期刊
ACS INFECTIOUS DISEASES
卷 8, 期 1, 页码 242-254出版社
AMER CHEMICAL SOC
DOI: 10.1021/acsinfecdis.1c00564
关键词
Escherichia coli; doxycycline; antibiotic resistance; microfluidics; experimental evolution; metagenomic deep sequencing
资金
- Defense Threat Reduction Agency [HDTRA1-15-1-0069]
- National In-stitute of Allergy and Infectious Diseases [R01A1080714]
The use of monodisperse microdroplets in experimental evolution has shown to provide better control over environmental conditions and reveal potential new evolutionary trajectories and dynamics, including large-scale chromosomal rearrangements and amplifications. Microdroplet emulsions, with their advantages in automation and condition control, offer a high-throughput method for biomarker identification and preclinical evaluation.
In vitro experimental evolution of pathogens to antibiotics is commonly used for the identification of clinical biomarkers associated with antibiotic resistance. Microdroplet emulsions allow exquisite control of spatial structure, species complexity, and selection microenvironments for such studies. We investigated the use of monodisperse microdroplets in experimental evolution. Using Escherichia coli adaptation to doxycycline, we examined how changes in environmental conditions such as droplet size, starting lambda value, selection strength, and incubation method affected evolutionary outcomes. We also examined the extent to which emulsions could reveal potentially new evolutionary trajectories and dynamics associated with antimicrobial resistance. Interestingly, we identified both expected and unexpected evolutionary trajectories including large-scale chromosomal rearrangements and amplification that were not observed in suspension culture methods. As microdroplet emulsions are well-suited for automation and provide exceptional control of conditions, they can provide a high-throughput approach for biomarker identification as well as preclinical evaluation of lead compounds.
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