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JAC Advance Access published online on August 17, 2007
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkm293
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© The Author 2007. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Nucleotide sequence and transfer properties of two novel types of Actinobacillus pleuropneumoniae plasmids carrying the tetracycline resistance gene tet(H)

Mónica Blanco1, Kristina Kadlec2, César B. Gutiérrez Martín3, Ana Judith Martín de la Fuente3, Stefan Schwarz2 and Jesús Navas1,*

1 Departamento de Biología Molecular (Unidad Asociada al Centro de Investigaciones Biológicas, C.S.I.C.), Facultad de Medicina, Universidad de Cantabria, 39011 Santander, Spain 2 Institut für Tierzucht, Bundesforschungsanstalt für Landwirtschaft (FAL), Höltystr. 10, 31535 Neustadt-Mariensee, Germany 3 Departamento de Sanidad Animal, Unidad de Microbiología e Inmunología, Facultad de Veterinaria, Universidad de León, 24007 León, Spain

Received 13 June 2007; returned 2 July 2007; revised 9 July 2007; accepted 11 July 2007

* Corresponding author. Tel: +34-942-201943; Fax: +34-942-201945; E-mail: navasj{at}unican.es

Objectives: To analyse the sequence and transfer properties of two tetracycline resistance plasmids found in clinical isolates of Actinobacillus pleuropneumoniae in order to assess their role in the spread of tetracycline resistance.

Methods: The plasmids designated p9956 and p12494 were purified from A. pleuropneumoniae and completely sequenced. The transfer properties of both plasmids were evaluated by electroporation and/or conjugation into Pasteurella multocida and Escherichia coli.

Results: Both plasmids showed a modular structure consisting of three regions involved in mobilization, tetracycline resistance or replication. The mobilization regions included the mobA gene, encoding a relaxase, a protein involved in plasmid transfer. The tetracycline resistance regions were closely related and consisted of the tet(H) gene and its repressor gene tetR. The tetracycline resistance phenotype was transferred successfully to P. multocida and in the case of p9956 also to E. coli by electroporation of the plasmids. Moreover, plasmid p9956 could be mobilized in E. coli with the assistance of RP4 conjugal transfer functions.

Conclusions: For the first time, the complete sequences of two tet(H)-carrying plasmids from A. pleuropneumoniae were determined. These two plasmids differed from one another and from known tet(H)-carrying plasmids from Pasteurella or Mannheimia spp. Structural analysis confirmed that these plasmids consisted of segments that have been previously detected in members of the families Pasteurellaceae and Enterobacteriaceae.

Key Words: respiratory tract infections , antimicrobial resistance , gene transfer , mobilization , interspecies transfer


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