title: Two different tetracycline resistance mechanisms, plasmid-carried tet(L) and chromosomally located transposon-associated tet(M), coexist in Lactobacillus sakei Rits 9.
abstract: Lactobacillus sakei is extensively used as functional starter culture in fermented meat products. One of the safety criteria of a starter culture is the absence of potentially transferable antibiotic resistance determinants. However, tetracycline-resistant L. sakei strains have already been observed. In this paper, we show that tetracycline resistance in L. sakei Rits 9, a strain isolated from Italian Sola cheese made from raw milk, is mediated by a transposon-associated tet(M) gene coding for a ribosomal protection protein and a plasmid-carried tet(L) gene coding for a tetracycline efflux pump. pLS55, the 5-kb plasmid carrying the tet(L) gene, is highly similar to the pMA67 plasmid recently described for Paenibacillus larvae, a species pathogenic to honeybees. pLS55 could be transferred by electroporation into the laboratory strain L. sakei 23K. While the L. sakei 23K transformant containing pLS55 displayed an intermediate tetracycline resistance level (MIC, <32 microg/ml), L. sakei Rits 9, containing both tetracycline-resistant determinants, had a MIC of <256 microg/ml, suggesting that Tet L and Tet M confer different levels of resistance in L. sakei. Remarkably, in the absence of tetracycline, a basal expression of both genes was detected for L. sakei Rits 9. In addition, subinhibitory concentrations of tetracycline affected the expression patterns of tet(M) and tet(L) in different ways: the expression of tet(M) was induced only at high tetracycline concentrations, whereas the expression of tet(L) was up-regulated at lower concentrations. This is the first time that two different mechanisms conferring resistance to tetracycline are characterized for the same strain of a lactic acid bacterium.
authors.affiliation: Instituto de Productos Lácteos de Asturias (CSIC), Carretera de Infiesto s/n, 33300 Villaviciosa, Asturias, Spain.
authors.forename: Mohammed Salim
authors.initials: MS
authors.lastname: Ammor
authors.forename: Miguel
authors.initials: M
authors.lastname: Gueimonde
authors.forename: Morten
authors.initials: M
authors.lastname: Danielsen
authors.forename: Monique
authors.initials: M
authors.lastname: Zagorec
authors.forename: Angela H A M
authors.initials: AH
authors.lastname: van Hoek
authors.forename: Clara G
authors.initials: CG
authors.lastname: de Los Reyes-Gavilán
authors.forename: Baltasar
authors.initials: B
authors.lastname: Mayo
authors.forename: Abelardo
authors.initials: A
authors.lastname: Margolles
mesh_terms: D017920:Antiporters / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D015139:Blotting, Southern; D002611:Cheese / Q000382:microbiology; D017931:DNA Primers / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D007778:Lactobacillus / Q000235:genetics*; D046228:Microarray Analysis; D008969:Molecular Sequence Data; D010802:Phylogeny; D010957:Plasmids / Q000235:genetics; D017422:Sequence Analysis, DNA; D013753:Tetracycline Resistance / Q000235:genetics*
chemical_list: D017920:Antiporters; D001426:Bacterial Proteins; D017931:DNA Primers; D004251:DNA Transposable Elements; C083138:Tet M resistance protein, Bacteria; C079051:tetA protein, Bacteria