Quorum sensing-regulated functions of Serratia marcescens are reduced by eugenol
Abstract
Background and Objectives: Serratia marcescens has emerged as a nosocomial pathogen responsible for human infections, where antibiotic resistance further complicates the treatments. In S. marcescens, biofilm formation and virulence factor production are controlled via quorum sensing (QS) system. QS is a signaling system that enables gene regulation to control diverse physiological functions in bacteria. Essential oils have shown to be potential in diminishing the pathogenicity and virulence of drug-resistant bacteria. This study was performed to determine whether eugenol would affect QS system, biofilm formation and virulence factor production of S. marcescens.
Materials and Methods: Biofilm formation, extracellular virulence factor production (hemolysin and protease), swarming motility and pigment formation of S. marcescens ATCC 13880 and S. marcescens Sm2 were assessed after eugenol exposure at 1.25 and 2.5 µg/ml concentrations. The expression of genes involved in motility (flhD), attachment (fimC), biofilm formation (bsmB, bsmA), and QS regulatory (swrR) were also evaluated.
Results: Eugenol treatment at 1.25 and 2.5 µg/ml concentrations caused a significant reduction in biofilm formation. The pigment, hemolysin and protease production of two studied S. marcescens strains, also reduced significantly by eugenol treatments (p<0.05). The bsmA, bsmB, flhD and fimC genes were down-regulated after eugenol treatment. The swrR gene expression was also reduced significantly by eugenol in both S. marcescens strains (p<0.05).
Conclusion: Eugenol inhibited quorum sensing-regulated functions of two studied S. marcescens strains.
1. Kida Y, Inoue H, Shimizu T, Kuwano K. Serratia marcescens serralysin induces inflammatory responses through protease-activated receptor 2. Infect Immun 2007;75:164-174.
2. Hejazi A, Falkiner F. Serratia marcescens. J Med Microbiol 1997;46:903-912.
3. Hawver LA, Jung SA, Ng W-L. Specificity and complexity in bacterial quorum-sensing systems. FEMS Microbiol Rev 2016;40:738-752.
4. Eberl L, Christiansen G, Molin S, Givskov M. Differentiation of Serratia liquefaciens into swarm cells is controlled by the expression of the flhD master operon. J Bacteriol 1996;178:554-559.
5. Coulthurst SJ, Williamson NR, Harris AK, Spring DR, Salmond GP. Metabolic and regulatory engineering of Serratia marcescens: mimicking phage-mediated horizontal acquisition of antibiotic biosynthesis and quorum-sensing capacities. Microbiology (Reading) 2006;152:1899-1911.
6. Rice S, Koh K, Queck S, Labbate M, Lam K, Kjelleberg S. Biofilm formation and sloughing in Serratia marcescens are controlled by quorum sensing and nutrient cues. J Bacteriol 2005;187:3477-3485.
7. Van Houdt R, Givskov M, Michiels CW. Quorum sensing in Serratia. FEMS Microbiol Rev 2007;31:407-424.
8. Morohoshi T, Shiono T, Takidouchi K, Kato M, Kato N, Kato J, et al. Inhibition of quorum sensing in Serratia marcescens AS-1 by synthetic analogs of N-acylhomoserine lactone. Appl Environ Microbiol 2007;73:6339-6344.
9. Brackman G, Coenye T. Quorum sensing inhibitors as anti-biofilm agents. Curr Pharm Des 2015;21:5-11.
10. Rasmussen TB, Givskov M. Quorum-sensing inhibitors as anti-pathogenic drugs. Int J Med Microbiol 2006;296(2-3):149-161.
11. Khan MSA, Zahin M, Hasan S, Husain FM, Ahmad I. Inhibition of quorum sensing regulated bacterial functions by plant essential oils with special reference to clove oil. Lett Appl Microbiol 2009;49:354-360.
12. Reichling J, Schnitzler P, Suschke U, Saller R. Essential oils of aromatic plants with antibacterial, antifungal, antiviral, and cytotoxic properties–an overview. Forsch Komplementmed 2009;16:79-90.
13. Kalemba D, Kunicka A. Antibacterial and antifungal properties of essential oils. Curr Med Chem 2003;10:813-829.
14. Bakkali F, Averbeck S, Averbeck D, Idaomar M. Biological effects of essential oils–a review. Food Chem Toxicol 2008;46:446-475.
15. Vattem DA, Mihalik K, Crixell SH, McLean RJ. Dietary phytochemicals as quorum sensing inhibitors. Fitoterapia 2007;78:302-310.
16. Walsh SE, Maillard JY, Russell A, Catrenich C, Charbonneau D, Bartolo R. Activity and mechanisms of action of selected biocidal agents on Gram-positive and-negative bacteria. J Appl Microbiol 2003;94:240-247.
17. Zhou L, Zheng H, Tang Y, Yu W, Gong Q. Eugenol inhibits quorum sensing at sub-inhibitory concentrations. Biotechnol Lett 2013;35:631-637.
18. Rathinam P, Vijay Kumar H, Viswanathan P. Eugenol exhibits anti-virulence properties by competitively binding to quorum sensing receptors. Biofouling 2017;33:624-639.
19. Wikler MA. “Performance Standards for Antimicrobial Susceptibility Testing, Sixteenth Informational Supplement, M100-S16,” Clinical and Laboratory Standards Institute (CLSI), Vol. 26, No. 3, 2006, Pennsylvania.
20. Adonizio A, Kong K-F, Mathee K. Inhibition of quorum sensing-controlled virulence factor production in Pseudomonas aeruginosa by South Florida plant extracts. Antimicrob Agents Chemother 2008;52:198-203.
21. Kashef N, Akbarizare M, Kamrava SK. Effect of sub-lethal photodynamic inactivation on the antibiotic susceptibility and biofilm formation of clinical Staphylococcus aureus isolates. Photodiagnosis Photodyn Ther 2013;10:368-373.
22. Gowrishankar S, Poornima B, Pandian SK. Inhibitory efficacy of cyclo (l-leucyl-l-prolyl) from mangrove rhizosphere bacterium–Bacillus amyloliquefaciens (MMS-50) toward cariogenic properties of Streptococcus mutans. Res Microbiol 2014;165:278-289.
23. Sethupathy S, Shanmuganathan B, Kasi PD, Pandian SK. Alpha-bisabolol from brown macroalga Padina gymnospora mitigates biofilm formation and quorum sensing controlled virulence factor production in Serratia marcescens. J Appl Phycol 2016;28:1987-1996.
24. Bakkiyaraj D, Sivasankar C, Pandian SK. Inhibition of quorum sensing regulated biofilm formation in Serratia marcescens causing nosocomial infections. Bioorg Med Chem Lett 2012;22:3089-3094.
25. Devi KR, Srinivasan S, Ravi AV. Inhibition of quorum sensing-mediated virulence in Serratia marcescens by Bacillus subtilis R-18. Microb Pathog 2018;120:166-175.
26. Salini R, Pandian SK. Interference of quorum sensing in urinary pathogen Serratia marcescens by Anethum graveolens. Pathog Dis 2015;73:ftv038.
27. Dubois M, Gilles KA, Hamilton JK, Rebers Pt, Smith F. Colorimetric method for determination of sugars and related substances. Anal Chem 1956;28:350-356.
28. Fekrirad Z, Kashef N, Arefian E. Photodynamic inactivation diminishes quorum sensing-mediated virulence factor production and biofilm formation of Serratia marcescens. World J Microbiol Biotechnol 2019;35:191.
29. Jakobsen TH, Bragason SK, Phipps RK, Christensen LD, van Gennip M, Alhede M, et al. Food as a source for quorum sensing inhibitors: iberin from horseradish revealed as a quorum sensing inhibitor of Pseudomonas aeruginosa. Appl Environ Microbiol 2012;78:2410-2421.
30. Taganna JC, Quanico JP, Perono RMG, Amor EC, Rivera WL. Tannin-rich fraction from Terminalia catappa inhibits quorum sensing (QS) in Chromobacterium violaceum and the QS-controlled biofilm maturation and LasA staphylolytic activity in Pseudomonas aeruginosa. J Ethnopharmacol 2011;134:865-871.
31. Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections.
Science 1999;284:1318-1322.
32. De Kievit TR, Gillis R, Marx S, Brown C, Iglewski BH. Quorum-sensing genes in Pseudomonas aeruginosa biofilms: their role and expression patterns. Appl Environ Microbiol 2001;67:1865-1873.
33. Yildiz FH, Schoolnik GK. Vibrio cholerae O1 El Tor: identification of a gene cluster required for the rugose colony type, exopolysaccharide production, chlorine resistance, and biofilm formation. Proc Natl Acad Sci U S A 1999;96:4028-4033.
34. Shanks RM, Lahr RM, Stella NA, Arena KE, Brothers KM, Kwak DH, et al. A Serratia marcescens PigP homolog controls prodigiosin biosynthesis, swarming motility and hemolysis and is regulated by cAMP-CRP and HexS. PLoS One 2013;8(3):e57634.
35. Packiavathy IASV, Priya S, Pandian SK, Ravi AV. Inhibition of biofilm development of uropathogens by curcumin–an anti-quorum sensing agent from Curcuma longa. Food Chem 2014;148:453-460.
36. Rosenberg M, Blumberger Y, Judes H, Bar-Ness R, Rubinstein E, Mazor Y. Cell surface hydrophobicity of pigmented and nonpigmented clinical Serratia marcescens strains. Infect Immun 1986;51:932-935.
37. Schiebel E, Braun V. Integration of the Serratia marcescens haemolysin into human erythrocyte membranes. Mol Microbiol 1989;3:445-453.
38. König W, Faltin Y, Scheffer J, Schöffler H, Braun V. Role of cell-bound hemolysin as a pathogenicity factor for Serratia infections. Infect Immun 1987;55:2554-2561.
39. Hertle R, Hilger M, Weingardt-Kocher S, Walev I. Cytotoxic action of Serratia marcescens hemolysin on human epithelial cells. Infect Immun 1999;67:817-825.
40. Srinivasan R, Mohankumar R, Kannappan A, Karthick Raja V, Archunan G, Karutha Pandian S, et al. Exploring the anti-quorum sensing and antibiofilm efficacy of phytol against Serratia marcescens associated acute pyelonephritis infection in Wistar rats. Front Cell Infect Microbiol 2017;7:498.
41. Labbate M, Zhu H, Thung L, Bandara R, Larsen MR, Willcox MD, et al. Quorum-sensing regulation of adhesion in Serratia marcescens MG1 is surface dependent. J Bacteriol 2007;189:2702-2711.
42. Liu J-H, Lai M-J, Ang S, Shu J-C, Soo P-C, Horng Y-T, et al. Role of flhDC in the expression of the nuclease gene nucA, cell division and flagellar synthesis in Serratia marcescens. J Biomed Sci 2000;7:475-483.
43. Srinivasan R, Devi KR, Kannappan A, Pandian SK, Ravi AV. Piper betle and its bioactive metabolite phytol mitigates quorum sensing mediated virulence factors and biofilm of nosocomial pathogen Serratia marcescens in vitro. J Ethnopharmacol 2016;193:592-603.
44. Mahlen SD. Serratia infections: from military experiments to current practice. Clin Microbiol Rev 2011;24(4):755-791.
45. Eberl L, Molin S, Givskov M. Surface motility of Serratia liquefaciens MG1. J Bacteriol 1999;181:1703-1712.
2. Hejazi A, Falkiner F. Serratia marcescens. J Med Microbiol 1997;46:903-912.
3. Hawver LA, Jung SA, Ng W-L. Specificity and complexity in bacterial quorum-sensing systems. FEMS Microbiol Rev 2016;40:738-752.
4. Eberl L, Christiansen G, Molin S, Givskov M. Differentiation of Serratia liquefaciens into swarm cells is controlled by the expression of the flhD master operon. J Bacteriol 1996;178:554-559.
5. Coulthurst SJ, Williamson NR, Harris AK, Spring DR, Salmond GP. Metabolic and regulatory engineering of Serratia marcescens: mimicking phage-mediated horizontal acquisition of antibiotic biosynthesis and quorum-sensing capacities. Microbiology (Reading) 2006;152:1899-1911.
6. Rice S, Koh K, Queck S, Labbate M, Lam K, Kjelleberg S. Biofilm formation and sloughing in Serratia marcescens are controlled by quorum sensing and nutrient cues. J Bacteriol 2005;187:3477-3485.
7. Van Houdt R, Givskov M, Michiels CW. Quorum sensing in Serratia. FEMS Microbiol Rev 2007;31:407-424.
8. Morohoshi T, Shiono T, Takidouchi K, Kato M, Kato N, Kato J, et al. Inhibition of quorum sensing in Serratia marcescens AS-1 by synthetic analogs of N-acylhomoserine lactone. Appl Environ Microbiol 2007;73:6339-6344.
9. Brackman G, Coenye T. Quorum sensing inhibitors as anti-biofilm agents. Curr Pharm Des 2015;21:5-11.
10. Rasmussen TB, Givskov M. Quorum-sensing inhibitors as anti-pathogenic drugs. Int J Med Microbiol 2006;296(2-3):149-161.
11. Khan MSA, Zahin M, Hasan S, Husain FM, Ahmad I. Inhibition of quorum sensing regulated bacterial functions by plant essential oils with special reference to clove oil. Lett Appl Microbiol 2009;49:354-360.
12. Reichling J, Schnitzler P, Suschke U, Saller R. Essential oils of aromatic plants with antibacterial, antifungal, antiviral, and cytotoxic properties–an overview. Forsch Komplementmed 2009;16:79-90.
13. Kalemba D, Kunicka A. Antibacterial and antifungal properties of essential oils. Curr Med Chem 2003;10:813-829.
14. Bakkali F, Averbeck S, Averbeck D, Idaomar M. Biological effects of essential oils–a review. Food Chem Toxicol 2008;46:446-475.
15. Vattem DA, Mihalik K, Crixell SH, McLean RJ. Dietary phytochemicals as quorum sensing inhibitors. Fitoterapia 2007;78:302-310.
16. Walsh SE, Maillard JY, Russell A, Catrenich C, Charbonneau D, Bartolo R. Activity and mechanisms of action of selected biocidal agents on Gram-positive and-negative bacteria. J Appl Microbiol 2003;94:240-247.
17. Zhou L, Zheng H, Tang Y, Yu W, Gong Q. Eugenol inhibits quorum sensing at sub-inhibitory concentrations. Biotechnol Lett 2013;35:631-637.
18. Rathinam P, Vijay Kumar H, Viswanathan P. Eugenol exhibits anti-virulence properties by competitively binding to quorum sensing receptors. Biofouling 2017;33:624-639.
19. Wikler MA. “Performance Standards for Antimicrobial Susceptibility Testing, Sixteenth Informational Supplement, M100-S16,” Clinical and Laboratory Standards Institute (CLSI), Vol. 26, No. 3, 2006, Pennsylvania.
20. Adonizio A, Kong K-F, Mathee K. Inhibition of quorum sensing-controlled virulence factor production in Pseudomonas aeruginosa by South Florida plant extracts. Antimicrob Agents Chemother 2008;52:198-203.
21. Kashef N, Akbarizare M, Kamrava SK. Effect of sub-lethal photodynamic inactivation on the antibiotic susceptibility and biofilm formation of clinical Staphylococcus aureus isolates. Photodiagnosis Photodyn Ther 2013;10:368-373.
22. Gowrishankar S, Poornima B, Pandian SK. Inhibitory efficacy of cyclo (l-leucyl-l-prolyl) from mangrove rhizosphere bacterium–Bacillus amyloliquefaciens (MMS-50) toward cariogenic properties of Streptococcus mutans. Res Microbiol 2014;165:278-289.
23. Sethupathy S, Shanmuganathan B, Kasi PD, Pandian SK. Alpha-bisabolol from brown macroalga Padina gymnospora mitigates biofilm formation and quorum sensing controlled virulence factor production in Serratia marcescens. J Appl Phycol 2016;28:1987-1996.
24. Bakkiyaraj D, Sivasankar C, Pandian SK. Inhibition of quorum sensing regulated biofilm formation in Serratia marcescens causing nosocomial infections. Bioorg Med Chem Lett 2012;22:3089-3094.
25. Devi KR, Srinivasan S, Ravi AV. Inhibition of quorum sensing-mediated virulence in Serratia marcescens by Bacillus subtilis R-18. Microb Pathog 2018;120:166-175.
26. Salini R, Pandian SK. Interference of quorum sensing in urinary pathogen Serratia marcescens by Anethum graveolens. Pathog Dis 2015;73:ftv038.
27. Dubois M, Gilles KA, Hamilton JK, Rebers Pt, Smith F. Colorimetric method for determination of sugars and related substances. Anal Chem 1956;28:350-356.
28. Fekrirad Z, Kashef N, Arefian E. Photodynamic inactivation diminishes quorum sensing-mediated virulence factor production and biofilm formation of Serratia marcescens. World J Microbiol Biotechnol 2019;35:191.
29. Jakobsen TH, Bragason SK, Phipps RK, Christensen LD, van Gennip M, Alhede M, et al. Food as a source for quorum sensing inhibitors: iberin from horseradish revealed as a quorum sensing inhibitor of Pseudomonas aeruginosa. Appl Environ Microbiol 2012;78:2410-2421.
30. Taganna JC, Quanico JP, Perono RMG, Amor EC, Rivera WL. Tannin-rich fraction from Terminalia catappa inhibits quorum sensing (QS) in Chromobacterium violaceum and the QS-controlled biofilm maturation and LasA staphylolytic activity in Pseudomonas aeruginosa. J Ethnopharmacol 2011;134:865-871.
31. Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections.
Science 1999;284:1318-1322.
32. De Kievit TR, Gillis R, Marx S, Brown C, Iglewski BH. Quorum-sensing genes in Pseudomonas aeruginosa biofilms: their role and expression patterns. Appl Environ Microbiol 2001;67:1865-1873.
33. Yildiz FH, Schoolnik GK. Vibrio cholerae O1 El Tor: identification of a gene cluster required for the rugose colony type, exopolysaccharide production, chlorine resistance, and biofilm formation. Proc Natl Acad Sci U S A 1999;96:4028-4033.
34. Shanks RM, Lahr RM, Stella NA, Arena KE, Brothers KM, Kwak DH, et al. A Serratia marcescens PigP homolog controls prodigiosin biosynthesis, swarming motility and hemolysis and is regulated by cAMP-CRP and HexS. PLoS One 2013;8(3):e57634.
35. Packiavathy IASV, Priya S, Pandian SK, Ravi AV. Inhibition of biofilm development of uropathogens by curcumin–an anti-quorum sensing agent from Curcuma longa. Food Chem 2014;148:453-460.
36. Rosenberg M, Blumberger Y, Judes H, Bar-Ness R, Rubinstein E, Mazor Y. Cell surface hydrophobicity of pigmented and nonpigmented clinical Serratia marcescens strains. Infect Immun 1986;51:932-935.
37. Schiebel E, Braun V. Integration of the Serratia marcescens haemolysin into human erythrocyte membranes. Mol Microbiol 1989;3:445-453.
38. König W, Faltin Y, Scheffer J, Schöffler H, Braun V. Role of cell-bound hemolysin as a pathogenicity factor for Serratia infections. Infect Immun 1987;55:2554-2561.
39. Hertle R, Hilger M, Weingardt-Kocher S, Walev I. Cytotoxic action of Serratia marcescens hemolysin on human epithelial cells. Infect Immun 1999;67:817-825.
40. Srinivasan R, Mohankumar R, Kannappan A, Karthick Raja V, Archunan G, Karutha Pandian S, et al. Exploring the anti-quorum sensing and antibiofilm efficacy of phytol against Serratia marcescens associated acute pyelonephritis infection in Wistar rats. Front Cell Infect Microbiol 2017;7:498.
41. Labbate M, Zhu H, Thung L, Bandara R, Larsen MR, Willcox MD, et al. Quorum-sensing regulation of adhesion in Serratia marcescens MG1 is surface dependent. J Bacteriol 2007;189:2702-2711.
42. Liu J-H, Lai M-J, Ang S, Shu J-C, Soo P-C, Horng Y-T, et al. Role of flhDC in the expression of the nuclease gene nucA, cell division and flagellar synthesis in Serratia marcescens. J Biomed Sci 2000;7:475-483.
43. Srinivasan R, Devi KR, Kannappan A, Pandian SK, Ravi AV. Piper betle and its bioactive metabolite phytol mitigates quorum sensing mediated virulence factors and biofilm of nosocomial pathogen Serratia marcescens in vitro. J Ethnopharmacol 2016;193:592-603.
44. Mahlen SD. Serratia infections: from military experiments to current practice. Clin Microbiol Rev 2011;24(4):755-791.
45. Eberl L, Molin S, Givskov M. Surface motility of Serratia liquefaciens MG1. J Bacteriol 1999;181:1703-1712.
| Files | ||
| Issue | Vol 12 No 5 (2020) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v12i5.4607 | |
| Keywords | ||
| Serratia marcescens; Virulence factors; Biofilm formation; Quorum sensing; Eugenol | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Fekrirad Z, Gattali B, Kashef N. Quorum sensing-regulated functions of Serratia marcescens are reduced by eugenol. Iran J Microbiol. 2020;12(5):451-459.
