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Applied and Environmental Microbiology, May 2007, p. 2839-2846, Vol. 73, No. 9
0099-2240/07/$08.00+0     doi:10.1128/AEM.02267-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

First Evidence of Division and Accumulation of Viable but Nonculturable Pseudomonas fluorescens Cells on Surfaces Subjected to Conditions Encountered at Meat Processing Premises{triangledown}

Sophie Peneau, Danielle Chassaing, and Brigitte Carpentier*

Agence Française de Sécurité Sanitaire des Aliments, Laboratoire d'Etudes et de Recherches sur la Qualité des Aliments et sur les Procédés Agro-Alimentaires, Maisons-Alfort, France

Received 26 September 2006/ Accepted 21 February 2007

Cleaning and disinfection of open surfaces in food industry premises leave some microorganisms behind; these microorganisms build up a resident flora on the surfaces. Our goal was to explore the phenomena involved in the establishment of this biofilm. Ceramic coupons were contaminated, once only, with Pseudomonas fluorescens suspended in meat exudate incubated at 10°C. The mean adhering population after 1 day was 102 CFU·cm–2 and 103 total cells·cm–2, i.e., the total number of cells stained by DAPI (4',6'-diamidino-2-phenylindole). The coupons were subjected daily to a cleaning product, a disinfectant, and a further soiling with exudate. The result was a striking difference between the numbers of CFU, which reached 104 CFU·cm–2, and the numbers of total cells, which reached 2 x 106 cells·cm–2 in 10 days. By using hypotheses all leading to an overestimation of the number of dead cells, we showed that the quantity of nonculturable cells (DAPI-positive cells minus CFU) observed cannot be accounted for as an accumulation of dead cells. Some nonculturable cells are therefore dividing on the surface, although cell division is unable to continue to the stage of macrocolony formation on agar. The same phenomenon was observed when only a chlorinated alkaline product was used and the number of cells capable of reducing 5-cyano-2,3-ditolyl tetrazolium chloride was close to the number of total cells, confirming that most nonculturable cells are viable but nonculturable. Furthermore, the daily shock applied to the cells does not prompt them to enter a new lag phase. Since a single application of microorganisms is sufficient to produce this accumulation of cells, it appears that the phenomenon is inevitable on open surfaces in food industry premises.

* Corresponding author. Mailing address: Agence Française de Sécurité Sanitaire des Aliments, Laboratoire d'Etudes et de Recherches sur la Qualité des Aliments et sur les Procédés Agro-Alimentaires, Maisons-Alfort, France. Phone: 33(0)1 49 77 26 46. Fax: 33(0)1 43 68 97 62. E-mail: b.carpentier{at}afssa.fr

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

Applied and Environmental Microbiology, May 2007, p. 2839-2846, Vol. 73, No. 9
0099-2240/07/$08.00+0     doi:10.1128/AEM.02267-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Quiros, C., Herrero, M., Garcia, L. A., Diaz, M. (2009). Quantitative Approach to Determining the Contribution of Viable-but-Nonculturable Subpopulations to Malolactic Fermentation Processes. Appl. Environ. Microbiol. 75: 2977-2981 [Abstract] [Full Text]  


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