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Applied and Environmental Microbiology, September 2004, p. 5644-5650, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5644-5650.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Raw Cow Milk Bacterial Population Shifts Attributable to Refrigeration

Véronique Lafarge,1,2* Jean-Claude Ogier,2 Victoria Girard,1 Véronique Maladen,1 Jean-Yves Leveau,3 Alexandra Gruss,2 and Agnès Delacroix-Buchet2

Unité Composition et Hygiène des Produits Laitiers, Agence Française de Sécurité Sanitaire des Aliments, Maisons Alfort,1 Unité de Recherches Laitières et Génétique Appliquée, Institut National de la Recherche Agronomique, Jouy en Josas,2 Unité Microbiologie Industrielle, Ecole Nationale Supérieure des Industries Agricoles et Alimentaires, Massy, France3

Received 11 February 2004/ Accepted 19 May 2004

We monitored the dynamic changes in the bacterial population in milk associated with refrigeration. Direct analyses of DNA by using temporal temperature gel electrophoresis (TTGE) and denaturing gradient gel electrophoresis (DGGE) allowed us to make accurate species assignments for bacteria with low-GC-content (low-GC%) (<55%) and medium- or high-GC% (>55%) genomes, respectively. We examined raw milk samples before and after 24-h conservation at 4°C. Bacterial identification was facilitated by comparison with an extensive bacterial reference database (~150 species) that we established with DNA fragments of pure bacterial strains. Cloning and sequencing of fragments missing from the database were used to achieve complete species identification. Considerable evolution of bacterial populations occurred during conservation at 4°C. TTGE and DGGE are shown to be a powerful tool for identifying the main bacterial species of the raw milk samples and for monitoring changes in bacterial populations during conservation at 4°C. The emergence of psychrotrophic bacteria such as Listeria spp. or Aeromonas hydrophila is demonstrated.

* Corresponding author. Mailing address: Unité Composition et Hygiène des Produits Laitiers, Agence Française de Sécurité Sanitaire des Aliments, 39-41 rue du 11 novembre 1918, 94700 Maisons Alfort, France. Phone: (33) 1-49-77-11-08. Fax: (33) 1-49-77-11-02. E-mail: v.lafarge{at}afssa.fr.

Applied and Environmental Microbiology, September 2004, p. 5644-5650, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5644-5650.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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