title: Roles of beta-lactamases and porins in activities of carbapenems and cephalosporins against Klebsiella pneumoniae.
abstract: Two clinical isolates of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae were noted to be less susceptible than expected to imipenem. Both were missing outer membrane proteins that serve as channels for antibiotic entry. The role of beta-lactamase in resistance was investigated by eliminating the original ESBL and introducing plasmids encoding various ESBLs and AmpC beta-lactamase types, by studying the effect of an increased inoculum, and by evaluating interactions with beta-lactamase inhibitors. The contribution of porin deficiency was investigated by restoring a functional ompK36 gene on a plasmid. Plasmids encoding AmpC-type beta-lactamases provided resistance to imipenem (up to 64 microg/ml) and meropenem (up to 16 microg/ml) in strains deficient in porins. Carbapenem resistance showed little inoculum effect, was not affected by clavulanate but was blocked by BRL 42715, and was diminished if OmpK36 porin was restored. Plasmids encoding TEM- and SHV-type ESBLs conferred resistance to cefepime and cefpirome, as well as to earlier oxyimino-beta-lactams. This resistance was magnified with an increased inoculum, was blocked by clavulanate, and was also lowered by OmpK36 porin restoration. In addition, SHV-2 beta-lactamase had a small effect on carbapenem resistance (imipenem MIC, 4 microg/ml, increasing to 16 microg/ml with a higher inoculum) when porins were absent. In K. pneumoniae porin loss can thus augment resistance provided either by TEM- or SHV-type ESBLs or by plasmid-mediated AmpC enzymes to include the latest oxyimino-beta-lactams and carbapenems.
authors.affiliation: Department of Microbiology, School of Medicine, University of Seville, Seville, Spain. lmartin@cica.es
authors.forename: L
authors.initials: L
authors.lastname: Martínez-Martínez
authors.forename: A
authors.initials: A
authors.lastname: Pascual
authors.forename: S
authors.initials: S
authors.lastname: Hernández-Allés
authors.forename: D
authors.initials: D
authors.lastname: Alvarez-Díaz
authors.forename: A I
authors.initials: AI
authors.lastname: Suárez
authors.forename: J
authors.initials: J
authors.lastname: Tran
authors.forename: V J
authors.initials: VJ
authors.lastname: Benedí
authors.forename: G A
authors.initials: GA
authors.lastname: Jacoby
mesh_terms: D015780:Carbapenems / Q000494:pharmacology*; D002511:Cephalosporins / Q000494:pharmacology*; D004352:Drug Resistance, Microbial; D007711:Klebsiella pneumoniae / Q000187:drug effects*; D008826:Microbial Sensitivity Tests; D018272:Porins / Q000502:physiology*; D001618:beta-Lactamases / Q000502:physiology*
chemical_list: D015780:Carbapenems; D002511:Cephalosporins; D018272:Porins; D001618:beta-Lactamases