Antibacterial activity and mechanism

  • 文章类型: Journal Article
    由于肺炎克雷伯菌(KP)对多种抗生素具有较高的耐药性,它被认为是全球关注的病原体,迫切需要传统抗生素的替代品。已报道3-苯基乳酸(PLA)具有抗食源性细菌的抗微生物活性。然而,没有实验证据表明PLA杀死致病性KP的确切抗菌作用和机制。在这项研究中,牛津杯法表明,PLA对KP有效,最小抑制浓度为2.5mg/mL。此外,PLA以时间和浓度依赖性方式抑制生长和生物膜形成。在体内,PLA能显著提高感染小鼠的存活率,减轻病理组织损伤。进一步探索了PLA对KP的抗菌模式。首先,扫描电子显微镜显示了PLA引起的细胞超微结构的破坏。其次,泄漏的碱性磷酸酶的测量表明PLA破坏了KP的细胞壁完整性,碘化丙啶染色的流式细胞术分析表明PLA破坏了细胞膜的完整性。最后,荧光光谱和琼脂糖凝胶电泳结果表明,PLA与基因组DNA结合并引发其降解。PLA的抗KP作用模式归因于细胞壁的破坏,膜,和基因组DNA结合。这些发现表明,PLA作为抗生素替代品在动物中对抗KP感染的饲料添加剂中具有巨大的潜在应用。
    As Klebsiella pneumoniae (KP) has acquired high levels of resistance to multiple antibiotics, it is considered a worldwide pathogen of concern, and substitutes for traditional antibiotics are urgently needed. 3-Phenyllactic acid (PLA) has been reported to have antimicrobial activity against food-borne bacteria. However, there was no experiment evidence for the exact antibacterial effect and mechanism of PLA kills pathogenic KP. In this study, the Oxford cup method indicated that PLA is effective to KP with a minimum inhibitory concentration of 2.5 mg/mL. Furthermore, PLA inhibited the growth and biofilm formation of in a time- and concentration-dependent manner. In vivo, PLA could significantly increase the survival rate of infected mice and reduce the pathological tissue damage. The antibacterial mode of PLA against KP was further explored. Firstly, scanning electron microscopy illustrated the disruption of cellular ultrastructure caused by PLA. Secondly, measurement of leaked alkaline phosphatase demonstrated that PLA disrupted the cell wall integrity of KP and flow cytometry analysis with propidium iodide staining suggested that PLA damaged the cell membrane integrity. Finally, the results of fluorescence spectroscopy and agarose gel electrophoresis demonstrated that PLA bound to genomic DNA and initiated its degradation. The anti-KP mode of action of PLA was attributed to the destruction of the cell wall, membrane, and genomic DNA binding. These findings suggest that PLA has great potential applications as antibiotic substitutes in feed additives against KP infection in animals.
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  • 文章类型: Journal Article
    目的:本研究旨在确定生姜提取物(GE)的化学成分,评估GE对生姜青枯病病原菌青枯病菌的抗菌活性,并筛选其作用机制。
    结果:使用超高效液相色谱和串联质谱法共鉴定了393个化合物。抗菌试验表明,GE对青枯菌具有较强的抗菌活性,杀菌效果呈剂量依赖性。青枯菌的最低抑菌浓度和最低杀菌浓度分别为3.91和125mg/ml,分别。葛根破坏了青枯菌的细胞膜通透性和完整性,导致细胞内容泄漏,如电解质,核酸,蛋白质,胞外三磷酸腺苷和外多糖。此外,随着GE浓度的增加,青枯菌细胞琥珀酸脱氢酶和碱性磷酸酶的活性逐渐降低。扫描电子显微镜分析显示,GE处理改变了茄尾R.细胞的形态。进一步的实验表明,GE延缓或减缓了生姜上青枯病的发生。
    结论:GE对青枯菌有明显的抗菌作用,抗菌效果是浓度依赖性的。GE治疗改变了形态,破坏膜的渗透性和完整性,降低青枯菌关键酶活性并抑制毒力因子EPS的合成。GE显著控制了生姜侵染过程中的青枯病。
    结论:这项研究深入了解了GE对青枯菌的抗菌机制,这将为GE开辟一个新的应用领域。
    OBJECTIVE: The current study aimed to determine the chemical compositions of ginger extract (GE) and to assess the antibacterial activities of GE against the ginger bacterial wilt pathogen Ralstonia solanacearum and to screen their mechanisms of action.
    RESULTS: A total of 393 compounds were identified by using ultra-performance liquid chromatography and tandem-mass spectrometry. The antibacterial test indicated that GE had strong antibacterial activity against R. solanacearum and that the bactericidal effect exhibited a dose-dependent manner. The minimum inhibitory concentration and minimum bactericidal concentration of R. solanacearum were 3.91 and 125 mg/ml, respectively. The cell membrane permeability and integrity of R. solanacearum were destroyed by GE, resulting in cell content leakage, such as electrolytes, nucleic acids, proteins, extracellular adenosine triphosphate and exopoly saccharides. In addition, the activity of cellular succinate dehydrogenase and alkaline phosphatase of R. solanacearum decreased gradually with an increase in the GE concentration. Scanning electron microscopy analysis revealed that GE treatment changed the morphology of the R. solanacearum cells. Further experiments demonstrated that GE delayed or slowed the occurrence of bacterial wilt on ginger.
    CONCLUSIONS: GE has a significant antibacterial effect on R. solanacearum, and the antibacterial effect is concentration dependent. The GE treatments changed the morphology, destroyed membrane permeability and integrity, reduced key enzyme activity and inhibit the synthesis of the virulence factor EPS of R. solanacearum. GE significantly controlled the bacterial wilt of ginger during infection.
    CONCLUSIONS: This research provides insight into the antimicrobial mechanism of GE against R. solanacearum, which will open a new application field for GE.
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