Green synthesis of silver nanoparticles derived from lemon and pomegranate peel extracts to combat multidrug-resistant bacterial isolates.

IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal, genetic engineering & biotechnology Pub Date : 2023-10-10 DOI:10.1186/s43141-023-00547-0
Mohamad Abdelrazik, Hassan H Elkotaby, Abdullah Yousef, Ahmed F El-Sayed, Mohamed Khedr
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Abstract

Background: Multidrug-resistant (MDR) bacteria are acknowledged as one of the main factors contributing to chronic illnesses and fatalities globally. Numerous diseases, including bloodstream infections, pneumonia, urinary tract infections, and surgical site infections, can be brought on by MDR bacteria. Therefore, a crucial topic of continuing research is the development of a novel and different treatment for MDR microbial pathogens. This work is introduce an alternative method for elimination of MDR bacterial isolates which are causative agents of urinary tract infection among people in Egypt. In our study, we need a novel strategy to combat MDR bacteria by green-synthesized metal nanoparticles (MNPs). That is due to the ability of MNPs to penetrate the cell wall and the cell membrane of gram-positive and gram-negative bacteria.

Methods: Clinical isolates of MDR bacteria had their antibiotic susceptibility assessed before being molecularly identified using 16 s rRNA, sequencing, and phylogenetic analysis. Also, genetic profiles of isolated strains were performed using ISSR and SDS-PAGE. Finally, characterized plant-mediated silver nanoparticles derived from lemon and pomegranate peel extracts were evaluated against isolated multidrug-resistant bacterial stains.

Results: In our present trial, one-hundred urine samples were collected from 71 females and 29 males complaining of UTI (urinary tract infection) symptoms. One-hundred microbial isolates were isolated, including 88-g negative and only 8-g positive bacteria in addition to four yeast isolates (Candida species). A total of 72% of the isolated bacteria showed MDR activity. The most prevalent MDR bacterial isolates (Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterococcus faecalis, and Klebsiella pneumoniae) were identified through 16S rDNA PCR sequencing as with accession numbers OP741103, OP741104, OP741105, OP741106, and OP741107, respectively. Lemon and pomegranate-mediated silver nanoparticles [Ag-NPs] were characterized by UV spectroscopy, FTIR, XRD, and TEM with average size 32 and 28 nm, respectively. Lemon and pomegranate-mediated silver nanoparticles [Ag-NPs] showed an inhibitory effect on the selected five MDR isolates at MIC 50 and 30 µg/mL, respectively. These common bacterial isolates were also genetically examined using ISSR PCR, and their total protein level was evaluated using SDS-PAGE, showing the presence of distinct genetic and protein bands for each bacterial species and emphasizing their general and protein composition as a crucial and essential tool in understanding and overcoming MDR behavior in UTI patients.

Conclusions: Lemon and pomegranate-mediated silver nanoparticles [Ag-NPs] were found to have an inhibitory effect on MDR isolates. Therefore, the study suggests that [Ag-NPs] could be a potential treatment for MDR UTI infections caused by the identified bacterial species.

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从柠檬和石榴皮提取物中提取的银纳米颗粒的绿色合成,以对抗多重耐药性细菌分离株。
背景:多药耐药(MDR)细菌是全球公认的导致慢性病和死亡的主要因素之一。耐多药细菌可引起许多疾病,包括血液感染、肺炎、尿路感染和手术部位感染。因此,继续研究的一个关键课题是开发一种新的、不同的MDR微生物病原体治疗方法。这项工作介绍了一种消除耐多药细菌分离株的替代方法,耐多药菌株是埃及人尿路感染的病原体。在我们的研究中,我们需要一种通过绿色合成金属纳米颗粒(MNPs)对抗MDR细菌的新策略。这是由于MNPs能够穿透革兰氏阳性菌和革兰氏阴性菌的细胞壁和细胞膜。方法:在使用16s rRNA、测序和系统发育分析进行分子鉴定之前,对耐多药细菌的临床分离株进行抗生素敏感性评估。利用ISSR和SDS-PAGE对分离菌株进行了遗传图谱分析。最后,从柠檬和石榴皮提取物中提取的具有特征的植物介导的银纳米颗粒对分离的耐多药细菌污渍进行了评估。结果:在我们目前的试验中,从71名女性和29名男性中收集了100份尿液样本,他们抱怨UTI(尿路感染)症状。分离出100个微生物分离株,包括88g阴性细菌和仅8g阳性细菌,此外还有4个酵母分离株(念珠菌属)。共有72%的分离细菌显示出MDR活性。通过16S rDNA PCR测序鉴定了最流行的耐多药菌株(大肠杆菌、铜绿假单胞菌、鲍曼不动杆菌、粪肠球菌和肺炎克雷伯菌),登录号分别为OP741103、OP741104、OP741105、OP741106和OP741107。通过紫外光谱、FTIR、XRD和TEM对柠檬和石榴介导的银纳米颗粒[Ag NPs]进行了表征,平均尺寸分别为32和28nm。柠檬和石榴介导的银纳米颗粒[Ag NPs]分别在MIC 50和30µg/mL下对选定的五种MDR分离株显示出抑制作用。这些常见的细菌分离株也使用ISSR PCR进行了基因检测,并使用SDS-PAGE评估了它们的总蛋白质水平,显示了每种细菌都存在不同的遗传和蛋白质带,并强调它们的一般和蛋白质组成是理解和克服尿路感染患者耐多药行为的重要工具。结论:柠檬和石榴介导的银纳米粒子[Ag NPs]对MDR分离株具有抑制作用。因此,该研究表明,[Ag NPs]可能是由已鉴定的细菌引起的MDR UTI感染的潜在治疗方法。
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