1887

Abstract

species can form biofilms on tissues and medical devices, making them less susceptible to antifungal agents.

Antifungal combination may be an effective strategy to fight against biofilm.

In this study, we investigated the activity of fluconazole, caspofungin and amphotericin B, alone and in combination, against 17 clinical and 6 isolates with high biofilm formation. We also tested LL-37 and lysozyme for anti-biofilm activity against a selected isolate.

biofilms were prepared using the 96-well plate-based method. The minimum biofilm eradication concentrations were determined for single and combined antifungal drugs. The activity of LL-37 and lysozyme was determined by visual reading for planktonic cells and using the XTT assay for biofilms.

Under biofilm conditions, fluconazole plus caspofungin showed synergistic effects against 60.9% (14 of 23) of the tested isolates, including 70.6% of [fractional inhibitory concentration index (FICI), 0.26–1.03] and 33.3% of (FICI, 0.04–2.03) isolates. Using this combination, no antagonism was observed. Amphotericin B plus caspofungin showed no effects against 78.3% (18 of 23) of the tested isolates. Amphotericin B plus fluconazole showed no effects against 65.2% (15 of 23) of the tested isolates and may have led to antagonism against 2 . and 2 isolates. LL-37 and lysozyme had no effect on biofilms of the selected isolate.

We found that fluconazole plus caspofungin led to a synergistic effect against and biofilms. The efficacy of the antifungal combination therapies of the proposed schemes against biofilm-associated infections requires careful and constant evaluation.

Funding
This study was supported by the:
  • Chen-HsiangLee , Chang Gung Memorial Hospital , (Award CMRPG 8H0491)
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/content/journal/jmm/10.1099/jmm.0.001283
2020-11-30
2021-01-16
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References

  1. Kullberg BJ, Arendrup MC. Invasive candidiasis. N Engl J Med 2015; 373:1445–1456 [CrossRef][PubMed]
    [Google Scholar]
  2. Donlan RM, Costerton JW. Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 2002; 15:167–193 [CrossRef][PubMed]
    [Google Scholar]
  3. Taff HT, Mitchell KF, Edward JA, Andes DR. Mechanisms of Candida biofilm drug resistance. Future Microbiol 2013; 8:1325–1337 [CrossRef][PubMed]
    [Google Scholar]
  4. Ramage G, Rajendran R, Sherry L, Williams C. Fungal biofilm resistance. Int J Microbiol 2012; 2012:52852114 [CrossRef][PubMed]
    [Google Scholar]
  5. Tumbarello M, Posteraro B, Trecarichi EM, Fiori B, Rossi M et al. Biofilm production by Candida species and inadequate antifungal therapy as predictors of mortality for patients with candidemia. J Clin Microbiol 2007; 45:1843–1850 [CrossRef][PubMed]
    [Google Scholar]
  6. Tumbarello M, Fiori B, Trecarichi EM, Posteraro P, Losito AR et al. Risk factors and outcomes of candidemia caused by biofilm-forming isolates in a tertiary care hospital. PLoS One 2012; 7:e33705 [CrossRef][PubMed]
    [Google Scholar]
  7. Cavalheiro M, Teixeira MC. Threats Candida biofilms: challenges, and promising strategies. Front Med 2018; 5:28
    [Google Scholar]
  8. Pappas PG, Kauffman CA, Andes DR, Clancy CJ, Marr KA et al. Clinical practice guideline for the management of candidiasis: 2016 update by the infectious diseases Society of America. Clin Infect Dis 2016; 62:e1–e50 [CrossRef][PubMed]
    [Google Scholar]
  9. Kung HC, Huang PY, Chen WT, BS K, Chen YC et al. Guidelines for the use of antifungal agents in patients with invasive fungal diseases in Taiwan. J Microbiol Immunol Infect 2016; 2018:1–17
    [Google Scholar]
  10. Nucci M, Anaissie E, Betts RF, Dupont BF, Wu C et al. Early removal of central venous catheter in patients with candidemia does not improve outcome: analysis of 842 patients from 2 randomized clinical trials. Clin Infect Dis 2010; 51:295–303 [CrossRef][PubMed]
    [Google Scholar]
  11. Cui J, Ren B, Tong Y, Dai H, Zhang L. Synergistic combinations of antifungals and anti-virulence agents to fight against Candida albicans. Virulence 2015; 6:362–371 [CrossRef][PubMed]
    [Google Scholar]
  12. Tobudic S, Kratzer C, Lassnigg A, Graninger W, Presterl E. In vitro activity of antifungal combinations against Candida albicans biofilms. J Antimicrob Chemother 2010; 65:271–274 [CrossRef][PubMed]
    [Google Scholar]
  13. Touil HFZ, Boucherit-Otmani Z, Boucherit K. In vitro activity of antifungal combinations against planktonic and sessile cells of Candida albicans isolated from medical devices in an intensive care department. J Mycol Med 2018; 28:414–418 [CrossRef][PubMed]
    [Google Scholar]
  14. Choi MJ, Won EJ, Shin JH, Kim SH, Lee W-G et al. Resistance Mechanisms and Clinical Features of Fluconazole-Nonsusceptible Candida tropicalis Isolates Compared with Fluconazole-Less-Susceptible Isolates. Antimicrob Agents Chemother 2016; 60:3653–3661 [CrossRef][PubMed]
    [Google Scholar]
  15. Melo AS, Bizerra FC, Freymüller E, Arthington-Skaggs BA, Colombo AL. Biofilm production and evaluation of antifungal susceptibility amongst clinical Candida spp. isolates, including strains of the Candida parapsilosis complex. Med Mycol 2011; 49:253–262 [CrossRef][PubMed]
    [Google Scholar]
  16. Li W-S, Chen Y-C, Kuo S-F, Chen F-J, Lee C-H. The impact of biofilm formation on the persistence of candidemia. Front Microbiol 2018; 9:1196 [CrossRef][PubMed]
    [Google Scholar]
  17. Černáková L, Light C, Salehi B, Rogel-Castillo C, Victoriano M et al. Novel therapies for biofilm-based Candida spp. infections. Adv Exp Med Biol 2019; 1214:93–123 [CrossRef][PubMed]
    [Google Scholar]
  18. Pulcrano G, Panellis D, De Domenico G, Rossano F, Catania MR. Ambroxol influences voriconazole resistance of Candida parapsilosis biofilm. FEMS Yeast Res 2012; 12:430–438 [CrossRef][PubMed]
    [Google Scholar]
  19. Fox EP, Singh-Babak SD, Hartooni N, Nobile CJ. Biofilms and antifungal resistance. In Alix T Coste, Vandeputte Patrick. (editors) Antifungals: from Genomics to Resistance and the Development of Novel Agents Norfolk, United Kingdom: Caister Academic Press; 2015 pp 71–90
    [Google Scholar]
  20. den Hertog AL, van Marle J, van Veen HA, Van't Hof W, Bolscher JGM et al. Candidacidal effects of two antimicrobial peptides: histatin 5 causes small membrane defects, but LL-37 causes massive disruption of the cell membrane. Biochem J 2005; 388:689–695 [CrossRef][PubMed]
    [Google Scholar]
  21. Tsai P-W, Yang C-Y, Chang H-T, Lan C-Y. Characterizing the role of cell-wall β-1,3-exoglucanase Xog1p in Candida albicans adhesion by the human antimicrobial peptide LL-37. PLoS One 2011; 6:e21394 [CrossRef][PubMed]
    [Google Scholar]
  22. Dumoulin M, Johnson RJ, Bellotti V, Dobson CM. Human lysozyme. In Uversky VN, Fink A. (editors) Protein Misfolding, Aggregation, and Conformational Diseases Boston: Springer; 2007 pp 285–308
    [Google Scholar]
  23. Humann J, Lenz LL. Bacterial peptidoglycan degrading enzymes and their impact on host muropeptide detection. J Innate Immun 2009; 1:88–97 [CrossRef][PubMed]
    [Google Scholar]
  24. Sherry L, Rajendran R, Lappin DF, Borghi E, Perdoni F et al. Biofilms formed by Candida albicans bloodstream isolates display phenotypic and transcriptional heterogeneity that are associated with resistance and pathogenicity. BMC Microbiol 2014; 14:182 [CrossRef][PubMed]
    [Google Scholar]
  25. Wayne P. Clinical and laboratory Standards Institute: reference method for broth dilution antifungal susceptibility testing of yeasts; Approved standard. CLSI document 2012M27–S4
    [Google Scholar]
  26. Moody JA. Synergism testing. Broth microdilution checkerboard and broth microdilution methods. In Isenberg HD. editor Clinical Microbiology Pro- Cedures Handbook 1 Washington, DC: Mbio; 1991 pp 1–28
    [Google Scholar]
  27. Incani V, Omar A, Prosperi-Porta G, Nadworny P. Ag5IO6: novel antibiofilm activity of a silver compound with application to medical devices. Int J Antimicrob Agents 2015; 45:586–593 [CrossRef][PubMed]
    [Google Scholar]
  28. Harrison JJ, Stremick CA, Turner RJ, Allan ND, Olson ME et al. Microtiter susceptibility testing of microbes growing on peg lids: a miniaturized biofilm model for high-throughput screening. Nat Protoc 2010; 5:1236–1254 [CrossRef][PubMed]
    [Google Scholar]
  29. Valentín A, Cantón E, Pemán J, Fernandez-Rivero ME, Tormo-Mas MA et al. In vitro activity of anidulafungin in combination with amphotericin B or voriconazole against biofilms of five Candida species. J Antimicrob Chemother 2016; 71:3449–3452 [CrossRef][PubMed]
    [Google Scholar]
  30. Pierce CG, Uppuluri P, Tristan AR, Wormley FL, Mowat E et al. A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing. Nat Protoc 2008; 3:1494–1500 [CrossRef]
    [Google Scholar]
  31. Lohse MB, Gulati M, Johnson AD, Nobile CJ. Development and regulation of single- and multi-species Candida albicans biofilms. Nat Rev Microbiol 2018; 16:19–31 [CrossRef]
    [Google Scholar]
  32. Soldini S, Posteraro B, Vella A, De Carolis E, Borghi E et al. Microbiologic and clinical characteristics of biofilm-forming Candida parapsilosis isolates associated with fungaemia and their impact on mortality. Clin Microbiol Infect 2018; 24:771–777 [CrossRef]
    [Google Scholar]
  33. Liao Y, Yang S, Cong L, Lu X, Ao J et al. In Vitro Activities of Antifungal Combinations against Biofilms and Planktonic Forms of Clinical Trichosporon asahii Isolates. Antimicrob Agents Chemother 2014; 58:7615–7616 [CrossRef]
    [Google Scholar]
  34. De Cremer K, Staes I, Delattin N, Cammue BPA, Thevissen K et al. Combinatorial drug approaches to tackle Candida albicans biofilms. Expert Rev Anti Infect Ther 2015; 13:973–984 [CrossRef]
    [Google Scholar]
  35. Nett J, Lincoln L, Marchillo K, Massey R, Holoyda K et al. Putative role of β-1,3 glucans in Candida albicans biofilm resistance. Antimicrob Agents Chemother 2007; 51:510–520 [CrossRef]
    [Google Scholar]
  36. Rodrigues C, Alves D, Henriques M. Combination of Posaconazole and Amphotericin B in the Treatment of Candida glabrata Biofilms. Microorganisms 2018; 6:123 [CrossRef]
    [Google Scholar]
  37. Mohammad H, Eldesouky HE, Hazbun T, Mayhoub AS, Seleem MN. Identification of a Phenylthiazole Small Molecule with Dual Antifungal and Antibiofilm Activity Against Candida albicans and Candida auris. Sci Rep 2019; 9:18941 [CrossRef]
    [Google Scholar]
  38. Samaranayake YH, Cheung BPK, Parahitiyawa N, Seneviratne CJ, Yau JYY et al. Synergistic activity of lysozyme and antifungal agents against Candida albicans biofilms on denture acrylic surfaces. Arch Oral Biol 2009; 54:115–126 [CrossRef]
    [Google Scholar]
  39. Sebaa S, Hizette N, Boucherit-Otmani Z, Courtois P. Dose-Dependent effect of lysozyme upon Candida albicans biofilm. Mol Med Rep 2017; 15:1135–1142 [CrossRef]
    [Google Scholar]
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