Tube dilution experiments showed that benzalkonium chloride (BC)-resistant mutants of Pseudomonas aeruginosa grown in the presence of 1,000 μg of BC per ml were at least 20 times more sensitive to polymyxin B and colistin sulfate than the BC-sensitive (BCS) parent strain. BCS cells selected for resistance to 500 μg of polymyxin B per ml remained sensitive to BC. There was little difference in the amount of carbenicillin, gentamicin sulfate, or rifampin needed to prevent growth of either the BCS or BC-resistant (BCR) strains. Growth of BCR cells was inhibited by ethylenediaminetetraacetate at a concentration of 400 μg/ml or less, whereas the BCS strain grew at ethylenediaminetetraacetate levels of 10,000 μg/ml. Phenylmercuric acetate and thimerosal inhibited growth of BCR and BCS cells at concentrations of 10 μg/ml or less. BCR cells were cross-resistant to >1,000 μg/ml concentrations of five other quaternary ammonium compounds, including three with C16 alkyls and two with alkyl groups of shorter length. The BCS strain was also resistant to >1,000 μg/ml concentrations of the three quaternary ammonium compounds with C16 alkyl groups but, in addition to BC, was inhibited by 200 μg/ml levels or less of the two quaternary ammonium compounds containing alkyl groups of less than 16 carbon atoms.