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Applied and Environmental Microbiology, July 2008, p. 4491-4497, Vol. 74, No. 14
0099-2240/08/$08.00+0     doi:10.1128/AEM.00255-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Listeria monocytogenes EGD-e Biofilms: No Mushrooms but a Network of Knitted Chains{triangledown}

Aurélie Rieu,4 Romain Briandet,3 Olivier Habimana,3 Dominique Garmyn,1,2 Jean Guzzo,4 and Pascal Piveteau1,2*

Université de Bourgogne, UMR 1229, F-21000 Dijon, France,1 INRA, UMR 1229, F-21000 Dijon, France,2 UMR 763, Unité Mixte de Recherche en Bioadhesion et Hygiène des Matériaux, INRA-AgroParisTech, F-91300 Massy, France,3 Laboratoire de Recherche En Vigne et Vin, Université de Bourgogne, IUVV, F-21000 Dijon, France4

Received 29 January 2008/ Accepted 13 May 2008

Listeria monocytogenes is a food pathogen that can attach on most of the surfaces encountered in the food industry. Biofilms are three-dimensional microbial structures that facilitate the persistence of pathogens on surfaces, their resistance toward antimicrobials, and the final contamination of processed goods. So far, little is known about the structural dynamics of L. monocytogenes biofilm formation and its regulation. The aims of this study were, by combining genetics and time-lapse laser-scanning confocal microscopy (LSCM), (i) to characterize the structural dynamics of L. monocytogenes EGD-e sessile growth in two nutritional environments (with or without a nutrient flow), and (ii) to evaluate the possible role of the L. monocytogenes agr system during biofilm formation by tracking the spatiotemporal fluorescence expression of a green fluorescent protein (GFP) reporter system. In the absence of nutrient flow (static conditions), unstructured biofilms composed of a few layers of cells that covered the substratum were observed. In contrast, when grown under dynamic conditions, L. monocytogenes EGD-e biofilms were highly organized. Indeed, ball-shaped microcolonies were surrounded by a network of knitted chains. The spatiotemporal tracking of fluorescence emitted by the GFP reporter system revealed that agr expression was barely detectable under static conditions, but it progressively increased during 40 h under dynamic conditions. Moreover, spatial analysis revealed that agr was expressed preferentially in cells located outside the microcolonies. Finally, the in-frame deletion of agrA, which encodes a transcriptional regulator, resulted in a decrease in initial adherence without affecting the subsequent biofilm development.

* Corresponding author. Mailing address: Laboratoire de Microbiologie du Sol et de l'Environnement UMR UB/INRA 1229, 17 Rue de Sully, BP86510, F-21065 Dijon, France. Phone: 33 3 80 69 34 32. Fax: 33 3 80 69 32 24. E-mail: piveteau{at}u-bourgogne.fr

{triangledown} Published ahead of print on 23 May 2008.

Applied and Environmental Microbiology, July 2008, p. 4491-4497, Vol. 74, No. 14
0099-2240/08/$08.00+0     doi:10.1128/AEM.00255-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Vejborg, R. M., Klemm, P. (2009). Cellular chain formation in Escherichia coli biofilms. Microbiology 155: 1407-1417 [Abstract] [Full Text]  


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