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Applied and Environmental Microbiology, November 2001, p. 5113-5121, Vol. 67, No. 11
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.11.5113-5121.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Denaturing Gradient Gel Electrophoresis Analysis of the 16S rRNA Gene V1 Region To Monitor Dynamic Changes in the Bacterial Population during Fermentation of Italian Sausages

Luca Cocolin,1,* Marisa Manzano,1 Carlo Cantoni,2 and Giuseppe Comi1

Dipartimento di Scienze degli Alimenti, Facoltà di Agraria, Università degli studi di Udine, 33100 Udine,1 and Dipartimento di Scienze e Tecnologie Veterinarie per la Sicurezza degli Alimenti, Facoltà di Medicina Veterinaria, Università degli studi di Milano, 20121 Milan,2 Italy

Received 16 January 2001/Accepted 30 July 2001

In this study, a PCR-denaturing gradient gel electrophoresis (DGGE) protocol was used to monitor the dynamic changes in the microbial population during ripening of natural fermented sausages. The method was first optimized by using control strains from international collections, and a natural sausage fermentation was studied by PCR-DGGE and traditional methods. Total microbial DNA and RNA were extracted directly from the sausages and subjected to PCR and reverse transcription-PCR, and the amplicons obtained were analyzed by DGGE. Lactic acid bacteria (LAB) were present together with other organisms, mainly members of the family Micrococcaceae and meat contaminants, such as Brochothrix thermosphacta and Enterococcus sp., during the first 3 days of fermentation. After 3 days, LAB represented the main population, which was responsible for the acidification and proteolysis that determined the characteristic organoleptic profile of the Friuli Venezia Giulia fermented sausages. The PCR-DGGE protocol for studying sausage fermentation proved to be a good tool for monitoring the process in real time, and it makes technological adjustments possible when they are required.

* Corresponding author. Mailing address: Dipartimento di Scienze degli Alimenti, Via Marangoni 97, 33100 Udine, Italy. Phone: 0039/0432/590-746 or 0039/0432/590-730. Fax: 0039/0432/590-719. E-mail: luca.cocolin{at}dsa.uniud.it.

Applied and Environmental Microbiology, November 2001, p. 5113-5121, Vol. 67, No. 11
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.11.5113-5121.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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