Introduction: In Pakistan, one of the main causes of mortality and morbidity is typhoid fever. Notably, extended drug-resistant (XDR) typhoid has been misdiagnosed widely due to many physicians continued use of Widal and Typhi-dot as serological tests for diagnosis during the COVID-19 pandemic. There were increased cases of extensively drug-resistant typhoid due to the use of extended drugs during the COVID-19 pandemic without confirming the antimicrobial susceptibility of Salmonella typhi. Methodology: In this study, 118 patient blood samples after the COVID-19 pandemic was analyzed by automated blood culture through BacT/ALERT 3D, and antimicrobials susceptibility was performed. And compared with the antimicrobial susceptibility before the COVID-19 pandemic samples of the same patients. Results: Out of 118 blood cultures of Post-COVID-19 isolates, 34 cultures were positive for Salmonella typhi. On antibiotic susceptibility test profiling of these 34 isolates, 44.12% were extensive drug-resistant (XDR), 38.24% were multidrug-resistant (MDR), 8.82% were non-MDR, non-XDR and 8.82% were resistant to all antibiotics. While in Pre-COVID-19 isolates 21.74% were XDR, 60.87% were MDR, 17.39% were non-MDR, non-XDR and 0.0% were resistant to all antibiotics. There were an increased XDR typhoid ratio as compared to MDR typhoid in Post-COVID-19 isolates. Conclusions: There is an increased emergence of XDR typhoid cases in Post-COVID-19 isolates of Salmonella typhi as compared to the Pre-COVID-19 Salmonella typhi isolates. Moreover, implementing antimicrobial susceptibility profiling and proper diagnosis before using any antibiotic against Salmonella typhi prevent the XDR typhoid outbreak in the near future.

The extended-spectrum β-lactamase (ESBL) genes have become more common, increasing the prevalence of multidrug-resistant gram-negative bacteria (GNBs) found in clinical patients. This cross-sectional study aimed to find the antimicrobial susceptibility of clinically isolated GNBs and their correlations with ESBL genes. A total of 140 samples were collected without any age discrimination and proceeded for their growth on culture media, gram staining, and biochemical characterization. Antimicrobial susceptibility testing was done by the Kirby-Baur disk diffusion method and PCR was employed for ESBLs associated genes (bla_CTX-M, bla_SHV, bla_OXA, bla_TEM). Out of 140 samples, the majority (60%) were GNBs, with K. Pneumoniae being the most prevalent (38.1%), followed by E. coli (29.8%), P. aeruginosa (15.5%), Proteus spp (10.7%), and Citrobacter spp (6%). These GNBs were resistant to various antibiotics, including cefixime (70.2%), amoxicillin (50%), cefoperazone (21.4%), imipenem (17.9%), ceftaroline (89%), gentamicin (72.6%), tobramycin (64.3%), amikacin (52.4%). Individual ESBL gene frequencies were bla_CTX-M (40.5%), bla_TEM (27%), bla_OXA (21.6%), and bla_SHV (10.8%). Species-wise, ESBLs‐producing genes bla_CTX-M, and bla_TEM were most frequent in K. Pneumoniae (13.5%) and E. coli (27%) while bla_OXA in P. aeruginosa (13.5%) and bla_SHV in Proteus spp (2.7%). In conclusion, bla_CTX-M was the main gene associated with ESBL production in resistance clinical isolates followed by bla_TEM, bla_OXA, and bla_SHV genes. To avoid resistance isolates, hospitals must implement infection control and antibiotic stewardship plans.

Introduction: Vaccines are very Important to prevent disease outbreaks. Vaccines have special importance in the prevention of diseases such as Avian influenza (AI) and Fowl cholera (FC) which are economically significant afflicting poultry in Pakistan as well as around the world. The immunogenicity of an experimental bivalent inactivated Avian Influenza H5 and Fowl Cholera (P. multocida)(PM) vaccine is tested in the field in this study. Methodology: A total of 100-day-old broiler chicks were selected. The chicks were divided into four groups of 25 birds each. Group 1 injected with monovalent H5, Group 2 with PM. Group 3 with Bivalent H5 and P. multocida vaccine in equal dose of both vaccines at the rate of 1:1. Group 4 was kept as control. Group 1, 2 and 3 received 1st Vaccine shot at 7th day of age and 2nd dose at day 14th day of age. Results: The immunization potency by ELISA and protective efficacy by challenge study of a bivalent inactivated fowl cholera and AI vaccine is determined through experimental application on broiler flocks in this study. ELISA test was used to determine the peak of the humeral immune response against PM at the 1st, 3rd, 4th, and 6th weeks after vaccination, same testing for Influenza H5 titer was done by using HI after vaccination. ELISA titer of Fowl cholera in monovalent vaccine group at 6th week was 2900 while in bivalent vaccine group was 4056. Similarly, HI test of H5 titer was 7log in bivalent while in monovalent was 5.3 log in 6th week. Conclusions: It was observed that bivalent vaccine has relatively higher titer (protection) against H5 and PM than monovalent, with same dose (0.5ml dose/ pullet).

Antibiotic resistance among Gram-negative bacteria, particularly those capable of forming biofilms, has become a serious clinical challenge with limited treatment options. In this study, magnesium oxide nanoparticles (MgO NPs) were synthesized using a green, cost-effective approach based on aqueous leaf extract of Cymbopogon citratus (lemongrass) and evaluated for their antibacterial and antibiofilm potential against multidrug resistant (MDR) clinical isolates of Acinetobacter baumannii , Klebsiella pneumoniae , Pseudomonas aeruginosa , and Escherichia coli . Minimum inhibitory concentration (MIC) values determined by broth microdilution ranged from 250 to 500 µg/ml, and minimum bactericidal concentration (MBC) values ranged from 1000 to 2000 µg/ml. Antibacterial activity assessed by the well diffusion method was dose-dependent, with inhibition zones ranging from 18 to 28 mm. The effect of MgO NPs on biofilm formation and eradication of pre-formed biofilms was evaluated using microtiter plate assay with crystal violet staining. Percentage inhibition of biofilm formation ranged from 27 to 97.1%, while eradication of established biofilms ranged from 51.9 to 96.4%. Exposure to sub-inhibitory concentrations of MgO NPs led to a 2 to 3-fold increase in reactive oxygen species (ROS) production in treated cells compared to controls. MgO NPs also significantly reduced the protein, carbohydrate, and DNA content of the biofilm matrix. Swimming, swarming, and twitching motility were markedly reduced in treated isolates. Congo red agar assay confirmed that slime production was inhibited following NP treatment. These findings suggest that biogenically synthesized MgO NPs hold considerable promise as antibacterial and antibiofilm agents against MDR Gram-negative pathogens.

Introduction: Essential oils exhibit potent antimicrobial properties, offering natural and effective solutions for combating a wide range of pathogens. Objective: In this study, Lemongrass (Cymbopogon citratus) Nano emulsion was prepared using Tween80 aqueous solution as the surfactant and various core materials, including pure canola oil and a mixture of lemongrass oil (LG) and canola oil (CA) at different ratios. Materials and Methods: Bacterial isolation from cow milk samples was performed using Mannitol Salt Agar (MSA) followed by incubation for 24 hours. The GC-MS analysis was conducted to determine the chemical composition of the sample. Nano emulsions were characterized by particle size and turbidity. Results: The GC-MS analysis revealed citral (geranial) as the dominant compound (32.4%), followed by neral (31.41%). Nano emulsion particle sizes decreased with increasing concentration, ranging from 31.99 nm (1% w/v) to 40.3 nm (2.5% w/v). Turbidity analysis showed an absorbance value of 0.128 ± 0.05 at 600 nm. In cow milk, the Nano emulsion demonstrated a dose-dependent reduction in microbial count (from 10⁸ CFU/mL in control to 10² CFU/mL at 400 ppm/2 mL). The minimum inhibitory concentration (MIC) of 400 ppm against Staphylococcus aureus confirmed its antimicrobial efficacy. Time-kill dynamics further supported dose-dependent bacterial inhibition. Conclusion: These findings highlight lemongrass essential oil's significant antimicrobial properties, suggesting its potential applications in food preservation and pharmaceutical industries.

Background: Atherosclerosis remains a leading cause of cardiovascular morbidity and mortality worldwide, contributing to life-threatening complications such as coronary artery disease, myocardial infarction, stroke, and peripheral vascular disease. Despite well-established risk factors such as hyperlipidemia, hypertension, oxidative stress, and inflammation, the molecular underpinnings of plaque development and progression are still being elucidated. Among the emerging regulators, microRNAs (miRNAs)—short, non-coding RNAs—have gained recognition for their ability to post-transcriptionally modulate gene expression and influence vascular cell behavior and immune responses. Objective: This review aims to explore the regulatory role of miRNAs in the initiation, progression, and phenotypic diversification of atherosclerotic plaques, and to evaluate their potential as diagnostic biomarkers and therapeutic targets. Main Discussion Points: The review discusses the involvement of miRNAs in endothelial dysfunction, lipid accumulation, foam cell formation, and plaque instability. It highlights the differential expression of key miRNAs such as miR-92a, miR-126, miR-155, and miR-221 across various stages and phenotypes of atherosclerosis. Methodological approaches including next-generation sequencing, in vivo animal models, and integrative omics analyses are reviewed. The role of genetic polymorphisms and environmental stimuli in modulating miRNA expression is also addressed. Conclusion: miRNAs serve as central regulators in the complex pathophysiology of atherosclerosis, with distinct expression profiles correlating with disease stage and vascular phenotype. Their diagnostic and therapeutic potential warrants further investigation through well-designed clinical studies to enable their translation into clinical practice.