Department of Chemistry and Biochemistry, Old Dominion University, Norfolk, VA, USA.
Department of Microbiology and Immunology, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC, USA.
Anaerobe. 2019 Oct;59:205-211. doi: 10.1016/j.anaerobe.2019.102080. Epub 2019 Aug 3.
Regulation of bacterial motility to maximize nutrient acquisition or minimize exposure to harmful substances plays an important role in microbial proliferation and host colonization. The technical difficulties of performing high-resolution live microscopy on anaerobes have hindered mechanistic studies of motility in Clostridioides (formerly Clostridium) difficile. Here, we present a widely applicable protocol for live cell imaging of anaerobic bacteria that has allowed us to characterize C. difficile swimming at the single-cell level. This accessible method for anaerobic live cell microscopy enables inquiry into previously inaccessible aspects of C. difficile physiology and behavior. We present the first report that vegetative C. difficile are capable of regulated motility in the presence of different nutrients. We demonstrate that the epidemic C. difficile strain R20291 exhibits regulated motility in the presence of multiple nutrient sources by modulating its swimming velocity. This is a powerful illustration of the ability of single-cell studies to explain population-wide phenomena such as dispersal through the environment.
调节细菌的运动性以最大程度地获取营养或最小化暴露于有害物质的风险，在微生物的增殖和宿主定殖中起着重要作用。由于对厌氧菌进行高分辨率活细胞显微镜检测的技术困难，阻碍了对艰难梭菌（以前称为梭状芽孢杆菌）运动性的机制研究。在这里，我们提出了一种广泛适用于厌氧细菌活细胞成像的方案，使我们能够在单细胞水平上对艰难梭菌的游动进行特征描述。这种易于实现的厌氧活细胞显微镜方法使我们能够探究之前无法触及的艰难梭菌生理学和行为学方面。我们首次报道了在不同营养物质存在的情况下，营养型的艰难梭菌具有可调节的运动性。我们证明，流行的艰难梭菌 R20291 株通过调节其游动速度，在存在多种营养源的情况下表现出可调节的运动性。这有力地说明了单细胞研究能够解释群体现象（例如通过环境进行分散）的能力。
