Determination of antilisterial effect of some microbial isolates from traditional zlatar cheese during the fermentation of soft white cheese

Abstract

The aim of this study was to select autochthonous lactic acid bacteria (LAB) isolates with antilisterial activity from Zlatar cheese and to evaluate the ability of selected LAB to control Listeria monocytogenes growth during soft white cheese production. The genotype characterization of isolated LAB (n = 93) was done using PCR method by 16S rRNA sequencing. In this way, the following isolates were detected: Lactococcus lactis ssp. lactis (40 isolates), Enterococcus faecalis (30), Lactobacillus plantarum (12), Leuconostoc mesenteroides ssp. mesenteroides (3) Lactobacillus garvieae (3), Lactobacillus curvatus (2), Lactobacillus casei (1), Enterococcus faecium (1) and Staphylococcus hominis (1). Each isolated LAB was tested for bacteriocin-producing ability. It was determined that two LAB isolates had bactericidal properties: Lactococcus lactis ssp. lactis SRB/ZS/094 and Enterococcus faecalis SRB/ZS/090. Semi-purified of enterococcal bacteriocin (enterocin) was isolated using precipitation procedures with ammonium sulphate. Its properties were determined (strength and range of activities). Isolated enterocin and bacteriocin-producing Lactococcus strain showed significant antimicribial activity against Listeria monocytogenes, but still the inhibition activity of Staphylococcus aurues and Escherichia coli was not detected. Based on the obtained laboratory results, in the second phase of the research, the antilisterial effect of bacteriocin isolated from Enterococcus faecalis SRB/ZS/090 and cells Lactococcus lactis ssp. lactis SRB/ZS/094 were determined, that are added as additives in the production of soft white cheese through five variants. Cheese supplemented with enterocin (E2) had the lowest aerobic mesophilic bacteria count, indicating that enterocin (E2) play an important role for bio-preservation.