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Published on: February 23, 2021
Resuscitation dynamics reveal persister partitioning after antibiotic treatment
Xin Fang1,2, Kyle R Allison1,2
1Wallace H. Coulter Department of Biomedical Engineering, Emory University and Georgia Institute of Technology, Atlanta, GA, USA.
Abstract:
Bacteria can survive antibiotics by forming dormant, drug-tolerant persisters. Persisters can resuscitate from dormancy after treatment and prolong infections. Resuscitation is thought to occur stochastically, but its transient, single-cell nature makes it difficult to investigate. We tracked the resuscitation of individual persisters by microscopy after ampicillin treatment and, by characterizing their dynamics, discovered that Escherichia coli and Salmonella enterica persisters resuscitate exponentially rather than stochastically. We demonstrated that the key parameters controlling resuscitation map to the ampicillin concentration during treatment and efflux during resuscitation. Consistently, we observed many persister progeny have structural defects and transcriptional responses indicative of cellular damage, for both β-lactam and quinolone antibiotics. During resuscitation, damaged persisters partition unevenly, generating both healthy daughter cells and defective ones. This persister partitioning phenomenon was observed in S. enterica, Klebsiella pneumoniae, Pseudomonas aeruginosa, and an E. coli urinary tract infection (UTI) isolate. It was also observed in the standard persister assay and after in situ treatment of a clinical UTI sample. This study reveals novel properties of resuscitation and indicates that persister partitioning may be a survival strategy in bacteria that lack genetic resistance.
Insights
Bacteria persisters, dormant cells that prolong infections, resuscitate exponentially, not randomly. This process, influenced by antibiotic concentration and efflux, generates both healthy and damaged daughter cells, a key survival strategy.
Area of Science:
- Microbiology
- Bacterial Physiology
- Antibiotic Resistance
Background:
- Bacteria form dormant, drug-tolerant persisters to survive antibiotic treatment.
- Persisters can reactivate (resuscitate) after therapy, leading to persistent infections.
- The stochastic nature of persister resuscitation has made it challenging to study at the single-cell level.
Purpose of the Study:
- To investigate the dynamics of individual persister resuscitation using microscopy.
- To identify key factors influencing persister resuscitation and progeny viability.
- To explore the phenomenon of persister partitioning across different bacterial species and clinical isolates.
Main Methods:
- Microscopic tracking of individual persister cells after ampicillin treatment in Escherichia coli and Salmonella enterica.
- Characterization of resuscitation dynamics, including ampicillin concentration and efflux.
- Analysis of persister progeny for structural defects and transcriptional responses indicative of cellular damage.
Main Results:
- Persister resuscitation in E. coli and S. enterica follows an exponential, rather than stochastic, pattern.
- Key resuscitation parameters include ampicillin concentration during treatment and efflux during resuscitation.
- Persister progeny often exhibit structural defects and transcriptional damage, with uneven partitioning of these defective cells.
Conclusions:
- Persister resuscitation is an exponential process, not random, with significant implications for infection dynamics.
- Persister partitioning, generating both healthy and damaged progeny, appears to be a bacterial survival strategy, especially in the absence of genetic resistance.
- This phenomenon is conserved across multiple bacterial species and relevant to clinical settings, including urinary tract infections.
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