Influence of Growth Rate and Nutrient Limitation on the Gross Cellular Composition of Pseudomonas aeruginosa and Its Resistance to 3- and 4-Chlorophenol

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Abstract
Concentrations of 3-chlorophenol and 4-chlorophenol below their minimal inhibitory concentrations were found to increase the permeability of Pseudomonas aeruginosa cells to protons. Levels of such activity were assessed in suspensions of cells prepared from chemostat-grown cultures, limited by either magnesium (Mg-lim) or glucose (G-lim), with the use of five growth rates. Drug concentrations required to produce the same levels of proton translocation varied with the growth rate and the nutrient limiting growth. Fast-growing cultures were more sensitive than slower-growing ones, and G-lim cells were generally more sensitive than Mg-lim ones. 3-Chlorophenol had greater activity than 4-chlorophenol at slow growth rates, but at faster rates of growth their activity was similar. Variation in these iso-effective concentrations for different cells probably reflected an alteration in the ease of drug penetration of the outer envelope. Uptake of the compounds by cells in suspension varied, drug-sensitive bacteria absorbing more than resistant ones. This variation in uptake persisted when bacteria were solvent-extracted to remove readily extractable lipids (REL). Since no significant alteration in cell size was observed among the growth conditions studied, variation in absorption probably resulted from an altered affinity of the cells to the drug, with little involvement of REL. Overall REL content did not alter significantly with growth rate or nutrient limitation. However, total phospholipid content decreased and fatty acid content increased with increasing growth rate. For G-lim and Mg-lim cultures phosphatidylcholine content remained constant, yet phosphatidylethanolamine and phosphatidylglycerol content decreased with increasing growth rate. Diphosphatidylglycerol content decreased with increasing growth rate for Mg-lim cultures and remained relatively constant for G-lim cultures. Lipopolysaccharide content of the cells was higher in Mg-lim than in G-lim cultures and decreased with increasing growth rate in both cases. Lipopolysaccharide content correlated significantly with drug uptake and sensitivity, and it appeared to determine the degree of penetration of the cell envelope by these chlorinated phenols.