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JAC Advance Access originally published online on March 16, 2007
Journal of Antimicrobial Chemotherapy 2007 59(5):900-912; doi:10.1093/jac/dkm035
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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

Antimicrobial susceptibilities of Lactobacillus, Pediococcus and Lactococcus human isolates and cultures intended for probiotic or nutritional use

Ingo Klare1,*, Carola Konstabel1, Guido Werner1, Geert Huys2, Vanessa Vankerckhoven3,5, Gunnar Kahlmeter4, Bianca Hildebrandt1, Sibylle Müller-Bertling1, Wolfgang Witte1 and Herman Goossens3,5

1 Robert Koch Institute, Wernigerode Branch, Wernigerode, Germany 2 Laboratory of Microbiology, Faculty of Sciences, Ghent University, Ghent, Belgium 3 Laboratory of Medical Microbiology, University of Antwerp, Antwerp, Belgium 4 Department of Clinical Microbiology, Central Hospital, Växjö, Sweden 5 Department of Medical Microbiology, Leiden University Medical Center, Leiden, The Netherlands

Received 30 October 2006; returned 12 December 2006; revised 17 January 2007; accepted 22 January 2007


* Corresponding author. Tel: +49-3943-679-247; Fax: +49-3943-679-207; E-mail: i.klare{at}rki.de

Objectives: To determine MICs of 16 antimicrobials representing all major classes for 473 taxonomically well-characterized isolates of lactic acid bacteria (LAB) encompassing the genera Lactobacillus, Pediococcus and Lactococcus. To propose tentative epidemiological cut-off (ECOFF) values for recognizing intrinsic and acquired antimicrobial resistances in numerically dominant species.

Methods: On the basis of depositors' information, LAB were grouped in categories of probiotic, nutritional, probiotic or nutritional research, human and animal isolates and tested for their antibiotic susceptibilities by broth microdilution using LAB susceptibility test medium (LSM). Tentative ECOFFs were defined according to the recommendations of the European Committee on Antimicrobial Susceptibility Testing. Isolates showing acquired antimicrobial resistance(s) were selected for PCR-based detection of resistance gene(s) and in vitro conjugative transfer experiments.

Results: Tentative ECOFF values of 13 antibiotics were determined for up to 12 LAB species. Generally, LAB were susceptible to penicillin, ampicillin, ampicillin/sulbactam, quinupristin/dalfopristin, chloramphenicol and linezolid. LAB exhibited broad or partly species-dependent MIC profiles of trimethoprim, trimethoprim/sulfamethoxazole, vancomycin, teicoplanin and fusidic acid. Three probiotic Lactobacillus strains were highly resistant to streptomycin. Although erythromycin, clindamycin and oxytetracycline possessed high antimicrobial activities, 17 Lactobacillus isolates were resistant to one or more of these antibiotics. Eight of them, including six probiotic and nutritional cultures, possessed erm(B) and/or tet(W), tet(M) or unidentified members of the tet(M) group. In vitro intra- and interspecies filter-mating experiments failed to show transfer of resistance determinants.

Conclusions: Finding of acquired resistance genes in isolates intended for probiotic or nutritional use highlights the importance of antimicrobial susceptibility testing in documenting the safety of commercial LAB.

Keywords: lactic acid bacteria , antimicrobial susceptibility testing , broth microdilution , MIC ranges , epidemiological cut-off values


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