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
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.
1260196
Nikolic, Nela
88a8f576-d9e2-4eb6-9219-39b7065963d3
Anagnostidis, Vasileios
6901485e-ac71-4cfe-8e84-ea6f7ff94ce0
Tiwari, Anuj
0603ffae-31ae-48a1-9b91-ea0b5ceadcc0
Chait, Remy
ffb64fc4-843f-42f5-8974-8942cd28a4ce
Gielen, Fabrice
c77341af-6e84-468f-a89e-0dcda0a75139
21 November 2023
Nikolic, Nela
88a8f576-d9e2-4eb6-9219-39b7065963d3
Anagnostidis, Vasileios
6901485e-ac71-4cfe-8e84-ea6f7ff94ce0
Tiwari, Anuj
0603ffae-31ae-48a1-9b91-ea0b5ceadcc0
Chait, Remy
ffb64fc4-843f-42f5-8974-8942cd28a4ce
Gielen, Fabrice
c77341af-6e84-468f-a89e-0dcda0a75139
Nikolic, Nela, Anagnostidis, Vasileios, Tiwari, Anuj, Chait, Remy and Gielen, Fabrice
(2023)
Droplet-based methodology for investigating bacterial population dynamics in response to phage exposure.
Frontiers in Microbiology, 14, .
(doi:10.3389/fmicb.2023.1260196).
Abstract
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.
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fmicb-14-1260196
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Published date: 21 November 2023
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The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by the BBSRC grant BB/T011777/1 to FG, by the Wellcome Trust Institutional Strategic Support Funding (WT105618MA) Research Restart Award and Pump-Priming Initiative to NN, by the Royal Society grant RGS/R2/192377 to RC, and by the BBSRC-funded South West Biosciences Doctoral Training Partnership (training grant reference 2578821).
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Local EPrints ID: 487935
URI: http://eprints.soton.ac.uk/id/eprint/487935
ISSN: 1664-302X
PURE UUID: e439541e-dfae-484c-82d3-a48f9d83038a
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Date deposited: 11 Mar 2024 17:36
Last modified: 18 Mar 2024 04:18
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Author:
Nela Nikolic
Author:
Vasileios Anagnostidis
Author:
Anuj Tiwari
Author:
Remy Chait
Author:
Fabrice Gielen
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