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This study explores the potential probiotic properties of yeasts isolated from various Chilean honeys, focusing on Ulmo, Quillay, and Mountain honeys. Six yeast strains were identified, including Zygosaccharomyces rouxii, Candida sp., Schizosaccharomyces pombe, Rhodosporidiobolus ruineniae, Clavispora lusitaniae, and Metschnikowia chrysoperlae. Phenotypic characterization involved assessing their fermentative performance, ethanol and hops resistance, and cross-resistance. Ethanol concentration emerged as a limiting factor in their fermentative performance. The probiotic potential of these yeasts was evaluated based on resistance to high temperatures, low pH, auto-aggregation capacity, survival in simulated in vitro digestion (INFOGEST method), and antimicrobial activity against pathogens like Escherichia coli, Staphylococcus aureus, and Salmonella enteritidis. Three yeasts, Zygosaccharomyces rouxii, Schizosaccharomyces pombe, and Metschnikowia chrysoperlae, exhibited potential probiotic characteristics by maintaining cell concentrations exceeding 106 CFU/mL after in vitro digestion. They demonstrated fermentative abilities and resistance to ethanol and hops, suggesting their potential as starter cultures in beer production. Despite revealing promising probiotic and technological aspects, further research is necessary to ascertain their viability in producing fermented foods. This study underscores the innovative potential of honey as a source for new probiotic microorganisms and highlights the need for comprehensive investigations into their practical applications in the food industry.

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Non-Saccharomyces yeasts are unicellular eukaryotes that play important roles in diverse ecological niches. In recent decades, their physiological and morphological properties have been reevaluated and reassessed, demonstrating the enormous potential they possess in various fields of application. Non-Saccharomyces yeasts have gained relevance as probiotics, and in vitro and in vivo assays are very promising and offer a research niche with novel applications within the functional food and nutraceutical industry. Several beneficial effects have been described, such as antimicrobial and antioxidant activities and gastrointestinal modulation and regulation functions. In addition, several positive effects of bioactive compounds or production of specific enzymes have been reported on physical, mental and neurodegenerative diseases as well as on the organoleptic properties of the final product. Other points to highlight are the multiomics as a tool to enhance characteristics of interest within the industry; as well as microencapsulation offer a wide field of study that opens the niche of food matrices as carriers of probiotics; in turn, non-Saccharomyces yeasts offer an interesting alternative as microencapsulating cells of various compounds of interest.

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This work aimed to evaluate the performance of co-cultivation of potential probiotic yeast and lactic acid bacteria (LAB) in producing plant-based fermented beverages. The co-culture comprised LAB Lactiplantibacillus plantarum CCMA0743 with the yeasts Pichia kluyveri CCMA 0615, Pichia guilliermondii CCMA 1753 and Debaryomyces hansenii CCMA 1761 separately. The plant substrate was 75 g oat, 175 g sunflower seeds, and 75 g almonds. The viability of microorganisms in the plant-based matrix was evaluated during fermentation, storage at 4 °C, and under simulated gastrointestinal tract (GIT) conditions. Chemical analysis, antioxidant activity, and sensory profile of the beverages were also determined. The three yeasts and the LAB showed counts greater than 6.0 log CFU/mL after fermentation, and the plant-based matrix protected the yeasts during simulated digestion. P. kluyveri and D. hansenii showed higher survival than P. guilliermondii and L. plantarum after exposure to simulated GIT conditions. The pH of the plant-based matrix reduced from approximately 7 to 3.8. Lactic acid was the main organic acid produced during fermentation. In addition, 113 volatile compounds were detected by gas chromatography-mass spectrometry (GC-MS), including alcohols, aldehydes, alkanes, alkenes, acids, ester, ether, ketones, phenol, and amides. The beverage sensory profile varied with the co-culture. The co-culture D. hansenii and L. plantarum showed higher antioxidant activity than the other co-culture tested, and the homogeneous texture attribute characterized the beverage produced with this combination. Results show the suitability of tested co-cultures to produce a plant-based fermented beverage and indicate more significant potential for D. hansenii and L. plantarum co-culture as a starter for its functionalization.

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Yeast isolates from flowers and fruits from a Brazilian forest were studied. The yeasts were identified at species and strain level by PCR-RFLP and PCR-RAPD, respectively. The 46 isolated yeasts were classified into 11 different species belonging to the genera Candida, Diutina, Hanseniaspora, Meyerozyma, Pichia, Rhodotorula, and Torulaspora. A total of 20 different strains were found. In order to ascertain the probiotic potential, the resistance to gastrointestinal conditions, autoaggregation, and hydrophobicity assays were studied, along with the capacity to form biofilm. The results indicate that, although most of the strains presented better results than Saccharomyces boulardii (the only strain recognized as a probiotic yeast), four strains were the most promising, namely, Rhodotorula mucilaginosa 32, Meyerozyma caribbica 35, and Diutina rugosa 12 and 45, according to the Duncan test. Several biotechnological properties were evaluated. D. rugosa inhibited Dekkera bruxellensis. The assimilation or fermentation of seven sugars was tested, and only five of the yeasts did not show a capacity to assimilate any of the sugars under aerobic conditions. However, all strains were able to ferment at least one of the sugars under anaerobic conditions. As far as enzyme production is concerned, positive results were only found for the enzymes' amylase, pectinase, and protease. D. rugosa 42 and Hanseniaspora opuntiae 18, followed of Pichia kluyveri 26, showed high values for the production of melatonin. In conclusion, the results of this study show that several non-Saccharomyces present probiotic characteristics, and these have good potential for industrial applications in the food or biotechnology industries.

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The host microbiome plays an essential role in health and disease. Microbiome modification by pathogens or probiotics has been poorly explored especially in the case of probiotic yeasts. Next-generation sequencing currently provides the best tools for their characterization. Debaryomyces hansenii 97 (D. hansenii 97) and Yarrowia lipolytica 242 (Y. lipolytica 242) are yeasts that protect wildtype zebrafish (Danio rerio) larvae against a Vibrio anguillarum (V. anguillarum) infection, increasing their survival rate. We investigate the effect of these microorganisms on the microbiome and neutrophil response (inflammation) in zebrafish larvae line Tg(Bacmpx:GFP) i114. We postulated that preinoculation of larvae with yeasts would attenuate the intestinal neutrophil response and prevent modification of the larval microbiome induced by the pathogen. Microbiome study was performed by sequencing the V3-V4 region of the 16S rRNA gene and prediction of metabolic pathways by Piphillin in conventionally raised larvae. Survival and the neutrophil response were both evaluated in conventional and germ-free conditions. V. anguillarum infection resulted in higher neutrophil number in the intestinal area compared to non-infected larvae in both conditions. In germ-free conditions, infected larvae pre-inoculated with yeasts showed fewer neutrophil numbers than infected larvae. In both conditions, only D. hansenii 97 increased the survival of infected larvae. Beta diversity of the microbiota was modified by V. anguillarum and both yeasts, compared to non-inoculated larvae. At 3 days post-infection, V. anguillarum modified the relative abundance of 10 genera, and pre-inoculation with D. hansenii 97 and Y. lipolytica 242 prevented the modification of 5 and 6 of these genera, respectively. Both yeasts prevent the increase of Ensifer and Vogesella identified as negative predictors for larval survival (accounting for 40 and 27 of the variance, respectively). In addition, yeast pre-inoculation prevents changes in some metabolic pathways altered by V. anguillarum's infection. These results suggest that both yeasts and V. anguillarum can shape the larval microbiota configuration in the early developmental stage of D. rerio. Moreover, modulation of key taxa or metabolic pathways of the larval microbiome by yeasts can be associated with the survival of infected larvae. This study contributes to the understanding of yeast-pathogen-microbiome interactions, although further studies are needed to elucidate the mechanisms involved.

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Pineapple (Ananas comosus (L.) Merril) is a tropical fruit rich in nutrients characterized by a pleasant taste and widely consumed in several countries. It is used to produce juice, jams and wine. In this work, 150 yeasts isolates were obtained from peel (18) and spontaneously fermented pineapple pulp (132). The probiotic potential of 50 isolates was studied. Survival at pH 2.0, pepsin 3.0 g/L, and tolerance of bile salts (0.1 and 1% (w/v) were determined as indicators of survival potential of the isolates during the passage through the human gastrointestinal tract in simulated conditions. The selected isolates were also evaluated for their resistance to 6 antibiotics, antimicrobial activity against 6 pathogenic bacteria and autoaggregation and hydrophobicity properties. Five of them survived to gastrointestinal conditions, showed antibiotic resistance and autoaggregation properties. They were identified by MALDI-TOF MS and sequencing of ITS region as Candida lusitaniae (3) and Meyerozyma caribbica (2). Among these isolates, M. caribbica 9 D was evaluated in the production of a fermented pineapple beverage. The Saccharomyces cerevisiae var. boulardii was used as control, due to the fact that it is the only commercially available probiotic yeast. With M. caribbica inoculum, the beverage produced showed higher concentrations of residual glucose (24.19 g/L) and fructose (8.67 g/L), lower concentration of acetic acid (0.22 g/L); higher total phenolic compounds (196.93 mg/L), catechin (155.56 mg/L), chlorogenic acid (3.64 mg/L), vanillin (0.18 mg/L) and ferulic acid (33.2 mg/L). It was observed that M. caribbica population remained stable during refrigerated storage with cell counts greater than 7.00 log CFU/mL over 21 days. Compared to beverage produced with S. cerevisiae var. boulardii, the one produced with M. caribbica presented greater acceptance in the sensorial analysis for taste, aroma and general acceptance. The fermented pineapple beverage prepared with M. caribbica proved to be a good alternative in development of a potential probiotic beverage with different sensory and nutritional properties.

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AIMS: The aim of this work was to identify 20 yeasts isolated from autochthonal cheese starters and evaluate their technological and functional properties. METHODS AND RESULTS: The capacities of the yeasts to grow at different temperatures, pH, NaCl and lactic acid concentrations as well as the proteolytic and lipolytic activities were studied. Moreover, survival to simulated gastrointestinal digestion, hydrophobicity, antimicrobial activity against pathogens and auto- and co-aggregation abilities were evaluated. The sequentiation of a fragment from the 26S rDNA gene indicated that Kluyveromyces marxianus was the predominant species, followed by Saccharomyces cerevisiae, Clavispora lusitaniae, Kluyveromyces lactis and Galactomyces geotrichum. RAPD with primer M13 allowed a good differentiation among strains from the same species. All strains normally grew at pH 4.7-5.5 and temperatures between 15 and 35°C. Most of them tolerated 10% NaCl and 3% lactic acid. Some strains showed proteolytic (eight isolates) and/or lipolytic (four isolates) capacities. All strains evidenced high gastrointestinal resistance, moderate hydrophobicity, intermediate auto-aggregation and variable co-aggregation abilities. No strains inhibited the growth of the pathogens assayed. CONCLUSIONS: Some strains from dairy sources showed interesting functional and technological properties. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has been the first contribution to the identification and characterization of yeasts isolated from autochthonal cheese starters in Argentina. Many strains could be proposed as potential candidates to be used as probiotics and/or as co-starters in cheese productions.

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