Byju Arjun S, Patel Deeti, Chen Weijie, Mani Sridhar
Department of Medicine, Genetics, and Molecular Pharmacology, Albert Einstein College of Medicine Bronx, NY, USA.
Bio Protoc. 2021 May 5;11(9):e4008. doi: 10.21769/BioProtoc.4008.
Swarming - swift movement across a surface via flagella propulsion - is a unique property of many bacteria. The role of swarming, particularly among bacterial populations of the human gut microbiome, is not yet fully understood; although, it is becoming an area of increased scientific and clinical inquiry. To further characterize bacterial swarming in human health, an effective assay for swarming that utilizes complex material, such as fecal matter, is necessary. Until now, the vast majority of swarming assays have only been able to accommodate bacteria grown in culture, most often . These assays tend to use a standard lysogenic broth (LB) agar medium; however, the reagents involved have not been tailored to the inoculation of complex material. In this paper, we offer a specialized protocol for eliciting the swarming of bacteria from frozen human fecal samples. We describe the simple, yet reproducible steps required to perform the assay, identifying an ideal volume of 7.5 μl for inoculation of material, as well as an ideal agar concentration of 0.4%. This protocol typically allows researchers to identify swarming within 24 h after incubation in a standard incubator.
群体运动——通过鞭毛推进在表面快速移动——是许多细菌的一种独特特性。群体运动的作用,尤其是在人类肠道微生物群的细菌群体中,尚未完全了解;尽管如此,它正成为一个科学和临床研究日益增加的领域。为了进一步描述人类健康中的细菌群体运动,需要一种有效的利用复杂物质(如粪便)的群体运动检测方法。到目前为止,绝大多数群体运动检测只能检测在培养基中生长的细菌,最常见的是……。这些检测往往使用标准的溶原肉汤(LB)琼脂培养基;然而,所涉及的试剂并非针对接种复杂物质而定制。在本文中,我们提供了一种从冷冻的人类粪便样本中诱导细菌群体运动的专门方案。我们描述了进行该检测所需的简单但可重复的步骤,确定了接种物质的理想体积为7.5微升,以及理想的琼脂浓度为0.4%。该方案通常使研究人员能够在标准培养箱中孵育24小时后识别出群体运动。