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JAC Advance Access published online on April 25, 2003
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkg257
© 2003 by The British Society for Antimicrobial Chemotherapy
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© 2003 The British Society for Antimicrobial Chemotherapy

Brief report

Molecular analysis of the plasmid-borne aacA/aphD resistance gene region of coagulase-negative staphylococci from chickens

Carla C. Lange 1, Christiane Werckenthin 2, Stefan Schwarz 3*

1 Institut für Tierzucht der Bundesforschungsanstalt für Landwirtschaft (FAL), Höltystr. 10, 31535 Neustadt-Mariensee, Germany; Empresa Brasileira de Pesquisa Agropecuária, Rua Eugênio do Nascimento 610, 36038-330 Juiz de Fora-MG, Brazil
2 Institut für Tierzucht der Bundesforschungsanstalt für Landwirtschaft (FAL), Höltystr. 10, 31535 Neustadt-Mariensee, Germany; Institut für Medizinische Mikrobiologie, Seuchen- und Infektionsmedizin, Ludwig-Maximilians-Universität München, Veterinärstr. 13, 80539 München, Germany
3 Institut für Tierzucht der Bundesforschungsanstalt für Landwirtschaft (FAL), Höltystr. 10, 31535 Neustadt-Mariensee, Germany

* Corresponding author. E-mail: stefan.schwarz{at}fal.de.

Received 3 February 2003 ; revised 20 February 2003 ; accepted 13 March 2003

Abstract

Objectives: The aim of this study was to analyse selected coagulase-negative staphylococci from chickens for the genetic basis of plasmid-borne resistance to gentamicin, tobramycin and kanamycin, and also for the presence of Tn4001-like elements.

Methods: Three staphylococcal strains, two Staphylococcus warneri, and one Staphylococcus sciuri, were included in this study. The gene aacA/aphD coding for a bifunctional enzyme that mediates resistance to gentamicin, tobramycin and kanamycin was detected by hybridization with a specific probe. Plasmid location of this gene was also confirmed by hybridization and conjugation. The resistance gene and its adjacent regions were cloned and sequenced.

Results: Three different types of Tn4001-like elements in which the IS256 elements were largely truncated and replaced by IS257 elements were identified on large conjugative and non-conjugative plasmids of 33-43 kb in the staphylococcal strains from chickens. Seven different types of IS257-analogous insertion sequences were identified.

Conclusions: Resistance to gentamicin, tobramycin and kanamycin in three staphylococcal strains from chickens was mediated by plasmid-borne aacA/aphD genes located on structurally modified Tn4001-like elements. In one of the three plasmids studied, the arrangement of the elements in the aacA/aphD resistance gene area closely resembled that on plasmids pSH6, pSK41 and pUW3626 from Staphylococcus aureus of human origin.

Keywords: aminoglycoside resistance, aacA-aphD gene, coagulase-negative staphylococci, chicken, plasmid
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