Amino acid profile behavior during the fermentation of Criollo cocoa beans.

The study investigated the behavior of seventeen amino acids during spontaneous (SF) and starter culture (SC) fermentation of Criollo cocoa beans from Copallín, Guadalupe and Tolopampa, Amazonas-Peru. For this purpose, liquid chromatography (UHPLC) was used to quantify amino acids. Multivariate analysis was used to differentiate the phases of the fermentation process. The percentage of essential amino acids during SC fermentation (63.4%) was higher than SF (61.8%); it was observed that the starter culture accelerated their presence and increased their concentration during the fermentation process. The multivariate analysis identified a first stage (day 0 to day 2), characterized by a low content of amino acids that increased due to protein hydrolysis. The study showed that adding the starter culture (Saccharomyces cerevisiae) to the fermentation mass increased the concentration of essential amino acids (63.0%) compared to the spontaneous process (61.8%). Moreover, this addition reduced the fermentation time (3-4 days less), demonstrating that the fermentation process with a starter culture allows obtaining a better profile of amino acids precursors of flavor and aroma.

Evaluation of stress tolerance and design of alternative culture media for the production of fermentation starter cultures in cacao.

Ecuador is one of the world's leading producers of cacao beans, and Nacional x Trinitario cacao represents one of the most distinctive varieties due to its flavor and aroma characteristics. This study aimed to evaluate the effect of the starter culture isolated from microbial diversity during the spontaneous fermentation of Nacional x Trinitario cacao. A total of 249 microbial isolates were obtained from spontaneous culture, with Lactiplantibacillus (45 %), Saccharomyces (17 %), and Acetobacter (2 %) being the most relevant genera for fermentation. Tolerance tests were conducted to select microorganisms for the starter culture. Lactiplantibacillus plantarum exhibited the highest tolerance at pH 5 and 6 % ethanol and tolerated concentrations up to 15 % for glucose and fructose. Acetobacter pasteurianus grew at pH 2 and 6 % ethanol, tolerating high sugar concentrations of up to 15 % for glucose and 30 % for fructose, with growth observed in concentrations up to 5 % for lactic and acetic acid. Subsequently, a laboratory-scale fermentation was conducted with the formulated starter culture (SC) comprising S. cerevisiae, L. plantarum, and A. pasteurianus, which exhibited high tolerance to various stress conditions. The fermentation increased alcoholic compounds, including citrusy, fruity aromas, and floral notes such as 2-heptanol and phenylethyl alcohol, respectively 1.6-fold and 5.6-fold compared to the control. Moreover, the abundance of ketones 2-heptanone and 2-nonanone increased significantly, providing sweet green herbs and fruity woody aromas. Cacao fermented with this SC significantly enhanced the favorable aroma-producing metabolites characteristic of Fine-aroma cacao. These findings underscore the potential of tailored fermentation strategies to improve cacao product quality and sensory attributes, emphasizing the importance of ongoing research in optimizing fermentation processes for the cacao industry.

Fermentation of coffee fruit with sequential inoculation of Lactiplantibacillus plantarum and Saccharomyces cerevisiae: effect on sensory attributes and chemical composition of the beans.

This study has innovative aspects related to the use of sequential inoculation technique in the coffee bean fermentation process: the inoculation of Lactiplantibacillus plantarum followed by Saccharomyces cerevisiae, in the fermentation of coffee fruit for the production of specialty natural coffees. The objective was to evaluate the effect of this technique and of the total fermentation time on the sensory attributes of the coffee beverage and on the organic acid profile, bioactive compounds, and fatty acid profile of the beans. The fermentation of coffee fruit with sequential inoculation resulted in greater acidity of the beverage and contributed to increases of up to 2 points in coffee fermented. The total fermentation time was directly related to the organic acid content, and the longer the total fermentation time was, the greater the organic acid content. The fatty acid content and bioactive compound content showed little variation among treatments.

Profile of lactic acid bacteria (MALDI-TOF-MS) and physico-chemical and microbiological characteristics of the raw milk and fresh artisanal cheese from Serra Geral, Minas Gerais, Brazil.

Artisanal cheese from Serra Geral, Minas Gerais, Brazil, stands out for its cultural asset and socio-economic relevance. However, standards of identity and quality and the peculiar terroir associated with the edaphoclimatic conditions have not been established. Therefore, the production flow diagram and the physico-chemical and microbiological quality of the raw milk, pingo (natural starter culture), production benches, water and fresh cheese were investigated for the first time. In addition, lactic acid bacteria (LAB) from cheese and its production environment were identified by MALDI-TOF. For that, 12 cheese making facilities were selected. The raw milk and pingo showed adequate physico-chemical characteristics for cheesemaking; however, high microbial counts were found. In the water, total and thermotolerant coliforms were also identified. The fresh cheeses were classified as 'high moisture and fat' and 'soft mass'. Most physico-chemical parameters were satisfactory; however, there were high counts of total coliforms, Staphylococcus spp. and coagulase-positive staphylococci. There were high counts of LAB in the raw milk, pingo, bench surface and fresh cheese. A total of 84 microbial biotypes from MRS agar were isolated. Lactococcus lactis was the predominant LAB, followed by Lactococcus garvieae. Leuconostoc mesenteroides (benches), Leuconostoc pseudomesenteroides (fresh cheese), and Enterococcus faecium (pingo) were identified sporadically. These results indicate the risks to public health associated with the consumption of the fresh cheese, and measures to improve its safety are needed.

Natural fermentation with delayed inoculation of the yeast Torulaspora delbrueckii: Impact on the chemical composition and sensory profile of natural coffee.

All coffee production stages occur in a microbiome, which is generally composed of bacteria, yeasts, and filamentous fungi. The use of starter cultures in post-harvest processing stages is an interesting alternative, since they promote faster removal of mucilage and incorporation of compounds that improve sensory quality, which can result in diverse sensory attributes for the beverage. This study was therefore developed with the objective of evaluating the effect of the following processing procedures on the chemical and sensory characteristics of the coffee beverage: first, fermentation of coffee fruit of the yellow Catucaí variety of Coffea arabica with indigenous microorganisms, followed by inoculation of the starter culture Torulaspora delbrueckii CCMA 0684 during the drying stage. The fruit was divided into two lots, which were differentiated by a natural fermentation process before drying began. The starter culture was inoculated on the coffee at different times during the drying process: at 0 h, 24 h, 48 h, or 72 h after drying began. The sensory attributes, the volatile compound composition of the roasted beans, the organic acid profile, the bioactive compounds, and the fatty acid profile of the green coffee beans were analyzed. The fatty acid and bioactive compound content showed little variation among treatments. Analysis of volatile compounds and organic acids and evaluation of sensory attributes made it possible to distinguish the two treatments. We conclude that natural fermentation of coffee fruit improve the chemical and sensory quality of the coffee beverage. The effect of natural fermentation may be before inoculation of the starter cultures or even during drying.

Effects of lactic acid bacteria and coagulase-negative staphylococci on dry-fermented sausage quality and safety: Systematic review and meta-analysis.

The meta-analysis aim was to confirm and quantifying the influence of starter cultures on microbiological and physical-chemical parameters of dry-fermented sausages at the end fermentation stage. The literature search yielded 1194 citations, and 77 studies with 178 experiments were eligible and included in the meta-analysis, a random-effects model was used to estimate the pooled weighted mean difference (MD) with 95% confidence interval (CI).The use of starter culture in dry-fermented sausages significantly reduced pH (MD: -0.364; CI: -0.414; -0.319), moisture (MD: -1.443; CI: -1.931; -0.955), aw (MD: -0.011; CI: -0.017; -0.006), Enterobacteriaceae count (MD: -1.119; CI: -1.293; -0.945), yeasts and molds count (MD: -0.351; CI: -0.691; -0.084), and increased color component a* (MD: 0.859; CI: 0.266;1.452), color component L* (MD: 1.288; CI: 0.433; 2.143), LAB count (MD: 0.981; CI: 0.696;1.267), Staphylococci count (MD: 0.484; CI: 0.293; 0.675) and TVC (MD: 0.529; CI: 0.098; 0.959). The results of the sub-analysis suggest that the addition of LAB and LAB/CNS inocula have a greater effect on the physico-chemical and microbiological parameters studied in this work. In the meta-regression analysis, a positive linear relationship was found in starter culture sausages in comparison with control batch between LAB count and the dose of starter culture added, and in the pH and Enterobacteriaceae count with the passage of fermentation days. In contrast, a negative linear relationship was found between redness and increased casing diameter of the sausages. Therefore, our work shows impact that addition of starter cultures has on safety and quality of dry-fermented sausages.

Use of microencapsulated starter cultures by spray drying in coffee under self-induced anaerobiosis fermentation (SIAF).

Using starter culture in liquid form is not economically viable in the coffee fermentation process. This work aimed to compare the fermentative performances of fresh and microencapsulated yeasts in coffee under self-induced anaerobic fermentation (SIAF). The inoculum permanence was monitored, and sugars, alcohols, acids, and volatile compounds were analyzed by chromatography. In addition, sensory analysis was performed on roasted beans. After 180 h of fermentation in the natural process, microencapsulated Torulaspora delbrueckii (MT) (7.97 × 107 cells/g) showed a higher population thanfresh Torulaspora delbrueckii (FT) (1.76 × 107 cells/g). The same acids and volatile compounds were detected in coffees with fresh and microencapsulated yeast. However, the yeast state influenced the concentration of the compounds. In pulped coffee, the coffee inoculated withmicroencapsulated Saccharomyces cerevisiae (MS) obtained the highest concentration of alcohols, esters, pyrazines, and others compared with fresh Saccharomyces cerevisiae (FS), with an increase of up to 47%. Furthermore, the coffee inoculated with MT obtained the highest concentration in almost all chemical classes in both processes compared with FT. These differences ranged up to 55%. Regarding sensory analysis, coffees inoculated with MS showed dominant notes of fruity, caramel, and nuts in the natural process. Otherwise, in pulped process, coffees inoculated with MT showed caramel, honey, and nuts. Therefore, the microencapsulated yeasts were metabolically active and may be considered with commercial potential. Considering the parameters analyzed, the most suitable yeast for natural and pulped processing would be MS and MT, respectively.

High Level of Interaction between Phages and Bacteria in an Artisanal Raw Milk Cheese Microbial Community.

Microbial starter cultures are used in the production of many cheeses around the world, such as Parmigiano-Reggiano, in Italy, Époisses, in France, and Canastra, in Brazil, providing many of the unique features of these cheeses. Bacteriophages (phages) are ubiquitous and well known to modulate the structure of bacterial communities, and recent data indicate that cheeses contain a high abundance of naturally occurring phages. Here, we analyze the viral and bacterial metagenomes of Canastra cheese: a traditional artisanal Brazilian cheese produced using an endogenous starter culture and raw milk. Over 1,200 viral operational taxonomic units were recovered using both isolated viral-like particles and complete metagenomic DNA. Common viral families identified included Siphoviridae and Myoviridae, with 40% of putative phage genomes unidentified at the family level of classification. We observed very high phage diversity, which varied greatly across different cheese producers, with 28% of phage genomes detected in only one producer. Several metagenome-assembled genomes were recovered for lactic acid-producing bacteria, as well as nonstarter bacterial species, and we identified several phage-bacterium interactions, at the strain level of resolution, varying across distinct cheese producers. We postulate that at least one bacterial strain detected could be endogenous and unique to the Canastra cheese-producing region in Brazil and that its growth seems to be modulated by autochthonous phages present in this artisanal production system. This phage-host relationship is likely to influence the fermentation dynamics and ultimately the sensorial profile of these cheeses, with implications for other similar cheese production systems around the world. IMPORTANCE Our work demonstrated a dynamic yet stable microbial ecosystem during cheese production using an endogenous starter culture. This was observed across several distinct producers and was marked by genomic evidence of continued phage-bacterium interactions, such as the presence of bacterial defense mechanisms. Furthermore, we provide evidence of unique microbial signatures for each individual cheese producer studied in the region, a fact that may have profound consequences on product traceability. This was the first effort to describe and understand the bacteriophage composition and ecological dynamics within the Brazilian Canastra cheese production system. The study of this prototypical backslopping production system provides a solid background for further mechanistic studies of the production of many cheeses around the world.

Traditional Fermented Foods from Ecuador: A Review with a Focus on Microbial Diversity.

The development of early civilizations was greatly associated with populations' ability to exploit natural resources. The development of methods for food preservation was one of the pillars for the economy of early societies. In Ecuador, food fermentation significantly contributed to social advances and fermented foods were considered exclusive to the elite or for religious ceremonies. With the advancement of the scientific research on bioprocesses, together with the implementation of novel sequencing tools for the accurate identification of microorganisms, potential health benefits and the formation of flavor and aroma compounds in fermented foods are progressively being described. This review focuses on describing traditional fermented foods from Ecuador, including cacao and coffee as well as less popular fermented foods. It is important to provide new knowledge associated with nutritional and health benefits of the traditional fermented foods.

Microencapsulation of epiphytic coffee yeasts by spray drying using different wall materials: Implementation in coffee medium.

The storage of microorganisms in liquid form is the main drawback of commercializing epiphytic coffee yeasts. This work aimed to evaluate the fermentative performance of microencapsulated yeasts by spray drying in a coffee peel and pulp media (CPM). The yeasts, Saccharomyces cerevisiae CCMA 0543, Torulaspora delbrueckii CCMA 0684, and Meyerozyma caribbica CCMA 1738, were microencapsulated using maltodextrin DE10 (MD), high maltose (MA), and whey powder (WP) as wall materials. A Central Composite Rotational Design (CCRD) was used to investigate the effect of operating parameters on the microcapsules' cell viability, drying yield, and water activity. Yeasts reached cell viability and drying yields above 90 and 50 %, respectively. WP maintained the cell viability of the three yeasts over 90 days of storage at room temperature (25 °C) and was selected as a wall material for the three yeasts. M. caribbica showed to be more sensitive to spray drying and less resistant to storage. Some differences were found in the fermentation of the CPM medium, but the microencapsulated yeasts maintained their biotechnological characteristics. Therefore, the microencapsulation of epiphytic coffee yeasts by spray drying was promising to be used in the coffee fermentation process.

Microencapsulation by spray drying of coffee epiphytic yeasts Saccharomyces cerevisiae CCMA 0543 and Torulaspora delbrueckii CCMA 0684.

The objective of this work was to evaluate the microencapsulation feasibility of Saccharomyces cerevisiae CCMA 0543 and Torulaspora delbrueckii CCMA 0684 in three different compositions of wall material by spray-dryer. The yeasts (109 CFU mL-1) were microencapsulated separately using maltodextrin (15%), maltodextrin (15%) with sucrose (2%), or maltose (2%) as wall material. The viability was evaluated for 6 months at two different temperatures (7 and 25 °C). The yield, cell viability after spray drying, and characterization of the microcapsules were performed. Results indicate that cell viability ranged between 94.06 and 97.97%. After 6 months, both yeasts stored at 7 °C and 25 °C presented 107 and 102 CFU mL-1, respectively. Regarding Fourier-transform infrared spectroscopy analysis, all microencapsulated yeasts presented typical spectra footprints of maltodextrin. After 6 months of storage, S. cerevisiae CCMA 0543 obtained a 10.8% increase in cell viability using maltodextrin with maltose as wall material compared to maltodextrin and maltodextrin with sucrose. However, T. delbrueckii CCMA 0684 obtained a 13.5% increase in cell viability using only maltodextrin. The study showed that maltodextrin as a wall material was efficient in the microencapsulation of yeasts. It is possible to assume that maltose incorporation increased the cell viability of S. cerevisiae CCMA 0543 during storage.

Effect of ultrasound application on the growth of S. xylosus inoculated in by-products from the poultry industry.

A wide variety of by-products are produced by the industry when animals are slaughtered. However, the proteins present in these by-products, are not being fully useable, in the elaboration of value-added products. Staphylococcus xylosus is commonly used as a starter culture in meat products subjected to ripening for a long period, as it produces proteolytic and lipolytic enzymes that improve the sensory quality of the products. Ultrasound (US) has been arousing interest in the meat industry, as it reduces processing time and also improves the technological and sensory quality of meat products. However, the stimulate effect of US on the growth of S. xylosus in by-products from the poultry industry is still unknown. Thus, this study aimed to evaluate the stimulate effect of US on the growth of S. xylosus inoculated in by-products from the poultry industry. S. xylosus was inoculated (5.63 log CFU/g) in sterilized by-products from the poultry, which were then sonicated at 37 °C for 0, 15, 30, and 45 min according to the following parameters: frequencies of 130 and 35 kHz, amplitudes of 50% and 80% and normal and degas operating modes. The sonicated samples were incubated at 37 °C for 0, 24, 48, and 72 h. Soon after sonication, no stimulate effect of US was observed on the growth of S. xylosus. However, after 24 h of incubation, the samples sonicated for 15 and 30 min in normal mode, at 35 and 130 kHz, and amplitudes of 50 and 80% exhibited better stimulate effect at the growth S. xylosus counts (p < 0.01) when compared to the Control, with values of 8.23 and 7.77 log CFU/g, respectively. These results can be exploited to obtain new added-value products, having as raw material by-products from the poultry industry.

Biochemical, microbiological and technological characteristics of type II sourdoughs produced with lactic acid bacteria single-strains / Características bioquímicas, microbiológicas e tecnológicas de sourdoughs tipo II produzidos com uma única cepa de bactéria ácido lática

The interplay between biochemical characteristics and the generation of volatile compounds in 11 type II sourdough fermented by single strains of lactic acid bacteria (LAB) was studied. Samples were collected at 0, 6, 9, 12, 15, 18 and 24h for analyses of microbial growth, pH, titratable acidity and CO2 production. During the first 12h, the LABs entered the stationary phase, and the formation of organic and carboxyl acids, alcohols, and esters were observed. Although acidity is an important characteristic of sourdough, in this work increasing the acetic acid content decreased yeast growth and the CO2 retention capacity of the doughs. The main carbohydrate consumed by autochthonous yeast was influenced by the LAB added (homo-or heterofermentative), as observed by correlation analysis. Maltose and glucose showed a strong and negative correlation with the yeast cell density in the dough fermented by homo and heterofermentative LAB, respectively. Moreover, LAB had an important effect on the aromatic profile, being the alcohols, aldehydes, alkanes, organics acids and esters mainly groups characterized. Altogether, 100 different volatile compounds were identified; however, each dough had a different volatile profile. This study shows, for the first time, the influence of a single strain of LAB on the characteristics of type II sourdough.(AU)

As características bioquímicas e a produção de compostos voláteis em 11 diferentes sourdough tipo II produzido com uma única cepa de bactérias do ácido láctico (BAL) em foi estudada. As amostras foram coletadas às 0, 6, 9, 12, 15, 18 e 24 h para análises de crescimento microbiano, pH, acidez titulável e produção de CO2. Durante as primeiras 12 h, as BAL entraram em fase estacionária, sendo observada a formação de ácidos orgânicos e carboxílicos, álcoois e ésteres. Embora a acidez seja uma característica importante do sourdough, neste trabalho o aumento do teor de ácido acético diminuiu o crescimento das leveduras e a capacidade de retenção de CO2 nas massas. Também foi observado que o principal carboidrato consumido pelas leveduras autóctones foi influenciado pela BAL adicionada (homo ou heterofermentativas), conforme observado pela análise de correlação. A maltose e a glicose apresentaram uma correlação forte e negativa com a densidade celular de levedura na massa fermentada por BAL homo e heterofermentativas, respectivamente. Além disso, a BAL teve efeito importante no perfil aromático, sendo os álcoois, aldeídos, alcanos, ácidos orgânicos e ésteres os principais compostos caracterizados. Ao todo, foram identificados 100 compostos voláteis diferentes; no entanto, cada massa apresentou um perfil volátil diferente. Este estudo mostra, pela primeira vez, a influência de uma única cepa de BAL nas características de sourdough tipo II.(AU)

Probiotic Properties, Prebiotic Fermentability, and GABA-Producing Capacity of Microorganisms Isolated from Mexican Milk Kefir Grains: A Clustering Evaluation for Functional Dairy Food Applications.

Isolation and functional characterization of microorganisms are relevant steps for generating starter cultures with functional properties, and more recently, those related to improving mental health. Milk kefir grains have been recently investigated as a source of health-related strains. This study focused on the evaluation of microorganisms from artisanal Mexican milk kefir grains regarding probiotic properties, in vitro fermentability with commercial prebiotics (lactulose, inulin, and citrus pectin), and γ-aminobutyric acid (GABA)-producing capacity. Microorganisms were identified belonging to genera Lactococcus, Lactobacillus, Leuconostoc, and Kluyveromyces. The probiotic properties were assessed by aggregation abilities, antimicrobial activity, antibiotic susceptibility, and resistance to in vitro gastrointestinal digestion, showing a good performance compared with commercial probiotics. Most of isolates maintained a concentration above 6 log colony forming units/mL after the intestinal phase. Specific isolates of Kluyveromyces (BIOTEC009 and BIOTEC010), Leuconostoc (BIOTEC011 and BIOTEC012), and Lactobacillus (BIOTEC014 and BIOTEC15) showed a high fermentability in media supplemented with commercial prebiotics. The capacity to produce GABA was classified as medium for L. lactis BIOTEC006, BIOTEC007, and BIOTEC008; K. lactis BIOTEC009; L. pseudomesenteroides BIOTEC012; and L. kefiri BIOTEC014, and comparable to that obtained for commercial probiotics. Finally, a multivariate approach was performed, allowing the grouping of 2-5 clusters of microorganisms that could be further considered new promising cultures for functional dairy food applications.

Criteria for lactic acid bacteria screening to enhance silage quality.

The main challenge of ensiling is conserving the feed through a fermentative process that results in high nutritional and microbiological quality while minimizing fermentative losses. This challenge is of growing interest to farmers, industry and research and involves the use of additives to improve the fermentation process and preserve the ensiled material. Most studies involved microbial additives; lactic acid bacteria (LAB) have been the focus of much research and have been widely used. Currently, LABs are used in modern and sustainable agriculture because of their considerable potential for enhancing human and animal health. Although the number of studies evaluating LABs in silages has increased, the potential use of these micro-organisms in association with silage has not been adequately studied. Fermentation processes using the same strain produce very different results depending on the unique characteristics of the substrate, so the choice of silage inoculant for different starting substrates is of extreme importance to maximize the nutritional quality of the final product. This review describes the current scenario of the bioprospecting and selection process for choosing the best LAB strain as an inoculant for ensiling. In addition, we analyse developments in the fermentation process and strategies and methods that will assist future studies on the selection of new strains of LAB as a starter culture or inoculant.

Soymilk fermentation: effect of cooling protocol on cell viability during storage and in vitro gastrointestinal stress.

This work covers soymilk fermentation by starter and probiotic cultures and explores the influence of cooling protocol on cell viability, organic acid production, sugar consumption, fatty acid profile, and cell survival to in vitro gastrointestinal stress. After fermentation at 37 °C by mono- or co-cultures of Streptococcus thermophilus (St), Lactobacillus bulgaricus (Lb), and Lactobacillus paracasei (Lp), fermented soymilk was cooled directly at 4 °C for 28 days or cooled in two phases (TPC), i.e., by preceding that step by another at 25 °C for 8 h. Soybean milk fermentation by Lb alone lasted longer (15 h) than by StLb or StLbLp (9 h). In ternary culture, TPC increased Lp viability, linoleic, and lactic acid concentrations by 3.8, 22.6, and 96.2%, respectively, whereas the cooling protocol did not influence Lp and St counts after in vitro gastrointestinal stress. Graphical abstract.

Influence of fermentation conditions on the sensorial quality of coffee inoculated with yeast.

Fermentation is one of the post-harvest steps that influence coffee quality. This work evaluated the effect of Saccharomyces cerevisiae (CCMA 0543) and Torulaspora delbrueckii (CCMA 0684) inoculation on the quality of natural and pulped natural processed coffee in different producing regions. Yeast populations were assessed by the real-time polymerase chain reaction. Volatile and nonvolatile compounds were evaluated by gas chromatography-mass spectrometry and high-performance liquid chromatography, respectively. S. cerevisiae was predominant during spontaneous (average of 4 log cells/g) and inoculated (average of 7 log cells/g) fermentations in both processes. T. delbrueckii showed a similar population (3.79 log cells/g) in all assays. Glucose and fructose were the most detected sugars in coffee beans. Succinic acid was found at the end of the fermentative process. The lactic acid concentration was inversely proportional to ethanol concentration. Pyrrole and furan, which are volatile groups, allowed to differentiate the coffee processing methods. Yeast inoculation modified the sensorial profile and increased the coffee beverage scores by up to 5 points. S. cerevisiae inoculation was most suitable for pulped natural coffee, and T. delbrueckii inoculation showed the best performance in natural coffee.

Fungal Diversity Analysis of Grape Musts from Central Valley-Chile and Characterization of Potential New Starter Cultures.

Autochthonous microorganisms are an important source of the distinctive metabolites that influence the chemical profile of wine. However, little is known about the diversity of fungal communities associated with grape musts, even though they are the source of local yeast strains with potential capacities to become starters during fermentation. By using internal transcribed spacer (ITS) amplicon sequencing, we identified the taxonomic structure of the yeast community in unfermented and fermented musts of a typical Vitis vinifera L. var. Sauvignon blanc from the Central Valley of Chile throughout two consecutive seasons of production. Unsurprisingly, Saccharomyces represented the most abundant fungal genus in unfermented and fermented musts, mainly due to the contribution of S. uvarum (42.7%) and S. cerevisiae (80%). Unfermented musts were highly variable between seasons and showed higher values of fungal diversity than fermented musts. Since microbial physiological characterization is primarily achieved in culture, we isolated nine species belonging to six genera of fungi from the unfermented must samples. All isolates were characterized for their potential capacities to be used as new starters in wine. Remarkably, only Metschnikowia pulcherrima could co-exist with a commercial Saccharomyces cerevisiae strain under fermentative conditions, representing a feasible candidate strain for wine production.

Biotransformation of Selenium by Lactic Acid Bacteria: Formation of Seleno-Nanoparticles and Seleno-Amino Acids.

Selenium (Se) is an essential micronutrient for the majority of living organisms, and it has been identified as selenocysteine in the active site of several selenoproteins such as glutathione peroxidase, thioredoxin reductase, and deiodinases. Se deficiency in humans is associated with viral infections, thyroid dysfunction, different types of cancer, and aging. In several European countries as well as in Argentina, Se intake is below the recommended dietary Intake (RDI). Some lactic acid bacteria (LAB) can accumulate and bio-transform selenite (toxic) into Se-nanoparticles (SeNPs) and Se-amino acids (non-toxic). The microbial growth, Se metabolite distribution, and the glutathione reductase (involved in selenite reduction) activity of Se-enriched LAB were studied in this work. The ninety-six assayed strains, belonging to the genera Lactococcus, Weissella, Leuconostoc, Lactobacillus, Enterococcus, and Fructobacillus could grow in the presence of 5 ppm sodium selenite. From the total, eight strains could remove more than 80% of the added Se from the culture medium. These bacteria accumulated intracellularly between 1.2 and 2.5 ppm of the added Se, from which F. tropaeoli CRL 2034 contained the highest intracellular amount. These strains produced only the seleno-amino acid SeCys as observed by LC-ICP-MS and confirmed by LC-ESI-MS/MS. The intracellular SeCys concentrations were between 0.015 and 0.880 ppm; Lb. brevis CRL 2051 (0.873 ppm), Lb. plantarum CRL 2030 (0.867 ppm), and F. tropaeoli CRL 2034 (0.625 ppm) were the strains that showed the highest concentrations. Glutathione reductase activity values were higher when the strains were grown in the presence of Se except for the F. tropaeoli CRL 2034 strain, which showed an opposite behavior. The cellular morphology of the strains was not affected by the presence of Se in the culture medium; interestingly, all the strains were able to form spherical SeNPs as determined by transmission electron microscopy (TEM). Only two Enterococcus strains produced the volatile Se compounds dimethyl-diselenide identified by GC-MS. Our results show that Lb. brevis CRL 2051, Lb. plantarum CRL 2030, and F. tropaeoli CRL 2034 could be used for the development of nutraceuticals or as starter cultures for the bio-enrichment of fermented fruit beverages with SeCys and SeNPs.

Probiotic Potential and Technological Properties of Bacteriocinogenic Lactococcus lactis Subsp. Lactis UTNGt28 from a Native Amazonian Fruit as a Yogurt Starter Culture.

A native Lactococcus lactis subsp. lactis UTNGt28 (GenBank accession no: MG675576.1) isolated from Amazonian fruit of the tropical Caimitillo (Chrysophyllum oliviforme) tree and the commercial strain Lactococcus lactis subsp lactis ATCC11454 (LacAT) were targeted ex vitro in whole milk in combination with Streptococcus thermophilus ATCC19258 to obtain a fermented probiotic beverage. Concomitant with cell viability determination during storage (28 days), the pH, titratable acidity, syneresis, protein and fat were evaluated. The results indicated that neither UTNGt28 nor LacAT displayed a high capacity to ferment whole milk and survive during storage; a statistically significant difference (p < 0.05) in cell viability was registered for UTNGt28 compared with LacAT when inoculated alone or in combination with S. thermophilus. A principal component analysis showed a clear difference between the yogurt formulations at day 1 and 28 of storage. The PC 1 explained 46.8% of the total variance (day 28), was loaded in the negative (-) direction with titratable acidity (% lactic acid), while the PC 2 explained 22.5% (day 1) with pH. PC 1 was loaded in the positive (+) direction with pH, cell viability, syneresis, fat and protein. Overall results indicated that UTNGt28 has the technological properties for further development of a new probiotic product.