This paper describes the results of the characterization of a traditional Slovak cheese called “May bryndza” with regard to the profiles of volatile organic compounds and lactic acid bacteria. Samples of “May bryndza“ cheese produced solely from unpasteurized ewe's milk were collected from 4 different Slovak farms, and samples of the cheese produced from a mixture of 2 types of milk (raw ewe's and pasteurized cow's milk) were collected from 3 different Slovak industrial dairies. There were 15 compounds detected and identified by the electronic nose. The impact of the kind of milk and the kind of dairy on the aroma profile of the product was not confirmed by PCA. The compounds with the highest relative contents in samples were acetoin (2.59%–24.55%), acetic acid (6.69%–13.39%), methoxy-phenyl-oxime (4.49%–8.52%), butanoic acid (1.89%–5.67%), and 2,3-butanediol (0.98%–4.08%), which were determined with gas chromatography. A total of 1533 isolates of LAB were obtained from the “May bryndza” cheese samples. Four families, five genera, and 19 species were identified with mass spectrometry, and isolated bacteria, both from the farm and industry dairies were the most frequently found to belong to Lactococcus lactis subsp. lactis.
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