This study aimed to investigate the global genetic diversity, evolutionary relationships, and antimicrobial resistance (AMR) profiles of Klebsiella pneumoniae by performing a comprehensive pan-genome and phylogenetic analysis across worldwide isolates. A total of 72,057 K. pneumoniae genomes were retrieved from the NCBI database, from which 91high-quality representative genomes each from a unique country were selected based on completeness, metadata availability, and sequence quality. Genomic assemblies were assessed using QUAST, annotated with PROKKA, and analyzed for pan-genomic composition and phylogenetic relatedness using standard bioinformatics pipelines. The pan-genome revealed a large accessory component, reflecting extensive genomic plasticity and adaptability. QUAST analysis indicated significant variability in genome size and contig number, while PROKKA annotation identified diverse coding sequences, tRNA, rRNA, and AMR genes. Phylogenetic clustering demonstrated both geographically localized and globally disseminated lineages, suggesting regional adaptation and intercontinental transmission. This study provides a global perspective on the genomic diversity and evolutionary patterns of K. pneumoniae. The widespread presence of AMR determinants underscores the urgent need for continuous genomic surveillance and integration of metagenomic approaches to improve monitoring, infection control, and therapeutic strategies against multidrug-resistant strains.

This study systematically elucidates the mechanisms of echinocandin resistance in Candidozyma auris (C. auris), providing insights into potential therapeutic strategies. Echinocandin susceptibility of clinical bloodstream C. auris isolates was determined using E-test, and resistance-associated mutations were identified by Sanger sequencing. Biofilm formation under antifungal stress was assessed by crystal violet staining and the 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assays. Changes in cell wall components were analyzed using quantitative fluorescence assays, while expression of cell wall biosynthesis-related genes was evaluated by RT-qPCR. Activation of cell wall integrity (CWI) and calcineurin pathways was examined, and the adjunctive effect of calcineurin inhibitors with anidulafungin (ANI) was assessed using disk diffusion and checkerboard assays. Echinocandin resistance in the clinical isolate C. auris 01 was attributed to an S639F mutation in FKS1. ANI treatment-induced cell aggregation and enhanced biofilm formation. It also triggered cell wall remodeling, increasing chitin, mannan, and β-glucan levels, accompanied by upregulation of β-glucan synthase (FKS1, FKS2), chitin synthase (CHS1, CHS3), and mannan synthesis (PMR1, PMT1). Furthermore, ANI activated both CWI and calcineurin pathways, indicating adaptive stress responses. Notably, calcineurin inhibition exhibited an additive effect with ANI. These results highlight a multifaceted resistance framework involving structural, signaling, and potential genetic alterations.

Polycystic ovary syndrome (PCOS) is a common gynecological and cardiometabolic disorder in reproductive-aged women. Recently gut microbiota alterations have been identified as key contributors to PCOS pathophysiology, but it remains understudied in Indian population. 16S rRNA gene amplicon sequencing using Illumina-MiSeq platform was conducted in 57 PCOS and 30 control women. Diversity indices were assessed and significantly altered taxa were identified by differential abundance tests (Wilcoxon-rank-sum test, ANCOMBC2 and LEfSe) in total and hyperandrogenism-based PCOS subgroups. We quantified levels of short-chain fatty acids (SCFAs) and gut barrier integrity marker, zonulin and LPS by HPLC and ELISA respectively. Our study showed significantly altered beta diversity between PCOS and control groups, driven by hyperandrogenism. PCOS women demonstrate enrichment of Ligilactobacillus species, not previously reported to our knowledge, along with Collinsella species and markedly reduced SCFA-producing taxa, Oscillospiraceae. Additionally, levels of zonulin and fecal butyric acid, a potent SCFA, were significantly altered in PCOS women. Significantly altered taxa correlated with gonadotropin and SHBG levels and were involved in important metabolic pathways in total and PCOS subgroups. Our study offers new insights into the mapping of gut microbiota in PCOS women from western India and implicates hyperandrogenism in driving dysbiosis.

Antibiotic resistance (AMR) poses a significant threat to global health and exerts an increasing economic burden on healthcare systems worldwide. Understanding AMR in human pathogens requires discernment of the complex interactions within the One Health continuum that encompasses humans, animals, and the environment. The issue extends beyond antibiotic misuse in medicine to include a broad spectrum of sector activities that contribute to the development and spread of ressitance genes; veterinary practice, farming, agriculture, aquaculture, and industrial operations. Emerging bacterial resistance within pathogens of public health importance are examined from the One Health perspective, focusing on the mechanisms of genetic change that enable resistance. AMR in specific bacteria particularly important due to the transmissibility of these pathogens amongst individuals, nosocomial environments, and the population leading to the resurgence of diseases that were previously controlled. Advanced genomic typing methods enhance the ability to delineate bacterial strains, improving surveillance and monitoring efforts. Recognizing the interconnectedness of human, animal, and environmental health is vital for safeguarding the effectiveness of antibiotics. Coordinated action across all sectors within the One Health framework is essential to preserve these critical drugs for future generations.

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Questions surrounding the presence or absence of microbiomes in sites of the human body with at most low bacterial biomass in health are still not fully resolved. We begin with the notion of microbiome as "micro-biome," a community of several species, versus microbiota as simply "living things," and return to the pioneer epoch of biome research, which in one sense could be viewed as beginning already around 1800. Applying the biome notion to bacteria in sites of the human body, we find concordance with more recent attempts to test what is and what is not a (core) microbiome in practice. The biome perspective is then applied to a double question that has been addressed, without a consensus answer so far: (1) does the healthy gallbladder typically have a stable microbiota, and (2) do gallbladder microbiota, in either health or (gallstone) disease, qualify as microbiomes in the sense considered here?

This study aimed to investigate how the presence and absence of S-Layer proteins (SLp) affects bacterial sensitivity to Penicillin-G. The phenotypic feature of strains in the presence and absence of SLp was evaluated using disk diffusion and nitrocefin tests. The β-lactam resistance gene (blaZ) was detected by Polymerase chain reaction (PCR). Viable cell counts were performed at 0, 4, 8, and 12 hours in Penicillin-G-supplemented medium for strains with and without SLp. Pearson's correlation coefficient was used to analyze the relationship between exposure time and viability. In the absence of SLp, an increase in the zone diameter of all strains has been observed. In the viability test, Lpb. plantarum DA225had an initial viability of 8.47 log CFU/mL. After 12 hours without SLp, it lost 4.97, finishing with 3.50 log CFU/mL. Pearson's correlation showed a positive correlation of 0.68 between the 4th and 12th hours, and 0.97 between the 8th and 12th hours. Suggesting results SLp deficiency does not completely affect the survival of the bacterium, it causes it to become physiologically weaker, and that the SLp has an indirect but significant effect on cell viability.

This study aimed to isolate and comprehensively characterize Aspergillus terreus from the thorns of Canthium coromandelicum and evaluate the pharmacological potential of its metabolites. A. terreus was isolated and identified through morphological, microscopic and molecular analyses (98.89% sequence identity, GenBank Accession No. PV069722.1). Ethyl acetate extract (CT3C) underwent phytochemical screening, GC - MS analysis, in silico PASS and ADME predictions, molecular docking and in vitro assays for antioxidant, antimicrobial, anti-inflammatory and cytotoxic activities. Morphology revealed aleurioconidia and cinnamon-brown colonies. Phytochemical screening revealed the presence of phenols, flavonoids, terpenoids, alkaloids and cardiac glycosides. GC - MS identified five bioactive compounds, notably 2, 5-Piperazinedione and 3-(Methyloxiran-2-yl) methanol. In silico studies predicted strong drug-likeness, high gastrointestinal absorption, favorable bioavailability and low toxicity. Molecular docking showed multi-target activity, with 2,5-Piperazinedione binding to Keap1 at -7.5 kcal/mol. CT3C extract exhibited significant antioxidant activity (DPPH IC50 = 487.33 µg/mL, ABTS IC50 = 579.37 µg/mL), potent cytotoxicity against MCF-7 cells (IC50 = 32.31 µg/mL), moderate anti-inflammatory effects and broad-spectrum antimicrobial activity. This is the first report of A. terreus from C. coromandelicum thorns (CT3C) -derived metabolites demonstrate significant pharmacological potential, suggesting their value as natural therapeutic agents.

Biofilm eradication remains a significant challenge in clinical practice and contributes substantially to healthcare costs. Medical devices serve as a common substrate for biofilm formation and are implicated in a considerable proportion of invasive candidiasis cases. Rezafungin, a next-generation echinocandin, exhibits broad-spectrum fungicidal activity against Candida spp. and in vitro studies have confirmed its efficacy against Candida biofilms. A literature review up to July 2025 was conducted, including PubMed, Embase, Web of Science, and gray literature, focusing on the real-world use of rezafungin for device-related and other biofilm-associated Candida infections. Real-world data may suggest its clinical effectiveness in managing complex fungal infections frequently associated with biofilms, such as implant-associated infections, endocarditis, or osteoarticular infections, particularly in co-morbid patients. Among the 18 reviewed cases, mostly coming from Europe and the United States, a successful outcome was reported in 83.3% of patients after a median treatment duration of over 3 months, with no serious drug-related adverse events reported. Although these findings should be interpreted cautiously, given the limited sample size and heterogeneity of cases, they support rezafungin as a potential therapeutic option for biofilm-associated fungal infections.

Antimicrobial resistance (AMR) remains one of the most significant threats to global health, largely driven by inappropriate antibiotic use. This review outlines the current principles and practices of antimicrobial stewardship (AMS) in the context of critical care. We address the global burden of AMR, the development of resistance and the implementation of AMS strategies from multidisciplinary, diagnostic and pharmacokinetic perspectives. The use of rapid diagnostics, biomarkers such as procalcitonin, and the integration of electronic health records are discussed as enablers of targeted therapy and improved outcomes. Special populations, such as transplant recipients and critically ill patients with sepsis, are examined in the context of AMS, with attention to therapy optimization and resistance prevention. Challenges in implementation - especially in low-resource settings - are addressed, highlighting the need for global cooperation and infrastructure investment. Through evidence-based strategies, stewardship programs aim to enhance antimicrobial efficacy, minimize resistance development, and improve patient outcomes. This review advocates for the responsible use of antimicrobials across all care settings and emphasizes the necessity of continuous education, surveillance and adaptation in AMS policy and practice.