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Survival of Aging Microbial Populations under Lethal Impacts

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Abstract—

Microorganisms inhabiting natural environments are periodically subject to growth-inhibiting and lethal action of stress factors. Bacterial populations are known to survive and preserve its species identity in the presence of lethal doses of antibiotics due to viable nondividing dormant persister cells. Since the mechanisms of population survival under lethal and especially combined action of chemical and physical stress agents are unknown, this was the goal of the present work. Comparative analysis of survival of the stationary phase cultures of Escherichia coli К-12 MG1655 under lethal impacts of ciprofloxacin (CIP) and a lysing solution, as well as of heat and alkaline shocks, was carried out. Higher resistance to stress impacts was shown for (1) aging stationary phase cells containing a developed biocrystalline nucleoid, compared to the cells just entering the stationary phase; and (2) the populations incubated in a nutrients-free buffer, rather than in a growth medium. CIP treatment of the populations of statically incubated cultures revealed their higher resistance to extreme CIP concentrations (100 µg/mL) compared to the biocidal dose (10 µg/mL) (the Eagle effect). The subpopulations surviving the lethal impacts of heat and pH shock were found to exhibit heterogeneity due to reversion to growth of the cells temporary lacking ability to form colonies on solid media (after 10‒30 days of incubation). Resistance of the cells surviving the heat or pH stresses to the subsequent biocidal treatment with CIP suggests their identification as persister cells. Heterogeneity of the persister subpopulation in their stress resistance may be important for adjustment of the modes for sterilization and antibiotic therapy.

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Funding

This work was supported by the Ministry of Science and Higher Education of the Russian Federation in terms of the State Assignment titled “Microbiology of Innovative Biotechnologies” (no. 122040800164-6) allocated for the Biotechnology Federal Research Center of the Russian Academy of Sciences.

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  1. Winogradsky Institute of Microbiology, Research Centre of Biotechnology, Russian Academy of Sciences, 119071, Moscow, Russia

    G. I. El-Registan, N. G. Loiko & Yu. A. Nikolaev

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  1. G. I. El-Registan
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Correspondence to G. I. El-Registan.

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 Translated by A. Oleskin

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El-Registan, G.I., Loiko, N.G. & Nikolaev, Y.A. Survival of Aging Microbial Populations under Lethal Impacts. Microbiology 91, 693–703 (2022). https://doi.org/10.1134/S0026261722601774

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