antibioticsАнтибиотики и ХимиотерапияAntibiot Khimioter = Antibiotics and Chemotherapy0235-2990ООО «Издательство ОКИ»10.37489/0235-2990-2020-65-1-2-27-32antibiotics-706Research ArticleВ ПОМОЩЬ ПРАКТИКУЮЩЕМУ ВРАЧУGUIDELINES FOR PRACTITIONERSФормирование персистеров у клинических изолятов K.pneumoniae, индуцированных меропенемом, амикацином и их комбинациейFormation of Persisters in Clinical Isolates of K.pneumoniae Induced with Meropenem, Amikacin, and Their CombinationМаркеловаН. Н.MarkelovaN. N.

Москва

Moscow

sedykhnatali@yandex.ruТутельянА. В.TutelyanA. V.

Москва

Moscow

СедыхН. Г.SedykhN. G.

Москва

Moscow

sedykhnatali@yandex.ruФБУН «Центральный научно-исследовательский институт эпидемиологии» Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека; ФГБУ «Российский научный центр рентгенорадиологии» МЗ РФРоссияScience Central Institute of Epidemiology of the Federal Service on Customers' Rights Protection and Human Well-being Surveillance; Central Research Institute of Roentgenology and Radiology of Ministry of Health of Russian FederationRussian FederationФБУН «Центральный научно-исследовательский институт эпидемиологии» Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека; Первый Московский государственный медицинский университет им. И. М. Сеченова МЗ РФРоссияScience Central Institute of Epidemiology of the Federal Service on Customers' Rights Protection and Human Well-being Surveillance; I. M. Sechenov First Moscow State Medical University of Ministry of Health of Russian FederationRussian FederationФБУН «Центральный научно-исследовательский институт эпидемиологии» Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человекаРоссияScience Central Institute of Epidemiology of the Federal Service on Customers' Rights Protection and Human Well-being SurveillanceRussian Federation202025052020651-22732Copyright © Маркелова Н.Н., Тутельян А.В., Седых Н.Г., 20202020Маркелова Н.Н., Тутельян А.В., Седых Н.Г.Markelova N.N., Tutelyan A.V., Sedykh N.G.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.antibiotics-chemotherapy.ru/jour/article/view/706

Исследование индуцированной антибиотиколерантности чувствительных к меропенему и амикацину клинических изолятов Klebsiella pneumoniae выявило формирование у них фенотипически гетерогенных персистеров, представленных SCV (small colony variant) и NCV (normal colony variant) формами. Корреляции образования персистеров между данными антимикробными препаратами не обнаружено (R=-0,37), что указывает на различные механизмы толерантности к антибиотикам. Способность амикацина индуцировать образование SCV-персистеров была выше, чем у меропенема (p<0,05), и снижению их количества способствовал синергетический эффект обоих препаратов, максимально - до 80,0%. При этом SCV-персистеры K.pneumoniae характеризовались кратковременным повышением адаптивной устойчивости к антибиотикам, которые были использованы в качестве индукторов образования персистирующих фракций бактериий (p<0,05), что может способствовать их более эффективному выживанию в средах с антибиотиками.

A study of induced antibiotic tolerance of clinical isolates Klebsiella pneumoniae sensitive to meropenem and amikacin revealed the formation of phenotypically heterogeneous persisters in them, represented by the SCV (small colony variant) and NCV (normal colony variant) forms. There was no correlation of persister formation between these antimicrobial drugs (R=-0.37), which indicates different mechanisms of tolerance to antibiotics. The ability of amikacin to induce the formation of SCV persisters was higher than that of meropenem (p-level > 0.05), and the synergistic effect of both drugs contributed to a decrease in their number, to a maximum of 80.0%. At the same time, SCV-persists of K.pneumoniae were characterized by a short-term increase in adaptive resistance to antibiotics, which were used as inducers of the formation of persistent bacterium fractions (p-level > 0.05), which may contribute to their more effective survival in media with antibiotics.

антибиотикотолерантностьперсистерыфенотипическая гетерогенностьSCV (вариант малой колонии)antibiotic tolerancepersistersphenotypic heterogeneitySCV (small colony variant)ReferencesFisher R.A., Gollan B., Helaine S. Persistent bacterial infections and per-sister cells. Nature Reviews Microbiology 2017; 15: 453–464.Fisher R.A., Gollan B., Helaine S. Persistent bacterial infections and per-sister cells. Nature Reviews Microbiology 2017; 15: 453–464.Shan Y., Lazinski D., Rowe S., Camilli A., Lewis K. Genetic basis of per-sister tolerance to aminoglycosides in Escherichia coli. mBio 2015; 6 Is 2: e00078-15.Shan Y., Lazinski D., Rowe S., Camilli A., Lewis K. Genetic basis of per-sister tolerance to aminoglycosides in Escherichia coli. mBio 2015; 6 Is 2: e00078-15.Chua S. L., Yam J. K., Hao P., Adav S. S., Salido M. M., Liu Y. et al. Selective labelling and eradication of antibiotic-tolerant bacterial populations in Pseudomonas aeruginosa biofilms. Nature communications 2016; 7: 10750.Chua S. L., Yam J. K., Hao P., Adav S. S., Salido M. M., Liu Y. et al. Selective labelling and eradication of antibiotic-tolerant bacterial populations in Pseudomonas aeruginosa biofilms. Nature communications 2016; 7: 10750.Torrey H. L., Keren I., Via L. E., Lee J. S., Lewis K. High persister mutants in Mycobacterium tuberculosis. PLoSOne 2016; 11 (5): e0155127.Torrey H. L., Keren I., Via L. E., Lee J. S., Lewis K. High persister mutants in Mycobacterium tuberculosis. PLoSOne 2016; 11 (5): e0155127.Haaber J., Friberg C., McCreary M., Lin R., Cohen S. N., Ingmer H. Reversible antibiotic tolerance induced in Staphylococcus aureus by concurrent drug exposure. MBio 2015; 6 Is 1: e02268-14.Haaber J., Friberg C., McCreary M., Lin R., Cohen S. N., Ingmer H. Reversible antibiotic tolerance induced in Staphylococcus aureus by concurrent drug exposure. MBio 2015; 6 Is 1: e02268-14.Keren I., Kaldalu N., Spoering A., Wang Y., Lewis K. Persister cells and tolerance to antimicrobials. FEMS Microbiology Letters 2004; 230 Is 1: 13–21.Keren I., Kaldalu N., Spoering A., Wang Y., Lewis K. Persister cells and tolerance to antimicrobials. FEMS Microbiology Letters 2004; 230 Is 1: 13–21.Wiuff C., Zappala R. M., Regoes R. R., Garner K. N., Baquero F., Levin B. R. Phenotypic tolerance: Antibiotic enrichment of non-inherited resistance in bacterial populations. Antimicrob Agents Chemother 2005; 49: 1483–1494.Wiuff C., Zappala R. M., Regoes R. R., Garner K. N., Baquero F., Levin B. R. Phenotypic tolerance: Antibiotic enrichment of non-inherited resistance in bacterial populations. Antimicrob Agents Chemother 2005; 49: 1483–1494.Fleur D. L., Kumamoto C. A., Lewis K. Candida albicans biofilms produce antifungal-tolerant persister cells. Antimicrob Agents Chemother 2006; 50 (Pt11): 3839–3846.Fleur D. L., Kumamoto C. A., Lewis K. Candida albicans biofilms produce antifungal-tolerant persister cells. Antimicrob Agents Chemother 2006; 50 (Pt11): 3839–3846.Roberts M. E., Stewart P. S. Modelling protection from antimicrobial agents in biofilms through the formation of persister cells. Microbiology 2005; 151: 75–80.Roberts M. E., Stewart P. S. Modelling protection from antimicrobial agents in biofilms through the formation of persister cells. Microbiology 2005; 151: 75–80.Johns B. E., Purdy K. J., Tucker N. P., Maddocks S. E. Phenotypic and genotypic characteristics of small colony variants and their role in chronic infection. Microbiol Insights 2015; 8: 15–23.Johns B. E., Purdy K. J., Tucker N. P., Maddocks S. E. Phenotypic and genotypic characteristics of small colony variants and their role in chronic infection. Microbiol Insights 2015; 8: 15–23.Leimer N. Rachmühl C., Marques M., Bahlmann A. S. et al. Nonstable Staphylococcus aureus small-colony variants are induced by low pH and sensitized to antimicrobial therapy by phagolysosomal alkalinization. J Infect Dis 2015; 213 (2): 305-308.Leimer N. Rachmühl C., Marques M., Bahlmann A. S. et al. Nonstable Staphylococcus aureus small-colony variants are induced by low pH and sensitized to antimicrobial therapy by phagolysosomal alkalinization. J Infect Dis 2015; 213 (2): 305-308.Proctor R. A., Eiff C., Kahl B. C., Becker K., McNamara P., Herrmann M. at al. Small colony variants: a pathogenic form of bacteria that facilitates persistent and recurrent infections. Nat Rev Microbiol 2006; 4 (Pt4): 295–305.Proctor R. A., Eiff C., Kahl B. C., Becker K., McNamara P., Herrmann M. at al. Small colony variants: a pathogenic form of bacteria that facilitates persistent and recurrent infections. Nat Rev Microbiol 2006; 4 (Pt4): 295–305.Tuchscherr L., Medina E., Hussain M., Völker W., Heitmann V., Niemann ót al. Staphylococcus aureus phenotype switching: an effective bacterial strategy to escape host immune response and establish a chronic infection. EMBO Mol Med 2011; 3 (3): 129–141.Tuchscherr L., Medina E., Hussain M., Völker W., Heitmann V., Niemann ót al. Staphylococcus aureus phenotype switching: an effective bacterial strategy to escape host immune response and establish a chronic infection. EMBO Mol Med 2011; 3 (3): 129–141.Kahl B. C, Becker K., Löffler B. Clinical significance and pathogenesis of staphylococcal small colony variants in persistent infections. Clin Microbiol Rev 2016; 29 (2): 401–427.Kahl B. C, Becker K., Löffler B. Clinical significance and pathogenesis of staphylococcal small colony variants in persistent infections. Clin Microbiol Rev 2016; 29 (2): 401–427.Zhang P., Wright J. A., Osman A. A., Nair S. P. An aroD ochre mutation results in a Staphylococcus aureus small colony variant that can undergo phenotypic switching via two alternative mechanisms. Frontiers in Microbiology 2017; 8: 1001.Zhang P., Wright J. A., Osman A. A., Nair S. P. An aroD ochre mutation results in a Staphylococcus aureus small colony variant that can undergo phenotypic switching via two alternative mechanisms. Frontiers in Microbiology 2017; 8: 1001.Performance Standards for Antimicrobial Susceptibility Testing; Twenty First Informational Supplement, CLSI document M100-S24. Wayne, PA: Clinical and Laboratory Standards Institute: 2014.Performance Standards for Antimicrobial Susceptibility Testing; Twenty First Informational Supplement, CLSI document M100-S24. Wayne, PA: Clinical and Laboratory Standards Institute: 2014.European Committee on Antimicrobial Susceptibility Testing (EUCAST). Breakpoint tables for interpretation of MICs and zone diameters: Version 8.1. valid from 2018-05-15. (http://www.eucast.org).European Committee on Antimicrobial Susceptibility Testing (EUCAST). Breakpoint tables for interpretation of MICs and zone diameters: Version 8.1. valid from 2018-05-15. (http://www.eucast.org).Скала Л. 3., Лукин И.Н. Автоматизированное рабочее место микробиолога и химиотерапевта «Микроб-Автомат». Программное обеспечение. Версия 1.13, 2002-2009. Руководство пользователя. М.: Мед Проект-3, 2012. - 55 с.Skala L. Z., Lukin I.N. Avtomatizirovannoe rabochee mesto mikrobiologa i khimioterapevta «Mikrob-Avtomat». Programmnoe obespechenie. Versiya 1.13, 2002–2009. Rukovodstvo pol'zovatelya. M.: MedProekt-3, 2012; 55. [in Russian]Trukhacheva N.V. Matematicheskaya statistika v mediko-biologicheskikh issledovaniyakh s primeneniem paketa Statistica: M.: Geotar-Media; 2012. [in Russian]Trukhacheva N.V. Matematicheskaya statistika v mediko-biologicheskikh issledovaniyakh s primeneniem paketa Statistica: M.: Geotar-Media; 2012. [in Russian]Pu Y., Zhao Z., Li Y., Zou J., Ma Q., Zhao Y. et al. Enhanced efflux activity facilitates drug tolerance in dormant bacterial cells. Molecular cell 2016; 62 (2): 284–294.Pu Y., Zhao Z., Li Y., Zou J., Ma Q., Zhao Y. et al. Enhanced efflux activity facilitates drug tolerance in dormant bacterial cells. Molecular cell 2016; 62 (2): 284–294.Helaine S., Kugelberg E. Bacterial persisters: formation, eradication, and experimental systems. Trends in microbiology 2014; 22 (7): 417–424.Helaine S., Kugelberg E. Bacterial persisters: formation, eradication, and experimental systems. Trends in microbiology 2014; 22 (7): 417–424.Hofsteenge N., Van Nimwegen E., Silander O. K. Quantitative analysis of persister fractions suggests different mechanisms of formation among environmental isolates of E.coli. BMC microbiology 2013; 13 (1): 25.Hofsteenge N., Van Nimwegen E., Silander O. K. Quantitative analysis of persister fractions suggests different mechanisms of formation among environmental isolates of E.coli. BMC microbiology 2013; 13 (1): 25.Goneau L.W, Yeoh N. S, MacDonald K. W., Cadieux P. A., Burton J. P., Razvi H. et al. Selective target inactivation rather than global metabolic dormancy causes antibiotic tolerance in uropathogens. Antimicrob Agents Chemother 2014; AAC-02552.Goneau L.W, Yeoh N. S, MacDonald K. W., Cadieux P. A., Burton J. P., Razvi H. et al. Selective target inactivation rather than global metabolic dormancy causes antibiotic tolerance in uropathogens. Antimicrob Agents Chemother 2014; AAC-02552.Vulin C., Leimer N., Huemer M., Ackermann M., Zinkernagel A. S. Prolonged bacterial lag time results in small colony variants that represent a sub-population of persisters. Nature Comm 2018; 9 (1): 4074.Vulin C., Leimer N., Huemer M., Ackermann M., Zinkernagel A. S. Prolonged bacterial lag time results in small colony variants that represent a sub-population of persisters. Nature Comm 2018; 9 (1): 4074.Neou E., Michail G., Tsakris A., Pournaras S. Virulence of Acinetobacter baumannii Exhibiting Phenotypic Heterogeneous Growth against Meropenem in a Murine Thigh Infection Model. Antibiotics 2013; 2 (1): 73–82.Neou E., Michail G., Tsakris A., Pournaras S. Virulence of Acinetobacter baumannii Exhibiting Phenotypic Heterogeneous Growth against Meropenem in a Murine Thigh Infection Model. Antibiotics 2013; 2 (1): 73–82.

The authors declare that there are no conflicts of interest present.