Antifungal activity of protective cultures against the yogurt drink-specific spoilage yeasts

Abstract

Fungal agents emerged as post-pasteurization contamination are responsible for the spoilage in yogurt drink. In this work, the antifungal effects of some lactic acid bacteria (LAB) on the spoilage yeasts isolated from yogurt drink (Doogh) were evaluated. First, the microbial growth in the yogurt drink samples during the storage time was investigated, and the isolated microorganisms were identified using biochemical methods and sequencing of the specific amplicons. Yeasts (3-7 log CFU ml^-1 ) were found to be the most abundant microorganisms (specific spoilage organisms) in several samples. Using the amplification technique of rDNA by ITS1 and ITS4 primers, the dominant yeasts were identified as Pichia kudriavzevii, Kluyveromyces marxianus, and Candida parapsilosis. Then, the antimicrobial activity of 37 strains of LAB against the isolated yeasts was studied using broth microdilution. Eventually, the strains of Lacticplantibacillus plantarum (245, 24, P6, and P7), Lactiplantibacillus pentosus (20), and Levilactobacillus brevis (30) exhibited significant antifungal activity. In the most effective impacts, lag times of C. parapsilosis, K. marxianus, and P. kudriavzevii were increased by almost 12-19 h, 12-19 h, and 2-6 h, respectively, while the area under the growth curve for these yeasts was reduced to lower than 40%, near 16%, and approximately 67%, in the order given. Overall, these bacteria showed high potential as the substituents for chemical preservatives in yogurt drinks. PRACTICAL APPLICATION: Spoilage yeasts were isolated from yogurt drink and identified by molecular method. Isolated yeasts belonged to Pichia, Kluyveromyces, and Candida genera. Inhibitory effects of 37 strains were evaluated against the spoilage yeasts. Cell-free supernatant was used against the isolated fungi in microdilution method. Several LAB strains showed a significant antimicrobial activity.

Keywords: LAB; antifungal activity; broth microdilution; fungal spoilage; protective culture; yogurt drink.