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JAC Advance Access published online on July 8, 2008
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkn280
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© The Author 2008. 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

Original research

Analysis of tetracycline resistance tet(W) genes and their flanking sequences in intestinal Bifidobacterium species

Mohammed Salim Ammor{dagger}, Ana Belén Flórez, Pablo Álvarez-Martín, Abelardo Margolles and Baltasar Mayo*

Departamento de Microbiología y Bioquímica, Instituto de Productos Lácteos de Asturias (CSIC), Carretera de Infiesto s/n, 33300 Villaviciosa, Asturias, Spain

Received 18 April 2008; returned 12 May 2008; revised 29 May 2008; accepted 11 June 2008

* Corresponding author. Tel: +34-985892921; Fax: +34-985892933; E-mail: baltasar.mayo{at}ipla.csic.es

Objectives: The tet(W) gene provides tetracycline resistance to a wide range of anaerobic intestinal and ruminal bacteria, but little is known about the molecular organization of the tet(W) gene. The aim of this study was to gain new insights into the molecular organization of the tet(W) gene in bifidobacteria strains from humans.

Methods: A segment of DNA encompassing the whole tet(W) gene and its immediate upstream and downstream sequences was analysed in 10 representative strains of four Bifidobacterium species, of which two have been shown to be tetracycline-susceptible. The non-conserved flanking regions of the tet(W) gene were further analysed in six strains.

Results: All 10 strains share a core DNA domain of 2154 bp [starting 250 bp upstream of the tet(W) gene start codon and ending 13 bp before the stop codon] with 98% to 100% DNA identity. Except for Bifidobacterium animalis E43, all other strains further share 408 bp upstream and 70 bp downstream of the tet(W) gene. An insertion-like element of 736 bp was found to interrupt the tet(W) coding sequence in Bifidobacterium longum M21, which may be the reason for its tetracycline susceptibility. However, genetic events explaining the susceptible phenotype of B. longum LMG 13197T were not observed.

Conclusions: The tet(W) genes from all 10 strains shared 98% to 100% DNA and amino acid identity, though large variation was found in their flanking regions.

Key Words: bifidobacteria , antibiotic resistance , probiotics

{dagger} Present address. Centrale Laitière-DANONE, 3ème Étage, Tour A, Twin Center, Mâarif, 20000-Casablanca, Morocco


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