Search Results (244)

Polyvinyl alcohol (PVA) hydrogels modified through radical polymerization under UV irradiation and Ce⁴⁺ ion treatment were investigated as a potential platform for developing highly sensitive biosensors for rapid biochemical oxygen demand analysis in water. These modifications enhance PVA physicochemical properties, including mechanical strength, stability, and biocompatibility, making it promising for immobilizing microorganisms in bioanalytical systems. A dual-mediator biosensor system using ferrocene (FC) and neutral red (NR) was developed with yeast Blastobotrys adeninivorans immobilized in modified PVA. The FC+NR–B. adeninivorans–PVA–Ce⁴⁺ system exhibited high sensitivity (linear range of 0.1–3.81 mgO₂/dm³), selectivity, and operational stability (up to 37 days service life), outperforming existing analogs. Testing with wastewater confirmed strong correlation with standard BOD₅, highlighting the potential for monitoring water quality. The described radical modification method is a simple and effective approach for creating sensitive and stable biosensors. It opens up new possibilities for environmental monitoring technology. Full article

Objective: The objective of our study was to synthesize and characterize silica–copper nanomaterials and to evaluate their biological properties (antibacterial, redox, cytotoxic, and ecotoxic) for potential applications. Methods and Results: Si/Cu-based materials were prepared by a sol–gel method. They were characterized by XRD, UV-Vis, and SEM-EDS. The antibacterial activity of the materials was evaluated against Gram-positive bacteria (Staphylococcus aureus, Bacillus cereus), Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium), and yeasts (Candida albicans, Saccharomyces cerevisiae). The nanomaterial that was calcined at 500 °C exhibited greater antibacterial efficacy compared to the gel form. S. typhimurium demonstrated the highest susceptibility, whereas S. aureus and P. aeruginosa were the most resistant of the tested bacteria. Both yeasts exhibited comparable sensitivity (MBC = 1.0 mg/mL). The redox activity of both nanomaterials was tested at pH 7.4 (physiological) and 8.5 (optimal) by the activated chemiluminescent method. The nanocomposites significantly inhibited the free-radical and ROS generation. This presents them as redox regulators in living systems. The cytotoxic effects in normal BEAS-2B and tumor A549 human cell lines were assessed microscopically and by the cell viability neutral red uptake assay, CC₅₀ being evaluated. The observed effects suggest moderate, similar cytotoxicity in both cell lines. The ecotoxicity study using Daphnia magna showed an LC₅₀ of ~7–8 mg/L about Si/Cu/500. The LC₅₀ for Si/Cu (gel) was lower than 0.25 mg/L, indicating an increase in toxicity with increased exposure time. Conclusions: Possible applications of the newly synthesized nanomaterials include antimicrobial coatings, drug delivery systems, antioxidant additives in various formulations, and water purification. Full article

Perfluorooctanesulfonic acid (PFOS) and perfluorooctanoic acid (PFOA) pose significant health risks through various exposure pathways, including ingestion of contaminated food and water, as well as dermal absorption. Aquatic organisms are especially at risk, as water bodies serve as primary pathways for the transport and transformation of these chemicals. While the biodegradation study was previously performed using a bacterial consortium from the activated sludge compartment at Zeekoegat WWTP, the ecotoxicological implications of the treated effluents remained unclear, particularly given the potential presence of degradation products. To address this gap, the present study used bioassays to evaluate the acute toxicity and endocrine-disrupting potential of PFOS and PFOA. For this purpose, PFOS and PFOA concentrations ranged from 58 ng/L to 1050 ng/L, and two types of bioassays were used: the Daphnia magna acute toxicity test, which examined the short-term lethal effects of the samples on a small freshwater organism (Daphnia magna), and the Yeast Estrogen Screen (YES), which measured estrogenic activity, an important indicator of potential endocrine disruption. Results revealed detectable estrogenic activity at environmentally relevant concentrations, with PFOS showing higher activity than PFOA. The estradiol equivalency (EEQ) values in samples containing PFOA ranged from 0.23 ± 0.029 ng/L to 3.15 ± 0.056 ng/L and from 0.43 ± 0.036 ng/L to 1.96 ± 0.086 ng/L in samples containing PFOS. Daphnia magna bioassays showed 100% mortality in samples containing PFOS at concentrations ≥ 62 ng/L and in samples containing PFOA at concentrations ≥ 142 ng/L, classifying them as ‘Very High Acute Hazard’ falling into Hazard Class V (100% mortality) according to the classification system proposed in 2003 by Persoone and co-workers. These bioassays helped to determine whether the degradation products were more toxic compared to the parent compounds, thereby supporting the objective of this study to assess environmental safety post-treatment. Full article

The identification and development of novel antimicrobials is a crucial challenge in the face of increasing antibiotic and antimycotic resistance. As such, there is growing interest in exploring the chemical diversity of natural sources, such as invasive seaweeds such as Asparagopsis armata. The valorization of such sources can further contribute to the development of bio-based industries, aligning with societal goals for environmental and economic sustainability. Therefore, a solid-liquid extraction method was performed using ethanol, and the obtained extract was studied for chemical composition elucidation, bioactivity, and toxicity evaluation. Analysis by GC-MS revealed some major chromatographic peaks, including floridoside (2-α-O-D-galactopyranosyl glycerol), glycerol, and oleamide. Also, several other smaller peaks were tentatively attributed to Low Molecular Weight Carbohydrate Derivatives, including isosaccharino-1,4-lactone, which had only been reported once in nature. The extract demonstrated significant antioxidant activity as measured by Ferric Reducing Antioxidant Potential and Oxygen Radical Absorption Capacity, but not by Lipid Peroxidation Inhibitory Potential assays, which is in line with its polar nature. The extract exhibited antimicrobial activity against various microorganisms, with a MIC of 2 mg/mL observed for Staphylococcus epidermidis, Vibrio parahaemolyticus, and the three yeast strains tested. Moreover, the extract inhibited the growth and phenotypic changes in filamentous fungi, which may result in reduced virulence. Specifically, the extract inhibited sporulation in Aspergillus fumigatus and orange pigmentation in Fusarium graminearum, possibly by a reduction in the production of aurofusarin, rubrofusarin, and mycotoxins. In vitro cell viability assays in 3T3, RAW264.7, and HaCaT demonstrated the extract was not cytotoxic or presented low cytotoxicity at concentrations up to 0.1 mg/mL, but a strong cytotoxic effect was observed at 1 mg/mL. At non-cytotoxic concentrations, the ethanol extract inhibited up to 48% of NO production in LPS-stimulated RAW264.7. This may indicate that anti-inflammatory activity may add to the antimicrobial activity in human and animal systemic and topical applications of the extract. In this work, new molecules were reported in A. armata, and the bioactivities reported were novel for this extract and algal species—especially through the choice of uncommon but very relevant pathogens to study. Our findings are a valuable contribution to addressing challenges in human and animal health, food and feed technology, as well as animal husbandry and agriculture. Full article

Lycopene, a natural carotenoid with antioxidant and health-promoting properties, has attracted attention as a valuable compound for the food, pharmaceutical, and cosmetic industries. Conventional production methods based on plant extraction or chemical synthesis are limited by low yields, high costs, and environmental concerns. In this study, the oleaginous yeast Yarrowia lipolytica was engineered as an alternative microbial cell factory for sustainable lycopene biosynthesis using short-chain fatty acids (SCFAs)—such as acetate, butyrate, and propionate—as inexpensive, renewable carbon sources. Four heterologous genes from Pantoea agglomerans (crtI, crtB, crtE, and idi) were codon-optimized and integrated into the Y. lipolytica genome using different expression systems, including the Golden Gate Assembly strategy. Among the tested strains, PS05/4lyc/GGA, characterized by enhanced phospholipid biosynthesis, demonstrated the highest lycopene yield of 462.9 mg/g dry cell weight and a titer of 3.41 g/L on butyrate medium—values comparable to or exceeding those reported for bioreactor-scale fermentations. The results indicate that co-activation of phospholipid and carotenoid biosynthesis pathways creates favorable intracellular conditions for hydrophobic pigment accumulation. Moreover, the use of SCFAs improved acetyl-CoA availability and redirected carbon flux through the mevalonate pathway, enhancing productivity. Strains with elevated membrane lipid biosynthesis also exhibited higher metabolic stability and stress tolerance. Full article

The microbiota associated with Vanilla planifolia grown in three production systems in Puebla, México, was evaluated: shade cloth, cocuite, and acahual. Rhizosphere and soil samples were analyzed, from which bacteria, fungi, yeasts, and actinomycetes were isolated. The bacterial and actinomycete isolates were characterized morphologically and biochemically, and their potential as growth promoters was evaluated. Morphological and microscopic characteristics identified the fungi. In parallel, agronomic variables were measured in five plants per system, and the data were analyzed using ANOVA and Tukey’s test (p ≤ 0.05). The results showed that the shade cloth favored a greater number of internodes, total leaves, and biomass, although with a higher incidence of diseased leaves. The cocuite presented intermediate values, while the acahual had lower leaf density but fewer leaf health problems. Microbial composition varied across systems, with potentially beneficial bacteria and actinomycetes, as well as both beneficial and pathogenic fungi, being prominent. These findings demonstrate the influence of the management system on the microbiota and health of V. planifolia, providing a basis for more sustainable production strategies for vanilla cultivation in Mexico. Full article

The water sector faces a dual challenge: reducing energy consumption and carbon footprint while improving wastewater treatment efficiency. Anaerobic digestion (AD) remains the primary method for energy recovery in wastewater treatment plants (WWTPs). To enhance methane production and move toward carbon neutrality, co-digestion of sewage sludge with external substrates is gaining attention. This study evaluated nine organic substrates for their methane potential using the standardized Automatic Methane Potential Test System (AMPTS). The highest methane yield was obtained from sediment from a wine tank, reaching 1387 NmL CH₄/g VS, followed by yeast slurry, with 524 NmL CH₄/g VS. These values were over 6 and 2.5 times higher, respectively, compared to the methane potential of conventional mixed municipal sludge. Apple pomace, whey, food biowaste, and herbal maceration waste showed moderate improvements (301–388 NmL CH₄/gVS). When considering methane yield per gram of wet substrate, herbal maceration waste was the most efficient. A techno-economic analysis revealed that this substrate consistently achieved a net-positive energy balance (up to 170%) in large WWTPs, even at transport distances of 50 km. Other substrates also showed high potential, covering nearly 100% of energy demand under optimal conditions. In contrast, whey showed limited applicability due to transport constraints. These findings highlight the importance of substrate selection, particularly in practical efforts aimed at achieving energy self-sufficiency in wastewater treatment plants. It also provides WWTP operators with clear and practical insights into enhancing methane yields from anaerobic digesters while minimizing the risk of process inhibition. Full article

This study investigated the antimicrobial efficacy of supercritical carbon dioxide (SC-CO₂) plant extracts as a natural preservative, prolonging food shelf-life. The research evaluated the performance of 10 different extracts, including cinnamon, thyme, clove, and dashi, in low-fat food matrices. The results showed that these extracts significantly prolonged the shelf life of a plant-based and animal-based matrixes, with cinnamon and dashi extracts proving highly effective in plant-based matrix against mould and yeast growth for up to 65 days. A key part of the study focused on the interaction between these lipophilic extracts and black soldier fly larvae (BSFL) fat as a potential carrier system. While fats were expected to improve the extracts’ sensory properties and act as a delivery system, in vitro tests revealed an antagonistic effect. The lipophilic nature of the extracts’ active compounds caused them to be trapped within the fat phase, rendering them unavailable to interact with pathogens. These findings highlight the challenges and potential of using lipophilic natural antimicrobials in food systems and underscore the need for new strategies to optimize their efficacy. Full article

The increasing interest in natural preservatives has driven the search for effective microbial agents capable of controlling spoilage molds in cured meat products. In this study, the efficacy of Debaryomyces hansenii strains as biocontrol agents against spoilage molds in dry-cured meat products was evaluated through a dual experimental approach using both synthetic (PDA) and meat-derived media (LBM). While all D. hansenii strains demonstrated strong antifungal activity in nutrient-rich synthetic media, their performance in meat-like conditions was moderate to high, with significant differences depending on the mold species, the yeast strain, and their interaction with the culture medium. Our results highlight that antifungal efficacy is strongly influenced by the growth environment, underscoring the limitations of traditional in vitro assays that depend solely on synthetic media. Incorporating food-mimicking systems early in the screening process proved critical to identify strain–mold–medium combinations with the highest potential. These findings support the potential application of native D. hansenii strains as natural preservatives to enhance the safety and shelf life of dry-cured meats, emphasizing the importance of testing in conditions that closely resemble the target food environment to select the most effective biocontrol solutions. Full article

This study investigates the impact of light exposure on the biotransformation of chalcones in yeast cultures. 4′-Hydroxychalcones, with a hydroxyl group in the A-ring, are characteristic substrates efficiently converted into 4′-hydroxydihydrochalcones—compounds naturally occurring in medicinal plants such as Glycyrrhiza glabra (licorice), Stevia rebaudiana, and Angelica keiskei (ashitaba). These compounds are valued for their bioactivity and are relevant to natural product research. In this research, we present the outcomes of the selective microbial reduction of chalcones to dihydrochalcones using the yeast Yarrowia lipolytica KCh 71, cultivated under both light and dark conditions. The aim was to determine whether light exposure affects the efficiency or selectivity of the transformation. Furthermore, the effect of substrate photoisomerisation induced by light was investigated, as the trans–cis isomerisation of chalcones may affect their availability and affinity toward enzymatic systems. The resulting metabolites were analysed using chromatographic and spectroscopic methods. No significant differences in transformation efficiency were observed between light and dark conditions. In all tested conditions, the 4′-hydroxydihydrochalcones were obtained with high yield, typically exceeding 90% conversion. Additionally, the selective bioreduction of the α,β-unsaturated bond in selected 4′-hydroxychalcones by the studied yeast culture is an exceptionally efficient process. The primary factor influencing the reaction rate is the structure of the substrate, particularly the number and distribution of methoxyl groups on the B-ring. In addition, we establish biocatalytic access to three target dimethoxy dihydrochalcones—4′-hydroxy-2,4-dimethoxydihydrochalcone (5a), 4′-hydroxy-2,5-dimethoxydihydrochalcone (6a), and 4′-hydroxy-3,5-dimethoxydihydrochalcone (7a)—under mild conditions using Yarrowia lipolytica KCh 71. Under preparative-scale conditions (7-day incubation), a minor additional product (≤10%) was detected only for the 4′-hydroxy-2,5-dimethoxydihydrochalcone transformation and identified as 4′,5-dihydroxy-2-methoxydihydrochalcone (6b); no such side reaction was observed in short-term experiments. Full article

Background/Objectives: Receptor-mediated transcytosis utilizing the native transporters at the blood–brain barrier (BBB) is a growing strategy for the delivery of therapeutics to the brain. One of the major challenges in identifying appropriate human transcytosis targets is that there is a species-specific transporter expression profile at the BBB, complicating translation of successful preclinical candidates into humans. In an effort to overcome this obstacle and identify proteins capable of binding human-relevant BBB ligands, we generated and screened a BBB-targeting library against human-induced pluripotent stem cell-derived brain microvascular endothelial-like cells (iPSC-derived BMEC-like cells). As targeting molecules, we used lamprey antibodies, known as variable lymphocyte receptors (VLRs), and generated a VLR library by immunizing lamprey with iPSC-derived BMEC-like cells, and inserting the resultant VLR repertoire into the yeast surface display system. Methods: The yeast displayed VLR library was then panned against human iPSC-derived BMEC-like cells and lead VLRs were validated using human in vitro models and mouse and human ex vivo brain tissue sections. Results: Finally, brain uptake for a set of VLRs was validated in mice. Of the 15 lead VLR candidates, 14 bound to human BBB antigens, and 10 bound to the murine BBB. Pharmacodynamic testing using the neuroactive peptide neurotensin indicated that the lead candidate, VLR2G, could cross the mouse BBB after intravenous injection and deliver sufficient neurotensin payload to generate a pharmacological response and lower systemic body temperature. Conclusions: Together, these results demonstrate the application of a novel screening technique capable of identifying a VLR with human relevance that can cross the BBB and deliver a payload. Full article

Yeasts of the Candida genus are part of the normal human microbiota but can cause infections (candidiasis) under certain conditions. While Candida albicans remains the most common etiological agent, the prevalence of non-albicans Candida species—such as C. glabrata, C. tropicalis, C. krusei, C. parapsilosis, C. kefyr, C. lusitaniae, and the emerging multidrug-resistant C. auris—has been increasing. Effective treatment of candidiasis requires rapid and accurate identification of the causative species, particularly due to species-specific antifungal agent resistance patterns. The aim of this study was to evaluate the usefulness of five chromogenic media for the differentiation of Candida species: BD CHROMagar Candida (Becton Dickinson), CHROM ID Candida (bioMérieux), CHROMAgar Candida Plus (CHROMAgar France, Biomaxima), CHROMAgar Candida Plus (GRASO Biotech), and Brilliance Candida Agar (OXOID). A total of 175 strains from the following species were tested: C. albicans, C. parapsilosis, C. dubliniensis, C. lusitaniae, C. tropicalis, C. glabrata, C. kefyr, C. krusei, and C. auris. Species identification was confirmed by MALDI-TOF mass spectrometry using the MALDI Biotyper system (Bruker). Colony morphology, especially color characteristics, was assessed on each medium. The morphological features of most Candida species were consistent with the manufacturer’s descriptions and allowed for presumptive species-level identification. However, some species showed reproducible but previously undescribed morphological traits, including variations in colony shade. Notably, C. auris could not be reliably identified using BD, bioMérieux, or OXOID media. In conclusion, while chromogenic media are a helpful preliminary diagnostic tool, subtle differences in colony coloration can complicate interpretation. Diagnostic caution is recommended, and confirmatory methods such as MALDI-TOF remain essential for reliable identification, especially for emerging or less common Candida species. Full article

Considering the clinical relevance of commensal yeasts (Malassezia and Candida) and zoophilic dermatophytes (Microsporum canis and Trichophyton mentagrophytes) in dogs and cats, this study determines the prevalence of fungal species involved in ear and superficial skin infections in dogs and cats in Grenada and examines their antifungal susceptibility. The etiological agents were isolated from ear, skin, and hair samples of suspected clinical fungal cases using Sabouraud Dextrose Agar (SAB). The isolates’ identification comprised morphological, biochemical, and molecular methods encompassing micro-/macroscopy analysis. Biochemically, yeast isolates were identified by the BD Phoenix M50 microbial identification system, and additional validation of all fungal isolates was performed by polymerase chain reaction (PCR) and sequencing of the ITS region. Furthermore, the E-Test (Epsilometer Test) was used to determine the susceptibility patterns for four azole drugs: ketoconazole, itraconazole, fluconazole, and voriconazole. A total of 405 samples (266 ear, 61 skin, and 78 hair) were collected from 136 dogs and 43 cats. The identified species were Malassezia pachydermatis, Candida tropicalis, and Trichophyton spp. All isolates demonstrated (100%) resistant activity to fluconazole. Importantly, this knowledge will significantly contribute to our understanding of the epidemiology of fungal infections as well as provide guidelines for preventive measures against fungal infections in Grenada. Full article

Background/Objectives: Glucan particles (GPs), derived from Saccharomyces cerevisiae yeast, possess unique biomedical properties. Nevertheless, it is imperative that a comprehensive risk assessment is conducted during pre-clinical development. GPs are primarily constituted of a naturally occurring polymer known as β-glucan. This study characterized GPs, focusing on physicochemical attributes, biocompatibility, and immunomodulatory potential. Methods: GPs were characterized for size, morphology, surface charge, and protein encapsulation efficiency using dynamic light scattering (DLS), electron microscopy, and encapsulation assays. Biocompatibility was assessed through cytotoxicity assays (MTT), hemolysis tests, and measurement of reactive oxygen (ROS) and nitric oxide (NO) production in immune cells. Immunomodulatory potential was evaluated by cytokine and chemokine secretion analysis in peripheral blood mononuclear cells (PBMCs) and monocyte-derived dendritic cells (moDCs) and through in vivo immunization studies in a murine model, focusing on cellular immune responses. Results: GPs demonstrated stable physicochemical properties and efficient protein encapsulation, highlighting their suitability as vaccine delivery systems. They exhibited biocompatibility by not inducing cytotoxicity, hemolysis, or excessive ROS and NO production. In PBMCs, GPs stimulated cytokine secretion, suggesting their adjuvant potential. GPs were efficiently internalized by monocytes and led to specific chemokine secretion in stimulated moDCs. In a murine model, GPs induced distinctive cellular immune responses, including TNF-α and IFN-γ production and effector memory T cell activation. Conclusions: These findings emphasize GPs’ biocompatibility and immunomodulatory effects, highlighting their potential in immunotherapy and vaccine development, particularly for targeting infectious agents like hepatitis B virus. Full article

A simple and rapid precipitation process was successfully employed to prepare silver phosphate (SP, Ag₃PO₄). Two different phosphate sources: diammonium hydrogen phosphate ((NH₄)₂HPO₄) and dipotassium hydrogen phosphate (K₂HPO₄) were applied separately as the precursor, obtaining ((NH₄)₂HPO₄)⁻ and K₂HPO₄⁻ derived SP powders, named SP-A or SP-P, respectively. Fourier transform infrared (FTIR) spectra pointed out the vibrational characteristics of P–O and O–P–O interactions, confirming the presence of the PO43– functional group for SP. X-ray diffraction (XRD) patterns revealed that the SP crystallized in a cubic crystal structure. Whereas the field emission scanning electron microscope (FESEM) exposed spherical SP particles. The potentially antibacterial activity of SP-A and SP-P against bacterial Bacillus stratosphericus, yeast Meyerozyma guilliermondii, and fungal Phanerodontia chrysosporium was subsequently investigated. All studied microorganisms were recovered and isolated from the aquatic plant during the tissue culture process. The preliminary result of the antimicrobial test revealed that SP-A has higher antimicrobial activity than SP-P. The superior antimicrobial efficiency of SP-A compared to SP-P may be attributed to its purity and crystallite size, which provide a higher surface area and more active sites. In addition, the presence of potassium-related impurities in SP-P could have negatively affected its antimicrobial performance. These findings suggest that SP holds potential as an antimicrobial agent for maintaining sterility in tissue cultures, particularly in aquatic plant systems. The growth of both B. stratosphericus and M. guilliermondii was suppressed effectively at 30 ppm SP-A, whereas 10 ppm of SP-A can suppress P. chrysosporium development. This present work also highlights the potential of SP at very low concentrations (10–30 ppm) for utilization as an effective antimicrobial agent in tissue culture, compared to a commercial antimicrobial agent, viz., acetic acid, at the same concentration. Full article