2020 / 03

Next Paper
DOI: 10.24075/brsmu.2020.032

ORIGINAL RESEARCH

Colistin resistance of carbapenem-resistant Klebsiella pneumoniae strains: molecular mechanisms and bacterial fitness

Shamina OV1, Kryzhanovskaya OA1, Lazareva AV1, Alyabieva NM1, Mayanskiy NA2
About authors

1 National Medical Research Center for Children's Health, Moscow, Russia

2 Pirogov Russian National Research Medical University, Moscow, Russia

Correspondence should be addressed: Olga V. Shamina
Lomonosovskiy prospect, 2, str. 1, Moscow, 119991; ur.xobni@animahs.aglo

About paper

Funding: the study was supported by the Russian Science Foundation (Project ID 20-15-00235).

Acknowledgements: the authors thank Polikarpova SV of Filatov City Clinical Hospital № 15 and Karaseva OV of the Research Institute of Emergency Pediatric Surgery and Traumatology for K. pneumoniae isolates.

Author contributionThe increasing use of colistin in the clinic has led to the emergence and spread of colistin resistance. According to the literature, antibiotic resistance can have a metabolic cost, resulting in poor adaptation and survival, i.e. reduced bacterial fitness. The aim of this study was to investigate molecular mechanisms underlying resistance to colistin and their effect on the bacterial fitness of carbapenem-resistant (carba-R) strains of K. pneumoniae isolated from the patients of Moscow hospitals in 2012–2017. Of 159 analyzed carba-R isolates, 71 (45%) were resistant to colistin (minimum inhibitory concentration over 2 mg/L). By conducting Sanger sequencing, we were able to identify the mechanisms underlying colistin resistance in 26 (37%) isolates. Growth curves were constructed by measuring optical density at 600 nm wavelength for 15 hours. The competitive growth of colistin-resistant (col-R) K. pneumoniae isolates was assessed relative to the colistinsusceptible (col-S) isolate. Col-R and col-S cultures harvested in the exponential phase were combined at the ratio of 1:1, incubated in the Luria-Bertani medium and plated onto Luria-Bertani agar plates with 10 mg/L colistin and without it. The competition index was calculated as the ratio of grown col-R and col-S colonies. Resistance to colistin did not affect the growth kinetics of K. pneumoniae, but did reduce the competitive ability of the bacteria as compared to the col-S isolates. However, some col-R isolates were more competitive than the col-S strains of the same sequence type. Further research is needed to elucidate the effects of colistin resistance on bacterial fitness.: Shamina OV planned and conducted the study, analyzed the literature, analyzed and interpreted the obtained data, and wrote the manuscript; Kryzhanovskaya OA, Lazareva AV, Alyabieva NM planned and conducted the study; Mayanskiy NA planned, conducted and supervised the study, analyzed the literature, collected, analyzed and interpreted the obtained data, and wrote the manuscript.

Received: 2020-05-18 Accepted: 2020-06-02 Published online: 2020-06-12
|
  1. Suvorova MP, Yakovlev SV, Beloborodov VB, Basin EE, Eliseeva KV, Kovelenov SV. Rasprostranennost' i klinicheskoe znachenie nozokomial'nykh infektsiy v lechebnykh uchrezhdeniyakh Rossii: issledovanie ERGINI. Antibiotiki i khimioterapiya. 2016; 61 (5–6): 32–42. Russian.
  2. Sukhorukova MV, Edelstein MV, Skleenova EY, Ivanchik NV, Mikotina AV, Dekhnich AV et al. Antimicrobial resistance of nosocomial Enterobacteriaceae isolates in Russia: results of multicenter epidemiological study «MARATHON» 2013–2014. CMAC. 2017; 19 (1): 49–56.
  3. Sukhorukova MV, Edelstein MV, Skleenova EY, Ivanchik NV, Timokhova АV, Dekhnich AV et al. Antimicrobial resistance of nosocomial Enterobacteriaceae isolates in Russia: results of national multicenter surveillance study «MARATHON» 2011–2012. CMAC. 2014; 16 (4): 254–65.
  4. Lee CR, Lee JH, Park KS, Kim YB, Jeong BC, Lee SH. Global dissemination of carbapenemase-producing Klebsiella pneumoniae: Epidemiology, genetic context, treatment options, and detection methods. Front Microbiol. 2016; 7: 1–30.
  5. Wyres KL, Holt KE. Klebsiella pneumoniae Population Genomics and Antimicrobial-Resistant Clones. Trends Microbiol. 2016; 24 (12): 944–56.
  6. Izdebski R, Baraniak A, Zabicka D, Machulska M, Urbanowicz P, Fiett J, et al. Enterobacteriaceae producing OXA-48-like carbapenemases in Poland, 2013-January 2017. J Antimicrob Chemother. 2018; 73 (3): 620–25.
  7. Wyres KL, Hawkey J, Hetland MAK, Fostervold A, Wick RR, Judd LM, et al. Emergence and rapid global dissemination of CTX-M-15- associated Klebsiella pneumoniae strain ST307. J Antimicrob Chemother. 2019; 74 (3): 577–81.
  8. Ah YM, Kim AJ, Lee JY. Colistin resistance in Klebsiella pneumoniae. International Journal of Antimicrobial Agents. 2014; 44 (1): 8–15.
  9. Rojas LJ, Salim M, Cober E, Richter SS, Perez F, Salata RA et al. Colistin Resistance in Carbapenem-Resistant Klebsiella pneumoniae: Laboratory Detection and Impact on Mortality. Clin Infect Dis. 2017; 64 (6): 711–8.
  10. Poirel L, Aurelie J, Nordmann P. Polymyxins: Antibacterial Activity, Susceptibility Testing, and Resistance Mechanisms Encoded by Plasmids or Chromosomes. Clin Microbiol Rev. 2017; 30 (2): 557–96.
  11. Guo B, Abdelraouf K, Ledesma KR, Nikolaou M, Tam VH. Predicting bacterial fitness cost associated with drug resistance. J Antimicrob Chemother. 2012; 67 (4): 928–32.
  12. Ternent L, Dyson RJ, Krachler AM, Jabbari S. Bacterial fitness shapes the population dynamics of antibiotic-resistant and -susceptible bacteria in a model of combined antibiotic and anti-virulence treatment. J Theor Biol. 2015; 372: 1–11.
  13. Beceiro A, Moreno A, Fernandez N, Vallejo JA, Aranda J, Adler B, et al. Biological cost of different mechanisms of colistin resistance and their impact on virulence in Acinetobacter baumannii. Antimicrob Agents Chemother. 2014; 58 (1): 518–26.
  14. Choi MJ, Ko KS. Loss of hypermucoviscosity and increased fitness cost in colistin-resistant Klebsiella pneumoniae sequence type 23 strains. Antimicrob Agents Chemother. 2015; 59 (11): 6763–73.
  15. eucast.org [Internet]. European Committee on Antimicrobial Susceptibility Testing (EUCAST). Break- point tables for interpretation of MICs and zone diameters, version 8.0; c2018 [cited 2018 Dec 25]. Available from: http://www.eucast.org/ clinical_breakpoints/.
  16. Shamina OV, Kryzhanovskaya OA, Lazareva AV, Alyabieva NM, Polikarpova SV, Karaseva OV, et al. Emergence of a ST307 clone carrying a novel insertion element MITEKpn1 in the mgrB gene among carbapenem-resistant Klebsiella pneumoniae from Moscow, Russia. Int J Antimicrob Agents. 2020; 55 (2): 105850.
  17. Klebsiella pneumoniae MLST [baza dannyh]. Available from: http://www.pasteur.fr/mlst/. Russian.
  18. ISfinder database [Internet] [cited 2018 Nov 18]. Available from: http://www-is.biotoul.fr/is.html.
  19. Baron S, Hadjadj L, Rolain JM, Olaitan AO. Molecular mechanisms of polymyxin resistance: knowns and unknowns. Int J Antimicrob Agents. 2016; 48 (6): 583–91.
  20. Shamina OV, Kryzhanovskaya OA, Lazareva AV, Polikarpova SV, Karaseva OV, Chebotar IV et al. The comparison of methods for determination of colistin susceptibility in carbapenem-resistant Klebsiella pneumoniae. Klin Lab Diagn. 2018; 63 (10): 646–50.
  21. Ageevets VA. Molekulyarnaya kharakteristika produtsentov karbapenemaz semeystva Enterobacteriaceae, vydelennykh v Sankt-Peterburge [dissertatsiya]. Spb., 2016. Russian.
  22. Tansarli GS, Papaparaskevas J, Balaska M, Samarkos M, Pantazatou A, Markogiannakis A, et al. Colistin resistance in carbapenemase-producing Klebsiella pneumoniae bloodstream isolates: Evolution over 15 years and temporal association with colistin use by time series analysis. Int J Antimicrob Agents. 2018; 52 (3): 397–403.
  23. Feretzakis G, Loupelis E, Sakagianni A, Skarmoutsou N, Michelidou S, Velentza A, et al. A 2-year single-centre audit on antibiotic resistance of Pseudomonas aeruginosa, Acinetobacter baumannii and Klebsiella pneumoniae strains from an intensive care unit and other wards in a general public hospital in Greece. Antibiotics (Basel). 2019; 8 (2): 62.
  24. Cannatelli A, Santos-Lopez A, Giani T, Gonzalez-Zorn B, Rossolini GM. Polymyxin resistance caused by mgrB inactivation is not associated with significant biological cost in Klebsiella pneumoniae. Antimicrob Agents Chemother. 2015; 59 (5): 2898– 900.
  25. Moskowitz SM, Brannon MK, Dasgupta N, Pier M, Sgambati N, Miller AK, et al. PmrB mutations promote polymyxin resistance of Pseudomonas aeruginosa isolated from colistin-treated cystic fibrosis patients. Antimicrob Agents Chemother. 2012; 56 (2): 1019–30.
  26. Nang SC, Morris FC, McDonald MJ, Han ML, Wang J, Strugnell RA, et al. Fitness cost of mcr-1-mediated polymyxin resistance in Klebsiella pneumoniae. J Antimicrob Chemother. 2018; 73 (6): 1604–10.
  27. Marcusson LL, Frimodt-Møller N, Hughes D. Interplay in the selection of fluoroquinolone resistance and bacterial fitness. PLoS Pathog. 2009; 5 (8): e1000541.
  28. Mu X, Wang N, Li X, Shi K, Zhou Z, Yu Y, et al. The Effect of Colistin Resistance-Associated Mutations on the Fitness of Acinetobacter baumannii. Front Microbiol. 2016; 7: 1715.