Objectives This study investigated the prevalence of AmpC beta‐lactamase‐producing Escherichia coli and Klebsiella pneumoniae in clinical specimens collected in Lahore, Pakistan, and aimed to identify potential inhibitors targeting AmpC beta‐lactamases. Methods A total of 17,500 clinical specimens were collected from June 2018 to June 2021 and analyzed for E. coli and K. pneumoniae . These isolates underwent phenotypic identification using microscopy and the API 20E kit. AmpC beta‐lactamase production was initially screened using cefoxitin disk diffusion and inhibitor‐based methods, while the presence of plasmid‐borne AmpC genes (CIT, EBC, and DHA) was confirmed by PCR. Additionally, the study used computational drug discovery techniques to screen for oxadiazole, quinazolinone, and sulfonylurea derivatives as potential inhibitors of the AmpC protein. Results Our data showed high antibiotic resistance rates among E. coli and K. pneumoniae isolates: 74.5% were resistant to cefotaxime and ceftazidime, and 45% to cefoxitin. PCR testing revealed that the CIT gene was the most prevalent in both E. coli (65.4%) and K. pneumoniae (63%), while other genes (EBC and DHA) and combinations were also present ( p < 0.005). Using computational drug discovery, several compound libraries were screened against the AmpC protein. Tolbutamide showed the most promising results, with strong binding affinity and favorable drug‐like properties. Conclusion Plasmid‐mediated AmpC beta‐lactamase‐producing E. coli and K. pneumoniae were highly prevalent, with the CIT genotype predominating among clinical isolates. Computational screening identified Tolbutamide as a promising lead compound, warranting further experimental validation before therapeutic application.
Background Healthcare workers are a first contact for patients with infectious diseases and may be at occupational risk during hemorrhagic fever outbreaks. The Marburg virus disease (MVD) outbreak in southern Ethiopia, ongoing during data collection in December 2025, highlighted the need to assess healthcare workers’ knowledge, attitudes, and practices regarding MVD. Therefore, this study assessed knowledge, attitudes, and practices and examined factors associated with practice among healthcare work. Methods A cross‐sectional study of 238 healthcare workers at Dilla University Teaching Hospital was conducted from December 1 to December 22, 2025. Descriptive statistics, frequencies, and percentages summarized participant characteristics and KAP responses. Binary logistic regression examined factors associated with good practice. Variables with p value ≤ 0.25 in bivariable analysis were included in the multivariable model. Adjusted odds ratios (AORs) with 95% confidence intervals (CIs) were calculated, and significance was set at p < 0.05. Results About 51.7%, 57.1%, and 55.0% of participants had good knowledge, positive attitudes, and good practice toward MVD, respectively, based on study‐specific cutoffs. Most identified direct contact with blood and body fluids as an MVD transmission route (92.0%). However, gaps were observed regarding environmental risk factors and MVD distribution and origin. Most recognized reporting suspected cases (91.2%), avoiding contact with infected individuals or their possessions (91.2%), and using personal protective equipment (87.8%). In multivariable analysis, modification of working habits from concern about acquiring MVD (AOR = 2.19, 95% CI: 1.21–3.99) and good MVD knowledge (AOR = 1.96, 95% CI: 1.13–3.39) were positively associated with good practice. Conclusion Findings indicate gaps in MVD‐related knowledge and infection prevention among healthcare workers at Dilla University Teaching Hospital. Targeted competency‐based training, simulation, IPC resources, and facility‐level preparedness systems are warranted to strengthen readiness for MVD and other viral hemorrhagic fever events.
Klebsiella pneumoniae (Kpn) is an opportunistic bacterium associated with various diseases, with significant morbidity and mortality due to treatment challenges posed by hypervirulent and multidrug-resistant (MDR) strains. This study investigates the prevalence of antimicrobial resistance, virulence factors, and clonal diversity of Kpn isolates. A total of 704 records were collected from the patient registries of two tertiary-care hospitals in Chattogram, Bangladesh. From these records, 123 biochemically confirmed clinical Klebsiella species were selected from admitted patients for molecular analysis. Antibiotic susceptibility, hypermucoid phenotype, and the presence of AMR genes (sul1, aadB) and the hypervirulence gene (rmpA) were determined. Whole-genome sequencing was also performed on one of the most drug-resistant isolates. Out of all isolates (n = 704), 72.59% were MDR, 18.89% DDR, and 7.95% DSDR. Among MDR, hospital-acquired infections (76%) were more prevalent than community-acquired infections (24%), with higher rates among geriatric (27%) and pediatric (25%) patients. The isolates showed high resistance to third-generation cephalosporins and the highest sensitivity to colistin. PCR analysis of the 81 phoE-positive isolates revealed that 85.18% carried the sul1 gene, while 6.17% harbored the aadB gene. Additionally, 12.34% were positive for the virulence gene rmpA, and 1.24% contained all three genes, sul1, aadB, and rmpA. Among the isolates, 65.43% were string test-positive. Notably, these string test-positive strains were resistant to most tested antibiotics compared to string test-negative strains. Whole-genome sequencing confirmed that KP-CU-1 as K. pneumoniae, with a 5.26 Mbp draft genome and an N50 of 164,530 bp, classified as sequence type 29 (ST29). This study indicates that MDR Kpn is highly prevalent, with a high proportion of hypervirulent strains. Establishing a nationwide AMR surveillance network will be essential to control the spread of MDR and hypervirulent Kpn in Bangladesh.
BackgroundNecrotizing fasciitis (NF) caused by Vibrio species is a rapidly progressing, life-threatening infection. The purposes of this study were to characterize polymicrobial Vibrio NF and compare its clinical features, microbiology, antimicrobial therapy, and outcomes with those of monomicrobial Vibrio NF, and to evaluate the effective usage of empirical antibiotics.MethodsOne hundred and ninety-four patients surgically confirmed with Vibrio NF were retrospectively reviewed, in which 180 (92.8%) patients were infected by Vibrio vulnificus. There were 179 patients (92.3%) with a monomicrobial infection and 15 (7.7%) with polymicrobial infections. Clinical characteristics, outcomes, laboratory data, and antibiotic usage were analyzed and compared between the two groups.ResultsTwenty-nine patients with Vibrio NF died (14.9%). Mortality was 15.6% (28/179) in the monomicrobial group and 6.7% (1/15) in the polymicrobial group. Among the 15 polymicrobial cases, 14 were caused by Vibrio vulnificus and 1 by Vibrio parahaemolyticus. Gout was more frequently observed in the polymicrobial group (4/15) than in the monomicrobial group (11/179). Vibrio isolates showed high susceptibility to third-generation cephalosporins, fluoroquinolones, and tetracyclines. Effective antimicrobial therapy was associated with lower mortality (p = 0.002) and lower amputation rates (p = 0.004).ConclusionsPolymicrobial Vibrio NF was uncommon but was not clinically less severe than monomicrobial Vibrio NF in this 20-year cohort. Patients with polymicrobial infection more frequently had gout, suggesting a potential clinical association that warrants further investigation. These findings indicate that suspected polymicrobial Vibrio NF should be treated with the same urgency as monomicrobial disease, with prompt surgery, intensive care, and empirical antimicrobial therapy covering both Vibrio species and relevant copathogens.
BackgroundThe recent COVID-19 pandemic has resulted in a wide spectrum of neurological complications, including stroke, and emerging evidence suggests a critical role of microRNAs in their development. This study aimed to evaluate the expression levels of miR-124, miR-636, and miR-485-5p in COVID-19 patients with and without stroke, compared with healthy individuals, to investigate their potential roles in COVID-19-associated neurological manifestations.MethodsThis cross-sectional study included 100 individuals: 25 COVID-19 patients with stroke, 25 COVID-19 patients without stroke, and 50 healthy controls. COVID-19 infection was confirmed using RT-PCR. Following plasma isolation and RNA extraction, quantitative real-time PCR was performed to determine the expression levels of miR-124, miR-636, and miR-485-5p. The Kruskal-Wallis test followed by Dunn's post hoc test was used for statistical analysis.ResultsThe miR-124 expression was significantly increased in the stroke-positive group (5.13 +/- 2.48) compared to both the stroke-negative (1.78 +/- 1.03) and control groups (1.69 +/- 0.64) (p = 0.0084). Significant differences were observed between the stroke-positive and stroke-negative groups (p = 0.0156), as well as between the stroke-positive and control groups (p = 0.0333). Although not statistically significant, a trend toward increased levels was observed in the stroke-positive group (p = 0.3594). miR-636 showed no significant differences among the three groups (p = 0.9987). miR-485-5p showed a nonsignificant trend toward higher levels in stroke-positive patients (2.15 +/- 1.53 vs. 1.38 +/- 1.01, p = 0.36).ConclusionThe results demonstrated a significant overexpression of miR-124 in COVID-19 patients with stroke, highlighting its potential role in COVID-19-associated neurological disorders. Further studies are required to elucidate the underlying mechanisms and assess its potential as a diagnostic and therapeutic biomarker.
