家禽市场中具有 CRISPR-Cas 的多药耐药大肠杆菌中 ZnO 纳米粒子的潜力。
Potential of ZnO nanoparticles for multi-drug resistant Escherichia coli having CRISPR-Cas from poultry market in Lahore.
机构信息
Institute of Microbiology, Faculty of Veterinary Science, University of Veterinary and Animal Sciences, Lahore, Pakistan.
Department of Clinical Studies, Faculty of Veterinary and Animal Sciences, Pir Mehr Ali Shah-Arid Agriculture University, Rawalpindi, Pakistan.
出版信息
BMC Microbiol. 2024 Sep 19;24(1):355. doi: 10.1186/s12866-024-03462-7.
BACKGROUND AND OBJECTIVES
Apart from known factors such as irrational use of antibiotics and horizontal gene transfer, it is now reported that clustered regularly interspaced short palindromic repeats (CRISPR) are also associated with increased antimicrobial resistance. Hence, it is critical to explore alternatives to antibiotics to control economic losses. Therefore, the present study aimed to determine not only the association of CRISPR-Cas system with antibiotic resistance but also the potential of Zinc Oxide nanoparticles (ZnO-NPs) for avian pathogenic Escherichia coli (APEC) isolated from poultry market Lahore.
MATERIALS AND METHODS
Samples (n = 100) were collected from live bird markets of Lahore, and isolates were confirmed as Escherichia coli (E. coli) using the Remel One fast kit, and APEC was identified using PCR. The antibiotic resistance pattern in APEC was determined using the minimum inhibitory concentration (MIC), followed by genotypic confirmation of antibiotic-resistant genes using the PCR. The CRISPR-Cas system was also identified in multidrug-resistant (MDR) isolates, and its association with antibiotics was determined using qRT-PCR. The potential of ZnO-NPs was evaluated for multidrug-resistant (MDR) isolates by MIC.
RESULTS
All isolates of APEC were resistant to nalidixic acid, whereas 95% were resistant to chloramphenicol and 89% were resistant to streptomycin. Nineteen MDR APEC were found in the present study and the CRISPR-Cas system was detected in all of these MDR isolates. In addition, an increased expression of CRISPR-related genes was observed in the standard strain and MDR isolates of APEC. ZnO-NPs inhibited the growth of resistant isolates.
CONCLUSIONS
The findings showed the presence of the CRISPR-Cas system in MDR strains of APEC, along with the potential of ZnO-NPs for a possible solution to proceed. This highlights the importance of regulating antimicrobial resistance in poultry to reduce potential health consequences.
背景与目的
除了不合理使用抗生素和水平基因转移等已知因素外，现在有报道称，成簇规律间隔短回文重复序列（CRISPR）也与抗菌药物耐药性增加有关。因此，探索抗生素替代品来控制经济损失至关重要。因此，本研究旨在不仅确定 CRISPR-Cas 系统与抗生素耐药性的相关性，还确定氧化锌纳米粒子（ZnO-NPs）对禽致病性大肠杆菌（APEC）的潜在作用，该菌分离自拉合尔的家禽市场。
材料与方法
从拉合尔的活禽市场采集样本（n=100），使用 Remel One 快速试剂盒确认分离物为大肠杆菌（E. coli），并使用 PCR 鉴定 APEC。使用最低抑菌浓度（MIC）确定 APEC 的抗生素耐药模式，然后使用 PCR 对耐药基因进行基因型确认。在多重耐药（MDR）分离物中也鉴定了 CRISPR-Cas 系统，并使用 qRT-PCR 确定其与抗生素的相关性。通过 MIC 评估 ZnO-NPs 对多重耐药（MDR）分离物的潜在作用。
结果
所有 APEC 分离物均对萘啶酸耐药，而 95%对氯霉素耐药，89%对链霉素耐药。本研究发现了 19 株 MDR APEC，并且在所有这些 MDR 分离物中都检测到了 CRISPR-Cas 系统。此外，在 APEC 的标准株和 MDR 分离物中观察到 CRISPR 相关基因的表达增加。ZnO-NPs 抑制了耐药分离物的生长。
结论
研究结果表明，CRISPR-Cas 系统存在于 APEC 的 MDR 菌株中，同时 ZnO-NPs 具有潜在的解决方案。这强调了在禽类中规范抗菌药物耐药性的重要性，以减少潜在的健康后果。
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