Droplet-based methodology for investigating bacterial population dynamics in response to phage exposure.
Droplet-based methodology for investigating bacterial population dynamics in response to phage exposure.
复制标题
基于液滴的研究噬菌体暴露后细菌种群动态的方法。
DOI：
10.3389/fmicb.2023.1260196
复制
发表时间：
2023
影响因子：
5.2
通讯作者：
中科院分区：
文献类型：
--
作者：
全球抗菌素耐药性的惊人上升促使人们努力寻找抗生素治疗的替代品。使用噬菌体，对人类无害的细菌病毒，代表了一种有潜力治疗细菌感染的有前途的方法（噬菌体疗法）。基于显微镜的单细胞技术的最新进展使研究人员能够开发新的定量方法来评估细菌和细菌之间的相互作用，特别是细菌根除细菌病原体种群和调节细菌和病原体种群生长的能力。在这里，我们结合联合收割机液滴微流体与荧光延时显微镜表征细菌大肠杆菌局限在液滴中的生长和裂解动力学时，挑战噬菌体。我们研究了促进感染E.大肠杆菌细胞中分离到一种具有DNA基因组的噬菌体T7（Escherichia virus T7）和两种具有RNA基因组的噬菌体MS 2（Emesvirus zinderi）和Qβ（Qubevirus durum）。我们的微流体捕获装置产生并固定了皮升大小的液滴，从而能够在温度控制的设置中对细菌生长和裂解进行稳定成像。记录细菌种群大小的时间信息长达25小时，使我们能够确定细菌种群的生长速率，并帮助我们揭示噬菌体感染的程度和速度。从长远来看，新型微流体单细胞和群体水平方法的发展将加速研究，以从根本上理解快速噬菌体诱导裂解的遗传和分子基础以及细菌-噬菌体动力学的生态进化方面，并最终帮助确定影响噬菌体治疗成功的关键因素。
An alarming rise in antimicrobial resistance worldwide has spurred efforts into the search for alternatives to antibiotic treatments. The use of bacteriophages, bacterial viruses harmless to humans, represents a promising approach with potential to treat bacterial infections (phage therapy). Recent advances in microscopy-based single-cell techniques have allowed researchers to develop new quantitative methodologies for assessing the interactions between bacteria and phages, especially the ability of phages to eradicate bacterial pathogen populations and to modulate growth of both commensal and pathogen populations. Here we combine droplet microfluidics with fluorescence time-lapse microscopy to characterize the growth and lysis dynamics of the bacterium Escherichia coli confined in droplets when challenged with phage. We investigated phages that promote lysis of infected E. coli cells, specifically, a phage species with DNA genome, T7 (Escherichia virus T7) and two phage species with RNA genomes, MS2 (Emesvirus zinderi) and Qβ (Qubevirus durum). Our microfluidic trapping device generated and immobilized picoliter-sized droplets, enabling stable imaging of bacterial growth and lysis in a temperature-controlled setup. Temporal information on bacterial population size was recorded for up to 25 h, allowing us to determine growth rates of bacterial populations and helping us uncover the extent and speed of phage infection. In the long-term, the development of novel microfluidic single-cell and population-level approaches will expedite research towards fundamental understanding of the genetic and molecular basis of rapid phage-induced lysis and eco-evolutionary aspects of bacteria-phage dynamics, and ultimately help identify key factors influencing the success of phage therapy.
登录
查看更多内容
DOI：
10.3390/ph14111157
发表时间：
2021-11-13
期刊：
Pharmaceuticals (Basel, Switzerland)
影响因子：
--
作者：
Abedon ST;Danis-Wlodarczyk KM;Alves DR
通讯作者：
Alves DR
影响因子：
9.8
作者：
Attrill EL;Claydon R;Łapińska U;Recker M;Meaden S;Brown AT;Westra ER;Harding SV;Pagliara S
通讯作者：
Pagliara S
影响因子：
3.9
作者：
Barizien, A.;Jammalamadaka, M. S. Suryateja;Baroud, Charles N.
通讯作者：
Baroud, Charles N.
影响因子：
16.6
作者：
Chait R;Ruess J;Bergmiller T;Tkačik G;Guet CC
通讯作者：
Guet CC
非表面附着的细菌骨料：无处不在的第三次生活方式。
影响因子：
5.2
作者：
Cai YM
通讯作者：
Cai YM