Susceptibility To Caspofungin And Fluconazole And Als1/Als3 Gene Expression In Biofilm And Dispersal Cells Of Candida Albicans / Profil Osjetljivosti Na Kaspofungin I Flukonazol I Ekspresija Gena Als1 I Als3 U Stanicama Biofilma Te Planktonskim Stanicama Vrste Candida Albicans

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Abstract

The biofilm of Candida albicans has been implicated as a source of bloodstream infections. Dispersal cells, as the final biofilm stage, are responsible for its spread. The aim of this study was to compare the susceptibility of biofilm and dispersal cells vs. planktonic cells (overnight liquid culture) of C. albicans to caspofungin (CAS) and fluconazole (FLU) when the drugs were added: i) at the beginning of the experiment; ii) after 1.5 h (adherence stage); iii) after 24 h (early mature biofilm). The findings were evaluated after 48 h (mature biofilm) using the XTT reduction assay. Later administration of the drug increased biofilm sessile minimal inhibitory concentration (SMIC80) of both FLU and CAS from 1 μg mL-1 to over 64 μg mL-1 and from 0.125 μg mL-1 to over 16 μg mL-1, respectively. Susceptibility of dispersal cells also decreased with time of administration.

We also determined the expression of the Als1 and Als3 genes in 48-h sessile biofilm and dispersal cells of C. albicans SC5314 and compared it to planktonic cells. The expression was normalised to the standard Act1 gene in every condition tested. Quantitative real-time PCR revealed a strong up-regulation of the Als1 gene in the dispersal cells but not in biofilm and high expression of the Als3 gene in both biofilm and dispersal cells. High expression of both Als1 and Als3 genes supports the hypothesis that dispersal cells pose a high-risk of infection.

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  • 1. Elias S Banin E. Multi-species biofilms: living with friendly neighbors. FEMS Microbiol Rev 2012;36:990-1004.

  • 2. Kojic EM Darouiche RO. Candida infections of medical devices. Clin Microbiol Rev 2004;17:255-67.

  • 3. Djeribi R Bouchloukh W Jouenne T Menaa B. Characterization of bacterial biofilms formed on urinary catheters. Am J Infect Control 2012;40:854-9.

  • 4. Hajjeh RA Sofair AN Harrison LH Lyon GM Arthington- Skaggs BA Mirza SA Phelan M Morgan J Lee-Yang W Ciblak MA Benjamin LE Sanza LT Huie S Yeo SF Brandt ME Warnock DW. Incidence of bloodstream infections due to Candida species and in vitro susceptibilities of isolates collected from 1998 to 2000 in a population-based active surveillance program. J Clin Microbiol 2004;42:1519-27.

  • 5. Mukherjee PK Chandra J. Candida biofilm resistance. Drug Resist Updat 2004;7:301-9.

  • 6. Borecka-Melkusova S Moran GP Sullivan DJ Kucharikova S Chorvat D Jr Bujdakova H. The expression of genes involved in the ergosterol biosynthesis pathway in Candidaalbicans and Candida dubliniensis biofilms exposed to fluconazole. Mycoses 2009;52:118-28.

  • 7. Walker LA Gow NA Munro CA. Fungal echinocandin resistance. Fungal Genet Biol 2010;47:117-26.

  • 8. Bal AM. The echinocandins: three useful choices or three too many? Int J Antimicrob Agents 2010;35:13-8.

  • 9. Kucharikova S Tournu H Holtappels M Van Dijck P Lagrou K. In vivo efficacy of anidulafungin against mature Candidaalbicans biofilms in a novel rat model of catheter-associated candidiasis. Antimicrob Agents Chemother 2010;54:4474-5.

  • 10. Fiori B Posteraro B Torelli R Tumbarello M Perlin DS Fadda G Sanguinetti M. In vitro activities of anidulafungin and other antifungal agents against biofilms formed by clinical isolates of different Candida and Aspergillus species. Antimicrob Agents Chemother 2011;55:3031-5.

  • 11. Kucharikova S Tournu H Lagrou K Van Dijck P Bujdakova H. Detailed comparison of Candida albicans and Candidaglabrata biofilms under different conditions and their susceptibility to caspofungin and anidulafungin. J Med Microbiol 2011;60:1261-9.

  • 12. Ramage G Vandewalle K Wickes BL Lopez-Ribot JL. Characteristics of biofilm formation by Candida albicans. Rev Iberoam Micol 2001;18:163-70.

  • 13. Finkel JS Mitchell AP. Genetic control of Candida albicans biofilm development. Nat Rev Microbiol 2011;9:109-18.

  • 14. Uppuluri P Chaturvedi AK Srinivasan A Banerjee M Ramasubramaniam AK Kohler JR Kadosh D Lopez-Ribot JL. Dispersion as an important step in the Candida albicans biofilm developmental cycle. PLoS Pathog 2010;26: e1000828.

  • 15. Uppuluri P Pierce CG Thomas DP Bubeck SS Saville SP Lopez-Ribot JL. The transcriptional regulator Nrg1p controls Candida albicans biofilm formation and dispersion. Eukaryot Cell 2010;9:1531-7.

  • 16. Nailis H Kucharikova S Řičicova M Van Dijck P Deforce D Nelis H Coenye T. Real-time PCR expression profiling of genes encoding potential virulence factors in Candidaalbicans biofilms: identification of model-dependent and - independent gene expression. BMC Microbiol 2010;16:114.

  • 17. Nett JE Lepak AJ Marchillo K Andes DR. Time course global gene expression analysis of an in vivo Candida biofilm. J Infect Dis 2009;200:307-13.

  • 18. Řičicova M Kucharikova S Tournu H Hendrix J Bujdakova H Van Eldere J Lagrou K Van Dijck P. Candida albicans biofilm formation in a new in vivo rat model. Microbiology 2010;156:909-19.

  • 19. Yeater KM Chandra J Cheng G Mukherjee PK Zhao X Rodriguez-Zas SL Kwast KE Ghannoum MA Hoyer LL. Temporal analysis of Candida albicans gene expression during biofilm development. Microbiology 2007;153:2373-85.

  • 20. Nailis H Vandenbroucke R Tilleman K Deforce D Nelis H Coenye T. Monitoring ALS1 and ALS3 gene expression during in vitro Candida albicans biofilm formation under continuous flow conditions. Mycopathologia 2009;167:9-17.

  • 21. Gillum AM Tsay EY Kirsch DR. Isolation of the Candidaalbicans gene for orotidine-5’-phosphate decarboxylase by complementation of S. cerevisiae ura3 and E. coli pyrF mutations. Mol Gen Genet 1984;198:179-82.

  • 22. National Committee for Clinical Laboratory Standards. Reference method for broth dilution antifungal susceptibility testing of yeast. Approved standard M27-A3. Wayne (PA): National Committee for Clinical Laboratory Standards; 2008.

  • 23. Li X Yan Z Xu J. Quantitative variation of biofilms among strains in natural populations of Candida albicans. Microbiolgy 2003;149:353-62.

  • 24. Livak KJ Schmittgen TD. Analysis of relative gene expression data using Real-Time quantitative PCR and the 2−ΔΔCT method. Methods 2001;25:402-8.

  • 25. Green CB Zhao X Yeater KM Hoyer LL. Construction and real-time RT-PCR validation of Candida albicans PALS-GFP reporter strains and their use in flow cytometry analysis of ALS gene expression in budding and filamenting cells. Microbiology 2003;151:1051-60.

  • 26. Ramage G Martinez JP Lopez-Ribot JL. Candida biofilms on implanted biomaterials: a clinically significant problem. FEMS Yeast Res 2006;6:979-86.

  • 27. Clancy CJ Yu VL Morris AJ Snydman DR Nguyen MH. Fluconazole MIC and the fluconazole dose/MIC ratio correlate with therapeutic response among patients with candidemia. Antimicrob Agents Chemother 2005;49:3171-7.

  • 28. Badiee P Alborzi A Davarpanah MA Shakiba E. Distributions and antifungal susceptibility of Candida species from mucosal sites in HIV positive patients. Arch Iran Med 2010;13:282-7.

  • 29. Martins HP da Silva MC Paiva LC Svidzinski TI Consolaro ME. Efficacy of fluconazole and nystatin in the treatment of vaginal Candida species. Acta Derm Venereol 2012;92:78-2.

  • 30. Pfaller MA Messer SA Moet GJ Jones RN Castanheira M. Candida bloodstream infections: comparison of species distribution and resistance to echinocandin and azole antifungal agents in Intensive Care Unit (ICU) and non-ICU settings in the SENTRY Antimicrobial Surveillance Program (2008-2009). Int J Antimicrob Agents 2011;38:65-9.

  • 31. Uppuluri P Srinivasan A Ramasubramanian A Lopez-Ribot JL. Effects of fluconazole amphotericin B and caspofungin on Candida albicans biofilms under conditions of flow and on biofilm dispersion. Antimicrob Agents Chemother 2011;55:3591-3.

  • 32. Robbins N Uppuluri P Nett J Rajendran R Ramage G Lopez-Ribot JL Andes D Cowen LE. Hsp90 governs dispersion and drug resistance of fungal biofilms. PLoS Pathog 2011;7:e1002257.

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