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Applied and Environmental Microbiology, April 2004, p. 1883-1894, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.1883-1894.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Study of the Ecology of Fresh Sausages and Characterization of Populations of Lactic Acid Bacteria by Molecular Methods

Luca Cocolin,1* Kalliopi Rantsiou,1 Lucilla Iacumin,1 Rosalinda Urso,1 Carlo Cantoni,2 and Giuseppe Comi1

Dipartimento di Scienze degli Alimenti, Università degli studi di Udine, 33100 Udine,1 Dipartimento di Scienze e Tecnologie Veterinarie per la Sicurezza degli Alimenti, Università degli studi di Milano, 20121 Milan, Italy2

Received 2 July 2003/ Accepted 24 November 2003

In this study, a polyphasic approach was used to study the ecology of fresh sausages and to characterize populations of lactic acid bacteria (LAB). The microbial profile of fresh sausages was monitored from the production day to the 10th day of storage at 4°C. Samples were collected on days 0, 3, 6, and 10, and culture-dependent and -independent methods of detection and identification were applied. Traditional plating and isolation of LAB strains, which were subsequently identified by molecular methods, and the application of PCR-denaturing gradient gel electrophoresis (DGGE) to DNA and RNA extracted directly from the fresh sausage samples allowed the study in detail of the changes in the bacterial and yeast populations during storage. Brochothrix thermosphacta and Lactobacillus sakei were the main populations present. In particular, B. thermosphacta was present throughout the process, as determined by both DNA and RNA analysis. Other bacterial species, mainly Staphylococcus xylosus, Leuconostoc mesenteroides, and L. curvatus, were detected by DGGE. Moreover, an uncultured bacterium and an uncultured Staphylococcus sp. were present, too. LAB strains isolated at day 0 were identified as Lactococcus lactis subsp. lactis, L. casei, and Enterococcus casseliflavus, and on day 3 a strain of Leuconostoc mesenteroides was identified. The remaining strains isolated belonged to L. sakei. Concerning the yeast ecology, only Debaryomyces hansenii was established in the fresh sausages. Capronia mansonii was initially present, but it was not detected after the first 3 days. At last, L. sakei isolates were characterized by randomly amplified polymorphic DNA PCR and repetitive DNA element PCR. The results obtained underlined how different populations took over at different steps of the process. This is believed to be the result of the selection of the particular population, possibly due to the low storage temperature employed.

* Corresponding author. Mailing address: Dipartimento di Scienze degli Alimenti, via Marangoni 97, 33100 Udine, Italy. Phone: 39 0432 590 759. Fax: 39 0432 590 719. E-mail: lscocolin{at}uniud.it.

Applied and Environmental Microbiology, April 2004, p. 1883-1894, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.1883-1894.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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