Production of flavor-active compounds and physiological impacts in immobilized Saccharomyces spp. cells during beer fermentation.

Yeast immobilization in beer fermentation has recently regained attention, due to the expansion of the craft beer market and the diversification of styles and flavors. The aim of this study was to evaluate the physiological differences between immobilized and free yeast cells with a focus on flavor-active compounds formation. Three strains of Saccharomyces spp. (SY025, SY067, SY001) were evaluated in both free and immobilized (using a cellulose-based support, referred as ImoYeast) forms during static batch fermentations of 12 °P malt extract. Immobilized cells showed higher glycerol (SY025, 40%; SY067, 53%; SY001, 19%) and biomass (SY025, 67%; SY067, 78%; SY001, 56%) yields than free cells. Conversely, free cells presented higher ethanol yield (SY025, 9%; SY067, 9%; SY001, 13%). Flavor-active compounds production exhibited significant alterations between immobilized and free cells systems, for all strains tested. Finally, a central composite design with varying initial biomass (X0) and substrate (S0) concentrations was conducted using strain SY025, which can be helpful to modulate the formation of one or more flavor-active compounds. In conclusion, yeast immobilization in the evaluated support resulted in flavor alterations that can be exploited to produce different beer styles.

An integrative taxonomy approach reveals Saccharomyces chiloensis sp. nov. as a newly discovered species from Coastal Patagonia.

Species delineation in microorganisms is challenging due to the limited markers available for accurate species assignment. Here, we applied an integrative taxonomy approach, combining extensive sampling, whole-genome sequence-based classification, phenotypic profiling, and assessment of interspecific reproductive isolation. Our work reveals the presence of a distinct Saccharomyces lineage in Nothofagus forests of coastal Patagonia. This lineage, designated Saccharomyces chiloensis sp. nov., exhibits 7% genetic divergence from its sister species S. uvarum, as revealed by whole-genome sequencing and population analyses. The South America-C (SA-C) coastal Patagonia population forms a unique clade closely related to a previously described divergent S. uvarum population from Oceania (AUS, found in Australia and New Zealand). Our species reclassification is supported by a low Ortho Average Nucleotide Identity (OANI) of 93% in SA-C and AUS relative to S. uvarum, which falls below the suggested species delineation threshold of 95%, indicating an independent evolutionary lineage. Hybrid spore viability assessment provided compelling evidence that SA-C and AUS are reproductively isolated from S. uvarum. In addition, we found unique structural variants between S. chiloensis sp. nov. lineages, including large-scale chromosomal translocations and inversions, together with a distinct phenotypic profile, emphasizing their intraspecies genetic distinctiveness. We suggest that S. chiloensis sp. nov diverged from S. uvarum in allopatry due to glaciation, followed by post-glacial dispersal, resulting in distinct lineages on opposite sides of the Pacific Ocean. The discovery of S. chiloensis sp. nov. illustrates the uniqueness of Patagonia's coastal biodiversity and underscores the importance of adopting an integrative taxonomic approach in species delineation to unveil cryptic microbial species. The holotype of S. chiloensis sp. nov. is CBS 18620T.

Expression patterns of Mal genes and association with differential maltose and maltotriose transport rate of two Saccharomyces pastorianus yeasts.

Beer brewing is a well-known process that still faces great challenges, such as the total consumption of sugars present in the fermentation media. Lager-style beer, a major worldwide beer type, is elaborated by Saccharomyces pastorianus (Sp) yeast, which must ferment high maltotriose content worts, but its consumption represents a notable problem, especially among Sp strains belonging to group I. Factors, such as fermentation conditions, presence of maltotriose transporters, transporter copy number variation, and genetic regulation variations contribute to this issue. We assess the factors affecting fermentation in two Sp yeast strains: SpIB1, with limited maltotriose uptake, and SpIB2, known for efficient maltotriose transport. Here, SpIB2 transported significantly more maltose (28%) and maltotriose (32%) compared with SpIB1. Furthermore, SpIB2 expressed all MAL transporters (ScMALx1, SeMALx1, ScAGT1, SeAGT1, MTT1, and MPHx) on the first day of fermentation, whereas SpIB1 only exhibited ScMalx1, ScAGT1, and MPH2/3 genes. Some SpIB2 transporters had polymorphic transmembrane domains (TMD) resembling MTT1, accompanied by higher expression of these transporters and its positive regulator genes, such as MAL63. These findings suggest that, in addition to the factors mentioned above, positive regulators of Mal transporters contribute significantly to phenotypic diversity in maltose and maltotriose consumption among the studied lager yeast strains.IMPORTANCEBeer, the third most popular beverage globally with a 90% market share in the alcoholic beverage industry, relies on Saccharomyces pastorianus (Sp) strains for lager beer production. These strains exhibit phenotypic diversity in maltotriose consumption, a crucial process for the acceptable organoleptic profile in lager beer. This diversity ranges from Sp group II strains with a notable maltotriose-consuming ability to Sp group I strains with limited capacity. Our study highlights that differential gene expression of maltose and maltotriose transporters and its upstream trans-elements, such as MAL gene-positive regulators, adds complexity to this variation. This insight can contribute to a more comprehensive analysis needed to the development of controlled and efficient biotechnological processes in the beer brewing industry.

Wild Patagonian yeast improve the evolutionary potential of novel interspecific hybrid strains for lager brewing.

Lager yeasts are limited to a few strains worldwide, imposing restrictions on flavour and aroma diversity and hindering our understanding of the complex evolutionary mechanisms during yeast domestication. The recent finding of diverse S. eubayanus lineages from Patagonia offers potential for generating new lager yeasts with different flavour profiles. Here, we leverage the natural genetic diversity of S. eubayanus and expand the lager yeast repertoire by including three distinct Patagonian S. eubayanus lineages. We used experimental evolution and selection on desirable traits to enhance the fermentation profiles of novel S. cerevisiae x S. eubayanus hybrids. Our analyses reveal an intricate interplay of pre-existing diversity, selection on species-specific mitochondria, de-novo mutations, and gene copy variations in sugar metabolism genes, resulting in high ethanol production and unique aroma profiles. Hybrids with S. eubayanus mitochondria exhibited greater evolutionary potential and superior fitness post-evolution, analogous to commercial lager hybrids. Using genome-wide screens of the parental subgenomes, we identified genetic changes in IRA2, IMA1, and MALX genes that influence maltose metabolism, and increase glycolytic flux and sugar consumption in the evolved hybrids. Functional validation and transcriptome analyses confirmed increased maltose-related gene expression, influencing greater maltotriose consumption in evolved hybrids. This study demonstrates the potential for generating industrially viable lager yeast hybrids from wild Patagonian strains. Our hybridization, evolution, and mitochondrial selection approach produced hybrids with high fermentation capacity and expands lager beer brewing options.

Addition of Saccharomyces eubayanus to SCOBY fermentations modulates the chemical and volatile compound profiles in kombucha.

Kombucha is a fermented beverage derived from a sweetened tea fermentation inoculated with a bacteria-yeast consortium referred to as Symbiotic Culture of Bacteria and Yeast (SCOBY). Different SCOBY cultures can impact the beverage's quality and make the whole process highly variable. Adding Saccharomyces yeast cultures to the fermentation process can avoid stalled fermentations, providing a reproducible beverage. Here, we explored using different Saccharomyces eubayanus strains together with SCOBY in the context of kombucha fermentation. Our results show that yeast x SCOBY co-cultures exhibited a robust fermentation profile, providing ethanol and acetic acid levels ranging from 0,18-1,81 %v/v and 0,35-1,15 g/L, respectively. The kombucha volatile compound profile of co-cultures was unique, where compounds such as Isopentyl acetate where only found in yeast x SCOBY fermentations. Metabarcoding revealed that the SCOBY composition was also dependent on the S. eubayanus genotype, where besides Saccharomyces, amplicon sequence variants belonging to Brettanomyces and Starmerella were detected. These differences concomitated global changes in transcript levels in S. eubayanus related to the metabolism of organic molecules used in kombucha fermentation. This study highlights the potential for exploring different S. eubayanus strains for kombucha fermentation, and the significant yeast genotype effect in the profile differentiation in this process.

Effectiveness of Saccharomyces Boulardii CNCM I-745 probiotic in acute inflammatory viral diarrhoea in adults: results from a single-centre randomized trial.

BACKGROUND: Probiotics are effective for treating acute infectious diarrhoea caused by bacteria, but there are inconsistent results for the effectiveness of probiotics for diarrhoea caused by viruses. In this article we want to determine whether Sb supplementation has an effect on acute inflammatory viral diarrhoea diagnosed with the multiplex panel PCR test. The aim of this study was to evaluate the efficacy of Saccharomyces boulardii (Sb) as a treatment in patients diagnosed with viral acute diarrhoea. METHODS: From February 2021 to December 2021, 46 patients with a confirmed diagnosis of viral acute diarrhoea diagnosed with the polymerase chain reaction multiplex assay were enrolled in a double-blind, randomized placebo-controlled trial. Patients received paracetamol 500 mg as a standard analgesic and 200 mg of Trimebutine as an antispasmodic treatment plus 600 mg of Sb (n = 23, 1 × 109/100 mL Colony forming unit) or a placebo (n = 23) orally once daily for eight days. The improvement in and severity of symptoms were measured using a symptom diary, the Patient Global Impression and the Patient Global Impression of Change scales (days 4 and 8), both answered and recorded by the patient. RESULTS: Of the 46 patients who completed treatment, 24 (52%) were men and 22 (48%) were women. The average age was 35.6 ± 12.28 years (range 18 to 61 years). The average duration of the evolution of illness at the time of diagnosis was 0.85 ± 0.73 days (maximum 2 days). On day 4 after the diagnosis, 20% reported pain and 2% reported fever, but on day 8, no patient reported pain or fever. On day 4, 70% of patients in the Sb group and 26% in the placebo group reported improvement (P = 0.03), based on the Patients' Global Impression of Change scale, which assesses patient's rating of overall improvement. These findings suggest that 3 to 4 days of treatment with Sb helped to improve symptoms of diarrhoea caused by a virus. CONCLUSION: Treatment with Sb on acute inflammatory diarrhoea of viral aetiology shows no changes regarding the severity of the symptoms; nevertheless, it seems to impact improvement positively. TRIAL REGISTRATION: 22CEI00320171130 dated on 16/12/2020, NCT05226052 dated on 07/02/2022.

Sugarcane yeast (Saccharomyces cerevisiae) and ß-mannanase enzyme in broilers' diets / Levedura de cana-de-açúcar (Saccharomyces cerevisiae) e enzima ß-mananase em dietas para frangos de corte

Our objective was to evaluate the effect of supplementing ß-mannanase enzyme, with and without sugarcane yeast (Saccharomyces cerevisiae), on broiler chickens aged 1-21 days. The study used 720 one-day-old male Cobb chicks in a randomized design, with six treatments and six replications of 20 birds each. The treatments were: basal diet (BD), BD + ß-mannanase (100 g/t), 7% sugarcane yeast (SY), SY + ß-mannanase (80 g/t), SY + ß-mannanase (100 g/t), and SY + ß-mannanase (120 g/t). Zootechnical results (animal performance) were evaluated during the pre-starter (1 to 7 day-old) and starter (1 to 21 day-old) phases, as well as small intestine morphometry (duodenum, jejunum, and ileum) and diets' economic viability. Data were subjected to variance analysis using the SAS software, and the means were compared by the Student Newman Keuls (SNK) test. In the pre-starter phase, treatments with YD + ß-mannanase (80, 100, and 120 g/t) showed the best feed conversion averages. In the starter phase, chickens consuming the basal diet (BD) and BD + ß-mannanase showed better average weights, weight gains, and feed conversion rates. For intestinal morphometry, shallower ileal crypts were observed in the treatment with YD + ß-mannanase (120 g/t) compared to the basal diet, and wider ileal villi were observed in the treatment with YD + ß-mannanase (100 g/t) compared to the diet with YD + ß-mannanase (80 g/t). The thickness of the muscular wall in the duodenum was lower in chickens consuming BD compared to BD + ß-mannanase (100 g/t), higher in YD and supplementation with 100 g/t compared to 80 and 120 g/t in the jejunum, and higher in diets with ß-mannanase supplementation compared to BD and YD in the ileum. For economic viability, adding 7% sugar cane yeast, with or without enzyme, increased the average feed cost and cost index, and reduced the economic efficiency index. Based on the zootechnical results, YD + ß-mannanase (120 g/t) is recommended for the pre-starter and starter phases. However, using sugar cane yeast with or without ß-mannanase enzyme supplementation is not economically viable in the 1- to 21-day period.

Objetivou-se avaliar o efeito da suplementação da enzima ß-mananase com e sem levedura de cana de açúcar (Saccharomyces cerevisiae) para frangos de corte 1 a 21 dias de idade. Utilizou-se 720 pintos de corte de um dia de idade, machos, da linhagem Cobb, distribuídos em delineamento inteiramente casualizado com seis tratamentos e, seis repetições de 20 aves cada. Os tratamentos foram: Dieta basal (DB); DB + ß-mananase (100 g/t); DB + 7% de levedura de cana-de-açúcar (DL); DL + ß-mananase (80 g/t); DL + ß-mananase (100 g/t); DL + ß-mananase (120 g/t). Avaliou-se o desempenho zootécnico na fase pré-inicial (1 a 7) e na fase inicial (1 a 21 dias de idade), a morfometria do intestino delgado (duodeno, jejuno e íleo) e, a viabilidade econômica da ração. Os dados foram submetidos à análise da variância do programa SAS e as médias foram comparadas pelo teste de Student Newmann Keuls (SNK). Na fase pré-inicial, os tratamentos com DL + ß-mananase (80, 100 e 120 g/t) apresentaram as melhores médias de conversão alimentar. Na fase inicial, aos frangos que consumiram à dieta basal (DB) e DB + ß-mananase apresentaram peso médio, ganho de peso e conversão alimentar melhores. Para a morfometria intestinal, criptas ileais mais rasas foram observadas no tratamento com DL+ ß-mananase (120 g/t) em relação a dieta basal e vilos ileais mais largos foram observados no tratamento com DL+ ß-mananase (100 g/t) em relação a dieta com DL+ ß-mananase (80 g/t). A espessura da parede muscular, no duodeno, foi menor nos frangos que consumiram a DB em relação a DB + ß-mananase (100 g/t), no jejuno, foi maior na DL e na suplementação com 100 g/t em relação a 80 e 120 g/t e no íleo, foi maior nas dietas com suplementação de ß-mananase em relação a DB e DL. Para a viabilidade econômica, a adição de 7% de levedura de cana-de-açúcar, com ou sem enzima proporcionaram aumento do custo médio de ração e índice de custo, e redução do índice de eficiência econômica. Com base nos resultados zootécnicos recomenda-se DL + ß-mananase (120g/t) para a fase pré-inicial e inicial. Contudo, o uso de levedura de cana-de-açúcar com e sem suplementação da enzima ß-mananase não é economicamente viável no período de 1 a 21 dias.

Extracellular expression of Saccharomyces cerevisiae's L-asparaginase II in Pichia pastoris results in novel enzyme with better parameters.

L-asparaginase (ASNase) is an efficient inhibitor of tumor development, used in chemotherapy sessions against acute lymphoblastic leukemia (ALL) tumor cells; its use results in 80% complete remission of the disease in treated patients. Saccharomyces cerevisiae's L-asparaginase II (ScASNaseII) has a high potential to substitute bacteria ASNase in patients that developed hypersensitivity, but the endogenous production of it results in hypermannosylated immunogenic enzyme. Here we describe the genetic process to acquire the ScASNaseII expressed in the extracellular medium. Our strategy involved a fusion of mature sequence of protein codified by ASP3 (amino acids 26-362) with the secretion signal sequence of Pichia pastoris acid phosphatase enzyme; in addition, this DNA construction was integrated in P. pastoris Glycoswitch® strain genome, which has the cellular machinery to express and secrete high quantity of enzymes with humanized glycosylation. Our data show that the DNA construction and strain employed can express extracellular asparaginase with specific activity of 218.2 IU mg-1. The resultant enzyme is 40% more stable than commercially available Escherichia coli's ASNase (EcASNaseII) when incubated with human serum. In addition, ScASNaseII presents 50% lower cross-reaction with anti-ASNase antibody produced against EcASNaseII when compared with ASNase from Dickeya chrysanthemi.

A Saccharomyces eubayanus haploid resource for research studies.

Since its identification, Saccharomyces eubayanus has been recognized as the missing parent of the lager hybrid, S. pastorianus. This wild yeast has never been isolated from fermentation environments, thus representing an interesting candidate for evolutionary, ecological and genetic studies. However, it is imperative to develop additional molecular genetics tools to ease manipulation and thus facilitate future studies. With this in mind, we generated a collection of stable haploid strains representative of three main lineages described in S. eubayanus (PB-1, PB-2 and PB-3), by deleting the HO gene using CRISPR-Cas9 and tetrad micromanipulation. Phenotypic characterization under different conditions demonstrated that the haploid derivates were extremely similar to their parental strains. Genomic analysis in three strains highlighted a likely low frequency of off-targets, and sequencing of a single tetrad evidenced no structural variants in any of the haploid spores. Finally, we demonstrate the utilization of the haploid set by challenging the strains under mass-mating conditions. In this way, we found that S. eubayanus under liquid conditions has a preference to remain in a haploid state, unlike S. cerevisiae that mates rapidly. This haploid resource is a novel set of strains for future yeast molecular genetics studies.

Inhibitory effect of different probiotic formulations on Salmonella Heidelberg, avian pathogenic Escherichia coli and Campylobacter jejuni

A wide variety of microorganism species have been used as probiotics to improve the intestinal microbial balance, control and prevent the colonization of pathogenic bacteria and promote growth in animal production. Based on the various beneficial factors of probiotic agents, this study aimed to evaluate the effectiveness of three different formulations of probiotics strains through the analysis of antagonistic capacity against common pathogens from chickens gastrointestinal tract. Three formulations composed by Bacillus, Lactobacillus, Saccharomyces, Enterococcus, Bifidobacterium and Pediococcus were tested in vitro by the method of probiotic culture spot in plates seeded with inocula of Escherichia coli, Salmonella Heidelberg and Campylobacter jejuni. The inhibition halos were measured and classified according to the degree of pathogenic bacteria inhibition. The formulation containing an association among Bacillus, Lactobacillus and Saccharomyces presented better results when compared to the other formulations.

Sour beer production in India using a coculture of Saccharomyces pastorianus and Lactobacillus plantarum: optimization, microbiological, and biochemical profiling.

The study's objective was to develop a co-fermentation process with appropriate fermentation parameters to produce a sour beer (similar to a Belgium sour beer) with an ethanol content of 6-8% (v/v) using a coculture of Saccharomyces pastorianus and Lactobacillus plantarum. Statistical optimization was conducted to determine fermentation conditions to produce a sour beer with ~ 3 mg/mL of lactic acid, similar to the traditional sour beer levels. Studies were conducted on the microbial dynamics and volatile compounds produced during this fermentation and aging process. GC-MS studies revealed the generation of novel bioactive compounds as well as the depletion of some volatile compounds during co-fermentation. The study detailed a 5-day co-fermentation process of S. pastorianus and L. plantarum and a 21-day aging process to prepare a sour beer with biochemical properties along the lines of traditional lambic beers. The interrelationship between the two microorganisms and the biochemical changes in the sour beer fermentation process was elucidated and the sensorial attributes have been described.

Effect of non-wine Saccharomyces yeasts and bottle aging on the release and generation of aromas in semi-synthetic Tempranillo wines.

Interest in the use of non-conventional yeasts in wine fermentation has been increased in the last years in the wine sector. The main objective of this manuscript was to explore the aromatic diversity produced by wild and non-wine strains of S. cerevisiae, S. eubayanus, S. kudriavzevii, and S. uvarum species in young and bottle-aged Tempranillo wines as well as evaluate their fermentation capacity and the yield on ethanol, glycerol, and organic acids, that can contribute to diminishing the effects of climate change on wines. S. uvarum strain U1 showed the highest ability to release or de novo produce monoterpenes, such as geraniol and citronellol, whose values were 1.5 and 3.5-fold higher than those of the wine S. cerevisiae strain. We found that compared to the normal values for red wines, ß-phenylethyl acetate was highly synthesized by U1 and E1 strains, achieving 1 mg/L. Additionally, after aging, wines of S. eubayanus strains contained the highest levels of this acetate. Malic acid was highly degraded by S. kudriavzevii yeasts, resulting in the highest yields of lactic acid (>5-fold) and ethyl lactate (>2.8-fold) in their wines. In aged wines, we observed that the modulating effects of yeast strain were very high in ß-ionone. S. uvarum strains U1 and BMV58 produced an important aging attribute, ethyl isobutyrate, which was highly enhanced during the aging. Also, the agave S. cerevisiae strain develops an essential aroma after aging, reaching the highest ethyl leucate contents. According to the results obtained, the use of wild non-wine strains of S. cerevisiae and strains of the cryotolerant species S. eubayanus, S. kudriavzevii, and S. uvarum in Tempranillo wine fermentation increase the aroma complexity. In addition, wines from S. kudriavzevii strains had twice additional glycerol, those from S. uvarum 4-fold more succinic acid, while wines from wild strains yielded 1% v/v less ethanol which may solve wine problems associated with climate change.

Rapid selection response to ethanol in Saccharomyces eubayanus emulates the domestication process under brewing conditions.

Although the typical genomic and phenotypic changes that characterize the evolution of organisms under the human domestication syndrome represent textbook examples of rapid evolution, the molecular processes that underpin such changes are still poorly understood. Domesticated yeasts for brewing, where short generation times and large phenotypic and genomic plasticity were attained in a few generations under selection, are prime examples. To experimentally emulate the lager yeast domestication process, we created a genetically complex (panmictic) artificial population of multiple Saccharomyces eubayanus genotypes, one of the parents of lager yeast. Then, we imposed a constant selection regime under a high ethanol concentration in 10 replicated populations during 260 generations (6 months) and compared them with propagated controls exposed solely to glucose. Propagated populations exhibited a selection differential of 60% in growth rate in ethanol, mostly explained by the proliferation of a single lineage (CL248.1) that competitively displaced all other clones. Interestingly, the outcome does not require the entire time-course of adaptation, as four lineages monopolized the culture at generation 120. Sequencing demonstrated that de novo genetic variants were produced in all propagated lines, including SNPs, aneuploidies, INDELs and translocations. In addition, the different propagated populations showed correlated responses resembling the domestication syndrome: genomic rearrangements, faster fermentation rates, lower production of phenolic off-flavours and lower volatile compound complexity. Expression profiling in beer wort revealed altered expression levels of genes related to methionine metabolism, flocculation, stress tolerance and diauxic shift, likely contributing to higher ethanol and fermentation stress tolerance in the evolved populations. Our study shows that experimental evolution can rebuild the brewing domestication process in 'fast motion' in wild yeast, and also provides a powerful tool for studying the genetics of the adaptation process in complex populations.

Evaluation of cryotolerant yeasts for the elaboration of a fermented pear beverage in Patagonia: Physicochemical and sensory attributes.

The production of pome fruits as pears and apples, as well as their derived industries, is of great economic importance in North Patagonia. The elaboration of fermented beverages as cider or perry has evidenced a substantial diversification during the last years, with the evaluation of different fruit varieties, yeast starters and technological changes. In this work, two cryotolerant yeasts belonging to the species Saccharomyces uvarum were evaluated at laboratory and pilot scale in sterile and no-sterile pear must. One of the strains was originally isolated from apple chicha (strain NPCC1314) and the other from apple cider (strain NPCC1420) in Patagonia. Both physicochemical and sensory features of the fermented products were evaluated. Both strains were able to successfully complete the fermentations, although strain NPCC1420 showed the better kinetic properties including a faster sugar consumption than the strain NPCC1314. Both strains showed excellent implantation capacity, but the fermented products showed different chemical profiles. The perry fermented with the strain NPCC1314 was characterized by better sensory attributes as assessed by trained panelists and a greater acceptance for untrained public than the same fermented with the strain NPCC1420. The two strains were able to consume sorbitol, both in pear must and in agar-plates supplemented with sorbitol as the sole carbon source. This ability is described for the first time in S. uvarum, at least for the two strains evaluated in this work.

Effect of Saccharomyces and non-Saccharomyces native yeasts on beer aroma compounds.

Recently, the increase in microbreweries and the consequent production of craft beers have reached exponential growth. The interest in non-conventional yeasts for innovation and a unique selling feature in beer fermentation is increasing. This work studied the autochthonous Saccharomyces and non-Saccharomyces yeasts, isolated from various food sources, with the ability to modify and improve the fermentative and aromatic profiles during alcoholic fermentation. The ability to ferment maltose and produce desirable aroma compounds were considered as the key characters for the screening selection. A synthetic beer wort was developed for this purpose, to simulate beer wort composition. A total of forty-seven yeast strains belonging to different genera were analysed according to their fermentation profile, volatile compounds production and sensory analysis. Three native strains of Saccharomyces cerevisiae, Zygoascus meyerae and Pichia anomala were selected to evaluate their aromatic profile in single and mixed fermentations. The strains produced 4-vinylguaiacol, ß-phenylethyl alcohol, and isoamyl alcohol at levels significantly above the sensory threshold, making them interesting for wheat and blond craft beer styles. The native Hanseniaspora vineae was also included in a co-fermentation treatment, resulting in a promising yeast to produce fruity beers.

Research of antifungal effects on some essential oils with tube dilution / Pesquisa de efeitos antifúngicos em alguns óleos essenciais com diluição em tubo

The aim of study to investigate inhibition effects of cinnamic acid (volatile compound of cinnamon), limonene (essential oil of lemon) and eugenol (essential oil of clove, cinnamon) on Metschnikowia fructicola, Candida oleophila, Schisosaccharomyces pombe, Saccharomyces uvarum and Kloeckera apiculata. In this study, tube dilution method was used. Among eugenol, limonene and cinnamic acid, it was found that cinnamic acid has the most inhibition effect with low concentrations (%2.8, %3.84, %4.36, %5,4) at tube dilution methods. Also it was found that test yeast have different resistance against test materials.(AU)

O objetivo do estudo foi investigar os efeitos de inibição do ácido cinâmico (composto volátil da canela), limoneno (óleo essencial de limão) e eugenol (óleo essencial de cravo, canela) sobre Metschnikowia fructicola, Candida oleophila, Schisosaccharomyces pombe, Saccharomyces uvarum e Kloeckera apiculata. Neste estudo, foi utilizado o método de diluição em tubo. Entre eugenol, limoneno e ácido cinâmico, verificou-se que o ácido cinâmico tem o maior efeito de inibição com baixas concentrações (% 2,8,% 3,84,% 4,36,% 5,4) nos métodos de diluição em tubo. Também foi descoberto que a levedura testada tem diferentes resistências contra os materiais de teste.(AU)

Use of Saccharomyces cerevisiae-based products and effects on rumen environment and performance of sheep subjected to dietary changes / Utilização de produtos à base de Saccharomyces cerevisiae e seus efeitos sobre o ambiente ruminal e desempenho de ovinos submetidos a mudanças de dieta

Natural additives in ruminant diet optimizes animal performance by controlling or modifying the rumen fermentation pattern. The aim of this study was to evaluate the effects of using Saccharomyces cerevisiae products during abrupt changes in dietary starch concentration on ruminal pH maintenance, performance and metabolism of confined sheep. Twenty adult ewes, crossbred Texel x Corriedale(44.7±6.97 Kg BW) were divided into three groups: Control (n=6), Cultron X (yeast culture; n=7) and Cultron Pro (hydrolyzed yeast; n=7), administered at a dose of 5 g/sheep/day (10x1010CFU/g DM). The experiment lasted 20 days divided into four periods, alternating between 40 and 60% concentrate. Blood and ruminal fluid were collected at the end of each period, and daily dry matter intake was measured to determine the average daily gain (ADG). Blood tests included blood count and biochemical parameters such as glucose, urea, total plasma proteins (TPP), liver enzymes, acute phase proteins and minerals. There was an upward trend in ADG in group Cultron X compared to control in period 3. Although, ruminal pH did not differ between groups, protozoan motility was higher in animals supplemented with hydrolyzed yeast. The Cultron Pro group showed an increase in TPP in periods 3 and 4, and AST in periods 2 and 3 compared to the control group. There was no difference in other metabolites. Yeast supplementation is a potential tool in times of feeding challenge, in which yeast culture is the most effective for improving performance of confined sheep being in improving performance of confined sheep.(AU)

A utilização de aditivos naturais na dieta de ruminantes é uma prática que otimiza o desempenho, controlando ou modificando o padrão de fermentação ruminal. O objetivo do estudo buscou avaliar os efeitos da utilização de produtos à base de Saccharomyces cerevisiae durante mudanças na concentração de amido da dieta sobre a manutenção do pH ruminal, desempenho e metabolismo de ovinos confinados. Dessa forma, 20 fêmeas ovinas adultas mestiças das raças Texel e Corriedale (44,7±6,97 PV) foram divididas em três grupos: Controle (n=6), Cultron X (cultura de levedura; n=7) e Cultron Pro (levedura hidrolisada; n=7), administrados na dose de 5 g/ovelha/dia (10x1010ufc/g de MS). O experimento teve duração de 20 dias divididos em quatro períodos, alternando entre 40 e 60% de concentrado. Foram realizadas coletas de sangue e líquido ruminal ao final de cada período e mensuração da ingestão de matéria seca diariamente, a fim de determinar o ganho médio diário (GMD). As análises sanguíneas incluíram hemograma e parâmetros bioquímicos como glicose, ureia, proteínas plasmáticas totais (PPT), enzimas hepáticas, proteínas de fase aguda e minerais. Houve tendência de aumento no GMD no grupo Cultron X em relação ao controle no período 3. Apesar do pH ruminal não diferir entre grupos, a motilidade de protozoários foi melhor nos animais suplementados com levedura hidrolisada. O grupo Cultron Pro apresentou aumento de PPT nos períodos 3 e 4 e aumento nos níveis da enzima hepática AST nos períodos 2 e 3 em comparação ao grupo controle. Não houve diferença nos demais metabólitos. A suplementação com levedura apresentou-se como potencial ferramenta em momentos de desafio alimentar, sendo a cultura de levedura mais eficaz na melhora da performance de ovinos confinados.(AU)

Efeitos do probiótico Saccharomyces ceveriseae no tratamento da periodontite experimental em ratos submetidos à quimioterapia / Effects of the probiotic Saccharomyces Ceveriseae in the treatment of experimental periodontitis in rats undergoing chemotherapy

O presente estudo teve a finalidade de avaliar os efeitos do uso do probiótico (PRO) Saccharomyces ceveriseae coadjuvante à raspagem e alisamento radicular (RAR) no tratamento da periodontite experimental (PE) induzida em animais imunossuprimidos com o quimioterápico 5-Fluorouracil (5FU). Cento e oito animais foram submetidos à indução da PE por meio da instalação de um fio de algodão ao redor do primeiro molar inferior esquerdo ou direito. A imunossupressão foi obtida pela injeção intraperitoneal do 5FU aplicada em dois momentos: no ato da instalação da ligadura e após 48 horas. Sete dias após instalação a ligadura foi removida e os animais separados aleatoriamente em seis grupos experimentais que receberam os seguintes tratamentos: PE (n=18) - animais que não sofreram nenhum tratamento sistêmico ou local; SS (n=18) - animais que não sofreram tratamento local, apenas tratamento sistêmico com solução salina (SS); 5FU (n=18) - animais tratados sistemicamente com 5FU que não sofreram nenhum tratamento local; 5FU-PRO (n=18) - animais tratados sistemicamente com 5FU que receberam irrigação local com 0,6 ml de PRO; 5FURAR (n=18) - animais tratados sistemicamente com 5FU que receberam RAR e; 5FURAR-PRO (n=18) - animais tratados sistemicamente com 5FU, que receberam tratamento com RAR e irrigação local com 0,6 ml de PRO. Seis animais de cada período foram submetidos à eutanásia aos 7, 15 e 30 dias após os tratamentos. Os espécimes contendo a área interessada foram obtidos e processados para a análise histopatológica, histométrica da perda óssea alveolar (POA), porcentagem de tecido ósseo (PTO), porcentagem de osso vital (POV) e porcentagem de osso não vital (PONV), e análise imunoistoquímica (TRAP, RANKL e OPG). Os dados obtidos foram submetidos à análise estatística (α=5%). Os resultados histológicos demonstraram que nos espécimes tratados com 5FU houve agravamento da perda óssea na região de furca comparado com os animais sistemicamente saudáveis (grupos PE e SS); que o tratamento com PRO como monoterapia (grupo 5FU-PRO) promoveu redução do processo inflamatório, denotando estabilização da doença periodontal; que o tratamento com RAR (grupo 5FU-RAR) reduziu o volume do infiltrado inflamatório mostrando sinais de reparação e a associação PRO-RAR (grupo 5FU-RAR-PRO) evidenciou nítida reestruturação tecidual aos 15 dias e menor área de reabsorção óssea aos 30 dias. Observou-se na análise histométrica que a POA foi maior no grupo 5FU-PRO em relação ao grupo SS aos 30 dias (p<0,05); a PTO foi maior no grupo 5FU-RAR-PRO aos 30 dias em comparação aos grupos EP, SS, 5FU e 5FU-PRO (p<0,05) e a POV foi maior no grupo 5FU-RAR-PRO em comparação ao grupo 5FU aos 7 dias, e maior aos 30 dias em comparação a todos os grupos experimentais (p<0,05). Não houve diferença entre os grupos e períodos na imunomarcação das células TRAP positivas. A RANKL mostrou alto padrão de imunomarcação nos grupos PE, SS e 5FU em todos períodos, e aos 7 dias no grupo 5FU-PRO, 5FU-RAR e 5FURAR-PRO, moderado padrão de imunomarcação nos grupos 5FU, 5FU-PRO, 5FURAR, e 5FU-RAR-PRO, mantendo-se baixo padrão neste último grupo, no período de 30 dias. Houve baixo padrão de imunomarcação de OPG em todos os grupos e períodos, exceto no grupo 5FU-RAR-PRO que mostrou moderado padrão. Diante dos resultados obtidos pode ser concluído que o uso do quimioterápico 5FU agravou a PE; que o uso do Saccharomyces ceveriseae associado à RAR, minimizou os efeitos do quimioterápico nos tecidos periodontais reduzindo o processo inflamatório, e reduzindo a porcentagem de destruição de tecido ósseo na área de furca em animais imunossuprimidos(AU)

The present study aimed to evaluate the effects of using probiotic (PRO) Saccharomyces ceveriseae as an adjuvant of scaling and root planing (SRP) in the treatment of experimental periodontitis (EP) induced in immunosuppressed animals with chemotherapeutic 5-Fluorouracil (5FU). One hundred and eight animals were submitted to EP induction by installing a cotton thread around the lower left or right first molar. Immunosuppression was obtained by intraperitoneal injection of 5FU applied in two moments: at the time of installing the ligature and after 48 hours. Seven days after installation, the ligature was removed and the animals were randomly separated into six experimental groups that received the following treatments: EP (n = 18) - animals that did not receive any systemic or local treatment; SS (n = 18) - animals that did not receive local treatment, only systemic treatment with saline solution (SS); 5FU (n = 18) - animals treated systemically with 5FU that did not receive any local treatment; 5FUPRO (n = 18) - animals treated systemically with 5FU that received local irrigation with 0.6 ml of PRO; 5FU-SRP (n = 18) - animals treated systemically with 5FU that received RAR and; 5FU-SRP-PRO (n = 18) - animals treated systemically with 5FU, which received treatment with SRP and local irrigation with 0.6 ml of PRO. Six animals from each period were euthanized at 7, 15 and 30 days after treatments. The specimens containing the interested area were obtained and processed for histopathological, histometric analysis of alveolar bone loss (ABL), percentage of bone tissue (PBT), percentage of vital bone (PVB) and percentage of non-vital bone (PNVB), and immunohistochemical analysis (TRAP, RANKL and OPG). The data obtained were subjected to statistical analysis (α = 5%). The histological results showed that in the specimens treated with 5FU there was worsening of bone loss in the furcation region compared to healthy animals (EP and SS groups); that treatment with PRO as monotherapy (group 5FU-PRO) reduced the inflammatory process, denoting stabilization of periodontal disease; that treatment with SRP (5FU-SRP group) reduced the volume of the inflammatory infiltrate showing signs of repair and the PRO-SRP association (5FU-SRP-PRO group) showed a clear tissue restructuring at 15 days and a smaller area of bone resorption at 30 days. It was observed in the histometric analysis that ABL was higher in the 5FU-PRO group compared to SS group at 30 days (p <0.05); PTO was higher in 5FU-SRP-PRO group at 30 days compared to EP, SS, 5FU and 5FU-PRO groups (p <0.05) and PVB was higher in the 5FU-SRP-PRO group compared to 5FU group at 7 days, and greater at 30 days compared to all experimental groups (p <0.05). There was no difference between groups and periods in the immunostaining of TRAP positive cells. RANKL showed a high immunostaining pattern in the EP, SS and 5FU groups in all periods, and at 7 days in 5FU-PRO, 5FU-SRP and 5FU-SRP-PRO groups, a moderate immunostaining pattern in the 5FU, 5FU-PRO, 5FU-SRP and 5FU-SRP-PRO groups, maintaining a low standard in 5FU-SRP-PRO group, in the period of 30 days. There was a low pattern of OPG immunostaining in all groups and periods, except in 5FU-SRP-PRO group, which showed a moderate pattern. In view of the results obtained, it can be concluded that the use of 5FU chemotherapy has worsened the evolution of EP; that the use of Saccharomyces ceveriseae associated with RAR, minimized the effects of chemotherapy on periodontal tissues, reducing the inflammatory process, and reducing the percentage of bone tissue destruction in the furcation area in immunosuppressed animals(AU)

Efectos de la adición de probiótico Saccharomyces cerevisiae sobre histomorfología intestinal en pollos de engorde / Effects of addition of probiotic Saccharomyces cerevisiae on intestinal histomorphology in broilers

RESUMEN Este trabajo describe los efectos del probiótico Saccharomyces cerevisiae sobre el área, número de criptas de Lieberkühn en duodeno y yeyuno, y producción de moco en ambas secciones intestinales de pollos de engorde. Fueron empleados los tejidos de un total de 27 individuos clasificados en un grupo control GC (n=12) y un grupo suplementado con probióticos GP (n=15). Los resultados revelaron que los grupos suplementados con el S. cerevisiae presentaron una mayor amplitud del área de las criptas en duodeno (p= 0,0119) y yeyuno (p= 0,0355), menor número de criptas por milímetro en duodeno (p= 0,0420) y mayor producción de moco en duodeno respecto al grupo control (p= 0,0185), mientras que en yeyuno no se observaron diferencias significativas. Se concluyó que el uso de Saccharomyces cerevisiae aumentó el tamaño del área de las criptas en ambas secciones intestinales y aumentó la producción de moco en duodeno; lo cual, al aumentar la superficie de absorción intestinal, seguramente podría resultar en mejoras de los parámetros productivos.

ABSTRACT This work describes the effects of the probiotic Saccharomyces cerevisiae on the area, number of Lieberkühn crypts in the duodenum and jejunum, and mucus production in both intestinal sections. Tissues from a total of 27 individuals were used, classified in control group - GC (n = 12) and group supplemented with probiotics - GP (n = 15). The results revealed that the group supplemented with S. cerevisiae presented a greater area of the crypts in the duodenum (p = 0.0119) and jejunum (p = 0.0355), less number of crypts per millimeter in the duodenum (p = 0.0420) and higher mucus production in the duodenum compared to the control group (p = 0.0185), while in the jejunum no significant differences were observed. It was concluded that the use of Saccharomyces cerevisiae increased the size of the crypt area in both intestinal sections and increased mucus production in the duodenum; which by increasing the intestinal absorption surface could surely result in improvements in the productive parameters.

An Out-of-Patagonia migration explains the worldwide diversity and distribution of Saccharomyces eubayanus lineages.

Population-level sampling and whole-genome sequences of different individuals allow one to identify signatures of hybridization, gene flow and potential molecular mechanisms of environmental responses. Here, we report the isolation of 160 Saccharomyces eubayanus strains, the cryotolerant ancestor of lager yeast, from ten sampling sites in Patagonia along 2,000 km of Nothofagus forests. Frequency of S. eubayanus isolates was higher towards southern and colder regions, demonstrating the cryotolerant nature of the species. We sequenced the genome of 82 strains and, together with 23 available genomes, performed a comprehensive phylogenetic analysis. Our results revealed the presence of five different lineages together with dozens of admixed strains. Various analytical methods reveal evidence of gene flow and historical admixture between lineages from Patagonia and Holarctic regions, suggesting the co-occurrence of these ancestral populations. Analysis of the genetic contribution to the admixed genomes revealed a Patagonian genetic origin of the admixed strains, even for those located in the North Hemisphere. Overall, the Patagonian lineages, particularly the southern populations, showed a greater global genetic diversity compared to Holarctic and Chinese lineages, in agreement with a higher abundance in Patagonia. Thus, our results are consistent with a likely colonization of the species from peripheral glacial refugia from South Patagonia. Furthermore, fermentative capacity and maltose consumption resulted negatively correlated with latitude, indicating better fermentative performance in northern populations. Our genome analysis, together with previous reports in the sister species S. uvarum suggests that a S. eubayanus ancestor was adapted to the harsh environmental conditions of Patagonia, a region that provides the ecological conditions for the diversification of these ancestral lineages.