Oysters have high nutritional value; however, their short shelf life limits their commercialisation to areas close to production sites. Affordable and accessible processing techniques that extend shelf life could expand both market reach and consumer access to oysters. This study evaluates the physicochemical, microbiological and sensory properties of raw Crassostrea gigas oyster meat semi-preserved with weak organic acids and saline solutions under refrigerated storage (4 °C). Aliquots of (100±2) g of raw oyster meat were placed into plastic containers containing different solutions: sterile deionised water only (negative control treatment, labelled W), base solution (NaCl, 5 % m/V) only, base solution with 2 % citric acid (CA), base solution with 2 % lactic acid (LA),and base solution with 2 % acetic acid (AA). Sensory, physicochemical and microbiological characteristics of the semi-preserves were monitored for 16 days. Incorporating weak acids into the marination solutions effectively inhibited the growth of mesophilic and psychrotrophic bacteria in semi-preserved oysters during 16 days of refrigerated storage. In contrast, water and NaCl treatments exceeded the recommended limit of 5 log CFU/g for seafood after 3 and 11 days, respectively. By day 16, total volatile basic nitrogen (TVB-N) values indicated early spoilage in water ((30.0±1.1) mg/100 g), satisfactory freshness in NaCl ((24.5±2.6) mg/100 g) and CA ((18.3±1.1) mg/100 g), and excellent freshness in LA ((14.7±0.0) mg/100 g) and AA ((15.0±1.8) mg/100 g). Thiobarbituric acid reactive substances (TBARS) assay values, expressed as malondialdehyde (MDA), remained below 3 mg/kg in all treatments, indicating good oxidative stability. Among the acids, AA maintained higher pH values (3.78 on day 16) than CA (3.26) and LA (3.14) and showed the lowest microbial loads; however, it received the highest scores for acid odour (median=5.35) and the lowest for characteristic oyster odour. CA and LA produced sensory profiles more similar to fresh oysters, with higher characteristic odour scores and lower acid odour scores, but slightly higher spoiled odour scores (still low in absolute terms). Overall, AA was the most effective for microbiological and physicochemical preservation, while CA and LA offered better sensory acceptance. These results highlight the potential of weak organic acids, particularly AA, as a low-cost method to extend the shelf life of raw oysters to at least 16 days under refrigeration. This study evaluates the effects of marination with weak acids on the physicochemical, microbiological, and sensory properties of raw oyster meat. The wide range of parameters analysed highlights not only its suitability for consumption but also consumer preferences based on sensory aspects such as colour and odour. The combined findings can assist the industry in selecting the most appropriate acid for developing different oyster-based products.
Late-onset sepsis (LOS) remains a leading cause of morbidity and mortality in preterm infants, with decreasing incidence in high-income countries but persistent challenges in low- and middle-income countries. This study aimed to evaluate the epidemiology, microbiological profile, and short-term outcomes of LOS in very low birth weight (VLBW) preterm infants in Brazil. A multicenter cohort study across 18 level 3 neonatal units of the Brazilian Network on Neonatal Research included 13,439 VLBW infants (400-1499 g birth weight, 22-36 weeks gestation) admitted between 2010 and 2020, excluding those with major malformations, congenital infections or death before 72 hours. Main outcomes included incidence of proven and clinical LOS, in-hospital mortality, and associated short-term morbidities. The cohort had a mean gestational age of 29 ± 3 weeks and birth weight of 1079 ± 275 g. The incidence of proven LOS was 24.6%, and clinical LOS was 19.2%. Gram-positive bacteria predominated (64.1%, with 49.4% coagulase-negative staphylococci), followed by Gram-negative bacteria (27.2%) and fungi (8.8%). Over the decade, fungal infections decreased, while Gram-negative bacterial infections and proven LOS incidence increased. In-hospital mortality was 24.8%, showing no reduction. LOS was associated with an increased risk of death and severe morbidities. This comprehensive 10-year cohort in Brazil reveals a concerning epidemiological shift, characterized by a significant increase in Gram-negative infections and a persistent, high LOS incidence and mortality among VLBW infants. These findings, particularly the lack of improvement in prognosis, underscore the urgent need for targeted and effective preventive and control interventions tailored for resource-limited settings.
This study evaluated the stability of extemporaneous pyrimethamine oral suspensions prepared from commercial tablets using 85% sucrose or 70% sorbitol as vehicles. A validated HPLC method was employed to quantify pyrimethamine and assess its chemical stability. Physicochemical parameters including macroscopic appearance, pH, particle size, and drug content were monitored for 90 days at 5 and 25 °C. Both formulations maintained acceptable pH values and pyrimethamine content within pharmacopeial limits. Particle size increased over time, likely due to aggregation, while appearance remained unchanged for up to 45 days. Microbiological assessments showed that microorganisms remained within acceptable limits, with no objectionable microorganisms detected. Overall, both suspensions demonstrated chemical and microbiological stability for up to 45 days (Suspension S) and 60 days (Suspension X) under both storage conditions. These results support the preparation of cost-effective pyrimethamine formulations for treating congenital toxoplasmosis in pediatric patients.
Gestational diabetes mellitus (GDM), characterized by pregnancy-induced insulin resistance, and periodontal disease, a chronic inflammatory condition of the periodontium, are intricately linked and may collectively amplify the risk of adverse maternal and fetal outcomes. This study aimed to evaluate periodontal microbial associations in periodontitis in relation to GDM. An observational cross-sectional study was conducted among 80 pregnant women with periodontitis, stratified into GDM (Group A; n = 40) and non-GDM (Group B; n = 40) groups. Clinical parameters, including Plaque Index, Oral Hygiene Index-Simplified, Modified Sulcular Bleeding Index, and Probing Pocket Depth (PPD), along with glycemic parameters (OGTT), were recorded. Subgingival plaque samples were analyzed using real-time Polymerase Chain Reaction for key periodontal pathogens (Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Selenomonas sputigena, Treponema denticola, and Tannerella forsythia). Pearson correlation analysis assessed associations between clinical and glycemic parameters and pathogens, followed by multiple linear regression to determine the independent effects of predictors on the outcomes (p < 0.05). Group A demonstrated significantly lower Ct values (higher bacterial load) for P. gingivalis, S. sputigena, P. nigrescens, T. denticola, and F. nucleatum (p ≤ 0.005). Strong positive correlations in the overall population were observed between P. gingivalis and probing pocket depth (r = 0.79, p = 0.001) and clinical attachment loss (r = 0.73, p = 0.010), followed by S. sputigena (r = 0.64, p = 0.01). Multiple regression analysis identified MSBI, PPD, and CAL as significant predictors of P. intermedia and P. gingivalis in both groups (p < 0.05), with OGTT-1h emerging as an additional predictor of P. gingivalis. In contrast, F. nucleatum and S. sputigena demonstrated significant inverse associations with these parameters (p < 0.05). The results of the present study demonstrate that GDM is associated with an increased periodontal pathogenic burden, with clinical and glycemic parameters emerging as significant independent determinants, highlighting the intricate interplay between metabolic dysregulation and periodontal disease.
SUMMARYPlasmodium vivax remains a major barrier to global malaria elimination due to dormant liver-stage hypnozoites that drive relapse and sustain transmission even in low-endemic settings. India accounts for nearly half of the global P. vivax burden, placing it at the center of elimination efforts. This review synthesizes current evidence on the epidemiology, biology, and control of P. vivax in India. Despite progress, early gametocytogenesis, hypnozoites, low parasitemia, and immune evasion complicate diagnosis and treatment. Recent single-cell and multi-omics studies are elucidating vir gene regulation, epigenetic control of hypnozoite dormancy, and quiescent liver-stage biology. Diagnostic gaps persist because microscopy and rapid diagnostic tests poorly detect asymptomatic infections, and molecular surveillance cannot map relapse without hypnozoite biomarkers. Standard radical cure with chloroquine and 14-day primaquine is limited by poor adherence and restricted glucose-6-phosphate dehydrogenase testing, while single-dose tafenoquine awaits wider implementation. Urban malaria, largely driven by P. vivax and Anopheles stephensi, is increasing as a consequence of rapid urbanization and vector adaptation. National initiatives such as the Integrated Health Information Platform, Tribal Malaria Action Plans, and Urban Malaria Schemes are critical but face constraints in surveillance integration, private sector reporting, and cross-border coordination. We highlight emerging experimental liver models, single-cell technologies, and systems biology frameworks as essential tools for addressing key knowledge gaps in relapse biology. Sustained investment and interdisciplinary collaboration will be required for India to lead P. vivax elimination and translate national progress into global impact.
Cryptococcosis is a severe invasive fungal infection with limited therapeutic options beyond fluconazole-based regimens. Isavuconazole, a broad-spectrum triazole antifungal, has emerged as a potential alternative, although clinical data supporting its use remain scarce. We aimed to evaluate the real-world effectiveness and safety of isavuconazole in patients with different forms of cryptococcosis. A retrospective observational study was conducted at a tertiary-care hospital, including patients with cryptococcosis who received isavuconazole at any treatment phase. Standard microbiological methods were used for pathogen identification and susceptibility testing. Demographic, clinical, and microbiological data were collected. Clinical and microbiological responses and tolerability were assessed at end of treatment or until death. Eight patients with cryptococcosis received isavuconazole, most of whom were immunocompromised. Clinical presentations included pulmonary and disseminated disease, with Cryptococcus neoformans as the predominant species. Isavuconazole was primarily used during the consolidation and maintenance phases, after induction therapy with amphotericin B and flucytosine for 2 weeks in most cases, and as salvage therapy in two patients. It was well tolerated during prolonged treatment (6-12 months). In the two patients with isavuconazole therapeutic drug monitoring, plasma total trough concentrations were within the therapeutic range (5 and 3.5 µg/mL, respectively), whereas cerebrospinal fluid total concentration levels were undetectable. A favorable clinical response was observed in four patients, while three remain on treatment with ongoing clinical improvement; one patient died early. Microbiological clearance was achieved in all culture-positive cases. Isavuconazole demonstrated clinical effectiveness in this cohort of patients across different presentations of cryptococcosis. Treatment was safe and well tolerated, supporting its role as an alternative antifungal option against Cryptococcus, particularly when fluconazole is limited by adverse effects or drug-drug interactions. However, data on central nervous system penetration were limited, and further studies are needed to better define its role in cryptococcal meningitis management.
To develop predictive models for early and overall tuberculosis (TB) deaths for prospective use at TB diagnosis in resource-constrained TB programme settings. Statewide cohort study using routinely captured secondary data. With the majority of TB deaths being early (within 2 months), India's TB programme's information management system (Ni-kshay)-dependent death prediction models (using age, gender, TB site, previous treatment, microbiological confirmation, HIV, diabetes and bank account availability) are not feasible for prospective use, as few variables are captured at diagnosis. Utilising routinely captured triage variables for severe illness at diagnosis (body mass index, pedal oedema, respiratory rate, oxygen saturation and ability to stand without support) from an ongoing statewide and state-specific differentiated TB care initiative to reduce TB deaths in Tamil Nadu state (southern India, 80 million population with 0.1 million annual notifications), robust models for prospective use were developed. Adults (aged ≥15 years) with TB (not known to be drug-resistant at diagnosis) that were notified from public facilities of Tamil Nadu from July 2022 to June 2023. Early and overall (within 12 months of notification) TB deaths. Area under the receiver operating characteristic curve (AUC) was used to assess accuracy of models built using modified Poisson regression. Among 55 971 adults, the overall death rate was 7.4%, and 67.9% of the deaths were early. In predicting overall deaths, accuracy of the model using all Ni-kshay variables (AUC 0.716 (95% CI 0.707 to 0.725)) was as good as the model using triage variables for severe illness only (AUC 0.701 (95% CI 0.691 to 0.711)). To the latter, adding potentially capturable Ni-kshay variables at diagnosis (age, gender, TB site, previous treatment and microbiological confirmation) significantly improved model accuracy (AUC 0.754 (95% CI 0.745 to 0.763)). Further addition of remaining Ni-kshay variables did not improve accuracy significantly. Death prediction equations were generated for these models. Simple and easily measurable triage variables for severe illness should be routinely captured at TB diagnosis. A death prediction calculator (http://44.208.93.99/) based on these variables (specifically triage variables for severe illness combined with age, gender, TB site, previous treatment and microbiological confirmation) may be used by Indian states and high TB burden countries seeking scalable, data-driven interventions to reduce TB deaths.
To evaluate the diagnostic performance of nanopore-targeted sequencing (NTS) for sputum smear-negative pulmonary tuberculosis (PTB). In this retrospective study, bronchoalveolar lavage fluid from patients with suspected PTB and negative sputum smears was tested with NTS, Xpert MTB/RIF, and MGIT 960 culture at Shanghai Pulmonary Hospital (August 2022-March 2024). Sensitivity and specificity were evaluated against both microbiological and composite reference standards. Receiver operating characteristic curves were generated and Youden's index was employed to determine the optimal NTS cutoff. Among the 249 participants enrolled, 100 were microbiologically confirmed PTB, 33 were probable PTB, and 116 were tuberculosis-negative. Under microbiological reference standard, the sensitivity of NTS was 0.940 (95% confidence interval [CI], 0.869-0.975), higher than Xpert MTB/RIF (0.840 [95% CI, 0.750-0.903]; P = .041) and MGIT 960 culture (0.640 [95% CI, 0.537-0.732]; both P < .001). The specificity of NTS was 0.776 (95% CI 0.687-0.846), compared to 0.974 (95% CI 0.921-0.993) for Xpert MTB/RIF and 1.000 (95% CI 0.960-1.000) for MGIT 960 culture. Excluding patients with prior PTB history, NTS specificity increased to 0.856 (95% CI, 0.766-0.916). According to Youden's index, when 4 Mycobacterium tuberculosis-specific sequence reads was defined as the optimal NTS cutoff for positive NTS result, the specificity of NTS rose to 0.907 (95% CI, 0.827-0.954). NTS offers rapid, highly sensitive detection of PTB in patients with low bacillary burden. Applying an optimal read-count cutoff and accounting for prior PTB history can enhance clinical utility.
Tuberculosis and nontuberculous mycobacterial pulmonary disease are major causes of chronic respiratory morbidity, yet detailed multicenter clinical data remain limited. We established a retrospective multicenter cohort using electronic medical records from 4 national university-affiliated tertiary hospitals in Korea. Eligible participants included patients with tuberculosis registered in the Korean National Tuberculosis Surveillance System and patients who met modified 2007 American Thoracic Society/Infectious Diseases Society of America microbiological criteria for nontuberculous mycobacterial pulmonary disease between 2015 and 2024. De-identified, standardized datasets were created using common data specifications and harmonized at a central coordinating center. The cohort included 11,790 patients with tuberculosis and 5,145 patients with nontuberculous mycobacterial pulmonary disease. Tuberculosis notifications declined steadily from 1,657 cases in 2015 to 580 cases in 2024, whereas nontuberculous mycobacterial pulmonary disease showed an overall upward trend despite a modest decrease in the final year. Mycobacterium avium complex accounted for 78.7% of nontuberculous mycobacterial pulmonary disease cases, with M. intracellulare and M. avium as the predominant species. Macrolide-based multidrug regimens were used consistently throughout the study period. This cohort profile describes the registry design, case definitions, data structure, and key baseline findings; the registry provides a platform for future clinical, epidemiological, and health-services research on mycobacterial disease in Korea.
Extended-spectrum β-lactamase (ESBL)-producing Enterobacterales are an increasing cause of complicated urinary tract infections (UTIs). Reliance on carbapenems to treat these infections accelerates resistance, underscoring the urgent need for effective oral transition options to preserve last-line agents and lessen healthcare burden. We conducted a multicenter, open-label, randomized, noninferiority trial at four tertiary hospitals in South Korea (November 2022-June 2025). Adults hospitalized with complicated UTIs due to ESBL-producing Enterobacterales who improved after 3-7 days of intravenous antibiotics were randomized (1:1) to either continue intravenous carbapenem or β-lactam/β-lactamase inhibitor therapy or switch to oral fosfomycin trometamol (3 g once daily). The primary outcome was clinical cure, defined as resolution of urinary tract infection-related symptoms and signs within 4 days after completion of treatment. Secondary outcomes included microbiological cure, readmission, adverse events, and 30-day recurrence. Of the 344 screened patients, 299 were randomized (152 oral fosfomycin; 147 intravenous therapy). Clinical cure occurred in 92.8% of the oral fosfomycin group and 95.2% of the intravenous group (risk difference, -2.47%; 95% CI -7.84 to 2.89), confirming noninferiority. Microbiological cure was similarly high in both groups: urine (98.0% vs 96.6%) and blood (97.3% vs 97.5%). Safety profiles and 30-day outcomes were also comparable. Oral fosfomycin was noninferior to continued intravenous therapy when used as oral transition therapy for complicated UTIs caused by ESBL-producing Enterobacterales. These findings provide evidence to support oral fosfomycin transition as a carbapenem-sparing antibiotic stewardship strategy for patients with complicated UTIs caused by ESBL-producing Enterobacterales. KCT0007669.
Conventional microbiological tests have limitations in the microbial diagnosis of immunocompromised patients. Next-generation sequencing (NGS) technologies have the potential to overcome some of these challenges by enabling rapid, comprehensive, and hypothesis-free pathogen detection, potentially improving the speed and accuracy of microbial diagnosis and subsequent clinical outcomes. This review summarizes current evidence for the use of NGS technologies in immunocompromised populations, highlights areas of demonstrated clinical impact, and identifies key priorities for broader clinical integration. Case reports and series have demonstrated the utility of NGS in diagnosing unusual or atypical infections amongst immunocompromised patients that were initially missed by conventional methods. Retrospective observational studies indicate that NGS can achieve higher sensitivity and greater pathogen detection rates than conventional diagnostics, although performance may be limited for certain pathogens, such as Aspergillus and Mycobacterial species. The clinical impact of NGS-guided interventions varies, reflecting both differences in study design and challenges in interpreting metagenomic data. NGS technologies have the potential to enhance microbial diagnosis in immunocompromised patients, particularly in complex, polymicrobial, or atypical infections where conventional methods fail. However, widespread clinical adoption is limited by high costs, complex workflows, and the need for advanced bioinformatics infrastructure and expertise. Further research is required to define clinical impact, cost-effectiveness, and to standardize workflows and guide optimal time for implementation, in order to inform evidence-based integration of NGS into routine clinical practice.
This study aimed to investigate the fermentation of reduced-calorie sour cherry-based beverages using three individual probiotic strains (Lactiplantibacillus plantarum SH5, Limosilactobacillus fermentum SH10, and Lactiplantibacillus pentosus SH14) and one mixed-strain formulation, and to evaluate these beverages together with unfermented control beverages in terms of their physicochemical, microbiological, bioactive, and cytotoxic properties during 28 days of cold storage. The fermented beverages exhibited pH values of 3.54-3.63, turbidity of 0.614-0.779, total soluble solids of 24.93-29.87 °Brix, and dry matter content of 27.62%-30.75%, with color parameters recorded as L* (15.08-16.66), a* (12.80-18.21), and b* (-5.78 to -3.05). Phenolic profiling identified catechin (31.05-32.90 ppm), chlorogenic acid (10.82-11.05 ppm), and myricetin (5.05-6.90 ppm) as the predominant compounds, although most phenolics gradually declined during storage. Despite higher initial total phenolic content (TPC) in control samples, lacto-fermented beverages exhibited superior recovery values (%), indicating enhanced bioaccessibility during in vitro digestion. Calorie reductions reached up to 46.46% in SCN9-SH10, while viable LAB populations remained above 106 CFU/mL throughout 21 days of storage (7.01-7.65 log CFU/mL). TPC ranged from 214.75 to 353.67 mg gallic acid equivalent (GAE)/100 mL, DPPH scavenging activity from 213.54 to 274.02 mg Trolox equivalent (TE)/100 mL, and cupric reducing antioxidant capacity (CUPRAC) values from 924.46 to 1576.71 mg TE/100 mL, with fermented samples consistently demonstrating higher antioxidant capacity than controls. Antibacterial activity was observed only in SCN9-Mix and control samples against Bacillus cereus ATCC 11778, while cytotoxicity assays indicated modest inhibitory effects toward MCF-7 breast cancer cells, with cell viability ranging from 87.64% to 94.45% in freshly prepared samples and decreasing over 28 days. Collectively, these findings demonstrate that lactic acid fermentation effectively enhances the functional, microbiological, and bioactive properties of reduced-calorie sour cherry-based beverages.
Tunneled central venous catheters (CVC) are essential vascular access for many maintenance hemodialysis(HD) patients, yet catheter-related bloodstream infections (CRBSI) remain a persistent complication driven by biofilm formation and multidrug-resistant organisms, creating a clinical dilemma between source control (catheter removal) and vascular access preservation. This narrative review synthesizes current evidence across HD-CRBSI epidemiology, pathogenesis, prevention, treatment, and future directions, and provides an evidence-based, decision-oriented management framework. Key contributions include: systematic distinction between extraluminal (skin → cuff → bloodstream) and intraluminal (hub contamination) routes of infection; an HD-specific pathogen profile establishing Gram-positive predominance (approximately 64% of cases, with coagulase-negative staphylococci accounting for approximately 32%); a structured clinical classification differentiating uncomplicated versus complicated CRBSI and exit-site, tunnel, and systemic infections; explicit criteria for catheter salvage (hemodynamically stable patients with uncomplicated CRBSI caused by low-virulence pathogens, mandatory clinical and microbiological reassessment at 48-72 hours, and documented clearance of bacteremia) versus mandatory catheter removal (Staphylococcus aureus, Pseudomonas aeruginosa, Candida spp., tunnel infection, septic shock, or persistent bacteremia >72 hours); systematic integration of antimicrobial lock therapy into the salvage pathway, supported by HD-specific evidence including the LOCK-IT-100 trial; prioritization of standard-of-care prevention practices (mandatory catheter hub disinfection, standardized aseptic connection/disconnection protocols, staff training with care bundles) over experimental strategies; and identification of key research priorities including standardized CRBSI surveillance methodologies, prospective validation of AI-assisted diagnostic tools, and rigorous evaluation of next-generation preventive technologies.
Rapid, accurate, and on-site detection of foodborne pathogens remains a critical challenge in ensuring food safety. Traditional microbiological methods, including culture-based assays, ELISA, and PCR, often require long processing times, specialized equipment, and trained personnel, limiting their practical application in real-time monitoring. Electrochemical biosensors have emerged as promising alternatives due to their portability, fast response, low cost, and compatibility with complex food matrices. Molecularly imprinted polymers (MIPs) offer a robust synthetic recognition layer for these sensors, providing high chemical stability and tenable selectivity while overcoming limitations of biological recognition elements such as antibodies and enzymes. However, conventional MIP fabrication often relies on empirical trial-and-error approaches, which can reduce reproducibility and slow sensor development. This review summarizes recent advances in computationally guided MIP design for electrochemical biosensing of key foodborne pathogens, including Salmonella spp., Listeria monocytogenes, Escherichia coli O157:H7, Staphylococcus aureus, and Pseudomonas aeruginosa. We discuss how molecular docking, density functional theory, and molecular dynamics simulations can predict template-monomer interactions, optimize cavity formation, and guide polymer properties, thereby improving sensor sensitivity, selectivity, stability, and response time. By integrating computational modeling with experimental electrochemical methods, these approaches enable a mechanism-driven development of portable, high-performance biosensors for microbial detection. This convergence of computational and microbiological methods provides a pathway toward next-generation, ready-to-use devices for rapid foodborne pathogen monitoring.
Listeriosis remains one of the most severe foodborne diseases in terms of fatality rate. Listeria monocytogenes can grow under stressful conditions and contaminate various food categories. Regulation (EC) No 2073/2005 modified on microbiological criteria for foodstuffs includes both qualitative and quantitative food safety criteria on L. monocytogenes and mentions EN ISO 11290-2 Standard as the reference enumeration method for the quantitative criteria. As food products are usually contaminated at very low levels, more sensitive enumeration methods are necessary to provide reliable data for research studies or routine analysis. This study, conducted by the EURL Lm, in collaboration with a group of NRLs, allowed to evaluate 2 more sensitive alternative methods derived from ISO 11290-2 Standard. In particular, the practicability, relative trueness, repeatability and accuracy profile were examined.
Non-sputum-based tools are essential complements to the current tuberculosis diagnosis strategy. We aim to determine whether dual-gene qPCR melting curve detection of salivary exosomes Mycobacterium tuberculosis DNA can diagnose paucibacillary tuberculosis. We developed the exosomal nucleic acid (ExoNA)-based saliva tuberculosis diagnostic, a rapid, non-sputum diagnostic method for tuberculosis by integrating enrichment of salivary exosomes, nucleic acid extraction, and dual-target qPCR melting curve analysis. In a pilot clinical study, adults with suspected pulmonary tuberculosis were enrolled to evaluate diagnostic performance against multiple reference standards. Subgroup analysis was performed in patients with negative smear microscopy. Salivary exosomal DNA was identified as the predominant form of detectable Mycobacterium tuberculosis DNA in saliva. ExoNA-based salivary tuberculosis diagnostic reduced the turnaround time from 16 h to 3h, and reached a limit of detection of 10 CFU/mL. In total, 130 patients with suspected pulmonary tuberculosis were included. Of them, 36·9% (48/130) were asymptomatic, and 80·8% (105/130) had negative smear microscopy results. The ExoNA-based Saliva TB assay had sensitivity of 81.2% (95% CI 53.7%-95.0%) and specificity of 82.8% (95% CI 63.5%-93.5%) against microbiological reference standards (n=45), while sensitivity of 68.4% (95% CI 54.6%-79.7%) and specificity of 82.9% (95% CI 67.4%-92.3%) under extended microbiological reference standards (n=98). In the subset of 105 patients with negative smear microscopy, the sensitivities remained within the range of 67.5% to 75.0% across different reference standards, and specificities ranged from 82.1% to 82.9%. ExoNA-based saliva tuberculosis assay demonstrates favorable diagnostic performance in patients with suspected pulmonary tuberculosis, with the advantages of non-invasive nature, easy sampling, and rapid detection. Multicenter validation in more heterogeneous populations is necessary prior to clinical use.
Optimization of urine lipoarabinomannan (LAM) detection platforms holds significant potential for improving early tuberculosis (TB) diagnosis. Currently, accurate urine LAM assays are notably lacking for HIV-negative populations. To address this gap, we evaluated the diagnostic accuracy of an Ultra-sensitive electrochemiluminescence LAM assay (AIMLAM) in HIV-negative individuals with suspected pulmonary TB in Beijing, China. In this prospective multicentre study, we enrolled adult (≥ 18 years), presumptive TB patients in Beijing, China. All participants provided sputum samples for smear microscopy, Xpert MTB/RIF assay, and mycobacterial culture, as well as urine samples for AIMLAM testing. Diagnostic performance was evaluated against both microbiological reference standard (MRS) and a composite reference standard (CRS). We collected and stored urine samples from 692 HIV-negative adult presumptive TB patients. Of these, 315 were microbiologically confirmed TB cases, 177 had possible TB, and 200 were considered unlikely to have TB. When evaluated against the MRS, the AIMLAM assay demonstrated a sensitivity of 61.6% (95% CI: 56.1-66.8) and specificity of 83.0% (95% CI: 78.9-86.5). Using the CRS, the AIMLAM assay showed sensitivity 51.6% (95% CI: 47.3-55.9) but specificity 98.0% (95% CI: 94.9-99.4). The Ultra-sensitive electrochemiluminescence LAM assay demonstrated favorable diagnostic accuracy, potentially offering rapid tuberculosis diagnosis for HIV-negative individuals with pulmonary TB.
Fjords in Chilean Patagonia are highly dynamic systems shaped by land-derived inputs, oceanic exchange, and volcanic activity. Prior to the elaboration of the present article, no in-depth investigation had been undertaken into the anoxic or euxinic conditions of fjords in this region. Consequently, the present research represents an interdisciplinary oceanographic approach to studying Quitralco Fjord (45.6° S, 73.1° W; 2022-2025) and provides the first evidence of a volcanically influenced euxinic fjord in Chilean Patagonia. A subsurface anoxic layer, beginning between 90 and 120 m and extending to the basin floor (~ 160 m), was shown to exhibit elevated temperatures and high concentrations of H2S, consistent with inputs of volcanically derived fluids. A bubble-like acoustic scattering and the detection of CH4 within this layer suggest an external input of the gas into the water column. Although largely stagnant, this layer shifted vertically over time, likely driven by interannual deep-water renewal. Within the euxinic layer, nitrate was completely depleted, while high phosphate (20 μm) and ammonium (25 μm) concentrations indicated an active sulfur cycle. A pronounced deep fluorescence maximum was also detected in the dark, anoxic basin, attributed to fluorescent dissolved organic matter (fDOM) dominated by two humic-like components (C1245(350)-440 and C3270(400)-505) with high aromaticity. Microbial community composition changed markedly across the redox gradient, while geochemical and microbiological fingerprints exhibited shifts in metabolic potential through the water column. Geological emissions of H2S from the seabed and microbial sulfate reduction may contribute to the observed H2S accumulation, enhancing and sustaining euxinic conditions, thereby strongly influencing the basin's biogeochemical cycles. Overall, the present study reveals a previously unrecognized link between volcanic activity and fjord biogeochemistry, documenting for the first time the development of euxinic conditions in a Patagonian fjord in Chile.
Invasive group A streptococcal (iGAS) infections resurged across Europe in late 2022. We aimed to describe the incidence, clinical characteristics, and outcomes of iGAS in Catalonia over two epidemic seasons (2022/23 and 2023/24), and to identify risk factors associated with mortality. A prospective study was conducted including all confirmed iGAS cases reported to the Microbiological Reporting System of Catalonia between October 2022 and October 2024. Epidemiological and clinical data were collected from medical files and incidence rates (IRs) calculated. Multivariable logistic regression was used to assess risk factors associated with mortality. A total of 589 laboratory-confirmed iGAS cases were included; 416 cases (70.6%) from 2022/23 season, and 173 (29.4%) from 2023/24. The overall IR decreased from 5.3 to 2.2 cases per 100,000 population (p < 0.001). In 2022/23, incidence was higher in children (0-14 years) than in adults (9.8 vs. 4.6 per 100,000; p < 0.001), whereas this pattern reversed in 2023/24 (1.7 vs. 2.3; p = 0.028). Pleural empyema predominated in children, while skin and soft tissue infections with bacteraemia in adults. Respiratory viral coinfections were more common in children than in adults (47.1% vs. 10.8%; p < 0.001). Overall, 97.3% of patients were hospitalized, and 26.9% required intensive care (33.3% in children vs. 25.2% in adults; p = 0.08). Overall mortality was 11.3% (12.9% in adults and 5.8% in children: p < 0.05). Multivariable analyses showed age, respiratory viral coinfections, chronic renal disease, and intensive care unit admission independently associated with mortality. The decline in iGAS incidence in 2023/24 was accompanied by a shift in age distribution and a reduction in respiratory viral coinfections, suggesting a return toward pre-pandemic epidemiological patterns. Further research is needed to clarify pathogenic mechanisms and outcome determinants.
The rise in antimicrobial resistance (AMR) is a severe public health threat worldwide. India bears a disproportionately heavy burden of this problem due to ample antimicrobial usage in both humans and animals and scarce integrated surveillance. Since humans, animals, and environmental reservoirs which can harbour resistant microorganisms interact very closely on farms livestock, these are considered critical hotspots for the emergence and dissemination of antimicrobial-resistant bacteria and resistance genes. This work presents a 36-month prospective longitudinal observational study protocol aimed at quantifying the burden and characterizing the transmission dynamics of a selected set of key bacteria, that are clinically significant and hence, pathogenic-Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Streptococcus pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa-alongside their AMRprofiles in livestock, farm-exposed human populations, and environmental reservoirs in Nagpur, India, within the framework of One Health. Seasonal sampling of milk, animal faeces, human stool, soil, wastewater, drinking water, and animal feed will be carried out on dairy farms located in urban, peri-urban, and rural areas. Pathogens will be isolated using standard microbiological techniques and characterized based on antimicrobial susceptibility by employing VITEK®2 and disc diffusion methods. At the same time, bacteriophages against multidrug-resistant isolates will be isolated, purified, and characterized through plaque assays, host-range analysis, electron microscopy, and whole-genome sequencing for their therapeutic potential evaluation. Additionally, metagenomic next-generation sequencing will be utilized on a select number of samples to comprehensively characterize the resistomes and diversity of phages. The research will provide detailed longitudinal data on the frequency and spread of AMR among human, animal, and environmental compartments, create a biobank of AMR isolates and lytic bacteriophages, and offer genomic clues to delineate phage-based treatments and well-informed mitigation strategies of AMR within the framework of One Health in India. The results will be made public through peer-reviewed articles, presentations at scientific meetings, and deposition of sequence data in open-access databases.