亚MIC链霉素和四环素增强广州金黄色葡萄球菌-SAU749生物膜形成的转录组学深入研究

IF 5.9 Q1 MICROBIOLOGY Biofilm Pub Date : 2023-09-20 DOI:10.1016/j.bioflm.2023.100156
Junyan Liu , Tengyi Huang , Zhenbo Xu , Yuzhu Mao , Thanapop Soteyome , Gongliang Liu , Chunyun Qu , Lei Yuan , Qin Ma , Fang Zhou , Gamini Seneviratne
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引用次数: 0

摘要

金黄色葡萄球菌是一种主要的人类病原体,是一种潜在的“超级细菌”,也是一种典型的生物膜形成细菌。随着大量抗生素的使用,临床环境中残留的抗生素使金黄色葡萄球菌的定植、发病机制和耐药性进一步复杂化。本研究旨在研究不同类型抗生素治疗的临床金黄色葡萄球菌分离株生物膜形成过程中的表型和全局基因表达变化。首先,从大量先前鉴定的金黄色葡萄球菌分离株中筛选出一个分离株Guangzhou-SAU749,该分离株在生物量和活力方面表现出较弱的生物膜形成。其次,使用9种常用的治疗金黄色葡萄球菌感染的抗生素,以及10种浓度范围为1/128-4的最小抑制浓度(MIC)和2倍系列稀释的抗生素,作为不同的抗生素应激条件。然后,通过结晶紫和MTS测定法测试了金黄色葡萄球菌Guangzhou-SAU749在8小时、16小时、24小时和48小时等不同阶段的生物膜形成。第三,在PacBio平台上对广州金黄色葡萄球菌-SAU749进行全基因组测序。第四,由于1/2 MIC四环素(TCY)和1/4 MIC链霉素(STR)处理5小时后生物膜形成增强,因此选择相关生物膜样品并进行RNA-seq和生物信息学分析。最后,通过逆转录定量聚合酶链反应(RT-qPCR)检测两组分系统(TCS)和生物膜相关基因在亚MIC TCY和STR处理的生物膜样品中的表达。尽管大多数抗生素在高于MIC的浓度下降低了广州-SAU749生物膜的生物量和细胞活力,但某些抗生素,包括TCY和STR,在亚MIC下促进了生物膜的形成。此外,在关键时间点对TCY和STR的亚MIC处理的生物膜样品进行基因组测序、RNA-seq和RT-qPCR后,鉴定出与TCS和生物膜形成相关的基因lytR、arlR、hssR、tagA、clfB、atlA和cidA有助于增强生物膜形成,为进一步控制金黄色葡萄球菌生物膜形成提供了理论依据。
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Sub-MIC streptomycin and tetracycline enhanced Staphylococcus aureus Guangzhou-SAU749 biofilm formation, an in-depth study on transcriptomics

Staphylococcus aureus is a major human pathogen, a potential “Super-bug” and a typical biofilm forming bacteria. With usage of large amount of antibiotics, the residual antibiotics in clinical settings further complicate the colonization, pathogenesis and resistance of S. aureus. This study aimed at investigating the phenotypical and global gene expression changes on biofilm formation of a clinical S. aureus isolate treated under different types of antibiotics. Firstly, an isolate Guangzhou-SAU749 was selected from a large sale of previously identified S. aureus isolates, which exhibited weak biofilm formation in terms of biomass and viability. Secondly, 9 commonly prescribed antibiotics for S. aureus infections treatment, together with 10 concentrations ranging from 1/128 to 4 minimum inhibitory concentration (MIC) with 2-fold serial dilution, were used as different antibiotic stress conditions. Then, biofilm formation of S. aureus Guangzhou-SAU749 at different stages including 8 h, 16 h, 24 h, and 48 h, was tested by crystal violet and MTS assays. Thirdly, the whole genome of S. aureus Guangzhou-SAU749 was investigated by genome sequencing on PacBio platform. Fourthly, since enhancement of biofilm formation occurred when treated with 1/2 MIC tetracycline (TCY) and 1/4 MIC streptomycin (STR) since 5 h, the relevant biofilm samples were selected and subjected to RNA-seq and bioinformatics analysis. Last, expression of two component system (TCS) and biofilm associated genes in 4 h, 8 h, 16 h, 24 h, and 48 h sub-MIC TCY and STR treated biofilm samples were performed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Although most antibiotics lowered the biomass and cell viability of Guangzhou-SAU749 biofilm at concentrations higher than MIC, certain antibiotics including TCY and STR promoted biofilm formation at sub-MICs. Additionally, upon genome sequencing, RNA-seq and RT-qPCR on biofilm samples treated with sub-MIC of TCY and STR at key time points, genes lytR, arlR, hssR, tagA, clfB, atlA and cidA related to TCS and biofilm formation were identified to contribute to the enhanced biofilm formation, providing a theoretical basis for further controlling on S. aureus biofilm formation.

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来源期刊
Biofilm
Biofilm MICROBIOLOGY-
CiteScore
7.50
自引率
1.50%
发文量
30
审稿时长
57 days
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