Phenotypic changes in pseudomonas aeruginosa induced by sub-inhibitory exposure to chlorohexidine

  • Sayran Hamad Haji Department of pharmacogonsy, College of Pharmacy, Hawler Medical University, Erbil, Iraq.
  • Safaa Toma Aka Department of pharmacogonsy, College of Pharmacy, Hawler Medical University, Erbil, Iraq.
Keywords: Biofilm, Chlorhexidine, Antibiotics, P. aeruginosa

Abstract

Background and objective: Many bacterial isolates show an increased antimicrobial resistance due to biofilm production. Repeated exposure to sub lethal concentrations of antimicrobial agents certainly contributes to the resistance as compared to planktonic bacteria. The aim of this study was to test whether the bacterial phenotypes of P. aeruginosa can be changed during exposures to the concentrations at sub lethal doses of chlorhexidine.

Methods: Sheep blood agar plates were used for evaluation of haemolysin assay for isolates of P. aeroginusa. A 96-flat bottom well microtiter plates were used for determination of MIC of antibiotic and biofilm formation. 

Results: All tested isolates were able to lyse RBCs after exposure to sub-MIC of chlorhexidine. Effectiveness of sub-lethal doses of chlorhexidine on biofilm formations varied depending on the contact time. In general, long contact time exhibited increasing biofilm than short time. No significant difference in biofilm was detected among contact times: day I, day II and day III (P = 0.132, P = 0.139 and P = 0.125, respectively). The most effective sub-MIC of CHX was against azithromycin, since the resistance increased significantly (P = 0.008).

Conclusion: Surviving P. aeruginosa to low concentration of chlorhexidine can exhibit stronger biofilm and increased resistance to antibiotics.

References

Houari A, Di Martino P. Effect of chlorhexidine and benzalkonium chloride on bacterial biofilm formation. Lett Appl Microbiol 2007; 45:652-6.

Chawner JA, Gilbert P. A comparative study of the bactericidal and growth inhibitory activities of the bisbiguanides alexidine and chlorhexidine. J Appl Bacteriol 1989; 66:243-52.

Pantanella F. Valenti P. Natalizi T. Passeri D. Analytical technique to study microbial biofilm on a biotic surface: pros and cons of main techniques currently in use. Ann Ig 2013; 25(1):31-42.

Russell AD. Antibiotic and biocide resistance in bacteria: Introduction. J Appl Microbiol Symposium 2002; Supp 92:1S-3S.

Masadeh MM, Mhaidat NM, Alzoubi KH, Hussein EI, Al-Trad EI. In vitro determination of the antibiotic susceptibility of biofilm-forming Pseudomonas aeruginosa and Staphylococcus aureus: possible role of proteolytic activity and membrane lipopolysaccharide. Infect Drug Resist 2013; 6:27-32.

Braoudaki M, Hilton AC. Adaptative resistance to biocides in Salmonella enterica and Escherichia coli O157 and cross resistance to antimicrobial agents. J Clin Microbiol 2004; 42(1):73-8.

Mazzola PG, Jozala AF, Novaes LC, Moriel P, Penna TC. Minimal inhibitory concentration (MIC) determination of disinfectant and/or sterilizing agents. Brazil J Pharm Sci 2009; 45(2):241-8.

Davis J, Spiegelman GB, Yim G. The world of sub-inhibitory antibiotic concentrations. Curr Opin Microbiol 2006; 9(5):445-53.

Kaplan JB. Antibiotic induced biofilm formation. Int J Artif Organs 2011; 34(9):737-51.

Mdonnell G, Russell AD. Antiseptics and Disinfectants: Activity, Action, and Resistance. Clin Microbiol Rev 1999; 12(1):147-79.

Augustin M. Ali-Vehmas T. Assessment of enzymatic cleaning agents and disinfectants against bacterial biofilms. Pharm Pharmaceut Sci 2004; 7(1): 55-64.

Fazlara A, Ekhtelat M. The Disinfectant Effects of Benzalkonium Chloride on Some Important Foodborne Pathogens. American-Eurasian J Agric Environ Sci 2012; 12(1):23-9.

Dynes JJ, Lawrence JR, Korber DR, Swerhone GD, Hitchcock AP. Morphlogical and biochemical changes in Pseudomonas florescens biofilms induced by sub-inhibitory Exposure to antimicrobial agents. Can J Microbiol 2009; 55:163-78.

Bisen PS, Debnath M, Prasad GB, Microbes concepts and applications. Newjersy: John Wiley and sons Inc; 2012.

Anton NN. Bacteriological study of discharging ear and evaluation of the effect of certain antibacterial agents against biofilm forming bacteria. MSc Thesis in Microbiology. Hawler Medical University, College of Medicine; 2013.

Griffiths BB, McClain O. The role of iron in the growth and hemolysin (streptolysin S) production in Streptococcus pyogenes. J Basic Microbiol 2007; 28(7):427-36.

Russell AD. Mechanisms of bacterial resistance to biocides. Int Biodeterior Biodegrad 1995; 36:247-65.

Prince HN, Nonmarker WS, Norgard RC, Prince DL. Drug resistance with topical antiseptics. J Pharm Sci 1978; 67:1629-31.

McDonnell G, Russell D. Antiseptics and disinfectants: Activity, Action and Resistance. Clin Microbiol Rev1999; 12(1):147-227.

Hugo WB, Pallent IJ, Grant DJ, Denyer SP, Davis A. Factors contributing to the survival of strain of Pseudomonas sepcia in chlorhexidine solution. Lett Appl Microbiol 1986; 2:37-42.

Russell AD. Mechanism of bacterial resistance to non-antibiotics: food additive and pharmaceutical preservatives. J Appl Bacteriol 1990; 71:191-201.

Thomas L, Russell AD, Maillard JY. Antimicrobial activity of chlorhexidine diastate and benzalkonium chloride against Pseudomonas aeruginosa and its response to biocide residues. J Appl Microbiol 2005; 98:533-43.

Andrel JF, Franklin MJ, Stewart PS. Role of antibiotic penetration limitation in Klebsiella pneumoniae biofilm resistance to ampicillin and ciprofloxacin. Antimicrob Agents Chemother 2000; 44:1818-24.

Xavier M, Macia D, Mena A, Juan C, Perez J, Oliver A. Azithromycin in Pseudomonas aeruginosa Biofilms: Bactericidal Activity and Selection of nfxB Mutants. Antimicrob Agents Chemother 2009; 53(4):1552-60.

Corona F, Martinez J. Phenotypic resistance to antibiotic. Antibiotic 2013; 2(2):237-55.

Published
2018-10-03
How to Cite
Haji, S., & Aka, S. (2018). Phenotypic changes in pseudomonas aeruginosa induced by sub-inhibitory exposure to chlorohexidine. Zanco Journal of Medical Sciences (Zanco J Med Sci), 18(3), 844-852. https://doi.org/10.15218/zjms.2014.0044
Section
Original Articles