Canestrato Pugliese cheeses were produced from raw ewes’ milk R and R cheeses , pasteurized ewes’ milk P cheese . Ž . Ž . II and by heating the curd in hot whey according to a traditional protocol T cheese . R differed from R cheese mainly by having been produced from raw milk with a higher number of somatic cells, 950.000 vs. 750.000 mly1 , respectively. II Compared to P and T cheeses, R and R cheeses had a higher concentration one or two orders of magnitude of Ž . cheese-related bacteria such as adventitious mesophilic lactobacilli, enterococci and staphylococci. At the end of ripening, all cheeses contained less than 1.0 log cfu gy1 of total and fecal coliforms, and Escherichia coli and Staphylococcus aureus were not detected. As shown by phenotypic identification and RAPD-PCR, R cheese contained the largest number of mesophilic lactobacilli species and the greatest diversity of strains within the Lactobacillus plantarum species. Primary proteolysis did not differ appreciably among the cheeses. On the contrary, both urea-PAGE and the RP-HPLC analyses of the water-soluble N fractions showed the more complex profiles in cheeses produced by raw milks. R and R II cheeses had the highest values of water-soluble Nrtotal N ca. 30% and the highest concentration of total free amino acids ca. 40 mg Ž . Ž y1 g which approached or exceeded those reported for Italian cheeses with very high level of proteolysis during ripening. . The main differences between R–R II and P–T cheeses were the concentrations of aspartic acid, proline, alanine, isoleucine, histidine and lysine. The water-soluble extracts of R and R II cheeses contained levels of amino-, imino- and di-peptidase activities, which were about twice those found in P and T cheeses. Cheeses differed slightly in the concentration of total free fatty acids that ranged between 1673 and 1651 mg kgy1 in R and R II cheeses, and 1397 and 1334 mg kgy1 in P and T cheeses. Butyric, caproic, capric, palmitic, oleic and linoleic acids were found at the highest concentrations.