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18 pages, 7363 KiB  
Article
Agronomic Evaluation of Compost Formulations Based on Mining Tailings and Microbial Mats from Geothermal Sources
by María Jesús Puy-Alquiza, Miren Yosune Miranda Puy, Raúl Miranda-Avilés, Pooja Vinod Kshirsagar and Cristina Daniela Moncada Sanchez
Recycling 2025, 10(4), 156; https://doi.org/10.3390/recycling10040156 - 5 Aug 2025
Viewed by 86
Abstract
This study, conducted in Mexico, evaluates the agricultural potential of three compost formulations BFS1, BFS2, and BFS3 produced from mining tailings and thermophilic microbial mats and collected from geothermal environments. The physicochemical characterization included pH, electrical conductivity (EC), macronutrients (N, P, K, Ca, [...] Read more.
This study, conducted in Mexico, evaluates the agricultural potential of three compost formulations BFS1, BFS2, and BFS3 produced from mining tailings and thermophilic microbial mats and collected from geothermal environments. The physicochemical characterization included pH, electrical conductivity (EC), macronutrients (N, P, K, Ca, Mg, and S), micronutrients (Fe, Zn, B, Cu, Mn, Mo, and Ni), organic matter (OM), and the carbon-to-nitrogen (C/N) ratio. All composts exhibited neutral pH values (7.38–7.52), high OM content (38.5–48.4%), and optimal C/N ratios (10.5–13.9), indicating maturity and chemical stability. Nitrogen ranged from 19 to 21 kg·t−1, while potassium and calcium were present in concentrations beneficial for crop development. However, EC values (3.43–3.66 dS/m) and boron levels (>160 ppm) were moderately high, requiring caution in saline soils or with boron-sensitive crops. A semi-quantitative Compost Quality Index (CQI) ranked BFS3 highest due to elevated OM and potassium content, followed by BFS1. BFS2, while rich in nitrogen, scored lower due to excessive boron. One-way ANOVA revealed no significant difference in nitrogen (p > 0.05), but it did reveal significant differences in potassium (p < 0.01) and boron (p < 0.001) among formulations. These results confirm the potential of mining tailings—microbial mat composts are low-cost, nutrient-rich biofertilizers. They are suitable for field crops or as components in nursery substrates, particularly when EC and boron are managed through dilution. This study promotes the circular reuse of geothermal and industrial residues and contributes to sustainable soil restoration practices in mining-affected regions through innovative composting strategies. Full article
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23 pages, 2657 KiB  
Article
Enrichment Cultures of Extreme Acidophiles with Biotechnological Potential
by Khussain Valiyev, Aliya Yskak, Elena Latyuk, Alena Artykova, Rakhimbayev Berik, Vadim Chashkov and Aleksandr Bulaev
Mining 2025, 5(3), 49; https://doi.org/10.3390/mining5030049 - 1 Aug 2025
Viewed by 101
Abstract
The purpose of this work was to obtain specialized enrichment cultures from an original extreme acidophilic consortium of extremely acidophilic microorganisms and to study their microbial community composition and biotechnological potential. At temperatures of 25, 35, 40 and 50 °C, distinct enrichments of [...] Read more.
The purpose of this work was to obtain specialized enrichment cultures from an original extreme acidophilic consortium of extremely acidophilic microorganisms and to study their microbial community composition and biotechnological potential. At temperatures of 25, 35, 40 and 50 °C, distinct enrichments of extremely acidophilic microorganisms used in the processes of bioleaching sulfide ores were obtained using nutrient media containing ferrous sulfate, elemental sulfur and a copper sulfide concentrate as nutrient inorganic substrates, with and without the addition of 0.02% yeast extract. The microbial community composition was studied using the sequencing of the V3–V4 hypervariable region of the 16S rRNA genes. The different growth conditions led to changes in the microbial composition and relative abundance of mesophilic and moderately thermophilic, strict autotrophic and mixotrophic microorganisms in members of the genera Acidithiobacillus, Sulfobacillus, Leptospirillum, Acidibacillus, Ferroplasma and Cuniculiplasma. The dynamics of the oxidation of ferrous iron, sulfur, and sulfide minerals (pyrite and chalcopyrite) by the enrichments was also studied in the temperature range of 25 to 50 °C. The study of enrichment cultures using the molecular biological method using the metabarcoding method of variable V3–24 V4 fragments of 16S rRNA genes showed that enrichment cultures obtained under different conditions differed in composition, which can be explained by differences in the physiological properties of the identified microorganisms. Regarding the dynamics of the oxidation of ferrous ions, sulfur, and sulfide minerals (pyrite and chalcopyrite), each enrichment culture was studied at a temperature range of 25 to 50 °C and indicated that all obtained enrichments were capable of oxidizing ferrous iron, sulfur and minerals at different rates. The obtained enrichment cultures may be used in further work to increase bioleaching by using the suitable inoculum for the temperature and process conditions. Full article
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14 pages, 4194 KiB  
Article
Crystal Structure of Anthranilate Phosphoribosyltransferase from Methanocaldococcus jannaschii
by Jung-Min Choi
Crystals 2025, 15(8), 702; https://doi.org/10.3390/cryst15080702 - 31 Jul 2025
Viewed by 172
Abstract
Tryptophan is synthesized in microorganisms via a five-step enzymatic pathway originating from chorismate, which is a product of the shikimate pathway. As a biosynthetic precursor to a wide range of high-value compounds such as indole-3-acetic acid, indigo, indirubin, and violacein, this pathway has [...] Read more.
Tryptophan is synthesized in microorganisms via a five-step enzymatic pathway originating from chorismate, which is a product of the shikimate pathway. As a biosynthetic precursor to a wide range of high-value compounds such as indole-3-acetic acid, indigo, indirubin, and violacein, this pathway has been a central target for metabolic engineering to enhance microbial production. Anthranilate phosphoribosyltransferase (AnPRT) catalyzes the second step of the pathway by transferring a phosphoribosyl group from PRPP to anthranilate, forming phosphoribosyl anthranilate (PRA). AnPRT, the sole member of class IV phosphoribosyltransferases, adopts a unique fold and functions as a homodimer. While the structural basis of AnPRT activity has been elucidated in several organisms, thermostable variants remain underexplored despite their relevance for high-temperature bioprocessing. In this study, the crystal structure of AnPRT from the thermophilic archaeon Methanocaldococcus jannaschii (MjAnPRT) was determined at a 2.16 Å resolution. The enzyme exhibits a conserved dimeric architecture and key catalytic motifs. Comparative structural analysis with mesophilic and hyper thermophilic homologs revealed that MjAnPRT possesses enhanced local stability in catalytically important regions and strengthened inter-subunit interactions. These features likely contribute to its thermostability and provide a valuable framework for the rational design of robust AnPRTs for industrial and synthetic biology applications. Full article
(This article belongs to the Special Issue Crystallography of Enzymes)
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29 pages, 3259 KiB  
Review
The Role of the Environment (Water, Air, Soil) in the Emergence and Dissemination of Antimicrobial Resistance: A One Health Perspective
by Asma Sassi, Nosiba S. Basher, Hassina Kirat, Sameh Meradji, Nasir Adam Ibrahim, Takfarinas Idres and Abdelaziz Touati
Antibiotics 2025, 14(8), 764; https://doi.org/10.3390/antibiotics14080764 - 29 Jul 2025
Viewed by 439
Abstract
Antimicrobial resistance (AMR) has emerged as a planetary health emergency, driven not only by the clinical misuse of antibiotics but also by diverse environmental dissemination pathways. This review critically examines the role of environmental compartments—water, soil, and air—as dynamic reservoirs and transmission routes [...] Read more.
Antimicrobial resistance (AMR) has emerged as a planetary health emergency, driven not only by the clinical misuse of antibiotics but also by diverse environmental dissemination pathways. This review critically examines the role of environmental compartments—water, soil, and air—as dynamic reservoirs and transmission routes for antibiotic-resistant bacteria (ARB) and resistance genes (ARGs). Recent metagenomic, epidemiological, and mechanistic evidence demonstrates that anthropogenic pressures—including pharmaceutical effluents, agricultural runoff, untreated sewage, and airborne emissions—amplify resistance evolution and interspecies gene transfer via horizontal gene transfer mechanisms, biofilms, and mobile genetic elements. Importantly, it is not only highly polluted rivers such as the Ganges that contribute to the spread of AMR; even low concentrations of antibiotics and their metabolites, formed during or after treatment, can significantly promote the selection and dissemination of resistance. Environmental hotspots such as European agricultural soils and airborne particulate zones near wastewater treatment plants further illustrate the complexity and global scope of pollution-driven AMR. The synergistic roles of co-selective agents, including heavy metals, disinfectants, and microplastics, are highlighted for their impact in exacerbating resistance gene propagation across ecological and geographical boundaries. The efficacy and limitations of current mitigation strategies, including advanced wastewater treatments, thermophilic composting, biosensor-based surveillance, and emerging regulatory frameworks, are evaluated. By integrating a One Health perspective, this review underscores the imperative of including environmental considerations in global AMR containment policies and proposes a multidisciplinary roadmap to mitigate resistance spread across interconnected human, animal, and environmental domains. Full article
(This article belongs to the Special Issue The Spread of Antibiotic Resistance in Natural Environments)
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22 pages, 2743 KiB  
Article
Effects of the Application of Different Types of Vermicompost Produced from Wine Industry Waste on the Vegetative and Productive Development of Grapevine in Two Irrigation Conditions
by Fernando Sánchez-Suárez, María del Valle Palenzuela, Cristina Campos-Vazquez, Inés M. Santos-Dueñas, Víctor Manuel Ramos-Muñoz, Antonio Rosal and Rafael Andrés Peinado
Agriculture 2025, 15(15), 1604; https://doi.org/10.3390/agriculture15151604 - 25 Jul 2025
Viewed by 324
Abstract
This study evaluates the agronomic potential of two types of vermicompost—one produced solely from wine industry residues (WIR) and one incorporating sewage sludge (WIR + SS)—under rainfed and deficit irrigation conditions in Mediterranean vineyards. The vermicompost was obtained through a two-phase process involving [...] Read more.
This study evaluates the agronomic potential of two types of vermicompost—one produced solely from wine industry residues (WIR) and one incorporating sewage sludge (WIR + SS)—under rainfed and deficit irrigation conditions in Mediterranean vineyards. The vermicompost was obtained through a two-phase process involving initial thermophilic pre-composting, followed by vermicomposting using Eisenia fetida for 90 days. The conditions were optimized to ensure aerobic decomposition and maintain proper moisture levels (70–85%) and temperature control. This resulted in end products that met the legal standards required for agricultural use. However, population dynamics revealed significantly higher worm reproduction and biomass in the WIR treatment, suggesting superior substrate quality. When applied to grapevines, WIR vermicompost increased soil organic matter, nitrogen availability, and overall fertility. Under rainfed conditions, it improved vegetative growth, yield, and must quality, with increases in yeast assimilable nitrogen (YAN), sugar content, and amino acid levels comparable to those achieved using chemical fertilizers, as opposed to the no-fertilizer trial. Foliar analyses at veraison revealed stronger nutrient uptake, particularly of nitrogen and potassium, which was correlated with improved oenological parameters compared to the no-fertilizer trial. In contrast, WIR + SS compost was less favorable due to lower worm activity and elevated trace elements, despite remaining within legal limits. These results support the use of vermicompost derived solely from wine residues as a sustainable alternative to chemical fertilizers, in line with the goals of the circular economy in viticulture. Full article
(This article belongs to the Special Issue Vermicompost in Sustainable Crop Production—2nd Edition)
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15 pages, 3942 KiB  
Article
Quantitative Evaluation of Endogenous Reference Genes for RT-qPCR and ddPCR Gene Expression Under Polyextreme Conditions Using Anaerobic Halophilic Alkalithermophile Natranaerobius thermophilus
by Xinyi Tao, Qinghua Xing, Yingjie Zhang, Belsti Atnkut, Haozhuo Wei, Silva Ramirez, Xinwei Mao and Baisuo Zhao
Microorganisms 2025, 13(8), 1721; https://doi.org/10.3390/microorganisms13081721 - 23 Jul 2025
Viewed by 290
Abstract
Accurate gene expression quantification using reverse transcription quantitative PCR (RT-qPCR) requires stable reference genes (RGs) for reliable normalization. However, few studies have systematically identified RGs suitable for simultaneous high salt, alkaline, and high-temperature conditions. This study addresses this gap by evaluating the stability [...] Read more.
Accurate gene expression quantification using reverse transcription quantitative PCR (RT-qPCR) requires stable reference genes (RGs) for reliable normalization. However, few studies have systematically identified RGs suitable for simultaneous high salt, alkaline, and high-temperature conditions. This study addresses this gap by evaluating the stability of eight candidate RGs in the anaerobic halophilic alkalithermophile Natranaerobius thermophilus JW/NM-WN-LFT under combined salt, alkali, and thermal stresses. The stability of these candidate RGs was assessed using five statistical algorithms: Delta CT, geNorm, NormFinder, BestKeeper, and RefFinder. Results indicated that recA exhibited the highest expression stability across all tested conditions and proved adequate as a single RG for normalization in both RT-qPCR and droplet digital PCR (ddPCR) assays. Furthermore, recA alone or combined with other RGs (sigA, rsmH) effectively normalized the expression of seven stress-response genes (proX, opuAC, mnhE, nhaC, trkH, ducA, and pimT). This work represents the first systematic validation of RGs under polyextreme stress conditions, providing essential guidelines for future gene expression studies in extreme environments and aiding research on microbial adaptation mechanisms in halophilic, alkaliphilic, and thermophilic microorganisms. Full article
(This article belongs to the Section Environmental Microbiology)
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13 pages, 785 KiB  
Article
Selective Lactic Acid Production via Thermophilic Anaerobic Fermentation
by Claudia Chao-Reyes, Rudolphus Antonius Timmers, Ahmed Mahdy, Silvia Greses and Cristina González-Fernández
Water 2025, 17(15), 2183; https://doi.org/10.3390/w17152183 - 22 Jul 2025
Viewed by 294
Abstract
The combined effect of temperature-adapted inocula and anaerobic fermentation (AF) settings (pH 5.1 and 50 °C) were assessed to produce short-chain carboxylates (SCCs). In this study, the AF of carrot pulp was investigated using inocula adapted at different temperatures (25, 35, and 55 [...] Read more.
The combined effect of temperature-adapted inocula and anaerobic fermentation (AF) settings (pH 5.1 and 50 °C) were assessed to produce short-chain carboxylates (SCCs). In this study, the AF of carrot pulp was investigated using inocula adapted at different temperatures (25, 35, and 55 °C) with the aim of shifting the microbiome activity from biogas to SCC production. The highest SCC content (17.2 g COD L−1), and bioconversion (26.1%) and acidification efficiency (56.3%) were achieved with 35 °C-adapted inoculum. Lactic acid production prevailed in all reactors, demonstrating a high selectivity in SCC production. Both the microbial richness and diversity sharply diminished in the 35 °C and 55 °C operated reactors, with Firmicutes phylum identified as key players of the lactic acid production in AF. The results demonstrated that the operating temperature played a key role in shaping the microbial structure of inocula, leading to different process performances and highlighting thermophilic AF as a feasible process to produce lactic acid. Full article
(This article belongs to the Special Issue Innovations in Anaerobic Digestion Technology)
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21 pages, 8441 KiB  
Article
Upper Pleistocene Marine Levels of the Es Copinar–Es Estufadors (Formentera, Balearic Islands, West Mediterranean)
by Laura del Valle, Guillem X. Pons and Joan J. Fornós
Quaternary 2025, 8(3), 38; https://doi.org/10.3390/quat8030038 - 21 Jul 2025
Viewed by 451
Abstract
Late Pleistocene coastal deposits on the southeastern coast of Formentera (Es Ram–Es Estufadors) provide a high-resolution record of sea-level and climatic fluctuations associated with Marine Isotope Stage (MIS) 5. Three distinct beach levels (Sef-1, Sef-2, Sef-3) were identified, corresponding to substages MIS 5e, [...] Read more.
Late Pleistocene coastal deposits on the southeastern coast of Formentera (Es Ram–Es Estufadors) provide a high-resolution record of sea-level and climatic fluctuations associated with Marine Isotope Stage (MIS) 5. Three distinct beach levels (Sef-1, Sef-2, Sef-3) were identified, corresponding to substages MIS 5e, 5c, and possibly 5a, based on sedimentological features, fossil assemblages, and Optically Stimulated Luminescence (OSL) dating. The oldest beach level (Sef-1) is attributed to MIS 5e (ca. 128–116 ka) and is characterised by the widespread presence of thermophilic Senegalese fauna—including Thetystrombus latus, Conus ermineus, and Linatella caudata—which mark the onset of this interglacial phase and are associated with two peaks in relative sea-level highstand. A subsequent cooling event during MIS 5d is recorded by the development of thin palaeosols and the disappearance of these warm-water taxa. The second beach level (Sef-2) reflects renewed sea-level rise and warmer conditions during MIS 5c, with abundant macrofauna and red algae. The transition to MIS 5b (~97 ka) is marked by a significant sea-level drop (down to –60 m), cooler climate, and enhanced colluvial sedimentation linked to increased runoff and erosion. In total, 54 macrofaunal species were identified—16 from Sef-1 and 46 from Sef-2—highlighting ecological shifts across substages. These results improve our understanding of coastal response to sea-level oscillations and paleoenvironmental dynamics in the western Mediterranean during the Late Pleistocene. Full article
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20 pages, 3201 KiB  
Article
Effect of Screw Configuration on the Recyclability of Natural Fiber-Based Composites
by Vlasta Chyzna, Steven Rowe, James Finnerty, Trevor Howard, Christopher Doran, Shane Connolly, Noel Gately, Alexandre Portela, Alan Murphy, Declan M. Devine and Declan Mary Colbert
Fibers 2025, 13(7), 98; https://doi.org/10.3390/fib13070098 - 18 Jul 2025
Viewed by 401
Abstract
The burgeoning crisis of plastic waste accumulation necessitates innovative approaches towards sustainable packaging solutions. Polylactic acid (PLA), a leading biopolymer, emerges as a promising candidate in this realm, especially for environmentally friendly packaging. PLA is renowned for its compostable properties, offering a strategic [...] Read more.
The burgeoning crisis of plastic waste accumulation necessitates innovative approaches towards sustainable packaging solutions. Polylactic acid (PLA), a leading biopolymer, emerges as a promising candidate in this realm, especially for environmentally friendly packaging. PLA is renowned for its compostable properties, offering a strategic avenue to mitigate plastic waste. However, its dependency on specific industrial composting conditions, characterized by elevated temperatures, humidity, and thermophilic microbes, limits its utility for household composting. This study aims to bridge the research gap in PLA’s recyclability and explore its feasibility in mechanical recycling processes. The research focuses on assessing the mechanical characteristics of PLA and PLA-based composites post-recycling. Specifically, we examined the effects of two extrusion methods—conical and parallel—on PLA and its composites containing 20 wt.% basalt fibers (BF). The recycling process encompassed repeated cycles of hot melt extrusion (HME), followed by mechanical grinding to produce granules. These granules were then subjected to injection moulding (IM) after 1, 3 and 5 recycling cycles. The tensile properties of the resulting IM-produced bars provided insights into the material’s durability and stability. The findings reveal that both PLA and PLA/BF composites retain their mechanical integrity through up to 5 cycles of mechanical recycling. This resilience underscores PLA’s potential for integration into existing recycling streams, addressing the dual challenges of environmental sustainability and waste management. The study contributes to the broader understanding of PLA’s lifecycle and opens new possibilities for its application in eco-friendly packaging, beyond the limits of composting. The implications of these findings extend towards enhancing the circularity of biopolymers and reducing the environmental footprint of plastic packaging. Full article
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16 pages, 3161 KiB  
Article
Screening, Characterization and Comparison of Endoglucanases/Xylanases from Thermophilic Fungi: A Thielavia terrestris Xylanase with High Activity-Stability Properties
by Shaohua Xu, Kexuan Ma, Zixiang Chen, Jian Zhao, Xin Song and Yuqi Qin
Int. J. Mol. Sci. 2025, 26(14), 6849; https://doi.org/10.3390/ijms26146849 - 17 Jul 2025
Viewed by 223
Abstract
Thermostable cellulases and xylanases have broad acceptance in food, feed, paper and pulp, and bioconversion of lignocellulosics. Thermophilic fungi serve as an excellent source of thermostable enzymes. This study characterized four endo-β-1,4-glucanases (two glycoside hydrolase (GH) family 5 and two GH7 members) and [...] Read more.
Thermostable cellulases and xylanases have broad acceptance in food, feed, paper and pulp, and bioconversion of lignocellulosics. Thermophilic fungi serve as an excellent source of thermostable enzymes. This study characterized four endo-β-1,4-glucanases (two glycoside hydrolase (GH) family 5 and two GH7 members) and four endo-β-1,4-xylanases (two GH10 and two GH11 members) from thermophilic fungus Thielavia terrestris, along with one GH10 endo-β-1,4-xylanase each from thermophilic fungus Chaetomium thermophilum and mesophilic fungus Chaetomium globosum. Comparative analysis was conducted against three previously reported GH10 endoxylanases: two thermostable enzymes from the thermophilic fungus Humicola insolens and thermophilic bacterium Halalkalibacterium halodurans, and one mesophilic enzyme from model fungus Neurospora crassa. The GH10 xylanase TtXyn10C (Thite_2118148; UniProt G2R8T7) from T. terrestris demonstrated high thermostability and activity, with an optimal temperature of 80–85 °C. It retained over 60% of its activity after 2 h at 70 °C, maintained approximately 30% activity after 15 min at 80 °C, and showed nearly complete stability following 1 min of exposure to 95 °C. TtXyn10C exhibited specific activity toward beechwood xylan (1130 ± 15 U/mg) that exceeded xylanases from H. insolens and H. halodurans while being comparable to N. crassa xylanase activity. Furthermore, TtXyn10C maintained stability across a pH range of 3–9 and resisted trypsin digestion, indicating its broad applicability. The study expands understanding of enzymes from thermophilic fungi. The discovery of the TtXyn10C offers a new model for investigating the high activity-stability trade-off and structure-activity relationships critical for industrial enzymes. Full article
(This article belongs to the Section Macromolecules)
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21 pages, 1897 KiB  
Article
Simulation of Conventional WWTPs Acting as Mediators in H2/CO2 Conversion into Methane
by Rubén González and Xiomar Gómez
Environments 2025, 12(7), 245; https://doi.org/10.3390/environments12070245 - 16 Jul 2025
Viewed by 509
Abstract
CO2-biomethanation was studied in the present manuscript by considering the direct injection of hydrogen into a conventional anaerobic digester treating sewage sludge within a simulated wastewater treatment plant (WWTP). The plant was simulated using the Python 3.12.4 software, and a Monte [...] Read more.
CO2-biomethanation was studied in the present manuscript by considering the direct injection of hydrogen into a conventional anaerobic digester treating sewage sludge within a simulated wastewater treatment plant (WWTP). The plant was simulated using the Python 3.12.4 software, and a Monte Carlo simulation was conducted to account for the high variability in the organic content of the wastewater and the methane potential of the sludge. Two modes of operation were studied. The first mode involves the use of an anaerobic digester to upgrade biogas, and the second mode considers using the digester as a CO2 utilization unit, transforming captured CO2. Upgrading biogas and utilizing the extra methane to generate electricity within the same plant leads to a negative economic balance (first scenario). A hydrogen injection of 1 L of H2/Lr d (volumetric H2 injection per liter of reactor per day) was required to transform the CO2 present in the biogas into methane. The benefits associated with this approach resulted in lower savings regarding heat recovery from the electrolyzer, increased electricity production, and an additional oxygen supply for the waste-activated sludge treatment system. Increasing the injection rate to values of 5 and 30 L of H2/Lr d was also studied by considering the operation of the digester under thermophilic conditions. The latter assumptions benefited from the better economy of scale associated with larger installations. They allowed for enough savings to be obtained in terms of the fuel demand for sludge drying, in addition to the previous categories analyzed in the biogas upgrading case. However, the current electricity price makes the proposal unfeasible unless a lower price is set for hydrogen generation. A standard electricity price of 7.6 c€/kWh was assumed for the analysis, but the specific operation of producing hydrogen required a price below 3.0 c€/kWh to achieve profitability. Full article
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15 pages, 2643 KiB  
Article
Hot Mineral Water as a Medium for Molecular Hydrogen Reactions in the Primordial Hydrosphere for the Origin of Life
by Ignat Ignatov, Teodora P. Popova, Paunka Vassileva, Yordan G. Marinov and Mario T. Iliev
Hydrogen 2025, 6(3), 48; https://doi.org/10.3390/hydrogen6030048 - 15 Jul 2025
Viewed by 1442
Abstract
Studies have been conducted on the potential development of Hydrogenobacter thermophilus and Pseudomonas aeruginosa in an anaerobic environment, both in the presence and absence of molecular hydrogen (H2). H. thermophilus developed better at 70 °C and pH 7.0 in the presence [...] Read more.
Studies have been conducted on the potential development of Hydrogenobacter thermophilus and Pseudomonas aeruginosa in an anaerobic environment, both in the presence and absence of molecular hydrogen (H2). H. thermophilus developed better at 70 °C and pH 7.0 in the presence of molecular hydrogen. It also multiplied in its absence, but to a lesser extent. Dissolved hydrogen in an amount of 1 ppm is biologically active for this thermophilic chemolithotrophic species. The tested strains of P. aeruginosa also showed growth under anaerobic conditions in the presence of H2 concentrations of 1 ppm and 2 ppm, which was ensured by adding Mg. The results indicate that not only the oldest microorganisms on our planet, archaebacteria, but also current species such as H. thermophilus and P. aeruginosa are capable of development under conditions characteristic of the ancient hydrosphere. DFT analyses showed that hydrogen water forms stable water clusters, whose hydrogen bond network retains and stabilizes reducing agents such as molecular hydrogen and magnesium (Mg0). This creates a microenvironment in which key redox processes associated with autotrophic growth and chemical evolution can occur. This is a realistic model of the Earth’s primordial hydrosphere’s conditions. Full article
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16 pages, 2368 KiB  
Article
Isolation and Characterization of a Thermaerobacillus caldiproteolyticus-like Strain Producing Extracellular Amylase from the Nelumwewa Geothermal Spring, Sri Lanka
by Sarath Bandara, Buddhika Dharmasena, Lakshani Pathirana, Prasad Jayasooriya and Aruna Weerasooriya
Fermentation 2025, 11(7), 397; https://doi.org/10.3390/fermentation11070397 - 11 Jul 2025
Viewed by 461
Abstract
The growing demand for efficient sustainable biocatalysts for industrial applications has driven the exploration of extremozymes from extremophiles, particularly those thriving in geothermal environments. This study aimed to isolate and characterize extracellular amylase-producing thermophilic bacteria from the Nelumwewa geothermal spring in Sri Lanka, [...] Read more.
The growing demand for efficient sustainable biocatalysts for industrial applications has driven the exploration of extremozymes from extremophiles, particularly those thriving in geothermal environments. This study aimed to isolate and characterize extracellular amylase-producing thermophilic bacteria from the Nelumwewa geothermal spring in Sri Lanka, an underexplored ecosystem. Among the isolated thermophilic bacterial strains, NW2 isolates exhibited a prominent extracellular amylase activity. Molecular characterization by 16S rRNA gene sequencing confirmed the close phylogenetic relationship between NW2 and Thermaerobacillus caldiproteolyticus, which is well-known for thermostable proteases. Biochemical assays revealed optimal amylase activity of NW2 isolate at 60 °C and pH 8.0, with a crude enzyme activity of 0.85 U/mL. The enzyme demonstrated efficient hydrolysis of raw cassava starch, highlighting its potential for industrial applications in food, biofuel, and detergent industries. This study reports the first T. caldiproteolyticus-like strain from Sri Lanka with significant extracellular amylase activity, emphasizing the biotechnological potential of geothermal springs as sources of novel extremozymes. These findings contribute to the growing repository of thermostable enzymes, highlighting the need for further exploration of Sri Lanka’s geothermal microbial diversity for industrial biocatalysts. Full article
(This article belongs to the Section Microbial Metabolism, Physiology & Genetics)
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16 pages, 1097 KiB  
Review
Beyond Fish Pathogens: A Comprehensive Overview of Aeromonas salmonicida
by Xiaotong Qin, Zhongduo Li, Jinglan Guo, Feng Bai and Xiaodong Ling
Microbiol. Res. 2025, 16(7), 157; https://doi.org/10.3390/microbiolres16070157 - 8 Jul 2025
Viewed by 395
Abstract
Aeromonas salmonicida is an age-old fish pathogen widely distributed in seawater and freshwater environments that causes significant economic losses to the global aquaculture industry. Genetic mutations and the emergence of thermophilic strains are factors in the continuous expansion of A. salmonicida’s host [...] Read more.
Aeromonas salmonicida is an age-old fish pathogen widely distributed in seawater and freshwater environments that causes significant economic losses to the global aquaculture industry. Genetic mutations and the emergence of thermophilic strains are factors in the continuous expansion of A. salmonicida’s host range. Beyond infecting fish, A. salmonicida poses a potential threat to mammalian and human health. This review synthesizes recent global research advances concerning A. salmonicida, encompassing strain characteristics, genomic features, virulence factors, and pathogenic mechanisms, as well as the clinical manifestations in infected fish and mammals, and discusses prevention and treatment methods. Particular emphasis is placed on evaluating the potential prophylactic roles of Chinese herbs and bacteriophages against A. salmonicida infection. Furthermore, the review provides perspectives on future research directions, diagnostics, and disease management, informed by contemporary domestic and international studies on this pathogen. Full article
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23 pages, 3357 KiB  
Article
Enhancing the Thermostability of a New Tannase Through Rational Design and Site-Directed Mutagenesis: A Quality Improvement Strategy for Green Tea Infusion
by Hai-Xiang Zhou, Shi-Ning Cao, Chu-Shu Zhang, Mian Wang, Yue-Yi Tang, Jing Chen, Li-Fei Zhu, Jie Sun, Qing-Biao Meng, Jing Chen and Jian-Cheng Zhang
Beverages 2025, 11(4), 99; https://doi.org/10.3390/beverages11040099 - 1 Jul 2025
Viewed by 583
Abstract
Tea has become one of the most popular drinks worldwide thanks to its pleasant sensory attributes and diverse health benefits. However, tannin-rich compositions have several negative effects and significantly impact the quality of tea beverages. Among various detannification methods, tannase treatment appears to [...] Read more.
Tea has become one of the most popular drinks worldwide thanks to its pleasant sensory attributes and diverse health benefits. However, tannin-rich compositions have several negative effects and significantly impact the quality of tea beverages. Among various detannification methods, tannase treatment appears to be the most secure and environmentally friendly strategy. Although numerous microbial tannases have been identified and used in food processing, they are predominantly mesophilic with compromised heat tolerance, which limit their application in high-temperature tea extraction processing. Computer-assisted rational design and site-directed mutagenesis has emerged as a promising strategy in enzyme engineering to improve the thermostability of industrial enzymes. Nevertheless, relevant studies for tannase thermostability improvement remain lacking. In the present study, a novel thermophilic tannase called TanPL1 from marine fungus Penicillium longicatenatum strain SM102 was expressed in the food-grade host Yarrowia lipolytica. After purification and characterization, the thermostability of this enzyme was improved through site-directed mutagenesis guided by computer-aided rational design and molecular dynamics simulations. Then the thermostable mutant MuTanPL1 was applied in green tea processing for both polyphenol extraction and ester catechin hydrolysis. The tannase yield and specific activity values of 166.4 U/mL and 1059.3 U/mg, respectively, were achieved. The optimum pH and temperature of recombinant TanPL1 were determined to be 5.5 and 55 °C, respectively, and the enzyme exhibited high activity toward various gallic acid ester substrates. The site-directed mutagenesis method successfully generated a single-point mutant, MuTanPL1, with significantly enhanced thermostability and a higher optimum temperature of 60 °C. After 2 h of detannification by MuTanPL1, nearly all gallated catechins in green tea infusion were biotransformed. This resulted in a 202.4% and 12.1-fold increase in non-ester catechins and gallic acid levels, respectively. Meanwhile, the quality of the tea infusion was also markedly improved. Sensory evaluation and antioxidant activity assays revealed notable enhancements in these properties, while turbidity was reduced considerably. Additionally, the α-amylase inhibition activity of the tannase-treated tea infusion declined from 50.49% to 8.56%, revealing a significantly lower anti-nutritional effect. These findings suggest that the thermostable tannase MuTanPL1 holds strong application prospects in tea beverage processing. Full article
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