Skip to main content

Advertisement

SpringerLink
Log in

Candida tropicalis Biofilms: Biomass, Metabolic Activity and Secreted Aspartyl Proteinase Production

  • Published:
Mycopathologia Aims and scope Submit manuscript

Abstract

According to epidemiological data, Candida tropicalis has been related to urinary tract infections and haematological malignancy. Several virulence factors seem to be responsible for C. tropicalis infections, for example: their ability to adhere and to form biofilms onto different indwelling medical devices; their capacity to adhere, invade and damage host human tissues due to enzymes production such as proteinases. The main aim of this work was to study the behaviour of C. tropicalis biofilms of different ages (24–120 h) formed in artificial urine (AU) and their ability to express aspartyl proteinase (SAPT) genes. The reference strain C. tropicalis ATCC 750 and two C. tropicalis isolates from urine were used. Biofilms were evaluated in terms of culturable cells by colony-forming units enumeration; total biofilm biomass was evaluated using the crystal violet staining method; metabolic activity was evaluated by XTT assay; and SAPT gene expression was determined by real-time PCR. All strains of C. tropicalis were able to form biofilms in AU, although with differences between strains. Candida tropicalis biofilms showed a decrease in terms of the number of culturable cells from 48 to 72 h. Generally, SAPT3 was highly expressed. C. tropicalis strains assayed were able to form biofilms in the presence of AU although in a strain- and time-dependent way, and SAPT genes are expressed during C. tropicalis biofilm formation.

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. Sardi J, Scorzoni L, Bernardi T, Fusco-Almeida A, Giannini MM. Candida species: current epidemiology, pathogenicity, biofilm formation, natural antifungal products and new therapeutic options. J Med Microbiol. 2013;62:10–24.

    Article  CAS  PubMed  Google Scholar 

  2. Lass-Flörl C. The changing face of epidemiology of invasive fungal disease in Europe. Mycoses. 2009;52:197–205.

    Article  PubMed  Google Scholar 

  3. Falagas ME, Roussos N, Vardakas KZ. Relative frequency of albicans and the various non-albicans Candida spp among candidemia isolates from inpatients in various parts of the world: a systematic review. Int J Infect Dis. 2010;14:954–66.

    Article  Google Scholar 

  4. Nucci M, Queiroz-Telles F, Tobón AM, Restrepo A, Colombo AL. Epidemiology of opportunistic fungal infections in Latin America. Clin Infect Dis. 2010;51:561–70.

    Article  PubMed  Google Scholar 

  5. Horn DL, Neofytos D, Anaissie EJ, Fishman JA, Steinbach WJ, Olyaei AJ, Marr KA, Pfaller MA, Chang C-H, Webster KM. Epidemiology and outcomes of candidemia in 2019 patients: data from the prospective antifungal therapy alliance registry. Clin Infect Dis. 2009;48:1695–703.

    Article  CAS  PubMed  Google Scholar 

  6. Paul N, Mathai E, Abraham OC, Michael JS, Mathai D. Factors associated with candiduria and related mortality. J Infect. 2007;55:450–5.

    Article  PubMed  Google Scholar 

  7. Negri M, Silva S, Breda D, Henriques M, Azeredo J, Oliveira R. Candida tropicalis biofilms: effect on urinary epithelial cells. Microb Pathog. 2012;53:95–9.

    Article  CAS  PubMed  Google Scholar 

  8. Goetz LL, Howard M, Cipher D, Revankar SG. Occurrence of candiduria in a population of chronically catheterized patients with spinal cord injury. Spinal Cord. 2009;48:51–4.

    Article  PubMed  Google Scholar 

  9. Kothavade RJ, Kura MM, Valand AG, Panthaki MH. Candida tropicalis: its prevalence, pathogenicity and increasing resistance to fluconazole. J Med Microbiol. 2010;59:873–80.

    Article  CAS  PubMed  Google Scholar 

  10. Munoz P, Giannella M, Fanciulli C, Guinea J, Valerio M, Rojas L, Rodríguez-Créixems M, Bouza E. Candida tropicalis fungaemia: incidence, risk factors and mortality in a general hospital. Clin Microbiol Infect. 2011;17:1538–45.

    Article  CAS  PubMed  Google Scholar 

  11. Silva S, Negri M, Henriques M, Oliveira R, Williams D, Azeredo J. Silicone colonization by non-Candida albicans Candida species in the presence of urine. J Med Microbiol. 2010;59:747–54.

    Article  CAS  PubMed  Google Scholar 

  12. Negri M, Silva S, Henriques M, Azeredo J, Svidzinski T, Oliveira R. Candida tropicalis biofilms: artificial urine, urinary catheters and flow model. Med Mycol. 2011;49:739–47.

    CAS  PubMed  Google Scholar 

  13. Deorukhkar SC, Saini S, Mathew S. Virulence factors contributing to pathogenicity of Candida tropicalis and its antifungal susceptibility profile. Int J Microbiol. 2014;2014:1–6.

    Article  Google Scholar 

  14. Silva S, Henriques M, Oliveira R, Williams D, Azeredo J. In vitro biofilm activity of non-Candida albicans Candida species. Curr Microbiol. 2010;61:534–40.

    Article  CAS  PubMed  Google Scholar 

  15. Negri M, Martins M, Henriques M, Svidzinski TI, Azeredo J, Oliveira R. Examination of potential virulence factors of Candida tropicalis clinical isolates from hospitalized patients. Mycopathologia. 2010;169:175–82.

    Article  CAS  PubMed  Google Scholar 

  16. Gokce G, Cerikcioglu N, Yagci A. Acid proteinase, phospholipase, and biofilm production of Candida species isolated from blood cultures. Mycopathologia. 2007;164:265–9.

    Article  CAS  PubMed  Google Scholar 

  17. Ramage G, Martínez JP, López-Ribot JL. Candida biofilms on implanted biomaterials: a clinically significant problem. FEMS Yeast Res. 2006;6:979–86.

    Article  CAS  PubMed  Google Scholar 

  18. Silva S, Negri M, Henriques M, Oliveira R, Williams DW, Azeredo J. Candida glabrata, Candida parapsilosis and Candida tropicalis: biology, epidemiology, pathogenicity and antifungal resistance. FEMS Microbiol Rev. 2012;36:288–305.

    Article  CAS  PubMed  Google Scholar 

  19. Kauffman CA, Vazquez JA, Sobel JD, Gallis HA, McKinsey DS, Karchmer AW, Sugar AM, Sharkey PK, Wise GJ, Mangi R. Prospective multicenter surveillance study of funguria in hospitalized patients. Clin Infect Dis. 2000;30:14–8.

    Article  CAS  PubMed  Google Scholar 

  20. Kojic EM, Darouiche RO. Candida infections of medical devices. Clin Microbiol Rev. 2004;17:255–67.

    Article  PubMed  PubMed Central  Google Scholar 

  21. Jain N, Kohli R, Cook E, Gialanella P, Chang T, Fries BC. Biofilm formation by and antifungal susceptibility of Candida isolates from urine. Appl Environ Microbiol. 2007;73:1697–703.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Bougnoux M-E, Kac G, Aegerter P, d’ Enfert C, Fagon J-Y. Candidemia and candiduria in critically ill patients admitted to intensive care units in France: incidence, molecular diversity, management and outcome. Intensive Care Med. 2008;34:292–9.

    Article  PubMed  Google Scholar 

  23. Achkar JM, Fries BC. Candida infections of the genitourinary tract. Clin Microbiol Rev. 2010;23:253–73.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Silva S, Negri M, Henriques M, Oliveira R, Williams DW, Azeredo J. Adherence and biofilm formation of non-Candida albicans Candida species. Trends Microbiol. 2011;19:241–7.

    Article  CAS  PubMed  Google Scholar 

  25. Nailis H, Coenye T, Van Nieuwerburgh F, Deforce D, Nelis HJ. Development and evaluation of different normalization strategies for gene expression studies in Candida albicans biofilms by real-time PCR. BMC Mol Biol. 2006;7:25.

    Article  PubMed  PubMed Central  Google Scholar 

  26. Marcos-Zambrano LJ, Escribano P, Bouza E, Guinea J. Production of biofilm by Candida and non-Candida spp. isolates causing fungemia: comparison of biomass production and metabolic activity and development of cut-off points. Int J Med Microbiol. 2014;304:1192–8.

    Article  CAS  PubMed  Google Scholar 

  27. Silva S, Hooper SJ, Henriques M, Oliveira R, Azeredo J, Williams DW. The role of secreted aspartyl proteinases in Candida tropicalis invasion and damage of oral mucosa. Clin Microbiol Infect. 2011;17:264–72.

    Article  CAS  PubMed  Google Scholar 

  28. Bizerra FC, Nakamura CV, De Poersch C, Estivalet Svidzinski TI, Borsato Quesada RM, Goldenberg S, Krieger MA, Yamada-Ogatta SF. Characteristics of biofilm formation by Candida tropicalis and antifungal resistance. FEMS Yeast Res. 2008;8:442–50.

    Article  CAS  PubMed  Google Scholar 

  29. Kuhn D, Balkis M, Chandra J, Mukherjee P, Ghannoum M. Uses and limitations of the XTT assay in studies of Candida growth and metabolism. J Clin Microbiol. 2003;41:506–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Costa AC, Pereira CA, Freire F, Junqueira JC, Jorge AO. Methods for obtaining reliable and reproducible results in studies of Candida biofilms formed in vitro. Mycoses. 2013;56:614–22.

    Article  PubMed  Google Scholar 

  31. Peeters E, Nelis HJ, Coenye T. Comparison of multiple methods for quantification of microbial biofilms grown in microtiter plates. J Microbiol Methods. 2008;72:157–65.

    Article  CAS  PubMed  Google Scholar 

  32. Negri M, Gonçalves VM, Silva SC, Henriques M, Azeredo J, Oliveira R. Crystal violet staining to quantity Candida adhesion to epithelial cells. Br J Biomed Sci. 2010;67:120–5.

    CAS  PubMed  Google Scholar 

  33. Negri M, Botelho C, Silva S, Lopes LM, Henriques M, Azeredo J, Oliveira R. An in vitro evaluation of Candida tropicalis infectivity using human cell monolayers. J Med Microbiol. 2011;60:1270–5.

    Article  CAS  PubMed  Google Scholar 

  34. Uppuluri P, Dinakaran H, Thomas DP, Chaturvedi AK, Lopez-Ribot JL. Characteristics of Candida albicans biofilms grown in a synthetic urine medium. J Clin Microbiol. 2009;47:4078–83.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Zaugg C, Borg-von Zepelin M, Reichard U, Sanglard D, Monod M. Secreted Aspartic Proteinase Family of Candida tropicalis. Infect Immun. 2001;69:405–12.

    Article  PubMed  PubMed Central  Google Scholar 

  36. Naglik JR, Challacombe SJ, Hube B. Candida albicans secreted aspartyl proteinases in virulence and pathogenesis. Microbiol Mol Biol Rev. 2003;67:400–28.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Togni G, Sanglard D, Falchetto R, Monod M. Isolation and nucleotide sequence of the extracellular acid protease gene (ACP) from the yeast Candida tropicalis. FEBS Lett. 1991;286:181–5.

    Article  CAS  PubMed  Google Scholar 

  38. Mendes A, Mores AU, Carvalho AP, Rosa RT, Samaranayake LP, Rosa EAR. Candida albicans biofilms produce more secreted aspartyl protease than the planktonic cells. Biol Pharm Bull. 2007;30:1813–5.

    Article  CAS  PubMed  Google Scholar 

  39. Nailis H, Kucharíková S, Řičicová M, Van Dijck P, Deforce D, Nelis H, Coenye T. Real-time PCR expression profiling of genes encoding potential virulence factors in Candida albicans biofilms: identification of model-dependent and -independent gene expression. BMC Microbiol. 2010;10:114.

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

The authors acknowledge Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brazil, for supporting Melyssa Negri (BEX 4642/06-6) and Fundação para a Ciência e Tecnologia (FCT), Portugal, for supporting Sonia Silva (SFRH/BPD/71076/2010) and European Community fund FEDER, through Program COMPETE, in the ambit of the Project FCOMP-01-0124-FEDER-007025 (PTDC/AMB/68393/2006).

Author contributions

MN conceived of the study, carried out the C. tropicalis biofilm studies, participated in the analysis of SAP gene expression and drafted the manuscript. SS carried out the analysis of SAP gene expression, performed the statistical analysis and helped to draft the manuscript. IRGC helped to draft the manuscript. JA participated in the design of the study and helped to draft the manuscript. MH participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.

Author information

Authors and Affiliations

  1. CEB- Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057, Braga, Portugal

    Melyssa Negri, Sónia Silva, Joana Azeredo & Mariana Henriques

  2. Teaching and Research in Clinical Analysis Laboratory, Division of Medical Mycology, Universidade Estadual de Maringá, Av. Colombo, 5790, Maringá, Paraná, 87020-900, Brazil

    Melyssa Negri & Isis Regina Grenier Capoci

Authors
  1. Melyssa Negri
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sónia Silva
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Isis Regina Grenier Capoci
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Joana Azeredo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mariana Henriques
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sónia Silva.

Ethics declarations

Conflict of interest

The authors declare that there is no conflict of interests regarding the publication of this paper.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Negri, M., Silva, S., Capoci, I.R.G. et al. Candida tropicalis Biofilms: Biomass, Metabolic Activity and Secreted Aspartyl Proteinase Production. Mycopathologia 181, 217–224 (2016). https://doi.org/10.1007/s11046-015-9964-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11046-015-9964-4

Keywords

Search

Navigation