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Applied and Environmental Microbiology, August 2000, p. 3586-3591, Vol. 66, No. 8
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Real-Time Measurements of the Interaction between Single Cells of Listeria monocytogenes and Nisin on a Solid Surface

Birgitte Bjørn Budde* and Mogens Jakobsen

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

Received 14 January 2000/Accepted 10 May 2000

A method to obtain real-time measurements of the interactions between nisin and single cells of Listeria monocytogenes on a solid surface was developed. This method was based on fluorescence ratio-imaging microscopy and measurements of changes in the intracellular pH (pHi) of carboxyfluorescein succinimidyl ester-stained cells during exposure to nisin. Immobilized cells were placed in a chamber mounted on a microscope and attached to a high-precision peristaltic pump which allowed rapid changes in the nisin concentration. In the absence of nisin, the pHi of L. monocytogenes was almost constant (approximately pH 8.0) and independent of the external pH in the pH range from 5.0 to 9.0. In the presence of nisin, dissipation of the pH gradient (Delta pH) was observed, and this dissipation was both time and nisin concentration dependent. The dissipation of Delta pH resulted in cell death, as determined by the number of CFU. In the model system which we used the immobilized cells were significantly more resistant to nisin than the planktonic cells. The kinetics of Delta pH dissipation for single cells revealed a variable lag phase depending on the nisin concentration, which was followed by a very rapid decrease in pHi within 1 to 2 min. The differences in nisin sensitivity between single cells in a L. monocytogenes population were insignificant for cells grown to the stationary phase in a liquid laboratory substrate, but differences were observed for cells grown on an agar medium under similar conditions, which resulted in some cells having increased resistance to nisin.

* Corresponding author. Mailing address: Department of Dairy and Food Science, The Royal Veterinary and Agricultural University, Rolighedsvej 30, 4th floor, DK-1958 Frederiksberg C, Denmark. Phone: 45 35283284. Fax: 45 35283214. E-mail: bbb{at}kvl.dk.

Applied and Environmental Microbiology, August 2000, p. 3586-3591, Vol. 66, No. 8
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

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