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Applied and Environmental Microbiology, February 2002, p. 756-764, Vol. 68, No. 2
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.2.756-764.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Frequency of Bacteriocin Resistance Development and Associated Fitness Costs in Listeria monocytogenes

A. Gravesen,* A.-M. Jydegaard Axelsen, J. Mendes da Silva,,{dagger} T. B. Hansen, and S. Knøchel

Department of Dairy and Food Science, Royal Veterinary and Agricultural University, Rolighedsvej 30, DK-1958 Frederiksberg, Denmark

Received 24 July 2001/ Accepted 15 November 2001

Bacteriocin-producing starter cultures have been suggested as natural food preservatives; however, development of resistance in the target organism is a major concern. We investigated the development of resistance in Listeria monocytogenes to the two major bacteriocins pediocin PA-1 and nisin A, with a focus on the variations between strains and the influence of environmental conditions. While considerable strain-specific variations in the frequency of resistance development and associated fitness costs were observed, the influence of environmental stress seemed to be bacteriocin specific. Pediocin resistance frequencies were determined for 20 strains and were in most cases ca. 10-6. However, two strains with intermediate pediocin sensitivity had 100-fold-higher pediocin resistance frequencies. Nisin resistance frequencies (14 strains) were in the range of 10-7 to 10-2. Strains with intermediate nisin sensitivity were among those with the highest frequencies. Environmental stress in the form of low temperature (10°C), reduced pH (5.5), or the presence of NaCl (6.5%) did not influence the frequency of pediocin resistance development; in contrast, the nisin resistance frequency was considerably reduced (<5 x 10-8). Pediocin resistance in all spontaneous mutants was very stable, but the stability of nisin resistance varied. Pediocin-resistant mutants had fitness costs in the form of reduction down to 44% of the maximum specific growth rate of the wild-type strain. Nisin-resistant mutants had fewer and less-pronounced growth rate reductions. The fitness costs were not increased upon applying environmental stress (5°C, 6.5% NaCl, or pH 5.5), indicating that the bacteriocin-resistant mutants were not more stress sensitive than the wild-type strains. In a saveloy-type meat model at 5°C, however, the growth differences seemed to be negligible. The applicational perspectives of the results are discussed.

* Corresponding author. Mailing address: Department of Dairy and Food Science, Royal Veterinary and Agricultural University, Rolighedsvej 30, DK-1958 Frederiksberg, Denmark. Phone: 45-3528-3272. Fax: 45-3528-3231. E-mail: alg{at}kvl.dk.

{dagger} Present address: Novo Nordisk Engineering, Krogshøjvej 55, 2880 Bagsværd, Denmark.

Applied and Environmental Microbiology, February 2002, p. 756-764, Vol. 68, No. 2
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.2.756-764.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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