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Applied and Environmental Microbiology, May 2001, p. 2011-2020, Vol. 67, No. 5
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.5.2011-2020.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Characterization of Non-Starter Lactic Acid Bacteria from Italian Ewe Cheeses Based on Phenotypic, Genotypic, and Cell Wall Protein Analyses

M. De Angelis,1 A. Corsetti,1,* N. Tosti,2 J. Rossi,1 M. R. Corbo,3 and M. Gobbetti4

Dipartimento di Scienze degli Alimenti, Sezione di Microbiologia Agro-alimentare, Università degli Studi di Perugia,1 and Istituto di Ricerche sul Miglioramento Genetico delle Piante Foraggere, CNR Perugia,2 Perugia, Istituto di Produzioni e Preparazioni Alimentari, Facoltà di Agraria, Università degli Studi di Foggia, Foggia,3 and Dipartimento di Difesa delle Piante e Microbiologia Applicata, Università degli Studi di Bari, Bari,4 Italy

Received 9 August 2000/Accepted 16 January 2001

Non-starter lactic acid bacteria (NSLAB) were isolated from 12 Italian ewe cheeses representing six different types of cheese, which in several cases were produced by different manufacturers. A total of 400 presumptive Lactobacillus isolates were obtained, and 123 isolates and 10 type strains were subjected to phenotypic, genetic, and cell wall protein characterization analyses. Phenotypically, the cheese isolates included 32% Lactobacillus plantarum isolates, 15% L. brevis isolates, 12% L. paracasei subsp. paracasei isolates, 9% L. curvatus isolates, 6% L. fermentum isolates, 6% L. casei subsp. casei isolates, 5% L. pentosus isolates, 3% L. casei subsp. pseudoplantarum isolates, and 1% L. rhamnosus isolates. Eleven percent of the isolates were not phenotypically identified. Although a randomly amplified polymorphic DNA (RAPD) analysis based on three primers and clustering by the unweighted pair group method with arithmetic average (UPGMA) was useful for partially differentiating the 10 type strains, it did not provide a species-specific DNA band or a combination of bands which permitted complete separation of all the species considered. In contrast, sodium dodecyl sulfate-polyacrylamide gel electrophoresis cell wall protein profiles clustered by UPGMA were species specific and resolved the NSLAB. The only exceptions were isolates phenotypically identified as L. plantarum and L. pentosus or as L. casei subsp. casei and L. paracasei subsp. paracasei, which were grouped together. Based on protein profiles, Italian ewe cheeses frequently contained four different species and 3 to 16 strains. In general, the cheeses produced from raw ewe milk contained a larger number of more diverse strains than the cheeses produced from pasteurized milk. The same cheese produced in different factories contained different species, as well as strains that belonged to the same species but grouped in different RAPD clusters.

* Corresponding author. Mailing address: Dipartimento di Scienze degli Alimenti, Sezione di Microbiologia Agro-alimentare, Università degli Studi di Perugia, Via S. Costanzo, 06126 Perugia, Italy. Phone: 0039 (0)75-5857925. Fax: 0039 (0)75-32387. E-mail: corsetti{at}unipg.it.

Applied and Environmental Microbiology, May 2001, p. 2011-2020, Vol. 67, No. 5
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.5.2011-2020.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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