Skip to main content

Advertisement

SpringerLink
Log in

Reactive oxygen species and uspA overexpession: an alternative bacterial response toward selection and maintenance of multidrug resistance in clinical isolates of uropathogenic E. coli

  • Original Article
  • Published:
European Journal of Clinical Microbiology & Infectious Diseases Aims and scope Submit manuscript

Abstract

Emergence of multidrug resistance (MDR) in uropathogenic E. coli (UPEC) demands alternative therapeutic interventions. Bactericidal antibiotics at their sub-inhibitory concentration stimulate production of reactive oxygen species (ROS) that results in oxidative stress, generates mutations, and alters transcription of different genes. Sub-inhibitory concentration of antibiotics facilitates selection of highly resistant population. Universal stress protein A (uspA) overexpression in MDR-UPEC at sub-inhibitory bactericidal antibiotics concentration was investigated to explore alternative survival strategy against them. Fifty clinical UPEC isolates were screened. Minimum inhibitory concentration (MIC) against three different bactericidal antibiotics (ciprofloxacin, CIP; ceftazidime, CAZ; gentamycin, GEN) was determined by broth dilution method; ROS production by DCFDA and overexpression of uspA by real-time PCR were determined at the sub-inhibitory concentration of antibiotics. DNA ladder formation and SEM studies were performed with drug untreated and treated samples. Statistical analysis was done by Student’s t test and Pearson’s correlation analysis; 25 out of 50 UPEC exhibited high MIC against CIP (> 200 μg/ml), CAZ (> 500 μg/ml), GEN (> 500 μg/ml), with varied ROS production (p ≤ 0.001) in treated than untreated controls. DNA ladder formation confirmed ROS production in drug-treated samples. SEM analysis revealed unaltered cell morphology in both untreated and drug-treated bacteria. uspA was universally overexpressed in all 25 UPEC. A significant correlation (p ≤ 0.001) between ROS production and uspA overexpression was observed in 19 out of 25 MDR isolates at sub-inhibitory doses of the bactericidal antibiotics. Therefore, this study highlights an alternative strategy that the MDR isolates may acquire when exposed to sub-inhibitory drug concentration for their survival.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Akram M, Shahid M, Khan A (2007) Etiology and antibiotic resistance patterns of community-acquired urinary tract infections in JNMC Hospital Aligarh, India. Ann Clin Microbiol Antimicrob 6 . https://doi.org/10.1186/1476-0711-6-4

  2. Moura A, Nicolau A, Hooton T, Azeredo J (2009) Antibiotherapy and pathogenesis of uncomplicated UTI: difficult relationships. J Appl Microbiol 106:1779–1791

    Article  CAS  Google Scholar 

  3. Gupta K, Hooton TM, Naber KG, Björn W, Colgan R, Miller LG, Moran GJ, Nicolle LE, Raz R, Schaeffer AJ (2011) Highlights from international clinical practice guidelines for the treatment of acute uncomplicated cystitis and pyelonephritis in women: a 2010 update by the infectious diseases society of America and the european society for microbiology and infectious. Infect Dis Clin Pract 19:282–283

    Article  Google Scholar 

  4. Hoban DJ, Nicolle LE, Hawser S, Bouchillon S, Badal R (2011) Antimicrobial susceptibility of global inpatient urinary tract isolates of Escherichia coli: results from the study for monitoring antimicrobial resistance trends (SMART) program: 2009-2010. Diagn Microbiol Infect Dis 70:507–511. https://doi.org/10.1016/j.diagmicrobio.2011.03.021

    Article  PubMed  Google Scholar 

  5. Bidell MR, Palchak M, Mohr J, Lodise TP (2016) Fluoroquinolone and third-generation-cephalosporin resistance among hospitalized patients with urinary tract infections due to Escherichia coli: Do rates vary by hospital characteristics and geographic region? Antimicrob Agents Chemother 60(3):170–173. https://doi.org/10.1128/aac.02505-15

    Article  Google Scholar 

  6. Tangcharoensathien V, Chanvatik S, Sommanustweechai A (2018) Complex determinants of inappropriate use of antibiotics. Bull World Health Organ 96:141–144. https://doi.org/10.2471/BLT.17.199687

    Article  PubMed  PubMed Central  Google Scholar 

  7. Kohanski MA, DePristo MA, Collins JJ (2010) Sub-lethal antibiotic treatment leads to multidrug resistance via radical-induced mutagenesis. Public Access NIH Public Access 12(37):311–320

    Google Scholar 

  8. Dridi B, Lupien A, Bergeron MG, Leprohon P, Ouellette M (2015) Differences in antibiotic-induced oxidative stress responses between laboratory and clinical isolates of streptococcus pneumoniae. Antimicrob Agents Chemother 59:5420–5426. https://doi.org/10.1128/AAC.00316-15

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Kvint K, Nachin L, Diez A, Nyström T (2003) The bacterial universal stress protein: function and regulation. Curr Opin Microbiol 6:140–145. https://doi.org/10.1016/S1369-5274(03)00025-0

    Article  CAS  PubMed  Google Scholar 

  10. Nachin L, Nannmark U, Nystro T (2005) Differential roles of the universal stress proteins of Escherichia coli in oxidative stress resistance, adhesion, and motility. J Bacteriol 187:6265–6272. https://doi.org/10.1128/JB.187.18.6265

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Weinstein MP, Patel JB, Bobenchik AM, Campeau S, Cullen SK, Galas MFGH (2019) Performance standards for antimicrobial susceptibility testing performance standards for antimicrobial susceptibility testing. CLSI, Suppl M100:1–25

    Google Scholar 

  12. Bandyopadhyay D, Mukherjee M (2017) Distribution of class d oxacillinases amongst third generation cephalosporin resistant nosocomial uropathogenic Escherichia coli isolates, their phylogenetic background and clonal analysis. Int J Cur Res 9:59099–59106

    Google Scholar 

  13. Bang CS, Kinnunen A, Karlsson M, Önnberg A, Söderquist B, Persson K (2014) The antibacterial effect of nitric oxide against ESBL-producing uropathogenic E. coli is improved by combination with miconazole and polymyxin B nonapeptide. BMC Microbiol 14:1–9. https://doi.org/10.1186/1471-2180-14-65

    Article  CAS  Google Scholar 

  14. Bhattacharya G, Dey D, Das S, Banerjee A (2017) Exposure to sub-inhibitory concentrations of gentamicin, ciprofloxacin and cefotaxime induces multidrug resistance and reactive oxygen species generation in meticillin-sensitive Staphylococcus aureus. J Med Microbiol 66:762–769

    Article  CAS  Google Scholar 

  15. Van Acker H, Gielis J, Acke M, Cools F, Cos P, Coenye T (2016) The role of reactive oxygen species in antibiotic-induced cell death in Burkholderia cepacia complex bacteria. PLoS One 11:1–20. https://doi.org/10.1371/journal.pone.0159837

    Article  CAS  Google Scholar 

  16. Ma L, Gao Y, Maresso AW (2015) Escherichia coli free radical-based killing mechanism driven by a unique combination of iron restriction and certain antibiotics. J Bacteriol 197:3708–3719. https://doi.org/10.1128/jb.00758-15

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. An SH, Kang JH. 2013 Oxidative damage of DNA induced by the reaction of methylglyoxal with lysine in the presence of ferritin. BMB Rep 46 (2013)225–229. https://doi.org/10.5483/BMBRep.46:225

  18. Seifart Gomes C, Izar B, Pazan F, Mohamed W, Mraheil MA, Mukherjee K et al (2011) Universal stress proteins are important for oxidative and acid stress resistance and growth of listeria monocytogenes EGD-e in vitro and in vivo. PLoS One 6:e24965. https://doi.org/10.1371/journal.pone.0024965

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Chauhan AK, Kang SC (2014) Thymol disrupts the membrane integrity of Salmonella ser. typhimurium in vitro and recovers infected macrophages from oxidative stress in an ex vivo model. Res Microbiol 165:559–565. https://doi.org/10.1016/j.resmic.2014.07.001

    Article  CAS  PubMed  Google Scholar 

  20. Baral P, Neupane S, Marasini B, Ghimire K, Lekhak B, Shrestha B (2012) High prevalence of multidrug resistance in bacterial uropathogens from Kathmandu. Nepal BMC Res Notes 5. https://doi.org/10.1186/1756-0500-5-38

  21. Taneja N, Rao P, Arora J, Dogra A (2008) Occurrence of ESBL & Amp-C b -lactamases & susceptibility to newer antimicrobial agents in complicated UTI. Indian J Med Res 127:85–88

    PubMed  Google Scholar 

  22. Norrby SR, Eriksson M, Ottosson EVA (1986) Imipenem/cilastatin versus gentamicin/clindamycin: a cost effectiveness study. Scand J Infect Dis 18:371–374

    Article  CAS  Google Scholar 

  23. McKinnon PS, Paladino JA, Grayson ML, Gibbons GW, Karchmer AW (1997) Cost-effectiveness of ampicillin/sulbactam versus imipenem/cilastatin in the treatment of limb-threatening foot infections in diabetic patients. Clin Infect Dis 24:57–63. https://doi.org/10.1093/clinids/24.1.57

    Article  CAS  PubMed  Google Scholar 

  24. Fortman JL, Mukhopadhyay A (2016) The future of antibiotics: emerging technologies and stewardship. Trends Microbiol 24:515–517. https://doi.org/10.1016/j.tim.2016.04.003

    Article  CAS  PubMed  Google Scholar 

  25. Gullberg E, Sandegren L, Hughes D, Dan I, Cao S, Berg OG et al (2011) Selection of resistant bacteria at very low antibiotic concentrations. PLoS Pathog 7:1–9. https://doi.org/10.1371/journal.ppat.1002158

    Article  CAS  Google Scholar 

  26. Wang P, Zhang X, Wang L, Zhen Z, Tang M, Li J (2010) Subinhibitory concentrations of ciprofloxacin induce SOS response and mutations of antibiotic resistance in bacteria. Ann Microbiol 60:511–517. https://doi.org/10.1007/s13213-010-0080-x

    Article  CAS  Google Scholar 

  27. Dwyer DJ, Belenky PA, Yang JH, Cody MacDonald I, Martell JD, Takahashi N et al (2014) Antibiotics induce redox-related physi. Proc Natl Acad Sci U S A:111. https://doi.org/10.1073/pnas.1401876111

  28. Keren I, Wu Y, Inocencio J, Mulcahy LR, Lewis K (2013) Killing by bactericidal antibiotics does not depend on reactive oxygen species. Science 339:1213–1216. https://doi.org/10.1126/science.1232688

    Article  CAS  PubMed  Google Scholar 

  29. Zhao X, Drlica K (2014) Reactive oxygen species and the bacterial response to lethal stress. Public Access NIH Public Access:1–6

  30. Dwyer DJ, Camacho DM, Kohanski MA, Callura JM, Collins JJ (2012) Antibiotic-induced bacterial cell death exhibits physiological and biochemical hallmarks of apoptosis. Mol Cell 46:61–72. https://doi.org/10.1016/j.molcel.2012.04.027

    Article  CAS  Google Scholar 

  31. Nyström T, Neidhardt FC (1996) Effects of overproducing the universal stress protein, UspA, in Escherichia coli K-12. J Bacteriol 178:927–930. https://doi.org/10.1128/jb.178.3.927-930.1996

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

We are grateful to Prof (Dr.) Pratip Kumar Kundu, Director, and Prof (Dr.) Bibhuti Saha Head, Department of Tropical Medicine, School of Tropical Medicine, Kolkata West Bengal, India, for their kind support.

Funding

This work was supported by a grant from the Department of Science and Technology, Government of West Bengal (Grant no. 755 (Sanc.)/ST/P/S&T/9G-26/2014 dated 17 December 2015).

Author information

Authors and Affiliations

  1. Department of Biochemistry and Medical Biotechnology, Calcutta School of Tropical Medicine, Kolkata, West Bengal, 700073, India

    Debojyoty Bandyopadhyay & Mandira Mukherjee

Authors
  1. Debojyoty Bandyopadhyay
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mandira Mukherjee
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mandira Mukherjee.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

The study protocol was approved by the Institutional Clinical Research Ethics Committee (CREC-STM/250 dated 9 January 2015).

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bandyopadhyay, D., Mukherjee, M. Reactive oxygen species and uspA overexpession: an alternative bacterial response toward selection and maintenance of multidrug resistance in clinical isolates of uropathogenic E. coli. Eur J Clin Microbiol Infect Dis 39, 1753–1760 (2020). https://doi.org/10.1007/s10096-020-03903-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10096-020-03903-x

Keywords

Search

Navigation