Role of beta-lactamase in in vivo development of ceftazidime resistance in experimental Pseudomonas aeruginosa endocarditis.
Two ceftazidime-resistant variants of Pseudomonas aeruginosa (PA-48, PA-60), obtained from cardiac vegetations of rabbits with endocarditis receiving ceftazidime therapy, were studied for mechanisms of resistance. Both resistant variants were stably derepressed for the type Id beta-lactamase, which was ceftazidime inducible in the parental strain (PA-96) used to initially infect the rabbits. There was no evidence of ceftazidime bioinactivation by the resistant strains, and their outer membrane permeabilities were comparable to those of the parental strain. No alterations were observed in patterns of outer membrane proteins or membrane lipopolysaccharides in the resistant variants as compared with the parental strain. Penicillin-binding protein patterns of the resistant variants revealed the absence of penicillin-binding protein 4 in both, with acquisition of a new protein of higher apparent molecular weight in PA-60. Calculation of the rate of appearance of ceftazidime in the periplasm at sub-MICs suggested that slow enzymatic hydrolysis of the beta-lactam, rather than nonhydrolytic trapping, was the major explanation for the induced resistance in vivo in strains PA-48 and PA-60.
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis; D001426:Bacterial Proteins; D002352:Carrier Proteins / Q000032:analysis; D002442:Ceftazidime / Q000378:metabolism / Q000494:pharmacology* / Q000627:therapeutic use; D002463:Cell Membrane Permeability / Q000187:drug effects; D004352:Drug Resistance, Microbial; D004591:Electrophoresis, Polyacrylamide Gel; D004697:Endocarditis, Bacterial / Q000188:drug therapy / Q000382:microbiology*; D006602:Hexosyltransferases; D008070:Lipopolysaccharides / Q000032:analysis; D002267:Muramoylpentapeptide Carboxypeptidase / Q000032:analysis; D046915:Penicillin-Binding Proteins; D010458:Peptidyl Transferases; D011552:Pseudomonas Infections / Q000188:drug therapy / Q000382:microbiology*; D011550:Pseudomonas aeruginosa / Q000032:analysis / Q000187:drug effects* / Q000201:enzymology; D011817:Rabbits; D001618:beta-Lactamases / Q000378:metabolism*
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D008070:Lipopolysaccharides; D046915:Penicillin-Binding Proteins; D002442:Ceftazidime; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D002267:Muramoylpentapeptide Carboxypeptidase; D001618:beta-Lactamases