Background: The control of energy metabolism is fundamental for cell growth and function, and anomalies are implicated in complex diseases and ageing. It is important for cells to carefully tune metabolic pathways to optimize their function in response to different nutrient or physiological conditions. Metabolism in yeast cells can be easily manipulated by supplying different carbon sources: on glucose yeast rapidly proliferates by fermentation, analogous to tumour cells growing by aerobic glycolysis, whereas on non-fermentable carbon sources metabolism shifts towards respiration. Results: We screened deletion libraries of fission yeast to identify over 200 genes required for respiratory growth. The growth medium and auxotrophic mutants strongly influenced respiratory metabolism. Most genes uncovered in the mutant screens have not been implicated in respiration in budding yeast. We applied gene expression profiling approaches to compare steady-state fermentative and respiratory growth and to analyse the dynamic adaptation to respiratory growth. The transcript levels of most genes functioning in key energy metabolism pathways were coherently tuned, reflecting anticipated differences in metabolic flows between fermenting and respiring cells. We show that the acetyl-CoA synthase, rather than the citrate lyase, is essential for acetyl-CoA synthesis in fission yeast. We also investigated the transcriptional response to mitochondrial damage by genetic or chemical perturbations, defining a retrograde response that involves the concerted regulation of distinct groups of nuclear genes that may avert harm from mitochondrial malfunction. Conclusions: These systematic and targeted analyses provide a rich framework of the genetic and regulatory basis of fundamental metabolic states to guide future studies on energy metabolism in fission yeast and beyond. Our study pinpoints weaknesses of commonly used auxotroph mutants for investigating energy metabolism. As a model for cellular energy regulation, fission yeast provides an attractive and complementary system to budding yeast.

To advance probiotic research, a comprehensive understanding of bacterial interactions with human physiology at the molecular and cellular levels is fundamental. Lacticaseibacillus rhamnosus LGG® is a bacterial strain that has long been recognized for its beneficial effects on human health. Probiotic effector molecules derived from LGG®, including secreted proteins, surface-anchored proteins, polysaccharides, and lipoteichoic acids, that interact with host physiological processes have been identified. In vitro and animal studies have revealed that specific LGG® effector molecules stimulate epithelial cell survival, preserve intestinal barrier integrity, reduce oxidative stress, mitigate excessive mucosal inflammation, enhance IgA secretion, and provide long-term protection through epigenetic imprinting. Pili on the cell surface of LGG® promote adhesion to the intestinal mucosa and ensure close contact to host cells. Extracellular vesicles produced by LGG® recapitulate many of these effects through their cargo of effector molecules. Collectively, the effector molecules of LGG® exert a significant influence on both the gut mucosa and immune system, that promotes intestinal homeostasis and immune tolerance.

The global craft beer market is witnessing substantial growth, with a particular spotlight on sour beers. These unique brews are created through lactic fermentation initiated by lactic acid bacteria (LAB). However, the presence of hops, renowned for their antimicrobial properties, can pose a limitation within the realm of sour beer production. This research aims to explore and assess the potential of sour beer as a probiotic beverage. It involves a comparative analysis of the viability of Lacticaseibacillus paracasei subsp. paracasei F19 (F19) and 431 (L431) in a sour beer with an International Bitterness Unit (IBU) value of 26.65, alongside examining the ex-pression of genes such as horA, horB, horC, hitA, bsrA, and recA, which are associated with hop resistance. The study encompasses the development of two formulations of sour beers characterized by both high hop content and a robust probiotic count (8.44 – 8.77 log CFU/mL). The findings suggest that both probiotic strains of F19 and L431, are well-suited to produce sour beers with elevated hop levels, demonstrating excellent stability. Notably, the expression of genes responsible for hops resistance exhibited distinct modulation patterns among the two strains. It appears that a higher concentration of bsrA, as observed in strain L431, may be more effective in mitigating the effects of hop-related stress, potentially surpassing the combined expression of horA and bsrA. Nonetheless, further research is essential to validate this observation in gene expression. F19 and L431 emerge as promising candidates for the development of sour beers with increased hop intensity.

Diarrhoea is a serious cause of mortality worldwide that can lead to dehydration, gut barrier function impairment, nutrient malabsorption, and alterations of the gut microbiota (dysbiosis). Current solutions for its management, such as oral rehydration salts (ORS), inhibitors of the gut motility, antibiotics, and living probiotics only partially counteract the mechanisms of the disease and do not provide a full coverage of the problem. Potential risks of the use of living probiotic strains, particularly in immunocompromised patients, can be eliminated with the use of tyndallized (heat-killed) postbiotic bacteria and yeast. ABB C22® is a postbiotic combination of three tyndallized yeasts, namely Saccharomyces boulardii, Saccharomyces cerevisiae, and Kluyveromyces marxianus. To assess the action of the postbiotic combination on diarrhoea, immune and gut epithelial cell signaling assays, gut barrier formation assay and rotavirus gene expression assay were performed. ABB C22® showed a strong anti-inflammatory effect, an induction of the build-up of the gut epithelium, and a degree of protection against rotavirus infection. These experimental studies support the use of the postbiotic ABB C22® as a solution for the management of diarrhoea and gastrointestinal conditions, alone or in combination with existing but incomplete treatments.

Diarrhoea is a serious cause of mortality worldwide that can lead to dehydration, gut barrier function impairment, nutrient malabsorption, and alterations of the gut microbiota (dysbiosis). Current solutions for its management, such as oral rehydration salts (ORS), inhibitors of the gut motility, antibiotics, and living probiotics only partially counteract the mechanisms of the disease and do not provide a full coverage of the problem. Potential risks of the use of living probiotic strains, particularly in immunocompromised patients, can be eliminated with the use of tyndallized (heat-killed) postbiotic bacteria and yeast. ABB C22® is a postbiotic combination of three tyndallized yeasts, namely Saccharomyces boulardii, Saccharomyces cerevisiae, and Kluyveromyces marxianus. To assess the action of the postbiotic combination on diarrhoea, immune and gut epithelial cell signaling assays, gut barrier formation assay and rotavirus gene expression assay were performed. ABB C22® showed a strong anti-inflammatory effect, an induction of the build-up of the gut epithelium, and a degree of protection against rotavirus infection. These experimental studies support the use of the postbiotic ABB C22® as a solution for the management of diarrhoea and gastrointestinal conditions, alone or in combination with existing but incomplete treatments.

Saccharomyces cerevisiae are a phenotypically diverse species that adapt to a wide variety of environments by exploiting standing genetic diversity and selecting for advantageous mutations. Glyphosate and copper-based herbicides/ fungicides affect non-target organisms, these incidental exposures can impact microbial populations. In this study, glyphosate resistance was found in the historical collection of yeast which was collected over the last century, but only in yeast isolated after the introduction of glyphosate. The highest glyphosate-resistant yeasts were isolated from agricultural sites. However, herbicide application at these sites was not recorded. In an effort to assess glyphosate resistance and impact on non-target microorganisms, yeast were harvested from 15 areas with known herbicidal histories, including an organic farm, conventional farm, remediated coal mine, suburban locations, state park, and a national forest. Yeast representing 23 genera were isolated from 237 samples of plant, soil, spontaneous fermentation, nut, flower, fruit, feces, and tree material samples. Saccharomyces, Candida, Metschnikowia, Klyveromyces, Hanseniaspora, and Pichia were other genera commonly found across our sampled environments. Managed areas had less species diversity and at the brewery, only Saccharomyces and Pichia were isolated. A conventional farm growing RoundUp Ready™ corn had the lowest phylogenetic diversity and the highest glyphosate resistance. The mine was sprayed with multiple herbicides including a commercial formulation of glyphosate; however, the yeast did not have elevated glyphosate resistance. In contrast to the conventional farm, the mine was exposed to glyphosate only one year prior to sample isolation. Glyphosate resistance is an example of the anthropogenic selection of nontarget organisms.

Yeast, crucial in beer production, holds promise as a valuable by-product resource, known as brewer's spent yeast (BSY), with potentially beneficial physiological effects. This study aimed to compare the composition and soluble polysaccharide content of Brewer's spent yeast with those of pure cultured yeast strains, namely Saccharomyces cerevisiae (SC) and S. boulardii (SB), to facilitate the utilization of BSY as an alternative source of functional polysaccharides. BSY exhibited significantly higher carbohydrate content and lower crude protein content than SC and SB cells. The residues recovered through autolysis were 53.11%, 43.83%, and 44.99% for BSY, SC, and SB, respectively. Notably, the polysaccharide content of BSY residue (641.90 μg/mg) was higher than that of SC (553.52 μg/mg) and SB (591.56 μg/mg). The yields of alkali-extracted water-soluble polysaccharides were 40%, 30%, and 35% for BSY, SC, and SB, respectively, with BSY comprising a higher proportion of water-soluble saccharides than SC and SB, including 49.3% mannan and 20.1% β-glucan. Furthermore, BSY demonstrated antioxidant activities, including superoxide dismutase (SOD), ABTS, and DPPH scavenging potential, suggesting its ability to mitigate oxidative stress. BSY also exhibited a significantly higher total phenolic compound content, indicating its potential as an effective functional food material.

Probiotic feed additives have attracted considerable research interest in recent years because the effectiveness of probiotics can differ across microbial strains and the supplemented macroorganisms. The present study was conducted on 16 lambs divided into two groups (C - control and E - experimental). The examined lambs were aged 11 days at the beginning of the experiment and 40 days at the end of the experiment. The diet of group E lambs was supplemented with a multi-strain probiotic formulation (Lactobacillus plantarum AMT14, Lactobacillus plantarum AMT4, Lactobacillus rhamnosus AMT15, and Bifidobacterium animalis AMT30), whereas group C lambs did not receive the probiotic additive. At the beginning of the experiment (day 0) and on experimental days 15 and 30, blood was sampled from the jugular vein to determine and compare: phagocytic activity (PHAGOTEST) and oxidative metabolism (PHAGOBURST) of peripheral blood granulocytes and monocytes by flow cytometry. An analysis of the phagocytic activity of granulocytes and monocytes revealed significantly higher levels of phagocytic activity (expressed as the percentage of phagocytic cells and mean fluorescence intensity) in lambs administered the multi-strain probiotic formulation than in the control group. The probiotic feed additive also exerted a positive effect on the oxidative metabolism of both granulocytes and monocytes (expressed as the percentage of oxidative metabolism and mean fluorescence intensity) after stimulation with Escherichia coli bacteria and with PMA (4-phorbol-12-β-myristate-13-acetate).

: Microbiology has long been a keystone in fermentation and the utilization of yeast biology rein-forces molecular biotechnology as the pioneering frontier in brewing science. Consequently, modern understanding of the brewer’s yeast has faced significant refinement over the last few decades. This publication presents a condensed summation of Saccharomyces species dynamics with an emphasis on the relationship between traditional ale yeast, Saccharomyces cerevisiae, and the interspecific hybrids used in lager beer production, S. pastorianus. Introgression from other Sac-charomyces species is also touched on. The unique history of Saccharomyces cerevisiae and Saccharo-myces hybrids are exemplified by recent genomic sequencing studies aimed at categorizing brewing strains through phylogeny and redefining Saccharomyces species boundaries. Phylogenetic investigations highlight the genomic diversity of Saccharomyces cerevisiae ale strains long known to brewers by their fermentation characteristics and phenotypes. Discoveries of genomic contribu-tions from interspecific Saccharomyces species into the genome of S. cerevisiae strains is ever more apparent with increased investigations on the hybrid nature of modern industrial and historical fermentation yeast.

Biocontainment techniques for genetically modified yeasts (GMYs) are pivotal due to the importance of these organisms for biotechnological processes and also due to the design of new yeast strains by using synthetic biology tools and technologies. Due to the large genetic modifications that many yeast strains display, it is highly desirable to avoid the leakage of GMY cells into natural environments and, consequently, the spread of synthetic genes and circuits by horizontal or vertical gene transfer mechanisms within the microorganisms. Moreover, it is also desirable to avoid that patented yeast gene technologies spread outside the production facility. In this review, it was evaluated the different biocontainment technologies currently available for GMYs. Interestingly, uniplex-type biocontainment approaches (UTBAs), which relies on nutrient auxotrophies induced by gene mutation or deletion, or the expression of simple kill switches apparatus, are still the major biocontainment approaches still in use with GMY. While bacteria like Escherichia coli account for advanced biocontainment technologies based on synthetic biology and multiplex-type biocontainment approaches (MTBAs), GMYs are distant from this scenario due to many reasons. Thus, a comparison of different UTBAs and MTBAs applied for GMY and genetically engineered microorganisms (GEMs) was made, indicating the major advances of biocontainment techniques for GMYs.

A bacterium known as Akkermansia muciniphila (AkkermansiaA. muciniphila) was detected in human feces in 2004. It is defined as a gram-negative, anaerobic, and mucin-degrading microbe belonging to the Verrucomicrobia species. Although the majority of A. muciniphila is found in the human intestinal tract playing a role in the gut microbiome, there are also smaller levels detected in breast milk and oral cavities as well as in the pancreas and appendix.Evidence suggests that having A. muciniphila present in the gut microbiome has positive effects on human health. As a result, A. muciniphila has been shown to have health benefits when taken as a dietary supplementation where it is considered a promising next-generation probiotic. However, the large-scale production of A. muciniphila remains a challenge due to its need for a stable environment. While culturing A. muciniphila in a mucin-based medium is one option, other strategies that use plate-based mediums have also been explored. Currently, both live and pasteurized forms of A. muciniphila are available on the market and shown to have promising outcomes in animal models. Additionally, researchers are investigating various prebiotics to increase the abundance of A. muciniphila in the human gut. This article presents a detailed summary of the phenotypic characteristics of A. muciniphila and its associations with various human health condition illnesses. Finally, the paper explores A. muciniphila's potential as an effective next-generation probiotic.

Yeast extract is widely used in different food industries as a flavoring agent or vitamin supplement. In this study, a process was optimized for the production of yeast extract from Baker’s yeast (Saccharomyces cerevisiae). A glass vessel stirred fermenter was used for the cultivation of yeast biomass. The effect of various physical and chemical factors was evaluated on the production of yeast cells and optimum conditions for the production of maximum yeast biomass were determined. The optimum growth was obtained at 30ºC with pH 4.5 using molasses as a substrate supplemented with urea at 150rpm. Yeast cells were then separated by centrifugation and ruptured and autolysis was observed to be the most feasible method. Among various method employed to dry the yeast extract, spray dryer appeared as most efficient one. Yeast extract obtained after drying was subjected to different analyses and compared with commercial yeast extract. The produced yeast extract was applied in media preparation to grow different microorganisms including yeast, bacteria and fungi and considerable growth was observed. These results indicated that the developed process is a cost effective alternate approach for the production of yeast extract.

Nowadays, probiotics has been proposed for substituting antibiotics in animal feed, since the European Union (EU) banned the latter compounds in 2006 to avoid their serious side effects on human health. Therefore, this work aimed to produce a probiotic product by fed-fermentation of whey with kefir grains for use in animal feed. The whey was fermented with a combination of kefir grains AGK1 and the free biomass present in whole milk fermented milk used to activate these kefir grains. The probiotic culture obtained was characterized with high levels of biomass, total viability and antibacterial activity. Some probiotic properties of the probiotic culture were investigated in vitro, including its survival at low pH values, under simulated gastrointestinal conditions, after freezing in skim milk at − 20 ºC, and in the commercial feed during storage at room temperature. The viable cells of lactic and acetic acid bacteria and yeasts exhibited higher tolerance to acidic pH and simulated gastrointestinal conditions when the cells were protected with skim milk and piglet feed, compared with washed cells. The results indicated the feasibility of producing a probiotic product at a low cost with a potential application in animal feed.

The study of the impact of polyQ expansion driven proteotoxicity on cell metabolism has evi-denced NAD molecule as a central player in regulating metabolic response and cell repair in neurodegeneration. The decline in NAD intracellular levels represents a hallmark of ageing aris-ing from a dysregulation between NAD-consuming and biosynthetic enzymatic reactions in which NMNAT plays a key role. By an integrated bioanalytical and proteomic experimental design, the derangement of NAD metabolism deriving from the accumulation of polyQ-Htt protein has been analysed in Saccha-romices cerevisiae harbouring a 103Q-Htt and compared with wild-type Htt, in absence and in presence of a phenolic extract from EVOO, as a source of antioxidant bioactive compounds. Quantification of NAD and its precursors and proteomic analysis of the differential protein ex-pression has been performed by LC-DAD-MS. Chronological lifespan and NAD homeostasis was strongly impaired by proteotoxic polyQ-Htt, eliciting a depletion of NAD levels and accumulation of NaMN. In parallel a downregulation of Sirt2 expression levels has been observed. Phenolic extract administration had no meaningful ef-fects on NAD replenishment to prevent proteotoxicity. NaMN can represent a biomarker of accel-erated aging, and/or a signaling molecule for cell repair.

Yeasts are gaining increasing attention for their potential health benefits as probiotics in recent years. Researchers are actively searching for new yeast strains with probiotic properties (i.e, Debaryomyces hansenii; Kluyveromyces marxianus; Yarrowia lipolytica; Pichia hudriavzevii; Torulaspora delbrueckii) from various sources, including traditional fermented foods, the human gut, and the environment. This exploration is expanding the pool of potential probiotic yeasts beyond the well-studied Saccharomyces boulardii. Research suggests that specific yeast strains possess properties that could be beneficial for managing conditions like inflammatory bowel disease, irritable bowel syndrome, skin disorders and allergy. Additionally, probiotic yeasts may compete with pathogenic bacteria for adhesion sites and nutrients, thereby inhibiting their growth and colonization. They might also produce antimicrobial compounds that di-rectly eliminate harmful bacteria. To achieve these goals, the approach that uses probiotics for human health is changing. Next-generation yeast probiotics are emerging as a powerful new approach in the field of live biotherapeutics. By using genetic engineering, scientists are able to give these tools specialized capabilities. However, most research on these probiotic yeasts is still in its early stages, and more clinical trials are needed to confirm their efficacy and safety for various health condi-tions. This review could provide a brief overview of the situation in this field.

The intestinal microbiota and its functions are regarded as critical for host health and disease. Probiotics can influence the gut microbiome and its interactions with the host, and are currently defined as “live microorganisms that, when administered in adequate amounts, confer a health benefit on the host”. Probiotics have become common components of strategies to promote livestock health, welfare and productivity, not least due to restrictions on the use of antimicrobial drugs. Common probiotic organisms are considered commensals and are ‘generally recognized as safe’ (GRAS) via oral administration. This review outlines potential probiotic mechanisms, including recent findings. These mechanisms include those interactions primarily occurring between the supplemented probiotic microorganisms and the indigenous intestinal microbiota, perhaps within the gut lumen, as well as more direct interactions with the host via mucosal receptors or more distally following absorption of microbial components. There is good evidence that the gut microbiome is relatively stable in ‘healthy’ individuals and resistant to ‘colonisation’ by exogenous microbes, which helps exclude pathogens, but has implications for the establishment of probiotics, and could increase the importance of microbe-microbe interactions. However, such microbiomes may be receptive to complementary microbes or functions, while supplemented probiotics may dominate luminal populations, particularly in less populated regions of the intestine. Moreover, host-adapted microbes or microbiomes may elicit different host responses and/or be more effective. Some considerations for the interpretation of study results, including extrapolation from different models or microbial strains, are also included. In addition, notable mechanistic and/or pathogen challenge studies from pigs and poultry are highlighted to underline the recognised potential of probiotics in these species, particularly as the appropriate selection of microorganisms and their application continues to be better understood and improve.

Malassezia are lipid-dependent basidiomycetous yeast of the normal skin microbiome, although Malassezia DNA has been recently detected in other body sites and has been associated with cer-tain chronic human diseases. This new perspective raises many questions. Are these yeasts truly present in the investigated body site or were they contaminated by other body sites, adjacent or not? Does this DNA contamination come from living or dead yeast? If these yeasts are alive, do they belong to the resident mycobiota or are they transient colonizers which are not permanently established within these niches? And, finally, are these yeasts associated with certain chronic diseases or not? In an attempt to shed light on this knowledge gap, we critically re-viewed the 31 published studies focusing on the association of Malassezia spp. with chronic human diseases, including psoriasis, atopic dermatitis (AD), chronic rhinosinusitis (CRS), asthma, cystic fibrosis (CF), HIV infection, inflammatory bowel disease (IBD), colorectal cancer (CRC), and neurodegenerative diseases.

Marine yeasts have versatile applications in industrial, medical and environmental fields, but have received little attention compared to terrestrial yeasts and filamentous fungi. In this study, a phylogenetic analysis of 11 marine-derived yeasts was conducted using internal transcribed spacers and nuclear large subunit rDNA, and their bioactivities, such as antioxidant, antibacterial, and tyrosinase inhibition activities, were investigated. The 11 marine-derived yeasts were identified to belong to seven species including Geotrichum candidum, Metschnikowia bicuspidata, Papiliotrema fonsecae, Rhodotorula mucilaginosa, Vishniacozyma carnescens, Yamadazyma olivae, and Yarrowia lipolytica, and three strains of these were candidates for new species of the genera Aureobasidium, Rhodotorula, and Vishniacozyma. Most extracts showed antioxidant activity, whereas seven strains exhibited antibacterial activities against Bacillus subtilis. Only Aureobasidium sp. US-Sd3 among the 11 isolates showed tyrosinase inhibition. Metschnikowia bicuspidata BP-Up1 and Yamadazyma olivae K2-6 showed notable radical-scavenging activity, which has not been reported previously. Among the isolates, Aureobasidium sp. US-Sd3 exhibited the highest antibacterial and tyrosinase inhibitory activities. Overall, our results demonstrate the potential of marine-derived yeasts as a source of bioactive compounds for improving industrial applications.

The selectivity in selective autophagy pathways is achieved via the selective autophagy receptors (SARs) – proteins that bind a ligand on the substrate to be degraded and the Atg8-family protein on the growing autophagic membrane, phagophore, effectively bridging them. In mammals, the most common ligand of SARs is ubiquitin, a small protein modifier that tags substrates for their preferential degradation by autophagy. Consequently, the most common SARs are the ubiquitin-binding SARs, such as SQSTM1/p62 (sequestosome 1). Surprisingly, there is only one SAR of this type in yeast – Cue5, which acts as a receptor for aggrephagy and proteaphagy – pathways that remove the ubiquitinated protein aggregates and proteasomes, respectively. However, recent studies described the ubiquitin-dependent autophagic pathways that do not require Cue5, e.g. stationary phase lipophagy for intracellular lipid droplets and nitrogen starvation-induced mitophagy for mitochondria. What is the role of ubiquitin in these pathways? Here, we propose that the ubiquitinated lipid droplets and mitochondria are recognized by the alternative ubiquitin-binding SARs. Our analysis identifies the proteins that could potentially fulfill this role in yeast. We think that matching of the ubiquitin-dependent (but Cue5-independent) autophagic pathways with the ubiquitin-and-Atg8-binding proteins enlisted here might uncover the novel ubiquitin-binding SARs in yeast.

The principal goal of this current study was to isolate a new potential probiotic strain to prepare contamination-free probiotic dahi in a dosage form with desired shelf life. To achieve our target, firstly, probiotic dahi was prepared by a probiotic strain, Lactobacillus acidophilus JB2CON (OM909067) (identified by 16S sequencing) isolated from natural dahi in Dhaka, Bangladesh. The isolate showed five probiotic features by in vitro test and had an antagonistic effect against five different pathogens. More interestingly, it inhibited Escherichia coli (diameter of zone of inhibition = 15.75±0.35) the most. In the time-kill analysis method, E. coli was killed totally within 5.50 hours of fermentation. Finally, the time duration of 14 days was an ideal shelf life, because the probiotic count increased but a slight change in pH did not affect the taste (sourness/acidity) of dahi. The most significant impact of this research is that the probiotic dahi of our experiment might be used as a cheap option for preventing pathogenic Escherichia coli-related foodborne diseases worldwide, especially in underdeveloped countries like Bangladesh.

The use of Lactic Acid Bacteria (LAB) for probiotic enrichment of minimally processed fruit is a well-established practice in literature. In addition, several LAB demonstrated a strain-specific ability to control harmful microorganisms and decay agents, improving shelf-life, maintaining quality and promoting the safety of fruits and vegetables. Edible coatings can help modulate phenomena of gas exchange and water loss by fruits, representing protection from physical damage and spoilage phenomena linked to oxidation and the development of undesired microorganisms. At the same time, the coating can represent an innovative delivery matrix for the LAB strains of potential interest to improve safety and quality in the postharvest management of fruits. In this work, five Lactiplantibacillus plantarum strains, previously characterized for their probiotic and antifungal activity, have been incorporated into a sodium alginate coating to develop edible probiotic coatings with antifungal properties for table grapes cv. Italia. The transfer of probiotics and their survival were evaluated by comparing coated and uncoated table grapes during 14 days of cold storage at 4 °C. The alginate edible coating increased the number of probiotics transferred to the surface of the berries from about 5 to more than 7 Log CFU/g, with a crucial impact on the potential functional attributes of the final product. The ability of the functionalised coatings to counteract the decay development was evaluated on table grapes berries artificially contaminated with Aspergillus niger CECT 2805. A significant reduction of lesion diameter was observed in alginate coating with L. plantarum 11-A. The lesion diameter reduction was also accompanied by the reduction of the symptoms of infection, such as browning around the wound. These results suggest the application of selected strains of L. plantarum as promising bio-resources to enhance the overall value of ready-to-eat fruits and vegetables, particularly in combination with edible coating as a carrier matrix. While a strain-dependent effect was not detected with respect to the improvement of the number of cells in the edible coating, a variability depending on the biotype used was detected for the properties linked to biocontrol, suggesting that the inclusion in edible packaging may represent an innovative criterion in the selection of lactobacilli to be applied postharvest.

Whey proteins have excellent nutritional characteristics due to their levels of essential amino acids with high bioavailability. However, it has a high biochemical oxygen demand (BOD) and a considerable polluting potential, thus food manufacturers have opted to add whey to food formulations. The demand for beverages containing vitamins, probiotics, prebiotics, minerals, and bioactive compounds (antioxidants) with health benefits has increased and driven market growth. Therefore, this study aimed to develop a probiotic functional carbonated beverage from cheese whey and evaluate its microbiological, and physicochemical characteristics soon after the production and during storage. The viability and stability of probiotic, the microbiological characteristics, titratable acidity and sedimentation of the beverage were monitored during refrigerated storage for a month. The probiotic to be added to the formulation was established in a preliminary step. The production of this beverage proved to be a simple technology and the product was suitable for incorporation of the probiotic Bifidobacterium animalis subsp. lactis. The probiotic showed good viability and stability during storage. The microbiological quality of the beverage met the Brazilian legal standards. The pH and titratable acidity of the probiotic carbonated beverage remained stable during storage, and slight sedimentation was observed after one week of refrigerated storage.

<I>Saccharomyces cerevisiae</I> proliferates by budding, which includes the formation of a cytoplasmic protrusion called the ‘bud’, into which DNA, RNA, proteins, organelles, and other materials are transported. The transport of organelles into the growing bud must be strictly regulated for proper inheritance of organelles by daughter cells. In yeast, E3 ubiquitin ligases are involved in inheritance of mitochondria, vacuoles, and peroxisomes. These organelles are transported toward the tip of the growing bud by the myosin motor protein Myo2 along actin filaments. During organelle transport, adaptor proteins bridge the organelles and myosin. After reaching the bud, the adaptor proteins are ubiquitinated by E3 ubiquitin ligases and then degraded by the proteasome. Targeted degradation of the adaptor proteins is necessary to unload to unload the organelles from the actin–myosin machinery. Impairment of the ubiquitination of adaptor proteins results in a failure of organelle release from myosin, which in turn leads to abnormal dynamics, morphology, and function of the inherited organelles. Here, we summarize the role and regulation of E3 ubiquitin ligases during organelle inheritance in yeast.

Genetic switches can be utilized for many purposes in synthetic biology including the assembly of complex genetic circuits to achieve sophisticated cellular systems and the construction of biosensors for real-time monitoring of intracellular metabolite concentrations. Although genetic switches have mainly been developed in prokaryotes to date, eukaryotic genetic switches are increasingly being reported as both rational and irrational engineering technologies mature. In this review, we describe genetic switches in yeast based on synthetic transcription factors and/or synthetic promoters. We also discuss directed evolution technologies for the rapid and robust construction of yeast genetic switches.

Society considers wine as a special product among food and beverages because of its high gastronomical value and its positively distinctive quality. In recent years, philosophies of the agricultural techniques and development of the oenological technology have been focused on the reduction of wasteful, "polluting" elements, and trends are moving towards an environmental friendly approach. Due to the stricter regulations and rules (with the limited amount and selection of the permitted chemicals) resistant, also known as interspecific or innovative grape varieties can be the ideal basic materials of alternative cultivation technologies. In terms of variety selection, innovative varieties can be equivalent to international varieties, although organically their quality could not compete with them. These grapes are more resistant to various fungal diseases and infections than international varieties. Well-founded analytical and organoleptic results have to provide the scientific background of resistant varieties, as these cultivars with the environmental friendly cultivation techniques, could be the raw material of the future.

DNA is stored in the nucleus of a cell in a folded state; however, only the necessary genetic information is extracted from the required group of genes. The key to extracting genetic information is chromatin ambivalence. Depending on the chromosomal region, chromatin is characterized into low-density "euchromatin" and high-density "heterochromatin", with various factors being involved in its regulation. Here, we focus on chromatin regulation and gene expression by the yeast FACT complex, which functions in both euchromatin and heterochromatin. FACT is known as a histone H2A/H2B chaperone and initially reported as an elongation factor associated with RNA polymerase II. In budding yeast, FACT activates promoter chromatin by interacting with the transcriptional activators SBF/MBF via the regulation of G1/S cell cycle genes. In fission yeast, FACT plays an important role in the formation of higher-order chromatin structures and transcriptional repression by binding to Swi6, an HP1 family protein, at heterochromatin. This FACT property, which refers to the alternate chromatin-regulation depending on the binding partner, is an interesting phenomenon. Further analysis of nucleosome regulation within heterochromatin is expected in future studies.

Wound healing is a multi-factorial response to tissue injury, aiming to restore tissue continuity. Numerous recent experimental and some clinical studies, clearly indicate that probiotics applied topically promote the wound healing process. However, the precise mechanism by which they contribute to healing is not yet clear. Each strain appears to exert a distinctive, even multi-factorial action on different phases of the healing process. Given that a multi-probiotic formula exerts better results than a single strain, the pharmaceutical industry has embarked on a race for the production of a formulation containing a combination of probiotics, capable of playing a role in all the phases of the healing process. Hence, the object of this review is to describe what is known to date of the distinctive mechanisms of each of the most studied probiotic strains in order to further facilitate research towards the development of combinations of strains and doses, covering the whole spectrum of healing. Eleven probiotic species have been analyzed, the only criterion of inclusion being a minimum of two published research articles.

Findings on diet-health relationships have led many people to include healthier food items (including snacks) in their diets, e.g. those containing probiotic microorganisms. The purpose of this study was to compare two methods to produce probiotic freeze-dried banana slices - one of them consisting of impregnating slices with a suspension of probiotic Bacillus coagulans, the other based on coating the slices with a starch dispersion containing the bacteria. Both processes have resulted in viable cell counts above 7 log ufc.g-1, although the presence of the starch coating has avoided a significant loss of viability on freeze-drying. The coated slices were less crispy than the impregnated ones, according to the shear force test results. However, the sensory panel (with more than 100 panelists) have not perceived significantly texture differences. Both methods have presented good results, in terms of probiotic cell viability and sensory acceptability (the coated slices having been significantly more accepted than the non-probiotic control slices).

The aim of this paper was to test isolated and pre-selected bacteria strains from Nile tilapia intestinal mucus, collected in Brazilian commercial fish farming, to use as probiotics in the feeding. The variable were growth performance, hematology, microbiology and immunology. These strains were submitted to in vitro inhibition tests of pathogenic bacteria. Then, they were identified by PCR and sequencing, and comparing using the BLAST software. The experiment was carried out in the Fisheries Institute, Sao Paulo, Brazil, for 84 days, following the randomized experiment design with seven treatments and three replicates (21 experimental units): T0 – control, T1 – Bacillus velezensis, T2 – Bacillus pumilus, T3 – Bacillus subtilis, T4 –Enterococcus hirae T5 – Enterococcus faecium and T6 – mix of bacteria strains used to this experiment. The variables analyzed were: growth performance, DGGE (Denaturing gradient gel electrophoresis), phagocytes respiratory burst activity, hematology and bromatology. As result, the bacteria strains B. pumilus (T2) and all bacteria mix (T6), included in Nile tilapia diet, promoted better biomass values, with similar feed conversion rate of control group (p<0,05). Apparently, the microorganisms use does not affect the health and well-fare of the fish. The hematology index, except basophils and thrombocytes numbers do not demonstrated differences among the groups, as well as glycemia, lactate level, respiratory burst and lysozyme activities. In conclusion, the isolated and pre-selected bacteria from intestinal mucus of Nile tilapia, B. pumilus (T2) and mix (T6 - B. velezensis, B. pumilus, B. subtilis, E. hirae and E. faecium) improve the growth performance and may be used as probiotics for Nile tilapia.

Antibiotics are commonly used to treat infectious diseases. However, massive and inappropriate antibiotics usage cause many problems including the emergence and spread of antibiotic-resistant bacteria. To avoid this issue, in modern countries the interest of using probiotics in feed supplementation to promote health and prevent or treat intestinal infectious diseases in companion animals like dogs has been increasing. We evaluate the probiotic potential of Lactobacilli isolated from healthy dogs faeces. The isolated Lactobacilli were first confirmed by 16SrRNA sequencing, then in vitro tests were conducted to assess survival potential of Lactobacilli under simulated gastrointestinal conditions and adhesion ability to gut epithelia, effects on epithelial barrier function, anti-inflammatory activities, effects on defensin peptides (beta-defensin 3) and inhibitory effects on common pathogens. Lactobacilli showed considerable potential to survive in simulated gastrointestinal environmental conditions, low pH, high bile salt concentrations along with good adhesion properties with MODE-K cells. Pathogenic bacterial growth and their adhesion to MODE-K cells was significantly inhibited by Lactobacilli. Real-time PCR analyses further demonstrated that L. acidophilus strain AR1 and AR3 inhibit Salmonella-induced proinflammatory cytokines (IL-6, IL-8, 1ꞵ) production and reinforce expression of tight junction protein (occludin). None of the strain induce mRNA expression of beta-defensin 3 in MODE-K cells. Based on in vitro results the L. acidophilus strain AR1 has potential to be supplemented in canine feed. However, further in vivo studies investigating health-promoting effects are awaited.

The aim of the study was to assess the effect of the addition of inulin and the replacement of part of inulin with apple fiber on the physicochemical and organoleptic properties. Moreover, the survival of Bifidobacterium animalis ssp. Lactis Bb-12 and Lactobacillus rhamnosus was studied in sheep milk ice cream. There was no effect of apple fiber and the type of bacteria on the number of bacteria of the probiotics after fermentation. As a result of freezing, mixture containing Bifidobacterium animalis ssp. Lactis Bb-12, there was a significant reduction in the bacterial from 0.39 log cfu g −1 to 0.46 log cfu g −1. In all ice cream on the 21st day of storage, it exceeded 10 log cfu g –1, which means that the ice cream retained the status of probiotic products. The Lactobacillus rhamnosus ice cream showed a lower yellow colour compared to the Bifidobacterium Bb-12 ice cream. The overrun of sheep's milk ice cream was within a range from 78.50% to 80.41%. The appearance of sheep's milk ice cream is influenced considerably by the addition of fiber and the type of bacteria and the interaction between the type of bacteria and the addition of fiber and storage time and fiber.

Microbe-plant interactions are linked with the core microbiota, and both the plant and the microbial partners depend on one other to thrive in nature. However, why and how the below-ground core microbiota become established aboveground is poorly understood. We tracked the movement of a probiotic Streptomyces endophyte throughout a managed strawberry ecosystem. Probiotics in the rhizosphere and anthosphere were genetically identical, yet these niches were segregated in space and time. The probiotic in the rhizosphere moved upward via the vascular bundle, relocated to aboveground plant parts, and protected against Botrytis cinerea. It also moved from flowers to roots, and among flowers via pollinators that were protected against pollinator pathogens. Our results reveal a solid evidence in tripartite interaction with Streptomyces exploiting plant and pollinator partners.

In the last decades, technological advances related to RNA manipulation enabled and expanded its application in vaccine development. This approach comprises synthetic single-stranded mRNA molecules that direct the translation of the antigen responsible for activating the desired immune response. The success of RNA vaccines depends on the delivery vehicle employed. Among the systems, yeasts emerge as a new approach to a natural delivery platform. The presence of β-glucans and mannans in its wall is responsible for the adjuvant action of this system. Yeasts are already employed to deliver protein antigens, with success and efficacy demonstrated through pre-clinical and clinical trials. Yeast β-glucan capsules, microparticles, and nanoparticles are capable of modulating host immune responses and have a high capacity to carry RNA and small molecules, with bioavailability upon oral immunization and with targeting to receptors present in anti-gen-presenting cells (APCs). Besides, yeasts are interesting vehicles for the protection and specific delivery of therapeutic vaccines based on shRNA or dsRNA. In this review, we present an overview of the attributes of yeast or its derivatives for the delivery of RNA-based vaccines, discussing their current challenges and prospects for using this promising strategy.

Reversed Flow Gas Chromatography (R.F.G.C.) was employed to assess the impact of genetic modification on Saccharomyces cerevisiae yeast strains during the process of alcoholic fermentation, utilizing fig syrup. Multiple fermentations were carried out at various temperatures to evaluate the influence of genetic modifications on yeast strain efficiency. The study involved a wild-type yeast strain, W303, as a control, and two genetically modified strains, W_M4_533 and W_M4_558, sharing the same genetic background as the wild type. Notably, the genetic modifications in the Msn4p transcription factor involved the substitution of serine residues with alanine at positions 533 and 558, resulting in the development of psychrophilic or ethanol-resistant strains. Utilizing the R.F.G.C. method enabled the differentiation of the duration of alcoholic fermentation phases, providing insights correlated to the yeast cell life cycle. The values of rate constants (k) for each phase, conducted with both wild-type and genetically modified cells using RFGC, aligned with existing literature. Additionally, the calculation of activation energies for distinct phases revealed lower values for genetically modified strains compared to wild-type strains. This decrease in activation energies suggests enhanced efficiency in the alcoholic fermentation process for the genetically modified strains.

This study provides a comprehensive investigation of lactic acid bacteria (LAB) isolated from Argentinean Capsicum annum L. The research covers important aspects, including genotypic characterization, bacterial stress tolerance, adhesion ability, safety evaluation, and functional and technological properties. The predominant isolates were identified as Lactilactobacillus curvatus and Lactiplantibacillus plantarum. Rep-PCR analysis grouped the isolates into 11 clonal groups. Lp. plantarum LVP 40 and LV 46; Levilactobacillus brevis LVP 41, Pediococcus pentosaseus LV P43, and Lt. curvatus LVP44, displayed both safety and resilience against adverse conditions such a slow pH, bile, and simulated gastric and intestinal juices. Moreover, the LAB strains exhibited high hydrophobicity and auto-aggregation percentages, NaCl tolerance, and substantial acidifying capacity. LAB supernatants demonstrated promising surfactant and emulsifying properties. Likewise, they differentially inhibited Staphylococcus aureus and Pseudomonas aeruginosa biofilms, showcasing their potential as antipathogenic agents. Noteworthy some strains displayed considerable co-aggregation with these pathogens, and several isolates showed an effective antimutagenic and detoxifying power, further emphasizing their multifaceted capabilities. Five pepper bacterial strains showcased probiotic properties, suggesting their potential for gut health enhancement. In summary, these LAB strains hold promise as vegetable fermentation starters, contributing to food safety and versatile applications in food science.

The regulation of intracellular pH in yeast (Saccharomyces cerevisiae) cells is critical for cell function and viability. In yeast, protons (H+) can be excreted from cell by the plasma membrane ATPase Pma1 and pumped into vacuoles by the vacuolar H+-ATPase. Because Pma1 is critical to the survival of yeast cells, it is unknown whether other compensatory components are involved in the pH homeostasis in the absence of Pma1. To elucidate how intracellular pH is regulated independently of Pma1, we employed a screening approach by exposing the yeast haploid deletion mutant library (ver 4.0) to the selective plant plasma membrane H+-ATPase inhibitor PS-1, which we previously reported. After repeated screenings and verification, we identified two proteins, ALY1 and ALY2, that play a role in the regulation of intracellular pH when Pma1 is deficient. Our research uncovers a new perspective on the regulation of intracellular pH related to Pma1 and also preliminarily reveals a role for ALY1 and ALY2 in the regulation of intracellular pH.

A perfectly balanced wine can be said to create a symphony in the mouth. To achieve the sublime, both in wine and music, requires imagination and skilled orchestration of artistic craftmanship. For wine, inventiveness starts in the vineyard. Similar to a composer of music, the grapegrower produces grapes through a multitude of specifications to achieve a quality result. Different Vitis vinifera grape varieties allow the creation of wine of different genres. Akin to a conductor of music, the winemaker decides what genre to create and considers resources required to realise the grape’s potential. A primary consideration is the yeast: inoculate the grape juice or leave it ‘wild’; which specific or combined Saccharomyces strain(s) should be used; or proceed with a non-Saccharomyces species? Whilst the various Saccharomyces and non-Saccharomyces yeasts perform their role during fermentation, the performance is not over until the ‘fat lady’ (S. cerevisiae) has sung (i.e. the grape sugar has been fermented to specified dryness and alcoholic fermentation is complete). Is the wine harmonious or discordant? Will the consumer demand an encore and make a repeat purchase? Understanding consumer needs lets winemakers orchestrate different symphonies (i.e. wine styles) using single- or multi-species ferments. Some consumers will choose the sounds of a philharmonic orchestra comprising a great range of diverse instrumentalists (as is the case with wine created from spontaneous fermentation); some will prefer to listen to a smaller ensemble (analogous to wine produced by a selected group of non-Saccharomyces and Saccharomyces yeast); and others will favour the well-known and reliable superstar soprano (i.e. S. cerevisiae). But what if a digital music synthesiser ‒ such as a synthetic yeast ‒ becomes available that can produce any music genre with the purest of sounds by the touch of a few buttons? Will synthesisers spoil the character of the music and lead to the loss of the much-lauded romantic mystique? Or will music synthesisers support composers and conductors to create novel compositions and even higher quality performances that will thrill audiences? This article explores these and other relevant questions in the context of winemaking and the role that yeast and its genomics play in the betterment of wine quality.

The goal of the current study was to discover a novel potential probiotic strain of Lactobacillus spp. with anti-Escherichia coli activity from locally produced yogurt in Tongi, Gazipur, Bangladesh, compare its antagonistic activity with a commercial probiotic mixture of several strains, and approve a novel method for confirming the viability and relative abundance of the microbial community in a probiotic mixer. We carried out 16S sequencing, 16S metagenomics, Transmission Electron Microscopy (TEM) analysis, and other in vitro laboratory experiments to reach this objective. The strain TY-11 was identified as Lactobacillus delbrueckii subsp. indicus (16S sequence accession number OQ652026). It was gram-positive, anaerobic, lactose fermenting, and round-ended rod that typically measured 0.7 to 1.3 µm by 2.2 to 9 µm. In addition to having seven probiotic characteristics, it also showed an antagonistic impact on six different pathogens, but what's more noteworthy is that E. coli was the pathogen it inhibited most strongly (inhibition zone diameter was 18.88±0.18 mm). The most important and ground-breaking finding of this work was determining the probiotic features of a new probiotic strain, TY-11, whose antibacterial activity was virtually as effective as that of the probiotic combination with three different strains. Furthermore, the results of 16S high throughput sequencing and the conventional plate count method demonstrated a strong correlation (0.999) at the genus level, indicating that the use of both of these approaches in combination may be a practical way to assess the relative abundance of the microbial community and their viability in commercially available probiotic blends.

Bovine whey cheese (WC) is a product from southern European countries that presents some challenges: their elaboration process involves high energy inputs, the yield is low and WC has a short shelf-life. The application of ultrafiltration to bovine whey before its manufacture and the employment of protective cultures can reduce these disadvantages. The aim of this work was to manufacture whey cheeses using ultrafiltrated (UF) concentrated bovine cheese whey with added probiotics or probiotics plus protective cultures. Three types of WC were produced: control, without any addition (C); with the addition of the probiotic Lactobacillus acidophilus (LA5); with the addition of Lactobacillus acidophilus plus a protective culture containing Lactobacillus paracasei and Lactobacillus rhamnosus (LA5FQ4). The WCs were stored under refrigerated conditions for 28 days. The products with added cultures presented a lower pH and a higher titratable acidity when compared to the control. Sample LA5 presented the lowest pH and the highest titratable acidity while LA5FQ4 presented intermediate values. Slight differences were observed between products regarding color parameters, chiefly as a result of storage time. The samples with added cultures were firmer when compared to the control, showing LA5 cheeses the highest values at the end of the storage. Lactobacilli and lactococci counts were of the order of log 8-9 CFU/g for the products with added cultures. Lower levels of yeasts and moulds were detected on the sample with the protective culture (LA5FQ4), that by the end of storage presented counts one log cycle lower than C and LA5. Hence, it is evident the beneficial impact of the protective culture on the shelf life of the product. In what concerns to sensory evaluation, LA5FQ4 cheeses obtained the highest scores regarding all parameters evaluated. It could be concluded that the use of UF associated to the addition of protective cultures can be very useful to reduce the energy consumption of the process, prolong the shelf life of WC, and to improve its sensory properties.

A selection of 36 commercial probiotic fermented dairy products from UK and Europe markets were evaluated for the numbers, types and viability of Lactobacillus strains against the stated information on their packages. A comparative study was carried out on selectivity of MRS-Clindamycin, MRS-Sorbitol and MRS-IM Maltose, to select the right medium for enumeration of probiotic Lactobacillus. Based on selectivity of medium for recovery of the targeted lactobacilli and also simplicity of preparation, MRS-Clindamycin was chosen as the best medium for enumeration of probiotic Lactobacillus in fermented milks. The results of enumeration of lactobacilli showed that 22 out of a total 36 tested products contained more than 106 colony forming units/g at the end of their shelf- life, which comply with the recommended minimum therapeutic level for probiotics. Rep-PCR using primer GTG-5 was applied for initial discrimination of isolated strains, and isolates, which presented different band profile, were placed in different groups. The isolated Lactobacillus spp. were identified mainly as Lactobacillus acidophilus, Lactobacillus casei and Lactobacillus paracasei by analysis of partial sequences of the 16S ribosomal RNA and rpoA genes. In conclusion, it is unknown to recommend the adequate number of probiotic bacteria to be consumed to ensure the beneficial properties.

This paper reports for the first time on the production of probiotic alcoholic and non-alcoholic rosé wines with enhanced health benefits made with Saccharomyces cerevisiae var. boulardii probiotic yeast. The alcohol, sugar, volatile acidity lactic and malic acid contents were assessed for S. cerevisiae var. boulardii before and after fermentation and distillation and compared with a conventional Saccharomyces cerevisiae (ex-bayanus) yeast. The free amino nitrogen and gluconic acid concentrations in the musts were determined. Yeast viability was evaluated after fermentation and distillation as a function of time (0, 15 days, 3 months and 6 months) both at room temperature (25±0.5ºC) and refrigerator temperature (4±0.5ºC). The results obtained showed that the probiotic rosé wine produced with S. cerevisiae var. boulardii possesses the typical values and sensory attributes of other commercial wines produced with S. cerevisiae (ex-bayanus). The probiotic S. cerevisiae var. boulardii yeast survives the high alcohol content produced during fermentation and vacuum distillation. The study also showed that this probiotic rosé wine stored either at room temperature or in a refrigerator keeps its probiotic viability for at least six months, which makes it a promising for large-scale production, in which long storage times are required by both producers and consumers.

Malassezia is lipid-dependent commensal yeast of the human skin. The different culture media and skin sampling methods used to grow these fastidious yeasts are a source of heterogeneity in culture-based epidemiological study results. This study aimed to compare the performances of three methods of skin sampling, and two culture media for the detection of Malassezia yeasts by culture from the human skin. Three skin sampling methods, namely sterile gauze, dry swab and TranswabTM with transport medium, were applied on 10 healthy volunteers. Each sample was further inoculated onto either the novel FastFung medium or the reference Dixon agar for the detection of Malassezia spp. by culture. At least one colony of Malassezia spp. grew on 93/300 (31%) of the cultures, corresponding to 150 samplings. The positive culture rate was 67%, 18%, and 15% (P &lt; 10-3), for samples collected with sterile gauze, TranswabTM, and dry swab, respectively. The positive culture rate was 62% and 38% (P &lt; 0.003) by using the FastFung and the Dixon media, respectively. Our results showed that sterile gauze rubbing skin sampling followed by inoculation on FastFung medium should be implemented in the routine clinical laboratory procedure for Malassezia spp. cultivation.

LSM4 is an essential yeast gene encoding a component of different LSM complexes involved in the regulation of mRNA splicing, stability and translation. In previous papers we reported that the expression in S. cerevisiae of the K. lactis Lsm4 gene lacking the C-terminal Q/N-rich domain in a Lsm4 null strain S. cerevisiae (Sclsm4D1) restored cell viability. Nevertheless, in this transformed strain we observed some phenotypes which are typical markers of regulated cell death, Reactive Oxygen Species and oxidated RNA accumulation. In this paper we report that a similar truncation operated in the S. cerevisiae LSM4 gene confers to cells the same phenotypes observed with the K.lactis LSM4D1 gene. Up to now there was no evidence on the direct involvement of LSM4 in autophagy. Here we found that the Sclsm4D1 mutant showed defects in the induction of autophagy and it was very sensitive to nitrogen starvation or treatment with low doses of rapamycin, an inducer of autophagy. Moreover, both during nitrogen starvation and ageing, the Sclsm4D1 mutant accumulated cytoplasmic autophagy-related structures, suggesting a role of Lsm4 in Phagophore Assembly Site (PAS) processing and/or vacuolar autophagosome internalization

The increasing demand for craft beer drives the search for novel ale yeast cultures from brew-ing-related wild environments. The focus of bioprospecting for craft cultures is to identify feral yeasts suitable to imprint unique sensorial attributes to the final product. Here, we integrated phylogenetic, genotypic, genetic, and metabolomic techniques to demonstrate that sour beer during aging in wooden barrel is a source of suitable craft ale yeast candidates. Differently from traditional lambic beer maturation phase, during aging of sour-matured production style beer, different biotypes of Saccharomyces cerevisiae dominated the cultivable in-house mycobiota, fol-lowed by Pichia membranifaciens, Brettanomyces bruxellensis, and Brettanomyces anomalus. In addi-tion, three putative S. cerevisiae x Saccharomyces uvarum hybrids were identified. S. cerevisiae feral strains sporulated, produced viable monosporic progenies, and have STA1 gene as well as the full-length promoter. During hopped wort fermentation four S. cerevisiae strains and the S. cere-visiae x S. uvarum hybrid WY213 overcame non-Saccharomyces strains in fermentative rate and ethanol production, except for P. membranifaciens WY122. This strain consumed maltose after a long lag phase, in contrast with the phenotypic profile descripted for the species. According to STA1+ genotype S. cerevisiae partially consumed dextrin, but no 4-vynil guaiacol was detected among VOCs, suggesting that these strains were phenolic off flavor-negative (POF-). In conclu-sion, the strains characterized here have relevant brewing properties and are exploitable as in-digenous craft beer starters.

Whole-genome DNA sequencing of Lactobacillus D75 and D76 strains (Vitaflor, Russia) was performed using the PacBio RS II platform, followed by de novo assembly with SMRT Portal 2.3.0. The average nucleotide identity (ANI) test showed that both strains belong to the Lactobacillus helveticus, but not the L. acidophilus as previously assumed. 31 exopolysaccharide (EPS) production genes (nine of which form a single genetic cluster), 13 adhesion genes, 38 milk protein and 11 milk sugar utilization genes, 13 genes for and against specific antagonistic activity, aight antibiotic resistance genes, and also three CRISPR blocks and eight Cas I-B system genes were identified in the genomes of the both strains. The expression of some genes was confirmed. In fact, the presence of identified genes suggests that L. helveticus D75 and D76 are able to form biofilms on the outer mucin layer, inhibit the growth of pathogens and pathobionts, utilize milk substrates with the formation of digestible milk sugars and bioactive peptides, resist bacteriophages and show some genome-determined resistance to antibiotics, stimulate the host’s immune system. Pathogenicity genes have not been identified. The study results confirm the safety and high probiotic potential of the strains.

This study aimed to explore the fermentation behavior of selected potential probiotic strains during sorghum fermentation, considering their microbial counts, pH levels, and antibacterial activity, their ability to break down starch. The fermentation processes were conducted in the presence and absence of glucose, and the lactic acid bacterial strains underwent controlled fermentation to assess microbial counts, amylase activity, pH, and lactic acid levels in the presence and absence of glucose. In addition, the inhibitory activity of these potential probiotic strains against pathogenic bacteria (Salmonella typhimurium, Escherichia coli, and Staph-ylococcus aureus) was examined through co-culturing. The results demonstrated that all nine Lactobacillus strains exhibited robust growth in both sugar and sugar-free fermentation experiments. Glucose supple-mentation significantly enhanced lactic acid yield which was in a range of 0.19-0.52 % compared to fer-mentation without glucose with a range of 0.03-0.29 %. The selected Lactobacillus strains effectively lowered the pH to less than 4 after 24 hours, producing substantial lactic acid. Notably, in the absence of glucose, only Lb. helveticus D7 and Lb. amylolyticus D12 achieved pH levels below 4 after 8 hours, producing the highest lactic acid amounts of 0.27 and 0.29 % after 24 hours, respectively. Amylase activity was detected on two strains D7 and D12. Furthermore, most of the tested Lactobacillus strains demonstrated complete inhibition of pathogen growth after 24 hours of co-culturing, suggesting their potential for enhancing the safety quality of sorghum-based fermented products.

The continued exploration of the diversity of lactic acid bacteria in little-studied ecological niches represents a fundamental activity to understand the diffusion and biotechnological significance of this heterogeneous class of prokaryotes. In this study, Lactiplantibacillus plantarum (Lpb. plantarum) strains have been isolated from Tunisian vegetable sources, including fermented olive, fermented pepper and from dead locust intestines, subsequently evaluated for their antimicrobial activity against foodborne pathogenic bacteria, including Escherichia coli O157:H7 CECT 4267 and Listeria monocytogenes CECT 4031, as well as against some fungi, including Penicillium expansum, Aspergilus niger and Botrytis cinerea. In addition, their resistance to the oro-gastro-intestinal transit, aggregation capabilities, biofilm production capacity, adhesion to human enterocyte-like cells and cytotoxicity to colorectal adenocarcinoma cell line were determined. Further, the adhesion to tomatoes and the biocontrol potential on this model food matrix were analyzed. It has been found that all the strains were able to inhibit the indicator growth, mostly through organic acids production. Besides, these strains showed promising probiotic traits, including in vitro tolerance to oro-gastrointestinal conditions, adhesion to abiotic surface and to Caco-2 cells. Moreover, all tested Lpb. plantarum strains were able to adhere to tomatoes with similar rate (4.0-6.0 LogCFU/g tomato). The co-culture of LAB strains with pathogens on tomatoes showed that Lpb. plantarum could be a good candidate to control pathogen growth. Nonetheless, further studies are needed to guarantee their use as probiotic strains for biocontrol on food matrices.

Inorganic polyphosphate (polyP) is an important factor of stress tolerance in microbial cells. In yeast, the major enzyme of polyP biosynthesis is Vtc4, a subunit of the vacuole transporter chaperone (VTC) complex. In this study, we demonstrated that Vtc4 knockout in Saccharomyces cerevisiae not only decreased polyP content but also caused shifts in the composition of the intracellular polyP pool and changed the stress tolerance profile. In the mutant S. cerevisiae, the level of short-chain acid-soluble polyPs was decreased nearly 10-fold, whereas that of longer acid-insoluble polyPs was decreased only 2-fold, suggesting the existence of other enzymes compensating the production of long-chain polyPs. The Δvtc4 mutant showed inhibition of Mg2+-dependent phosphate uptake and decreased resistance to alkaline stress but increased tolerance to oxidation and heavy metal ions, especially Mn2+. Quantitative PCR revealed the upregulation of the DDR2 gene implicated in multiple stress responses and downregulation of PHO84 encoding a phosphate and Mn2+ transporter, which could account for the effects on phosphate uptake and Mn2+-related stress response in the Δvtc4 mutant. Our study indicates that short-chain polyPs, plays an important role in the regulation of stress response in yeast.

Exploring the dynamic behavior of cellular metabolism requires a standard laboratory method that guarantees rapid sampling and extraction of the cellular content. We propose a versatile sampling technique applicable to cells with different cell wall and cell membrane properties. The technique is based on irreversible electroporation with simultaneous quenching and extraction by using a microfluidic device. By application of electric pulses in the millisecond range, permanent lethal pores are formed in the cell membrane of Escherichia coli and Saccharomyces cerevisiae, facilitating the release of the cellular contents; here demonstrated by the measurement of glucose-6-phosphate and the activity of the enzyme glucose-6-phosphate dehydrogenase. The successful application of this device was demonstrated by pulsed electric field treatment in a flow-through configuration of the microfluidic chip in combination with sampling, inactivation, and extraction of the intracellular content in a few seconds. Minimum electric field strengths of 10 kV/cm for E. coli and 7.5 kV/cm for yeast S. cerevisiae were required for successful cell lysis. The results are discussed in the context of applications in industrial biotechnology, where metabolomics analyses are important.

Probiotics are namely yeast and bacteria which are regarded as dietary supplements and food. Many probiotics are being commonly used now, the most frequently used, lactic acid bacteria are called Lactobacillus and Bifidobacterium. Many of the diseases associated with GIT are related to microbiota found in GIT, an imbalance of which causes gastrointestinal diseases. Probiotics, in light of scientific literature, are thought to play an important role in relieving symptoms of many diseases associated with GIT, i.e. beneficially regulating the microbiota composition. In the present review, we aimed to highlight the main considerations for main probiotic formulations to date. This study reviews the role of different probiotic formulations introduced so far in treating GI diseases in cohort ageing ≤18 years. We searched PubMed and Clinicaltrials.gov without any restrictions. This study comprises the descriptive and comparative analysis between Single-organism and Composite probiotics. These GI diseases include NEC, FAP, AGE, Acute Diarrhea, Ulcerative Colitis and etc. The results have been categorized according to title and outcomes. The positive outcomes emphasize the drug’s effectiveness in improving health and the negative outcome elaborates on the adverse effects the drug may have shown. Conclusively, discussed practices will assist in reducing GI disorders and strengthening the gut. Further insight into the various gut microbes and microbiomes with specific demographic is recommended.

Mycoviruses are widespread across the major taxa of fungi including yeasts. Mycoviruses in the yeast fungus Geotrichum candidum are not well studied with three G. candidum-associated viral species characterized to date, all of which belong to the Totiviridae genus Totivirus. In this study, we report the molecular characteristics of another two totiviruses co-infecting isolate Gc6 of G. candidum. The two totiviruses were tentatively named Geotrichum candidum totivirus 2 (GcTV2-Gc6) and GcTV4-Gc6). Both viruses have typical genome organization of totiviruses comprising two ORFs encoding capsid protein (CP) and RNA-dependent RNA polymerase (RdRp) at the N and C termini, respectively. Genomes of GcTV2-Gc6 and GcTV4-Gc6 are 4592 and 4530 bp long, respectively. Both viruses contain the -1 frameshifting elements and their proteins could be expressed as a single fusion protein. GcTV2-Gc6 is closely related to a totivirus isolated from the same host whereas GcTV4-Gc6 is related to insect-associated totiviruses. The phylogenetic analysis indicated that GcTV2-Gc6 and GcTV4-Gc6 belong to two different sister clades, I-A and I-B, respectively. It is interesting that all viruses identified from G. candidum belong to the genus Totivirus. It is possible that the RNA interference (RNAi) mechanism is not active to suppress totivirus accumulation in isolate Gc6.

Candida albicans is a major fungal pathogen of humans, accounting for 15% of nosocomial infections with an estimated attributable mortality of 47%. C. albicans is usually a benign member of the human microbiome in healthy people. Under constant exposure to highly dynamic environmental cues in diverse host niches, C. albicans has successfully evolved to adapt to both commensal and pathogenic lifestyles. The ability of C. albicans to undergo a reversible morphological transition from yeast to filamentous forms is a well-established virulent trait. Over the past few decades, a significant amount of research has been carried out to understand the underlying regulatory mechanisms, signaling pathways, and transcription factors that govern the C. albicans yeast-to-hyphal transition. This review will summarize our current understanding of well-elucidated signal transduction pathways that activate C. albicans hyphal morphogenesis in response to various environmental cues and the cell cycle machinery involved in the subsequent regulation and maintenance of hyphal morphogenesis.

Introduction. The use of probiotics demonstrate efficacy against obesity and metabolic syndrome (MetS). Detection effective probiotic strains for hyperglycemia and immunity associated with is important task. The aim of the study was to evaluate an influence of Lactobacillus casei IMV B-7280 separately and composition L. casei IMV B-7280 / Bifidobacterium animalis VKB / B. animalis VKL on the levels of blood glucose and immunity in obese mice. Meterials and methods. Obesity was induced by fat-enriched diet (FED) in male BALB/c mice. Obese mice were transferred to standard diet and received per os probiotic strains daily during 10 days. We measured tumor necrosis factor-a (TNF-a) in blood serum using enzyme-linked immunosorbent assay and functional activity of peritoneal exudate macrophages (PEMs). Glucose levels in blood was defined with glucometer. Results. We ascertained that all probiotic strains induced reducing mice weight and visceral fat, normalization of TNF-a production and functional activity of PEMs. Treatment with L. casei IMV B-7280 was associated with decreasing blood glucose levels. No normalization of glucose and TNF-a levels in obese mice, transferred to standard diet without probiotic treatment, although we revealed decreasing their weight and visceral fat and partial recover of functional activity of PEMs. Conclusions. Probiotic strain L. casei IMV B-7280 (separately) and composition L. casei IMV B-7280 / B. animalis VKB / B. animalis VKL can re-equilibrate metabolic and inflammation indices in mouse obesity model. L. casei IMV B-7280 alone was more efficient in decreasing glucose levels than composition of strains.

Fermented rice is known as a healthy food due to the presence of lactic acid bacteria as fermentative microorganisms. The study was carried out to discover and characterize the lactic acid bacteria (LAB) from white and red fermented rice (BG varieties). Fermentation was carried out naturally by soaking red, white, raw and cooked rice in water using an earthen pot. Species were confirmed based on 16S r DNA gene sequencing and studied for phenotypic characteristics including morphological, physiological (growth temperature, salt tolerance, milk coagulation) and biochemical characteristics (carbohydrate fermentation pattern) using API 50CH kits. Distinct clusters like cocci (48), diplo cocci (30) and rods (30) shaped bacteria were observed from fermented rice. Five species of lactic acid bacteria including Latilactobacillus curvatus GR Lb1,2 10 and 11 (predominant Bacillus species), Latilactobacillus graminis Lb8, Limosilactobacillus fermentum GRLb17, Weissella confuse GRLb4 and Pediococcus pentosaceus GRLc1 were identified. The base pair length of amplified DNA for the isolates was 1500 Bp. The beneficial physiological and biochemical properties of isolated Lactobacillus species from fermented rice revealed their potential applications in the food industry.

Fermented food matrices, including beverages, can be defined as the result of the activity of complex microbial ecosystems where different microorganisms interact according to different biotic and abiotic factors. Certainly, in industrial production the technological processes aim to control the fermentation to place safe foods on the market. Therefore, if food safety is the essential prerogative, consumers are increasingly oriented towards a healthy and conscious diet driving the production and consequently the applied research towards natural processes. In this regard, the aim to guarantee safety, quality and diversify of products should be reached limiting or avoiding the addition of antimicrobials or synthetic additives using the biological approach. In this paper, following the recent re-evaluation of non-Saccharomyces yeasts (NSYs) has been re-viewed the bio-protectant and biocontrol activity with a particular focus on their antimicrobial power, probiotic features and promoting functional aspects. In this review, the authors would underline the contribution of NSYs in all food production chain and its role in the technological and fermentative features for their practical and useful use as biocontrol agent in food preparations.

Aquaculture is one of the fastest-growing animal food-producing agricultural industries in the world and proper performance of fish in morphological, physiological and immunological aspects is important for fish production and sustainable expansion of aquaculture. But several inhibitors like disease, pathogen, and adverse environment can overpower these performances. At present, antibiotics in preventing these inhibitors have been seen as becoming favorable to those inhibitors. So, Bacillus, an important group of probiotic bacteria can be an alternative to these antibiotics in aquaculture. Bacillus has been seen used in different experiments, mainly as a supplement in feed at various concentrations. Bacillus showed effective results like improved growth with minimum cost, improvement in reproduction, hematology, improved immune response and disease, and stress resistance as well as better proximate composition in different fish species. Application of Bacillus strains has proven efficient in improving water quality by reducing ammonia and nitrite toxicity, harmful algal blooms and utilization of H⁺ ion. Larger application of probiotic Bacillus instead of the hazardous synthetic chemicals would promote eco-friendly low-input sustainable aquaculture for food and nutritional security of the increasing world population. So many more experiments should be conducted in commercially important fishes for better growth and health of fishes which will certainly increase fish

The utilization of fermented foods has been a longstanding practice in human civilization worldwide for example sauerkraut from the Roman Empire, Jiangshui, and PaoCai, which are popular traditional foods in China. In Korea, Russia, and Mongolia, Kimchi and Kefir are widely used. Similarly, Japan, Indonesia, and Pakistan have their traditional fermented foods such as Miso, Natto, Tempeh, and sourdough, respectively. In America and Europe, fermented alcoholic beverages made from sorghum and maize are among the most common. Nevertheless, the full potential of fermented foods to enhance the bioavailability of bioactive compounds and restore probiotic communities has yet to be thoroughly explored. In this review, we focus on the bioactive compounds and probiotic stability in food fermented with probiotic bacterial strains. Probiotic fermented food improves the bioactive compound contents and has been gaining interest in basic and clinical research. Bioactive compounds, including phenolic, alkaloids, terpenoids, flavonoids, stilbenes, coumarins, tannins, anthocyanidins, flavones, isoflavonoids, and polyphenols, along with beneficial bacteria such as Lactobacilli, Bifidobacteria, Pediococcus, and Weissella demonstrate increased levels and restoration in probiotic fermented foods. These bioactive compounds, combined with a thriving microbiota, play a role in preventing viral infections by targeting influenza, noroviruses (NoVs), Murine norovirus-1 (MNV-1), and COVID-19, while also stimulating the host's immune function. Clinical and pre-clinical investigations are warranted to explore the dose-response and duration efficacy of probiotic fermented foods against viral infections.

This study aimed to exploring the intra-species distribution of genetic characters that favor the persistence in the gastrointestinal tract (GIT) and host interaction of bacteria belonging to the species Lacticaseibacillus genus. These bacterial species comprise commercial probiotics with the widest use among consumers and strains naturally occurring in GIT and in fermented food. Since little is known on the distribution of genetic traits for adhesion capacity, polysaccharide production, biofilm formation, utilization of substrates critically important for survival in GIT, that influence probiotic characteristics, a list of genetic determinants involved in such functions was created by a search for specific genes involved in the above aspects in the genome the extensively characterized probiotic L. rhamnosus GG. The presence/absence and variability of each gene in other Lacticaseibacillus spp. genomes was assessed by alignment with the publicly available fully annotated genome sequences. Eighty-two gene loci were compared, and 49 of these were found to be absent in some genomes in a species or strain-specific mode. A set of genes was found to be conserved, indicating that all strains of the genus may exert some probiotic effects. Among the variable loci a taurine utilization operon and a α-L-fucosidase were examined for presence/absence in 26 strains isolated from infant feces by PCR based tests. Results were variable among the isolates, though their common origin indicated the capacity to survive in the intestinal niche. This study indicated that the capacity to exert probiotic actions of Lacticaseibacillus spp. depends on a conserved set of genes and is enhanced by variable genetic factors whose role is only in part elucidated. The selection of strains of the most promising probiotic candidates to be used in food is feasible by analyzing presence/absence of a set of variable traits.

Probiotics make up a large and growing segment of the commercial market of dietary supplements and are touted as offering a variety of human health benefits. Some of the purported positive impacts of probiotics include, but are not limited to, stabilization of the gut microbiota, prevention of gastrointestinal disorders and modulation of the host immune system. Current research suggests that the immunomodulatory effects of probiotics are strain specific and vary in mode of action. Here, we examined the immunomodulatory properties of Bacillus subtilis strain DE111 in a healthy human population. In a randomized, double blind, placebo-controlled four-week intervention, we examined peripheral blood mononuclear cells (PBMCs) at basal levels pre- and post-treatment as well as in response to stimulation with bacterial lipopolysaccharide (LPS). We observed an anti-inflammatory effect of B. subtilis, manifested as a decrease in immune cell populations within the basal state along with an increase in anti-inflammatory immune cells in response to LPS stimulation. Overall gastrointestinal health, microbiota, and circulating and fecal markers of inflammation and gut barrier function were largely unaffected by DE111 treatment. These data suggest that the novel probiotic B. subtilis DE111 may have clinical applications in modulating immune homeostasis via anti-inflammatory mechanisms.

Cytokinesis, as the last stage of the cell division cycle, is a tightly controlled process amongst all eukaryotes, and defective division can lead to severe cellular consequences, and is implicated in serious human diseases and conditions such as cancer. Both mammalian cells and the fission yeast Schizosaccharomyces pombe use binary fission to divide into two equal sized daughter cells. In S. pombe such cytokinetic division is driven by the assembly of an actin-myosin contractile ring (ACR) at the cell equator between the two cell tips. The ACR is composed of a complex network of membrane scaffold proteins, actin filaments, myosin motors, and other cytokinesis regulators. Contraction of the ACR leads to the formation of a cleavage furrow which is severed leading to the final cell separation during the last stage of cytokinesis, abscission. This review describes recent findings defining the three phases to establish cytokinesis in S. pombe: ACR assembly, ACR constriction, and its coordination with septation. Collectively, we provide an overview of current understanding of the mechanisms regulating the ACR-mediated cytokinesis in S. pombe.

How to improve plant tolerance and yield under salt stress is critical for ensuring sufficient food supply since plant survival and agricultural productivity are both affected by salinity. Some evidence has showed that beneficial microorganisms have a high ability to improve plant salt tolerance and increase crop yield. But few studies were involved in effects of halotolerant yeasts on plants under salt stress. In this present research, Meyerozyma guilliermondii, a halotolerant yeast, was inoculated with tomato plants followed by salt treatment of four different NaCl concentrations (0, 100, 200, and 300 mM). Our results showed that inoculation of M. guilliermondii increased the chlorophyll biosynthesis and photosynthetic machinery effectiveness under salt stress, contributing to biomass accumulation. Under salt treatment of 300 mM NaCl, the yeast inoculation significantly increased ascorbate concentrations in leaves, yet showed no effects on levels of glutathione and proline. Antioxidant enzymes were affected differently by the yeast inoculation. It was found that the yeast inoculation increased superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activities under 300, 100, and 200 mM NaCl, respectively. Total soluble sugar levels increased in inoculated tomato plant leaves; however, there were no significant differences between different NaCl concentrations. Under 300 mM NaCl, the yeast inoculation significantly decreased H2O2 levels and reduced malondialdehyde levels. All together, our results showed that halotolerant yeast M. guilliermondii inoculation might be a strong candidate for regulating tomato growth under salt stress by increasing ability to scavenge reactive oxygen species and chlorophyll intactness, and by strengthening photosynthetic machinery.

The aim of this study was to assess the effect of added cultures of probiotic lactic acid bacteria on physicochemical, textural, and sensory characteristics of fresh cheese as well as their viability. Probiotic strains Lactobacillus acidophilus and L. rhamnosus were evaluated at two inoculation times (before the renneting and salting processes) following a factorial treatment arrangement. The bacterial count (log CFU g-1), acidity determination (% lactic acid), pH and percentage of syneresis were evaluated at 0, 1, 7, 14 and 21 days of refrigerated storage (4–5 °C). The texture profile analysis and an extended preference test were carried out at 21 days of storage. At the end of storage, fresh cheeses containing L. rhamnosus, added prior to the renneting process, had similar acidity values (0.18% lactic acid), pH (5.36) and percentage of syneresis (4.71%) compared to those for the cheeses without probiotics or control. Excluding adhesiveness, no differences were observed in texture profile parameters among the assessed cheeses. The fresh cheese supplemented with L. rhamnosus before the renneting process had a microorganism count greater than 6 logarithmic cycle/g, and the highest preference among treatments. The results from this study suggest that fresh cheese can be an excellent matrix for probiotic microorganisms, in particular L. rhamnosus.

A total of thirty-two aerobic spore former strains were isolated from intestinal samples of healthy children and analyzed for their hemolytic and antibiotic-resistance activities. Four strains selected as non-hemolytic and sensitive to all antibiotics recommended as relevant by regulatory agencies were short-listed and evaluated for their in silico and in vitro probiotic potentials. The four selected strains were assigned to the Bacillus velenzensis (MV4 and MV11), B. subtilis (MV24) and Priestia megaterium (formerly Bacillus megaterium) (MV30) species. A genomic analysis indicated that MV4, MV11 and MV24 contained a homolog of the gene coding for the fibrinolytic enzyme nattokinase while only MV30 encoded for a glutamic acid decarboxylase essential to synthesize the neurotransmitter GABA. All four strains showed a strong antioxidant activity, formed biofilm and produced/secreted quorum sensing peptides able to induce a cytoprotective stress response in a model of human intestinal (HT29) cells. Altogether, genomic and physiological data indicate that the analyzed strains do not pose safety concerns and have in vitro probiotic potentials.

Cashew (Anacardium occidentale) processing generates a by-product (CB) with potential for health benefits and could be a favorable ingredient to be added to a probiotic food matrix. This study aimed to assess the functional attributes of CB in fermented milk with a probiotic and a starter culture using in vitro- gastrointestinal conditions. Using a standardized fermentability assay, two strains more adapted to the dehydrated CB were chosen and applied in: Test Formulation (TF, with CB) and Control Formulation (CF, without CB). TF and CF were fermented by the probiotic strain Lacticaseibacillus paracasei subsp. paracasei F19® and the starter Streptococcus thermophilus ST-M6®. During a 28-day refrigerated (6-8 °C) shelf-life study, both strains used in CF and TF were resistant and maintained a population above 8.0 log CFU/mL in the fermented milk. TF had significant increase in total phenolic compounds and greater antioxidant potential during shelf life, along with improved survival of F19® after in vitro-simulated gastrointestinal conditions. The cashew by-product revealed to be promising as a substrate for the strains studied and the presence of CB increased the content of phenolics and antioxidant activity during storage, as well as the gastric resistance to the probiotic., leading to a promising healthy product.

regulating translational speed and accuracy. Threonylcarbamoyl adenosine (t6A37) and 5-methoxycarbonylmethyl-thiouridine (mcm5s2U34) are critical ASL modifications that have been linked to several human diseases. The model yeast Saccharomyces cerevisiae is viable despite the absence of both modifications, growth is however greatly impaired. The major observed consequence is a subsequent increase in protein aggregates and aberrant morphology. Proteomic analysis of the t6A-deficient strain revealed a global mistranslation leading to protein aggregation without regard to physicochemical properties or t6A-dependent or biased codon usage in parent genes. However, loss of sua5 led to increased expression of soluble proteins for mitochondrial function, protein quality processing/trafficking, oxidative stress response, and energy homeostasis. These results point to a global function for t6A in protein homeostasis very similar to mcm5/s2U modifications.

Abstract: Yeast – especially Saccharomyces cerevisiae – have long been a preferred workhorse for the production of numerous recombinant proteins and other metabolites. S. cerevisiae is a noteworthy aroma compound producer, and has also been exploited to produce foreign bioflavour compounds. In the past few years, important strides have been made in unlocking the key elements in the biochemical pathways involved in the production of many aroma compounds. The expression of these biochemical pathways in yeast often involves the manipulation of the host strain to direct the flux towards certain precursors needed for the production of the given aroma compound. This review highlights recent advances in the bioengineering of yeast – including S. cerevisiae – to produce aroma compounds and bioflavours. To capitalise on recent advances in synthetic yeast genomics, this review presents yeast as a significant producer of bioflavours in a fresh context and proposes new directions for combining engineering and biology principles to improve the yield of targeted aroma compounds.

The development of multidrug resistance by pathogenic bacteria and yeast is a significant medical problem that needs to be addressed. One possible answer could be the combined use of antibiotics and silver nanoparticles, which have different mechanisms of antimicrobial action. Nanoparticles can be combined with antifungal agents in the same way. Biogenic silver nanoparticles synthesized using environmentally friendly technology using extracts of biologically active plants are an effective nanomaterial that needs to be comprehensively investigated for implementation into medical practice. In this study, the synergistic effects arising from their combined use with antibiotics and fungicides against various bacteria and yeasts, included in the ESKAPE list, were studied. Effective concentrations of substances were established, recommendations for the studied pathogenic species were presented and the effect of destruction of the bacterial membrane was illustrated. The most significant synergistic effect was manifested in pathogenic candida and brewer's yeast.

The administration of Bacilli to dairy cows exerts a beneficial effect on dry matter intake, lactation performances and milk composition. In this work, two isogenic strains, only differing in the se-cretion level of cellulolytic and xylanolytic enzymes, were compared to establish whether B. subtilis cellulase and xylanase enzymes are involved in the positive effect exerted by these bacteria on animal nutrition. Conveniently, bacteria were grown in a newly designed growth medium based on an inexpensive waste. A multi-factorial study was conducted in which eight feed in-gredients were treated in vitro, using ruminal fluid from cannulated cows, with cultures of the two B. subtilis strains. Feed digestibility and gas production, a parameter linked to feed fermentability, were assessed. Fiber degradability was significantly higher in feeds treated with the en-zyme-overexpressing strain (10% and 5% higher than control treatment with B. subtilis-free media and the parent strain, respectively). This effect was accentuated for the most recalcitrant feeds, in which the benefits of cellulolytic and xylanolytic enzymes were maximized. Fermentability also benefited by the treatment with the enzyme-overexpressing strain, although the trend observed in gas production didn’t reach statistical difference between the two strains, and only minor changes in fermentation profiles were detected. Our results revealed that B. subtilis cellulases and xylanases effectively contribute to improving forage quality, supporting the use of Bacilli as fodder sup-plements to increase animal productivity.

In this experimental work, the microbiological, physicochemical, organoleptic and rheological characteristics of probiotic Bulgarian yoghurts produced by ultrafiltration of goat’s milk using different volume reduction ratio (2 and 3) and several probiotic starters (MZ2f, MZ2f + Bifidobacterium bifidum BB – 87, MZ2f + Lactobacillus acidophilus LAB – 8) were studied. It was established that the dry matter, protein, fat contents, the number of lactic acid bacteria, titratable acidity and the dynamic viscosity increased and the pH decreased with the rise of volume reduction ratio during ultrafiltration. All yoghurts exhibited Bingham plastic flow behavior. We recommend to use a volume reduction ratio of 3 and MZ2f + Bifidobacterium bifidum BB – 87 to produce probiotic Bulgarian yoghurts with the highest dry matter, protein, fat contents; counts of viable lactic acid cells, viscosity and the best sensory characteristics in the range of this experiment.

The [PSI⁺] nonsense-suppressor determinant of Saccharomyces cerevisiae is related to formation of heritable amyloids of the Sup35 (eRF3) translation termination factor. [PSI⁺] amyloids have variants in amyloid structure and in the strength of suppressor phenotype. Appearance of [PSI⁺], its propagation and manifestation depend primarily on chaperones. Besides chaperones, the Upf1/2/3, Siw14 and Arg82 proteins restrict [PSI⁺] formation, while Sla2 can prevent the [PSI⁺] toxicity. Here, we identify two more non-chaperone proteins involved in [PSI⁺] detoxification. We show that simultaneous lack of the Pub1 and Upf1 proteins causes lethality of [PSI⁺] cells with a strong, but not with weak suppressor phenotype. This lethality results from excessive depletion of the Sup45 (eRF1) termination factor due to its sequestration into Sup35 polymers. We also show that Pub1 acts to restrict excessive Sup35 prion polymerization, while Upf1 interferes with Sup45 binding to Sup35 polymers. These data allow considering the Pub1 and Upf1 proteins as a novel [PSI⁺] detoxification system.

Probiotics in cosmetics is a new and burgeoning area of research attracting attention from cosmetic manufacturers and customers looking for natural or additional alternatives in their skincare regimens. Probiotics not only promote beneficial microbiomes to improve skin health but can also modulate the visual, topographical, and mechanical attributes to improve skin quality. This study was conducted in healthy adults to determine the effect of a topical serum formulation containing the live probiotic Micrococcus luteus Q24 on several key cosmetic skin quality parameters and its beneficial impact on the modulation of the skin microbiome. Quantitative changes in various skin parameters were measured using a skin analyzer device and skin swabs were analysed using whole genome sequencing to monitor microbial diversity and levels of probiotic colonisation. Compared to the baseline, a significant reduction in the pore, spots, wrinkles, and impurities scores; and an increase of 101% in the hydration scores were observed. Additionally, 45-80% of participants showed a decrease in pores, spots, wrinkles, and impurities and 90% of participants showed an increase in hydration after 25 days of probiotic serum application. The WGS analysis of the skin swab samples showed a significant increase in the relative abundance of M. luteus Q24. The results from this study indicated that topical application of serum-containing probiotic M. luteus Q24 offers the benefit of improving skin health quality.

The microbiota in the oral cavity has a strict connection to its host. Its unbalance may determine oral diseases and can also have an impact on the systemic health. Probiotic strains may help in the restoration of a balanced condition. For this purpose, we screened the antibacterial and antiadhesive activities of many viable probiotic strains (Lactobacillus acidophilus PBS066, Lactobacillus crispatus LCR030, Lactobacillus gasseri LG050, Lactiplantibacillus plantarum PBS067, Limosilactobacillus reuteri PBS072, Lacticaseibacillus rhamnosus LRH020, Bifidobacterium animalis subsp. lactis BL050, Lacticaseibacillus paracasei LPC 1101, L. paracasei LPC 1082, and L. paracasei LPC 1114) against two main oral pathogens, Streptococcus mutans and Aggregatibacter actinomycetemcomitans, involved in dental caries and periodontal disease development and progression. Our results, obtained through agar overlay assay, plate count method, and scanning electron microscope imaging, indicate that the tested probiotics prevent the growth and adhesion of oral pathogens in a strain-specific manner, improving oral health. L. plantarum PBS067, L. rhamnosus LRH020, L. paracasei LPC 1101, L. paracasei LPC 1082, and L. paracasei LPC 1114 performed in a significant way against both pathogens in all the tests used. They can be considered as an effective adjuvant to guarantee oral and systemic well-being.

Plant diseases are among the major factors affecting plant productivity. Biological control of plant diseases is preferred over chemical control as it is environment-friendly, cost-effective, and sustainable. Among many microbes capable of providing biological control of plant diseases, probiotic Bacillus species are most promising as they can survive in adverse conditions, provide plants with a wide range of benefits including protection from phytopathogens. Wheat blast caused by Magnaporthe oryzae Triticum pathotype (MoT) has emerged as a potential threat to global wheat production. Due to unreliability of fungicides and limited cultivar resistance, we aimed to screen and identify potential antagonist bacteria collected from internal tissues of rice and wheat seeds to determine their in vitro and in vivo inhibitory effects against MoT. Dual culture and seedling assays were performed to evaluate the efficacy of probiotic bacteria. Out of 170 bacterial isolates, three bacteria (BTS-3, BTS-4, and BTLK6A) were screened as potential antagonists against MoT in vitro. Artificial inoculation at the seedling stage showed that the isolates BTS-4, BTS–3, and BTLK6A reduced 89, 88, and 85% of wheat blast disease severity, respectively, compared to mock-inoculated control. The bacterial isolates were identified as Bacillus subtilis (BTS-3) and B. velezensis (BTS-4 and BTLK6A) through genome phylogeny. The whole genome sequence of these three bacterial strains decoded a number of orthologs to intrinsic genes of antimicrobial peptides, antioxidant defense enzymes, cell wall degrading enzymes, compounds involved in the induction of systemic resistance (ISR) in host plants, and volatile compounds to make them promising biologicals to control MoT in wheat. Combined data of in vitro and in vivo along with genome analysis suggest that Bacillus spp. suppress the destructive wheat blast disease likely through antibiosis and ISR in the host plants. Further field evaluation and characterization of antimicrobial compounds are needed for a better understanding of the mode of action and practical recommendation of these bacteria for wheat blast control in the farmers’ fields.

The aim was to evaluate the influence of model (alcohol, sugar, salt, protein and acid) and real foods and beverages on the viability of probiotics during incubation and artificial digestion. Viability of monocultures Lactobacillus acidophilus CCM4833 and Bifidobacterium breve CCM7825T and commercial mixture of 9 probiotic bacterial strains were tested by cultivation assay and flow cytometry. In model foods, the best viability was determined in the presence of 0.2 g/L glucose, 10% albumin and 10% ethanol. As the most suitable real food for probiotic survival complex protein and carbohydrate substrates were found, such as beef broth, potato salad with pork, chicken with rice, chocolate spread, porridge and yoghurt. The best liquid was milk and meat broth, followed by coca-cola, beer and coffee. Viability of probiotics was higher when consumed with meals than with beverages only. Addition of prebiotics increased the viability of probiotics especially in presence of instant and fast foods. Generally, the highest viability of probiotics during artificial digestion was observed in mixed culture in presence of protein, sugar and fat or their combination. The increase of cell viability observed in such foods during model digestion may further contribute to the positive effect of probiotics on human health.

Improving antigen presentation is crucial for the success of immunization strategies. Yeasts are classically used as biofactories to produce recombinant proteins and are efficient vehicles for the delivery of vaccine antigens, besides present adjuvants properties. Despite the absence of epidemic outbreaks, several vaccine approaches continue to be developed for Zika virus infection. These prophylactic strategies are fundamental given the severity of clinical manifestations, mainly due to viral neurotropism. The present study aimed to evaluate in vivo the immune response induced by P. pastoris recombinant strains displaying epitopes of the Envelope (ENV) and NS1 ZIKV proteins. Intramuscular immunization with heat-attenuated yeast enhanced the secretion of IL-6, TNF-α, and IFN-γ, besides activation of CD4+ and CD8+ T cells, in BALB/c mice. P. pastoris displaying ENV epitopes induced a more robust immune response, increasing immunoglobulin production, especially IgG isotypes. Both proposed vaccines showed the potential to induce immune responses without adverse effects, confirming the safety of administering P. pastoris as a vaccine vehicle. Here we demonstrated, for the first time, the evaluation of a vaccine against ZIKV based on a multiepitope construct, using yeast as a vehicle, reinforcing the applicability of P. pastoris as a whole yeast cell vaccine.

Understanding the molecular basis of cancer initiation and progression is critical to develop effective treatment strategies. Recently, mutations in genes encoding histone proteins have been identified that drive oncogenesis, converting these essential proteins into “oncohistones”. Understanding how oncohistone mutations, which are commonly missense mutations, subvert the normal function of histones to drive oncogenesis requires defining the functional consequences of such changes. Histones genes are present in multiple copies in the human genome with 15 genes encoding histone H3 isoforms, the histone for which the majority of oncohistone variants have been analyzed thus far. With so many histone genes, engineering human cell lines that assess changes induced by sole expression of the oncohistone variant is technically challenging. In contrast to humans, budding and fission yeast contain only two or three histone H3 genes, respectively. Furthermore, yeast histones share ~90% sequence identity with human H3 protein. The genetic simplicity and evolutionary conservation make yeast an excellent model for characterizing oncohistones. The power of genetic approaches can also be exploited in yeast models to define cellular signaling pathways that could serve as actionable therapeutic targets. In this review, we focus on the value of yeast models to serve as a discovery tool that can inform subsequent translational studies in humans.

This study investigated a novel probiotic-enriched ice cream containing fermented white kidney bean homogenate to explore its potential health benefits. We assessed the viability of various probiotic strains during ice cream production and storage, focusing on their potential to reach the gut, and evaluated the overall antioxidant activity using DPPH, ABTS, FRAP, and TPC assays. The incorporation of fermented white bean homogenate significantly increased antioxidant capacity compared to the control group. Notably, strains like L. rhamnosus GG and L. plantarum 299v demonstrated the most pronounced effects on antioxidant activity, suggesting potential synergistic benefits between probiotics and bioactive compounds in fermented white beans. Although all probiotic strains experienced decreased viability during storage, certain strains, particularly L. plantarum 299v and L. casei DN-114001, showed promising survival rates even after 6 months. These results suggest the potential for developing probiotic ice cream containing viable bacteria capable of reaching the gut and contributing to a healthy gut microbiota. This could potentially improve digestion, reduce inflammation, and boost the immune system. Overall, this study highlights the potential of probiotic-enriched ice cream with fermented white kidney bean homogenate to combine the established benefits of probiotics for gut health with the enjoyment of consuming ice cream.

Concern for global health security and the environment due to the emergence of antibiotic-resistant bacteria and antibiotic residues in meat and other livestock products has led many countries to restrict the use of antibiotics in animal feed. This experiment was performed to assess the impact of dietary supplementation with a probiotic (Bacillus subtilis) and a postbiotic (Saccharomyces cerevisiae fermentation product) on growth performance, carcass traits, blood haemato-biochemical profile, gut microbiome, gut morphology, and immune response in broilers as an alternative to antimicrobials in poultry production system to minimize the effect on global health security. A total of 324-day-old Ven Cobb 400 broiler chicks were randomly divided into three dietary groups, each containing 12 replicated pens (n = 12), .and each replicate contained nine broiler chickens. The dietary groups consisted of 1) a basal diet without any growth promoting (T1), 2) the basal diet augmented with Bacillus subtilis at 200 g/MT feed (T2), and 3) the basal diet supplemented with Saccharomyces cerevisiae fermentation product at 1.25 kg/MT feed (T3). To calculate body weight gain, all birds and residual feed were weighed on a replicated basis over days 0, 7, 14, 21, 28, 35, and 42; mortality was recorded daily. Each replicate's mortality percentage was calculated and used to adjust BW, ADFI, and FCR calculations at the trial's end. At the end of the trial (42 d), two chickens from each replicate (24 chickens per group) were slaughtered for carcass traits, gut microbiome, and morphology measurements. Blood samples were collected for the haemato-biochemical profile on 35d and antibody titer on 28d and 35d.Feeding with SCFP (T3 group) significantly improved average daily feed intake (ADFI) and average daily gain (ADG) of chickens compared to the T1 (control) and T2 (probiotic) groups from 1 to 14 days of age. FCR was significantly improved (P<0.05) in SCFP-fed birds (T3) relative to the control (T1)over the entire experimental period (1-42 days). Carcass traits and blood haemato-biochemical parameters remained unaffected (P>0.05) by any diets. However, cholesterol levels and concentrations of corticosterone were significantly lower (P<0.05) in SCFP-fed birds (T3) compared to those of the probiotic (T2) and control (T1) groups. Total E. coli, Enterohaemorrhagic E. coli, ESBL-producing Enterobacteriaceae, and Salmonella counts were significantly higher in the T1group compared to T3 and T2 groups (P<0.05) and Salmonella counts were lower in T3 when compared to T2 (P<0.05). However, there was no significant difference in Lactobacillus count among treatment groups. PCR based characterization of ESBL- producing Enterobacteriaceae revealed maximum presence of blaCTX-M-Type (6/15, 40%), followed by blaSHV-Type (5/15, 33.3%) and blaTEM-Type (4/15, 26.6%). No variation in the occurrence pattern of ESBL/beta-lactamase associated genes in the studied ESBL- producing Enterobacteriaceae isolates was observed between the treatment groups. A significant increase in villi height and villi height to crypt depth ratio (VH: CD) (P<0.05) was observed in both T3 and T2 groups. Contrarily, villi widths did not vary across the dietary groups (P>0.05). The control group (T1) displayed a notable deeper in crypts when compared to the T3 and T2 groups (P<0.05). On day 28, the SCFP-fed birds (T3) and those fed the probiotics (T2) exhibited a significant increase (P<0.05) in antibody titers against Newcastle disease virus and Infectious bursal disease virus. Increase in antibody titre against IBDV in birds with the addition of SCFP in the diet was detected for the first time. However, there were no significant differences observed on day 35. The phagocytic activity of neutrophils and lymphocytes in broiler chickens did not show any noticeable variations among the different groups at day 35(P>0.05).It can be concluded that Saccharomyces cerevisiae fermentation product and Bacillus subtilis probiotic could be viable alternatives to antimicrobials in poultry production considering beneficial impacts in broilers fed antibiotic-free diet.

Almond (Prunus dulcis (Mill.) D.A.Webb) is one of the largest nut crops in the world. Recently, phenolic compounds, mostly stored in almond skin, have been associated with much of the health-promoting behavior associated with their intake. The almond skin enriched fraction obtained from cold-pressed oil residues of the endemic Moroccan Beldi ecotypes is particularly rich in chlorogenic acid. In this study, both almond skin extract (AE) and chlorogenic acid (CHL) supplements, similar to traditional positive control resveratrol, significantly increased the replicative life-span of yeast compared to the untreated group. Our results showed that AE and CHL significantly reduced the production of reactive oxygen and nitrogen species (ROS/RNS), most likely due to their ability to maintain mitochondrial function during aging, as indicated by the maintenance of normal mitochondrial membrane potential in treated groups. This may be associated with the observed activation of the anti-oxidative stress response in treated yeast, which results in activation at both gene expression and enzymatic activity levels for SOD2 and SIR2, the latter being an upstream inducer of SOD2 expression. Interestingly, the differential gene expression induction of mitochondrial SOD2 gene at the expense of the cytosolic SOD1 gene confirms the key role of mitochondrial function in this regulation. Furthermore, AE and CHL have contributed to the survival of yeast under UV-C-induced oxidative stress, by reducing the development of ROS / RNS, resulting in a significant reduction in cellular oxidative damage as evidenced by decreased membrane lipid peroxidation, protein carbonyl content and 8-oxo-guanine formation in DNA. Together, these results demonstrate the interest of AE and CHL as new regulators in the replicative life-span and control of the oxidative stress response of yeast.

Plant probiotic bacteria are being increasingly used to maximize productivity and quality of field crops. Pseudomonas chlororaphis O6 (PcO6) is a plant root colonizer with probiotic activities that produces metabolites such as phenazines functional in plant protection. This work reports responses of PcO6 to a non-ionic, triblock copolymer surfactant, Pluronic F68. This Pluronic exhibits membrane “healing” activity and improves cryopreservation recovery in eukaryotic cells. The product is FDA approved and is applied as an adjuvant in formulations used in agriculture, medicine, and biotechnology. Growth of PcO6 on Luria broth at 25°C was unhindered by 0.1 and 1.0 g/L F68, reduced at 10 g/L, with more inhibition at 100 g/L F68; micelle formation could account for inhibited growth at higher doses. Phenazine production was not changed by F68, whereas an F68 dose-dependent, surfactant-induced spread of bacterial colonization on 0.5% agar was observed. Exposure of cells to fluorescently labelled F68 resulted in intense fluorescence, stable to washing, showing a direct association of the Pluronic with the bacterium. However, neither protection nor harm was found for PcO6 cells undergoing repetitive freeze (-20 °C)/thaw cycles with 0.1% or 1% F68. These findings suggest F68 could be compatible for use in agricultural formulations with little effect on probiotics such as PcO6.

The aim of this work was to evaluate the influence of various food and beverages on the viability of probiotic bacteria during passing through artificial digestion. As a model food, solutions with various concentrations of alcohol, sugar, salt, protein and acid were prepared. Different types of real foods and beverages were used as well. Viability in presence of food matrices was tested on monocultures of Lactobacillus acidophilus CCM4833 and Bifidobacterium breve CCM7825T and on mixed commercial culture with 9 different strains of probiotic microorganisms( Lactobacillus, Bifidobacterium, Streptococcus). The concentration and viability of probiotic bacteria was tested by cultivation assay and flow cytometry. In model foods, the best growth of was determined in the presence of 10% albumin and 10% ethanol. Survival of the probiotics delivered in different food matrices through a simulated gastrointestinal tract was quantitatively different. As the best food environment for probiotics complex food matrices such as pasta with cream sauce, chocolate spread and homemade beef broth were selected, followed by mixed vegetables, potato salad, salted chips, fruits and yoghurt. Among beverages the best option was milk, followed by black tea, coffee and Coca Cola. Probiotic microorganisms are more viable when consumed with meals than with beverages only. In general, the highest viability of probiotic cells has been observed in presence of foods containing high concentration of sugar and fat or their suitable combination. The increase of cell viability observed in such foods during model digestion may further contribute to the positive effect of probiotics on human health.

For screening excellent lactic acid bacteria (LAB) strains to inhibit Escherichia (E.) coli (ETEC) K88, inhibitory activities of more than 1100 LAB strains isolated from different materials and kept in the lab were evaluated in this study. Nine strains with inhibition zone at least 22.00 mm (including that of hole puncher 10.00 mm) and good physiological and biochemical characteristics identified by 16S DNA gene sequencing and recA gene multiple detection, were assigned to Lactobacillus (L.) plantarum subsp. plantarum (5), L. fermentum (1), L. reuteri (1), W. cibaria (1) and E. faecalis (1), respectively. As investigated for their tolerance abilities and safety, only strain ZA3 possessed high hydrophobicity and auto-aggregation abilities, had high survival rate in low pH, bile salt environment and GI fluids, sensitive to ampicillin, resistant to norfloxacin and amikacin, without hemolytic activity and didn’t carry antibiotic resistance genes, exhibited broad spectrum activity against a wide range of microorganisms, and antibacterial substance may attribute to organic acids, especially lactic acid and acetic acid. The results indicated that the selected strain L. plantarum subsp. plantarum ZA3 could be considered a potential probiotic to inhibit ETEC K88 for further research.

Changes in consumer expectations have led to increasing demand for novel plant protection strategies, in order to reduce the application of chemical products, reduce the occurrence of new pests and the impact that all these actions generate in the environment. In recent years there have been numerous investigations related to biological control and the use of microorganisms as new control strategies. As part of integrated disease management, antagonistic microorganisms have been investigated lately and presented great interest. Such microorganisms can be applied in conventional and in organic farming as biological control agents (BCA). Many of these microorganisms are present in the microbial ecology generating interactive associations between surrounding microorganisms. For these reasons, it has become necessary to search new natural antimicrobial agents as alternatives to synthetic and chemical products. It has been discovered that there are microorganisms, particularly yeasts, that have antagonistic activity and different mechanisms of action, indicating that they could be interesting candidates for the development of BCA. Here, we evaluate the antagonist effect of four endophytic yeast, Cryptococcus antarcticus, Aureobasidium pullulans, Cryptococcus terrestris and Cryptococcus oeirensis over the growth of Botrytis cinerea, Monilinia laxa, Penicillium expansum and Geotrichum candidum in in vitro assays (inhibition zone diameter assay and confrontation assay).The results revealed that the four yeast strains evaluated showed antagonistic activity against the phytopathogens tested, suggesting that these yeasts produce compounds capable of inhibiting the growth of fungi and, depending on the assay, the evaluated antagonist-yeasts have differential biocontrolling-effect against the postharvest pathogens tested.

This research is aimed to investigate the effects of Lactoplantibacillus plantarum (L. plantarum) on growth performance, oxidation resistance, immunity, as well as cecal microbiota in broilers. This work classed three hundred and sixty 1-day-old broilers as three groups randomly, including control (CON, basal diet), antibiotic (ANT, 75 mg kg-1 chlortetracycline added into basal diet), or probiotic group (LP, 5×108 CFU kg-1 Lactoplantibacillus plantarum HJLP-1 contained within basal diet). Animal were then fed for 42 days, and each group consisted of 15 broilers for 8 replicates. Compared with CON, L. plantarum supplementation significantly improved the average weight daily gain (AWDG) (P < 0.05) while reducing feed-to-gain ratio over the entire supplemental period (P < 0.05). Birds fed L. plantarum had marked lower serum ammonia and xanthine oxidase levels (P < 0.05) than those in ANT and CON. Significant improvements (P < 0.05) of superoxide dismutase, catalase, serum IgM and IgY contents in broilers fed L. plantarum were also observed when compared those in CON and ANT groups. Both L. plantarum and antibiotics decreased the pro-inflammatory factor IL-1β level significantly (P < 0.05), while only L. plantarum promoted the anti-inflammatory factor IL-10 level dramatically in the serum (P < 0.05) compared with CON. The L. plantarum dramatically (P < 0.05) increased acetic acid and butyric acid concentrations in the cecal contents，when compared to those in CON and ANT. Among the differences revealed by 16S rRNA analysis, L. plantarum markedly improved the community richness of cecal microbiota. At the genus level, butyric acid-producing bacteria Ruminococcus and Lachnospiraceae were found in higher relative abundance in samples of L. plantarum treated birds. In conclusion, dietary L. plantarum supplementation promoted the growth and health of broilers, likely by inducing a shift in broiler gut microbiota toward short-chain fatty acid (SCFA)-producing bacteria. Therefore, L. plantarum has tremendous potential as an alternative for antibiotics in poultry breeding.

Cheese whey, a significant byproduct of the dairy industry, poses a substantial environmental challenge due to its organic pollutant load and large size demands for effective and affordable valorization methods. The abundance of lactose and other organic compounds in whey contributes to its elevated Biological Oxygen Demand (BOD) and Chemical Oxygen Demand (COD), further straining natural ecosystems. This study aimed to investigate the nutritional potential of whey and its conversion into value-added products: bioethanol and probiotic drinks. To achieve this, Bacillus megaterium and Bacillus subtilis strains, isolated from soil samples in Nepal, were co-cultured with Lactobacillus and Saccharomyces cerevisiae in the whey broth for fermentation. Ethanol distillation was carried out using a Rota-evaporator to maintain the viability of the fermenting organisms within the broth. Moreover, the probiotic criteria of the fermenting strains were extensively examined. Experimental observations revealed a remarkable concentration of 9.2 g/L of ethanol, resulting in an ethanol yield of 0.23 g/L. Extrapolating this rate, it is theoretically possible to produce an annual global bioethanol output of 5 million tons using whey as a sustainable and abundant resource. Furthermore, the study explores novel advancements in the distillation process, demonstrating the successful extraction of ethanol at room temperature. Additionally, the fermentation organisms employed in this research exhibit robust viability, surviving various in-vitro stress-tolerant tests, including exposure to bile salt, pH fluctuations, and gastric juice. These resilient strains also demonstrated long-term stability in fermented whey broth, making them promising candidates for the development of probiotic beverages. This dual approach not only enables the sustainable management of organic waste but also presents an opportunity to create value-added products with positive implications for both the environment and human health.

Enteropathogenic microorganisms like Escherichia coli cause severe intestinal problems by disrupting the gut homeostasis. The live microorganisms, when given in adequate quantities provide several beneficial effects to the host are known as probiotics. One of the pronounced benefits conferred by probiotic is to antagonize the growth of enteropathogens competing for adherence to the intestinal epithelium. Bifidobacterium is the major genera of human especially infant are intestinal microbiota. In current study, Bifidobacterium bifidum was isolated from the infant stools and probiotic potential was assessed using prescribed tolerance tests against low pH, gastric juices and bile salts. Anti-infectious activity of probiotic Bifidobacterium bifidum against enteropathogenic E. coli was checked both in vitro and in vivo using agar well diffusion assay and mice model respectively. Mice feces were evaluated for both Bifidobacterium bifidum and E. coli counts in all groups and analyzed statistically. In vitro results showed Bifidobacterium bifidum possess marked antibacterial activity against E. coli. There was significant decrease in enteropathogenic E. coli burden in the mice group fed with Bifidobacterium bifidum before and after challenge. In conclusion, the endogenous Bifidobacterium bifidum have excellent probiotic potential and can be used prophylactic and treatment option against enteropathogens.

The production of fuels and other industrial products from renewable sources has intensified the search for new substrates or for the expansion of the use of substrates already in use, as well as the search for microorganisms with different metabolic capacities. In the present work, we isolated and tested a yeast from the soil of sugarcane irrigated with vinasse, that is, with high mineral content and acidic pH. The strain of Meyerozyma caribbica URM 8365 was able to ferment glucose, but the use of xylose occurred when some oxygenation was provided. However, some fermentation of xylose to ethanol in oxygen limitation also occurs if glucose is present. This strain was able to produce ethanol from molasses substrate with 76% efficiency, showing its tolerance to possible inhibitors. High ethanol production efficiencies were also observed in acidic hydrolysates of each bagasse, sorghum, and cactus pear biomass. Mixtures of these substrates were tested and the best composition was found for the use of excess plant biomass in supplementation of primary substrates. It was also possible to verify the production of xylitol from xylose when the acetic acid concentration is reduced. Finally, the proposed metabolic model allowed calculating how much of the xylose carbon can be directed to the production of ethanol and/or xylitol in the presence of glucose. With this, it is possible to design an industrial plant that combines the production of ethanol and/or xylitol using combinations of primary substrates with hydrolysates of their biomass.

Many contaminant yeast strains able to survive inside fuel ethanol industrial vats show detrimental cell surface phenotypes, such as filamentation, invasive growth, flocculation, biofilm formation and excessive foam production. Previous studies have linked some of these phenotypes to the expression of FLO genes, and the presence of gene length polymorphisms causing the expansion of FLO gene size appears to result in stronger flocculation and biofilm formation phenotypes. We have performed here a molecular analysis of FLO1 and FLO11 gene polymorphisms present in contaminant strains of S. cerevisae from Brazilian fuel ethanol distilleries showing strong foaming phenotypes during fermentation. The size variability of these genes was correlated with cellular hydrophobicity, flocculation and highly foaming phenotypes in these yeast strains. Our results also show that deleting the major activator of FLO genes (the FLO8 gene) from the genome of a contaminant and highly foaming industrial strain avoids problematic foam formation, flocculation, invasive growth and biofilm production by the engineered (flo8∆::BleR / flo8Δ::kanMX) yeast strain. Thus, the characterization of highly foaming yeasts and the influence of FLO8 in this phenotype opens new perspectives for yeast strain engineering and optimization in the sugarcane fuel-ethanol industry.

Proper bipolar spindle assembly underlies accurate chromosome segregation. A cohort of microtubule-associated proteins orchestrates spindle microtubule formation in a spatiotemporally coordinated manner. Among them, the conserved XMAP215/TOG family of microtubule polymerase plays a central role in spindle assembly. In fission yeast, two XMAP215/TOG members, Alp14 and Dis1, share essential roles in cell viability; however how these two proteins functionally collaborate remains undetermined. Here we show the functional interplay and specification of Alp14 and Dis1. Creation of new mutant alleles of alp14, which display temperature sensitivity in the absence of Dis1, enabled us to conduct detailed analyses of a double mutant. We have found that simultaneous inactivation of Alp14 and Dis1 results in early mitotic arrest with very short, fragile spindles. Intriguingly, these cells often undergo spindle collapse, leading to a lethal “cut” phenotype. By implementing an artificial targetting system, we have shown that Alp14 and Dis1 are not functionally exchangeable and as such are not merely redundant paralogues. Intriguingly, while Alp14 promotes microtubule nucleation, Dis1 does not. Our results uncover that the intrinsic specification, not the spatial regulation, between Alp14 and Dis1 underlies the collaborative actions of these two XMAP215/TOG members in mitotic progression, spindle integrity and genome stability.

Background: Obesity and diabetes are two metabolic disorders linked by an inflammatory process named insulin resistance (IR). Various research on the role of gut microbiota in developing obesity and its associated disorders has led to the growing interest in probiotic supplementation. Considering the life-threatening complications of diabesity this mini-review explored the effects of probiotic supplementation on IR in obesity-associated diabetes. Method: This review is based on recent articles from 2005-2020, studying the role of probiotic supplementation on glucose and insulin parameters in healthy and diabetic mouse models. Result: Probiotic supplementation altered the gut microbiota composition, increased short-chain fatty acid production, and decreased pro-inflammatory cytokines. Additionally, they decreased intestinal permeability, circulating lipopolysaccharide, and metabolic endotoxemia hence improved insulin sensitivity and reduced obesity. Although multi-strain probiotic supplementation showed greater benefits than single strain interventions, variations in the concentration of probiotics used and the duration of treatment also influenced the results. Conclusion: Probiotic supplementation could manipulate the gut microbiota by reducing intestinal permeability, inflammation and ameliorate IR and obesity-associated diabetes in animal models which requires further long-term clinical studies in humans.

Cytokinesis, as the last stage of the cell division cycle, is a tightly controlled process amongst all eukaryotes, and defective division can lead to severe cellular consequences, and is implicated in serious human diseases and conditions such as cancer. Both mammalian cells and the fission yeast Schizosaccharomyces pombe use binary fission to divide into two equal sized daughter cells. Similar to mammalian cells, in S. pombe such cytokinetic division is driven by the assembly of an actin-myosin contractile ring (ACR) at the cell equator between the two cell tips. The ACR is composed of a complex network of membrane scaffold proteins, actin filaments, myosin motors, and other cytokinesis regulators. Contraction of the ACR leads to the formation of a cleavage furrow which is severed by the endosomal sorting complex required for transport (ESCRT) proteins leading to the final cell separation during the last stage of cytokinesis, abscission. This review describes recent findings defining the three phases to establish cytokinesis in S. pombe: ACR assembly, ACR constriction, and its coordination with septation. Collectively, we provide an overview of current understanding of the mechanisms regulating the ACR-mediated cytokinesis in S. pombe with a potential role of ESCRT proteins in this process.

This study was focused on the development of oil-producing microorganisms to generate lipids. The yeast Rhodotorula mucilaginosa (R. mucilaginosa) was selected for liquid-state cultivation, and the conditions for growth of the yeast cells were assessed. Additionally, the relationships between different nutrient elements and the growth of R. mucilaginosa were explored. The lipid accumulation of R. mucilaginosa is increased under nitrogen-restricted conditions. As the concentration of the carbon source increases, the accumulation of lipids is increased. However, if the carbon source concentration is further increased, the growth of yeast is inhibited. From a large-scale liquid fermentation culture with a fixed inoculation amount of 5%, and from a batch of culture experiments, it was determined that a suitable oil-producing culture was obtained on the 6th day, and the optimum conditions involved a carbon source concentration of 60 g/L, a nitrogen source concentration of 0.5 g/L, and a KH2PO4 concentration of 7.0 g/L. After utilizing different carbon sources in this study, it was found that glucose was the carbon source most conducive to the accumulation of R. mucilaginosa lipids. In addition, the extraction method and solvent for the extraction of R. mucilaginosa lipids were chosen. The acid-heat method using the green organic solvent ethyl acetate exhibited the best performance for extraction of yeast lipids under environmentally friendly and safe conditions. The analysis of lipids showed that the fatty acids obtained primarily contained C16:0, C18:1 and C18:2, and especially C18:1 (41%) lipids, indicating that R. mucilaginosa lipids are a good bio-oil source for the production of biodiesel.

Marine sediment sample was collected and the total aerobic population was counted and probiotic Bacillus cereus was isolated. Then it was mixed with formulated feed by spraying, while control diet was purchased from a local aquarium. 30 days of feeding experiment was conducted. Carassius auratus fishes were randomly divided into two experimental groups in rectangular tanks. The first group served as a control in which fishes were fed with commercial feed. The second group of fish were fed with probiotic incorporated feed. After 40 days, their survival and growth performance were detected. Bacillus cereus incorporated feed significantly yielded higher survival rate of the fish compared to control. Food conversion ratio (FCR) was significantly lower than the control group. The main aim of this work is to investigate the effect of a marine probiotic bacteria, Bacillus cereus on the survival and growth performance of gold fish Carassius auratus.

Red fruits such as grapes and blueberries are known for their high content of bioactive compounds and antioxidants. In Mediterranean winemaking, traditional sun drying can be replaced by controlled airflow chamber drying which provides better quality, higher phenolic content, and increased antioxidants. This study aimed to increase the sugar content and phenolic compounds of the must by drying the fruits to 50% of their original moisture content. Two musts were prepared by combining fresh grapes and dried blueberries (M1), and the other using dried grapes and fresh blueberries (M2), followed by fermentation at 25 ºC with M05 Mead and X5 yeast strains. Must M2 showed the highest level of phenolic compounds, A520, total anthocyanins and antioxidant activity. During fermentation, the anthocyanin content increased mainly in the dried blueberry macerates, which raised between 4 to 5.5-fold. More bioactive compounds were extracted from wines produced by yeast inoculation despite shorter maceration times. Sensory analysis demonstrated consumer acceptance of the wines in terms of color, flavor and aroma. In conclusion, the use of red grapes in the production of blueberry red wine proved to be effective, providing sugars and higher must yields, while the dried fruits improved the fermentable sugar content and the levels of bioactive compounds, increasing the antioxidant capacity of the resulting red fruit wines.

The aim of this study was to investigate succinic acid production from lactose concentrate, a by-product of cheese-making, using Actinobacillus succinogenes and Basfia succiniciproducens. Although the capability of these strains to metabolize different sugars is already known, their application in the conversion of lactose bears high potential for optimization. With regard to B. succiniciproducens this approach is completely novel. In particular the influence of the mediums sugar concentration and its supplementation with yeast extract to prevent a lack of proteins and vitamins were examined. Lactose based media containing sugar concentrations between 20 and 65 g L-1 and 5 g L-1 yeast extract were fermented, whereby both strains showed comparable performances. The best results in succinic acid yield and acid concentration, 0.57 g g-1 initial sugar and 23 g L1, were achieved at an initial sugar concentration of 43 g L-1. The necessity of yeast extract was demonstrated using the sugar optimized medium without supplementation. As a result, yield and concentration of succinic acid dropped to 0.34 g g-1 and 13 g L-1, the sugar consumption decreased from more than 99 to less than 55 %. Therefore the supplementation amount of 5 g L-1 yeast extract can be regarded as well-balanced.

Oxalate, a compound produced by many edible plants and as a terminal metabolite in the liver of mammals, is a toxin that has a detrimental role to human health. Humans and other mam-mals do produce the enzymatic machinery to degrade oxalate. However, numerous oxa-late-degrading bacteria reside in the mammalian gut and thus provide an important function for hosts. The current review focuses on the environmental factors that influence the efficacy of pro-biotic oxalate-degrading bacteria, relative to oxalate metabolism. We describe the mechanism of oxalate catabolism and its consumption by obligate and facultative anaerobic oxalate-degrading bacteria, in both in vitro and in vivo environments. We also explore the environmental variables that impact oxalate degradation. Studies on single species degrade oxalate have not shown a strong impact on oxalate metabolism especially in high oxalate conditions such as consumption of foods high in oxalate (such as coffee and chocolate for humans or halogeton in animal feed). Considering effective variables which enhance oxalate degradation could be used in application of effective probiotic as a therapeutic tool in individuals with hyperoxaluria. This study indicates probiotics can be considered a good source of naturally occurring oxalate degrading agent in human colon

Magnesium oxide is typically white and can be colorized with transition metal insertion by doping or by adsorption. We present the preparation of green colored hydroxide by the adsorption of nickel ions by porous MgO. MgO was prepared by the colloidal starch suspension method and used to remove nickel ions from the aqueous solution. The oxides and hydroxides, before and after adsorption, were characterized by X-ray diffraction technical, and show that a phase change occurs, a transition from periclase (MgO) to brucite (Mg(OH)2) due to the interaction with nickel salts (acetate, chloride, and nitrate). The FTIR spectrum corroborates with the DRX data on the formation of brucite like crystal lattices. The new samples present a green color, indicative of the incor-po-ration of the Ni2+ ions. Mixed oxides in the form of brucite were used in microbiological tests and showed action against fungi and bacteria, with bactericidal, bacteriostatic, or fungistatic function. Green brucite (NixMg1-x(OH)2) can be used as an inorganic pigment in white or colorless paints with a special biocidal property.

The study investigated the effect of a multi-strain probiotic compound containing Bacillus coagulans, Enterococcus faecalis, Clostridium butyricum and Bacillus mesentericus as in-feed zinc oxide (ZnO) alterna-tive on growth performance, diarrhea incidence, antioxidant status, lipid profile, stress and immunity in weaned piglets. A total of 72 piglets were randomly divided into 3 groups with four replicates of six piglets each. The details of the groups were as follows; (i) weaned control group (WC) received basal diet, (ii) weaned probiotic group (WB) received basal diet and probiotics and (iii) positive control (PC) group received basal diet with 2500 mg/kg ZnO. The experiment lasted for 28 days. Probiotic supple-mentation improved growth performance and reduced diarrhea rate. Probiotics supplementation im-proved lipid profile; significantly lower levels of total cholesterol and low-density lipoprotein cholesterol and higher level of high-density lipoprotein cholesterol in WB group as compared to those of the control group (WC) were recorded. Probiotic supplementation stimulated antioxidant defense system by in-creasing total antioxidant capacity and decreasing lipid peroxidation. Probiotic supplementation down-regulated the stress biomarkers like serum cortisol and serum heat shock proteins. WB group showed higher serum levels of IgG and IgM throughout the study period and higher IgA at day 28 as compared to WC. These data suggest that supplementation of the probiotic minimizes the weaning stress, thereby improves the growth performance, lipid profile, antioxidant status and systemic as well as mucosal immunity. Therefore, the multi-strain probiotic compound may be used to replace ZnO in weaned piglets.

Saccharomyces cerevisiae cells are killed by zymocin, a tRNase ribotoxin complex from Kluyveromyces lactis, which cleaves anticodons and inhibits protein synthesis. Zymocin’s action requires specific chemical modification of uridine bases in the anticodon wobble position (U34) by the Elongator complex (Elp1-Elp6). Hence, loss of anticodon modification in mutants lacking Elongator or related KTI (K. lactis Toxin Insensitive) genes protects against tRNA cleavage and confers resistance to the toxin. Here, we show that zymocin can be used as a tool to genetically analyse KTI12, a gene previously shown to code for an Elongator partner protein. From a kti12 mutant pool of zymocin survivors, we identify motifs in Kti12 that are functionally directly coupled to Elongator activity. In addition, shared requirement of U34 modifications for nonsense and missense tRNA suppression (SUP4; SOE1) strongly suggests that Kti12 and Elongator cooperate to assure proper tRNA functioning. We show that the Kti12 motifs are conserved in plant ortholog DRL1/ELO4 from Arabidopsis thaliana and seem to be involved in binding of cofactors (e.g. nucleotides, calmodulin). Elongator interaction defects triggered by mutations in these motifs correlate with phenotypes typical for loss of U34 modification. Thus, tRNA modification by Elongator appears to require physical contact with Kti12, and our preliminary data suggest that metabolic signals may affect proper communication between them.

“All diseases originate in the gut.” Hippocrates (400 BC) A healthy gut microbiome via the gut-brain-axis (GBA) elevates heart rate variability (HRV), a general measure of health and well-being. A dysbiotic gut microbiome, low in biodiversity and butyrate producers, alters tryptophan metabolism with release of proinflammatory cytokines, predominantly TNF-α, IL-6, and IL-1β. These also characterize chronic inflammation, oxidative stress, and a multitude of diseases, all exhibiting low HRV. Gut dysbiosis upregulates IFN-γ and with it IDO (indoleamine 2,3 dioxygenase). Tryptophan pivots from serotonin synthesis to that of IDO induced kynurenine, increasing the kynurenine to tryptophan ratio (KTR). An elevated KTR is positively linked to neurodegenerative and autoimmune diseases and negatively linked to HRV. Elevated IDO activity is not only enzymatic but also an intracellular signal transducer potentiated by TGF-β. This cytokine is the primary determinant of the TME. Also proposed is the gut-lung dysbiosis concept and consequent degradation of ACE2 (richest in lungs and gut). Leaky gut induced autoantibodies related to G-protein coupled receptors (GPCRs) in combination with increased Ang II further potentiate oxidative stress. Aldosterone and paroxysmal orthostatic tachycardia syndrome (POTS) paradoxes are highlighted in the context of GPCR and gut dysbiosis, and the role of Candida is explored. The triple play of a prebiotic (d-mannose), probiotic (bifidobacteria and lactobacilli), and postbiotic (butyrate) might improve intestinal barrier integrity, oppose entry of GPCR antigens, suppress the inflammatory cytokine triad, balance IFN-γ and TGF-β, suppress oxidative stress, depress KTR, elevate HRV, and extend lifespan and its quality.