Search Results (317)

We report a novel tablet-based reverse titration system for rapid, point-of-use measurement of water hardness, overcoming key limitations of conventional EDTA titration. Reagents are encapsulated in pullulan matrix giving two separate tablets. The first tablet contains the Eriochrome black T (EBT) and N-cyclohexyl-3-aminopropanesulfonic acid (CAPS) buffer, while the second encapsulates ethylenediaminetetraacetic acid (EDTA) disodium salt dihydrate. The system employs a trimodal detection strategy: qualitative screening via immediate color change with the EBT tablet, semi-quantitative estimation through combined tablet dissolution and adjusting the sample volume to a reference level, and quantitative determination using reverse titration, where water is gradually added until the red wine endpoint appears. This approach enhances interference tolerance from competing metal ions and improves accuracy over traditional methods. Testing with real water samples showed excellent agreement with standard titration. The tablets remain stable for over seven months, and the system eliminates the need for skilled personnel, laboratory equipment, or bulky instrumentation. This low-cost, user-friendly, and interference-tolerant platform enables rapid and accurate water hardness assessment at the point of use. Full article

This review consolidates recent advancements in microbial biotechnology for sustainable food systems. It focuses on the fermentation processes used in this sector, emphasizing precision fermentation as a source of innovation for alternative proteins, fermented foods, and applications of microorganisms and microbial bioproducts in the food industry. Additionally, it explores food preservation strategies and methods for controlling microbial contamination. These biotechnological approaches are increasingly replacing synthetic additives, contributing to enhanced food safety, nutritional functionality, and product shelf stability. Examples include bacteriocins from lactic acid bacteria, biodegradable microbial pigments, and exopolysaccharide-based biopolymers, such as pullulan and xanthan gum, which are used in edible coatings and films. A comprehensive literature search was conducted across Scopus, PubMed, ScienceDirect, and Google Scholar, covering publications from 2014 to 2025. A structured Boolean search strategy was applied, targeting core concepts in microbial fermentation, bio-based food additives, and contamination control. The initial search retrieved 5677 articles, from which 370 studies were ultimately selected after applying criteria such as duplication removal, relevance to food systems, full-text accessibility, and scientific quality. This review highlights microbial biotransformation as a route to minimize reliance on synthetic inputs, valorize agri-food byproducts, and support circular bioeconomy principles. It also discusses emerging antimicrobial delivery systems and regulatory challenges. Overall, microbial innovations offer viable and scalable pathways for enhancing food system resilience, functionality, and environmental stewardship. Full article

Edible composite coatings represent an alternative approach to reducing postharvest losses and extending the shelf life of perishable fruits. This study developed a nano-biopolymer coating by integrating pullulan (PUL), nano-silica (Nano-SiO₂), and tea polyphenols (TP) to retard deterioration in cherry tomatoes (Solanum lycopersicum var. cerasiforme). Optimized through response surface methodology (0.06% Nano-SiO₂, 0.1% TP, 1.8% PUL, 0.77% glycerol), the resulting Nano-SiO₂/PUL/TP composite film showed improved barrier properties (water vapor permeability, WVP: 0.2063 g·mm·m⁻²·h⁻¹·kPa⁻¹) and increased mechanical strength (tensile strength, TS: 2.62 MPa; elongation at break, EB: 67.67%), which may be attributed to a homogeneous microstructure stabilized via intermolecular hydrogen bonding. The composite coating exhibited significant (p < 0.05) antioxidant activity (59.04% DPPH·scavenging) compared to the PUL film (1.17%) and showed efficacy against S. aureus. When applied to cherry tomatoes stored at 4 °C for 15 days, the coating contributed to improved postharvest quality by reducing weight loss (−27.6%) and decay incidence (−32.3%), delaying firmness loss (2.40 vs. 0.54 N in uncoated group, CK), suppressing respiration rate (−38.8%), and enhancing the retention of total acidity (+9.7%), vitamin C (+49.6%), and total soluble solids (+48.6%) compared to the CK (p < 0.05). Principal component analysis supported sensory evaluation results, indicating the coating helped maintain sensory quality (scores > 6.0) and commercial value while extending shelf life from 9 to 15 days. These results suggest that the Nano-SiO₂/TP/PUL composite coating may serve as a preservative for extending the shelf-life of cherry tomatoes by effectively reducing decay and mitigating quality degradation. Full article

Background/Objectives: The design of scaffolds that mimic the extracellular matrix has gained increasing attention in regenerative medicine. This study aims to develop and characterize electrospun nanofibrous scaffolds based on pullulan blended with either gelatin or gliadin and doped with selenium nanoparticles (Se NPs), to assess the influence of protein type and Se NP doping on scaffold performance and regenerative potential. Methods: Se NPs were synthesized via redox reaction and stabilized using pullulan. Electrospun scaffolds were then prepared by blending pullulan-stabilized Se NPs with either gelatin or gliadin. The resulting fibers were characterized using a multidisciplinary approach, including physicochemical (morphology, fiber dimension, swelling capacity, surface zeta potential, mechanical properties) and preclinical properties (antioxidant properties, fibroblast adhesion and proliferation, collagen expression). Results: Protein type influenced fiber morphology and dimensions, as well as mechanical behavior, with gelatin-based scaffolds demonstrating smaller fiber diameters and higher mechanical properties. The doping with Se NPs enhanced scaffold antioxidant properties without affecting fiber formation. Moreover, all scaffolds supported fibroblast proliferation, but those containing Se NPs showed enhanced modulation of ECM gene expression. Conclusions: The results show that scaffolds doped with Se NPs exhibited superior performance compared to the undoped counterparts, offering promising platforms for chronic wound reparation. Full article

This study characterised the diversity of filamentous fungi and yeasts during Malbec grape fermentation in the Southern Oasis of Mendoza (Argentina) winegrowing region, under different sanitary conditions and SO₂ treatments, using morphological and ITS-RFLP-based molecular methods. Alternaria, Cladosporium and Penicillium were present in both sound and damaged grapes, while Aspergillus and Botrytis were primarily found in damaged grapes. The predominant yeast species in both sound and damaged grape must, at lower and higher maturity levels, were Aureobasidium pullulans and Hanseniaspora spp. At higher grape ripening levels species diversity increased, with Hanseniaspora vineae, Metschnikowia pulcherrima and Candida membranifaciens dominating, and others such as Pichia kudriavzevii and Issatchenkia terricola appearing. A. pullulans and M. pulcherrima were highly tolerant to SO₂. Notably, the species Meyerozyma guilliermondii, Zygoascus hellenicus and Hanseniaspora uvarum were exclusively present in damaged grape must, while Zygosaccharomyces bailii was also found in sound grape must. Hanseniaspora spp. and P. kudriavzevii predominated at mid-fermentation and persisted at the end of the process, highlighting their resistance to wine conditions and their potential to influence post-fermentative dynamics. These findings emphasise the significant influence of grape sanitary status on mycobiota composition, with important implications for fermentation behaviour and final wine quality. Full article

A study was carried out to identify the filamentous fungi and yeasts present in rotten wine grapes from two subzones of the Southern oasis of Mendoza winegrowing region, to assess the occurrence of tenuazonic acid (TA), a mycotoxin produced by the Alternaria genus, and to evaluate the wine spoilage potential of the associated yeasts in vitro and during microvinifications. The main fungal genera present were Alternaria (69.3%), followed by Aspergillus (16.8%), Penicillium (9.3%), and Cladosporium (4.6%), while the dominant yeast species Metschnikowia pulcherrima (23.1%), Aureobasidium pullulans (20.2%) and Hanseniaspora uvarum (13.0%) were followed by H. vineae (11.6%), Zygosaccharomyces bailii (10.4%), and H. guilliermondii (9.2%). Additionally, 94.1% of the rotten samples were contaminated with TA, with the highest level found in the Cabernet Sauvignon variety. No geographic association was found in the incidence of the different fungal genera or yeast species, nor in the occurrence of TA. Almost all of the tested yeasts produced H₂S, the majority of the Hanseniaspora strains produced acetic acid, and only one M. pulcherrima strain produced off-flavours in in vitro tests. Wines co-fermented with H. uvarum L144 and S. cerevisiae showed higher volatile acidity and lower fruity aroma and taste intensity. Therefore, processing bunch rot could pose a toxicological and microbiological risk to winemaking due to the high incidence of Alternaria and TA, as well as the potential of the associated yeasts to spoil wine. Full article

Immunoregulation is an emerging treatment strategy to promote wound healing by modulating the local immune system at the wound site. In this study, an extracellular matrix biomimetic and polysaccharide-based hydrogel was engineered to regulate the wound immune environment through Michael-type addition between maleimidyl pullulan and chitosan modified with hydroxyproline. The proposed hydrogel exhibited favorable injectable and self-healing properties, which facilitated the full coverage of irregularly shaped wounds. A natural polyphenol, epigallocatechin-3-gallate (EGCG), was incorporated into hydrogels, which thereby exhibited excellent biocompatibility, good reactive oxygen species (ROS) scavenging ability, anti-inflammatory activity, and antibacterial properties against S. aureus and E. coli. Furthermore, evaluations of a full-thickness skin defect mice model showed that the hydrogel with EGCG effectively alleviated the inflammatory response by reducing pro-inflammatory cellular infiltration and down-regulating the inflammatory cytokine TNF-α, while up-regulating anti-inflammatory cytokine IL-10. Notably, a faster wound healing rate was also achieved by the better promotion effect of the hydrogel on increasing the formation of re-epithelialization, granulation tissue generation, collagen deposition, and angiogenesis. Therefore, our immunoregulatory strategy showed great potential in the design of biomaterials for wound management. Full article

Microbial enzymes, due to their efficiency, specificity, and sustainability, are central to innovative biotechnological strategies aimed at optimizing industrial processes such as winemaking. In this study, the potential of Aureobasidium pullulans m11-2, a native dimorphic fungus from the wine ecosystem, was evaluated as a source of hydrolytic enzymes capable of degrading grape cell wall polysaccharides. The strain was identified at the molecular level and characterised in terms of its morphology. To maximise enzyme production, various culture media were tested. Among the concentrations tested, the optimal levels of glucose and pectin were 1 g L⁻¹ and 10 g L⁻¹, respectively. The partially constitutive and inducible nature of the various polysaccharidase activities (pectinases, cellulases, and xylanases) was confirmed. The effect of grape skins (a winemaking by-product) on microbial growth and enzyme synthesis was evaluated, achieving a pectinase activity of 0.622 U mL⁻¹ when combined with 1 g L⁻¹ of glucose. Maximum enzyme yields were detected during the exponential growth phase in both citrus pectin and grape skin media, suggesting favorable conditions for continuous bioprocessing. These results confirm that A. pullulans m11-2 is an interesting microbial option for producing polysaccharidases that can be adapted to sustainable production systems. Full article

Background/Objectives: Anaerobic gut fungi (Neocallimastigomycota) are biotechnologically relevant, lignocellulose-degrading microbes with under-explored biosynthetic potential for secondary metabolites. Untargeted metabolomic profiling with gas chromatography–mass spectrometry (GC-MS) was applied to two gut fungal strains, Anaeromyces robustus and Caecomyces churrovis, to establish a foundational metabolomic dataset to identify metabolites and provide insights into gut fungal metabolic capabilities. Methods: Gut fungi were cultured anaerobically in rumen-fluid-based media with a soluble substrate (cellobiose), and metabolites were extracted using the Metabolite, Protein, and Lipid Extraction (MPLEx) method, enabling metabolomic and proteomic analysis from the same cell samples. Samples were derivatized and analyzed via GC-MS, followed by compound identification by spectral matching to reference databases, molecular networking, and statistical analyses. Results: Distinct metabolites were identified between A. robustus and C. churrovis, including 2,3-dihydroxyisovaleric acid produced by A. robustus and maltotriitol, maltotriose, and melibiose produced by C. churrovis. C. churrovis may polymerize maltotriose to form an extracellular polysaccharide, like pullulan. GC-MS profiling potentially captured sufficiently volatile products of proteomically detected, putative non-ribosomal peptide synthetases and polyketide synthases of A. robustus and C. churrovis. The triterpene squalene and triterpenoid tetrahymanol were putatively identified in A. robustus and C. churrovis. Their conserved, predicted biosynthetic genes—squalene synthase and squalene tetrahymanol cyclase—were identified in A. robustus, C. churrovis, and other anaerobic gut fungal genera. Conclusions: This study provides a foundational, untargeted metabolomic dataset to unmask gut fungal metabolic pathways and biosynthetic potential and to prioritize future efforts for compound isolation and identification. Full article

The present study develops and optimizes a jet-cutting encapsulation method using a laboratory-scale encapsulator to incorporate herbal dietary supplements into fermented milk products. Sodium alginate and pullulan were selected as core and coating polymers, respectively, after rheological screening demonstrated that 1% alginate (η ≈ 350–450 Pa·s at 22–25 °C) and 2% pullulan (η ≈ 400 Pa·s at 25–30 °C) provide a balance between atomization, shell integrity, and fluidity. Under optimized conditions, capsules of 1.00 ± 0.05 mm diameter and high sphericity (aspect ratio 1.08 ± 0.03) were produced. In vitro gastrointestinal simulation confirmed capsule stability in simulated gastric fluid (pH 2.0) and complete disintegration within 120 min in simulated intestinal fluid (pH 7.2). Inclusion of 8% (w/w) capsules in a fermented milk beverage preserved appearance, texture, flavor, and color while increasing viscosity from 2.0 to 4.0 Pa·s. Titratable acidity rose from 87 °T at 24 h to 119 °T at 120 h, with sensory quality remaining acceptable; substantial gas formation and excessive sourness occurred only after 168 h, defining a 5-day refrigerated shelf life. These findings demonstrate that the 1% alginate–pullulan capsule system successfully protects plant extracts during gastric transit and enables targeted intestinal release, while maintaining the sensory and rheological properties of the fortified fermented milk product. 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

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