Search Results (490)

Root-knot nematodes (RKNs) of the Meloidogyne genus impact various plants, including crops, fruits, and vegetables. Few chemical control options exist globally, and many nematicides are banned due to health and environmental risks. This study tested a new nematicidal agent, the symbiotic bacterium Xenorhabdus indica, which was molecularly identified (PV845100). Cell-free culture supernatants of Xenorhabdus spp. and their biogenic Ag-NPs were used in nematicidal assays. Meloidogyne incognita showed high mortality rates of 95.3%, 74.6%, and 72.6% after 72 h of treatment with the X. indica filtrate at three concentrations. At the same concentrations, biogenic Ag-NPs resulted in 82.0%, 90.0%, and 85.3% mortality rates, respectively. After 72 h, hatchability decreased by 53%, 74.6%, and 72.6% for the X. indica filtrate and 82.0%, 90.0%, and 85.3% for Ag-NPs. Quantitative real-time PCR (Q-PCR) revealed that Mi-Ache1 expression was lower in M. incognita second-stage juveniles (J2s) treated with the filtrate and Ag-NPs after 72 h compared to controls. Mi-Ache2 expression was also decreased, but only slightly. Furthermore, both the X. indica filtrate and biogenic Ag-NPs were safe in human lung (WI-38) and skin (HFB4) cell lines. These findings suggest that bacterial filtrates and their biogenic Ag-NPs could serve as cost-effective, environmentally friendly alternatives to commercial nematicides. Full article

As a typical representative of soft capping, primary vegetation capping has both protective and destructive effects on earthen city walls. Addressing its detrimental aspects constitutes the central challenge of this project. Because the integration of MICP technology with plants offered advantages, including soil solidification, erosion resistance, and resilience to dry–wet cycles and freeze–thaw cycles, the application of MICP technology to root–soil composites was proposed as a potential solution. Employing a combined approach of RF-RFE-CV modeling and microscopic imaging on laboratory samples from the Western City Wall of the Jinyang Ancient City in Taiyuan, Shanxi Province, China, key factors and characteristics in the mineralization process of Sporosarcina pasteurii were quantified and observed systematically to define the optimal pathway for enhancing urease activity and calcite yield. The conclusions were as follows. The urease activity of Sporosarcina pasteurii was primarily regulated by three key parameters with bacterial concentration, pH value, and the intensity of urease activity, which required stage-specific dynamic control throughout the growth cycle. Bacterial concentration consistently emerged as a high-importance feature across multiple time points, with peak effectiveness observed at 24 h (1.127). pH value remained a highly influential parameter across several time points, exhibiting maximum impact at around 8 h (1.566). With the intensity of urease activity, pH exerted a pronounced influence during the early cultivation stage, whereas inoculation volume gained increasing importance after 12 h. To achieve maximum urease activity, the use of CASO AGAR Medium 220 and the following optimized culture conditions was recommended: an activation culture time of 27 h, an inoculation age of 16 h, an inoculation volume of 1%, a culture temperature of 32 °C, an initial pH of 8, and an oscillation speed of 170 r/min. Furthermore, to maximize the yield of CaCO₃ in output and the yield of calcite in CaCO₃, the following conditions and procedures were recommended: a ratio of urea concentration to Ca²⁺ concentration of 1 M:1.3 M, using the premix method of Sporosarcina pasteurii, quiescent reaction, undisturbed filtration, and drying at room-temperature in the shade environment. Full article

Agro-industrial residues rich in carbohydrates represent low-cost and sustainable feedstock for enzyme production. This study demonstrates that green Arabica coffee press cake, a mannan-rich coproduct of oil extraction, is an efficient carbon source for Aspergillus niger (CFAM 1234) cultivation and for inducing mannanase production. Furthermore, the enzymes obtained were tested for mannose recovery in the enzymatic hydrolysis of healthy and defective coffee beans to investigate their hydrolytic potential. Mannanase production was investigated using various carbon sources—including ground coffee beans; coffee press cake; different particle sizes of coffee press cake; aqueous coffee cake extract (prepared at 30 g·L⁻¹ under constant stirring (300 rpm) at 80 °C for 2 h, followed by filtration.); and a commercial galactomannan, locust bean gum (LBG). CNHSO analysis was performed in the best carbon source (coffee press cake) and LBG. Statistical optimization (Plackett–Burman and Central Composite Rotatable Design) simplified the culture medium composition to coffee press cake (48.78 g·L⁻¹), yeast extract (4 g·L⁻¹), and potassium phosphate (0.25 g·L⁻¹, pH 5.5) and increased mannanases productivity to 22.4 ± 0.6 U·mL⁻¹ within only 3 days (a 42.9% improvement compared to non-optimized conditions, which were 30 g·L⁻¹, carbon source, 4 g·L⁻¹ yeast extract, 1 g·L⁻¹ Al₂O₃, 0.5 g·L⁻¹ potassium phosphate buffer (pH 5.5), 0.5 g·L⁻¹ of MgSO₄·7H₂O, and 0.05 g·L⁻¹ of CaCl₂·2H₂O, which resulted in a maximum of ~20 U·mL⁻¹ in 7 days). The crude extract also exhibited β-mannosidase activity (1.39 ± 0.06 U·mL⁻¹). When applied to the hydrolysis of untreated healthy and defective coffee beans, the enzyme preparation enabled ~25% mannose recovery (considering the value obtained through acid hydrolysis as 100%), highlighting its potential as a mannose resource. The results demonstrate that coproducts from the coffee production chain can be used as an efficient carbon source (coffee cake) for mannanase production, as well as sugar recovery (defective coffee beans), offering an integrated strategy to strengthen the circular bioeconomy and generate carbohydrates with potential industrial and nutritional applications. Full article

Biological control is an effective and environmentally friendly strategy for managing plant diseases. In this study, a broad-spectrum antagonistic bacterium, designated strain AN6, was isolated from rice plants and exhibited potent inhibitory activity against a variety of phytopathogens. In Oxford cup assays, AN6 suppressed the growth of Xanthomonas oryzae pv. oryzae (Xoo) by 73.60%, and its cell-free culture filtrate caused pronounced morphological deformation in the bacterial cells. Further in vitro assays, including dual-culture assays, volatile organic compound (VOC) assays, and cell-free supernatant (CFS) assays, demonstrated that AN6 also exerted strong antifungal effects against several pathogenic fungi. In addition, the strain was found to produce proteases and siderophores, which may contribute to its antagonistic capabilities. Taxonomic identification based on morphological traits, 16S rRNA and gyrA gene sequencing, average nucleotide identity (ANI), in silico DNA–DNA hybridization (isDDH), and phylogenetic analysis classified strain AN6 as Bacillus velezensis. Whole-genome sequencing revealed that AN6 harbors a 3,929,788 bp genome comprising 4025 protein-coding genes with a GC content of 46.50%. Thirteen biosynthetic gene clusters (BGCs) associated with the production of secondary metabolites—such as nonribosomal peptides, polyketides, and dipeptide antibiotics—were identified. The pot experiment further validated the biocontrol potential of AN6, achieving an 80.49% reduction in rice bacterial blight caused by Xanthomonas oryzae pv. oryzae. Collectively, these results indicate that B. velezensis AN6 is a promising candidate for development as a highly effective biocontrol agent for the integrated management of diverse plant diseases. Full article

This study aimed to determine the culture conditions and some anatomic features for using white worms (Enchytraeus sp.) as live feed in aquaculture. In the first experiment (Culture 1), coconut peat was used as the culture media and it was determined that, under approximately 21 °C, the population of the white worm species increased from 10 adult individuals to a total of 424 individuals (including both adults and juveniles) over a 70-day period. Unlike the first experimental stage (Culture 1), the second stage (Culture 2) was structured into three groups. In the second experiment (Culture 2), coconut peat was used as the culture media in the first (G1) and second experimental groups (G2), and zeolite with coconut peat in the third group (G3). The first (G1) and third experimental (G3) groups were fed a control diet, while the second group (G2) was fed a diet with 10% zeolite added to the control diet. In the 60-day study conducted during the culture-2 phase, the worm population, which initially consisted of 10 individuals, reached its highest values in Group 3. The use of zeolite as a substrate and feed additive for white worm culture was investigated for the first time, and original data were obtained that will contribute to filling the gap in the literature. Furthermore, the aquarium filtration sponge used in addition to the culture soil was determined to be a suitable substrate material for collecting cocoon data. The findings of this study provide new supporting data that will fill gaps in the literature regarding materials and methods that contribute positively to white worm culture conditions. Full article

Rice sheath blight caused by Rhizoctonia solani is a devastating global rice disease. This study aimed to isolate biocontrol bacteria from the medicinal plant Bletilla striata for managing the disease. Strain A1 demonstrated the strongest antagonistic activity, with a 91.92% inhibition rate against R. solani in vitro. It also exhibited a broad antifungal spectrum against ten plant pathogenic fungi. Morphological and molecular (16S rRNA and recA genes) analysis identified strain A1 as Pseudomonas koreensis. In detached leaf assays, lesion length was significantly reduced. Pot and field trials showed control efficacies of 65.54% and 72.53%, respectively, comparable to the chemical agent Jinggangmycin. Strain A1 secreted extracellular enzymes (protease, β-1,3-glucanase), siderophores, and auxin (IAA), and possessed phosphate-solubilizing and nitrogen-fixing capabilities. The strain significantly enhanced the activities of key defense enzymes (POD, PAL, PPO, CAT, SOD) in rice. Furthermore, both its sterile culture filtrate and the corresponding crude ethyl acetate extract exhibited strong, direct suppression of R. solani growth. LC-MS analysis identified potential antifungal compounds, including Pseudomonic Acid, Artemisinin, and Tetradecane, in the extract. In conclusion, P. koreensis A1 is a promising biocontrol and plant growth-promoting candidate for sustainable management of rice sheath blight. Full article

Recurrent Clostridioides difficile infection (rCDI) remains a major therapeutic challenge. Although faecal microbiota transplantation (FMT) is highly effective and thought to restore microbial composition and metabolic function, the mechanisms underlying its success are not fully understood. In particular, the contribution of non-bacterial components such as soluble metabolites remains unclear. Therefore, further investigation is needed to identify the mechanistic drivers of FMT efficacy and clarify how non-bacterial factors contribute to therapeutic outcomes. Here, we applied untargeted three-dimensional Orbitrap secondary ion mass spectrometry (3D OrbiSIMS) to profile faecal metabolic reprogramming in rCDI patients pre- and post-FMT, alongside C. difficile cultures exposed to sterile faecal filtrates. FMT induced extensive metabolic shifts, restoring glyoxylate/dicarboxylate and glycerophosphoinositol pathways and normalising disrupted bile acid and amino acid profiles. Faecal filtrate exposure caused strain-specific metabolic disruption in C. difficile, depleting proline, fumarate and succinate while enriching tryptophan. While multiple metabolite classes were profiled, the most significant functional changes were observed in lipids. Lipidomics identified >3.8-fold enrichment of phosphatidylinositol (PI) species, which localised to bacterial membranes and conferred cytoprotection against C. difficile toxins and other epithelial insults. Spatial metabolomics imaging revealed, for the first time, metabolite compartmentalisation within C. difficile, with proline and succinate broadly distributed across the cell surface and fumarate confined to distinct microdomains, highlighting functional heterogeneity in pathogen metabolism. Collectively, these findings demonstrate that soluble metabolites within faecal filtrates mediate pathogen suppression and epithelial barrier protection, establishing metabolite-driven mechanisms underlying FMT efficacy and identifying PI lipids as candidate post-biotic therapeutics for rCDI. Full article

This study evaluated the feasibility of culturing Seriola lalandi in a low-cost recirculating aquaculture system (RAS) in an arid region of northern Chile, aiming to establish strategies for broodstock farming and diversify national aquaculture. The system was designed as a low-cost recirculating aquaculture system (RAS) built with locally available materials, such as galvanized corrugated steel panels and flexible plastic liners, instead of specialized aquaculture tanks. Its modular configuration, based on gravity-fed filtration using sedimentation, sand, and disc filters, allows efficient water reuse with minimal energy consumption and a daily water turnover of 12 times the total volume. This design significantly reduced construction and operational costs, making it a feasible option for aquaculture development in arid regions with limited water resources. Over an 8-month period, 46 S. lalandi individuals were used, and the results showed successful physiological adaptation of the specimens to confinement, as evidenced by low mortality, progressive acceptance of formulated feed, and sustained growth. Individual weights progressively increased, with averages ranging from 675 to 1435 g, and the specific growth rate (SGR) fluctuated between 0.14 and 0.43% per day. Fulton’s condition factor (K) remained in an adequate range between 2.4 and 2.8, suggesting good physical condition of the sampled individuals. Water quality within the RAS system was maintained within acceptable parameters, although a strong negative correlation between temperature and dissolved oxygen was recorded (Spearman coefficient = −0.71, p < 0.001), highlighting the importance of monitoring these factors in warm environments. The lack of adequate protocols for the adaptation of marine species in arid areas, such as northern Chile, has limited aquaculture development in these regions. This study addresses this problem by assessing the feasibility of a low-cost recirculating system (RAS) for the cultivation of S. lalandi under conditions of water scarcity, with the aim of diversifying the national aquaculture in arid zones. Full article

Galactooligosaccharides (GOS) typically consist of 2-8 D-galactose units linked together, terminating in a D-glucose unit. GOS are commonly used in dairy products, infant formulas, and functional foods. GOS offer beneficial properties for food processing, such as low caloric value, mild clean taste, and excellent solubility in water. Additionally, GOS function as non-digestible prebiotics, supporting microbiota balance and offering benefits such as promoting infant health, immune modulation, laxative effects, and potential metabolic advantages. β-galactosidase plays a key role in GOS production, catalyzing both hydrolysis and transglycosylation reactions. In this study, a putative GH2 family β-galactosidase from Kosakonia oryzendophytica (Koor β-gal) was identified. The enzyme exhibited optimal activity at pH 7.0 and 45–50 °C with the addition of 1 mM Mg²⁺, showing a specific activity of approximately 288.6 U/mg towards o-nitrophenyl-β-D-galactopyranoside (ONPG). After optimizing the reaction conditions, Koor β-gal successfully produced 124.7 g/L of GOS from 300 g/L D-lactose, achieving a GOS yield of 41.6%. LC-MS analysis revealed that the primary products consisted of GOS with degrees of polymerization (DP) ranging from 2 to 4. Additionally, Koor β-gal was heterologously expressed in Bacillus subtilis following comprehensive optimization of the promoter and 5′-UTR, resulting in an enzyme activity in culture filtrate of 106.2 U/mL after 60 h. Full article

Methane is a potent greenhouse gas that requires its emissions to be mitigated. A significant source for methane emissions is in the form of the biogas that is produced from anaerobic digestion in wastewater reclamation and landfill facilities. Biogas has a high valorization potential in the form of its bioconversion into ectoines, an active ingredient in skin care products, by halotolerant alkaliphilic methanotrophs. Cultures of Methylotuvimicrobium alcaliphilum 20Z were grown in bench scale stirred-tank reactors to determine factors to improve methane uptake and removal. Tangential flow filtration was also implemented for a bio-milking method to recover ectoine from culture media. Methane uptake and reactor productivity increased, with a temperature of 28 °C compared with 21 °C. Decreasing the methane gas bubble diameter by decreasing the sparger pore size from 1 mm to 0.5 µm significantly improved methane removal and reactor productivity by increasing mass transfer. Premixing methane and air before sparging into the reactor saw a higher removal of methane, while sparging methane and air separately created an increase in reactor productivity. Maximum methane removal efficiency was observed to be 70.56% ± 0.54 which translated to a CH₄-EC of 93.82 ± 3.36 g CH₄ m⁻³ h⁻¹. Maximum ectoine yields was observed to be 0.579 mg ectoine L⁻¹ h⁻¹. Full article

Rapid urban expansion and increasing population density intensify the loss of open spaces, exacerbate flooding frequency and runoff pollution, increase the urban heat island effect, and deteriorate ecological resilience and human well-being. This study presents Gazelle Valley Park (GVP) in Jerusalem (Israel), a unique large-scale ecohydrological infrastructure within a dense Mediterranean city. GVP was established in 2015 following a public-led campaign and comprises a multifunctional nature-based solution designed to collect and circulate stormwater through a series of vegetated ponds, enhancing filtration, aeration, and pollutant removal, while sustaining a wetland ecosystem. Its design follows international ecological standards and embodies the principle “from nuisance to resource”, transforming urban runoff into an asset that supports rich biodiversity while offering recreational, cultural, and educational activities. During the dry summer, reclaimed wastewater is introduced in order to support a perennial aquatic habitat, which introduces various challenges due to increased salinity, oxygen demand, and contaminants. Hydrometric and geochemical monitoring demonstrates strong correlations between rainfall and runoff and point at the role of sedimentation and vegetation in reducing pollutant loads. The park benefits from its holistic operation, where hydrology, ecology, education, and public engagement are integrated, thus making the whole greater than the sum of its parts. Full article

We previously demonstrated that in BAP1-proficient pleural mesothelioma cells, CDKN2A is critical for mediating the response to selective EZH2 inhibition and highlighted a complex interplay between epigenetic regulation and the tumor immune microenvironment. In this study, we employed a quantitative proteomic mass spectrometry approach to assess alterations in protein expression following EZH2 inhibition in BAP1- and CDKN2A-proficient mesothelioma cells cultured as spheroids. Additionally, we analyzed extracellular vesicles (EVs), which were isolated through tangential flow filtration. Flow cytometric analysis and co-culture systems were used to characterize the effects of EVs on neutrophils. Upon EZH2 inhibition, we demonstrated RAB27b and CD63 upregulation and increased release of extracellular vesicles. We found that a brief exposure to EVs derived from EZH2 inhibitor-treated cells skewed naïve neutrophils toward a pro-tumor phenotype characterized by high levels of PD-L1 and MSLN (Mesothelin) expression on the surface. These EV-elicited neutrophils suppressed T cell proliferation while enhancing tumor cell growth. Moreover, we observed changes in the EV cargo derived from EZH2 inhibitor-treated spheroids. Our findings highlight the significant role of EVs in creating an immunosuppressive microenvironment, and underscore the urgent need for further investigation into the regulation of neutrophil biology and function in the PM. Full article

The Symbiotic Culture of Bacteria and Yeast (SCOBY) is a cellulose-based biofilm resulting from the fermentation of sweetened tea by a microbial consortium of acetic acid bacteria and yeasts. This study applies the Methodi Ordinatio technique to systematically identify, rank, and analyze the most relevant scientific publications on the applications of SCOBY. A comprehensive search in SCOPUS and Web of Science yielded 179 articles, after manual filtration. The InOrdinatio index, which combines citation count, publication year, and journal impact factor, was used for ranking to select a representative sample of the most important contributions (117 articles). The highest-ranked article scored 128.9, and the lowest 42.6. China led in scientific output (14.01%), followed by India (11.46%), the UK and USA (5.10% each), and Brazil (4.46%). The International Journal of Biological Macromolecules was the most frequently used journal for publications in this field. “Bacterial cellulose” was the most cited keyword (61 times), followed by “kombucha” (41) and “fermentation” (29). A consistent rise in publications has been observed over the past five years. Four main application areas were identified: bacterial cellulose (BC) (38%), biosustainable materials (28%), biomedical (17%), and food-related uses (17%). Most of the studies related to BC production (52%) searched for alternative substrates, and 18% focused on the isolation and identification of the most productive microorganisms within SCOBY. For biomedical applications, a unifying theme is the development of SCOBY-based materials with intrinsic antibacterial properties. These findings emphasize SCOBY’s emerging role in sustainable innovation and circular economic frameworks. Full article

Background: Recent advances in antibody discovery technologies, especially progress in de novo synthesis through machine learning, have imposed a significant production challenge for the generation of a large diversity of antibodies against nearly any target of interest. There is a demand for the rapid production of dozens of purified antibodies in 10-milligram quantities sufficient for functional screening and molecular assessment studies. Objectives: To meet this requirement, a semi-automated production methodology and workflow was developed to bridge the miniaturized high-throughput screenings (HTSs) and the conventional custom-scale workflow by taking advantage of four new technology applications. Methods: First, it exploited a novel, simple, high-titer transient expression system, “CHO4Tx^®”, which could achieve high yields in the range of 200 mg/L and above, across a variety of antibody constructs, including challenging targets. The consistently high yields from this transient CHO platform enabled the delivery of ~20 mg of crude material per 100 mL scale flask production with a throughput capacity of nineteen constructs in a single run. Secondly, we established a magnetic ProA bead in-culture antibody-capturing process, which significantly shortened the production timeline by eliminating the steps of cell centrifugation, filtration, and medium column loading. Third, we utilized the GenScript AmMag™ SA Plus semi-automation, which could handle magnetic ProA bead elution for 12 constructs within less than 1 h. Lastly, we transformed the AKTA Pure^TM system into an automated buffer exchange purification system with a capacity of processing 19 samples in a single run. Results and Conclusions: This new production platform was proven to be robust and could be applied for the routine production of antibodies of sufficient quality and quantity in support of cell-based assays and biophysical characterization. Full article

Glottic insufficiency, often caused by laryngeal nerve injury, impairs voice quality and breathing. Current treatments, such as hyaluronic acid injection, require frequent reapplication every 3–6 months. This study aimed to investigate the therapeutic potential of small extracellular vesicles (sEVs) derived from Wharton’s Jelly mesenchymal stem cells (WJMSCs) incorporated into genipin-crosslinked gelatin hydrogels (GCGHs) for promoting vocal fold fibroblast (VFFs) regeneration in vitro. WJMSCs were isolated from umbilical cords, expanded to passage 4, and used for sEV isolation via tangential flow filtration (TFF). The sEVs (585.89 ± 298.93 µg/mL) were characterized using bicinchoninic acid assay (BCA), nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and Western blot. Seven concentrations of sEVs were tested on VFFs to evaluate cytotoxicity and proliferation, identifying 75 µg/mL as the optimal dose. GCGHs were then combined with WJMSCs and sEVs and evaluated for physicochemical properties, degradation, biocompatibility, and immune response. The hydrogels were injectable within 20 min and degraded in approximately 42 ± 0.72 days. The optimal sEV concentration significantly enhanced VFFs proliferation (166.59% ± 28.11) and cell viability (86.16% ± 8.55, p < 0.05). GCGH-MSCs showed the highest VFFs viability (82.04% ± 10.51) and matrix contraction (85.98% ± 1.25) compared to other groups. All hydrogel variants demonstrated minimal immune response when co-cultured with peripheral blood mononuclear cells (PBMCs). GCGH is a promising scaffold for delivering WJMSCs and sEVs to support VFF regeneration, with demonstrated biocompatibility and regenerative potential. Further in vivo studies are warranted to validate these findings. Full article