Search Results (93)

This study is Part II of a five-year (2018–2022) field trial in western Portugal evaluating the effects of three microbial biofertilizers—Mycoshell^® (Glomus spp. + humic/fulvic acids), Kiplant iNmass^® (Azospirillum brasilense, Bacillus megaterium, Saccharomyces cerevisiae), and Kiplant All-Grip^® (Bacillus megaterium, Pseudomonas spp.)—applied at different dosages alongside two mineral fertilizer regimes, T100 (full dose) and T70 (70% of T100, alone or combined with biofertilizers), on the physiological performance of ‘Gala Redlum’ apple trees. Part I had shown that Myc4 (Mycoshell^®, 4 tablets/tree), iNM6, and iNM12 (Kiplant iNmass^®, 6 and L ha⁻¹, respectively) consistently enhanced fruit growth, yield, and selected quality traits. While Part I showed clear agronomic gains, Part II demonstrates that these improvements occurred without significant alterations in seasonal photosynthetic performance, canopy reflectance, or chlorophyll fluorescence parameters over five years, highlighting the contrast between observed yield improvements and physiological stability. Seasonal monitoring of physiological traits—including specific leaf area (SLA), chlorophyll content index (CCI), gas exchange (A_n, g_s, E, C_i), spectral indices (NDVI, OSAVI, SIPI, GM2), and chlorophyll fluorescence (OJIP). It is clear that physiological values remained largely stable across biofertilizer treatments and years. Importantly, this stability was maintained even under a 30% reduction in mineral fertilizer (T70), indicating that specific microbial biofertilizers can sustain physiological resilience under reduced nutrient inputs, thereby providing a physiological basis for the yield-enhancing effects observed and supporting their integration into fertilizer reduction strategies in Mediterranean orchards. Full article

Beer is a drink that has been a staple in human history, evolving from its beginning in antiquity to the present day. Nowadays, large breweries and other companies have set up laboratories focused on finding and developing new yeast strains for the brewing sector to meet consumers’ demand for new beer styles. Monascus spp. are ascomycota that have been known for hundreds of years. They are widely popular in Asian cuisine, especially in fermented foods. Studies show that Monascus spp. produce numerous food dyes and substances that positively influence human health. In the presented work, Monascus ruber was tested as a potential microorganism for the beer industry. Experiments included fermentation trials with Monascus ruber in four regimes: in aerobic condition, anaerobic condition, anaerobic condition with pH kept above 4.5, and in anaerobic condition with pH set to 4.5. As a reference, commercial Saccharomyces cerevisiae and Saccharomyces pastorianus were used. Fermentation parameters were evaluated by measurements of ethanol and extract level. The final product was tested for its colour in order to evaluate if monascus-derived pigments were present in the beverage. Moreover, a qualitative analysis of lovastatin and citrinin was performed in order to check if those monascus metabolites were present. Finally, small-scale consumer tests were performed in order to check the organoleptic properties of the obtained beverage. Results show that Monascus ruber is able to ferment beer wort in a similar manner as Saccharomyces strains, reaching a slightly lower degree of attenuation. Nevertheless, a longer lag phase was observed in monascus trials, except for the trial with preset pH at 4.5. The most visible change in the product was a reddish colour that appeared in the sample in aerobic conditions. The qualitative analysis showed that lovastatin and citrinin were present in the tested samples. Consumer tests show that experimental beer has a different taste than Saccharomyces-fermented products. Although the presented results are preliminary, they could be a good starting point for further research on monascus-based beverages. Full article

Due to the neuroprotective and antioxidant properties, phenylethanoid glycosides (PhGs) are valuable plant-derived compounds. Traditional extraction methods are constrained by low yields and limited resources, prompting the integration of synthetic biology and enzyme engineering technologies for sustainable production. This review summarizes the advances in the microbial synthesis of PhGs, emphasizing the elucidation of biosynthetic pathways, enzyme engineering modifications of glycosyltransferases and acyltransferases, and strategies for optimizing microbial cell factories in Escherichia coli and Saccharomyces cerevisiae. Significant advancements encompass the efficient synthesis of verbascoside and echinacoside in S. cerevisiae, as well as the comprehensive elucidation of the echinacoside biosynthetic pathway in Cistanche spp., including the identification of key steps catalyzed by a rhamnosyltransferase, a CYP450 hydroxylase, and a terminal glucosyltransferase that enable pathway reconstruction in S. cerevisiae. We conduct a systematic analysis of methods to address the biosynthetic bottlenecks via protein engineering, including rational design and directed evolution, as well as the metabolic engineering strategies such as precursor enhancement and cofactor recycling. Additionally, we investigate the synthesis of non-natural PhG analogues and the prospective integration with AI-assisted design, emphasizing the significant potential of microbial systems in overcoming the supply challenges for medicine-food homologous ingredients. Full article

Biofertilizers are sustainable alternatives to mineral fertilizers in perennial crops, reducing the need for mineral inputs. This five-year field study evaluated three biofertilizers—Mycoshell^® (Glomus spp. + humic/fulvic acids), Kiplant iNmass^® (Azospirillum brasilense, Bacillus megaterium, and Saccharomyces cerevisiae), and Kiplant All-Grip^® (Bacillus megaterium and Pseudomonas spp.)—at different dosages alongside two mineral fertilizer regimes, T100 (full recommended dose) and T70 (70% of T100, alone or combined with biofertilizers), in an apple orchard under Mediterranean conditions. Biofertilizers maintained or increased soil nutrient availability by 5–15% and leaf N, P, K, Mg, and Zn concentrations by 5–12% compared with T100. Trees under biofertilizers, particularly Myc2 and Myc4, exhibited greater shoot growth (up to 30.4 m/year), trunk cross-sectional area (TCSA: 11.9 cm² in 2022), and canopy volume (2.21 m³), representing 10–20% increases. Selected biofertilizer treatments produced 6–7.5 kg/tree, 130–145 g average fruit weight, 66–74 mm diameter, 13.9–18.7 °Brix, and 13–18% dry matter, maintaining >90% of yield and fruit size relative to T100, with more balanced medium- and large-sized fruit distribution. Principal Component Analysis explained 66–72% of soil and leaf nutrient variance, confirming their multivariate impact. Overall, biofertilizers applied at recommended doses and timings can partially replace mineral fertilizers, sustaining productivity and quality, enhancing nutrient availability, and supporting long-term orchard sustainability. While climate remains the main driver of annual production, these findings provide evidence for integrating biofertilizers into environmentally friendly fertilization strategies. Full article

Increasing interest in functional nutrition has driven the incorporation of probiotics into pet food formulations to enhance digestive health, immune response, and overall well-being in companion animals. This systematic review examines scientific publications, patents, and market developments related to probiotic-enriched pet food from 2014 to 2024. We evaluate major probiotic taxa—including Lactobacillus spp., Bifidobacterium spp., Bacillus spp., and Saccharomyces cerevisiae—based on their resilience during processing, gastrointestinal survival, and documented health benefits. Delivery technologies such as microencapsulation, coating matrices, and post-processing supplementation are analyzed for their effectiveness in preserving probiotic viability within dry and wet food matrices. Patent landscape analysis highlights innovation trends in strain selection, formulation design, and processing methods. In vivo and in vitro studies demonstrate that probiotic supplementation modulates gut microbiota composition, improves fecal parameters, enhances immune markers, and promotes nutrient absorption in both canine and feline models. Market data reveal rapid expansion of commercial probiotic pet food products, yet scientific research remains limited compared to human nutrition. Overall, the findings indicate that while probiotics hold clear potential to improve gastrointestinal health and immunity in pets, evidence remains fragmented, particularly for cats and long-term outcomes. Bridging the gap between industrial innovation and controlled clinical validation will be essential for developing next-generation probiotic pet foods. Full article

Non-Saccharomyces yeasts are emerging as promising new probiotics with a beneficial effect equal to or greater than the reference probiotic yeast, Saccharomyces boulardii. Candida intermedia, a non-albicans species not considered a common human pathogen, previously demonstrated probiotic potential. In this work, our objective was to evaluate the immunomodulatory effects of C. intermedia ORQ001 in mice vaccinated with inactivated SARS-CoV-2, seeking further evidence of its probiotic activity. Murine macrophages were stimulated with C. intermedia, followed by mRNA transcription analysis via qPCR. Mice were supplemented with C. intermedia prior to SARS-CoV-2 vaccination. Antibody production was assessed by ELISA, and fecal microbiomes were analyzed using next-generation sequencing. C. intermedia significantly increased Il4 and Il13 expression while decreasing Stat3 in macrophages. Splenocytes from supplemented mice exhibited elevated transcription levels of Tnf, Ifng, Il4, Bcl6, and Stat3 after exposure to stimulatory molecules. These mice showed increased levels of anti-SARS-CoV-2 IgG and sIgA isotypes, along with higher abundances of Bacteroides spp. and Clostridium spp. in their gut microbiome. In conclusion, C. intermedia supplementation modulated the expression of key immune-related genes and enhanced humoral responses in mice. Furthermore, its influence on gastrointestinal microbiota suggests a synergistic effect on vaccine immunogenicity. These findings support the potential of C. intermedia as a novel probiotic candidate with immunomodulatory properties applicable to vaccine adjuvanticity. Full article

Although beer fermentation has traditionally been carried out with Saccharomyces, the boom in craft brewing has led to the use of non-conventional yeast species for beer production. This group also includes non-Saccharomyces starters, which are commonly used in winemaking and which have different technological characteristics compared to standard representatives of the Saccharomyces genus. One of the important characteristics of the non-Saccharomyces group is the richer enzyme profile, which leads to the production of beverages with different taste and aroma profiles. The aim of this study was to investigate sweet and hopped wort fermentation with seven strains of active dry non-conventional yeasts of Lachancea spp., Metschnikowia spp., Torulaspora spp. and a mixed culture of Saccharomyces cerevisiae and Torulaspora delbrueckii. One ale and one lager active dry yeast strain were used as control strains. The extract consumption, ethanol production, degree of fermentation, pH drop, as well as the yeast secondary metabolites formed by the yeast (higher alcohols, esters and aldehydes) in sweet and hopped wort were investigated. The results indicated that all of the studied types of non-conventional yeasts have serious potential for use in beer production in order to obtain new beer styles. For the purposes of this study, statistical methods, principle component analysis (PCA) and correlation analysis were used, thus establishing the difference in the fermentation kinetics of the growth in the studied species in sweet and hopped wort. It was found that hopping had a significant influence on the fermentation kinetics of some of the species, which was probably due to the inhibitory effect of the iso-alpha-acids of hops. Directions for future research with the studied yeast species in beer production are presented. Full article

This study was undertaken to identify foodborne bacteria and fungi from different parts of eggs depending on their storage duration, organoleptic properties, total viable count, and antibiotic resistance profile. Thirty-two samples were randomly collected from commercial layer farms in Mymensingh. Following the protocol of sample preparation, outer-surface and inner-content samples were streaked onto various selective media. Isolation and identification were carried out by observing Gram staining and biochemical properties. Molecular detection was confirmed through a PCR assay using specific primers for Salmonella spp., E. coli, Staphylococcus spp., and fungus (Simplicillium spp. and Saccharomyces spp.). To determine the antibiotic resistance profile, the disk diffusion method was followed against nine antibiotic disks. The isolation rate of E. coli, Salmonella spp., and Staphylococcus spp. was 53.13%, 40.63%, and 40.63%, respectively, in the outer eggshell and 15.63%, 25%, and 15.63%, respectively, in the inner content of the eggs. Regarding the fungus content (yeast and mold), 100% was obtained in the outer eggshell, whereas there was an absence of fungus in the inner content. It was observed that all the isolates of E. coli, Salmonella spp., and Staphylococcus spp. were highly sensitive to either Ciprofloxacin or Levofloxacin and extremely resistant to Amoxicillin or Azithromycin drug disks or both. The data also shows that storage duration had a proportional relationship with TVC and an inversely proportional relationship with organoleptic properties. This study indicates that eggs harbor multidrug-resistant foodborne bacteria, which might constitute a public health hazard if these antibiotic-resistant bacteria are transferred to humans. Full article

Winemaking involves a microbial ecosystem where yeast diversity, shaped by terroir and winemaking conditions, determines wine characteristics. Understanding the microbial diversity of vineyards and spontaneous fermentation is crucial for explaining a winery’s typical wine profile. Studying and inoculating indigenous strains make it possible to produce high quality wines, reflecting the production environment. This study analyzes the yeast species involved in 16 spontaneous fermentations (8 in 2022 and 8 in 2023) from grapes of four distinct vineyards under two sets of winemaking conditions. A total of 1100 yeast colonies were identified by MALDI-TOF and DNA sequencing techniques. Saccharomyces (S.) cerevisiae and Hanseniaspora uvarum were the most prevalent species, alongside significant populations of non-Saccharomyces yeasts such as Lachancea thermotolerans and Metchnikowia pulcherrima, which were the most abundant ones. Minor yeast species, including Aureobasidium pullulans, Starmerella bacillaris, Kazachstania servazzi, and other Hanseniaspora spp., were also detected. The results demonstrated that yeast diversity in spontaneous fermentations varied according to vineyard origin and winemaking conditions. Differences between the two vintages studied indicated that annual climatic conditions significantly influenced yeast diversity, especially among non-Saccharomyces species. This substantial diversity represents a valuable source of indigenous yeasts for preserving the typicity of a winery’s wines under controlled conditions. Full article

CRISPR technology, which is derived from the bacterial adaptive immune system, has transformed traditional genetic engineering techniques, made strain engineering significantly easier, and become a very versatile genome editing system that allows for precise, programmable modifications to a wide range of microbial genomes. The economies of fermentation-based manufacturing are changing because of its quick acceptance in both academic and industry labs. CRISPR processes have been used to modify industrially significant bacteria, including the lactic acid producers, Clostridium spp., Escherichia coli, and Corynebacterium glutamicum, in order to increase the yields of bioethanol, butanol, succinic acid, acetone, and polyhydroxyalkanoate precursors. CRISPR-mediated promoter engineering and single-step multiplex editing have improved inhibitor tolerance, raised ethanol titers, and allowed for the de novo synthesis of terpenoids, flavonoids, and recombinant vaccines in yeasts, especially Saccharomyces cerevisiae and emerging non-conventional species. While enzyme and biopharmaceutical manufacturing use CRISPR for quick strain optimization and glyco-engineering, food and beverage fermentations benefit from starter-culture customization for aroma, texture, and probiotic functionality. Off-target effects, cytotoxicity linked to Cas9, inefficient delivery in specific microorganisms, and regulatory ambiguities in commercial fermentation settings are some of the main challenges. This review provides an industry-specific summary of CRISPR–Cas9 applications in microbial fermentation and highlights technical developments, persisting challenges, and industrial advancements. Full article

Modified nano-clays, alone or combined with probiotics, may offer a novel and sustainable approach to improve ruminal fermentation and mitigate CH₄ emissions in high-fiber diets. This study evaluated the properties and effects of modified nano-bentonite (MNB), with or without yeast (Saccharomyces cerevisiae), compared to natural bentonite (NB) and monensin, using the in vitro gas production (GP) technique. The substrate used was a basal diet composed primarily of forage (Trifolium alexandrinum clover) in a 70:30 forage-to-concentrate ratio. The treatments were a control group receiving the basal diet without additives; a monensin-added diet containing 40 mg/kg of dry matter (DM); a yeast-added diet with Saccharomyces cerevisiae at 2 × 10⁸ CFU/g of DM; a NB clay-added diet at 5 g/kg of DM; and MNB diets added at two levels (0.5 g/kg of DM (MNB_Low) and 1 g/kg of DM (MNB_High)), with or without S. cerevisiae. MNB showed a smaller particle size and improved properties, such as higher conductivity, surface area, and cation exchange capacity, than NB. Sulfur and related functional groups were detected only in MNB. No differences were observed in total GP, while both the monensin diet and the MNB_High-with-yeast diet significantly reduced CH₄ emissions compared to the control (p < 0.05). The MNB_High-without-yeast combination significantly (p < 0.05) reduced hemicellulose degradation, as well as total protozoal counts, including Isotricha and Epidinium spp. (p < 0.05), compared to the control. Ammonia levels did not differ significantly among treatments, while NB and MNB_High diets tended to have (p = 0.063) the highest short-chain fatty acid (SCFA) concentrations. These findings suggest the potential modulatory effects of yeast and MNB on rumen fermentation dynamics and CH₄ mitigation. Full article

Sugarcane (Saccharum spp. L.), traditionally cultivated in tropical and subtropical regions, is being explored for its agronomic viability in Mediterranean climates. This study assessed the bio-agronomic performance of seven sugarcane varieties and two accessions grown in Sicily, to enhance the fermentation process to produce rum agricole, a spirit derived from fresh cane juice. Agronomic evaluations revealed significant varietal differences, with juice yields of 5850−14,312 L ha⁻¹ and sugar yields of 1.84–5.33 t ha⁻¹. Microbial control was achieved through the addition of lactic acid, which effectively suppressed undesirable bacterial growth and improved fermentation quality. Furthermore, the application of two selected Saccharomyces cerevisiae strains (MN113 and SPF21), isolated from high-sugar matrices such as manna and honey byproducts, affected the production of volatile compounds, particularly esters and higher alcohols. Sensory analysis confirmed a more complex aromatic profile in cane wines fermented with these selected yeasts, with overall acceptance scores reaching 7.5. Up to 29 aroma-active compounds were identified, including ethyl esters and higher alcohols. This research represents the first integrated approach combining lactic acid treatment and novel yeast strains for the fermentation of sugarcane juice in a Mediterranean context. The findings highlight the potential for high-quality rum agricole production in Sicily. Full article

Previous studies have shown the positive effects of non-conventional Metschnikowia spp. yeasts in mixed cultures with Saccharomyces cerevisiae on the properties of fruit wines. In this study, we investigated the effects of using conventional S. cerevisiae and non-conventional Metschnikowia pulcherrima yeasts as starter cultures in controlled mixed fermentations of honey wort. Other non-conventional yeasts were also tested for comparison, including Wickerhamomyces anomalus, Dekkera/Bretannomyces bruxellensis, and Wickerhamomyces anomalus. We evaluated the tolerance of the tested yeasts to high sugar content and analyzed the metabolic profiles of both monocultures and mixed systems. The M. pulcherrima strain showed the highest tolerance to 30% w/v glucose. The chemical complexity of fermented honey was improved using M. pulcherrima in co-starters with S. cerevisiae. The fermented honey samples were characterized by lower ethanol content, higher glycerol level, and rich volatilomes containing higher levels of both esters (ethyl acetate, 3-methylbutyl acetate, 2-methylpropyl acetate) and aliphatic alcohols (2-methylpropan-1-ol, 3-methylbutan-1-ol, and 2-methylbutan-1-ol). Similar characteristics were obtained using mixed populations of four strains: S. cerevisiae, M. pulcherrima, D. bruxellensis, and W. anomalus. Full article

This study employed shotgun metagenomics to investigate microbial dynamics, phage-bacteria interactions, and functional genes throughout a three-month apple vinegar fermentation process. A total of 5621 microbial species were identified, revealing three distinct phases: (i) Enterobacteria and non-Saccharomyces species dominated the initial substrate; (ii) S. cerevisiae and Leuconostoc pseudomesenteroides prevailed in the intermediate phase; and (iii) acetic acid bacteria (Acetobacter ghanesis and Gluconobacter spp.), alongside non-Saccharomyces species (Pichia kudriavzevii and Malassezia restricta), dominated the final stages. Bacteriophage analysis revealed the presence of phages targeting spoilage bacteria, such as Pseudomonas and Erwinia, suggesting a role in regulating microbial stability and enhancing fermentation control. Functional metagenomic analysis highlighted key pathways associated with microbial growth and metabolite production, including carbohydrate and amino acid metabolism, energy production, and glycan biosynthesis. Enzymes involved in stress adaptation and secondary metabolism, including oxidative phosphorylation and phenolic compound synthesis, demonstrated microbial resilience and their potential role in shaping the product’s sensory and functional properties. Moreover, Enterobacteriaceae species were associated with pectin degradation during the early stages, aiding substrate breakdown. These findings are crucial for microbial and phage management in fermentation technology, offering valuable insights for innovation in the vinegar industry. Full article

Insect-based (silkworm cocoons) and plant-based (cotton wool pads and gauzes) fiber substrates were used to support and ameliorate seed germination originating from trifoliate orange (Poncirus trifoliata) and pomegranate (Punica granatum) trees. Three different commercial formulations of beneficial microorganisms (Bacillus spp.-Azotobacter spp., Saccharomyces boulardii, and Saccharomyces cerevisiae) were administered to seeds in order to evaluate their contribution to germination and growth. The silkworm cocoons provided better germination rates for P. trifoliata seeds (83.33%) among the tested media without any microbial supplementation. These rates increased towards the absolute maximum (100%) when Bacillus spp.-Azotobacter spp., S. boulardii and S. cerevisiae were applied. Furthermore, inoculums of Bacillus spp.-Azotobacter spp. 2 mL and S. cerevisiae 3 g raised the pomegranate seed germination ability by 30–33.33% and 50–67.7%, respectively, on silkworm cocoon substrates when compared to plant-derived, cellulosic fiber substrates under the same biotic exposure. On increasing the size of applied microbial inoculums, seed germination moved from optimum to suboptimum for all germination media. Examination of multipartite pH compatibility (between seeds, microorganisms, and germination media) was beneficial and of functional value. In conclusion, the germination rates of both tree species can be raised using bacterial and yeast supplementation, including medical-grade S. boulardii, on environmentally friendly materials such as insect- and plant-based fiber substrates. Full article