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Folia Microbiologica

, Volume 63, Issue 4, pp 451–457 | Cite as

Alterations of growth rate and gene expression levels of UPEC by antibiotics at sub-MIC

  • Defne Gümüş
  • Fatma Kalaycı-Yüksek
  • Emre Yörük
  • Gülşen Uz
  • Eşref Çelik
  • Cansu Arslan
  • Elif Merve Aydın
  • Cem Canlı
  • Mine Anğ-Küçüker
Original Article
  • 105 Downloads

Abstract

The host is the main environment for bacteria, and they also expose to many antibiotics during the treatment of infectious diseases in host body. In this study, it was aimed to investigate possible changes in growth rate and expression levels of three virulence genes (foc/foc, cnf1, and usp) in a uropathogenic E. coli standard strain within the presence of ciprofloxacin, nitrofurantoin, and trimethoprim-sulfamethoxazole. The UPEC C7 strain was grown on tryptic soy broth-TSB (control), TSB + ciprofloxacin, TSB + nitrofurantoin, and TSB + trimethoprim-sulfamethoxazole for determination of both growth rate and gene expression level. Antibiotics were added according to their sub-minimal inhibition concentrations. E-test was used to determine MIC values of antibiotics. Growth changes were measured in absorbance 600 nm during 24-h period. Total RNA isolations were performed after incubation for 24 h at 37 °C. Gene expression levels were determined by quantitative PCR. Tukey’s post hoc test was used for statistical analysis. According to absorbance values, it has been shown that only ciprofloxacin and trimethoprim-sulfamethoxazole have lead significant decrease on growth rate. We also detected statistically significant differences in each gene expression levels for all antibiotics via relative quantification analysis. Fold changes in gene expression was found 0.65, 1.42, 0.23 for foc/foc gene; 0.01, 0.01, 2.84 for cnf1 gene; and 0.1, 0.01, 0.01 for usp gene in the presence of ciprofloxacin, nitrofurantoin, and trimethoprim/sulfamethoxazole, respectively. This investigation has shown that antibiotics can play a role as an environmental factor which may determine the pathogenicity of bacteria in vivo.

Notes

Funding information

This research was supported by TUBITAK (The Scientific and Technological Research Council of Turkey) (Project No.: 2209-a / 2015) and Research Fund of Istanbul Yeni Yuzyil University.

Compliance with ethical standards

Conflict of interest

We would like to inform you that our manuscript has not been accepted for publication elsewhere. The authors of the article do not want to declare anything about their interest; and there is no conflict among the authors on this paper “Alterations of growth rate and gene expression levels of UPEC by antibiotics at sub-MIC.”

References

  1. Amini B, Baghchesaraie H, Haji Ojagh Faghihi M (2009) Effect of different sub MIC concentrations of penicillin, vancomycin and ceftazidime on morphology and some biochemical properties of Staphylococcus aureus and Pseudomonas aeruginosa isolates. Iran J Microbiol 1:43–47Google Scholar
  2. Andersson DI, Diarmaid H (2014) Microbiological effects of sublethal levels of antibiotics. Nat Rev Microbiol 12:465–478.  https://doi.org/10.1038/nrmicro3270 CrossRefPubMedGoogle Scholar
  3. Babić F, Venturi V, Maravić-Vlahoviček G (2010) Tobramycin at subinhibitory concentration inhibits the RhlI/R quorum sensing system in a Pseudomonas aeruginosa environmental isolate. BMC Infect Dis 10(1):148.  https://doi.org/10.1186/1471-2334-10-148 CrossRefPubMedPubMedCentralGoogle Scholar
  4. Balagué C, Fernández L, Pérez J, Grau R (2003) Effect of ciprofloxacin on adhesive properties of non-P mannose-resistant uropathogenic Escherichia coli isolates. J Antimicrob Chemother 51:401–404CrossRefPubMedGoogle Scholar
  5. Balaji K, Thenmozhi R, Pandian SK (2013) Effect of subinhibitory concentrations of fluoroquinolones on biofilm production by clinical isolates of Streptococcus pyogenes. Indian J Med Res 137:963PubMedPubMedCentralGoogle Scholar
  6. Baquero F, Tedim AP, Coque TM (2013) Antibiotic resistance shaping multi-level population biology of bacteria. Front Microbiol 4:15.  https://doi.org/10.3389/fmicb.2013.00015 CrossRefPubMedPubMedCentralGoogle Scholar
  7. Baskin H, Dogan Y, Bahar IH, Yulug N (2002) Effect of subminimal inhibitory concentrations of three fluoroquinolones on adherence of uropathogenic strains of Escherichia coli. Int J Antimicrob Agents 19:79–82CrossRefPubMedGoogle Scholar
  8. Bernier SP, Surette MG (2013) Concentration dependent activity of antibiotics in natural environments. Front Microbiol 4:20.  https://doi.org/10.3389/fmicb.2013.00020 CrossRefPubMedPubMedCentralGoogle Scholar
  9. Brunelle BW, Bearson SM, Bearson BL (2013) Tetracycline accelerates the temporally-regulated invasion response in specific isolates of multi drug resistant Salmonella enterica serovar Typhimurium. BMC Microbiol 13:202.  https://doi.org/10.1186/1471-2180-13-202 CrossRefPubMedPubMedCentralGoogle Scholar
  10. Calabrese EJ (2004) Hormesis: from marginalization to mainstream: a case for hormesis as the default dose-response model in risk assessment. Toxicol Appl Pharmacol 197:125–136.  https://doi.org/10.1016/j.taap.2004.02.007 CrossRefPubMedGoogle Scholar
  11. Dal Sasso M, Culici M, Bovio C, Braga PC (2003) Gemifloxacin: effects of sub-inhibitory concentrations on various factors affecting bacterial virulence. Int J Antimicrob Agents 21:325–333CrossRefPubMedGoogle Scholar
  12. Davies J, Spiegelman GB, Yim G (2006) The world of subinhibitory antibiotic concentrations. Curr Opin Microbiol 9:445–453.  https://doi.org/10.1016/j.mib.2006.08.006 CrossRefPubMedGoogle Scholar
  13. de Andrade JP, de Macêdo Farias L, Ferreira JF, Bruna-Romero O, da Glória de Souza D, de Carvalho MA, dos Santos KV (2016) Sub-inhibitory concentration of piperacillin–tazobactam may be related to virulence properties of filamentous Escherichia coli. Curr Microbiol 72:19–28.  https://doi.org/10.1007/s00284-015-0912-9 CrossRefPubMedGoogle Scholar
  14. Dhabaan GN, AbuBakar S, Cerqueira GM, Al-Haroni M, Pang SP, Hassan H (2016) Imipenem treatment induces expression of important genes and phenotypes in a resistant Acinetobacter baumannii isolate. Antimicrob Agents Chemothe 60:1370–1376.  https://doi.org/10.1128/AAC.01696-15 CrossRefGoogle Scholar
  15. Drummond Lisa J, Smith DGE, Poxton IR (2003) Effects of sub-MIC concentrations of antibiotics on growth of and toxin production by Clostridium difficile. J Med Microbiol 52(12):1033–1038.  https://doi.org/10.1099/jmm.0.05387-0 CrossRefPubMedGoogle Scholar
  16. Fajardo A, Martínez J (2008) Antibiotics as signals that trigger specific bacterial responses. Curr Opin Microbiol 11:161–167.  https://doi.org/10.1016/j.mib.2008.02.006 CrossRefPubMedGoogle Scholar
  17. Ferrer MD, Rodriguez JC, Álvarez L, Artacho A, Royo G, Mira A (2017) Effect of antibiotics on biofilm inhibition and induction measured by real-time cell analysis. J Appl Microbiol 122:640–650.  https://doi.org/10.1111/jam.13368 CrossRefPubMedGoogle Scholar
  18. Goh E, Yim G, Wayne T, McClure J, Surette MG, Davies J (2002) Transcriptional modulation of bacterial gene expression by subinhibitory concentrations of antibiotics. PNAS 99:17025–17030.  https://doi.org/10.1073/pnas.252607699 CrossRefPubMedPubMedCentralGoogle Scholar
  19. Haas B, Grenier D (2016) Impact of sub-inhibitory concentrations of amoxicillin on Streptococcus suis capsule gene expression and inflammatory potential. Pathogens 5:37.  https://doi.org/10.3390/pathogens5020037 CrossRefPubMedCentralGoogle Scholar
  20. Horii T, Morita M, Muramatsu H, Muranaka Y, Kanno T, Maekawa M (2003) Effects of mupirocin at subinhibitory concentrations on flagella formation in Pseudomonas aeruginosa and Proteus mirabilis. J Antimicrob Chemother 51:1175–1179.  https://doi.org/10.1093/jac/dkg226 CrossRefPubMedGoogle Scholar
  21. Ismaeel AY, Senok AC, Bindayna KM, Bakhiet M, Al Mahmeed A, Yousifa AQ, Bottam GA (2005) Effect of antibiotic subinhibitory concentration on cytolethal distending toxin production by Campylobacter jejuni. J Inf Secur 51:144–149.  https://doi.org/10.1016/j.jinf.2004.09.012 Google Scholar
  22. Kaper JB, Nataro JP, Mobley HL (2004) Pathogenic Escherichia coli. Nat Rev Microbiol 2:123–140.  https://doi.org/10.1038/nrmicro818 CrossRefPubMedGoogle Scholar
  23. Laureti L, Matic I, Gutierrez A (2013) Bacterial responses and genome instability induced by subinhibitory concentrations of antibiotics. Antibiotics 2:100–114.  https://doi.org/10.3390/antibiotics2010100 CrossRefPubMedPubMedCentralGoogle Scholar
  24. Linares JF, Gustafsson I, Baquero F, Martinez JL (2006) Antibiotics as intermicrobial signaling agents instead of weapons. Proc Natl Acad Sci U S A 103:19484–19489.  https://doi.org/10.1073/pnas.0608949103 CrossRefPubMedPubMedCentralGoogle Scholar
  25. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-Delta Delta C(T) method. Methods 25:402–408.  https://doi.org/10.1006/meth.2001.1262 CrossRefPubMedGoogle Scholar
  26. Majtan J, Majtanova L, Xu M, Majtan V (2007) In vitro effect of subinhibitory concentrations of antibiotics on biofilm formation by clinical strains of Salmonella enterica serovar Typhimurium isolated in Slovakia. J Appl Microbiol 104:1294–1301.  https://doi.org/10.1111/j.1365-2672.2007.03653.x CrossRefPubMedGoogle Scholar
  27. McKenney D, Willcock L, Trueman PA, Allison DG (1994) Effect of sub-MIC antibiotics on the cell surface and extracellular virulence determinants of Pseudomonas cepacia. J Appl Bacteriol 76:190–195CrossRefPubMedGoogle Scholar
  28. Molina-Quiroz RC, Silva CA, Molina CF, Leiva LE, Reyes-Cerpa S, Contreras I, Santiviago CA (2015) Exposure to sub-inhibitory concentrations of cefotaxime enhances the systemic colonization of Salmonella Typhimurium in BALB/c. Open Biol 5:150070.  https://doi.org/10.1098/rsob.150070 CrossRefPubMedPubMedCentralGoogle Scholar
  29. Mortensen NP, Fowlkes JD, Maggart M, Doktycz MJ, Nataro JP, Drusano G, Allison DP (2011) Effects of sub-minimum inhibitory concentrations of ciprofloxacin on enteroaggregative Escherichia coli and the role of the surface protein dispersin. Int J Antimicrob Agents 38:27–34.  https://doi.org/10.1016/j.ijantimicag.2011.03.011 CrossRefPubMedGoogle Scholar
  30. Moura TMD, Campos FS, Caierão J, Franco AC, Roehe PM, d’Azevedo PA et al (2015) Influence of a subinhibitory concentration of vancomycin on the in vitro expression of virulence-related genes in the vancomycin-resistant Enterococcus faecalis. Rev Soc Bras Med Trop 48:617–621.  https://doi.org/10.1590/0037-8682-0017-2015 CrossRefPubMedGoogle Scholar
  31. Nassar FJ, Rahal EA, Sabra A, Matar GM (2013) Effects of subinhibitory concentrations of antimicrobial agents on Escherichia coli O157: H7 Shiga toxin release and role of the SOS response. Foodborne Pathog Dis 10:805–812.  https://doi.org/10.1089/fpd.2013.1510 CrossRefPubMedGoogle Scholar
  32. Otto MP, Martin E, Badiou C, Lebrun S, Bes M, Vandenesch F, Dumitrescu O (2013) Effects of subinhibitory concentrations of antibiotics on virulence factor expression by community-acquired methicillin-resistant Staphylococcus aureus. J Antimicrob Chemother 68:1524–1532.  https://doi.org/10.1093/jac/dkt073 CrossRefPubMedGoogle Scholar
  33. Shen L, Shi Y, Zhang D, Wei J, Surette MG, Duan K (2008) Modulation of secreted virulence factor genes by subinhibitory concentrations of antibiotics in Pseudomonas aeruginosa. J Microbiol 46:441–447.  https://doi.org/10.1007/s12275-008-0054-x CrossRefPubMedGoogle Scholar
  34. Soto SM, De Anta MJ, Vila J (2006) Quinolones induce partial or total loss of pathogenicity islands in uropathogenic Escherichia coli by SOS-dependent or-independent pathways, respectively. Antimicrob Agents Chemother 50:649–653.  https://doi.org/10.1128/AAC.50.2.649-653.2006 CrossRefPubMedPubMedCentralGoogle Scholar
  35. Stamm WE (2006) Theodore E Woodward award: host-pathogen interactions in community-acquired urinary tract infections. Trans Am Clin Climatol Assoc 117:75–83PubMedPubMedCentralGoogle Scholar
  36. Subrt N, Mesak LR, Davies J (2011) Modulation of virulence gene expression by cell wall active antibiotics in Staphylococcus aureus. J Antimicrob Chemother 66:979–984.  https://doi.org/10.1093/jac/dkr043 CrossRefPubMedGoogle Scholar
  37. Talan DA, Krishnadasan A, Abrahamian FM, Stamm WE et al (2008) Prevalence and risk factor analysis of trimethoprimsulfamethoxazole—and fluoroquinolone-resistant Escherichia coli infection among emergency department patients with pyelonephritis. Clin Infect Dis 47:1150–1158.  https://doi.org/10.1086/592250 CrossRefPubMedGoogle Scholar
  38. Tateda K, Comte R, Pechere JC, Kohler T, Yamaguchi K, Delden CV (2001) Azithromycin inhibits quorum sensing in Pseudomonas aeruginosa. Antimicrob Agents Chemother 45:1930–1933.  https://doi.org/10.1128/AAC.45.6.1930-1933.2001 CrossRefPubMedPubMedCentralGoogle Scholar
  39. Tateda K, Ishii Y, Kimura S et al (2007) Suppression of Pseudomonas aeruginosa quorum-sensing systems by macrolides: a promising strategy or an oriental mystery? J Infect Chemother 13:357–367.  https://doi.org/10.1007/s10156-007-0555-2 CrossRefPubMedGoogle Scholar
  40. Vidya KC, Mallya PS, Rao PS (2005) Inhibition of bacterial adhesion by subinhibitory concentrations of antibiotics. Indian J Med Microbiol 23:102CrossRefPubMedGoogle Scholar
  41. Vranes J, Zagar Z, Kurbel S (1996) Influence of subinhibitory concentrations of ceftazidime, ciprofloxacin and azithromycin on the morphology and adherence of P-fimbriated Escherichia coli. J Chemother 8:254–260.  https://doi.org/10.1179/joc.1996.8.4.254 CrossRefPubMedGoogle Scholar
  42. Wayne PA (2012) Performance standards for antimicrobial susceptibility testing. Twenty second informational supplement update. CLSI document M100-S22 U Clinical and Laboratory Standards Institute, WayneGoogle Scholar
  43. Weir EK, Martin LC, Poppe C, Coombes BK, Boerlin P (2008) Subinhibitory concentrations of tetracycline affect virulence gene expression in a multiresistant Salmonella enterica subsp. enterica serovar Typhimurium DT104. Microbes Infect 10:901–907.  https://doi.org/10.1016/j.micinf.2008.05.005 CrossRefPubMedGoogle Scholar
  44. Wojnicz D, Cisowska A (2009) Composition of the outer membrane proteins of Escherichia coli strains in relation to serum susceptibility after exposure to subinhibitory concentrations of amikacin and ciprofloxacin. Int J Antimicrob Agents 33:579–582.  https://doi.org/10.1016/j.ijantimicag.2008.12.006 CrossRefPubMedGoogle Scholar
  45. Wojnicz D, Kłak M, Adamski R, Jankowski S (2007) Influence of subinhibitory concentrations of amikacin and ciprofloxacin on morphology and adherence ability of uropathogenic strains. Folia Microbiol (Praha) 52:429–436CrossRefGoogle Scholar
  46. Yim G, Wang HH, Davies J (2006) The truth about antibiotics. Int J Med Microbiol 296:163–170.  https://doi.org/10.1016/j.ijmm.2006.01.039 CrossRefPubMedGoogle Scholar
  47. Yim G, Wang HH, FRS JD (2007) Antibiotics as signalling molecules. Philos Trans R Soc Lond Ser B Biol Sci 362:1195–1200.  https://doi.org/10.1098/rstb.2007.2044 CrossRefGoogle Scholar
  48. Yim G, McClure J, Surette MG, Davies JE (2011) Modulation of Salmonella gene expression by subinhibitory concentrations of quinolones. The Journal of antibiotics 64:73–78.  https://doi.org/10.1038/ja.2010.137 CrossRefPubMedGoogle Scholar
  49. Zhang L, Foxman B (2003) Molecular epidemiology of Escherichia coli mediated urinary tract infections. Front Biosci 8:235–244CrossRefGoogle Scholar

Copyright information

© Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i. 2018

Authors and Affiliations

  1. 1.Department of Medical Microbiology, Faculty of MedicineIstanbul Yeni Yuzyil UniversityIstanbulTurkey
  2. 2.Department of Molecular Biology and Genetics, Faculty of Arts & SciencesIstanbul Yeni Yuzyil UniversityIstanbulTurkey

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