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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.

Subclinical mastitis (SCM) is characterized as inflammation of mammary gland with absence of clinical signs. S. aureus, one of the principal causes of disease is known for production of various virulence factors, including enterotoxins, biofilm production and antibiotic resistant genes. Therefore, the present study was carried out with the objective of molecular characterization for various virulence associated genes, antibiogram profile, and MLST typing of S. aureus from milk samples from various districts of Haryana state of India. In our study, milk samples from 1154 quarters affected with subclinical mastitis (by CMT) subjected to isolation of Staphylococcus spp. yielded 519 isolates. Of them, 352 isolates were identified as S. aureus by targeting 23 S rRNA gene by PCR. Various genes possessed by these isolates were: (13.35%) mecA (methicillin resistant gene), (11.64%) biofilm formation gene icaD, (11.36%) biofilm formation gene icaA, and (9.37%) toxic shocks syndrome gene (tsst). Enterotoxin genes detected were: sec in (8.52%), sea in (8.23%), sed in (4.54%), seb in (2.84%) and see in (1.7%). Exfoliative genes etb and eta were detected in (3.97%) and (3.12%) isolates, respectively. Antimicrobial sensitivity assay revealed sensitivity in descending order to gentamicin, doxycycline, sulfisoxazole, neomycin, ceftriaxone, cefoperazone, chloramphenicol, ciprofloxacin, enrofloxacin, vancomycin, clindamycin, linezolid, cefoxitin and least susceptible to oxytetracycline. The finding of 'Multilocus Sequence Typing' (MLST) showed high diversity of sequence types and clonal complex of S. aureus isolates.

The slender-horned gazelle (SHG, Gazella leptoceros) is a small, sand-colored antelope native to northern Africa. It is endangered due to poaching and habitat loss. Limited reports describe health or disease in wild or zoo populations. Mucosal plaques in the oral cavity (n = 2), papillomas in the rumen (n = 3) or reticulum (n = 1), and squamous cell carcinoma in the oral cavity (n = 3), rumen (n = 4), or reticulum (n=1) were seen independently or concurrently in eight of 74 SHG postmortem examinations in a zoological collection between 1983 and 2023. All tumors were diagnosed in adults. Polymerase chain reaction (PCR) testing for the papillomavirus (PV) L1 gene was positive in tumors from six of the eight SHG and negative in the same tissue types in SHG without tumors. Additional PCR testing and sequencing produced a 1041 bp contiguous consensus sequence of the papillomavirus L1 gene, and two complete PV genomes of 7354 and 7303 bp (51 bp deletion in the latter of one SHG tumor). BLASTN analysis of the L1 sequence (with highest query coverage, 96%) showed giant panda Ailuropoda melanoleuca PV1 as the closest match in GenBank (69.4% identity) to the SHG papillomavirus, which has been provisionally named Gazella leptoceros papillomavirus 1 (GlPV1); the variant with the deletion has provisionally been named GlPV1-variant (GlPV1-var). Bayesian (nucleotide) and neighbor joining (amino acid) phylogenetic analysis placed GlPV1 in a distinct clade with Ailuropoda melanoleuca PV1, Rickett's big-footed bat Myotis ricketti PV, and Polar bear Ursus maritimus PV. To the authors' knowledge, these are the first non-bovid papillomaviruses to be described in SHG. Our results contribute to a growing list of PVs that may play a role in the development of benign or malignant oral and gastric tumors, and disease processes to which SHGs are susceptible.

The archaeal and bacterial community was determined in a continuous aerobic nitrifying reactor maintained under similar conditions for > 9 years, applied solely with ammonium as energy source and carbonate as C source. The high-throughput shotgun analysis revealed 4483 bacterial and 245 archaeal species that thrived on the metabolites provided by the autotrophic nitrifying population, mostly the ammonia oxidizing bacteria Nitrosomonas europaea and archaea Candidatus Nitrosocosmicus exaquare, and the nitrite oxidizing bacteria Nitrobacter winogradskyi and N. hamburgensis. Candidatus Nitrospira inopinata capable of oxidizing ammonia to nitrate was also detected. Although the reactor was supplied with sufficient O2, the anaerobic bacteria Candidatus Kuenenia stuttgardiensis capable of oxidizing ammonium to dinitrogen gas using nitrite as the electron acceptor under anoxic conditions (anammox) and four archaeal genera of the strict Methanobacteriaceae were detected in the reactor. A wide range of genes encoding for the different processes involved in N cycling were detected including the hzo gene encoding for the hydrazine oxidoreductase participating in the anaerobic anammox process. It was found that a bioreactor applied only with ammonium as energy source and maintained for > 9 years under steady state conditions contained a highly diverse bacterial and archaeal population and a wide range of metabolic processes related to the N cycle, which has not been reported before. This provides us with comprehensive insights into the dynamics of microbial communities in these types of systems.

A detailed characterization of the microbial ecosystem involved in the production processes of fermented foods is essential. Although fermented foods are an important part of the human diet and have seen an increasing interest nowadays, some challenges still need to be solved. Specifically, yeast identification through culture-independent methodologies is still limited to the genus level. Unlike bacterial species identifications, long-read sequencing technologies have barely been used for yeast species identification, and, to the best of the authors' knowledge, it has not been validated with mock communities reflecting food fermentation processes yet. Therefore, in the current study, we present an amplicon-based metabarcoding approach targeting the full-length internal transcribed spacer (ITS) region comprising ITS1, the 5.8S rRNA gene, and ITS2, using the PacBio HiFi sequencing platform. This method was validated using DNA-based mock communities composed of yeast species involved in sourdough, lambic beer, and cocoa fermentation processes. Accurate species-level identification was achieved for most of the species. However, special attention should be given to Saccharomyces-rich niches, as accurate species-level identification for this genus is still challenging. Furthermore, underestimation of the relative abundance of species with short ITS regions, such as Pichia and Brettanomyces, occurred. In addition, the method was successfully applied to describe the yeast diversity present in two sourdough and two lambic beer samples. Overall, the current method provides an unprecedented way of determining the species-level yeast composition of complex ecosystems present in fermented food products.IMPORTANCETo date, species-level identification of common yeasts present in food fermentation ecosystems has been difficult, if not impossible, when using short-read sequencing methods. However, species-level identification is essential when evaluating and describing the characteristics of fermented food microbiomes. The current study reports on the development and validation of an amplicon-based metabarcoding approach combined with long-read PacBio HiFi sequencing targeting the full internal transcribed spacer (ITS) region, comprising the ITS1 and ITS2 regions, as well as the 5.8S rRNA gene. The described methodology enables species-level identification of the most common yeasts present in food fermentation ecosystems. This new methodology provides an important tool not only for the investigation of fermented foods but also for other fields engaged in complex microbial community analysis.

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.

Fe(II)/α-ketoglutarate-dependent dioxygenases (αKGDs) are versatile biocatalysts whose catalytic activity relies on an efficient supply of O2 and αKG. However, in engineering applications, the O2 supply is seldom considered, whereas αKG is typically supplied through the oxidation of l-glutamate by l-glutamate oxidase, which concomitantly produces H2O2 that potently inhibits αKGD activity. This study developed a tunnel engineering strategy based on random accelerated molecular dynamics and protein structuromics to precisely modulate the access of small molecules (O2 and H2O2) to isoleucine dioxygenase (IDO), enhancing O2 transport while resisting H2O2 transport. Notably, mutant 1 (L179C/V225F/I240V) and mutant 2 (N193S/V225I/I240V) displayed markedly higher catalytic activity for five aliphatic amino acid substrates (Leu, Nle, Nva, Met, and Ile), reaching 27.2-fold greater activity. In addition, mutant 2 exhibited 7.1-fold higher activity against Ile in the presence of H2O2. This study highlights O2 and H2O2 tunnel engineering as a new strategy for enhancing αKGDs activity and antioxidative properties.

Carbapenem-resistant Acinetobacter baumannii (CRAB) represents a critical global health threat for which existing antibiotics are increasingly inadequate. This study aimed to establish a comprehensive genomic framework for the rational prioritization of virulent Acinetobacter bacteriophages as therapeutic candidates. We performed large-scale comparative genomic analysis of 340 virulent Acinetobacter bacteriophages, integrating phylogenetic reconstruction, pangenome analysis, CRISPR spacer-based host interaction mapping, Anti-CRISPR protein identification, and systematic antimicrobial resistance (AMR) gene screening. Genome sizes spanned a nearly 20-fold range, with a significant negative correlation between genome size and GC content (R² = 0.139, ρ = -0.630). Phylogenetic analysis revealed extensive divergence across multiple lineages with no dominant clade. Pangenome analysis identified 20,982 unique protein families, of which 76.2% were cloud genes, confirming a highly open genome architecture. CRISPR spacer matching yielded 1,480 high-confidence matches across 100 phage genomes, providing molecular evidence of broad historical infectivity. Anti-CRISPR profiling identified Acinetobacter phage XC1 as an exceptional therapeutic candidate harboring 55 predicted Anti-CRISPR proteins with canonical regulatory locus architecture. AMR screening identified 21 distinct AMR gene homologs (Loose RGI hits, 22.5 to 47.1% amino acid identity) distributed heterogeneously across the dataset, confirming abundant therapeutically clean candidates while flagging a subset warranting further scrutiny before therapeutic exclusion. These findings provide a multi-criteria genomic framework for rational phage candidate prioritization against multidrug-resistant Acinetobacter infections, with direct implications for evidence-based phage therapy development.

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 °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.

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.

Osteoarthritis (OA) primarly involves the degradation of joint cartilage and requires new treatments. REJENERA©, a newly developed nutraceutical formula against OA, contains primarily olive leaf bioflavonoids ( ZeyEX©, quercetin and luteolin), S-allylcysteine, palmitoylethanolamide, L-proline, hyaluronic acid and boron. This study focuses on the efficacy of REJENERA in treating knee OA and aims to compare it with ZeyEX, NPROC© (a product obtained by combining collagen-rich eggshell membrane with olive leaf extract), and IBUPROFEN in a rat OA model. OA was established by intra-articular injection of monosodium iodoacetate (MIA; 3  mg) into the right knee joints. Rats were either left untreated or treated orally for 12 weeks with REJENERA, ZeyEX, NPROC (300 mg/kg/day) or IBUPROFEN (3 mg/kg/day). MIA injection produced joint degeneration including increased fissure-index, osteophyte-score, and OARSI-score, joint swelling, synovial inflammation, proteoglycan loss, and decreased cartilage thickness. These histopathological abnormalities were partially but significantly alleviated by REJENERA and other treatments. Only ZeyEX significantly inhibited the OA-induced increase in IL-1β, IL-6, IL-10 and LPO in serum, and IL-6, TNF-α, and IFN-γ in synovial fluid. While increases in MMP-3 and MMP-9 were reduced with all treatments, MMP-13 was inhibited only by REJENERA. ZeyEX increased IL-2, NPROC increased IL-6, and IBUPROFEN inhibited IL-10. All treatments improved TIMP-1 levels; however, TXNIP was more significantly inhibited by ZeyEX, and NLRP3 by REJENERA. REJENERA's anti-OA effects are accompanied by an increase in cartilage anabolic factors (Ki-67, type-II collagen, BMP-7) and inhibition of apoptosis. REJENERA offers a promising multi-targeted therapeutic approach to treating OA by blocking the iNOS-TXNIP-NLRP3 signaling axis, and reducing oxidative stress.

Gemella spp. is considered an emerging pathogen that causes endocarditis, sepsis, intracranial abscessation, and other infections in humans. A four-year-old female spayed lop-eared rabbit (Oryctolagus cuniculus) was presented for hyporexia and decreased water consumption. Physical examination revealed a lobulated right cervical mass-like lesion. Gemella cuniculi was cultured from the lesion and confirmed via whole-blood PCR and direct sequencing. CT revealed a right cervical mass involving the right salivary gland with a suspected draining tract and consolidation in the right caudal lung lobe. Antibiotics were adjusted and the mass was surgically removed. The rabbit returned to normal appetite and behavior four days post-operatively, continued systemic antibiotics for one month and remains clinically healthy today (4 mon postoperatively). Management of G. cuniculi-associated clinical disease has not been reported previously in a rabbit. G. cuniculi should be considered a possible etiologic agent for rabbit abscessation.

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 ± 1.2 mg GAE/g DW) and flavonoid (29.15 ± 0.09 mg QE/g DW) contents, exceeding those of the acetonic extract (22.2 ± 0.6 and 10.17 ± 0.07 mg GAE/g DW, respectively) and the water extract (22.4 ± 0.6 and 10.9 ± 0.6 mg QE/g DW, respectively). This extract also showed the strongest antioxidant effect, recording an IC50 of 0.060 ± 0.001 mg/mL in the DPPH assay, and an EC50 of 80 µg/mL in the RP test. In addition, it shows a great total antioxidant capacity, reaching 75.1 ± 2.0 mg EAA/g DW when compared to water and acetonic extracts (28.5 ± 1.4 and 21.1 ± 0.1 mg EAA/g DW, respectively). The antibacterial potential ranges from 0.78 ± 0.05 mg/mL to 12.6 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.

Human T-lymphotropic virus type 1 (HTLV-1) infection is associated with a broad spectrum of neurological manifestations, including conditions that may precede the development of HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). This analytical, descriptive cross-sectional study aimed to estimate the prevalence of HTLV-1 infection and describe associated sociodemographic, socioeconomic, behavioral, and clinical characteristics among patients with neurological disorders followed at a tertiary public hospital in Northeast Brazil between 2024 and 2025. A total of 300 patients underwent serological screening using an enzyme-linked immunosorbent assay (ELISA) with a commercial kit (DiaSorin Murex HTLV-1 + 2). Samples with positive or indeterminate results underwent DNA extraction and molecular confirmation by polymerase chain reaction (PCR) to differentiate viral types. Three patients tested positive for HTLV-1, resulting in an overall prevalence of 1.0%. All HTLV-1-positive individuals were women aged 43 to 72 years with low educational attainment and income. Univariate analyses identified significant associations with injection drug use, history of blood transfusion, and family history of HTLV-1 infection. Clinically, HTLV-1-positive patients presented heterogeneous neurological manifestations, ranging from urinary dysfunction and lower limb sensory and motor impairment to musculoskeletal and inflammatory symptoms. These findings highlight the broad spectrum of neurological involvement associated with HTLV-1 infection. Early identification of HTLV-1 may contribute to improved clinical management and inform public health strategies.

The OsCAMTA4 gene regulates salt and blast resistance in rice without yield loss via calcium and ABA signaling. As a key regulatory hub in the calcium signaling pathway, calmodulin-binding transcription activator (CAMTA) responds to diverse stresses and developmental signals. However, its roles in rice salt and rice blast stress responses remain largely unclear. Here, we characterized the rice CAMTA family genome-wide. Using the 3 K Rice Pan-genome and 3,000 Rice Functional Gene Haplotype Databases, we found seven core CAMTA genes are prevalent across 2,978 accessions but unevenly distributed among subgroups, with their three high-frequency haplotypes exerting distinct regulatory effects on key agronomic traits. The seven OsCAMTA genes show spatiotemporally specific responses to drought and cold stress. RT-qPCR revealed that OsCAMTA4 expression specifically was downregulated under rice blast but upregulated under salt stress. Overexpression of OsCAMTA4 enhanced salt tolerance by increasing seed germination rate, root length, proline content, and transcript levels of ABA signaling pathway genes, while decreasing malondialdehyde and hydrogen peroxide (H2O2) contents. Additionally, OsCAMTA4 knockout improved rice blast resistance by increasing proline and H2O2 accumulation and expression of disease resistance-related genes. The OsCAMTA4 protein is localized in the nucleus and interacts with OsCML2, suggesting it mediates stress responses via calcium ion (Ca2+) signaling. Notably, the actual presence of the OsCAMTA4 gene has no significant effect on rice yield over wild type, supporting its potential for improving salt tolerance and disease resistance without yield loss. Thus, it provides a new target for breeding broad-spectrum stress-resistant rice.

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.