The correlation between the presence of quorum sensing, toxin-antitoxin system genes and MIC values with ability of biofilm formation in clinical isolates of Pseudomonas aeruginosa
-
Saeed Hemati
,
Farid Azizi-Jalilian
,
Iraj Pakzad
,
Morovat Taherikalani
,
Abbas Maleki
,
Sajedeh Karimi
,
Azam Monjezei
,
Zahra Mahdavi
,
Mohamad Reza Fadavi
,
Kourosh Sayehmiri
,
Nourkhoda Sadeghifard
Abstract
Introduction: Pseudomonas aeruginosa is a Gram-negative bacterium that considered as important opportunistic human pathogen. One of the mechanisms that help bacteria to tolerate survival in adverse conditions and resistance to antibiotics is biofilm formation through quorum sensing (QS) signals and toxin-antitoxin (TA) systems. QS and TA are two systems that have important roles in biofilm formation. QS is a global regulatory mechanism that enable bacteria to communicate with each other by production of auto inducers (AI) molecules in population. Because of importance biofilm formation in P. aeruginosa infections, here, we studied frequency of QS and TA genes among clinical isolates of P. aeruginosa with ability of biofilm formation.
Materials and Methods: One hundred and forty clinical isolates of P. aeruginosa were collected from Tehran and Ilam hospitals. The isolates were identified by biochemical tests. Biofilm formation was evaluated by microplate method. After DNA extraction by boiling method, the frequency of QS genes (lasIR, rhlIR), and TA genes (mazEF, relBE, hipBA, ccdAB and mqsR) were analyzed by PCR.
Results: Our results showed that maximum resistance is related to aztreonam (72.85%) antibiotic. Most of isolates were able to produce biofilm (87.15%) and the majority of them formed strong biofilm (56.42%). PCR results showed that frequency of mazEF, relBE, hipBA, ccdAB, mqsR, lasIR and rhlIR genes were 85.71, 100, 1.42, 100, 57.14, 93.57 and 83.57 percent, respectively.
Conclusion: Clinical isolates of P. aeruginosa had high ability to form biofilm, and QS and TA system genes among these isolates were very high (except hipBA genes). There are significaut correlation between biofilm for mation and present of QS and TA system genes.
El Solh AA, Alhajhusain A. Update on the treatment of Pseudomonas aeruginosa pneumonia. J. Antimicrob Chemother 2009; dkp201.
Neidig A, Yeung AT, Rosay T, Tettmann B, Strempel N, Rueger M, et al. Typ A is involved in virulence, antimicrobial resistance and biofilm formation in Pseudomonas aeruginosa. BMC Microbiology 2013;
: 77.
Belshe RB, Edwards KM, Vesikari T, Black SV, Walker RE, Hultquist M, et al. Live attenuated versus inactivated influenza vaccine in infants and young children. . N Engl J Med 2007; 356: 685-696.
Klausen M, Heydorn A, Ragas P, Lambertsen L, Aaes- Jørgensen A, Molin S, et al. Biofilm formation by Pseudomonas aeruginosa wild type, flagella and type IV pili mutants. Mol Microbiol 2003; 48: 1511-1524.
Wang X, Wood TK. Toxin-antitoxin systems influence biofilm and persister cell formation and the general stress response. Appl Environ Microbiol 2011; 77:5577-5583.
O’Loughlin CT, Miller LC, Siryaporn A, Drescher K, Semmelhack MF, Bassler BL. A quorum-sensing inhibitor blocks Pseudomonas aeruginosa virulence and biofilm formation. PNAS 2013; 110: 17981-17986.
Hartmann A, Schikora A. Quorum sensing of bacteria and trans-kingdom interactions of N-acyl homoserine lactones with eukaryotes. J Chem Ecol 2012; 38: 704-713.
Szabó MÁ, Varga GZ, Hohmann J, Schelz Z, Szegedi E, Amaral L, et al. Inhibition of quorum-sensing signals by essential oils. Phytother Res 2010; 24: 782-786.
Mutschler H, Gebhardt M, Shoeman RL, Meinhart A. A novel mechanism of programmed cell death in bacteria by toxin–antitoxin systems corrupts peptidoglycan synthesis. PLoS Biology 2011; 9: e1001033.
Schuster CF, Bertram R. Toxin–antitoxin systems are ubiquitous and versatile modulators of prokaryotic cell fate. FEMS Microbiol Lett 2013; 340: 73-85.
Yamaguchi Y, Park J-H, Inouye M. Toxin-antitoxin systems in bacteria and archaea. Annu Rev Genet 2011; 45: 61-79.
Leplae R, Geeraerts D, Hallez R, Guglielmini J, Drèze P, Van Melderen L. Diversity of bacterial type II toxin–antitoxin systems: a comprehensive search and functional analysis of novel families. Nucleic Acids Res 2011: gkr131.
Park SJ, Son WS, Lee B-J. Structural overview of toxin– antitoxin systems in infectious bacteria: A target for developing antimicrobial agents. Biochim Biophys Acta 2013; 1834: 1155-1167.
Ghafourian S, Raftari M, Sadeghifard N, Sekawi Z.Toxin-antitoxin Systems: Classifcation, Biological Function and Application in Biotechnology. CIMB, Curr Iss Mol Biol 2013; 16: 9-14.
Percival SL, Hill KE, Malic S, Thomas DW, Williams DW. Antimicrobial tolerance and the significance of persister cells in recalcitrant chronic wound biofilms. Wound Repair Regen 2011; 19: 1-9.
Queipo-Ortuño MI, Colmenero JDD, Macias M, Bravo MJ, Morata P. Preparation of bacterial DNA template by boiling and effect of immunoglobulin G as an inhibitor in real-time PCR for serum samples from patients with brucellosis. Clin Vaccine Immunol 2008; 15: 293-296.
Yamaguchi Y, Inouye M. Regulation of growth and death in Escherichia coli by toxin–antitoxin systems. Nature Rev Microbiol 2011; 9: 779-790.
Yang L, Rybtke MT, Jakobsen TH, Hentzer M, Bjarnsholt T, Givskov M, et al. Computer-aided identification of recognized drugs as Pseudomonas aeruginosa quorum-sensing inhibitors. Antimicrob. Agents Chemother 2009; 53: 2432-43.
Crespo M, Woodford N, Sinclair A, Kaufmann M, Turton J, Glover J, et al. Outbreak of carbapenem- resistant Pseudomonas aeruginosa producing VIM-8, a novel metallo-β-lactamase, in a tertiary care center in Cali, Colombia. J CLIN MICROBIOL 2004; 42: 5094-101.
Mah T-FC OTG. Mechanisms of biofilm resistance to antimicrobial agents. TRENDS MICROBIOL 2001; 9: 34-9.
Williams JJ, Halvorsen EM, Dwyer EM, DiFazio RM, Hergenrother PJ. Toxin–antitoxin (TA) systems are prevalent and transcribed in clinical isolates of Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus. FEMS Microbiol Lett 2011;322: 41-50.
Baumann U, Mansouri E, Von Specht B-U. Recombinant OprF–OprI as a vaccine against< i> Pseudomonas aeruginosa infections. VACCINE 2004; 22: 840-7.23.Whiteley M, Bangera MG, Bumgarner RE, Parsek MR, Teitzel GM, Lory S, et al. Gene expression in Pseudomonas aeruginosa biofilms. NATURE 2001;413: 860-4.
Cabrol S, Olliver A, Pier GB, Andremont A, Ruimy R. Transcription of quorum-sensing system genes in clinical and environmental isolates of Pseudomonas aeruginosa. J BACTERIOL 2003; 185:7222-30.
Lemos JA, Brown TA, Abranches J, Burne RA.Characteristics of Streptococcus mutans strains lacking the MazEF and RelBE toxin–antitoxin modules. FEMS Microbiol Lett 2005; 253: 251-7.
Moritz EM, Hergenrother PJ. Toxin–antitoxin systems are ubiquitous and plasmid-encoded in vancomycin- resistant enterococci. Proc. Natl. Acad. Sci. U.S.A 2007; 104: 311-6.
Fauvart M, De Groote VN, Michiels J. Role of persister cells in chronic infections: clinical relevance and perspectives on anti-persister therapies. J MED MICROBIOL 2011; 60: 699-709.
Anderson G, O’toole G (2008). Bacterial biofilms. 1nd ed. Springer. Verlag Berlin Heidelberg.
Belitsky M, Avshalom H, Erental A, Yelin I, Kumar S, London N, et al. The Escherichia coli extracellular death factor EDF induces the endoribonucleolytic activities of the toxins MazF and ChpBK. Mol Cell 2011;4: 625-35.
Barrios AFG, Zuo R, Hashimoto Y, Yang L, Bentley WE, Wood TK. Autoinducer 2 controls biofilm formation in Escherichia coli through a novel motility quorum-sensing regulator (MqsR, B3022). J Bacteriol 2006; 188: 305-16.
Xu Z, Fang X, Wood TK, Huang ZJ. A systems-level approach for investigating Pseudomonas aeruginosa biofilm formation. PloS one 2013; 8: e57050.
Balaban N, Giacometti A, Cirioni O, Gov Y, Ghiselli R, Mocchegiani F, et al. Use of the quorum-sensing inhibitor RNAIII-inhibiting peptide to prevent biofilm formation in vivo by drug-resistant Staphylococcus epidermidis. J Infect Dis 2003; 187: 625-30.
Lioy VS, Rey O, Balsa D, Pellicer T, Alonso JC. A toxin–antitoxin module as a target for antimicrobial development. Plasmid 2010; 63: 31-9.
Williams JJ, Hergenrother PJ. Artifcial activation of toxin–antitoxin systems as an antibacterial strategy. Trends Microbiol 2012; 20: 291-8.
Kalia VC, Purohit HJ. Quenching the quorum sensing system: potential antibacterial drug targets. Crit Rev Microbiol 2011; 37: 121-40.
| Files | ||
| Issue | Vol 6 No 3 (2014) | |
| Section | Articles | |
| Keywords | ||
| P. aeruginosa Quorum sensing Toxin-antitoxin systems | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
