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Applied and Environmental Microbiology, October 2009, p. 6352-6360, Vol. 75, No. 19
0099-2240/09/$08.00+0     doi:10.1128/AEM.00470-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Intra- and Interspecies Conjugal Transfer of Tn916-Like Elements from Lactococcus lactis In Vitro and In Vivo{triangledown}

Joanna Boguslawska,1,{dagger} Joanna Zycka-Krzesinska,1 Andrea Wilcks,2 and Jacek Bardowski1*

Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, Poland,1 Department of Microbiology and Risk Assessment, National Food Institute, Technical University of Denmark, Mørkhøj Bygade 19, DK-2860 Søborg, Denmark2

Received 25 February 2009/ Accepted 30 July 2009

Tetracycline-resistant Lactococcus lactis strains originally isolated from Polish raw milk were analyzed for the ability to transfer their antibiotic resistance genes in vitro, using filter mating experiments, and in vivo, using germfree rats. Four of six analyzed L. lactis isolates were able to transfer tetracycline resistance determinants in vitro to L. lactis Bu2-60, at frequencies ranging from 10–5 to 10–7 transconjugants per recipient. Three of these four strains could also transfer resistance in vitro to Enterococcus faecalis JH2-2, whereas no transfer to Bacillus subtilis YBE01, Pseudomonas putida KT2442, Agrobacterium tumefaciens UBAPF2, or Escherichia coli JE2571 was observed. Rats were initially inoculated with the recipient E. faecalis strain JH2-2, and after a week, the L. lactis IBB477 and IBB487 donor strains were introduced. The first transconjugants were detected in fecal samples 3 days after introduction of the donors. A subtherapeutic concentration of tetracycline did not have any significant effect on the number of transconjugants, but transconjugants were observed earlier in animals dosed with this antibiotic. Molecular analysis of in vivo transconjugants containing the tet(M) gene showed that this gene was identical to tet(M) localized on the conjugative transposon Tn916. Primer-specific PCR confirmed that the Tn916 transposon was complete in all analyzed transconjugants and donors. This is the first study showing in vivo transfer of a Tn916-like antibiotic resistance transposon from L. lactis to E. faecalis. These data suggest that in certain cases food lactococci might be involved in the spread of antibiotic resistance genes to other lactic acid bacteria.

* Corresponding author. Mailing address: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, Poland. Phone: 48 22/659 70 72. Fax: 48 22/592 21 90. E-mail: jacek{at}ibb.waw.pl

{triangledown} Published ahead of print on 7 August 2009.

{dagger} Present address: Department of Biochemistry and Molecular Biology, The Medical Centre of Postgraduate Education, Marymoncka 99, 01-813 Warsaw, Poland.

Applied and Environmental Microbiology, October 2009, p. 6352-6360, Vol. 75, No. 19
0099-2240/09/$08.00+0     doi:10.1128/AEM.00470-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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