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Antimicrobial Agents and Chemotherapy, May 2003, p. 1536-1542, Vol. 47, No. 5
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.5.1536-1542.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Chromosomal Integration of a Cephalosporinase Gene from Acinetobacter baumannii into Oligella urethralis as a Source of Acquired Resistance to ß-Lactams

Hedi Mammeri,1,2 Laurent Poirel,1 Nicole Mangeney,2 and Patrice Nordmann1*

Service de Bactériologie-Virologie, Hôpital de Bicêtre, Assistance Publique/Hôpitaux de Paris, Faculté de Médecine Paris-Sud, 94275 Le Kremlin-Bicêtre,1 Laboratoire Central, Hôpital Albert Chenevier, 94000 Créteil, France2

Received 25 November 2002/ Returned for modification 7 January 2003/ Accepted 29 January 2003

Clinical Oligella urethralis isolate COH-1, which was uncommonly resistant to penicillins and narrow-spectrum cephalosporins, was recovered from a 55-year-old patient with a urinary tract infection. Shotgun cloning into Escherichia coli and expression experiments gave recombinant clones expressing either an AmpC ß-lactamase-type phenotype of resistance or a carbenicillin-hydrolyzing ß-lactamase-type phenotype of resistance. The AmpC ß-lactamase identified (ABA-1), which had a pI value of 8.2, had 98% amino acid identity with a chromosomally encoded cephalosporinase of Acinetobacter baumannii. A 820-bp insertion sequence element, ISOur1, belonging to the IS6 family of insertion sequence elements, was identified immediately upstream of blaABA-1, providing a -35 promoter sequence and likely giving rise to a hybrid promoter region. The carbenicillin-hydrolyzing ß-lactamase identified (CARB-8), which had a pI value of 6.4, differed from CARB-5 by two amino acid substitutions. Hybridization of CeuI fragment I-restricted DNA fragments of O. urethralis COH-1 with blaABA-1-, blaCARB-8-, and 16S rRNA-specific probes indicated the chromosomal integration of the ß-lactamase genes. PCR and hybridization experiments failed to detect blaCARB-8- and blaABA-1-like genes in three O. urethralis reference strains, indicating that the ß-lactamase genes identified were the source of acquired resistance in O. urethralis COH-1. This is one of the few examples of the interspecies transfer and the chromosomal integration of a gene encoding a naturally occurring ß-lactamase.

* Corresponding author. Mailing address: Service de Bactériologie-Virologie, Hôpital de Bicêtre, 78 rue du Général Leclerc, 94275 Le Kremlin-Bicêtre Cedex, France. Phone: 33-1-45-21-36-32. Fax: 33-1-45-21-63-40. E-mail: nordmann.patrice{at}bct.ap-hop-paris.fr.

Antimicrobial Agents and Chemotherapy, May 2003, p. 1536-1542, Vol. 47, No. 5
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.5.1536-1542.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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