Abstract
Persister cells are growth-arrested subpopulations that can survive possible fatal environments and revert to wild types after stress removal. Clinically, persistent pathogens play a key role in antibiotic therapy failure, as well as chronic, recurrent, and antibiotic-resilient infections. In general, molecular and physiological research on persister cells formation and compounds against persister cells are much desired. In this study, we firstly demonstrated that the spore forming Gram-positive model organism Bacillus subtilis can be used to generate persister cells during exposure to antimicrobial compounds. Interestingly, instead of exhibiting a unified antibiotic tolerance profile, different number of persister cells and spores were quantified in various stress conditions. qPCR results also indicated that differential stress responses are related to persister formation in various environmental conditions. We propose, for the first time to the best of our knowledge, an effective method to isolate B. subtilis persister cells from a population using fluorescence-activated cell sorting (FACS), which makes analyzing persister populations feasible. Finally, we show that alpha-helical cationic antimicrobial peptides SAAP-148 and TC-19, derived from human cathelicidin LL-37 and human thrombocidin-1, respectively, have high efficiency against both B. subtilis vegetative cells and persisters, causing membrane permeability and fluidity alteration. In addition, we confirm that in contrast to persister cells, dormant B. subtilis spores are not susceptible to the antimicrobial peptides.
Highlights
Persisters are phenotypic variations of microbial cells that are tolerant to harsh environments such as antibiotic exposure and starvation
Unlike resistant bacteria that rely on mutations, persisters are non-genetic and non-heritable cellular states that can be triggered by certain stresses or generated spontaneously as a bet-hedge strategy [1]
Persisters can promote the formation of antibiotic resistant bacteria through, for example, their survival advantage [6] or increase the spread of antibiotic resistance plasmids [7]
Summary
Persisters are phenotypic variations of microbial cells that are tolerant to harsh environments such as antibiotic exposure and starvation. Unlike resistant bacteria that rely on mutations, persisters are non-genetic and non-heritable cellular states that can be triggered by certain stresses or generated spontaneously as a bet-hedge strategy [1]. It was believed that all bacterial species can form persisters [4,5]. The existence of persisters leads to incomplete bacterial eradication both in vitro and in vitro, causing recurrent contamination and infection. Persisters can promote the formation of antibiotic resistant bacteria through, for example, their survival advantage [6] or increase the spread of antibiotic resistance plasmids [7].
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