Skip to main content
SpringerLink
Log in

Broth Microdilution and Time–Kill Testing of Caspofungin, Voriconazole, Amphotericin B and their Combinations Against Clinical Isolates of Candida krusei

  • Published:
Mycopathologia Aims and scope Submit manuscript

Abstract

Treatment of invasive Candida krusei infections can be difficult due to its intrinsic fluconazole resistance and its reduced susceptibility to amphotericin B and flucytosine. Caspofungin (CAS) acts on a different cellular target, and its combination with voriconazole (VOR) or amphotericin B (AmB) appears promising. We evaluated the activity of CAS, VOR and AmB alone and in combination at 1/4, 1, 4xMIC concentrations by time–kill method against 30 C. krusei isolates. All isolates were susceptible to CAS and VOR; AmB MICs were 2 μg/ml for 50% of isolates by broth microdilution. CAS showed a fast killing activity at all concentrations; it was fungistatic at 1/4xMICs and fungicidal at 1-4xMICs in general. VOR displayed a concentration-independent fungistatic activity against all isolates. AmB exhibited a concentration-dependent activity; it was fungistatic at 1/4-1xMIC and fungicidal at 4xMIC. The most common interaction was indifference for both combinations. Frequency of synergic interaction for the VOR + CAS combination was 66.7% at 1/4xMIC after 48 h. The best results for CAS + AmB combination were obtained at 4xMIC in the first 4–8 h; synergic interaction was detected for 20 isolates (66.7%) at 4xMIC after 4 h. Consequently, VOR and CAS alone have been found effective, and high AmB MICs are remarkable against clinical C. krusei isolates in vitro. The combinations of CAS with VOR or AmB have exhibited promising results.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  1. Mukherjee PK, Sheehan DJ, Hitchcock CA, Ghannoum MA. Combination treatment of invasive fungal infections. Clin Microbiol Rev. 2005;18:163–94.

    Article  PubMed  CAS  Google Scholar 

  2. Howden BP, Slavin MA, Schwarer AP, Mijch AM. Successful control of disseminated Scedosporium prolificans infection with a combination of voriconazole and terbinafine. Eur J Clin Microbiol Infect Dis. 2003;22:111–3.

    PubMed  CAS  Google Scholar 

  3. Park D, Sohn J, Cheong H, Kim W, Ja Kim M, Kim JH, Shin C. Combination therapy of disseminated coccidioidomycosis with caspofungin and fluconazole. BMC Infect Dis. 2006;6:26.

    Article  PubMed  Google Scholar 

  4. Chaturvedi V, Ramani R, Ghannoum MA, Killian SB, Holliday N, Knapp C, Ostrosky-Zeichner L, Messer SA, Pfaller MA, Iqbal NJ, Arthington-Skaggs BA, Vazquez JA, Sein T, Rex JH, Walsh TJ. Multilaboratory testing of antifungal combinations against a quality control isolate of Candida krusei. Antimicrob Agents Chemother. 2008;52:1500–2.

    Article  PubMed  CAS  Google Scholar 

  5. Pfaller MA, Diekema DJ. Epidemiology of invasive candidiasis: a persistent public health problem. Clin Microbiol Rev. 2007;20:133–63.

    Article  PubMed  CAS  Google Scholar 

  6. Hachem R, Hanna H, Kontoyiannis D, Jiang Y, Raad I. The changing epidemiology of invasive candidiasis: Candida glabrata and Candida krusei as the leading causes of candidemia in hematologic malignancy. Cancer. 2008;112:2493–9.

    Article  PubMed  Google Scholar 

  7. Pfaller MA, Diekema DJ, Gibbs DL, Newell VA, Nagy E, Dobiasova S, Rinaldi M, Barton R, Veselov A. Global antifungal Surveillance group. Candida krusei, a multidrug-resistant opportunistic fungal pathogen: geographic and temporal trends from the ARTEMIS DISK antifungal surveillance program, 2001 to 2005. J Clin Microbiol. 2008;46:515–21.

    Article  PubMed  CAS  Google Scholar 

  8. Canton E, Pemán J, Valentín A, Espinel-Ingroff A, Gobernado M. In vitro activities of echinocandins against Candida krusei determined by three methods: MIC and minimal fungicidal concentration measurements and time-kill studies. Antimicrob Agents Chemother. 2009;53:3108–11.

    Article  PubMed  CAS  Google Scholar 

  9. Clinical and Laboratory Standards Institute. Reference method for broth dilution antifungal susceptibility testing of yeast; approved Standard-third edition. Document M27-A3. Clinical and Laboratory Standards Institute, 2008; Wayne, PA.

  10. Pfaller MA, Diekema DJ, Rex JH, Espinel-Ingroff A, Johnson EM, Andes D, Chaturvedi V, Ghannoum MA, Odds FC, Rinaldi MG, Sheehan DJ, Troke P, Walsh TJ, Warnock DW. Correlation of MIC with outcome for Candida species tested against voriconazole: analysis and proposal for interpretive breakpoints. J Clin Microbiol. 2006;44:819–26.

    Article  PubMed  CAS  Google Scholar 

  11. Canton E, Espinel-Ingroff A, Pemán J. Trends in antifungal susceptibility testing using CLSI reference and commercial methods. Expert Rev Anti Infect Ther. 2009;7:107–19.

    Article  PubMed  CAS  Google Scholar 

  12. Pfaller MA, Diekema DJ, Ostrosky-Zeichner L, Rex JH, Alexander BD, Andes D, Brown SD, Chaturvedi V, Ghannoum MA, Knapp CC, Sheehan DJ, Walsh TJ. Correlation of MIC with outcome for Candida species tested against caspofungin, anidulafungin, and micafungin: analysis and proposal for interpretive MIC breakpoints. J Clin Microbiol. 2008;46:2620–9.

    Article  PubMed  CAS  Google Scholar 

  13. González GM, Elizondo M, Ayala J. Trends in species distribution and susceptibility of bloodstream isolates of Candida collected in Monterrey, Mexico, to seven antifungal agents: results of a 3-year (2004 to 2007) surveillance study. J Clin Microbiol. 2008;46:2902–5.

    Article  PubMed  Google Scholar 

  14. Negri M, Henriques M, Svidzinski TI, Paula CR, Oliveira R. Correlation between Etest, disk diffusion, and microdilution methods for antifungal susceptibility testing of Candida species from infection and colonization. J Clin Lab Anal. 2009;23:324–30.

    Article  PubMed  Google Scholar 

  15. Klepser ME, Wolfe EJ, Jones RN, Nightingale CH, Pfaller MA. Antifungal pharmacodynamic characteristics of fluconazole and amphotericin B tested against Candida albicans. Antimicrob Agents Chemother. 1997;41:1392–5.

    PubMed  CAS  Google Scholar 

  16. Lewis RE, Diekema DJ, Messer SA, Pfaller MA, Klepser ME. Comparison of Etest, chequerboard dilution and time-kill studies for the detection of synergy or antagonism between antifungal agents tested against Candida species. J Antimicrob Chemother. 2002;49:345–51.

    Article  PubMed  CAS  Google Scholar 

  17. Klepser ME, Ernst EJ, Lewis RE, Ernst ME, Pfaller MA. Influence of test conditions on antifungal time-kill curve results: proposal for standardized methods. Antimicrob Agents Chemother. 1998;42:1207–12.

    PubMed  CAS  Google Scholar 

  18. Roling EE, Klepser ME, Wasson A, Lewis RE, Ernst EJ, Pfaller MA. Antifungal activities of fluconazole, caspofungin (MK0991), and anidulafungin (LY 303366) alone and in combination against Candida spp. and Cryptococcus neoformans via time-kill methods. Diagn Microbiol Infect Dis. 2002;43:13–7.

    Article  PubMed  CAS  Google Scholar 

  19. Di Bonaventura G, Spedicato I, Picciani C, D’Antonio D, Piccolomini R. In vitro pharmacodynamic characteristics of amphotericin B, caspofungin, fluconazole, and voriconazole against bloodstream isolates of infrequent Candida species from patients with hematologic malignancies. Antimicrob Agents Chemother. 2004;48:4453–6.

    Google Scholar 

  20. Szabo Z, Borbely A, Kardos G, Somogyvari F, Kemény-Beke A, Asztalos L, Rozgonyi F, Majoros L. In vitro efficacy of amphotericin B 5-fluorocytosine, fluconazole, voriconazole and posaconazole against Candida dubliniensis isolates using time-kill methodology. Mycoses. 2010;53:196–9.

    Article  PubMed  CAS  Google Scholar 

  21. Barchiesi F, Spreghini E, Tomassetti S, Arzeni D, Giannini D, Scalise G. Comparison of the fungicidal activities of caspofungin and amphotericin B against Candida glabrata. Antimicrob Agents Chemother. 2005;49:4989–92.

    Article  PubMed  CAS  Google Scholar 

  22. Canton E, Peman J, Gobernado M, Viudes A, Espinel-Ingroff A. Patterns of amphotericin B killing kinetics against seven Candida species. Antimicrob Agents Chemother. 2004;48:2477–82.

    Article  PubMed  CAS  Google Scholar 

  23. Baltch AL, Bopp LH, Smith RP, Ritz WJ, Michelsen PB. Anticandidal effects of voriconazole and caspofungin, singly and in combination, against Candida glabrata, extracellularly and intracellularly in granulocyte-macrophage colony stimulating factor (GM-CSF)-activated human monocytes. J Antimicrob Chemother. 2008;62:1285–90.

    Article  PubMed  CAS  Google Scholar 

  24. MacCallum DM, Whyte JA, Odds FC. Efficacy of caspofungin and voriconazole combinations in experimental aspergillosis. Antimicrob Agents Chemother. 2005;49:3697–701.

    Article  PubMed  CAS  Google Scholar 

  25. Steinbach WJ, Stevens DA, Denning DW. Combination and sequential antifungal therapy for invasive aspergillosis: review of published in vitro and in vivo interactions and 6281 clinical cases from 1966 to 2001. Clin Infect Dis. 2003;37:S188–224.

    Article  PubMed  CAS  Google Scholar 

  26. Dannaoui E, Lortholary O, Dromer F. In vitro evaluation of double and triple combinations of antifungal drugs against Aspergillus fumigatus and Aspergillus terreus. Antimicrob Agents Chemother. 2004;48:970–8.

    Article  PubMed  CAS  Google Scholar 

  27. Li H, Lu Q, Wan Z, Zhang J. In vitro combined activity of amphotericin B, caspofungin and voriconazole against clinical isolates of Trichosporon asahii. Int J Antimicrob Agents. 2010;35:550–2.

    Article  PubMed  CAS  Google Scholar 

  28. Kiraz N, Dag I, Yamac M, Kiremitci A, Kasifoglu N, Akgun Y. Antifungal activity of caspofungin in combination with amphotericin B against Candida glabrata: comparison of disk diffusion, Etest, and time-kill methods. Antimicrob Agents Chemother. 2009;53:788–90.

    Article  PubMed  CAS  Google Scholar 

  29. Kiraz N, Dag I, Yamac M, Kiremitci A, Kasifoglu N, Oz Y. Synergistic activities of three triazoles with caspofungin against Candida glabrata isolates determined by time-Kill, Etest and disk diffusion methods. Antimicrob Agents Chemother. 2010;54:2244–7.

    Article  PubMed  CAS  Google Scholar 

  30. Barchiesi F, Spreghini E, Tomassetti S, Giannini D, Scalise G. Caspofungin in combination with amphotericin B against Candida parapsilosis. Antimicrob Agents Chemother. 2007;51:941–5.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by a grant of Turkey Scientific and Technological Research Foundation-TUBITAK (No. 109S015).

Author information

Authors and Affiliations

  1. Department of Microbiology, Division of Mycology, Eskisehir Osmangazi University Medical Faculty, Eskisehir, Turkey

    Yasemin Oz

  2. Health Services Vocational High School, Eskisehir Osmangazi University, Eskisehir, Turkey

    Ilknur Dag

  3. Department of Microbiology, Division of Mycology, Istanbul University Cerrahpasa Medical Faculty, Istanbul, Turkey

    Nuri Kiraz

Authors
  1. Yasemin Oz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ilknur Dag
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nuri Kiraz
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yasemin Oz.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Oz, Y., Dag, I. & Kiraz, N. Broth Microdilution and Time–Kill Testing of Caspofungin, Voriconazole, Amphotericin B and their Combinations Against Clinical Isolates of Candida krusei . Mycopathologia 173, 27–34 (2012). https://doi.org/10.1007/s11046-011-9459-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11046-011-9459-x

Keywords

Search

Navigation