The CRISPR System and MepA Multidrug Efflux Pump Linked to Antibiotic Resistance in Staphylococcus aureus.

IF 1.9 2区 农林科学 Q3 FOOD SCIENCE & TECHNOLOGY
Jingge Wang, Panpan Liu, Na Li, Xiaohui Chen, Xiaoqiang He, Guiqin Wang
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Abstract

Staphylococcus aureus (S. aureus) is a major zoonotic pathogen. To investigate CRISPR carriage in S. aureus isolates from cows with mastitis and the role of the CRISPR system and efflux pumps in antibiotic resistance. We analyzed antibiotic resistance genes and CRISPR loci, sequenced spacers, and assessed correlations between CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) presence and antibiotic resistance in 234 S. aureus isolates. The changes in CRISPR sequences were examined by continuous passage of 360 generations without antibiotic pressure. Subsequently, variations in CRISPR loci and transcript levels were measured under ciprofloxacin (CIP) exposure. In addition, an S. aureus-25-mepA was constructed to evaluate changes in antimicrobial sensitivity and mepA transcript levels in both planktonic and biofilm states. Our results revealed a CRISPR loci detection rate of 7.69% among the 234 S. aureus isolates, with significantly lower rates of the antibiotic resistance genes gyrA, grlA, norA, and tet(M) in CRISPR-positive isolates compared to those in CRISPR-negative isolates (p < 0.05). CIP-resistant strains exhibited loss of repeat and spacer sequence in CRISPR loci, and the transcript abundance of these loci gradually decreased under CIP pressures, indicating that CRISPR loci deletion or transcript level downregulation under antibiotic stress may be a potential regulatory mechanism of antibiotic resistance. Correlation analysis linked CIP resistance in both planktonic and biofilm S. aureus to mepA transcript levels and biofilm integrity. Our study provides insight into the mechanism by which S. aureus develops antibiotic resistance via the CRISPR system and the MepA efflux pump, offering a theoretical foundation for monitoring the prevalence and resistance of pathogenic bacteria.

CRISPR系统和MepA多药外排泵与金黄色葡萄球菌抗生素耐药性相关
金黄色葡萄球菌(金黄色葡萄球菌)是一种主要的人畜共患病原体。目的研究奶牛乳腺炎金黄色葡萄球菌分离株的CRISPR携带情况,以及CRISPR系统和外排泵在抗生素耐药性中的作用。我们分析了抗生素耐药基因和CRISPR位点,对间隔序列进行了测序,并评估了234株金黄色葡萄球菌中CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)的存在与抗生素耐药之间的相关性。在没有抗生素压力的情况下,通过连续传代360代来检测CRISPR序列的变化。随后,在环丙沙星(CIP)暴露下测量CRISPR位点和转录物水平的变化。此外,我们构建了金黄色葡萄球菌-25-mepA,以评估浮游和生物膜状态下抗菌敏感性和mepA转录水平的变化。结果显示,234株金黄色葡萄球菌的CRISPR位点检出率为7.69%,其中,CRISPR阳性菌株的耐药基因gyrA、grlA、norA和tet(M)检出率显著低于阴性菌株(p < 0.05)。CIP耐药菌株表现出CRISPR位点重复序列和间隔序列缺失,且这些位点的转录物丰度在CIP压力下逐渐降低,表明抗生素胁迫下CRISPR位点缺失或转录物水平下调可能是抗生素耐药的潜在调控机制。相关分析将浮游和生物膜金黄色葡萄球菌的CIP抗性与mepA转录水平和生物膜完整性联系起来。我们的研究揭示了金黄色葡萄球菌通过CRISPR系统和MepA外流泵产生耐药性的机制,为监测病原菌的流行和耐药性提供了理论基础。
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来源期刊
Foodborne pathogens and disease
Foodborne pathogens and disease 医学-食品科技
CiteScore
5.30
自引率
3.60%
发文量
80
审稿时长
1 months
期刊介绍: Foodborne Pathogens and Disease is one of the most inclusive scientific publications on the many disciplines that contribute to food safety. Spanning an array of issues from "farm-to-fork," the Journal bridges the gap between science and policy to reduce the burden of foodborne illness worldwide. Foodborne Pathogens and Disease coverage includes: Agroterrorism Safety of organically grown and genetically modified foods Emerging pathogens Emergence of drug resistance Methods and technology for rapid and accurate detection Strategies to destroy or control foodborne pathogens Novel strategies for the prevention and control of plant and animal diseases that impact food safety Biosecurity issues and the implications of new regulatory guidelines Impact of changing lifestyles and consumer demands on food safety.
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