Search Results (5,239)

Early, rapid, and accurate detection is essential for clinical management and epidemiological control of acute respiratory infections caused by pathogens. Traditional testing methods such as microbial culture, serological testing, and PCR are restrictive in terms of operation and logistics and are therefore not easily used in point-of-care settings. The CRISPR/Cas system is an adaptive prokaryotic immune system composed of clustered regularly interspaced short palindromic repeats and their associated proteins, which has been used as a nucleic acid diagnostic platform with programmable sequence-specific target recognition and signal-amplifying collateral cleavage activity. Existing reviews have mostly focused on the classification of Cas enzymes or amplification strategies; in this review, a pathogen-centric approach was taken, covering viral pathogens (SARS-CoV-2, influenza virus, RSV, HAdV and VZV), bacterial pathogens (Mycobacterium tuberculosis, Streptococcus pneumoniae, Mycoplasma pneumoniae and Staphylococcus aureus) and fungal pathogens (Aspergillus fumigatus and Pneumocystis jirovecii). Key technological advances, such as isothermal amplification coupling, single-vessel integrated reaction designs, amplification-free digital detection, and electrochemical biosensor integration, are evaluated for Cas9-, Cas12-, and Cas13-based systems, with their mechanistic bases outlined. The current challenges that hinder clinical translation, such as sample matrix interference, multiple signal cross-talk, crRNA off-target effects, and the lack of large-scale validation studies, are critically assessed. At the same time, future pathways for portable, integrated, and inexpensive diagnostic platforms are suggested. Full article

Background: Invasive aspergillosis (IA) is a severe complication after solid organ transplantation, especially in patients receiving long-term immunosuppressive therapy. Pulmonary artery involvement caused by Aspergillus fumigatus is extremely rare and may mimic pulmonary artery thrombosis or malignancy, resulting in diagnostic challenges. Case Presentation: A 61-year-old male developed fatigue, intermittent fever, cough, and chest discomfort seven months after orthotopic heart transplantation. Computed tomography pulmonary angiography revealed a mass-like filling defect involving the main pulmonary artery and left pulmonary artery, which was initially suspected to represent pulmonary artery thrombosis or tumor. Whole-blood next-generation sequencing identified Aspergillus fumigatus and Streptococcus mitis. The patient was treated with voriconazole, anticoagulation therapy, and adjustment of immunosuppressive therapy. Follow-up imaging demonstrated progressive regression of pulmonary artery lesions and disappearance of pulmonary nodules without evidence of graft rejection. Conclusions: Pulmonary artery aspergillosis should be considered in heart transplant recipients presenting with pulmonary artery-occupying lesions, particularly when imaging findings are atypical. Early pathogen identification combined with appropriate antifungal therapy, individualized immunosuppression adjustment, and careful monitoring may improve clinical outcomes. Full article

The calcium ion-transporting ATPase (Ca²⁺-ATPase) gene family maintains plant intracellular Ca²⁺ homeostasis and regulates growth, development and stress immunity; however, its functions remain poorly characterized in Malus domestica. Here, we performed a genome-wide identification of apple Ca²⁺-ATPase genes and obtained 45 members, which were classified into MdACA (39) and MdECA (6) subfamilies and unevenly distributed on 14 chromosomes. Phylogenetic analysis of Ca²⁺-ATPase genes from Malus domestica, Arabidopsis thaliana, and Oryza sativa classified these proteins into five subgroups. The ACA and ECA subfamilies were highly conserved across species, whereas Group D was apple-specific. Collinearity and Ka/Ks analyses indicated that segmental duplication and purifying selection dominated the evolution of apple Ca²⁺-ATPase genes. Promoter cis-element prediction uncovered numerous regulatory elements related to phytohormone signaling, growth, development and stress defense. Codon usage bias analysis indicated that AUG (methionine) was the dominant codon. Tissue expression profiles showed differential expression of apple Ca²⁺-ATPase genes in various organs. Quantitative real-time PCR (qRT-PCR) assays demonstrated widespread responses of Ca²⁺-ATPase genes to Alternaria alternata infection, exogenous CaCl₂, salicylic acid (SA) and methyl jasmonate (MeJA), among which MdACA39 was strongly induced under all treatments. Subcellular localization verified that MdACA39 resides on the plasma membrane. Moreover, transient overexpression of MdACA39 significantly enhanced apple resistance to A. alternata, likely due to the activation of SA, MeJA and Ca²⁺ signaling-mediated immune pathways, the induction of disease resistance-related genes, and elevated antioxidant enzyme activity. Collectively, this study systematically characterizes the apple Ca²⁺-ATPase family and identifies MdACA39 as a key regulator of fungal resistance, providing valuable gene resources for dissecting Ca²⁺ signaling-mediated disease resistance in apple. Full article

Candida infections represent a major global public health challenge and remain among the most common opportunistic fungal infections worldwide. Although Candida albicans remains the predominant pathogen, the incidence of infections caused by non-albicans species, particularly Candidozyma auris, has increased substantially in recent years. Candida infections range from superficial mucocutaneous disease to invasive life-threatening infections associated with high morbidity and mortality. In addition to clinical burden, growing evidence indicates a substantial negative impact on health-related quality of life, particularly among patients with recurrent vulvovaginal candidiasis and chronic fungal infections. Full article

Plant lipid transfer proteins (LTPs) are key components in defense against biotic and abiotic stresses, yet their functional diversity in Panax ginseng remains unclear. This study aimed to characterize LTP and evaluate its role in stress tolerance. The gene was identified from ginseng transcriptome data and analyzed using bioinformatics tools to determine its structural and physicochemical properties. Panax ginseng lipid transfer protein (PgLTP) was then heterologously expressed in Arabidopsis thaliana (A. thaliana). Transgenic lines were evaluated under fungal infection, drought, and salt stress conditions. Physiological and molecular responses, including reactive oxygen species(ROS) accumulation, malondialdehyde (MDA) content, proline levels, electrolyte leakage, and stomatal behavior under abscisic acid (ABA) treatment, were assessed. Bioinformatic analysis indicated that PgLTP encodes a small protein of approximately 12 kDa containing ten conserved cysteine residues, four α-helices, a signal peptide, and a transmembrane region, suggesting structural divergence from typical LTPs. Functional assays showed that transgenic plants exhibited significantly reduced disease indices under Fusarium oxysporum (F. oxysporum) and Cylindrocarpon destructans (C. destructans) infection. Under drought and salinity stress, transgenic lines demonstrated higher germination and survival rates, enhanced proline accumulation, reduced oxidative damage, and lower electrolyte leakage compared with the wild type. Additionally, PgLTP exhibited enhanced ABA-responsive stomatal closure, which was associated with reduced water loss. These findings indicate that PgLTP contributes to plant tolerance against both biotic and abiotic stresses, which is associated with changes in redox status, osmotic adjustment capacity, and stomatal responses. Full article

Background: Candidozyma auris (Candida auris) is an emerging multidrug-resistant fungal pathogen that can cause hospital outbreaks and is associated with high mortality. However, data on the clinical predictors of mortality in patients with bloodstream infection (BSI) caused by C. auris remain limited, particularly in Latin America, and the impact of guideline-based management practices on clinical outcomes has been scarcely explored. Methods: We conducted a multicenter retrospective cohort study including 134 patients with culture-proven C. auris BSI admitted to seven hospitals in Colombia between 2016 and 2021. Demographic, clinical, and therapeutic variables were evaluated to identify factors associated with 30-day mortality using a multivariate logistic regression model. Variables related to quality of care and guideline-based candidemia management, included in the EQUAL Candida Score, were also assessed. Results: The thirty-day mortality rate was 38.1%. In the multivariate analysis, older age (OR per decade: 1.37; 95% CI: 1.07–1.78), prior corticosteroid therapy (OR: 4.06; 95% CI: 1.70–10.16), and shock within 48 h of BSI (OR: 6.22; 95% CI: 2.52–16.52) were associated with higher mortality. Conversely, a higher EQUAL Candida Score was associated with lower 30-day mortality (OR: 0.84; 95% CI: 0.73–0.97). Conclusions: In this large Colombian cohort, advanced age, corticosteroid exposure, and early hemodynamic instability were significant predictors of mortality in patients with C. auris BSI. Greater adherence to recommended management practices—as reflected by the EQUAL Candida Score—appeared to be associated with lower mortality, underscoring the potential role of timely, structured care strategies in improving outcomes for patients with C. auris infections. To the best of our knowledge, this study is the first to evaluate the EQUAL Candida Score in a cohort of patients with C. auris BSI. Full article

Pyricularia oryzae Triticum lineage (PoTl), the fungal pathogen responsible for wheat blast disease, has demonstrated increased fitness, stable resistance to quinone outside inhibitor (QoI) fungicides, and no associated fitness cost in the absence of fungicide selection pressure. In this study, we assessed whether the fitness-related epidemiological traits and competitive abilities of QoI-resistant (R) PoTl isolates increased or remained unchanged under continuous QoI applications across multiple disease infection cycles in wheat plants, compared to untreated plants. Approximately 48 h before inoculating each group of QoI-R or sensitive (S) PoTl isolates, wheat plants were treated with 10 μg mL⁻¹ azoxystrobin. The EC₅₀, incubation and latent periods, as well as the germination capacity of the QoI-R isolates inoculated on wheat leaves treated with 10 μg mL⁻¹ azoxystrobin, remained unchanged throughout five successive infection cycles. However, continuous azoxystrobin applications throughout successive disease cycles increased head blast severity by 1.33-fold and conidial production on wheat leaves by 36.8-fold in QoI-resistant (QoI-R) PoTl isolates compared with untreated plants. Conidia germination and germ tube development by QoI-R isolates on the abaxial surface of wheat leaves and/or head rachis treated with 10 μg mL⁻¹ azoxystrobin, six hours after inoculation (h.a.i), were observed using a scanning electron microscope (SEM). These findings demonstrate that repeated azoxystrobin exposure enhances fitness-related epidemiological traits of QoI-R PoTl isolates. Consequently, alternative evolutionarily informed disease management strategies, including fungicide rotation, multisite fungicides, and integrated disease management, are urgently needed. Full article

A plant pathogen is a deterministic factor affecting the plant microbiome. However, whether the influence of different pathogen species on the plant microbiome is consistent is not well-known. In this study, mulberry leaves with similar spots caused by two diseases—mulberry brown spot (MBS, caused by Neophloeospora maculans) and mulberry snags rotten leaves disease (MSRL, caused by Boeremia exigua var. mori)—along with asymptomatic leaves, were collected. Their endophytic fungal and bacterial communities were sequenced. The occurrence of MSRL reduced leaf fungal richness and diversity (observed species from 329 to 203; Shannon diversity index from 3.6 to 2.6, p < 0.05; phylogenetic diversity index from 45.8 to 20.6, p < 0.05), along with leaf bacterial diversity; in contrast, the occurrence of MBS had little effect on both fungal and bacterial richness and diversity. Both diseases significantly altered the composition and structure of leaf fungal communities, whereas only MBS did so for bacterial communities. For the dominant genera, the fungal genera, except Cercospora, were differently affected by the occurrence of MBS and MSRL, while the bacterial genera Pseudomonas (from 8.3% to 30.6%, p < 0.05 for MBS; from 23.1% to 28.1% for MSRL), Massilia (0.2% to 7.7%, p < 0.05; 2.1% to 6.1%), Chryseobacterium (0.1% to 0.3%; 2.7% to 7%), Pantoea (2.6% to 3.7%; 0.4% to 1%), and Aureimonas (0.7% to 1.4%; 1.3% to 1.7%) were consistently enriched by both diseases. In addition, Chryseobacterium (LDA score = 4.6, p < 0.01) was detected as a biomarker after the occurrence of MSRL. Intriguingly, there were four times more significant fungi–bacteria positive associations induced by the occurrence of MSRL compared to MBS (10.2% to 2.5%). Taken together, the responses of mulberry leaf microbiome to pathogen infection vary in response to the pathogen species; although several bacterial genera were consistently enriched, whether their enrichment reflects their antagonistic roles in fungal—bacterial inter-kingdom interactions remains to be determined and should be interpreted with further experimental tests. Full article

Background and Objectives: Pediatric meningitis remains a major diagnostic and therapeutic emergency because clinical presentation may be nonspecific and early etiological distinction directly influences treatment, infection control, and epidemiological surveillance. Although bacterial meningitis has traditionally been emphasized because of its severity, the wider use of rapid molecular methods has increased the detection of viral etiologies. This study aimed to evaluate the etiological distribution of confirmed pediatric meningitis episodes and to assess temporal, age-related, and sex-associated patterns, with particular attention to the observed shift from bacterial toward viral meningitis. Materials and Methods: A retrospective single-center analysis was performed using microbiological data from suspected meningitis episodes investigated between 2023 and 2025 at the “Louis Țurcanu” Emergency Hospital for Children, Timișoara. Confirmed episodes were classified as bacterial, viral, or fungal according to the identified etiological agent. The distribution of suspected and confirmed episodes, annual etiological patterns, age-group differences, sex-associated variation, and identified pathogens were assessed using descriptive statistics and trend analysis. Results: Among 641 suspected episodes, 58 were microbiologically confirmed, corresponding to an overall positivity rate of 9.05%. Viral meningitis was the most frequent etiology, accounting for 30 episodes (51.72%), followed by bacterial meningitis with 22 episodes (37.93%) and fungal meningitis with 6 episodes (10.34%). Etiological distribution changed significantly over the study period: viral episodes increased from 22.22% in 2023 to 77.78% in 2025, while bacterial episodes decreased from 66.67% to 22.22%. Viral meningitis predominated in children aged 1–6 years and was more frequent among male patients, whereas bacterial meningitis remained more frequent in infants. Enterovirus became the dominant identified agent in 2025. Conclusions: The local epidemiology of confirmed pediatric meningitis showed an observed redistribution among confirmed episodes toward viral etiologies, particularly enteroviral meningitis, alongside a proportional decline in bacterial episodes. These findings support continued etiological surveillance and the integration of rapid molecular testing with conventional microbiology in the diagnostic management of suspected pediatric meningitis. Full article

Fungal infections remain a major global health challenge, underscoring the need to better understand host–pathogen interactions. Extracellular vesicles (EVs) released by fungal pathogens are emerging as key modulators of immune responses. Here, we investigated how EVs from Candida albicans and Aspergillus fumigatus influence macrophage activation, focusing on the integration of pattern recognition receptors (PRRs) and cytosolic sensing pathways. EVs were characterized by nanoparticle tracking analysis, zeta potential, and cryo-electron microscopy. Human THP-1-derived macrophages were stimulated with fungal EVs, and cytokine production, gene expression, and signaling pathway activation were assessed by ELISA, RT-qPCR, and Western blotting. C. albicans EVs induced a strong pro-inflammatory response, with increased production of IL-1β, IL-6, TNF, IL-8, and IFN-β after 24 h. In contrast, A. fumigatus EVs elicited a more limited response, characterized by IL-4 production and minimal pro-inflammatory cytokine induction. Both EV populations modulated PRR expression and activated the cGAS–STING–IRF3 axis; however, C. albicans EVs promoted coordinated activation of TLR4, TLR9, and Dectin-1–CARD9 signaling, whereas A. fumigatus EVs downregulated CARD9 and dampened inflammation. These findings demonstrate that fungal EVs differentially shape macrophage responses through species-specific integration of immune sensing pathways. Full article

Background: Saffron corm rot causes significant yield losses worldwide. Although synthetic pesticides are commonly used for management, their overuse can lead to increased fungal resistance and pose a risk to public health. Numerous plants possessing antifungal properties hold significant potential for development as biocontrol agents against saffron corm rot. Result: In this study, the efficacy of biocontrol agents derived from natural plants was evaluated for managing saffron corm rot. Aqueous and 70% ethanol extracts from 15 plants were investigated for antifungal activity and virulence suppression against three saffron pathogens: Fusarium oxysporum, Penicillium citrinum, and Aspergillus brasiliensis. Antifungal activity experiments indicated that clove ethanol extract at a concentration of 20 mg/mL exhibited the highest inhibition rate, with values of 93.27 ± 0.0% for Fusarium oxysporum, 86.47 ± 5.55% for Penicillium citrinum, and 68.65 ± 2.40% for Aspergillus brasiliensis. Compared with the model group, clove ethanol extract significantly reduced corm rot and normalized the metabolism of infected corms. On day 30, compared with the model group (34.3 ± 3.7%), the rot rate of clove-treated corms was reduced to 17.2 ± 1.4%, and on day 60, it was reduced to 27.5 ± 1.5% (versus 52.9 ± 3.3% in the model group). Compared with the model group, the levels of soluble sugar, starch, α-amylase, soluble protein, and superoxide dismutase in clove-treated infected corms approached those observed in the control group. Field trials confirmed that clove ethanol extract effectively improved plant growth and reduced the disease severity index (37.00 ± 3.51) in infected corms. Conclusion: Clove ethanol extract is a promising candidate to replace or reduce the use of synthetic pesticides. It provides a theoretical and practical basis for the biocontrol of saffron corm rot. Full article

Amomum villosum is an evergreen perennial herb widely distributed in tropical and subtropical regions, with significant economic and medicinal importance. Throughout the cultivation process, it faces severe fungal disease problems that significantly impact its yield and quality. In July 2025, leaf spot disease with a 56% incidence was observed on A. villosum in Wenshan, Yunnan, China. Affected leaves initially developed irregular grayish-white lesions surrounded by brown margins. As symptom development progressed, semi-submerged black spots formed on the lesion surfaces. Severe infections resulted in premature leaf abscission and, ultimately, death of the entire plant. To identify the pathogen responsible, we conducted isolation and pathogenicity studies. Through morphological characterization, phylogenetic analysis (ITS, LSU, and TUB), and pathogenicity tests, Pestalotiopsis microspora was determined as a pathogen. Koch’s postulates were fulfilled on attached leaves. After 15 days, typical necrotic lesions appeared on inoculated leaves, while controls remained symptom-free. This is the first report of A. villosum leaf spot caused by P. microspora in China. Biocontrol assays revealed that Trichoderma harzianum T15 and T. asperellum T16 exhibited significant antagonistic activity against P. microspora, with inhibition rates of 44.77% and 50.70%, respectively. In addition, Bacillus velezensis SWFU41, isolated from healthy A. villosum leaves, showed superior disease suppression, achieving a control efficacy of 60.52%. Compared with the fungal biocontrol agents, B. velezensis demonstrated greater inhibitory activity against the pathogen, suggesting that bacterial antagonists may provide a more effective biological control strategy for managing A. villosum leaf spot disease. This is the first report of P. microspora-caused leaf spot disease on A. villosum. Furthermore, the comparative evaluation of fungal and bacterial antagonists demonstrated that B. velezensis SWFU41 outperformed T. harzianum and T. asperellum in disease suppression, highlighting its potential as a promising biocontrol agent for sustainable disease management. These findings provide a scientific basis for pathogen monitoring, epidemiological studies, and the development of integrated biological control strategies for A. villosum cultivation. Full article

Invasive fungal infections, including candidemia and invasive aspergillosis, remain significant causes of morbidity and mortality in at-risk patient populations. Rapid, accurate diagnosis is critical for timely antifungal therapy and improved clinical outcomes. We evaluated two commercial molecular assays, the CandID Multiplex PCR and AspID Real-Time PCR, for detection of clinically relevant Candida and Aspergillus species, respectively, in at-risk patients in Paraguay. The CandID assay demonstrated strong agreement with blood culture in detecting candidemia. The AspID assay showed high concordance with galactomannan testing, particularly in bronchoalveolar lavage specimens, supporting its role as a complementary diagnostic tool. Our findings underscore the value of integrating molecular diagnostics into routine fungal diagnostic workflows to improve early detection of invasive fungal infections. Full article

Indoor-air research has largely examined infection, microbial ecology, immune effects and antimicrobial resistance separately, leaving the pathway from indoor biological sources to chronic respiratory outcomes insufficiently integrated. This narrative review synthesises evidence on indoor airborne pathogens and non-viable microbial components as health-relevant biological exposures beyond acute infection. Literature published between 2000 and February 2026 was reviewed from PubMed, Scopus and Web of Science, supplemented by guidance from WHO, ECDC, US EPA and ASHRAE. Viable microorganisms and non-viable components, including endotoxin, β-(1→3)-glucans, microbial DNA and extracellular vesicles, engage epithelial pattern-recognition pathways and promote inflammatory signalling. Findings included 6.5% higher TNF-α and 5% higher IL-8 per log-unit increase in fungal-spore exposure among sawmill workers; uncontrolled asthma in 45% of moisture- or mould-exposed versus 33% of non-exposed children; airborne resistance-gene and mobile-element loads of 0.55–479.44 copies/m³ in hospital departments; and a 32.8% reduction in viral diversity, but no significant reduction in high viral exposure, following classroom HEPA filtration. These findings support biological plausibility but reveal a fragmented evidence base dominated by observational studies, heterogeneous sampling and limited longitudinal exposure–response data. Indoor bioaerosols should be considered continuous exposures within the exposome, requiring research and regulation across microbiology, environmental engineering, medicine and public health. Full article

Fragrant pear canker, caused by Cytospora pyri, is a major branch disease that reduces the productivity and longevity of Korla fragrant pear orchards in Xinjiang, China. During infection and lesion expansion in woody tissues, C. pyri must breach the host cell wall barrier, in which pectin degradation plays a central role by weakening intercellular adhesion and disrupting tissue integrity. Members of glycoside hydrolase family 28 (GH28), represented mainly by polygalacturonases and other pectin-degrading enzymes, are closely involved in fungal invasion and colonization. However, the composition and virulence-related functions of this gene family encoding for these enzymes in C. pyri remain unclear. In this study, GH28 genes were systematically identified and comparatively analyzed in C. pyri and closely related Cytospora species. Infection-stage expression profiling and functional validation were then performed to assess their roles in pathogenicity. A total of 73 GH28 genes were identified across five Cytospora species, including 15 in C. pyri. Most C. pyri GH28 proteins were predicted to be acidic, hydrophilic, extracellularly secreted proteins carrying conserved motifs. Phylogenetic analysis showed that C. pyri GH28 members were closely related to homologs from C. mali. Genomic distribution analysis revealed that these genes were dispersed across multiple scaffolds, with no obvious tandem duplication events. RT-qPCR analysis showed that all seven candidate GH28 genes were induced during infection of fragrant pear branches, with VP1G_08835 and VP1G_03209 exhibiting strong expression responses at 6 dpi. Functional validation further showed that deletion of VP1G_08835 impaired vegetative growth and reduced lesion length on detached pear branches by 26.62% compared with the wild type, whereas complementation restored these phenotypes. These findings demonstrate that GH28 genes participate in C. pyri infection and identify VP1G_08835 as an important GH28 member required for normal growth and contributing to virulence. Full article