Advertisement

Applied Microbiology and Biotechnology

, Volume 81, Issue 2, pp 349–358 | Cite as

Investigating the effect of patulin, penicillic acid and EDTA on biofilm formation of isolates from dental unit water lines

  • Iram Liaqat
  • Robert Th. Bachmann
  • Anjum Nasim Sabri
  • Robert G. J. Edyvean
  • Catherine A. Biggs
Applied Microbial and Cell Physiology

Abstract

This study investigated the effect of patulin and penicillic acid, two known quorum-sensing inhibitors, and the common biocide ethylenediaminetetraacetic acid (EDTA) on the biofilm formation and auto-inducer (AI)-2 production of three isolates from dental unit water lines, Klebsiella sp., Bacillus subtilis and Bacillus cereus. Penicillic acid on its own had no effect on the biofilm formation of all isolates, whereas in combination with EDTA, it enhanced biofilm formation significantly in Klebsiella sp. and B. cereus. EDTA at concentrations greater than 10 μM promoted biofilm formation in B. cereus and B. subtilis. Patulin was found to promote biofilm formation in B. cereus up to 25 μM. A significant increase in biofilm formation was observed in B. cereus and B. subtilis at concentrations greater than 10 μM of patulin when combined with EDTA. The Vibrio harveyi BB170 AI-2 bioassay showed a positive response for Klebsiella sp. AI-2 production with a maximum fold induction at the late exponential growth phase. Addition of glucose prolonged the AI-2 production phase considerably. No significant effect of patulin, penicillic acid alone as well as in combination with EDTA was observed on AI-2 production by Klebsiella sp. The findings have important implications for the design of biofilm prevention and eradication strategies.

Keywords

DUWL isolates Patulin Penicillic acid Biocide Biofilm assay AI-2 assay 

Notes

Acknowledgements

The authors wish to thank Kevin Eboigbodin, Ammar Abdul Razak, Ernesto Hernandez and Johanna Andrews for their helpful discussion. We thank Michel Gohar for his timely guidance regarding our project. This work was funded by Higher Education Commission (HEC), Pakistan. The authors also wish to thank Professor Paul Williams (University of Nottingham, UK) for the V. harveyi strains for the AI-2 bioassay. Author Catherine Biggs also wishes to thank the Engineering and Physical Sciences Research Council UK for an advanced research fellowship (EP/E053556/1).

Supplementary material

253_2008_1691_MOESM1_ESM.doc (30 kb)
Supplementary Material (DOC 30 KB)

References

  1. Auger S, Krin E, Aymerich S, Gohar M (2006) Autoinducer 2 affects biofilm formation by Bacillus cereus. Appl Environ Microbiol 72:937–941CrossRefGoogle Scholar
  2. Balestrino D, Haagensen JA, Rich C, Forestier C (2005) Characterization of type 2 quorum sensing in Klebsiella pneumoniae and relationship with biofilm formation. J Bacteriol 187:2870–2880CrossRefGoogle Scholar
  3. Banin E, Brady KM, Greenberg EP (2006) Chelator-induced dispersal and killing of Pseudomonas aeruginosa cells in a biofilm. Appl Environ Microbiol 72:2064–2069CrossRefGoogle Scholar
  4. Barak I, Ricca E, Cutting S (2005) From fundamental studies of sporulation to applied spore research. Mol Microbiol 55:330–338CrossRefGoogle Scholar
  5. Bassler BL, Greenberg EP, Stevens AM (1997) Cross-species induction of luminescence in the quorum-sensing bacterium Vibrio harveyi. J Bacteriol 179:4043–4045Google Scholar
  6. Branda SS, Vik S, Friedman L, Kolter R (2006) Biofilms: the matrix revisited. Trends Microbiol 13:20–26CrossRefGoogle Scholar
  7. Burton E, Yakandawala N, LoVetri K, Madhyastha MS (2007) A microplate spectrofluorometric assay for bacterial biofilms. J Ind Microbiol Biotechnol 34:1–4CrossRefGoogle Scholar
  8. Christensen GD, Simpson WA, Younge J, Baddour LM, Barrett FF, Melton DM, Beachey EH (1985) Adherence of coagulasenegative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol 22:996–1006Google Scholar
  9. Decho AW (1990) Microbial exopolymer secretions in ocean environments: their role(s) in food webs and marine processes. Oceanogr Mar Bio Annu Rev 28:73–153Google Scholar
  10. Eboigbodin KE, Newton JRA, Routh AF, Biggs CA (2006) Bacterial quorum sensing and cell surface electrokinetic properties. Appl Microbiol Biotechnol 73:669–675CrossRefGoogle Scholar
  11. Frezza M, Soulère L, Balestrino D, Gohar M, Deshayes C, Queneau Y, Forestier C, Doutheau A (2007) Ac2-DPD, the bis-(O)-acetylated derivative of 4,5-dihydroxy-2,3-pentanedione (DPD) is a convenient stable precursor of bacterial quorum sensing autoinducer AI-2. Bioorg Med Chem Lett 17:1428–1431CrossRefGoogle Scholar
  12. Hansen LT, Austin JW, Gill TA (2001) Antibacterial effect of protamine in combination with EDTA and refrigeration. Int J Food Microbiol 66:149–161CrossRefGoogle Scholar
  13. Hentzer M, Givskov M (2003) Pharmacological inhibition of quorum sensing for the treatment of chronic bacterial infections. J Clin Invest 112:1300–1307Google Scholar
  14. Hentzer M, Riedel K, Rasmussen TB, Heydorn A, Andersen JB, Parsek MR, Rice SA, Eberl L, Moli S, Hoiby N, Kjelleberg S, Givskov M (2002) Inhibition of quorum sensing in Pseudomonas aeruginosa biofilm bacteria by a halogenated furanone compound. Microbiology 148:87–102Google Scholar
  15. Jones MB, Blaser MJ (2003) Detection of a luxS-signaling molecule in Bacillus anthracis. Infect Immun 71:914–3919Google Scholar
  16. Liaqat I, Sabri AN (2008a) Analysis of cell wall constituents of biocides resistant isolates from dental unit water line biofilms. Curr Microbiol 57:340–347CrossRefGoogle Scholar
  17. Liaqat I, Sabri AN (2008b) Effect of biocides on biofilm bacteria from dental unit water lines. Curr Microbiol 56:619–624CrossRefGoogle Scholar
  18. Liu H, Fang HP (2002) Characterization of electrostatic binding sites of extracellular polymers by linear programming analysis of titration data. Biotechnol Bioeng 80:806–811CrossRefGoogle Scholar
  19. Lombardía E, Rovetto AJ, Arabolaza AL, Grau RR (2006) A LuxS-dependent cell-to-cell language regulates social behavior and development in Bacillus subtilis. J Bacteriol 188:4442–4452CrossRefGoogle Scholar
  20. Olofsson AC, Hermansson M, Elwing H (2003) N-acetyl-L-cysteine affects growth, extracellular polysaccharide production, and bacterial biofilm formation on solid surfaces. Appl Environ Microbiol 69:4814–4822CrossRefGoogle Scholar
  21. Peng JS, Tsai WC, Chou CC (2002) Inactivation and removal of Bacillus cereus by sanitizer and detergent. Int J Food Microbiol 77:11–18CrossRefGoogle Scholar
  22. Raad I, Chatzinikolaou I, Chaiban G, Hanna H, Hachem R, Dvorak T, Cook G, Costerton W (2003) In vitro and ex vivo activities of minocycline and EDTA against microorganisms embedded in biofilm on catheter surfaces. Antimicrob Agents Chemother 47:3580–3585CrossRefGoogle Scholar
  23. Rasch M, Rasmussen TB, Andersen JB, Persson T, Nielsen J, Givskov M, Gram L (2007) Well-known quorum sensing inhibitors do not affect bacterial quorum sensing-regulated bean sprout spoilage. J Appl Microbiol 102:826–837CrossRefGoogle Scholar
  24. Rasmussen TB, Givskov M (2006) Quorum sensing inhibitors: a bargain of effects. Microbiology 152:895–904CrossRefGoogle Scholar
  25. Rasmussen TB, Skindersoe ME, Bjarnsholt T, Phipps RK, Christensen KB, Jensen PO, Andersen JB, Koch B, Larsen TO, Hentzer M, Eberl L, Hoiby N, Givskov M (2005a) Screening for quorum-sensing inhibitors (QSI) by use of a novel genetic system, the QSI selector. J Bacteriol 187:1799–1814CrossRefGoogle Scholar
  26. Rasmussen TB, Skindersoe ME, Bjarnsholt T, Phipps RK, Christensen KB, Jensen PO, Andersen JB, Koch B, Larsen TO, Hentzer M, Eberl L, Hoiby N, Givskov M (2005b) Identity and effects of quorum-sensing inhibitors produced by Penicillium species. Microbiology 151:1325–1340CrossRefGoogle Scholar
  27. Ren D, Sims JJ, Wood TK (2002) Inhibition of biofilm formation and swarming of Bacillus subtilis by (5Z)-4-bromo-5-(bromomethylene)-3-butyl-2(5H)-furanone. Lett Appl Microbiol 34:293–299CrossRefGoogle Scholar
  28. Russell AD (2003) Bacterial outer membrane and cell wall penetration and cell destruction by polluting chemical agents and physical conditions. Sci Prog 86:283–311CrossRefGoogle Scholar
  29. Stewart PS (2003) Diffusion in biofilms. J Bacteriol 185:1485–1491CrossRefGoogle Scholar
  30. Surette MG, Bassler BL (1998) Quorum sensing in Escherichia coli and Salmonella typhimurium. Proc Natl Acad Sci U S A 95:7046–7050CrossRefGoogle Scholar
  31. Surette MG, Miller MB, Bassler BL (1999) Quorum sensing in Escherichia coli, Salmonella typhimurium, and Vibrio harveyi: a new family of genes responsible for autoinducer production. Proc Natl Acad Sci U S A 96:1639–1644CrossRefGoogle Scholar
  32. Sutherland I (2001) Biofilm EPS: a strong and sticky framework. Microbiology 147:3–9Google Scholar
  33. Tarkkanen AM, Allen BL, Williams PH, Kauppi M, Haahtela K, Siitonen A, Orskov I, Orskov F, Clegg S, Korhonen TK (1992) Fimbriation, capsulation, and iron-scavenging systems of Klebsiella strains associated with human urinary tract infection. Infect Immun 60:1187–1192Google Scholar
  34. Teitzel GM, Parsek MR (2003) Heavy metal resistance of biofilm and planktonic Pseudomonas aeruginosa. Appl Environ Microbiol 69:2313–2320CrossRefGoogle Scholar
  35. Tuttlebee CM, O’Donnell MJ, Keane CT, Russell RJ, Sullivan DJ, Falkiner F, Coleman DC (2002) Effective control of dental chair unit waterline biofilm and marked reduction of bacterial contamination of output water using two peroxide-based disinfectants. J Hosp Infect 52:192–205CrossRefGoogle Scholar
  36. Vaara M (1992) Agents that increase the permeability of the outer membrane. Microbiol Rev 56:395–411Google Scholar
  37. Walker JT, Bradsha DJ, Fulford MR, Marsh PD (2003) Microbiological evaluation of a range of disinfectant products to control mixed-species biofilm contamination in a laboratory model of a dental unit water system. Appl Environ Microbiol 69:3327–3332CrossRefGoogle Scholar
  38. Wang L, Hashimoto Y, Tsao C-Y, Valdes JJ, Bentley WE (2005) cAMP and cAMP receptor protein (CRP) influence both synthesis and uptake of extracellular autoinducer-2 in Escherichia coli. J Bacteriol 187:2066–2076CrossRefGoogle Scholar
  39. Williams P (2002) Quorum sensing: an emerging target for antibacterial chemotherapy. Expert Opin Ther Targets 6:1–18CrossRefGoogle Scholar
  40. Xavier KB, Bassler BL (2003) LuxS quorum sensing: more than just a numbers game. Curr Opin Microbiol 6:191–197CrossRefGoogle Scholar
  41. Xavier KB, Bassler BL (2005) Regulation of uptake and processing of the quorum-sensing autoinducer AI-2 in Escherichia coli. J Bacteriol 187:238–248CrossRefGoogle Scholar
  42. Yakandawala N, Gawande PV, LoVetri K, Madhyastha S (2007) Effect of ovotransferrin, protamine sulfate and EDTA combination on biofilm formation by catheter-associated bacteria. J Appl Microbiol 102:722–727CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Iram Liaqat
    • 1
  • Robert Th. Bachmann
    • 2
    • 3
  • Anjum Nasim Sabri
    • 1
  • Robert G. J. Edyvean
    • 2
  • Catherine A. Biggs
    • 2
  1. 1.Department of Microbiology and Molecular GeneticsUniversity of the PunjabLahorePakistan
  2. 2.Department of Chemical and Process EngineeringThe University of SheffieldSheffieldUK
  3. 3.Malaysian Institute of Chemical and Bioengineering TechnologyUniversity Kuala LumpurAlor GajahMalaysia

Personalised recommendations