Abstract
Fungal biofilms formed on various types of medical implants represent a major problem for hospitalized patients. These biofilms and related infections are usually difficult to treat because of their resistance to the classical antifungal drugs. Animal models are indispensable for investigating host–pathogen interactions and for identifying new antifungal targets related to biofilm development. A limited number of animal models is available that can be used for testing novel antifungal drugs in vivo against C. albicans, one of the most common pathogens causing fungal biofilms. Fungal load in biofilms in these models is traditionally analyzed postmortem, requiring host sacrifice and enumeration of microorganisms from individual biofilms in order to evaluate the amount of colony forming units and the efficacy of antifungal treatment. Bioluminescence imaging (BLI) made compatible with small animal models for in vivo biofilm formation is a valuable noninvasive tool to follow-up biofilm development and its treatment longitudinally, reducing the number of animals needed for such studies. Due to the nondestructive and noninvasive nature of BLI, the imaging procedure can be repeated in the same animal, allowing follow-up of the biofilm growth in vivo without removing the implanted device or detaching the biofilm from its substrate. The method described here introduces BLI of C. albicans biofilm formation in vivo on subcutaneously implanted catheters in mice. One of the main challenges to overcome for BLI of fungi is the hampered intracellular substrate delivery through the fungal cell wall, which is managed by using extracellularly located Gaussia luciferase. Although detecting a quantifiable in vivo BLI signal from biofilms formed on the inside of implanted catheters is challenging, BLI proved to be a practical tool in the study of fungal biofilms. This method describing the use of BLI for in vivo follow-up of device-related fungal biofilm formation has the potential for efficient in vivo screening for interesting genes of the pathogen and the host involved in C. albicans biofilm formation as well as for testing novel antifungal therapies.
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References
Warnock DW (2006) Fungal diseases: an evolving public health challenge. Med Mycol 44(8):697–705
Nucci M, Marr KA (2005) Emerging fungal diseases. Clin Infect Dis 41(4):521–526
Pfaller MA, Diekema DJ (2010) Epidemiology of invasive mycoses in North America. Crit Rev Microbiol 36(1):1–53
Eggimann P, Garbino J, Pittet D (2003) Management of Candida species infections in critically ill patients. Lancet Infect Dis 3(12):772–785
Eggimann P, Garbino J, Pittet D (2003) Epidemiology of Candida species infections in critically ill non-immunosuppressed patients. Lancet Infect Dis 3(11):685–702
Pemán J, Salavert M (2012) Epidemiología general de la enfermedad fúngica invasora. Enferm Infecc Microbiol Clin 30(2):90–98
Wisplinghoff H, Bischoff T, Tallent SM, Seifert H, Wenzel RP, Edmond MB (2004) Nosocomial bloodstream infections in US hospitals: analysis of 24,179 cases from a prospective nationwide surveillance study. Clin Infect Dis 39(3):309–317
Enoch DA, Ludlam HA, Brown NM (2006) Invasive fungal infections: a review of epidemiology and management options. J Med Microbiol 55(Pt 7):809–818
Nett J, Andes D (2006) Candida albicans biofilm development, modeling a host-pathogen interaction. Curr Opin Microbiol 9(4):340–345
Andes D, Nett J, Oschel P, Albrecht R, Marchillo K, Pitula A (2004) Development and characterization of an in vivo central venous catheter Candida albicans biofilm model. Infect Immun 72(10):6023–6031
Schinabeck MK, Long LA, Hossain MA, Chandra J, Mukherjee PK, Mohamed S, Ghannoum MA (2004) Rabbit model of Candida albicans biofilm infection: liposomal amphotericin B antifungal lock therapy. Antimicrob Agents Chemother 48(5):1727–1732
Lazzell AL, Chaturvedi AK, Pierce CG, Prasad D, Uppuluri P, Lopez-Ribot JL (2009) Treatment and prevention of Candida albicans biofilms with caspofungin in a novel central venous catheter murine model of candidiasis. J Antimicrob Chemother 64(3):567–570
Van Wijngaerden E, Peetermans WE, Vandersmissen J, Van Lierde S, Bobbaers H, Van Eldere J (1999) Foreign body infection: a new rat model for prophylaxis and treatment. J Antimicrob Chemother 44(5):669–674
Rˇicˇicová M, Kucharíková S, Tournu H, Hendrix J, Bujdáková H, Van Eldere J, Lagrou K, Van Dijck P (2009) Candida albicans biofilm formaton in a new in vivo rat model. Microbiology 156(Pt3):909–919
Kucharíková S, Tournu H, Holtappels M, Van Dijck P, Lagrou K (2010) In vivo efficacy of anidulafungin against mature Candida albicans biofilms in a novel rat model of catheter-associated Candidiasis. Antimicrob Agents Chemother 54(10):4474–4475
Hutchens M, Luker GD (2007) Applications of bioluminescence imaging to the study of infectious diseases. Cell Microbiol 9(10):2315–2322
Doyle TC, Nawotka KA, Kawahara CB, Francis KP, Contag PR (2006) Visualizing fungal infections in living mice using bioluminescent pathogenic Candida albicans strains transformed with the firefly luciferase gene. Microb Pathog 40(2):82–90
Enjalbert B, Rachini A, Vediyappan G, Pietrella D, Spaccapelo R, Vecchiarelli A, Brown AJ, d'Enfert C (2009) A multifunctional, synthetic Gaussia princeps luciferase reporter for live imaging of Candida albicans infections. Infect Immun 77(11):4847–4858
Gillum AM, Tsay EY, Kirsch DR (1984) Isolation of the Candida albicans gene for orotidine-5'-phosphate decarboxylase by complementation of S. cerevisiae ura3 and E. coli pyrF mutations. Mol Gen Genet 198(1):179–182
Acknowledgements
This work was funded by KU Leuven PF “IMIR,” the FWO Research community on biology and ecology of bacterial and fungal biofilms (FWO: WO.026.11N), KU Leuven PDMK 11/089 fellowship and FWO postdoctoral fellowship to SK. We thank Christophe d’Enfert for providing us with the Clp10::ACT1p-gLUC59 plasmid.
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Velde, G.V., Kucharíková, S., Van Dijck, P., Himmelreich, U. (2014). Bioluminescence Imaging of Fungal Biofilm Development in Live Animals. In: Badr, C. (eds) Bioluminescent Imaging. Methods in Molecular Biology, vol 1098. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-718-1_13
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DOI: https://doi.org/10.1007/978-1-62703-718-1_13
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