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首页> 外文期刊>Applied Microbiology >Investigating the Responses of Cronobacter sakazakii to Garlic-Drived Organosulfur Compounds: a Systematic Study of Pathogenic-Bacterium Injury by Use of High-Throughput Whole-Transcriptome Sequencing and Confocal Micro-Raman Spectroscopy
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Investigating the Responses of Cronobacter sakazakii to Garlic-Drived Organosulfur Compounds: a Systematic Study of Pathogenic-Bacterium Injury by Use of High-Throughput Whole-Transcriptome Sequencing and Confocal Micro-Raman Spectroscopy

机译：调查阪崎肠杆菌对大蒜驱动的有机硫化合物的反应：使用高通量全转录组测序和共聚焦显微拉曼光谱法对致病性细菌损伤的系统研究

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We present the results of a study using high-throughput whole-transcriptome sequencing (RNA-seq) and vibrational spectroscopy to characterize and fingerprint pathogenic-bacterium injury under conditions of unfavorable stress. Two garlic-derived organosulfur compounds were found to be highly effective antimicrobial compounds against Cronobacter sakazakii , a leading pathogen associated with invasive infection of infants and causing meningitis, necrotizing entercolitis, and bacteremia. RNA-seq shows changes in gene expression patterns and transcriptomic response, while confocal micro-Raman spectroscopy characterizes macromolecular changes in the bacterial cell resulting from this chemical stress. RNA-seq analyses showed that the bacterial response to ajoene differed from the response to diallyl sulfide. Specifically, ajoene caused downregulation of motility-related genes, while diallyl sulfide treatment caused an increased expression of cell wall synthesis genes. Confocal micro-Raman spectroscopy revealed that the two compounds appear to have the same phase I antimicrobial mechanism of binding to thiol-containing proteins/enzymes in bacterial cells generating a disulfide stretching band but different phase II antimicrobial mechanisms, showing alterations in the secondary structures of proteins in two different ways. Diallyl sulfide primarily altered the α-helix and β-sheet, as reflected in changes in amide I, while ajoene altered the structures containing phenylalanine and tyrosine. Bayesian probability analysis validated the ability of principal component analysis to differentiate treated and control C. sakazakii cells. Scanning electron microscopy confirmed cell injury, showing significant morphological variations in cells following treatments by these two compounds. Findings from this study aid in the development of effective intervention strategies to reduce the risk of C. sakazakii contamination in the food production environment and on food contact surfaces, reducing the risks to susceptible consumers.
机译：我们提出了使用高通量全转录组测序（RNA-seq）和振动光谱来表征和指纹识别不利条件下致病性细菌损伤的研究结果。发现两种大蒜来源的有机硫化合物是针对阪崎肠杆菌的高效抗菌化合物，阪崎肠杆菌是与婴儿的侵入性感染相关的主要病原体，可引起脑膜炎，坏死性小肠结肠炎和菌血症。 RNA-seq显示基因表达模式和转录组反应的变化，而共焦显微拉曼光谱法则表征了这种化学胁迫导致细菌细胞中的大分子变化。 RNA-seq分析表明细菌对a烯的反应与对二烯丙基硫的反应不同。具体而言，阿霍烯引起运动相关基因的下调，而二烯丙基硫醚处理则引起细胞壁合成基因表达的增加。共焦显微拉曼光谱显示，这两种化合物似乎具有相同的I相抗菌机制，可与细菌细胞中含硫醇的蛋白质/酶结合，从而产生二硫键拉伸带，但具有不同的II相抗菌机制，显示了其二级结构的变化。蛋白质有两种不同的方式。如酰胺I的变化所反映的，二烯丙基硫醚主要改变α-螺旋和β-折叠，而a烯改变包含苯丙氨酸和酪氨酸的结构。贝叶斯概率分析验证了主成分分析区分已处理和对照阪崎肠杆菌细胞的能力。扫描电子显微镜证实了细胞损伤，在用这两种化合物处理后，显示出细胞中明显的形态变化。这项研究的结果有助于制定有效的干预策略，以减少食品生产环境中和食品接触表面上阪崎肠杆菌污染的风险，从而降低易感消费者的风险。

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