Natural killer (NK) cells and natural killer T (NKT) cells contribute to the immune defense against hepatitis C virus (HCV). However, chronic hepatitis C (CHC) develops when there is a dysregulation of both innate and adaptive immune mechanisms. This case-control study was conducted to evaluate the immunological and clinical profiles of patients with chronic hepatitis C (CHC) in Sana'a, Yemen. The work focused on measuring T cell (CD3+), NK cell (CD56+), and NKT like cells (CD3 + CD56+) populations, along with examining viral load, hematological and biochemical indicators, and exploring how these factors relate to patients' demographic and clinical characteristics. A total of 100 subjects was established for this investigation, with an equal distribution of 50 CHC and 50 matched healthy controls (HC). The evaluation of peripheral blood samples encompassed multiple domains: serology (anti-HCV antibodies and HCV RNA viral load), biochemical liver function tests (measuring ALT, AST, ALP, and GGT levels), and a full hematological workup (CBC). Additionally, flow cytometric analysis was employed to determine the relative percentages of T, NK and NKT like cells in the CHC patient group and HC. The mean age of CHC patients was 48.0 ± 16.79 years, and most of them were > 50 years. There were 18 men (36.0%) and 32 women (64.0%). There was a significantly higher levels of ALT, AST, ALP, and GGT in CHC patients compared to those in HC (P < 0.05), This finding showed a significant decrease in the proportions of T and NKT like cells in CHC patients when compared to HC (P < 0.05). Furthermore, there was a significant negative correlation between the percentage of T and NKT like cells with total white blood cell (WBC) counts in patients with CHC (P < 0.05). In conclusion, our data demonstrated a reduction in the percentage of T, and NKT like cells among CHC patients in the study area. Thus, fewer immune cell responses may fail to clear HCV infections and may contribute to CHC development.

Poultry production remains significantly challenged by emerging and re-emerging avian viral diseases, which are influenced by host-pathogen interactions, viral evolution, and intensified farming systems. Viruses like avian influenza viruses (AIV) and Newcastle disease virus (NDV) remain the most persistent threats, while chicken anaemia virus (CAV), avian metapneumovirus (aMPV), adenoviruses, and astroviruses are increasingly recognized for their potential to affect poultry health and productivity. Despite the continued use of vaccination, biosecurity, and conventional diagnostics, effective control is often limited by delayed detection and inadequate integration of surveillance data. This review aims to critically examine the limitations of existing control strategies for major avian viral diseases and to evaluate the potential of artificial intelligence (AI) in improving disease surveillance and management. The current knowledge is synthesized on the basis of epidemiology and drivers of key viral infections and how AI-based tools can enhance predictive surveillance, real-time diagnostics, outbreak risk assessment, and vaccine development. The analysis indicates that AI-driven approaches can improve early detection and enable more effective use of heterogeneous data sources across animal, environmental, and public health domains. Integrating these approaches within a One Health framework offers opportunities to strengthen preparedness and response systems. Notwithstanding these benefits many issues regarding use of AI still need to be addressed for successful deployment of AI in diagnosing diseases.

This study investigated the therapeutic potential of the probiotic Roseburia intestinalis (R. intestinalis) in alleviating chronic kidney disease (CKD) symptoms and explored whether these effects are mediated through the modulation of the gut microbiota and fecal short-chain fatty acids (SCFAs). An Adriamycin (ADR)-induced CKD rat model was established, and the rats were treated with R. intestinalis via oral gavage for 8 weeks. Renal function, histopathology, and inflammatory markers were assessed. The gut microbiota composition was analyzed using 16 S rRNA sequencing, and SCFA levels were quantified by gas chromatography. To verify the underlying mechanism, fecal microbiota transplantation (FMT) was performed in germ-free CKD rats. The results demonstrate that R. intestinalis significantly mitigated CKD symptoms, including reduced proteinuria, serum creatinine, and renal inflammation, while reversing weight loss. The treatment restored gut microbial diversity, specifically by increasing the abundance of Firmicutes and fecal SCFA levels (acetate, propionate, and butyrate). Importantly, FMT from R. intestinalis-treated rats into germ-free CKD rats reversed these beneficial effects, whereas R. intestinalis alone under germ-free conditions was less effective, suggesting the need for a complex microbiome for full efficacy. These findings indicate that R. intestinalis is associated with alleviation of CKD symptoms by modulating the gut microbiota and enhancing fecal SCFA production, highlighting its potential as a promising probiotic intervention for CKD management and underscoring the role of the gut-kidney axis as a therapeutic target.

With the aim to harness synergistic potential of multiple beneficial microbes, we developed consortia (BC6) bioformulations that comprise six bacterial strains in multiple delivery forms (both powder and liquid). The bioefficacy of the consortia was assessed in terms of plant growth, fruit quality, productivity, stress response and metabolite profile in tomato fruits. The novel liquid bioformulation demonstrated superior shelf-life stability beyond 12 months. Bacillus sp. IIVRBT-12, a prominent member of consortia exhibited plant growth-promoting (PGP) traits including phosphate (13.84 mg P-released/100 mg tricalcium phosphate) and zinc (47.24 mg ZnCO3 L[- 1]) solubilization. Bacillus sp. IIVRBB-3 showed high indole acetic acid (IAA) production (59.14 µg ml[- 1]) and ACC deaminase (872.42 nmol α-ketobutyrate mg[-1]h[- 1]) activity. Three-stage concomitant application (seed priming, root dip, and soil treatment) significantly improved plant growth with BC6-L treated plants showing enhanced height (89.95 cm) and root development (36.95 cm) compared to BC6-P, although both performed significantly against the control plants. The treatments improved flowering time and increased flowers per cluster (5.25 in BC6-L compared to 3.5 in BC6-P treated plants). Treatments enhanced fruit yield from 3.1 kg in BC6-P to 3.72 in BC6-L per plant. The consortia also significantly improved activity of defense enzymes and enhanced biochemical parameters (carotenoids, flavonoids, and ascorbic acid) of fruit nutritional quality. LC-MS/MS based putative metabolomic characterization of metabolites uniquely identified scopoletin, deoxyribose-5-phosphate, 2'-deoxycytidine-5'-diphosphate (sodium salt), adenosine monophosphate, 2'-deoxyuridine-5'-monophosphate (disodium salt) and cytidine-5'-monophosphate monohydrate in BC6 liquid inoculated plants. These findings demonstrated potential of BC6 consortia as an effective plant growth promoter offering enhanced crop productivity, improved nutritional quality, and increased stress resilience.

Laccase, with its wide substrate range, has significant potential in industrial biocatalysis, where agricultural waste has attracted considerable interest as an economical and sustainable alternative as a low-cost source for enzyme production. In this study, the effect of various factors (different aromatic compounds, carbon and nitrogen nutrient sources, metal ions, pH range, incubation temperature and time, and agricultural wastes as potential low-cost substrates) on the thermophilic laccase production performance from Bacillus licheniformis SO7 strain (GenBank No: MG076977) were evaluated in detail. Accordingly, optimal conditions were determined using statistical methods such as one-factor-at-time (OFAT) and Plackett-Burman desings to achieve the highest laccase production yield. Under these conditions, the impact of aromatic substrates on laccase production was investigated at the gene and the protein levels using qPCR and SDS-PAGE, respectively. Maximum laccase activity was obtained after 36 h of incubation under the following conditions: pH 8.0, 55 °C, 50 g/L maltose, 1 g/L fish peptone, 2.5 mM syringic acid, and 1 mM ZnCl2. The highest gene expression and protein level were found in the syringic acid-containing medium, correlating with the activity results. Furthermore, even in bacterial culture media containing only carrot peels, similarly high laccase enzyme activity was detected as in other substrates. This clearly demonstrates that agricultural wastes offer an economical, environmentally sustainable, and efficient alternative for bacterial growth and target enzyme production.

This study aimed to evaluate the use of apple juice (AJ), an agro-industrial by-product rich in sugars, as a low-cost substrate for the production of Vishniacozyma victoriae, a biocontrol yeast, and to assess the performance of the fresh and lyophilized biomass under semi-commercial postharvest conditions. A Central Composite Design was used to optimise culture medium composition for improve biomass production. Biomass production was then evaluated under different cultivation modes, batch and semicontinuous reactors at laboratory scale. The biomass was preserved by lyophilization using protective formulations and later applied in two commercial packinghouses. In organic Packinghouse A, the control percentages exceeding 50% and, in some cases, reaching 100% control for the decays caused by Penicillium expansum, Botrytis cinerea and Alternaria-Cladosporium spp. The antagonistic yeast was able to colonize the surface of the fruit during the postharvest period. Apple juice is a suitable substrate for the cost-effective production of biocontrol yeast. Semicontinuous cultivation enhanced biomass yield, and both fresh and lyophilized formulations were effective in reducing postharvest decay in pears and apples under semi-commercial conditions.

Arbuscular mycorrhizal fungi (AMF) are known enhancers of plant abiotic stress tolerance, yet their responses to varying temperatures and subsequent effects on host physiology remain less understood. Here, long bean (Vigna unguiculata ssp. sesquipedalis) plants were cultivated under three temperature conditions 25 °C, 35 °C, and outdoors (fluctuating 24-43.5 °C) to assess AMF community diversity, plant root colonisation, phenotypic traits, physiological traits, and heat shock protein (hsp) gene expression. AMF-targeted amplicon sequencing and morphological analyses showed AMF diversity to be higher with elevated and fluctuating temperatures; outdoor conditions supported greater taxonomic diversity, including Acaulospora, Claroideoglomus, Diversispora, and Archaeospora, while Glomus remained dominant across all treatments. Among the growing conditions, root colonisation was highest at 25 °C. AMF inoculation significantly enhanced leaf number and chlorophyll content, and promoted antioxidant enzyme activities, with the strongest responses observed at 35 °C. In contrast, qRT-PCR analysis of 13 hsp revealed that AMF colonisation had little effect on expression with only Vuhsp26.5 showing significantly increased expression at 35 °C. Temperature influenced transcriptional patterns of three of the 13 hsp, with Vuhsp, Vuhsp22.7-1 and Vuhsp26.5 markedly upregulated at 35 °C compared to 25 °C and outdoor conditions. Collectively, these results demonstrate that AMF inoculation contributes to improved performance in V. unguiculata through enhanced antioxidant defence, while temperature exerts a dominant role in shaping both AMF diversity and hsp gene regulation. This study highlights the context-dependent nature of AMF plant interactions and provides insights into their potential role in improving crop resilience under warming climates.

In Asian countries, the decoction of Rhus chinensis berries is used in alleviation of gut related ailments, especially gastroenteritis. In the background of its ethnobotanical use, the aim of the study was to determine the inhibitory activity of berries against a gut pathobiont causing gut dysbiosis, namely Clostridioides difficile. The hot water extract of Rhus chinensis (RcHWE) indicated appreciable antimicrobial activity (60% to near 100%) against C. difficile ATCC 43,593, in the tested range of 0.5-5.5 mg/mL, evaluated by microbroth dilution and standard plate count technique. The minimum inhibitory concentration (MIC) was determined to be 3 mg/mL. There was visible inhibition of biofilm formation on filter membrane impregnated with varying concentration of RcHWE (0.25MIC - 10MIC). Results of flow cytometry with propidium iodide (PI) stained red fluorescence shift in extract supplemented cells, crystal violet staining with decrease in absorbance (34% to 100%) relative to unsupplemented control as well as confocal microscopy images (increase in PI-stained cells), across the tested range (0.25MIC- 10MIC), clearly indicated the antimicrobial and antibiofilm forming ability of RcHWE against C. difficile. Additionally, SEM results along with lactate dehydrogenase assay, clearly indicated extensive cell damage and destruction of cell integrity. Inhibition of p-cresol production (90-100%) with supplementation of RcHWE, was found to be both concentration and time dependent. UHPLC-HRMS/MS analysis indicated polyphenolic glycosides to be the predominant constituents of the extract. Findings highlights the modulatory potential of RcHWE against C. difficile and provides preliminary evidence for its use in gastric ailments especially due to gut dysbiosis.

The genome of Mycobacterium smegmatis encodes numerous penicillin-interacting enzymes, and analysing their functions provides insights into the evolutionary mechanisms behind beta-lactam resistance in mycobacteria. In this study, we characterised one such enzyme, MSMEG_6194, annotated as a putative beta-lactamase. Although MSMEG_6194 shares structural similarity with class A beta-lactamases, it showed no detectable beta-lactamase activity under the tested conditions. Heterologous expression of MSMEG_6194 in Escherichia coli and Δmsmeg_6194 deleted strains of M. smegmatis did not confer significant resistance to beta-lactams, and the purified protein failed to hydrolyse nitrocefin either. However, ectopic expression of MSMEG_6194 partially restores morphological defects in seven PBP-deleted E. coli strains, and the purified enzyme cleaves the terminal D-alanine from a pentapeptide substrate, supporting its DD-carboxypeptidase activity. Structural analysis revealed the absence of a conserved glutamic acid residue in the omega-loop, which is critical for beta-lactamase catalysis in class A beta-lactamase. Substituting this residue (V139E mutant) imparts beta-lactamase activity, though it significantly reduces DD-carboxypeptidase function. Overall, these findings suggest that MSMEG_6194 act as a DD-carboxypeptidase and illustrate how a single amino acid change can alter catalytic preference, shedding light on the evolutionary transition from DD-carboxypeptidases to beta-lactamases in mycobacteria.

Leishmania donovani, the causative agent of visceral leishmaniasis, possesses a highly plastic genome and relies extensively on post-transcriptional regulation, posing challenges for accurate genome annotation. Despite the availability of a high-quality reference genome, many protein-coding genes remain incomplete or misannotated. While proteogenomic workflows have been previously applied to other Leishmania species, their application to the L. donovani CL-SL strain, which exhibits distinct genomic and disease-associated features, remains limited. In this study, we utilised a proteogenomic strategy integrating high-resolution tandem mass spectrometry (MS/MS) data with a custom six-frame translated genome database to re-analyse and refine the strain-specific genome annotation of L. donovani. This study is based on the systematic re-analysis of previously published proteomic datasets originally derived from the L. donovani DD8 strain and cross-mapped to the closely related L. donovani CL-SL reference assembly. By utilising stringent selection criteria, this study enables the extraction of additional, high-confidence annotation-level insights beyond the scope of the original analyses, while remaining cautious of localised, strain-specific sequence variations. Re-analysing published proteomics identified 50 N-terminal extensions and 15 novel coding regions. Many show orthology in related species and contain functional domains linked to metabolism, trafficking, or stress responses. These findings represent structural refinements to existing gene models, awaiting further experimental validation. Collectively, our findings demonstrate that proteogenomic integration significantly improves the accuracy of L. donovani genome annotation by resolving truncated gene models and uncovering hidden coding potential. These findings improve genome annotation and highlight previously unrecognised coding potential supported by peptide evidence in Leishmania donovani.

Myxococcus xanthus is a Gram-negative bacterium with a complex life cycle that includes vegetative swarming and fruiting-body formation. Previous studies have shown that the exopolysaccharides (EPS) produced by M. xanthus was essential for the developmental process, but the information concerning EPS structure in M. xanthus was lacking. In this study, a novel soluble polysaccharide EPS15M was purified from the extracellular supernatant of M. xanthus DK1622, which was mainly composed of mannose, glucose, glucosamine, and rhamnose. Methylation analysis suggested the prevalence of terminal and 2-linked glycosyl residues, consistent with a branched structure. NMR and liquid chromatography-mass spectrometry analyses confirmed that EPS15M is an α-D-(1,2)-linked mannan backbone with α-D-(1,6)-linked mannose/glucose branches, a structure that differs from any extracellular polysaccharide reported to date from myxobacteria. Besides, EPS15M modestly improves the developmental sporulation of the csgA mutant (approximately 17% of the wild‑type level) at a concentration of 0.1 mg/mL. Collectively, our findings present the first high-resolution structure of a soluble exopolysaccharide from myxobacteria and demonstrate its partial functional compensation of csgA mutant, suggesting that EPS15M may be involved in C‑signal‑mediated developmental sporulation. These results open new avenues for understanding the regulatory role of polysaccharides in the myxobacterial life cycle.

This study examines a nanosuspension of secondary metabolites (NSM) derived from Streptomyces chrestomyceticus strain ADP4, previously reported for its antifungal efficacy against Candida spp., and extends its evaluation to WHO-listed high-priority bacterial pathogens. The antibacterial potential of the NSM was assessed against Staphylococcus aureus ATCC 25,923, Enterococcus faecium ATCC 49,924, and Enterococcus faecalis ATCC 29,212. Transmission electron microscopy revealed the formation of uniformly dispersed, spherical nanoparticles with an average diameter of 14 ± 4.26 nm. Compared with the crude secondary metabolites (SMs), the NSM demonstrated noticeably enhanced antibacterial performance, reflected by increased zones of inhibition (10%, 16%, and 26%), significant reductions in minimum inhibitory concentrations (53%, 42%, and 71%), and improved minimum biofilm inhibitory concentrations (41%, 30%, and 70%) against S. aureus, E. faecium, and E. faecalis, respectively. The observed enhancement in antimicrobial and antibiofilm activity highlights the role of nano-formulation in improving the bioavailability and biological efficiency of microbial SMs. Also, the flow cytometric analysis of bacterial cells treated with SMs and NSM revealed an increase in membrane permeability in a dose- and time-dependent manner. These findings highlight the potential of actinobacteria-derived NSM as environmentally relevant and support its applicability in macromolecular and surface-associated biomedical applications, including antibiofilm coatings for medical devices, wound dressings, and infection-resistant biomaterials making NSM as a potential strategy for sustainable and microbe-driven innovation.

Soil salinization seriously threatens global food security with continuously expanding affected areas. Bacillus megaterium-based bioinoculant serves as an eco-friendly strategy for saline-alkali land remediation, whereas wild-type (WT) strains generally exhibit poor salt-alkali tolerance. To address this limitation, atmospheric and room temperature plasma (ARTP) mutagenesis combined with a microbial microdroplet culture (MMC) high-throughput screening system was adopted to screen high salt-tolerant B. megaterium mutant followed by multi-omics characterization. Using WT B. megaterium LD (CGMCC No.21828) as the original strain, a mutant library was constructed via optimized ARTP treatment, and the elite mutant SLD48 was isolated under dynamically increasing salt gradients. Compared with the wild type, SLD48 possesses a larger genome of 6,252,538 bp and acquires 28 extra functional genes including spoIVCA and pJ_gene0003. Intracellular osmoprotectants (proline, glycine betaine) and antioxidant α-tocopherol were markedly accumulated in SLD48. moreover, exogenous supplementation of these substances in the culture medium can effectively alleviate the damage caused by salt stress to the cells. In pot trials with saline-alkali soil, the rhizosphere colonization of SLD48 reached 3.52 × 10[5] CFU/g (2.46-fold of LD), and maize shoot fresh weight increased by 40.5% after inoculation. Collectively, this work confirms that mutant SLD48 is a promising microbial resource for saline-alkali soil improvement and crop yield promotion.

Wound healing remains a significant clinical challenge due to susceptibility to bacterial infections, excessive oxidative stress and delayed cellular regeneration. The extensive and often indiscriminate use of antibiotics has further intensified the emergence of antimicrobial resistance. In this study, a multifunctional smart wound dressing is developed by integrating a chromogenic hydrogel with electrospun poly(ε-caprolactone) (PCL) fibers functionalized with quaternary ammonium moieties anchoring ionic silver. The PCL fibers functionalized with ionic silver provide antibiotic-free antibacterial properties, while the chromogenic hydrogel allows for real-time colorimetric detection of metabolically active microorganisms. The colorimetric assessment of the dressing in both in vitro and ex vivo environments has shown a distinct, time-dependent reaction to metabolically active bacterial populations. It has demonstrated outstanding cellular biocompatibility, increased cell viability in Vero cells, a notable decrease in oxidative stress, and enhanced cell migration in an in vitro scratch assay. This multifunctional dressing showcases a promising platform for advanced wound management applications by integrating antibacterial effects, real-time infection detection, and pro-regenerative cellular responses.

Resveratrol and its glycoside polydatin are high-value stilbene metabolites with broad pharmacological potential, but their natural accumulation in plants is usually limited. Endophytic fungal elicitation may provide a useful approach for improving the accumulation of these compounds. In this study, three crude elicitors prepared from the endophytic fungus Plectosphaerella sp. J-G was evaluated for its effects on seedling growth and stilbene accumulation in Rumex gmelinii Turcz. ex Ledeb. sterile seedlings. Among the tested elicitor types, concentrations, and treatment durations, the mycelial homogenate elicitor at 250 mg·L[-1] for 10 days showed a relatively strong induction effect. Under this condition, the total content of resveratrol and polydatin in roots increased by 272.50% compared with the time-matched 10-day sterile water control, from 750.60 to 2795.97 µg·g[-1]. This treatment was also accompanied by 22.02% higher plant height and 165.63% greater root fresh weight. Transcriptome sequencing and real-time quantitative PCR validation were used to examine transcriptional changes associated with this response. Eight upregulated genes, including IAA, CYP73A/C4H, LDH, SAUR, STS, AOC3, ADH1, and POD, were potentially associated with precursor supply, stilbene/phenylpropanoid biosynthesis, defense response, and auxin-related growth regulation. Several transcription factor families, such as MYB, AP2/ERF, and WRKY, may also be related to the transcriptional response under J-G crude elicitor treatment. These results suggest that J-G crude elicitor treatment may be a useful induction strategy for increasing resveratrol and polydatin accumulation in RGT sterile seedlings and provide candidate genes for future functional validation.