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Applied and Environmental Microbiology, April 2007, p. 2118-2127, Vol. 73, No. 7
0099-2240/07/$08.00+0     doi:10.1128/AEM.01744-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Historical and Contemporary NaCl Concentrations Affect the Duration and Distribution of Lag Times from Individual Spores of Nonproteolytic Clostridium botulinum{triangledown}

Martin D. Webb, Carmen Pin, Michael W. Peck, and Sandra C. Stringer*

Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, United Kingdom

Received 25 July 2006/ Accepted 21 January 2007

In this study we determined the effect of NaCl concentration during sporulation (0 or 3.0% [wt/vol] added NaCl) and subsequent growth (0 or 2.0% [wt/vol] added NaCl) on the distributions of times associated with various stages of the lag phase of individual spores of nonproteolytic Clostridium botulinum strain Eklund 17B. The effects of NaCl on the probability of germination and the probability of subsequent growth were also determined. Spore populations exhibited considerable heterogeneity at all stages of lag phase for each condition tested. Germination time did not correlate strongly with the times for later stages in the lag phase, such as outgrowth and doubling time. Addition of NaCl to either the sporulation or growth media increased the mean times for, and variability of, all the measured stages of the lag phase (germination, emergence, time to one mature cell, and time to first doubling). There was a synergistic interaction between the inhibitory effects of NaCl in the sporulation medium and the inhibitory effects of NaCl in the subsequent growth medium on the total lag time and each of its stages. Addition of NaCl to either the sporulation medium or the growth medium reduced both the probability of germination and the probability of a germinated spore developing into a mature cell, but the interaction was not synergistic. Spores formed in medium with added NaCl were not better adapted to subsequent growth in suboptimal osmotic conditions than spores formed in medium with no added NaCl were. Knowledge of the distribution of lag times for individual spores and quantification of the biovariability within lag time distributions may provide insight into the underlying mechanisms and can be used to improve predictions of growth in food and to refine risk assessments.

* Corresponding author. Mailing address: Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, United Kingdom. Phone: (44)1603 255000. Fax: (44)1603 255288. E-mail: sandra.stringer{at}bbsrc.ac.uk.

{triangledown} Published ahead of print on 2 February 2007.

Applied and Environmental Microbiology, April 2007, p. 2118-2127, Vol. 73, No. 7
0099-2240/07/$08.00+0     doi:10.1128/AEM.01744-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Stringer, S. C., Webb, M. D., Peck, M. W. (2009). Contrasting Effects of Heat Treatment and Incubation Temperature on Germination and Outgrowth of Individual Spores of Nonproteolytic Clostridium botulinum Bacteria. Appl. Environ. Microbiol. 75: 2712-2719 [Abstract] [Full Text]  
  • Pin, C., Baranyi, J. (2008). Single-Cell and Population Lag Times as a Function of Cell Age. Appl. Environ. Microbiol. 74: 2534-2536 [Abstract] [Full Text]  


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