Parmigiano Reggiano cheese: evolution of cultivable and total lactic microflora and peptidase activities during manufacture and ripening

Parmigiano Reggiano is a Protected Designation of Origin, long-ripened cheese, made from cow's milk supplemented with natural whey starter, which thus contains a large microbial biodiversity. The aim of this study was to understand the population dynamics of the total lactic microflora throughout the manufacture and ripening of this cheese. Several approaches were combined to determine the quantitative changes in the different bacterial populations during 20 months of ripening of Parmigiano Reggiano cheeses from the same cheesemaking. Total and viable cells were enumerated after fluorescent labeling. Culturable bacteria were enumerated on different plate count agar media, including original media prepared from curd and ripened cheese. Six peptidase activities were quantified in curd and cheese samples free from cells. While the total bacterial cultivable population remained high and similar for the first six months, a decrease in viable starter lactic acid bacteria was observed during the first 48 h. The non-starter lactic acid bacteria populations, initially present in low numbers, began to grow after the brining and remained at high levels (about 10(7) CFU.g(-1)) for at least 10 months. During ripening, a strong decrease in the total bacterial population and a marked increase in 4 out of 6 peptidase activities were observed. In the external and internal zones of Parmigiano Reggiano cheese different trends in microbial growth, cell autolysis and peptidase activity were observed. This study gives for the first time a global view of the possible contribution of total, viable, cultivable and lysed bacterial cells throughout the ripening of Parmigiano Reggiano cheese.