Evaluating The Efficacy of Green-Method-Prepared Zinc Oxide Nanoparticles On Gene Expression of Efflux Pumps in Klebsiella Pneumoniae Bacteria Isolated from Women with Gestational Diabetes

Vaginitis is one of the most common women's health problems, and its severity increases in pregnant women with gestational diabetes due to higher susceptibility to infection and increased rates of antibiotic resistance. The research problem: The increasing prevalence of multidrug-resistant isolates of Klebsiella pneumoniae and its association with AcrAB-TolC ejaculation pumps limits treatment efficacy, while the effect of green-method nanoparticles on gene expression of these pumps remains unclear. This paper aimed to detect the acrA and acrB genes in K. pneumoniae isolates and evaluate the effect of zinc oxide nanoparticles prepared with garlic and rosemary extracts on their gene expression. Two hundred and three vaginal swabs were collected from pregnant women (aged 18–45) 72 hours after antibiotic discontinuation. The samples were cultured, and the isolates were identified using biochemical and Vitek assays. Susceptibility testing was then performed using the disk diffusion method according to CLSI 2025. Conventional PCR was used to detect acrA/acrB, and RT-qPCR was used to calculate expression folds using the 2^−ΔΔCt method with 16S as a reference gene. The results showed bacterial growth in 152 samples, and 44 isolates were from women with gestational diabetes, with K. pneumoniae being 15.9% of the isolates exhibiting MDR-like affinity with high resistance to some cephalosporins and piperacillin (up to 85.7%), and high sensitivity (up to 100%) to some fluoroquinolones. AcrA and acrB were present in all tested isolates. ZnO-NPs reduced acrA expression to 0.60 (garlic) and 0.43 (rosemary), while acrB remained close to unchanged with garlic (1.01) and increased with rosemary to 1.53, indicating a selective effect on ejaculation pump components and their clinical applications