Phan Joann, Ranjbar Saba, Kagawa Miki, Gargus Matthew, Hochbaum Allon Israel, Whiteson Katrine L
Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, CA, United States.
Department of Chemical and Biomolecular Engineering, University of California, Irvine, Irvine, CA, United States.
Front Cell Infect Microbiol. 2020 Oct 6;10:569685. doi: 10.3389/fcimb.2020.569685. eCollection 2020.
infection models are important for studying the effects of antimicrobials on microbial growth and metabolism. However, many models lack important biological components that resemble the polymicrobial nature of chronic wounds or infections. In this study, we developed a perfused meat model that supports the growth of the human pathogen in a native meat microbial background to investigate the impact of antibiotics and hydrogen peroxide on polymicrobial community growth and metabolism. plays an important role as an etiological agent involved in chronic infections and is a common opportunistic pathogen. Chemical stressors in the form of hydrogen peroxide, carbenicillin, and gentamicin were perfused through the meat with polymicrobial growth on the surface. The relative abundances of and the background microbial community were analyzed by cell viability assays, and metabolic changes of the entire community in response to different antimicrobial treatments were characterized by GC-MS analysis of volatile organic compounds. The meat background community was characterized by amplicon sequencing. Relative densities of and background microbiota were similar under control conditions. Antimicrobial stressors, even at sub-inhibitory, physiologically relevant concentrations, spurred dominance of the meat surface community. Volatile metabolite ion intensity levels showed that antibacterial treatments drive changes in microbial metabolism. The abundance of the derived metabolite, acetophenone, remained stable with treatment, whereas the relative abundances of 2-butanone, 2-nonanone, and 2-aminoacetophenone changed in response to treatment, suggesting these could serve as biomarkers of infection. Our model recapitulates some of the physiological conditions of chronic wounds and facilitates high throughput experiments without the high cost of models. Expanded use of this perfusion model will contribute to the understanding of polymicrobial growth and metabolism in the context of chronic wounds and infections.
感染模型对于研究抗菌药物对微生物生长和代谢的影响非常重要。然而,许多模型缺乏重要的生物学成分,这些成分无法模拟慢性伤口或感染的多微生物性质。在本研究中,我们开发了一种灌注肉模型,该模型在天然肉类微生物背景下支持人类病原体的生长,以研究抗生素和过氧化氢对多微生物群落生长和代谢的影响。作为参与慢性感染的病原体,起着重要作用,并且是一种常见的机会性病原体。以过氧化氢、羧苄青霉素和庆大霉素形式存在的化学应激源通过表面有微生物生长的肉类进行灌注。通过细胞活力测定分析目标病原体和背景微生物群落的相对丰度,并通过对挥发性有机化合物的气相色谱 - 质谱分析来表征整个群落对不同抗菌处理的代谢变化。通过扩增子测序对肉类背景群落进行表征。在对照条件下,目标病原体和背景微生物群的相对密度相似。抗菌应激源,即使在亚抑制、生理相关浓度下,也会促使目标病原体在肉类表面群落中占主导地位。挥发性代谢物离子强度水平表明抗菌处理会驱动微生物代谢的变化。衍生代谢物苯乙酮的丰度在处理过程中保持稳定,而2 - 丁酮、2 - 壬酮和2 - 氨基苯乙酮的相对丰度因处理而发生变化,表明这些物质可作为感染的生物标志物。我们的模型概括了慢性伤口的一些生理条件,并有助于进行高通量实验,而无需承担动物模型的高昂成本。这种灌注模型的广泛应用将有助于在慢性伤口和感染的背景下理解多微生物的生长和代谢。