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. 2024 Dec 17;10(12):878.
doi: 10.3390/jof10120878.

Probiotic Potential of Yeasts Isolated from Fermented Beverages: Assessment of Antagonistic Strategies Against Salmonella enterica Serovar Enteritidis

Silvia Cristina Vergara Alvarez  1   2 , María Dolores Pendón  2   3 , Ana Agustina Bengoa  2   3 , María José Leiva Alaniz  1   2 , Yolanda Paola Maturano  1   2 , Graciela Liliana Garrote  2   3
Affiliations

Probiotic Potential of Yeasts Isolated from Fermented Beverages: Assessment of Antagonistic Strategies Against Salmonella enterica Serovar Enteritidis

Silvia Cristina Vergara Alvarez et al. J Fungi (Basel). .

Abstract

Global concern about pathogenic resistance to antibiotics is prompting interest in probiotics as a strategy to prevent or inhibit infections. Fermented beverages are promising sources of probiotic yeasts. This study aimed to evaluate the antagonistic effects of Kluyveromyces marxianus, Wickerhamomyces anomalus, and Pichia manshurica strains from kefir and wine against Salmonella enterica serovar Enteritidis in intestinal epithelial cells. The ability of these yeasts to adhere to Caco-2/TC-7 cells was evaluated, as well as their influence on the ability of Salmonella to associate and invade these cells. The behavior of the pathogen was analyzed by (a) incubation of enterocytes with yeast before adding Salmonella, (b) co-incubation of Salmonella with yeast before contact with the enterocytes, and (c) incubation of Salmonella with yeast metabolites before contact with enterocytes. All yeast strains demonstrated adherence to Caco-2/TC-7 cells (33-100%) and effectively inhibited Salmonella invasion. Among the treatments, co-culture showed the greatest effect, reducing Salmonella association and invasion by more than 50%. Additionally, these yeasts modulated the epithelial immune response, significantly decreasing CCL20-driven luminescence by 60-81% (p < 0.0001). These results highlight the potential of yeasts from fermented beverages as probiotics to counteract Salmonella infections, offering a promising alternative in the fight against antibiotic resistance.

Keywords: Salmonella; antagonistic activity; kefir; non-Saccharomyces yeasts; probiotic; wine.

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Conflict of interest statement

The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Figures

Figure 1
Figure 1
Effect of yeasts strains on the association of S. Enteritidis to Caco-2/TC-7. Percentage of association of S. Enteritidis to Caco-2/TC-7 cells treated with yeasts before pathogen addition (YE), S. Enteritidis pre-cultured with yeasts (CC), and S. Enteritidis pretreated with yeasts cell-free culture supernatants (CFS). ** statistically significant differences (p ≤ 0.05) compared to the control representing 100% association.
Figure 2
Figure 2
Effect of yeast strains on the invasion of S. Enteritidis to Caco-2/TC-7. Percentage of invasion of S. Enteritidis to Caco-2/TC-7 cells treated with yeasts before pathogen addition (YE), S. Enteritidis pre-cultured with yeasts (CC), and S. Enteritidis pretreatment with yeast cell-free culture supernatants (CFS). ** statistically significant differences (p ≤ 0.05) compared to the control representing 100% invasion.
Figure 3
Figure 3
Effect of yeasts strains on the immunomodulation of Caco-2-CCL20: luc cells. Reporter cells were stimulated with flagellin (1 µg/mL) after pretreatment with yeasts. Luciferase activity was determined in a cell lysate 5 h after stimulation. Results representative from two different experiments are shown. Results are expressed as normalized luciferase activity (NAL), using the levels of stimulated cells in the absence of yeast cells as 100% of activation. **** p < 0.0001.
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