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Investigating fungal gene expression in complex solid-state fermentation (SSF) systems is essential for process optimization but is hindered by challenges in RNA extraction. RNA extraction from fungal samples rich in plant biomass is a special challenge that is not met by the commercial RNA extraction kits and conventional RNA extraction methods we have tested. This study presents an optimized RNA extraction method, suitable for RT-qPCR from agricultural side streams fermented by filamentous fungi. The method was applied to investigate the temporal expression of the class II hydrophobin NC2 in Neurospora intermedia grown on four different sources of brewer's spent grain (BSG): IPA, Pilsner, Stout, and a microbrewery batch. Relative NC2 expression showed a clear temporal increase, peaking on day 6, with the microbrewery BSG batch having the highest peak expression. It demonstrates the applicability of the presented RNA extraction method to monitor expression of a gene of interest during solid-state fermentation of a complex substrate. Furthermore, the RNA extraction protocol was successfully validated on other challenging substrates, including rapeseed press cake and oat hulls, and with other fungi (Aspergillus oryzae and Trichoderma asperellum), consistently yielding high-purity RNA. This work provides a reliable method for RNA extraction from complex SSF products that can be used for gene expression analysis. KEY POINTS: • Reliable RNA extraction protocol developed for complex fungal SSF substrates • Hydrophobin NC2 expression in N. intermedia might be temporally regulated on BSG • Peak NC2 expression (day 6) was observed on BSG from a microbrewery.

The excessive reliance on synthetic fertilizers and pesticides has undeniably enhanced crop productivity; however, it has also led to long-term environmental degradation, highlighting the urgent need for sustainable alternatives. Bioinoculants, particularly siderophore-producing fungi, represent a potential eco-friendly approach to improving nutrient acquisition, plant health, and resistance to pathogens. In the present study, 23 fungal species were isolated from agricultural and non-agricultural soils, among which four exhibited plant growth-promoting fungal (PGPF) activity. Of these, Aspergillus awamori (SA2) and Aspergillus terreus (SA3), identified through ITS gene sequencing, demonstrated the highest PGPF potential. Both strains synthesized hydroxamate-type siderophores, as confirmed by Chrome Azurol S (CAS) assay, HPLC, FTIR, and NMR analyses. In addition, they exhibited multiple plant growth-promoting (PGP) traits, including indole-3-acetic acid and ammonia production, as well as phosphate solubilizing activity. Treatment of Vigna unguiculata (L.) Walp. seeds with the filtrate of siderophore- producing isolates significantly enhanced germination rate, biomass accumulation, chlorophyll content, and levels of secondary metabolites (phenolics and flavonoids), while also displaying antagonistic activity against Fusarium oxysporum and Rhizoctonia solani under controlled conditions. These findings suggest that Aspergillus awamori and Aspergillus terreus have potential as plant growth-promoting and biocontrol agents; however, further validation under field conditions is required.

Several homologous morphological characters, despite sharing apparently similar features, are known to have independently evolved in different lineages multiple times. However, the genetic backgrounds of such morphological convergences remain poorly understood. To detect any correlated amino acid substitutions potentially responsible for morphological convergence at the phenotypic level, we focused on the morphology of the septal pore cap (SPC), a structure involved in mycelia's complex multicellularity in fungi. SPCs are classified into three morphological types: perforate, imperforate, and vesiculate. To understand the evolutionary events that occurred at the sequence level during the morphological convergence of perforate SPCs in Agaricomycotina, we examined sequence differences among species with different SPC types by comparative genomic analysis using single-copy gene dataset from twelve Agaricomycotina genomes with morphological literature of SPC. Our analysis revealed that sequences of eight genes, including an SPC-related gene spc33, were clustered based on SPC morphology rather than species relationship. Additionally, same amino acid substitutions independently occurred in both lineages in which species with perforate SPCs emerged. These findings suggest that specific amino acid substitutions in spc33 were critical for the emergence of perforate SPCs in multiple lineages. Further, our gene search for spc33 across organisms suggests that spc33 evolved shortly before the emergence of imperforate SPC. This study represents the first step toward elucidating the genetic basis of the morphological evolution of SPC. It contributes to both clarifying the genetic basis underlying morphological convergence and advances the study of fungal evolutionary morphology.

Nannizziopsis arthrosporioides, a keratinophillic fungus, has caused infection in multiple reptile species. This study aimed to investigate the efficacy of commercially available disinfectants against the growth of N. arthrosporioides. Two molecularly confirmed isolates of N. arthrosporioides were used. Hyphal growth was collected from agar plates, filtered, and then diluted to high and low conidial concentrations. Each conidial suspension was exposed for 5 or 10 minutes to sterile water (control), 10% dilution of commercial bleach containing sodium hypochlorite, 409 multipurpose cleaner containing benzyl ammonium chloride, chlorhexidine 2% solution, F10 SC (benzyl ammonium chloride and polyhexanide) at recommended product labels (1:100, 1:250), 10% dilution of povidone-iodine, and Rescue disinfectant (activated hydrogen peroxide, 1:16). Both isolates at each exposure time failed to grow after exposure to 409 multipurpose cleaner, commercial bleach, chlorhexidine, povidone iodine, and Rescue disinfectant. Inhibition of N. arthrosporioides growth was variable following exposure to F10 SC. Nannizziopsis arthrosporioides can be effectively inactivated using various commercially available disinfectants, and data provided here adds to the growing body of knowledge surrounding appropriate disinfection of fungi that can act as reptile pathogens.

Porphyromonas gingivalis is an oral bacterium commonly associated with periodontitis. P. gingivalis synthesizes sphingolipids (SLs), and recently, we reported that P. gingivalis SLs packaged into outer membrane vesicles (OMVs) can regulate THP-1 macrophage inflammatory responses to P. gingivalis OMVs. The contribution that P. gingivalis SLs have on host-pathogen interactions remains poorly understood, especially in the context of OMVs. Here, we demonstrate that P. gingivalis SLs significantly reduce the uptake of OMVs isolated from SL-containing wild-type (WT) P. gingivalis compared to OMVs from an SL-null mutant P. gingivalis strain, and that the loss of SLs drives uptake of P. gingivalis OMVs via lipid rafts in THP-1 macrophages. Intriguingly, we found that the sensing of P. gingivalis OMVs via TLR2 and MyD88 is attenuated due to the presence of SLs. Lastly, transcriptomic analysis of THP-1 macrophages co-cultured with either WT or SL-null P. gingivalis OMVs for 2 h revealed an array of differentially expressed genes. Interestingly, at 2 h, WT P. gingivalis OMVs promoted an overall trend of gene downregulation in THP-1 macrophages when compared to unchallenged, whereas SL-null OMVs strongly promoted upregulated gene expression. These findings provide early-phase characterization of the role of SLs in initial host cellular responses to P. gingivalis OMVs and expand our knowledge of interactions between SL-containing P. gingivalis OMVs and the host.

Antibiotic resistance is one of the most important problems threatening global public health by complicating the treatment of infections worldwide. The increase in resistant microorganisms creates a serious economic and social burden on healthcare systems and increasingly limits treatment options. Conscious use of antibiotics, infection control measures and the development of alternative treatment strategies are vital to ensure sustainability in health. Klebsiella pneumoniae is the most common gram-negative bacterium among urinary tract infections. Treatment of infections has become difficult due to the resistance to beta-lactam antibiotics. Bee venom (BV) and nanovesicle fractions isolated from BV are bioactive compounds with antimicrobial and antibiofilm activity. The aim of this study was to determine the antimicrobial and antibiofilm effects of BVand bee venom-derived nanovesicle fractions against the nosocomial infection agent K. pneumoniae, and to evaluate CTX-M PCR band detection under the tested conditions. Minimum inhibitory concentration (MIC), antibiofilm activity, fractional inhibition concentrations (FIC), CTX-M PCR band detection, and viability rates in L929 cells were evaluated for both BVand the bee venom-derived nanovesicle fraction against K. pneumoniae. MIC value of nanovesicle fractions isolated from BVwas determined as 1.95 mg/L. In combination with piperacillin and tazobactam, a synergistic effect was detected with a value of 0.5. Antibiofilm activity was measured with the highest absorbance value of 0.163 and 0.094 for BV nanovesicle fractions. A detectable CTX-M PCR band was observed in the BVgroup at 0.5× MIC, whereas no detectable band was observed at both concentrations by day 4. In the bee venom-derived nanovesicle fraction group, no detectable CTX-M PCR band was observed at 2× MIC and 0.5× MIC under the tested conditions. In contrast, detectable CTX-M PCR bands were observed in the piperacillin-tazobactam group. It was reported that the combination groups decreased the viability rate in L929 cell lines. The bee venom-derived nanovesicle fraction showed antimicrobial, antibiofilm, and PCR-based detection findings under the tested conditions, supporting the need for further investigation in broader in vitro and in vivo models.

Growing interest in natural formulations for food and health applications has intensified research on probiotic lactic acid bacteria (LAB). In this study, a LAB strain designated MIC-3 was isolated from traditional curd and evaluated using phenotypic, safety, and whole-genome approaches. The isolate exhibited typical LAB characteristics, being Gram-positive and catalase-negative with broad carbohydrate fermentation ability. MIC-3 demonstrated moderate broad-spectrum antimicrobial activity, with inhibition zones up to 8.7 mm against selected pathogens. The strain was susceptible to clinically relevant antibiotics and displayed γ-hemolytic activity, confirming its safety profile. It maintained viability under acidic conditions and tolerated bile salt concentrations up to 0.8%. Adhesion-associated properties were significant, with 85% auto-aggregation, strong co-aggregation with pathogens, and high cell surface hydrophobicity. Whole-genome sequencing identified the strain as Lactiplantibacillus plantarum and revealed genes supporting probiotic functionality, including stress tolerance (atp operon, groEL/groES, dnaK, clp), bile resistance (bsh/cbh, ABC transporters), adhesion (srtA, slpA, eps clusters), and bacteriocin production (pln cluster). Genes involved in lactic acid production, carbohydrate uptake, and vitamin biosynthesis were also detected, while no virulence-associated genes were identified. Collectively, these findings confirm the probiotic and functional potential of L. plantarum MIC-3 for food and health applications.

The monophasic Salmonella Typhimurium variant (1,4,[5],12:i:-) is a growing global public health and food safety concern. This study analyzed 60 such isolates collected in Huzhou (2023-2025), including 16 outbreak strains. All isolates belonged to ST34 and carried 211 virulence genes. Antimicrobial susceptibility testing showed universal resistance to tetracycline (100%) and a high multidrug resistance rate (96.67% resistant to ≥ 3 antibiotic classes). Whole-genome sequencing identified 39 resistance genes; aac(6´)-Iaa was present in all strains, and 48.33% carried ≥ 4 resistance genes. Phylogenetic analysis revealed high overall homology with localized evolutionary traits. Notably, clinical outbreak and food-derived isolates were clonally identical (0 SNP difference).

Solar salt pans are extreme hypersaline environments that represent functionally specialised microbial communities mediating essential biogeochemical transformation. Vedaranyam, a coastal region of the Bay of Bengal containing artificially constructed solar salterns for salt production. There is limited information available on the metagenome diversity and functional profiling of this saltpan, which prompted us to investigate it. Here, we report the first whole metagenome sequencing to explore the dynamics of the functional structure of microbial communities in saltpan during the preharvest and postharvest phases of salt production. Methanobacteriota and Pseudomonadota dominated both phases at the phylum level, while Halobacteria comprised the most abundant class (53.2% preharvest; 48% postharvest). A notable bloom of Dactylococcopsis salina was observed during postharvest (4.28% to 12.67%) and flock doubling of Cyanobacterota relative abundance (5.5% to 10.6%), reflecting photosynthetic primary production following salt removal. Conversely, during postharvest phase sulfur oxidising Guyparkeria halophila reduced 23 fold, while the DMSP accumulating osmolyte producer Salinibaculum marinum dominated preharvest (6.98%). However, functional classification of the metagenome revealed active participation of the microbial community across five major biogeochemical cycles. Encompassing carbon fixation by cyanobacteria and diverse haloarchaea, nitrogen cycling through diazotrophy and denitrification, a cryptic preharvest sulfur cycle coupling sulfate reduction and sulphide oxidation, phase shifted DMSP catabolism, and light driven bacteriorhodopsin through archaeal energy conservation. Metagenomic assembly yielded ten metagenomic assembled genomes (MAGs), revealing the taxonomic diversity and metabolic potential of the dominant halophilic community across biogeochemical cycles. These results provide critical insights into the ecological succession from an anaerobic, chemolithotrophy-rich preharvest microbial community to an aerobic, photosynthetically driven postharvest assemblage, advancing our understanding of microbial biogeochemistry in managed hypersaline ecosystems.

Pseudolactococcus laudensis (formerly named Lactococcus laudensis) is an emerging lactic acid bacterium first isolated from raw milk in 2015 and subsequently detected in vegetables and dairy mesophilic starter cultures. Despite its recurrent isolation from diverse environments, the genetic basis of its niche adaptation, horizontal gene transfer and phage defence remains unexplored. Here, we perform the first comparative genomic and epigenomic analysis of P. laudensis using complete genomes of a plant-derived isolate (MCRI-603), a milk isolate (DSM 28961) and 20 strains from a Danish dairy mesophilic starter culture. Genomes were annotated and analysed using pangenomics, Clustering of Orthologous Genes and methylome profiling. Average nucleotide identity, pangenome and Clustering of Orthologous Genes analyses revealed niche-associated structure: dairy starter strains formed a tight cluster, while the plant isolate MCRI-603 and milk isolate DSM 28961 were more similar to each other than to the starter culture group. The pangenome comprised 4,946 genes, with 1,396 core genes. Dairy starter strains showed markedly elevated numbers of insertion sequences, pseudogenes, plasmids and genomic islands relative to MCRI-603, which was plasmid-free and carried very few insertion sequence elements or genomic islands. DSM 28961 displayed pseudogene count similar to the dairy starter strains but markedly fewer transposases. These patterns are consistent with a plant-associated origin of P. laudensis and progressive dairy specialization via mobile genetic element acquisition. The P. laudensis mobilome was found to carry key niche-related traits. Lactose utilization operons were plasmid-encoded, whereas exopolysaccharide-encoding loci, opp oligopeptide transport systems and several defence loci, including clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas), were consistently encoded within chromosomal integrative elements. All strains harboured prophage-like elements, including putatively intact prophages in 13 of them, and ~67% of 238 predicted antiphage systems resided on mobile genetic elements, underscoring their central role in phage defence. Restriction-modification systems dominated the defensome, and three strains encoded CRISPR-Cas systems (including type III-A and type I-C), indicating a higher prevalence than has been reported for Lactococcus lactis and Lactococcus cremoris, where CRISPR-Cas has rarely been observed. Methylome analysis identified 43 distinct motifs, of which 25 were novel. The P. laudensis methylome was overwhelmingly dominated by N⁶-methyladenine, and most motifs were short, non-palindromic and largely associated with type III restriction-modification systems and some type I and II subtypes. Nearly all strains exhibited distinct methylation profiles, including those isolated from the same dairy starter culture, highlighting extensive epigenetic diversification in dairy environments. Altogether, the data reveals a highly dynamic genomic and epigenomic landscape in P. laudensis, greatly shaped by mobile genetic elements, and provides a foundation for future work in this species and other Pseudolactococci.

Antimicrobial resistance represents a paramount challenge to global public health in the 21st century. The multidrug-resistant fungal pathogen Candida auris poses a critical and escalating threat to global public health. Characterized by rapid nosocomial transmission, persistent environmental contamination, and resistance to multiple antifungal classes, C. auris challenges healthcare systems worldwide. Its independent emergence across distinct geographic clades and exponential rise in cases, exacerbated by the COVID-19 pandemic, underscore the urgent need for robust, coordinated response. This review synthesizes the current knowledge on C. auris with a focus on its implications for public health policy, particularly in the European and Balkan healthcare settings, where surveillance gaps and cross-border transmission risks remain pronounced. We analyze the key drivers of spread, including diagnostic misidentification, extensive antifungal resistance, and lapses in infection control, and evaluate the strain on surveillance and hospital preparedness. Effective mitigation is fundamentally dependent on implementing comprehensive, multi-faceted infection prevention and control strategies, guided by antifungal stewardship and rapid diagnostics. We conclude that addressing the C. auris threat requires an urgent, coordinated international and regional response focused on strengthening surveillance networks, standardizing diagnostic and infection prevention and control protocols, and fostering data sharing across borders to contain this resilient pathogen.

Two Gram-stain-positive rod-shaped anaerobic bacterial strains were isolated from pig faeces and designated as strains YH-ros2226T and YH-ros2228. Phylogenetic analysis using 16S rRNA gene sequences revealed that the isolates were most closely related to Pararoseburia lenta KCTC 15957T, with 92.2% similarity. The multi-locus sequence tree revealed that the isolates formed a distinct cluster within the family Lachnospiraceae. The average nucleotide identity, digital DNA-DNA hybridization, average amino acid identity and percentage of conserved proteins values between the isolates and related strains within the family Lachnospiraceae ranged from 67.0% to 68.8%, 18.5% to 33.9%, 51.5% to 54.6% and 33.6% to 44.1%, respectively. The major cellular fatty acids were C16 : 0, C18 : 1  ω9c and C16 : 0 N alcohol. The cell wall peptidoglycan contained meso-diaminopimelic acid. The genomic DNA G+C contents of the strains were 46.8-47.0 mol%. The chemotaxonomic, phenotypic and phylogenetic properties of YH-ros2226T (=KCTC 25944T=JCM 37835T) and YH-ros2228 (=KCTC 25945=JCM 37836) suggested that they represented a novel genus and species within the family Lachnospiraceae, for which the name Porcibacter vitabionis gen. nov., sp. nov. is proposed.

Wetlands are among the world's most significant ecosystems. Studying how different grazing intensities affect soil bacterial diversity and community assembly in alpine wetlands is crucial for their sustainable management. Based on 16S sequencing technology, this study analyzed the differences in the composition, structure, and community assembly mechanisms of bacterial communities under different grazing impacts on the Qinghai-Tibet Plateau. The research results showed that under the influence of grazing, there was no significant difference in the α-diversity of the soil bacterial community, but a certain degree of reorganization occurred at the phylum level of bacteria. At the same grazing intensity, the relative abundances of different bacterial phyla in the wetland were different, and at different grazing intensities, the relative abundances of the same phylum might have significant differences. Co-occurrence network analysis indicated that wetlands with medium grazing intensity had a higher degree of modularity. The results of the null model and neutral model showed that, with the increase in grazing intensity, the assembly process of the soil bacterial community gradually shifted from being dominated by deterministic processes to being dominated by neutral processes, with a decrease in heterogeneous selection and an increase in dispersal limitation.IMPORTANCEThis work innovatively clarifies grazing's regulatory effects on alpine wetland soil bacterial community assembly, revealing the deterministic-to-neutral shift with increasing grazing intensity and higher modularity under moderate grazing. It deepens understanding of "disturbance-microbial assembly" relationships, bridging environmental science, disturbance ecology, and microbiology. Practically, it provides a scientific basis for optimizing grazing management to maintain microbial function, crucial for sustainable management of Qinghai-Tibet alpine wetlands and similar ecosystems.

Surgical site infection (SSI) is a potentially devastating complication following surgery and can result in a significant burden to patients and healthcare providers. Our study reports the risk and putative risk factors for SSI after open reduction and internal fixation (ORIF) of long bone fractures. We report findings from a large multicentre observational cohort undertaken in 49 National Health Service (NHS) hospitals in England between January 2010 to March 2020. Patients were prospectively followed up during their inpatient stay and post-discharge to identify SSI meeting standardised case definitions within a year of surgery. A total of 22,073 long bone ORIF procedures were included, with a median patient age of 59.1 years (IQR 40.1-75.0). Of these, 236 (1.07%) developed an SSI, with a median time onset of 17 days (IQR 11.0-30.2 days). Just under half (43.6%) were deep incisional and 11.0% organ space SSIs. Of monomicrobial infections, meticillin-sensitive Staphylococcus aureus (44.1%) was the most common causative pathogen. An ASA score of 4 was particularly associated with an increased risk of infection, compared against patients with an ASA score of 1 (risk difference: 2.30 95% CI 1.22-3.38, p&#x2009;<&#x2009;0.001). Patients with SSI were more likely to have surveillance discontinued due to death than patients without SSI (RD:1.77, 95% CI -&#xa0;0.54-4.09, p&#x2009;=&#x2009;0.060). Our results highlight the risk of infection following surgery on long bone fractures and identify patient and procedure factors associated with elevated risk. Clinicians should focus on mitigating these risk factors to minimize harm due to SSIs.

Salmonella Enteritidis is a major causative agent of gastroenteritis and foodborne illnesses, posing significant therapeutic challenges due to the rise of multidrug-resistant (MDR) strains. The increasing prevalence of resistant isolates highlights the need for alternative strategies to improve treatment outcomes. Lytic bacteriophage therapy has emerged as a promising complementary approach to antimicrobials. This study aimed to investigate the synergistic bacteriostatic and bactericidal effects of a newly formulated cocktail comprising three distinct lytic bacteriophages combined with selected antimicrobials against the standard strain of Salmonella Enteritidis (ATCC 13076). Three distinct bacteriophages were isolated from poultry farm wastewater, purified based on their different plaque morphologies, and characterized by transmission electron microscopy (TEM), revealing short non-contractile tailed caudoviruses. Phage stability was evaluated across a pH range of 3 to 11 and temperatures from 4 to 50 &#xb0;C. The minimum inhibitory concentration (MIC) was determined using the macrodilution (tube dilution) method, and viable bacterial counts (CFU/mL) were measured at 0, 6, 12, and 24 h to assess the bacteriostatic and bactericidal effects on the bacterial strain. The phage cocktail combined with ciprofloxacin and ceftriaxone showed significant synergistic activity, resulting in reductions of 5.8 and 4.9 log CFU/mL, respectively, corresponding to up to a 75% reduction in MIC. In contrast, combinations with ampicillin and erythromycin demonstrated the least efficacy. The phage cocktail alone achieved a 3.2 log reduction in CFU/mL. The phage cocktail demonstrated enhanced antibacterial activity when combined with ciprofloxacin or ceftriaxone, achieving greater reductions in Salmonella Enteritidis counts compared to individual treatments. Minimal synergy was observed with ampicillin and erythromycin. Overall, the evaluated phage-antimicrobial combinations exhibited superior antibacterial effects within the scope of this study.

The goal of this work was to examine the effect of different solvents (Water, EtOH 70%, and acetone) on the phenolic composition, antioxidant, and antibacterial capacity of Moroccan Mentha aquatica L. leaf extract. To this end, HPLC-ESI-FULL-MS was used to characterize the extracts, while the Folin-Ciocalteu and aluminum trichloride techniques were used to evaluate the total phenolic and flavonoid contents. To assess the antibacterial capacity, the microdilution technique was performed to calculate the minimal inhibition concentration (MIC), and minimal bactericidal concentration (MBC). Phytochemical profiling revealed that the extracts were rich in bioactive constituents, particularly ferulic acid derivative, caffeoyl-protocatechuic acid derivative, quercetin, and diosmetin 7-O-beta-D-glucuronide. The hydroethanolic extract contained the highest levels of total phenolic (62.2&#x2009;&#xb1;&#x2009;1.2&#xa0;mg GAE/g DW) and flavonoid (29.15&#x2009;&#xb1;&#x2009;0.09&#xa0;mg QE/g DW) contents, exceeding those of the acetonic extract (22.2&#x2009;&#xb1;&#x2009;0.6 and 10.17&#x2009;&#xb1;&#x2009;0.07&#xa0;mg GAE/g DW, respectively) and the water extract (22.4&#x2009;&#xb1;&#x2009;0.6 and 10.9&#x2009;&#xb1;&#x2009;0.6&#xa0;mg QE/g DW, respectively). This extract also showed the strongest antioxidant effect, recording an IC50 of 0.060&#x2009;&#xb1;&#x2009;0.001&#xa0;mg/mL in the DPPH assay, and an EC50 of 80&#xa0;&#xb5;g/mL in the RP test. In addition, it shows a great total antioxidant capacity, reaching 75.1&#x2009;&#xb1;&#x2009;2.0&#xa0;mg EAA/g DW when compared to water and acetonic extracts (28.5&#x2009;&#xb1;&#x2009;1.4 and 21.1&#x2009;&#xb1;&#x2009;0.1&#xa0;mg EAA/g DW, respectively). The antibacterial potential ranges from 0.78&#x2009;&#xb1;&#x2009;0.05&#xa0;mg/mL to 12.6&#xa0;mg/mL. In-silico prediction highlighted diosmetin 7-O-beta-D-glucuronide, quercetin, and equisetumpyrone as the key contributors to antioxidant capacity, while quercetin, 2,3,8-Tri-O-methylellagic acid, and diosmetin 7-O-beta-D-glucuronide were involved in antibacterial activity.

Alternaria alternata is a destructive postharvest pathogen that causes fruit decay and produces multiple mycotoxins, including alternariol (AOH), alternariol monomethyl ether (AME), and tenuazonic acid (TeA), posing a serious threat to crop quality and food safety. Biocontrol stands out as a promising strategy to mitigate both fungal infection and mycotoxin production. This study investigated that Bacillus sp. Z26 effectively inhibits the mycelial growth and mycotoxin accumulation of A. alternata. Comparative transcriptomic analysis revealed that strain Z26 did not directly repress canonical mycotoxin biosynthetic gene clusters; instead, it caused extensive transcriptional reprogramming in amino acid and lipid metabolism, with peroxisome-associated pathways being prominently affected. Among the downregulated genes, AaPex13 was identified as a key peroxisome biogenesis factor potentially involved in the biocontrol process. Targeted deletion of AaPex13 significantly impaired vegetative growth, conidiation, stress tolerance, and pathogenicity of A. alternata. Notably, loss of AaPex13 almost abolished TeA biosynthesis, while only partially reducing AOH and AME accumulation, indicating distinct metabolic dependencies among these toxins. In parallel, Bacillus sp. Z26 directly and selectively degraded AOH and AME, but not TeA. In grape berry assays, strain Z26 reduced disease severity, lowered mycotoxin contamination, and induced host defense responses, as evidenced by enhanced PPO and PAL activities and increased trans-resveratrol accumulation. Collectively, these findings suggest that Bacillus sp. Z26 suppresses A. alternata through a multifaceted mechanism involving disruption of AaPex13-mediated peroxisome biogenesis, selective detoxification of AOH and AME, and activation of host resistance. This study highlights peroxisome biogenesis as a previously underappreciated vulnerability in A. alternata and provides a potential target for precision biocontrol of Alternaria-associated postharvest diseases and mycotoxin contamination.

Asymptomatic Plasmodium vivax infections sustain transmission and derail elimination efforts, yet these hidden reservoirs evade current diagnostics. Here, we integrated computational immunology with empirical serology to discover high-performance B-cell epitopes. From a library of 142 P. vivax proteins, we prioritized six antigens (AMA1, CSP, DBP, MSP1, MSP9, and P12), synthesized 64 epitopes, and screened them against serum from P. vivax-infected cases from the China-Myanmar border (CMB) via ELISA, with multiplex immunoassay validation across Brazil, the Solomon Islands, and Papua New Guinea samples. Rigorous assessment of specificity, sensitivity, and asymptomatic discrimination, along with antigenic conservation analyses in CMB clinical isolates and global homologues, yielded ten immunodominant epitopes. Four exhibited outstanding performance, namely, MSP9_603-609 (90.4% sensitivity, AUC = 0.980), AMA1_457-466 (AUC = 0.910), MSP9_458-464(AUC = 0.802), and MSP9_425-433 (AUC = 0.898), all with absolute specificity against P. falciparum (P&#x2009;<&#x2009;0.0001). Crucially, the MSP9_384-389 epitope distinguished asymptomatic infections (AUC&#x2009;=&#x2009;0.756, P&#x2009;<&#x2009;0.0005). Despite variable detection rates across regions (2.2-22.2%), with the highest rates in Brazil, all the epitopes remained 99-100% conserved globally, reflecting strong functional immunological constraints. These epitopes enable unprecedented detection of both symptomatic and asymptomatic P. vivax infections - addressing a critical surveillance gap - while their scalability and species specificity make them ideal for elimination campaigns, particularly in low-resource settings. This work lays the foundation for next-generation serological tools to accelerate vivax malaria eradication.

Vaccines based on purified antigens, recombinant proteins, and nucleic acid platforms increasingly depend on adjuvants to induce robust, durable, and appropriately polarized immune responses in humans. While classical adjuvants such as aluminum salts and oil-in-water emulsions have enabled the success of many licensed vaccines, their largely empirical design limits adaptability to emerging pathogens and population-specific needs. This review presents a translational framework for next-generation vaccine adjuvant development by integrating nanotechnology-based delivery systems, innate immune signaling mechanisms, and systems-level computational strategies relevant to human vaccination. We summarize the mechanisms and clinical relevance of licensed and advanced adjuvants, including alum, MF59, AS01/AS04, saponins, toll-like receptor agonists, and lipid nanoparticles, with emphasis on influenza, HPV, herpes zoster, and COVID-19 vaccines. By linking immunological mechanisms with delivery engineering and predictive modeling, this review highlights rational strategies to support safer and more effective human vaccines.