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Antimicrobial Agents and Chemotherapy, June 2004, p. 2344-2345, Vol. 48, No. 6
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.6.2344-2345.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

LETTER TO THE EDITOR

Emergence of the Extended-Spectrum ß-Lactamase GES-1 in a Pseudomonas aeruginosa Strain from Brazil: Report from the SENTRY Antimicrobial Surveillance Program


   LETTER
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 Letter
References
 
The extended-spectrum ß-lactamase (ESBL) GES-1 was initially described from a Klebsiella pneumoniae strain in 1998 (5). Since then, two closely related enzymes, IBC-1 and GES-2, have also been described (4, 6). A GES-1-producing K. pneumoniae strain has been reported from a French hospital, although the patient had just been transferred from French Guiana, South America. blaGES-1 has also been found in a Pseudomonas aeruginosa strain isolated in another French medical center (3) and in K. pneumoniae in Lisbon, Portugal (2).

The GES-1-producing P. aeruginosa, isolate 48-8896, was recovered from a 63-year-old female who had a hysterectomy (São Paulo, Brazil) and developed a wound infection while receiving ceftriaxone, amikacin, and metronidazole. Blood cultures were drawn and were positive for P. aeruginosa. Polymyxin B plus vancomycin was started empirically and the infection was eradicated, but the patient died after being hospitalized in the intensive care unit for 3 months. Isolate 48-8896 showed resistance to imipenem MIC, >8 µg/ml), meropenem (MIC, >8 µg/ml), ceftazidime (MIC, >16 µg/ml), cefepime (MIC, >16 µg/ml), piperacillin-tazobactam (MIC, >64 µg/ml), and all antimicrobial agents evaluated (including aminoglycosides and quinolones), except for polymyxin B (MIC, <=1 µg/ml). Phenotypic detection of ESBL was determined to be positive using the (ceftazidime/ceftazidime-clavulanic acid and cefepime/cefepime-clavulanic acid) Etest strips (AB BIODISK, Solna, Sweden).

Since several ß-lactamase genes are part of gene cassettes that are class 1 integron mediated and most of them contain an aminoglycoside acetyltransferase gene cassette, primers located in the 5' conserved segment region (5'-CS) (5'-CCAAGCTCTCGGGTAACATC-3') and in the flanking region of the aacA4 gene cassette (5'-AACTTGCGAGCGATCCGATG-3') were used. PCR amplification and subsequent sequencing analyses of the 1,211-bp PCR product showed a blaGES-1 in the first position of a class 1 integron (Fig. 1).



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  		FIG. 1. Schematic representations of different blaGES-1-containing integrons (a, b, c). The location of the three G residues found in the P2 of P. aeruginosa 48-8896 is highlighted. The 59 bp are represented with black dots.

 
The integron harboring blaGES-1 showed a similar adjacent and upstream context as described in the first report of GES-1 (K. pneumoniae ORI-1) (5). Beyond blaGES-1 was intI1, encoding the integrase of a class 1 integron. Between blaGES-1 and intI1 was an attI1 site and two promoters (P2 and Pant). However, the P2 promoter appears to have an absence of three G residues, and the space between the –35 and –10 sequences is 14 bp (Fig. 1). The secondary promoter (P2) is created by the insertion of the three Gs, increasing the space between the –35 and –10 regions to 17 bp (1). This evidence suggests that P2 was probably not contributing to the transcription of blaGES-1, as previously described in other integrons harboring this ESBL gene (3, 5). As determined in this study, beyond the blaGES-1 integron lies a chloramphenicol-acetyltransferase gene cassette, catB8, followed by the 3'-CS region of the class 1 integron, which is unique among blaGES-1 genes.

The detection of a GES-1-producing P. aeruginosa isolate in Latin America (Brazil) was a very worrisome finding, since it heralds the possibility for the emergence and future dissemination of new GES derivatives with broader spectrums of hydrolyses, such as the carbapenem-hydrolyzing enzyme GES-2 (6).


   ACKNOWLEDGMENTS
 
The SENTRY program was funded by an educational research grant from Bristol-Myers Squibb.


   REFERENCES
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 Letter
 References
 

  1. Collis, C. M., and R. M. Hall. 1995. Expression of antibiotic resistance genes in the integrated cassettes of integrons. Antimicrob. Agents Chemother. 39:155-162.[Abstract]
  2. Duarte, A., F. Boavida, F. Grosso, M. Correia, L. M. Lito, and J. M. Cristino. 2003. Outbreak of GES-1-lactamase-producing multidrug-resistant Klebsiella pneumoniae in a university hospital in Lisbon, Portugal. Antimicrob. Agents Chemother. 47:1481-1482.[Free Full Text]
  3. Dubois, V., L. Poirel, C. Marie, C. Arpin, P. Nordmann, and C. Quentin. 2002. Molecular characterization of a novel class 1 integron containing blaGES-1 and a fused product of aac3-Ib/aac6'-Ib' gene cassettes in Pseudomonas aeruginosa. Antimicrob. Agents Chemother. 46:638-645.[Abstract/Free Full Text]
  4. Giakkoupi, P., L. Tzouveleskis, A. Tsakris, V. Loukova, D. Sofianou, and E. Tzelepi. 2004. IBC-1, a novel integron-associated class A ß-lactamase with extended-spectrum properties produced by an Enterobacter cloacae clinical strain. Antimicrob. Agents Chemother. 44:2247-2253.
  5. Poirel, L., I. Le Thomas, T. Naas, A. Karim, and P. Nordmann. 2000. Biochemical sequence analyses of GES-1, a novel class A extended-spectrum ß-lactamase, and the class 1 integron In52 from Klebsiella pneumoniae. Antimicrob. Agents Chemother. 44:622-632.[Abstract/Free Full Text]
  6. Poirel, L., G. F. Weldhagen, T. Naas, C. De Champs, M. G. Dove, and P. Nordmann. 2001. GES-2, a class A ß-lactamase from Pseudomonas aeruginosa with increased hydrolysis of imipenem. Antimicrob. Agents Chemother. 45:2598-2603.[Abstract/Free Full Text]
Mariana Castanheira
Rodrigo E. Mendes
Timothy R. Walsh*
Department of Pathology & Microbiology
University of Bristol
Bristol, United Kingdom

Ana C. Gales
Disciplina de Doencas Infecciosas e Parasitarias
Universidade Federal de Sao Paulo
Sao Paulo, Brazil

Ronald N. Jones
The JONES Group/JMI Laboratories
North Liberty, Iowa

* Phone: 44 117 928-7522, Fax: 44 117 928-7896, E-mail: t.r.walsh{at}bristol.ac.uk

Antimicrobial Agents and Chemotherapy, June 2004, p. 2344-2345, Vol. 48, No. 6
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.6.2344-2345.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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