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Journal of Clinical Microbiology, July 2003, p. 2940-2945, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.2940-2945.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Molecular Epidemiology and Characterization of Plasmid-Encoded ß-Lactamases Produced by Tunisian Clinical Isolates of Salmonella enterica Serotype Mbandaka Resistant to Broad-Spectrum Cephalosporins

Abdoulaye Makanera,1 Guillaume Arlet,2,3* Valérie Gautier,3 and Mohamed Manai1*

Laboratoire de Biochimie et Biologie Moléculaire, Faculté des Sciences de Tunis, Département de Biologie, Université de Tunis El Manar, 2092 Tunis, Tunisia,1 Service de Bactériologie-Hygiène, Hôpital Tenon, AP-HP,2 Laboratoire de Bactériologie, UPRES EA 2392, UFR Saint-Antoine,Paris, France3

Received 9 October 2002/ Returned for modification 21 January 2003/ Accepted 13 March 2003

We studied 31 clinical isolates of Salmonella enterica serotype Mbandaka resistant to broad-spectrum cephalosporins and recovered in Tunisia over a 5-year period. The transferability of this resistance was demonstrated by conjugation experiments. Thirty of the 31 isolates were positive in the double-disk synergy test. By isoelectric focusing analysis, all of the isolates were found to produce a band of ß-lactamase activity with a pI of 5.9. Three of these isolates produced an additional band with a pI of 7.6. PCR and DNA sequencing identified these ß-lactamases as TEM-4 and SHV-2a, respectively. The remaining isolate, highly resistant to ceftazidime but susceptible to cefepime, produced a ß-lactamase that focused at pI 7.8. No synergy was detected by the double-disk synergy test. Sequence analysis of the bla gene amplified by PCR showed that the plasmid-mediated AmpC-type enzyme was ACC-1a. Fingerprinting analysis by repetitive-element PCR and enterobacterial repeat intergenic consensus-PCR suggested that 29 of the 31 Salmonella serotype Mbandaka isolates belonged to the same clonal population.

* Corresponding author. Mailing address for Guillaume Arlet: Service de Bactériologie-Hygiène, Hôpital Tenon, 4 Rue de la Chine, 75970 Paris Cedex 20, France. Phone: 33 1 56 01 70 18. Fax: 33 1 56 01 61 08. E-mail: guillaume.arlet{at}tnn.ap-hop-paris.fr. Mailing address for Mohamed Manai: Laboratoire de Biochimie et Biologie Moléculaire, Faculté des Sciences de Tunis, Département de Biologie, Université de Tunis El Manar, 2092 El Manar, Tunis, Tunisia. Phone: 216 71 872 600. Fax: 216 71 885 480. E-mail: mohamedmanai{at}yahoo.fr.

Journal of Clinical Microbiology, July 2003, p. 2940-2945, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.2940-2945.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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