Transient loss of plasmid-mediated mercuric ion resistance after stress in Pseudomonas aeruginosa

Abstract

After freezing and thawing, Pseudomonas aeruginosa harboring a drug resistance plasmid (Hg2+r, Strr), became acutely sensitive to mercuric ions but not to streptomycin in the plating medium, whereas its sensitivity to both agents became more pronounced indicating a synergistic effect. This freeze-thaw-induced sensitivity was transient and capable of being repaired to a simple salts medium. Transient outer and cytoplasmic membrane damage was also observed in frozen and thawed preparations. From kinetics studies, repair of cytoplasmic membrane damage superseded repair of outer membrane damage and damage measured by mercuric ions and mercuric ions plus streptomycin. Osmotically shocked cells were also sensitive to mercuric ions, mercuric ions plus streptomycin, and sodium lauryl sulfate, but not to sodium chloride or streptomycin alone. This sensitivity was again transient and capable of repair in the same simple salts medium. Active transport of a non-metabolizable amino acid, alpha-amino isobutyric acid, was sensitive to mercuric ions and became more so after freezing and thawing. A freeze-thaw-resistant mercuric ion-dependent reduced nicotinamide adenine dinucleotide phosphate oxidoreductase was localized in the cytoplasm of this organism. This enzyme and an intact outer membrane appear to be required for mercuric ion resistance in this strain.