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Journal of Antimicrobial Chemotherapy (2003) 51, 565-574
© 2003 The British Society for Antimicrobial Chemotherapy

Relationship between ß-lactamase production, outer membrane protein and penicillin-binding protein profiles on the activity of carbapenems against clinical isolates of Acinetobacter baumannii

Felipe Fernández-Cuenca1,*, Luis Martínez-Martínez1,2, Mª Carmen Conejo1, Juan A. Ayala3, Evelio J. Perea1,2 and Alvaro Pascual1,2

1 Department of Microbiology, University of Seville, Apdo. 914, 41009 Seville; 2 University Hospital Virgen Macarena, Seville; 3 Center of Molecular Biology ‘Severo Ochoa’, Autonomous University of Madrid, Madrid, Spain

Received 30 July 2002, returned 25 September 2002, revised 14 November 2002; accepted 20 November 2002

Twenty blood isolates of Acinetobacter baumannii were studied, representing eight pulsed-field gel electrophoresis patterns and all different antimicrobial susceptibility patterns observed during 1995–97 at the University Hospital Virgen Macarena, Seville, Spain. The MIC90s (mg/L) of imipenem and meropenem decreased from 16 to 0.5 and from 8 to 4, respectively, in the presence of BRL 42715 (BRL) but not clavulanic acid. Hydrolysing activity (nmol/min/mg) of bacterial supernatants against cefaloridine ranged from 8.8 to 552.3 for A. baumannii type I (imipenem MICs <= 2), which expressed only a ß-lactamase of pI >= 9, and from 12.3 to 1543.5 for A. baumannii type II (imipenem MICs >= 4), which expressed a ß-lactamase of pI >= 9 and two others of pI 6.3 and 7. The hydrolysing activities of A. baumannii type II against imipenem, meropenem and oxacillin were higher than those observed for A. baumannii type I. Ten outer membrane protein (OMP) profiles (A. baumannii types I and II) were visualized on 10% SDS–PAGE gels with 6 M urea, whereas only five OMP profiles (A. baumannii types I and II) were differentiated in 12% SDS–PAGE gels. Five A. baumannii with OMP profile type B, characterized by the absence of a 22.5 kDa OMP, were resistant to meropenem and/or imipenem. Twelve penicillin-binding protein (PBP) patterns were observed. PBP patterns of A. baumannii type II were characterized by the absence of a 73.2 kDa band (PBP 2). We concluded that production of ß-lactamases of pI 6.3 and 7.0 and reduced expression of PBP 2 are the most frequently observed mechanisms of resistance to carbapenems. In some isolates, loss of a 22.5 kDa OMP is also related to resistance to carbapenems.

Keywords: Acinetobacter baumannii, carbapenems, ß-lactamases, outer membrane proteins, penicillin-binding proteins

* Corresponding author. Tel: +34-95-455-2862; Fax: +34-95-437-7413; E-mail: felipefc{at}supercable.es


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