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Microfluidic droplet reactors: reveal bacterial community interactions in antibiotics

Zhao, X.; Cuniberti, G.; Bachmann, M.; Ruelens, P.; Farr, A.; de Visser, J. A. G. M.; Baraban, L.

Abstract

Antibiotics are effective in treating infections caused by bacteria and are therefore widely used in clinical practice.1
However, the misuse and abuse of antibiotics have resulted in the emergence of large numbers of drug-resistant microorganisms, leading to a global crisis in the healthcare sector.2, 3
The spread of resistance in bacterial communities is not limited to gene transfer; cross-protection also allows different bacteria in the same environment to coexist through mutual protection in the presence of antibiotics.4
Here, a microfluidic droplet reactor system is used to investigate the bacterial community interaction in an antibiotic environment, tracking the survival status of co-cultured antibiotic-sensitive and strong antibiotic-resistant strains in an antibiotic (Cefotaxime, CTX) environment in various harshness.
The microfluidic reactor system monitors the growth status of two bacterial strains in real-time and high throughput by detecting their different emission fluorescent signals; E.coli YFP (antibiotic-sensitive) produces yellow fluorescent protein and E.coli BFP (strong antibiotic-resistant strain) produces the blue fluorescent protein.5
As the fluorescent intensity change during the incubation of both strains, the growth status of both bacterial strains is recorded. A phenomenon of cross-protection is observed in the low concentration of CTX (0.05-5 µg/mL). In addition, to confirm the effect of cross-protection, cell status is examined using microscopy, as well as studies fluorescence from resuspended cells and β-lactamase activity with a plate reader.
1. Hutchings, M. I., Truman, A. W. and Wilkinson, B., Antibiotics: past, present and future. Curr Opin Microbiol 51, 72-80 (2019).
2. Bell, M., Antibiotic misuse: a global crisis. JAMA Intern Med 174, 1920-1 (2014).
3. Murray, C. J. L., Ikuta, K. S., Sharara, F., et al., Global burden of bacterial antimicrobial resistance in 2019: a systematic analysis. The Lancet 399, 629-655 (2022)
4. Yurtsev, E. A., Conwill, A. and Gore, J. Oscillatory dynamics in a bacterial cross-protection mutualism. PNAS 113, 6236-6241 (2016)
5. Zhao, X., Illing, R., Ruelens, P., et al., Coexistence of fluorescent Escherichia coli strains in millifluidic droplet reactors, Lab on a Chip 11, 1492-1502 (2021)

  • Vortrag (Konferenzbeitrag)
    Seventh International Conference on Multifunctional, Hybrid and Nanomaterials, 17.-22.10.2022, Genoa, Italy

Permalink: https://www.hzdr.de/publications/Publ-36428