Skip to main content

Advertisement

SpringerLink
Log in

Murine Model of Disseminated Fusariosis: Evaluation of the Fungal Burden by Traditional CFU and Quantitative PCR

  • Published:
Mycopathologia Aims and scope Submit manuscript

Abstract

Systemic disease is the most severe clinical form of fusariosis, and the treatment involves a challenge due to the refractory response to antifungals. Treatment for murine Fusarium solani infection has been described in models that employ CFU quantitation in organs as a parameter of therapeutic efficacy. However, CFU counts do not precisely reproduce the amount of cells for filamentous fungi such as F. solani. In this study, we developed a murine model of disseminated fusariosis and compared the fungal burden with two methods: CFU and quantitative PCR. ICR and BALB/c mice received an intravenous injection of 1 × 107 conidia of F. solani per mouse. On days 2, 5, 7, and 9, mice from each mice strain were killed. The spleen and kidneys of each animal were removed and evaluated by qPCR and CFU determinations. Results from CFU assay indicated that the spleen and kidneys had almost the same fungal burden in both BALB/c and ICR mice during the days of the evaluation. In the qPCR assay, the spleen and kidney of each mouse strain had increased fungal burden in each determination throughout the entire experiment. The fungal load determined by the qPCR assay was significantly greater than that determined from CFU measurements of tissue. qPCR could be considered as a tool for quantitative evaluation of fungal burden in experimental disseminated F. solani infection.

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
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Nucci M, Anaissie E. Fusarium infections in immunocompromised patients. Clin Microbiol Rev. 2007;20:695–704.

    Article  PubMed  CAS  Google Scholar 

  2. Dignani MC, Anaissie E. Human fusariosis. Clin Microbiol Infect Dis. 2004;10(suppl 1):67–75.

    Article  Google Scholar 

  3. Martino P, Gastaldi R, Raccah R, Girmenia C. Clinical patterns of Fusarium infections in immunocompromised patients. J Infect. 1994;28(suppl 1):7–15.

    Article  PubMed  Google Scholar 

  4. Walsh TJ, Groll AH. Emerging fungal pathogens: evolving challenges to immunocompromised patients for the twenty-first century. Transpl Infect Dis. 1999;1:247–61.

    Article  PubMed  CAS  Google Scholar 

  5. Girmenia C, Pagano L, Corvatta L, Mele L, del Favero A, Martino P. The epidemiology of fusariosis in patients with haematological diseases. Br J Haematol. 2000;111:272–6.

    Article  PubMed  CAS  Google Scholar 

  6. Pontón J, Rüchel R, Clemons KV, Coleman DC, Grillot R, Guarro J, Aldebert D, Ambroise-Thomas P, Cano J, Carrillo-Muñoz AJ, Gené J, Pinel C, Stevens DA, Sullivan DJ. Emerging pathogens. Med Mycol. 2000;38(suppl 1):S225–36.

    Google Scholar 

  7. Mayayo E, Pujol I, Guarro J. Experimental pathogenicity of four opportunist Fusarium species in a murine model. J Med Microbiol. 1999;48:363–6.

    Article  PubMed  CAS  Google Scholar 

  8. Azor M, Gene J, Cano J, Guarro J. Universal in vitro antifungal resistance of genetic clades of the Fusarium solani species complex. Antimicrob Agents Chemother. 2007;51:1500–3.

    Article  PubMed  CAS  Google Scholar 

  9. Alastruey-Izquierdo A, Cuenca-Estrella M, Monzón A, Mellado E, Rodríguez-Tudela JL. Antifungal susceptibility profile of clinical Fusarium spp. Isolates identified by molecular methods. J Antimicrob Chemother. 2008;61:805–9.

    Article  PubMed  CAS  Google Scholar 

  10. O′Donnell K, Sutton DA, Fothergill A, McCarthy D, Rinaldi MG, Brandt ME, Zhang N, Geiser DM. Molecular phylogenetic, multilocus haplotype nomenclature, and in vitro antifungal resistance within the Fusarium solani species complex. J Clin Microbiol. 2008;46:2477–90.

    Article  PubMed  Google Scholar 

  11. Nucci M, Marr KA, Queiroz-Telles F, Martins CA, Trabasso P, Costa S, Voltarelli JC, Colombo AL, Imhof A, Pasquini R, Maiolino A, Souza CA, Anaissie E. Fusarium infection in hematopoietic stem cell transplant recipients. Clin Infect Dis. 2004;38:1237–42.

    Article  PubMed  Google Scholar 

  12. Perfect JR, Marr KA, Walsh TJ, Greenberg RN, DuPont B, de la Torre-Cisneros J, Just-Nübling G, Schlamm HT, Lutsar I, Espinel-Ingroff A, Johnson E. Voriconazole treatment for less-common, emerging, or refractory fungal infections. Clin Infect Dis. 2003;36:1122–31.

    Article  PubMed  CAS  Google Scholar 

  13. Raad II, Hachem RY, Herbrecht R, Graybill JR, Hare R, Corcoran G, Kontoyiannis DP. Posaconazole as salvage treatment for invasive fusariosis in patients with underlying hematologic malignancy and other conditions. Clin Infect Dis. 2006;42:1398–403.

    Article  PubMed  CAS  Google Scholar 

  14. Lozano-Chiu M, Arikan S, Paetznick VL, Anaissie EJ, Loebenberg D, Rex JH. Treatment of murine fusariosis with SCH56592. Antimicrob Agents Chemother. 1999;43:589–91.

    Article  PubMed  CAS  Google Scholar 

  15. Spellberg B, Schwartz J, Fu Y, Avanesian V, Adler-Moore J, Edwards JE Jr, Ibrahim AS. Comparison of antifungal treatments for murine fusariosis. J Antimicrob Chemother. 2006;58:973–9.

    Article  PubMed  CAS  Google Scholar 

  16. Ruíz-Cendoya M, Mariné M, Guarro J. Combined therapy in treatment of murine infection by Fusarium solani. J Antimicrob Chemother. 2008;62:543–6.

    Article  PubMed  Google Scholar 

  17. Wiederhold NP, Najvar LK, Bocanegra R, Graybill JR, Patterson TF. Efficacy of posaconazole as treatment and prophylaxis against Fusarium solani. Antimicrob Agents Chemother. 2010;54:1055–9.

    Article  PubMed  CAS  Google Scholar 

  18. Nelson PE, Tousson TA, Marasas WFO. Fusarium species: an illustrated manual for identification. University Park, PA: Pennsylvania State University Press; 1983.

    Google Scholar 

  19. Rosa RH Jr, Miller D, Alfonso EC. The changing spectrum of fungal keratitis in south Florida. Ophthalmology. 1994;101:1005–13.

    PubMed  Google Scholar 

  20. Doyle JJ, Doyle JL. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull. 1987;19:11–5.

    Google Scholar 

  21. Cullings KW. Design and testing of a plant-specific PCR primer for ecological and evolutionary studies. Mol Ecol. 1992;1:233–40.

    Article  CAS  Google Scholar 

  22. Jaeger EE, Carroll NM, Choudhury S, Dunlop AA, Towler HM, Matheson MM, Adamson P, Okhravi N, Lightman S. Rapid detection and identification of Candida, Aspergillus, and Fusarium species in ocular samples using nested PCR. J Clin Microbiol. 2000;38:2902–8.

    PubMed  CAS  Google Scholar 

  23. Walsh TJ, McEntee C, Dixon DM. Tissue homogenization with sterile reinforced polyethylene bags for quantitative culture of Candida albicans. J Clin Microbiol. 1987;25:931–2.

    PubMed  CAS  Google Scholar 

  24. Latgé JP. Aspergillus fumigatus and aspergillosis. Clin Microbiol Rev. 1999;12:310–50.

    PubMed  Google Scholar 

Download references

Acknowledgements

We thank Sergio Lozano-Rodriguez, M.D. for his review of the manuscript prior to submission.

Conflict of interest

The authors declare that no conflict of interest.

Author information

Authors and Affiliations

  1. Departamento de Microbiología, Facultad de Medicina, Universidad Autónoma de Nuevo León, Madero y Dr. Eduardo A. Pequeño s/n, Colonia Mitras Centro, 64460, Monterrey, NL, Mexico

    Gloria M. González, Jazmín Márquez, Rogelio de J. Treviño-Rangel, José P. Palma-Nicolás & Elvira Garza-González

  2. Facultad de Medicina, Hospital Universitario, Universidad Autónoma de Nuevo León, Monterrey, Mexico

    Luis A. Ceceñas & J. Gerardo González

Authors
  1. Gloria M. González
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jazmín Márquez
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rogelio de J. Treviño-Rangel
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. José P. Palma-Nicolás
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Elvira Garza-González
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Luis A. Ceceñas
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J. Gerardo González
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Rogelio de J. Treviño-Rangel.

Rights and permissions

Reprints and permissions

About this article

Cite this article

González, G.M., Márquez, J., Treviño-Rangel, R.J. et al. Murine Model of Disseminated Fusariosis: Evaluation of the Fungal Burden by Traditional CFU and Quantitative PCR. Mycopathologia 176, 219–224 (2013). https://doi.org/10.1007/s11046-013-9687-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11046-013-9687-3

Keywords

Search

Navigation