Application of Enterococcus faecium KE82, an Enterocin A-B-P-Producing Strain, as an Adjunct Culture Enhances Inactivation of Listeria monocytogenes during Traditional Protected Designation of Origin Galotyri Processing

Abstract

Abstract: The ability of the enterocin A-B-P-producing Enterococcus faecium KE82 adjunct strain to inactivate Listeria monocytogenes during protected designation of origin Galotyri processing was evaluated. Three trials were conducted with artisan cheeses made from traditionally "boiled" (85°C) ewe's milk. The milk was cooled at 42°C and divided in two treatments. A1 milk was inoculated with Streptococcus thermophilus ST1 and Lactococcus lactis subsp. cremoris M78, and A2 was inoculated with the basic starter ST1+M78 plus KE82 (step 1). All milks were fermented at 20 to 22°C for 24 h (step 2), and the curds were drained at 12°C for 72 h (step 3) and then salted with 1.5 to 1.8% salt to obtain the fresh Galotyri cheeses (step 4). These fresh cheeses were then ripened at 4°C for 30 days (step 5). Because artificial listerial contamination in the dairy plant was prohibited, samples of A1 and A2 cheese milk (200 mL) or curd (200 g) were collected after steps 1 through 5, inoculated with L. monocytogenes 10 (3 to 4 log CFU/mL or g), incubated at 37, 22, 12, and 4°C for predefined periods, and analyzed for microbial levels and pH. L. monocytogenes levels declined in all cheese curd portions contaminated after steps 2 through 5 (pH 4.36 to 4.84) when stored at 4 or 12°C for 15 days. The final net reductions in Listeria populations were 2.00-, 1.07-, 0.54-, and 0.61-log greater in the A2 than in the A1 curd portions after steps 2, 3, 4, and 5, respectively. In step 1, conducted to simulate the whole cheese milk fermentation process, L. monocytogenes levels declined by 1.47 log CFU/mL more in the A2 than in the A1 milk portions after 72 h at 22°C; however, slight growth (0.6 log CFU/mL) occurred during the first 6 h at 37°C. E. faecium KE82 was compatible with the starter culture and enhanced inactivation of L. monocytogenes during all steps of Galotyri cheese processing. The antilisterial effects of the combined acid and enterocin were the weakest in the fermenting milks, the strongest in the unsalted fermented curds, and declined again in the salted fresh cheeses.

Keywords: Enterococcus faecium; Listeria monocytogenes; Enterocin A-B-P; Galotyri cheese.