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Applied and Environmental Microbiology, July 2003, p. 3840-3848, Vol. 69, No. 7
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.7.3840-3848.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Bacterial Community Dynamics during Production of Registered Designation of Origin Salers Cheese as Evaluated by 16S rRNA Gene Single-Strand Conformation Polymorphism Analysis

Frédérique Duthoit,1 Jean-Jacques Godon,2 and Marie-Christine Montel1*

Laboratoire de Recherches Fromagères, INRA, 15000 Aurillac,1 Laboratoire de Biotechnologie de l'Environnement, INRA, 11100 Narbonne, France2

Received 13 November 2002/ Accepted 24 April 2003

Microbial dynamics during processing and ripening of traditional cheeses such as registered designation of origin Salers cheese, an artisanal cheese produced in France, play an important role in the elaboration of sensory qualities. The aim of the present study was to obtain a picture of the dynamics of the microbial ecosystem of RDO Salers cheese by using culture-independent methods. This included DNA extraction, PCR, and single-strand conformation polymorphism (SSCP) analysis. Bacterial and high-GC% gram-positive bacterial primers were used to amplify V2 or V3 regions of the 16S rRNA gene. SSCP patterns revealed changes during the manufacturing of the cheese. Patterns of the ecosystems of cheeses that were provided by three farmers were also quite different. Cloning and sequencing of the 16S rRNA gene revealed sequences related to lactic acid bacteria (Lactococcus lactis, Streptococcus thermophilus, Enterococcus faecium, Leuconostoc mesenteroides, Leuconostoc pseudomesenteroides, Lactobacillus plantarum, and Lactobacillus pentosus), which were predominant during manufacturing and ripening. Bacteria belonging to the high-GC% gram-positive group (essentially corynebacteria) were found by using specific primers. The present molecular approach can effectively describe the ecosystem of artisanal dairy products.

* Corresponding author. Mailing address: INRA-LRF, 36 rue de Salers, 15000 Aurillac, France. Phone: 33-4-71-45-64-10. Fax: 33-4-71-45-64-13. E-mail: cmontel{at}clermont.inra.fr.

Applied and Environmental Microbiology, July 2003, p. 3840-3848, Vol. 69, No. 7
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.7.3840-3848.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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