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Antimicrobial Agents and Chemotherapy, October 2008, p. 3589-3596, Vol. 52, No. 10
0066-4804/08/$08.00+0     doi:10.1128/AAC.00465-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Characterization of the New Metallo-β-Lactamase VIM-13 and Its Integron-Borne Gene from a Pseudomonas aeruginosa Clinical Isolate in Spain{triangledown}

Carlos Juan,1,2* Alejandro Beceiro,3 Olivia Gutiérrez,1,2 Sebastián Albertí,2,4 Margalida Garau,5 José L. Pérez,1,2 Germán Bou,3 and Antonio Oliver1,2,4

Servicio de Microbiología and Unidad de Investigación, Hospital Son Dureta, Palma de Mallorca, Spain,1 Instituto Universitario de Investigación en Ciencias de la Salud, Palma de Mallorca, Spain,2 Servicio de Microbiología, Complejo Hospitalario Juan Canalejo, La Coruña, Spain,3 Área de Microbiología, Universidad de las Islas Baleares, Palma de Mallorca, Spain,4 Área de Microbiología, Hospital Son Llàtzer, Palma de Mallorca, Spain5

Received 8 April 2008/ Returned for modification 25 June 2008/ Accepted 14 July 2008

During a survey conducted to evaluate the incidence of class B carbapenemase (metallo-β-lactamase [MBL])-producing Pseudomonas aeruginosa strains from hospitals in Majorca, Spain, five clinical isolates showed a positive Etest MBL screening test result. In one of them, strain PA-SL2, the presence of a new blaVIM derivative (blaVIM-13) was detected by PCR amplification with blaVIM-1-specific primers followed by sequencing. The blaVIM-13-producing isolate showed resistance to all β-lactams (except aztreonam), gentamicin, tobramycin, and ciprofloxacin. VIM-13 exhibited 93% and 88% amino acid sequence identities with VIM-1 and VIM-2, respectively. blaVIM-13 was cloned in parallel with blaVIM-1, and the resistance profile conferred was analyzed both in Escherichia coli and in P. aeruginosa backgrounds. Compared to VIM-1, VIM-13 conferred slightly higher levels of resistance to piperacillin and lower levels of resistance to ceftazidime and cefepime. VIM-13 and VIM-1 were purified in parallel as well, and their kinetic parameters were compared. The kcat/Km ratios for the antibiotics mentioned above were in good agreement with the MIC data. Furthermore, EDTA inhibited the activity of VIM-13 approximately 25 times less than it inhibited the activity of VIM-1. VIM-13 was harbored in a class 1 integron, along with a new variant (Ala108Thr) of the aminoglycoside-modifying enzyme encoding gene aacA4, which confers resistance to gentamicin and tobramycin. Finally, the VIM-13 integron was apparently located in the chromosome, since transformation and conjugation experiments consistently yielded negative results and the blaVIM-13 probe hybridized only with the genomic DNA.

* Corresponding author. Mailing address: Laboratorio de Microbiología, Unidad de Investigación, Hospital Son Dureta, C. Andrea Doria No. 55, Palma de Mallorca 07014, Spain. Phone: 34 971 175 334. Fax: 34 971 175 185. E-mail: cjuan{at}hsd.es

{triangledown} Published ahead of print on 21 July 2008.

Antimicrobial Agents and Chemotherapy, October 2008, p. 3589-3596, Vol. 52, No. 10
0066-4804/08/$08.00+0     doi:10.1128/AAC.00465-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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  • Strateva, T., Yordanov, D. (2009). Pseudomonas aeruginosa - a phenomenon of bacterial resistance. J Med Microbiol 58: 1133-1148 [Abstract] [Full Text]  


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