Search Results (307)

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

Grapevine (Vitis vinifera) is a key crop in Mediterranean agriculture, now increasingly threatened by Xylella fastidiosa subsp. Fastidiosa (Xff), the causal agent of Pierce’s disease. This study investigated: (1) the diversity of culturable fungal endophytes in the xylem sap of naturally Xff-infected grapevines, and (2) the interaction between Xff and the pathogenic fungus Sclerotinia sclerotiorum identified in the sap. The xylem sap was collected from Cabernet Sauvignon vines in Mallorca, Spain, and fungal communities were characterized using culture-dependent methods. Both beneficial fungi (e.g., Aureobasidium pullulans, Rhodotorula mucilaginosa) and pathogenic species (e.g., S. sclerotiorum, Cladosporium sp., Alternaria alternata, and the Phoma complex) were isolated from both Xff-positive and Xff-negative plants, indicating similar community profiles. Although limited by small sample size, these findings offer preliminary evidence of complex ecological interactions between Xff and the xylem-associated mycobiota, with potential implications for grapevine health and disease development under varying environmental and management conditions. Further experiments under controlled conditions revealed that grapevines co-inoculated with Xff and S. sclerotiorum showed increased disease severity, suggesting a synergistic interaction. These preliminary results highlight the complex interplay between Xff and the fungal endophytic microbiome, which may modulate grapevine susceptibility depending on environmental and management conditions. Full article

Global concerns about environmental pollution, poor waste management, and the rise in antimicrobial resistance due to uncontrolled antibiotic use have driven researchers to seek alternative, multifaceted solutions. Plants, animals, microorganisms, and their processing wastes serve as valuable sources of natural biopolymers and bioactive compounds. Through nanotechnology, these can be assembled into formulations with enhanced antimicrobial properties, high safety, and low toxicity. This review explores polysaccharides, including chitosan, alginate, starch, pectin, cellulose, hemicellulose, gums, carrageenan, dextran, pullulan, and hyaluronic acid, used in nanotechnology, highlighting their advantages and limitations as nanocarriers. Addressing the global urgency for alternative antimicrobials, we examined natural compounds derived from plants, microorganisms, and animals, such as phytochemicals, bacteriocins, animal antimicrobial peptides, and proteins. Focusing on their protection and retained activity, this review discusses polysaccharide-based nanoformulations with natural antimicrobials, including nanoparticles, nanoemulsions, nanocapsules, nanoplexes, and nanogels. Special emphasis is placed on strategies and formulations for the encapsulation, entrapment, and conjugation of natural compounds using polysaccharides as protective carriers and delivery systems, including a brief discussion on their future applications, prospects, and challenges in scaling up. Full article

Alginate is one of the most utilized biopolymers for the encapsulation of polyphenols throughout ionic gelation. For improvement in the encapsulation of polyphenols, other biopolymers and/or fillers can be employed. The purpose of this study was to include pullulan and/or disaccharides in an alginate encapsulation mixture to monitor whether we would achieve higher encapsulation of chokeberry juice polyphenols. Alginate hydrogel beads were used as controls, and through the results for total polyphenol and proanthocyanidin contents, concentrations of individual polyphenols, and antioxidant activities, it can be observed that pullulan and/or disaccharides had an impact on the encapsulation of these bioactives. Alginate/pullulan hydrogel beads had the highest contents of total polyphenols and proanthocyanidins (8.60 g/kg and 2.37 g/kg, respectively), whereas alginate/trehalose hydrogel beads had the lowest (5.50 g/kg and 1.16 g/kg, respectively). All hydrogel beads, except alginate/pullulan/sucrose, had higher anthocyanin (cyanidin-3-galactoside and cyanidin-3-arabinoside) contents than alginate beads (404.37 mg/kg and 89.97 mg/kg, respectively), but the most efficient combination for encapsulation of chokeberry anthocyanins was alginate/pullulan (477.32 mg/kg and 109.60 mg/kg, respectively). The highest concentration of neochlorogenic acid was determined in controls (260.14 mg/kg), while the highest concentration of chlorogenic acid in alginate/pullulan/sucrose beads (229.51 mg/kg). Quercetin-3-glucoside was evaluated as having the highest concentration in alginate/pullulan hydrogel beads (35.45 mg/kg). The data obtained through this study highlight the importance of the composition of an encapsulation mixture in order to achieve high encapsulation of chokeberry juice polyphenols. High encapsulation efficiency was obtained for anthocyanins, especially when pullulan was used in combination with alginate. Full article

Background: Self-administered orally dissolving films (ODFs) encapsulating inactivated influenza vaccines represent an effective strategy for stimulating mucosal immunity. While this vaccination method offers several advantages over conventional influenza vaccines, a comparative efficacy study remains lacking. Methods: Female BALB/c mice were immunized with inactivated A/PR/8/34 (H1N1) either via orogastric inoculation or through the oral mucosal delivery using pullulan and trehalose-based ODF vaccines. Each group received equivalent antigen doses across three immunizations. Humoral responses and antibody functionality were assessed using sera collected post-immunization. After lethal viral challenge, other immunological and virological parameters were determined in corresponding tissues. Body weight and survival were monitored over a 14-day period after challenge. Results: ODF vaccination elicited significantly higher virus-specific IgA levels, HAI titers, and neutralizing antibody activity than oral gavage. After the viral challenge, ODF-immunized mice exhibited stronger IgG and IgA responses in respiratory tissues, increased antibody-secreting cells in lungs and spleen, and elevated germinal center B cells and CD8⁺ T cell responses. Both vaccination methods reduced lung pro-inflammatory cytokines and provided full protection against lethal challenge; however, the ODF group showed lower cytokine levels, better weight maintenance, and reduced viral loads. Conclusions: ODF vaccination elicits more robust systemic and mucosal immune responses than oral vaccination and may serve as a promising alternative method of influenza vaccine delivery. Full article

Background: To enhance the bioavailability and neuroprotective efficacy of biatractylolide against Alzheimer’s disease by developing a novel Tween-80-modified pullulan–chenodeoxycholic acid nanoparticle as a delivery vehicle. Methods: Chenodeoxycholic acid (CDCA) was chemically conjugated to pullulan to yield hydrophobically modified pullulan (PUC), onto which polysorbate 80 (Tween-80) was subsequently adsorbed. The PUC polymers with CDCA substitution levels were analyzed by ¹H NMR spectroscopy. Nanoparticles were fabricated via the dialysis method and characterized by transmission electron microscopy and dynamic light scattering for morphology, size, and surface charge. In vitro neuroprotection was assessed by exposing SH-SY5Y and PC12 cells to 20 µM Aβ_25-35 to induce cytotoxicity, followed by pretreatment with biatractylolide-loaded PUC (BD-PUC) nanoparticle solutions at various biatractylolide concentrations. The in vivo brain-targeting capability of both empty PUC and BD-PUC particles was evaluated using a live imaging system. Results: The ¹H NMR analysis confirmed three distinct CDCA substitution degrees (8.97%, 10.66%, 13.92%). Transmission electron microscopy revealed uniformly dispersed, spherical nanoparticles. Dynamic light scattering measurements showed a hydrodynamic diameter of ~200 nm and a negative zeta potential. Exposure to 20 µM Aβ_25-35 significantly reduced SH-SY5Y and PC12 cell viability; pretreatment with BD-PUC nanoparticles markedly enhanced cell survival rates and preserved cellular morphology compared to cells treated with free biatractylolide. Notably, the cytoprotective effect of BD-PUC exceeded that of the free drug. In vivo imaging demonstrated that both empty PUC and Tween-80-adsorbed BD-PUC nanoparticles effectively accumulated in the brain. Conclusions: The protective effect of BD-PUC on SH-SY5Y and PC12 cells induced by Aβ_25-35 was higher than free biatractylolide solution, and the BD-PUC nanosolution modified with Tween-80 showed a brain-targeting effect. Full article

Microcirculation damage, dermal thickening, and difficulty in the spatiotemporal coordination of key platelet factor 4 (CXCL4) and transforming growth factor-β (TGF-β) contribute to the lack of effective treatments for systemic sclerosis (scleroderma, SSc). To address these challenges, we proposed a novel synergistic drug combination of ginsenoside Rk3 (CXCL4 regulator) and metformin (Met, TGF-β regulator) based on molecular docking and developed an ultra-long release, dual-target regulation hydrogel microneedle system (Rk3/Met URS MN). The rapidly dissolving tips of this hydrogel microneedle consisted of polyvinyl alcohol and polyvinylpyrrolidone, and were loaded with polydopamine-coated, coordination-induced self-assembled Rk3/Met nanomedicines. These micro-tips could spatiotemporally synchronize transdermal delivery of the hydrophobic Rk3 and hydrophilic Met, providing ultra-long release for up to 10 days with a single administration. The recombinant collagen CF-1552/oxidized pullulan-based (CAOP) hydrogel backing exhibited skin self-adhesiveness and excellent mechanical properties and could perform localized moisture retention and free radical scavenging at the lesion site. In vitro and in vivo efficacy studies, along with bioinformatics analysis of RNA sequencing, demonstrated that the Rk3/Met URS MN achieved immune modulation, anti-inflammatory effects, angiogenesis promotion, and antifibrosis in SSc through synergistic CXCL4/TGF-β dual-target regulation. Notably, on the 10th day, the dermal thickness decreased from 248.97 ± 21.3 μm to 152.7 ± 18.1 μm, with no significant difference from the normal group, indicating its significant potential in clinical applications in SSc. Full article

Carbohydrate-based biodegradable films offer an eco-friendly alternative to conventional petroleum-derived packaging for agricultural commodities. Derived from renewable polysaccharides such as starch, cellulose, chitosan, pectin, alginate, pullulan, and xanthan gum, these films exhibit favorable biodegradability, film-forming ability, and compatibility with food systems. This review presents a comprehensive analysis of recent developments in the preparation, functionalization, and application of these polysaccharide-based films for agricultural packaging. Emphasis is placed on emerging fabrication techniques, including electrospinning, extrusion, and layer-by-layer assembly, which have significantly enhanced the mechanical, barrier, and antimicrobial properties of these materials. Furthermore, the incorporation of active compounds such as antioxidants and antimicrobials has improved the performance and shelf-life of packaged goods. Despite notable advancements, key limitations such as moisture sensitivity, poor mechanical durability, and high production costs persist. Strategies including polymer blending, nanofiller incorporation, and surface modification are explored as potential solutions. The applicability of these films in packaging fruits, vegetables, dairy, grains, and meat products is also discussed. By assessing current progress and future prospects, this review underscores the importance of carbohydrate-based films in promoting sustainable agricultural packaging systems, reducing environmental impact through the advancement of circular bioeconomy principles and sustainable development. Full article

The objective of this study was to design an active packaging material based on a polyethylene terephthalate (PET)/polypropylene (PP) film modified with a pullulan coating enriched with 1, 5, and 10% of clove essential oil (CEO). The physical properties of modified PET/PP films, including opacity, UV, and light visible barrier properties, were evaluated, and calorimetric measurement of color (L*a*b*) was performed, followed by determination of their potential of antioxidant activity and antimicrobial properties against foodborne pathogenic bacteria (E. coli, S. Enteritidis, S. aureus and L. monocytogenes) were characterized. Additionally, the effectiveness of the active packaging in reducing pathogenic bacteria on spinach leaves was evaluated. The PET/PP film with a pullulan coating enriched with CEO did not affect their transparency. The incorporation of CEO improved the film’s UV and visible light barrier properties without causing noticeable color changes while also exhibiting good antioxidant and antimicrobial activity. Furthermore, the application of active packaging effectively inhibited the growth of pathogenic bacteria on spinach leaves, demonstrating its potential for food preservation. Full article

To learn about the yeast biodiversity of Hungarian honeys and to isolate osmotolerant yeasts, fifteen different honey varieties, beeswax, and bee bread were purchased, and samples of another, but highly osmotic material, tree sap (cherry, sour cherry, and plum), were collected from the northeastern region of the country. In total, 60 yeast strains were isolated and their taxonomic positions were determined by barcode sequences using ITS1-NL4 primers. The honey products contained mostly Zygosaccharomyces and Starmerella species. In addition, Hanseniaspora uvarum, Rhodotorula mucilaginosa and diobovata, Sporobolomyces roseus, Filobasidium magnum, Naganishia sp., and Aureobasidium pullulans were also present in smaller numbers. In contrast, tree saps contained Metschnikowia and Pichia fermentas cells. Further results suggest that some of the yeasts in honey can only “survive”, while others can propagate at high sugar levels, generally between 600 and 700 mg/g, with a predominance of fructose. Properties important for pathogenicity, such as invasive hyphae production, gelatin melting ability, and growth at 37 °C, were also examined. Hanseniaspora uvarum and Pichia fermentans representatives seemed to be negative for gelatin hydrolysis, while the other strains were able to melt gelatin. Although some of the strains could produce hyphae-like structures at 25 °C, none of them could grow at 37 °C. Full article

This review compares 32 studies (2000–2024) on plant- and microbial-based organic disease management to control grapevine pests and diseases. A systematic literature search provided 24 studies on microbial agents and 8 on plant treatments. Their effectiveness against key pathogens, including downy mildew, powdery mildew, and gray mold, was compared. Microbial agents such as Candida sake inhibited Botrytis cinerea by up to 80% in the lab and Pseudomonas sp. dramatically reduced grapevine lesion lengths by 32–52% in field conditions, while Bacillus subtilis reduced powdery mildew by 96% in greenhouse conditions and A. pullulans reduced Ochratoxin A infection by 99% in field conditions. In laboratory conditions, C. guilliermondii A42 reduced grape rot to 8–22% and A. cephalosporium B11 reduced it to 16–82%, confirming A42’s greater efficacy. Plant-derived agents and essential oils, including lavender and cinnamon, suppressed 100% of pathogens in vitro, whereas copper coupled with plant-derived agents reduced disease incidence by up to 92% under field conditions. While promising, plant-derived agents are plagued by formulation instability, which affects shelf life and effectiveness, while microbial agents must be kept under stringent storage conditions and can be variable under different vineyard conditions. These limitations identify the requirement for a stronger formulation strategy and large field validations. Organic disease management offers several important benefits, such as environmental safety, biodegradability, compatibility with organic cultivation, and low pesticide dependence. The application of these agents in pest management systems is ecologically balanced, improves soil health, and enables sustainable vineyard management. Full article

Topical approaches to dermatophytosis have the advantage of targeted therapy and minimal side effects and are patient-friendly. The present study focused on obtaining thin, flexible, and transparent bioadhesive polymeric films loaded with terbinafine hydrochloride (TH), in order to be administered to the skin affected by fungal infection. Polymeric films based on pullulan (P), oxidized pullulan (T-OP), sodium carboxymethylcellulose (NaCMC), and glycerin were obtained by the casting and evaporation technique, and the solubility of the drug was significantly increased by micellar solubilization with Tween-80, thus avoiding the use of organic solvents. Physico-chemical characterization through the FTIR technique and EDX analysis indicates the absence of strong interactions between the drug and the polymer, and the loading efficiency highlights the uniform distribution of the drug. The mechanical properties, bioadhesion, contact angle, and water sorption capacity highlight optimal adhesion parameters on the skin. In vitro studies indicate a prolonged drug release, in the first 300 min, of 80% and 60% for F2_TH and F1_TH, respectively, and the release kinetics follow the Weibull model. Significant antifungal activity was obtained for both polymeric films. The biocompatibility of the ingredients, the gentle technique for obtaining the films, and the results obtained from their analysis represent promise for their applicability in topical antifungal treatment. Full article

Monilinia spp., which causes brown rot, is one of the most damaging pathogens in stone fruits. Researchers are exploring epiphytic and endophytic microorganisms with the potential to suppress pathogens, control pathogenic microorganisms, and/or promote plant growth. In this study, microorganisms with antagonistic activity against three Monilinia species were isolated from plum orchard soil and plum fruits. Antagonism tests in vitro showed strong antagonistic properties of six strains of bacteria and two yeast-like fungi against M. fructigena, M. fructicola, and M. laxa, with growth inhibition from 45.5 to 84.6%. The antagonists were identified and characterized at the genetic level using whole genome sequencing (WGS). Genes involved in antibiotic resistance, virulence, secondary metabolite synthesis, and plant growth promotion were identified and characterized through genome mapping, gene prediction, and annotation. None of the microorganisms studied were predicted to be pathogenic to humans. The results of this study indicate that the bacteria Bacillus pumilus, B. velezensis, two strains of Lysinibacillus agricola, Pseudomonas chlororaphis isolated from stone fruit orchard soil, and the yeast-like fungus Aureobasidium pullulans, isolated from plums, are promising candidates for the biological control of Monilinia spp. Full article

This study aimed to investigate the endophytic microbial populations associated with wild and domesticated grapevines using both culture-based and culture-independent methods. Through culture-based methods, 148 endophytes were identified. The dominant fungal species included Aureobasidium pullulans, Alternaria alternata, and Cladosporium allicinum, while predominant bacterial species were Ralstonia pikettii, Nocardia niigatensis, and Sphingomonas echinoides. Culture-independent methods employed metagenomic techniques to explore microbial biodiversity, focusing on targeted amplification of bacterial 16S rRNA as well as fungal ITS and 26S rRNA gene regions. The main bacterial species identified included Halomonas sp., Sphingomonas sp. and Massilia sp., whereas the fungal population was dominated by Cladosporium sp., Malassezia sp. and Mucor sp. The findings revealed that vineyard management practices did not lead to statistically significant variations in microbial communities. The consistent presence of these genera across all samples suggests that they are stable components of the grapevine endophytic microbiota, remaining relatively unaffected by external environmental factors. Full article

Polysaccharide-based hydrogels have emerged as indispensable materials in tissue engineering and wound healing, offering a unique combination of biocompatibility, biodegradability, and structural versatility. Indeed, their three-dimensional polymeric network and high water content closely resemble the natural extracellular matrix, creating a microenvironment for cell growth, differentiation, and tissue regeneration. Moreover, their intrinsic biodegradability, tunable chemical structure, non-toxicity, and minimal immunogenicity make them optimal candidates for prolonged drug delivery systems. Notwithstanding numerous advantages, these polysaccharide-based hydrogels are confronted with setbacks such as variability in material qualities depending on their source, susceptibility to microbial contamination, unregulated water absorption, inadequate mechanical strength, and unpredictable degradation patterns which limit their efficacy in real-world applications. This review summarizes recent advancements in the application of polysaccharide-based hydrogels, including cellulose, starch, pectin, zein, dextran, pullulan and hyaluronic acid as innovative solutions in wound healing, drug delivery, tissue engineering, and regenerative medicine. Future research should concentrate on optimizing hydrogel formulations to enhance their effectiveness in regenerative medicine and antimicrobial therapy. Full article