01 mM iron, nonmucoid mutants rapidly accumulated to a greater than 100-fold-higher frequency than the mucoid forms. Rates of accumulation of nonmucoid derivatives were lower in media containing lower concentrations of iron. The possible role of iron in the selection of nonmucoid cells from a population of mucoid P. aeruginosa is discussed.