Exploration of immunomodulatory antileishmanials of plant origin is now being strongly recommended to overcome the immune suppression evident during visceral leishmaniasis (VL) and high cost and toxicity associated with conventional chemotherapeutics. In accordance, we assessed the in vitro and in vivo antileishmanial and immunomodulatory potential of ethanolic fractions of Azadirachta indica leaves (ALE) and seeds (ASE). A. indica fractions were prepared by sequential extraction of the powdered plant parts in hexane, ethanol and water. Erythrosin B staining was employed to appraise the anti-promastigote potential of ALE and ASE. Cytostatic or cytocidal mode of action was ascertained and alterations in parasite morphology were depicted under oil immersion light microscopy. Study of apoptotic correlates was performed to deduce the mechanism of induced cell death and anti-amastigote potential was assessed in Leishmania parasitized RAW 264.7 macrophages. In vivo antileishmanial effectiveness was evaluated in L. donovani infected BALB/c mice, accompanied by investigation of immunomodulatory potential of ALE and ASE. Adverse toxicity of the bioactive fractions against RAW macrophages was studied by MTT assay. In vivo side effects on the liver and kidney functions were also determined. Plant secondary metabolites present in ALE and ASE were analysed by Gas chromatography–mass spectrometry (GC-MS). ALE and ASE (500 μg ml−1) exhibited leishmanicidal activity in a time- and dose-dependent manner (IC50 34 and 77.66 μg ml−1, respectively) with alterations in promastigote morphology and induction of apoptosis. ALE and ASE exerted appreciable anti-amastigote potency (IC50 17.66 and 24.66 μg ml−1, respectively) that was coupled with profound in vivo therapeutic efficacy (87.76% and 85.54% protection in liver and 85.55% and 83.62% in spleen, respectively). ALE exhibited minimal toxicity with selectivity index of 26.10 whereas ASE was observed to be non-toxic. The bioactive fractions revealed no hepato- and nephro-toxicity. ALE and ASE potentiated Th1-biased cell-mediated immunity along with upregulation of INF-γ, TNF-α and IL-2 and decline in IL-4 and IL-10 levels. GC-MS analysis revealed several compounds that may have contributed to the observed antileishmanial effect. Dual antileishmanial and immunostimulatory efficacy exhibited by the bioactive fractions merits their use alone or as adjunct therapy for VL.

Vancomycin-resistant enterococci (VRE) are resistant to most classes of antibiotics. Diagnosis of VRE using standard methods takes 2 to 5 days. Development of a rapid PCR-assay that detects and identifies resistant genes in bacteria would provide time-critical information on the presence of VRE in clinical samples allowing early treatment and management of infected patients. Investigate the use of high resolution melting analysis (HRMA) and 16S-rRNA-PCR approach for rapid and cost-effective identification of VRE. Descriptive antibiotic susceptibility studies. Manchester Academic Health Sciences Centre and School of Translational Medicine, University of Manchester, UK, and Department of Clinical Laboratory Sciences, Taibah University, Saudi Arabia. PCR-HRMA using 16S-rRNA V1-primers was used to detect and identify VRE. DNA from different strains of vancomycin-resistant and -sensitive Enterococcus faecalis (VSE) and Enterococcus faecium were amplified using V1-primer followed by HRMA in a single run. Differentiation of VRE from VSE was based on curve shapes generated against reference organisms (Bacteroides fragilis). Amplification curves and difference plots for VRE and VSE. Difference plots were generated for all vancomycin-resistant and -sensitive E faecalis and E faecium strains by subtracting their fluo.rescence melting profile from that of a reference-species B fragilis. A characteristic curve shape was produced by vancomycin-sensitive E faecalis and E faecium. However, vancomycin-resistant strains of these bacteria were associated with a markedly different curve shape facilitating a clear differentiation. The 16S-PCR-HRMA approach has the potential for detecting vancomycin-resistant E faecium and E faecalis. Data with VRE provide the basis for combining VRE identification with pathogens speciation in a rapid, cheap assay able to identify a pathogen as an Enterococcus and whether it is vancomycin-sensitive or -resistant E faecium or E faecalis in a single PCR and HRMA run. Tested on specific, but not all, reference Enterococcus species and clinical isolates. None.

Bloodstream infections are a challenge to clinicians due to high rates of morbidity and mortality. Moreover, rational antimicrobial therapy, if promptly started, can improve the prognosis of sepsis. The gold-standard blood culture has some limitations and it takes time to complete (about 2-5 days). This leaves the clinicians to start immediately broad-range antimicrobial therapy until microbiology culture’s results available that may lead to complicate situation of patient further. Therefore, there is an urgent need for the development of more advanced diagnostic approaches that can overcome limitations of culture-based methods. This review gives an overview of the BSIs and its epidemiology over several cities and hospitals in Saudi Arabia. It also encompasses the diverse molecular diagnostic modalities developed for the rapid detection and identification of BSIs. Furthermore, the growing understanding of the host’s inflammatory responses to infections has led to the discovery of immunological biomarkers as diagnostic tools for sepsis-related BSI. The rational use of immunological and molecular diagnostic markers in clinical settings may speed up diagnosis and prognosis of septic patients. This review shows that the percentage of BSIs in Saudi Arabia, from 1983 to 2015, ranges from 1% to 11% among patients with confirmed infection. It also indicated that Staphylococcus aureus and coagulase negative staphylococci (CoNS) were the most common Gram-positive pathogens; while Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa were the most Gram-negative ones from patients with BSI. The recommendation of this review is to initiate new surveillance systems of BSI in different regions and hospitals in Saudi Arabia and determining the antibiotic susceptibility profile to design antibiograms that may improve the antibiotic stewardship program for treating BSIs. A comparative study should be conducted for patients suspected to have sepsis, using the standard blood cultures, immunological markers and rapid molecular assays to set up a panel to help diagnose BSIs rapidly. This will allow early and optimal patient management and treatment in Saudi Arabia and therefore decrease overall healthcare costs. Key words: Bloodstream infections, sepsis, molecular diagnosis, inflammatory markers, BSI in Saudi Arabia.

Zoonotic toxocariasis (visceral larva migrans or VLM & ocular larva migrans or OLM) refersto human infection caused by helminths that are not natural human parasites. Toxocariasis is anunderestimated geohelminthic infection may be worldwide.Toxocariasis occurs as a result of human infection with the dog ascarid larvae, Toxocara canisor less commonly, the cat ascarid larvae, T. cati. VLM is mainly a disease of young children,especially those with exposure to playgrounds and sandboxes contaminated by dog or catfeces. While common globally, prevalence in both animals and people is highest in developingcountries. In developed countries, more infections are detected among persons in lower socioeconomicstrata. TVLM & OLM clinical presentations, although most infections are asymptomatic,yet in VLM, which occurs mostly in preschool children, larvae invade multiple tissues(mainly liver, lung, skeletal muscle, or heart) causing nonspecific symptoms as fever, myalgia,weight loss, cough, rashes, hepatosplenomegaly accompanied by hypereosinophilia. Migrationto CNS (neurotoxocariasis) is uncommon and can cause eosinophilic meningoencephalitis andepilepsy. Death can occur with severe cardiac, pulmonary, or neurologic involvement.

Background: Rotavirus gastroenteritis (RVGE) remains a leading cause of severe infant diarrhea worldwide, with growing evidence supporting the role of the gut microbiome in modulating the disease. However, the interplay between early-life demographic factors, the gut microbiome, and their combined impact on RVGE clinical severity remains inadequately characterized, particularly in specific geographic populations. Aim: We aimed to investigate how demographic determinants shape gut microbiome composition and function in RVGE and how these features relate to clinical severity. Methods: In our comprehensive case–control study of 165 infants (120 RVGE cases and 45 healthy controls, aged 0–12 months), we utilized 16S rRNA sequencing combined with advanced statistical modeling and machine learning to investigate how demographic factors influence microbiome composition and clinical outcomes. Results: RVGE cases exhibited significantly reduced bacterial diversity (Kruskal–Wallis, Static = 14.85, p < 0.001) and distinct patterns, with community structure most strongly associated with dehydration severity (PERMANOVA; R² = 0.15, p < 0.001). Substantial taxonomic alterations were identified characterized by depletion of beneficial commensals including Akkermansia (LDA score = 3.8, p < 0.001), Faecalibacterium (Random Forest AUC = 0.82, p < 0.001), and Bifidobacterium (r = −0.42 with breastfeeding, p < 0.001), alongside enrichment of inflammation-associated taxa such as Escherichia-Shigella (WBC; r = 0.49, p < 0.001, and CRP; r = 0.56, p < 0.001), Streptococcus (LDA score = 4.2, p < 0.001), and Staphylococcus. Proteobacteria was the top potential biomarker of severe outcomes (Random Forest AUC = 0.85), with abundance positively correlated with systemic inflammation (CRP: r = 0.51, p = 0.003). Functional predictions revealed increased lipopolysaccharide biosynthesis (ko00540) and reduced butanoate metabolism (ko00650, p < 0.001) in severe disease. Importantly, demographic factors significantly modulated clinical outcomes: cesarean-delivered, formula-fed infants presented the most dysbiotic profiles and experienced 3.2-fold longer hospitalization (95% CI: 1.8–5.6, p < 0.001) than vaginally delivered, breastfed infants did. Conclusions: Collectively, these findings demonstrate that early-life demographic factors potentially shape the gut microbiome composition and function, may influence RVGE severity and recovery trajectories, thus providing candidate biomarkers for risk stratification and identifying targets for microbiota-based interventions.