BACKGROUND:Streptococcus pneumoniae remains a major global health threat, causing diseases ranging from mild respiratory infections to severe conditions like pneumonia, sepsis, and meningitis. Although pneumococcal conjugate vaccines (PCVs), including PCV7, PCV10, and PCV13 have significantly reduced disease burden, especially in children, S. pneumoniae continues to exhibit high serotype and genetic diversity. Whole-genome sequencing (WGS) analysis offers high-resolution insights into clonal lineages and multidrug-resistant strains. However, genomic data on Malaysian isolates remain limited. METHODS:This study characterized the whole genome features and comparative profiles of seven invasive S. pneumoniae isolates from two tertiary hospitals in Malaysia. WGS analyses described serotype, sequence type (ST), antimicrobial resistance determinant genes, pan-genome structure, and recombination events. RESULTS:The average genome size was ∼2.12 Mbp, with 1 988-2 205 coding sequences. WGS-based MLST identified five sequence types (ST236, ST320, ST386, ST671, ST695), with ST236 linked to serotypes 19A and 19F related to PMEN clones Taiwan19F-14 and CC271. Core genome analysis with 35 global reference strains revealed three major clades. Notably, isolates TSP95, SSP45, and SSP46 clustered closely with strains from South Korea, suggesting a long-term persistence of ST320 over a decade. Recombination analysis identified both shared and isolate-specific events, forming distinct phylogenetic clusters. Extensive shared recombination was observed in several isolates, while others displayed isolate-specific events, indicating ongoing genetic diversification. CONCLUSION:These findings underscore the critical role of recombination in shaping pneumococcal population structure, evolution, and adaptation.

Attenuated total reflectance Fourier-transform infrared (ATR-FTIR) spectroscopy offers a reagent-minimal route for serum analysis, but full-spectrum chemometric models can exploit non-specific covariance in highly congested biological spectra. This study evaluated a vibrationally constrained chemometric strategy for quantification of routine serum biochemical analytes using assigned-value external quality assessment materials by restricting regression to analyte-linked spectral regions defined a priori and applying day-specific matrix correction with matched-diluent subtraction. Condensed serum external quality assessment materials were analysed as independent dried droplets on a diamond ATR platform. Baseline-corrected, interpolated absorbance profiles within the locked regions were used to train ridge regression models with training-only standardisation. Calibration performance across a six-analyte panel was high, with training R² values of 0.992 to 0.998. External verification against analytical performance specifications showed 95% acceptability for total protein, 100% acceptability for albumin, lactate, and uric acid, 90% acceptability for urea, and lower acceptability for creatinine at 40%. These findings show that vibrational restriction can reduce model degrees of freedom while retaining strong quantitative performance in congested serum spectra. The study supports vibrationally constrained chemometrics as an interpretable ATR-FTIR quantification strategy and provides an external quality assessment-based verification baseline for subsequent evaluation on commutable human serum.

Introduction: The emergence of antibiotic-resistant bacteria, particularly Gram-positive methicillin-resistant Staphylococcus aureus (MRSA), along with methicillin-susceptible S. aureus (MSSA) and Gram-negative Escherichia coli (E. coli), highlights the critical need for new antibacterial approaches. Herbal medicines, particularly Moringa oleifera leaf extract (MOLE), have demonstrated potential in combating bacterial infections. This study assesses the antibacterial efficacy of MOLE against MRSA, MSSA, and E. coli, with a focus on its effectiveness against both resistant and non-resistant bacterial strains. Methods: The antimicrobial susceptibility of ethanolic extract of MOLE was evaluated using the Kirby-Bauer disk diffusion method at five different concentrations (50-800 mg/mL) against MRSA, MSSA and E. coli. The phytochemical composition of MOLE was analysed using liquid chromatography-mass spectrometry (LC-MS). Results: MOLE demonstrated dose-dependent antibacterial susceptibility against both MSSA and MRSA. For MSSA, inhibition zones increased from 11.40 ± 0.65 mm at 100 mg/mL to 21.67 ± 0.75 mm at 800 mg/mL. MRSA exhibited similar dose-response, with inhibition zones expanding from 9.33 ± 0.65 mm at 100 mg/mL to 17.00 ± 0.65 mm at 800 mg/mL, comparable to the positive control, cefoxitin (18.75 ± 0.65 mm). Notably, MOLE exhibited no inhibitory effect against E. coli. LC-MS analysis identified bioactive compounds, including flavonoids, alkaloids, phenolics, and glucosinolates, known for their antibacterial properties. Conclusion: MOLE exhibited significant antimicrobial susceptibility against MRSA and MSSA, but was ineffective against E. coli. Future research should aim to elucidate the mechanisms of action of MOLE, evaluate its safety and efficacy in vivo, and explore potential synergistic interactions with conventional antibiotics.

Ensuring the authenticity and traceability of food products is increasingly critical to combat food fraud, ensure consumer safety, and meet religious dietary requirements. Traditional nucleic acid (NA)-based methods like PCR provide high sensitivity but are restricted to laboratory environments, highlighting the need for portable alternatives. CRISPR-Cas systems have emerged as promising next-generation tools for food authentication due to their unique trans-cleavage activity, enabling efficient detection of NAs. Recent developments emphasize the integration of CRISPR-Cas with portable platforms such as lateral flow assays, nanopore sensors, smartphone-based devices, and microfluidic systems, which support rapid and reliable on-site detection of food contaminants. Moreover, the combination of CRISPR-Cas with advanced technologies could further enhance the reliability and innovation of food safety diagnostics. This review presents an overview of CRISPR-Cas mechanisms, classification, and advantages for NA detection, while also examining advancements in portable CRISPR-integrated systems for field applications. Additionally, it addresses current challenges, regulatory issues, and future directions, including multiplex detection, microfluidics integration, artificial intelligence applications, and eco-friendly diagnostic development. By consolidating these insights, the review provides valuable guidance for researchers, industry professionals, and policymakers, highlighting the transformative potential of CRISPR-Cas systems in advancing precision food authentication within the global supply chain.

Streptococcus pneumoniae is a significant human pathogen globally, associated with various pneumococcal infections, implicated in conditions such as pneumonia, meningitis, otitis media and sepsis. The pathogen employs numerous virulence factors, such as capsular polysaccaharide (CPS), choline binding protein (CBP), lipoproteins and pneumolysin, to engage with and evade host immune response. Recent studies have emphasised the significance of extracellular membrane vesicles (EMVs) in pathogenesis and immunity, making them potential targets for innovative treatment and vaccination approaches. This review provides a comprehensive overview of EMVs derived from S. pneumoniae and delves into their immunogenic potential as a promising novel vaccine candidate against pneumococcal infections. This review describes the mechanisms of EMVs biogenesis, their various immunogenic properties and their capability to influence host immune responses. This review synthesises findings of the past 10 years (2014-2024) regarding S. pneumoniae EMVs, emphasising their biogenesis, composition, immunogenic properties and roles in host-pathogen interactions. Besides, this review also elaborates on EMV-based vaccines and the challenges associated with their advancement. The research underscores the diverse functions of EMVs derived from S. pneumoniae in pathogenesis, encompassing biofilm formation, immunological evasion and host cell damage. EMVs exhibit strong immunogenic characteristics capable of provoking both innate and adaptive immune responses. Pre-clinical studies have indicated EMVs as a broad-spectrum vaccination candidate, overcoming the constraints of existing serotype-specific vaccines. Nonetheless, challenges such as variations in EMVs’ composition, safety issues and production scalability persist as considerable barriers. EMVs are promising strategies in the global control of pneumococcal infections because of their ability to elicit cross-serotype immunity, yet further study is required to solve safety and manufacturing issues.

Objective: This study provides the first systematic synthesis of the burden of Group B Streptococcus (GBS) colonization and invasive disease in Nigeria, with emphasis on prevalence, serotypes, and sequence types (STs). Method: This systematic review and meta-analysis were conducted in accordance with the PRISMA guidelines and was registered on PROSPERO (CRD420251155310). Searches were conducted across multiple databases, including Scopus, ScienceDirect, Web of Science, PubMed, Dimensions, and African Journals Online, as well as in Google Scholar and Google to identify relevant articles. In total, 426 records were retrieved, of which 43 studies met the inclusion criteria. A random-effects model was applied to estimate the pooled prevalence. Result: The pooled prevalence of GBS colonization in Nigeria was 12.0% (95% CI: 9.0–15.0%). Higher colonization rates were observed in Southern Nigeria (13.0%) than in Northern Nigeria (9.0%). The neonatal colonization rate was 16.0%. Colonization rates were 13.0% in pregnant women and 8.0% in non-pregnant individuals. Human immunodeficiency virus status showed no significant association with GBS colonization among pregnant women (OR = 1.47, p = 0.17). Invasive GBS disease was uncommon (3.0%) and occurred only in neonates. Across included studies, serotypes V and II were the most frequently reported, with ST19, ST182, and ST28 being the predominant STs. Conclusions: GBS colonization is common in Nigeria, with marked regional variation and heightened neonatal vulnerability to invasive GBS infections. Notably, nineteen states lacked surveillance data, highlighting substantial gaps in national monitoring. These findings highlight the importance of strengthening prevention strategies, expanding surveillance coverage, and implementing maternal screening and immunization programs to mitigate the burden of GBS.

The global surge in antibiotic resistance, particularly methicillin-resistant Staphylococcus aureus (MRSA), presents a significant public health challenge and emphasises the necessity for alternative therapeutic strategies. Moringa oleifera is rich in bioactive phytochemicals with established antimicrobial properties, however, the impact of various extraction solvents on its efficacy against MRSA has not been thoroughly investigated. This study offers a direct comparison of ethanolic, methanolic and aqueous M. oleifera leaf extracts (MOLE) to identify the optimal solvent for enhancing antibacterial and antioxidant activities. MOLE was extracted through maceration, and antibacterial activity against MRSA was assessed using colony-forming unit (CFU) assays at concentrations of 50 mg/mL, 100 mg/mL and 200 mg/mL, with vancomycin serving as the positive control. Antioxidant capacity was quantified using the ferric reducing antioxidant power (FRAP) assay, while phytochemical profiles characterised via liquid chromatography-mass spectrometry (LC-MS). The ethanolic extract exhibited the highest bioactivity, achieving complete MRSA inhibition at 100 mg/mL and demonstrating efficacy statistically comparable to vancomycin at 50 mg/mL. It also showed significantly greater antioxidant capacity across all tested concentrations. LC-MS analysis linked this enhanced activity to a richer composition of flavonoids and phenolic acids in the ethanolic extract. In summary, ethanol emerged as the most effective solvent for extracting antibacterial and antioxidant compounds from M. oleifera leaves. The findings underscore the potential of ethanolic MOLE as a natural therapeutic candidate against MRSA, warranting further investigation in preclinical models.

ATR-FTIR spectroscopy offers a reagent-minimal route for serum biochemical analysis, but quantitative modelling in serum is complicated by spectral congestion and the risk that unrestricted chemometric models exploit broad matrix covariance rather than analyte-linked information. This study evaluated a restriction-first chemometric workflow in which analyte-linked spectral windows were defined a priori using bounded vibrational localization and then modelled using ridge regression. Condensed serum chemistry external quality assessment materials with provider-assigned target values were analysed by diamond ATR-FTIR after matched-diluent correction. A 7-level training ladder was constructed from three real anchor vials and four volume-weighted intermediate levels, while independent validation used held-out real EQA vials. The analyte panel comprised total protein, albumin, urea, creatinine, lactate, and uric acid. Training R² values ranged from 0.9917 to 0.9975, while held-out validation R² values ranged from 0.9443 to 0.9803. Replicate-level APS pass rates were 100.0% for albumin, 95.8% for urea and lactate, 91.7% for uric acid, 87.5% for total protein, and 83.3% for creatinine. Coefficient and feature-level analyses indicated chemically readable behaviour within predefined spectral windows, with albumin and urea showing the strongest overall profiles. These findings support vibrationally constrained ridge regression as an interpretable chemometric strategy for ATR-FTIR quantification in spectrally congested serum-like matrices and provide a baseline for later validation in commutable human serum.

Streptococcus agalactiae, or Group B Streptococcus (GBS) sequence type 283 (ST283), has emerged as a foodborne zoonotic pathogen of increasing concern, particularly in Southeast Asia. It gained global recognition following a major outbreak in Singapore in 2015, where it caused severe invasive infections such as meningitis, septic arthritis, and bacteremia, even in previously healthy adults, and was linked to the consumption of raw freshwater fish. Since then, outbreaks and sporadic cases of ST283 have been reported in China and Southeast Asia, with incidental reports in a growing number of countries. While ingestion of contaminated raw freshwater fish remains the primary transmission pathway, recent findings have identified a strong link between infections and handling of freshwater fish, as well as the possibility of contamination due to human shedding and human-to-human transmission. Although ST283 remains largely susceptible to first-line antibiotics, delayed recognition in resource-limited settings hampers timely response. This review synthesizes current knowledge on ST283 as an emerging zoonotic and foodborne threat, summarizing its epidemiology, transmission, pathogenesis, diagnosis, and treatment, while highlighting research gaps and the need for strengthened surveillance and preventive measures.

Streptococcus pneumoniae possess extracellular membrane vesicles (EMVs), which acts as carriers for various bacterial proteins. However, their impact on cellular processes and their interactions with the immune system remain poorly comprehended. This study evaluated EMVs as target proteins and immunomodulators to evoke immune responses in humans. EMV from five different serotypes of S. pneumoniae (6A, 14, 19A, 19F, and 23F) were extracted using the ultracentrifugation, ultrafiltration and iodixanol gradient fractionation methods. The protein components were identified using LC-based proteomic analysis and the protein antigenicity was predicted. A total of 61 proteins were identified; of which, 38 were cytoplasmic origin, three extracellular proteins, three of the cell wall regions, 10 from the membrane, and seven unknown origins. Of these 61 proteins, 12 proteins were virulence factors [aminopeptidase, aminopeptidase (pepC), aminopeptidase (pepN), aminopeptidase PepS (pepS), lipoprotein, oligoendopeptidase F (pepF1_1), PavB protein (pavB), peptidase M13 superfamily (pepO), Pneumococcal surface protein PspA (pspA), pyruvate oxidase (spxB), trypsin-like serine protease (htrA) and zinc metalloprotease ZmpB (ZmpB)]. Six proteins were identified as immunogenic (lipoprotein, pavB, pspA, spxB, htrA, and ZmpB), with a high predictive value for the protective antigen. In conclusion, EMVs could lead to the development of a promising strategy to develop safer and more efficient vaccines to combat pneumococcal infections.

The Indian house crow (Corvus splendens) is a species that has rapidly disseminated across Europe, the Middle East, Eastern Africa, the islands in the Indian Ocean, East Asia, Australia, and the Americas, significantly expanding its ecological range. Corvus splendens is a significant invasive species with a diminished genetic diversity due to founder effects. Its reliance on human waste in urban environments increases its ability to spread germs. The crow is ecologically versatile, thriving in anthropogenic environments and consuming waste, thereby promoting its rapid proliferation and competition with indigenous avian species. It uses urban structures for nesting and communal roosting, demonstrates resistance to noise, and assembles near human settlements for food. The present review synthesised information from multiple articles to offer a full overview of C. splendens, with a particular focus on genetic research, its pathogenicity, ecological roles, and behaviour. The main goal of this study was to provide significant insights and describe research gaps for the efficient management of this invasive species.

Halal certification is vital for confirming the halal status of food products, particularly for Muslim consumers. Instances of halal fraud, including the use of counterfeit halal logos and the incorporation of non-halal ingredients in food products, negatively impact Muslim consumers. Gelatine, widely used in the food industry, varies in acceptability depending on its source, with porcine-derived gelatine strictly prohibited in Islam. Therefore, this research aims to identify the presence of porcine DNA in gelatine-containing food products by targeting mitochondrial cytochrome b (cytb) via a real-time polymerase chain reaction (qPCR) assay and to compare the qPCR findings with the sources of gelatine and the presence of a halal logo. Nine food products without a recognised JAKIM halal logo or bearing a foreign halal logo not recognised by JAKIM were purchased from a local supermarket in Sibu, Sarawak. DNA was extracted from the food products using the DNeasy Mericon Food kit, analysed with a spectrophotometer, and used as template DNA in the qPCR assay. Positive qPCR findings were validated through DNA sequencing and BLAST analysis. Of the nine products tested, eight contained detectable porcine DNA, including one product labelled with a halal logo. Sequencing data confirmed Sus scrofa as the predominant species, with BLAST identities ranging from 86.52% to 100% against the NCBI database. In summary, these results highlight the risks posed by mislabelling and fraudulent halal logos, and emphasise the importance of rigorous certification, transparent labelling, and routine molecular verification to protect Muslim consumers and maintain trust in halal markets.

Streptococcus agalactiae, also called Group B Streptococcus (GBS), is a major cause of several infectious diseases in humans and fish. However, the likelihood of GBS transmission between different host species leading to a potential zoonotic problem is less well studied. Understanding the proteomics of GBS is crucial for elucidating its zoonotic potential and developing effective diagnostic and prevention strategies. This study utilized comparative proteomic analysis using LC–MS/MS technique to investigate the similarities and differences between GBS isolates from human and fish sources. The core proteome was found to be highly conserved, with 82.5

Objective: To assess the burden of Group B Streptococcus (GBS) and analyze the distribution of serotypes in relation to their source. The review highlights data gaps in transmission dynamics and regional food consumption practices, which are essential for designing effective public health strategies and advancing vaccine development. Methods: Searches were conducted in Web of Science, MEDLINE, Science Direct, PubMed, and Scopus databases to find studies related to GBS during 1990-2025. Eligible studies were those that described prevalence, serotype distribution or sequence type (ST) of GBS in Southeast Asian countries. Random-effects meta-analysis was used to pool data. Results: A total of 26 studies met the inclusion criteria from eight countries. The pooled estimate of maternal GBS colonization was 15.1%, with serotypes III, V, II, VI, and I a accounting for the majority of cases (91.24%) in the Southeast Asia studies. Data on ST was limited; however, ST1 was found to be predominant in Malaysia and Thailand, while ST283 was notably linked to the consumption of raw fish. Conclusions: The pooled estimate of the maternal colonization with GBS was 15.1% which is equivalent to many other primary and review reports worldwide. Distribution of serotype and ST is needed to be studied in Southeast Asian countries to devise effective preventive measures. These findings underscore the importance of surveillance and tailored prevention strategies to combat GBS infections in Southeast Asia.

Staphylococcus arlettae is an emerging opportunistic pathogen associated with bovine mastitis, a significant concern in animal health and milk production. This study investigates the genomic characteristics of a multidrug-resistant S. arlettae strain isolated from a healthy cow in Malaysia, providing crucial insights into its potential for pathogenicity and spread. The Staphylococcus arlettae BK2L15 isolate was obtained from a nasal swab of a healthy cow in Kelantan. Initial identification was based on its growth characteristics on Mannitol Salt Agar, followed by antimicrobial susceptibility testing using the disc diffusion method. Subsequent identification with matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF/MS) confirmed the isolate as Staphylococcus arlettae, with a score value of 2.09. Whole genome sequencing was conducted using the Illumina MiSeq system, with raw read quality evaluated through QUAST and genome assembly performed using SPAdes v3.12.0. The Staphylococcus arlettae BK2L15 exhibited a multidrug resistance phenotype towards Erythromycin, Cefoxitin, Fusidic Acid, Oxacillin, Penicillin, Azithromycin and Amoxicillin. Whole genome analysis revealed a genome size of 2,699,512 bp with a GC content of 33.50%, assembled into two contigs. The genome comprises 2,915 protein-coding sequences includes all genomes features. The sequencing data of this strain provide a valuable reference for future fine-scale comparative genomic studies, facilitating the establishment of genomic relationships between lineages and enabling the prediction of virulence factors, mobile genetic elements, and antimicrobial resistance genes.

The resistance of ESBLs-producing Kp to various groups of antibiotics commonly used against infections they caused had become a global threat and required urgent attention. This study assessed the extended spectrum beta-lactamases (ESBLs)-producing Klebsiella pneumoniae isolates in terms of their genomic resistance. An analytical profile index (API) 20E kit was used to confirm a total of 100 clinical isolates of ESBL Klebsiella pneumoniae. The disc diffusion method was used to perform the antimicrobial susceptibility testing (AST), which was followed by the phenotypic detection of ESBLs. Six profiled representative ESBL positive strains were subjected to whole genome sequencing (WGS), multilocus sequence typing (MLST), and phylogenetic tree construction using the sequence data. The study showed that 46(46%) of the 100 isolates were positive for ESBL production and antibiotic susceptibility testing revealed significant resistance to β-lactam antibiotics including monobactam especially ampicillin/sulbactam (40%), cephalosporin groups (cefuroxime, cefotaxime, and ceftriaxone) stood at 51%, 49% and 48% respectively and aztreonam with 49%. The WGS analysis of the representative strains revealed genes encoding resistance to aminoglycoside (StrA4, StrB1, aac(3')-IIa, aac(6')-1b, aac(6')1b-cr-1, aadA16, aph(3')-VIa and aadA15), trimethoprim (dfrA14 and dfrA27), sulphonamide (sul1_11, sul2_2 and sul2_3), quinolone (QnrB40-1, QnrB10, QnrS2, OqxA and OqxB), tetracycline (tet(A)_4), fosfomycin (fosA3, floR2 and fosA7), macrolid (mph(A)_1), rifampicin (ARR-3), β-lactam (blaCTX-M-15_23, blaCTX-M-55, blaSHV-1_22, blaSHV11_18, blaSHV-11, blaSHV-1_1.1, blaSHV-11_3, blaSHV-11_19, blaTEM-1_1, blaTEM-1_5, blaOXA-51_10, blaOXA-30_1, blaNDM-1, blaLEN6, blaLEN8 and blaLEN21 were detected. The MLST analysis revealed two novel sequence types of representative strains (2 with ST NF* and 12 with ST NF) and four other heterogeneous STs which include ST394, ST985, ST17 and ST11 while the phylogenetic tree of the strains showed closed clonal relationship and lineages with other reference isolates. In conclusion, the study's results showed a high prevalence of ESBL-producing Kp in the study area, and the representative strains' genomic contents demonstrated that ESBL-producing Kp in a clinical setting could serve as a reservoir for resistance genes and be the source of genetic transfer to other bacterial species. As a result, ongoing surveillance is required to monitor this endemic situation to prevent an epidemiological outbreak of K. pneumoniae- carrying ESBL.

The increasing prevalence of antimicrobial-resistant (AMR) bacteria in humans, animals, and the environment underscores the necessity for a rapid, sensitive, and specific method to identify resistance genes. Objectives: This study aims to develop a reliable detection tool for identifying the tetracycline-resistant gene tet(M) in Enterococcus species using a real-time loop-mediated isothermal amplification (RT-LAMP) assay. Real-time visualization through a turbidimeter enabled precise estimation of time-to-positivity for gene detection. Methodology: Six primers were designed using PrimerExplorer v.5, and the assay was optimized across different temperatures and incubation times. Validation was conducted by testing 52 tet(M)-positive clinical enterococci isolates and spiking urine samples from a healthy volunteer and a cow with tet(M)-positive Enterococcus species. Results: The tet(M) gene was detected as early as 33 min, with optimal amplification occurring within 60 min at 60 °C. The assay demonstrated 100% specificity with the established primers. The sigmoidal graphs were corroborated with visual confirmation methods, including a green color change (visible to the naked eye), green fluorescence (under UV light), and a 200 bp PCR product observed via agarose gel electrophoresis. Notably, the tet(M) RT-LAMP assay exhibited a detection limit of 0.001 pg/μL, significantly surpassing conventional PCR, which had a detection limit of 0.1 pg/μL. Conclusions: This rapid, cost-effective, highly sensitive, and specific tet(M) RT-LAMP assay holds significant promise as a surveillance tool for antimicrobial resistance monitoring within a One Health framework, particularly in low-resource countries.

The blood–brain barrier (BBB) is an essential protective structure that preserves the homeostasis of the central nervous system (CNS) by controlling the transfer of chemicals and infections from the bloodstream into the tissues of the brain. The BBB is primarily composed of brain microvascular endothelial cells (BMECs), which are closely linked by tight junction (TJ) proteins, including occludin, claudins and junctional adhesion molecules (JAMs). However, certain bacterial infections, such as those caused by Escherichia coli ( E. coli ), Neisseria meningitidis ( N. meningitidis ), Streptococcus pneumoniae ( S. pneumoniae ) and Haemophilus influenzae ( H. influenzae ), have developed strategies to compromise these TJs, facilitating their invasion of the CNS and resulting in meningitis. These bacteria utilise several virulence factors, including outer membrane proteins, pili and toxins, to modify host cell signalling pathways, compromise the BBB and facilitate their translocation across barriers. This study aims to clarify the molecular processes employed by these pathogens to penetrate the BBB, with an emphasis on their effects on TJ integrity. A comprehension of these mechanisms is essential for formulating effective medicines to prevent and cure bacterial meningitis by safeguarding or restoring BBB integrity.

Background:Streptococcus pneumoniae (pneumococcus) is an opportunistic pathogen that causes severe upper and lower respiratory tract infections, leading to life-threatening diseases. This study aims to determine the genetic variation of serotypes among a collection of clinical S. pneumoniae isolates using multilocus sequence typing (MLST) and multilocus variable-number tandem repeat analysis (MLVA). Method:A total of 103 viable isolates were serotyped and subjected to MLVA; only those with discrete serotypes (n = 91) were subjected to MLST analysis. Results:The comparative phylogenetic analysis resulted in the segregation of clonal complexes (CC) and 36 singletons accordingly. The major clonal complex, MLST CC320 (n = 23; 25.3%), had a close association with the Taiwan19F-14 clone, consisting of ST236, ST271, and ST320. The second largest group, MLST CC9 (n = 12; 13.2%), had an association with the England14-9 clone, comprising ST7 and ST9. MLVA analysis demonstrated its ability to differentiate subgroups within CCs that share the same sequence type (ST) with distinct MLVA types (MTs). Conclusion:The integration of MLST and MLVA in this study serves as a model for pneumococcal surveillance indicating some well-known globally circulating clones that have been persisting at this study setting.