Abstract
Pseudomonas aeruginosa is an opportunistic human pathogen, which can cause severe urinary tract infections (UTIs). Because of the high intrinsic antibiotic resistance of P. aeruginosa and its ability to develop new resistances during antibiotic treatment, these infections are difficult to eradicate. The antibiotic susceptibility of 32 P. aeruginosa isolates from acute and chronic UTIs were analysed under standardized conditions showing 19% multi-drug resistant strains. Furthermore, the antibiotic tolerance of two P. aeruginosa strains to ciprofloxacin and tobramycin was analysed under urinary tract-relevant conditions which considered nutrient composition, biofilm growth, growth phase, and oxygen concentration. These conditions significantly enhance the antibiotic tolerance of P. aeruginosa up to 6000-fold indicating an adaptation of the bacterium to the specific conditions present in the urinary tract. This reversible phenomenon is possibly due to the increased formation of persister cells and is based on iron limitation in artificial urine. The results suggest that the general high antibiotic resistance of P. aeruginosa urinary tract isolates together with the increasing tolerance of P. aeruginosa grown under urinary tract conditions decrease the efficiency of antibiotic treatment of UTIs.
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Abbreviations
- MDR:
-
Multi-drug resistant
- MIC:
-
Minimal inhibitory concentration
- UTI:
-
Urinary tract infection
- AUM:
-
Artificial urine medium
References
Anderl JN, Zahller J, Roe F, Stewart PS (2003) Role of nutrient limitation and stationary-phase existence in Klebsiella pneumoniae biofilm resistance to ampicillin and ciprofloxacin. Antimicrob Agents Chemother 47:1251–1256
Andrews JM (2001) Determination of minimum inhibitory concentrations. J Antimicrob Chemother 48(Suppl 1):5–16
Anwar H, Strap JL, Chen K, Costerton JW (1992) Dynamic interactions of biofilms of mucoid Pseudomonas aeruginosa with tobramycin and piperacillin. Antimicrob Agents Chemother 36:1208–1214
Borriello G, Werner E, Roe F, Kim AM, Ehrlich GD, Stewart PS (2004) Oxygen limitation contributes to antibiotic tolerance of Pseudomonas aeruginosa in biofilms. Antimicrob Agents Chemother 48:2659–2664
Breidenstein EB, Khaira BK, Wiegand I, Overhage J, Hancock RE (2008) Complex ciprofloxacin resistome revealed by screening a Pseudomonas aeruginosa mutant library for altered susceptibility. Antimicrob Agents Chemother 52:4486–4491
Brooks T, Keevil CW (1997) A simple artificial urine for the growth of urinary pathogens. Lett Appl Microbiol 24:203–206
Davis RL, Koup JR, Williams-Warren J, Weber A, Smith AL (1985) Pharmacokinetics of three oral formulations of ciprofloxacin. Antimicrob Agents Chemother 28:74–77
de Beer D, Stoodley P, Roe F, Lewandowski Z (1994) Effects of biofilm structures on oxygen distribution and mass transport. Biotechnol Bioeng 43:1131–1138
Defez C, Fabbro-Peray P, Bouziges N, Gouby A, Mahamat A, Daures JP, Sotto A (2004) Risk factors for multidrug-resistant Pseudomonas aeruginosa nosocomial infection. J Hosp Infect 57:209–216
Egberts J, Soederhuizen W (1996) Urine samples before dinner are preferable when studying changes in endogenous nitrate production under uncontrolled dietary conditions. Clin Chim Acta 254:141–148
Ellis G, Adatia I, Yazdanpanah M, Makela SK (1998) Nitrite and nitrate analyses: a clinical biochemistry perspective. Clin Biochem 31:195–220
Fung DK, Chan EW, Chin ML, Chan RC (2010) Delineation of a bacterial starvation stress response network which can mediate antibiotic tolerance development. Antimicrob Agents Chemother 54:1082–1093
Giamarellou H (2010) Multidrug-resistant Gram-negative bacteria: how to treat and for how long. Int J Antimicrob Agents 36(Suppl 2):S50–S54
Giannakopoulos X, Evangelou A, Kalfakakou V, Grammeniatis E, Papandropoulos I, Charalambopoulos K (1997) Human bladder urine oxygen content: implications for urinary tract diseases. Int Urol Nephrol 29:393–401
Gilbert P, Collier PJ, Brown MR (1990) Influence of growth rate on susceptibility to antimicrobial agents: biofilms, cell cycle, dormancy, and stringent response. Antimicrob Agents Chemother 34:1865–1868
Goto T, Nakame Y, Nishida M, Ohi Y (1999) Bacterial biofilms and catheters in experimental urinary tract infection. Int J Antimicrob Agents 11:227–231 discussion 237–239
Harmsen M, Yang L, Pamp SJ, Tolker-Nielsen T (2010) An update on Pseudomonas aeruginosa biofilm formation, tolerance, and dispersal. FEMS Immunol Med Microbiol 59:253–268
Hassett DJ, Cuppoletti J, Trapnell B, Lymar SV, Rowe JJ, Yoon SS, Hilliard GM, Parvatiyar K, Kamani MC, Wozniak DJ, Hwang SH, McDermott TR, Ochsner UA (2002) Anaerobic metabolism and quorum sensing by Pseudomonas aeruginosa biofilms in chronically infected cystic fibrosis airways: rethinking antibiotic treatment strategies and drug targets. Adv Drug Deliv Rev 54:1425–1443
Heydorn A, Ersboll BK, Hentzer M, Parsek MR, Givskov M, Molin S (2000) Experimental reproducibility in flow-chamber biofilms. Microbiology 146:2409–2415
Hoiby N, Bjarnsholt T, Givskov M, Molin S, Ciofu O (2010) Antibiotic resistance of bacterial biofilms. Int J Antimicrob Agents 35:322–332
Hryniewicz K, Szczypa K, Sulikowska A, Jankowski K, Betlejewska K, Hryniewicz W (2001) Antibiotic susceptibility of bacterial strains isolated from urinary tract infections in Poland. J Antimicrob Chemother 47:773–780
Kim J, Hahn JS, Franklin MJ, Stewart PS, Yoon J (2009) Tolerance of dormant and active cells in Pseudomonas aeruginosa PA01 biofilm to antimicrobial agents. J Antimicrob Chemother 63:129–135
Lambert PA (2002) Mechanisms of antibiotic resistance in Pseudomonas aeruginosa. J R Soc Med 95(Suppl 41):22–26
Lewis K (2007) Persister cells, dormancy and infectious disease. Nat Rev Microbiol 5:48–56
Mah TF, O’Toole GA (2001) Mechanisms of biofilm resistance to antimicrobial agents. Trends Microbiol 9:34–39
Mah TF, Pitts B, Pellock B, Walker GC, Stewart PS, O’Toole GA (2003) A genetic basis for Pseudomonas aeruginosa biofilm antibiotic resistance. Nature 426:306–310
Mittal R, Aggarwal S, Sharma S, Chhibber S, Harjai K (2009) Urinary tract infections caused by Pseudomonas aeruginosa: a minireview. J Infect Public Health 2:101–111
Mittal R, Sharma S, Chhibber S, Harjai K (2008) Iron dictates the virulence of Pseudomonas aeruginosa in urinary tract infections. J Biomed Sci 15:731–741
Nickel JC, Ruseska I, Wright JB, Costerton JW (1985) Tobramycin resistance of Pseudomonas aeruginosa cells growing as a biofilm on urinary catheter material. Antimicrob Agents Chemother 27:619–624
Obritsch MD, Fish DN, MacLaren R, Jung R (2004) National surveillance of antimicrobial resistance in Pseudomonas aeruginosa isolates obtained from intensive care unit patients from 1993 to 2002. Antimicrob Agents Chemother 48:4606–4610
Obritsch MD, Fish DN, MacLaren R, Jung R (2005) Nosocomial infections due to multidrug-resistant Pseudomonas aeruginosa: epidemiology and treatment options. Pharmacotherapy 25:1353–1364
Palmer KL, Brown SA, Whiteley M (2007) Membrane-bound nitrate reductase is required for anaerobic growth in cystic fibrosis sputum. J Bacteriol 189:4449–4455
Pascual A, Martinez–Martinez L, Ramirez de Arellano E, Perea EJ (1993) Susceptibility to antimicrobial agents of Pseudomonas aeruginosa attached to siliconized latex urinary catheters. Eur J Clin Microbiol Infect Dis 12:761–765
Plosker GL (2010) Aztreonam lysine for inhalation solution: in cystic fibrosis. Drugs 70:1843–1855
Raja NS, Singh NN (2007) Antimicrobial susceptibility pattern of clinical isolates of Pseudomonas aeruginosa in a tertiary care hospital. J Microbiol Immunol Infect 40:45–49
Rivera-Sanchez R, Delgado-Ochoa D, Flores-Paz RR, Garcia-Jimenez EE, Espinosa-Hernandez R, Bazan-Borges AA, Arriaga-Alba M (2010) Prospective study of urinary tract infection surveillance after kidney transplantation. BMC Infect Dis 10:245
Ronald A (2002) The etiology of urinary tract infection: traditional and emerging pathogens. Am J Med 113(Suppl 1A):14S–19S
Rossolini GM, Mantengoli E (2005) Treatment and control of severe infections caused by multiresistant Pseudomonas aeruginosa. Clin Microbiol Infect 11(Suppl 4):17–32
Saha R, Jain S, Kaur IR (2010) Metallo beta-lactamase producing pseudomonas species—a major cause of concern among hospital associated urinary tract infection. J Indian Med Assoc 108:344–348
Shigemura K, Arakawa S, Sakai Y, Kinoshita S, Tanaka K, Fujisawa M (2006) Complicated urinary tract infection caused by Pseudomonas aeruginosa in a single institution (1999–2003). Int J Urol 13:538–542
Silley P, Griffiths JW, Monsey D, Harris AM (1990) Mode of action of GR69153, a novel catechol-substituted cephalosporin, and its interaction with the tonB-dependent iron transport system. Antimicrob Agents Chemother 34:1806–1808
Spoering AL, Lewis K (2001) Biofilms and planktonic cells of Pseudomonas aeruginosa have similar resistance to killing by antimicrobials. J Bacteriol 183:6746–6751
Stewart PS, Franklin MJ (2008) Physiological heterogeneity in biofilms. Nat Rev Microbiol 6:199–210
Tenke P, Kovacs B, Jackel M, Nagy E (2006) The role of biofilm infection in urology. World J Urol 24:13–20
Tielen P, Narten M, Rosin N, Biegler I, Haddad I, Hogardt M, Neubauer R, Schobert M, Wiehlmann L, Jahn D (2010) Genotypic and phenotypic characterization of Pseudomonas aeruginosa isolates from urinary tract infections. Int J Med Microbiol 301:282–292
Tielen P, Rosenau F, Wilhelm S, Jaeger KE, Flemming HC, Wingender J (2010) Extracellular enzymes affect biofilm formation of mucoid Pseudomonas aeruginosa. Microbiol 156:2239–2252
Tuomanen E, Durack DT, Tomasz A (1986) Antibiotic tolerance among clinical isolates of bacteria. Antimicrob Agents Chemother 30:521–527
van Delden C (2004) Virulence factors in Pseudomonas aeruginosa. In: Ramos JL (ed) Pseudomonas. Kluwer Academics/Plenum Publisher, New York, pp 3–46
van Poppel H, Chysky V, Hullmann R, Baert L (1988) Clinical experience with ciprofloxacin in the treatment of urinary tract infections: a review. Infection 16:337–344
Waite RD, Papakonstantinopoulou A, Littler E, Curtis MA (2005) Transcriptome analysis of Pseudomonas aeruginosa growth: comparison of gene expression in planktonic cultures and developing and mature biofilms. J Bacteriol 187:6571–6576
Wang J, Brown MA, Tam SH, Chan MC, Whitworth JA (1997) Effects of diet on measurement of nitric oxide metabolites. Clin Exp Pharmacol Physiol 24:418–420
Werner E, Roe F, Bugnicourt A, Franklin MJ, Heydorn A, Molin S, Pitts B, Stewart PS (2004) Stratified growth in Pseudomonas aeruginosa biofilms. Appl Environ Microbiol 70:6188–6196
Zhao Z, Ma Y, Dai C, Zhao R, Li S, Wu Y, Cao Z, Li W (2009) Imcroporin, a new cationic antimicrobial peptide from the venom of the scorpion Isometrus maculates. Antimicrob Agents Chemother 53:3472–3477
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This work was supported by the BMBF GenoMik-Plus (Fkz: 0313801H).
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Narten, M., Rosin, N., Schobert, M. et al. Susceptibility of Pseudomonas aeruginosa Urinary Tract Isolates and Influence of Urinary Tract Conditions on Antibiotic Tolerance. Curr Microbiol 64, 7–16 (2012). https://doi.org/10.1007/s00284-011-0026-y
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DOI: https://doi.org/10.1007/s00284-011-0026-y