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Antimicrobial Agents and Chemotherapy, July 2002, p. 2162-2168, Vol. 46, No. 7
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.7.2162-2168.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

New Carbenicillin-Hydrolyzing ß-Lactamase (CARB-7) from Vibrio cholerae Non-O1, Non-O139 Strains Encoded by the VCR Region of the V. cholerae Genome{dagger}

Roberto Melano, Alejandro Petroni, Alicia Garutti, Héctor Alex Saka, Laura Mange, Fernando Pasterán, Melina Rapoport, Alicia Rossi,,{ddagger} and Marcelo Galas*

Servicio Antimicrobianos, Departamento de Bacteriología, Instituto Nacional de Enfermedades Infecciosas-ANLIS "Dr. Carlos G. Malbrán," Buenos Aires, Argentina

Received 24 August 2001/ Returned for modification 12 January 2002/ Accepted 7 April 2002

In a previous study, an analysis of 77 ampicillin-nonsusceptible (resistant plus intermediate categories) strains of Vibrio cholerae non-O1, non-O139, isolated from aquatic environment and diarrheal stool, showed that all of them produced a ß-lactamase with a pI of 5.4. Hybridization or amplification by PCR with a probe for blaTEM or primers for blaCARB gene families was negative. In this work, an environmental ampicillin-resistant strain from this sample, ME11762, isolated from a waterway in the west region of Argentina, was studied. The nucleotide sequence of the structural gene of the ß-lactamase was determined by bidirectional sequencing of a Sau3AI fragment belonging to this isolate. The gene encodes a new 288-amino-acid protein, designated CARB-7, that shares 88.5% homology with the CARB-6 enzyme; an overall 83.2% homology with PSE-4, PSE-1, CARB-3, and the Proteus mirabilis N29 enzymes; and 79% homology with CARB-4 enzyme. The gene for this ß-lactamase could not be transferred to Escherichia coli by conjugation. The nucleotide sequence of the flanking regions of the blaCARB-7 gene showed the occurrence of three 123-bp V. cholerae repeated sequences, all of which were found outside the predicted open reading frame. The upstream fragment of the blaCARB-7 gene shared 93% identity with a locus situated inside V. cholerae's chromosome 2. These results strongly suggest the chromosomal location of the blaCARB-7 gene, making this the first communication of a ß-lactamase gene located on the VCR island of the V. cholerae genome.

* Corresponding author. Mailing address: Servicio Antimicrobianos, INEI-ANLIS "Dr. Carlos G. Malbrán," Av. Velez Sarsfield 563 (1281), Buenos Aires, Argentina. Phone and fax: 54-11-4303-2812. E-mail: mgalas{at}anlis.gov.ar.

{dagger} Dedicated to the memory of our dear Ali.

{ddagger} Deceased.

Antimicrobial Agents and Chemotherapy, July 2002, p. 2162-2168, Vol. 46, No. 7
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.7.2162-2168.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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