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Biofilm formation and virulence properties of Candida spp. isolated from hospitalised patients in Tunisia

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Abstract

Control of opportunistic yeast is one of the factors reducing nosocomial infection in hospital. In this study, we investigated the presence of virulence factors including antifungal susceptibility, haemolytic potency, proteinase and phospholipase production. In addition biofilm formation was assessed using the XTT assay. A total of 24 Candida strains were isolated and identified. The most common species were Candida albicans and Candida glabrata (40.7%), followed by Candida dubliniensis (8.3%), Candida kefyr (4.2%) and Candida krusei (4.2%). In addition, 62.5% of strains were resistant to itraconazole and 16.7% of strains were resistant to fluconazole. All the strains were susceptible to amphotericin B. Phospholipase activity showed that 8.3% of the total strains were very strong producers and 41.7% were strong producers. Furthermore, three strains (12.5%) were found to have very strong proteinase activity. All the tested strains revealed beta haemolytic potency. All Candida strains were adhesive to polystyrene 96-well microtitre plates to differing degrees. The OD values of XTT reduction at 492 nm ranged from 0.025 (ca12) to 0.927 (ca2). The majority of C. albicans strains were highly metabolically active in comparison with the other species. Control of fungal infections in hospitalised patients is important to prevent the emergence of multi-resistant strains to commonly used antifungal agents.

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References

  • Aoki S, Ito-Kuwa S, Nakamura Y, Masuhara T (1990) Comparative pathogenicity of a wild-type strain and respiratory mutants of Candida albicans in mice. Zentralbl Bakteriol 273:332–343

    CAS  PubMed  Google Scholar 

  • Belanger M, Begin C, Jacques M (1995) Lipopolysaccharides of Actinobacillus pleuropneumoniae bind pig hemoglobin. Infect Immun 63:656–662

    CAS  PubMed  Google Scholar 

  • Bosco VL, Birman EG, Cury AE, Paula CR (2003) Yeasts from the oral cavity of children with AIDS: exoenzyme production and antifungal resistance. Pesqui Odontol Bras 17:217–222

    Article  PubMed  Google Scholar 

  • Branchini ML, Pfaller MA, Rhine-Chalberg J, Frempong T, Isenberg HD (1994) Genotypic variation and slime production among blood and catheter isolates of Candida parapsilosis. J Clin Microbiol 32:452–456

    CAS  PubMed  Google Scholar 

  • Cassone A, De Bernardis F, Mondello F, Ceddia T, Agatensi L (1987) Evidence for a correlation between proteinase secretion and vulvovaginal candidosis. J Infect Dis 156:777–783

    CAS  PubMed  Google Scholar 

  • Chandra J, Mukherjee PK, Leidich SD, Faddoul FF, Hoyer LL, Douglas LJ, Ghannoum MA (2001) Antifungal resistance of candidal biofilms formed on denture acrylic in vitro. J Dent Res 80:903–908

    Article  CAS  PubMed  Google Scholar 

  • Costa CR, Jesuino RS, de Aquino Lemos J, de Fatima Lisboa Fernandes O, Hasimoto ESLK, Passos XS, do Rosario Rodrigues Silva M (2010) Effects of antifungal agents in sap activity of Candida albicans Isolates. Mycopathologia 169:91–98

    Article  PubMed  Google Scholar 

  • Cutler JE (1991) Putative virulence factors of Candida albicans. Annu Rev Microbiol 45:187–218

    Article  CAS  PubMed  Google Scholar 

  • de Oliveira EE, Silva SC, Soares AJ, Attux C, Cruvinel B, Silva Mdo R (1998) Killer toxin and enzyme production by Candida albicans isolated from buccal mucosa in patients with cancer. Rev Soc Bras Med Trop 31:523–527

    Article  PubMed  Google Scholar 

  • Edmond MB, Wallace SE, McClish DK, Pfaller MA, Jones RN, Wenzel RP (1999) Nosocomial bloodstream infections in United States hospitals: a three-year analysis. Clin Infect Dis 29:239–244

    Article  CAS  PubMed  Google Scholar 

  • Flanagan PG, Barnes RA (1998) Fungal infection in the intensive care unit. J Hosp Infect 38:163–177

    Article  CAS  PubMed  Google Scholar 

  • Ghannoum MA (2000) Potential role of phospholipases in virulence and fungal pathogenesis. Clin Microbiol Rev 13:122–143

    Article  CAS  PubMed  Google Scholar 

  • Gristina AG, Giridhar G, Gabriel BL, Naylor PT, Myrvik QN (1993) Cell biology and molecular mechanisms in artificial device infections. Int J Artif Organs 16:755–763

    CAS  PubMed  Google Scholar 

  • Henriques M, Azeredo J, Oliveira R (2007) The involvement of physico-chemical interactions in the adhesion of Candida albicans and Candida dubliniensis to epithelial cells. Mycoses 50:391–396

    Article  PubMed  Google Scholar 

  • Ibrahim AS, Filler SG, Alcouloumre MS, Kozel TR, Edwards JE Jr, Ghannoum MA (1995) Adherence to and damage of endothelial cells by Cryptococcus neoformans in vitro: role of the capsule. Infect Immun 63:4368–4374

    CAS  PubMed  Google Scholar 

  • Jahn B, Martin E, Stueben A, Bhakdi S (1995) Susceptibility testing of Candida albicans and Aspergillus species by a simple microtiter menadione-augmented 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide assay. J Clin Microbiol 33:661–667

    CAS  PubMed  Google Scholar 

  • Jin Y, Samaranayake LP, Samaranayake Y, Yip HK (2004) Biofilm formation of Candida albicans is variably affected by saliva and dietary sugars. Arch Oral Biol 49:789–798

    Article  CAS  PubMed  Google Scholar 

  • Kantarcioglu AS, Yucel A (2002) Phospholipase and protease activities in clinical Candida isolates with reference to the sources of strains. Mycoses 45:160–165

    Article  CAS  PubMed  Google Scholar 

  • Krcmery V, Barnes AJ (2002) Non-albicans Candida spp. causing fungaemia: pathogenicity and antifungal resistance. J Hosp Infect 50:243–260

    Article  CAS  PubMed  Google Scholar 

  • Kuhn DM, Chandra J, Mukherjee PK, Ghannoum MA (2002) Comparison of biofilms formed by Candida albicans and Candida parapsilosis on bioprosthetic surfaces. Infect Immun 70:878–888

    Article  CAS  PubMed  Google Scholar 

  • Kwon-Chung KJ, Lehman D, Good C, Magee PT (1985) Genetic evidence for role of extracellular proteinase in virulence of Candida albicans. Infect Immun 49:571–575

    CAS  PubMed  Google Scholar 

  • Luo G, Samaranayake LP, Yau JY (2001) Candida species exhibit differential in vitro haemolytic activities. J Clin Microbiol 39:2971–2974

    Article  CAS  PubMed  Google Scholar 

  • Manns JM, Mosser DM, Buckley HR (1994) Production of a haemolytic factor by Candida albicans. Infect Immun 62:5154–5156

    CAS  PubMed  Google Scholar 

  • Marr KA, Seidel K, Slavin MA, Bowden RA, Schoch HG, Flowers ME, Corey L, Boeckh M (2000) Prolonged fluconazole prophylaxis is associated with persistent protection against candidiasis-related death in allogeneic marrow transplant recipients: long-term follow-up of a randomized, placebo-controlled trial. Blood 96:2055–2061

    CAS  PubMed  Google Scholar 

  • Melo AS, Padovan AC, Serafim RC, Puzer L, Carmona AK, Juliano NL, Brunstein A, Briones MR (2006) The Candida albicans AAA ATPase homologue of Saccharomyces cerevisiae Rix7p (YLL034c) is essential for proper morphology, biofilm formation and activity of secreted aspartyl proteinases. Genet Mol Res 5:664–687

    CAS  PubMed  Google Scholar 

  • Muller FM, Weig M, Peter J, Walsh TJ (2000) Azole cross-resistance to ketoconazole, fluconazole, itraconazole and voriconazole in clinical Candida albicans isolates from HIV-infected children with oropharyngeal candidosis. J Antimicrob Chemother 46:338–340

    Article  CAS  PubMed  Google Scholar 

  • Neugnot V, Moulin G, Dubreucq E, Bigey F (2002) The lipase/acyltransferase from Candida parapsilosis: molecular cloning and characterization of purified recombinant enzymes. Eur J Biochem 269:1734–1745

    Article  CAS  PubMed  Google Scholar 

  • Panagoda GJ, Ellepola AN, Samaranayake LP (2001) Adhesion of Candida parapsilosis to epithelial and acrylic surfaces correlates with cell surface hydrophobicity. Mycoses 44:29–35

    Article  CAS  PubMed  Google Scholar 

  • Price MF, Wilkinson ID, Gentry LO (1982) Plate method for detection of phospholipase activity in Candida albicans. Sabouraudia 20:7–14

    CAS  PubMed  Google Scholar 

  • Quindos G, Salesa R, Carrillo-Munoz AJ, Lipperheide V, Jaudenes L, San MR, Torres-Rodriguez JM, Ponton J (1994) Multicenter evaluation of ATB fungus: a standardized micromethod for yeast susceptibility testing. Chemotherapy 40:245–251

    Article  CAS  PubMed  Google Scholar 

  • Ramage G, Martinez JP, Lopez-Ribot JL (2006) Candida biofilms on implanted biomaterials: a clinically significant problem. FEMS Yeast Res 6:979–986

    Article  CAS  PubMed  Google Scholar 

  • Rodero L, Cuenca-Estrella M, Cordoba S, Cahn P, Davel G, Kaufman S, Guelfand L, Rodriguez-Tudela JL (2002) Transient fungemia caused by an amphotericin B-resistant isolate of Candida haemulonii. J Clin Microbiol 40:2266–2269

    Article  CAS  PubMed  Google Scholar 

  • Roehm NW, Rodgers GH, Hatfield SM, Glasebrook AL (1991) An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT. J Immunol Methods 142:257–265

    Article  CAS  PubMed  Google Scholar 

  • Safdar A, van Rhee F, Henslee-Downey JP, Singhal S, Mehta J (2001) Candida glabrata and Candida krusei fungemia after high-risk allogeneic marrow transplantation: no adverse effect of low-dose fluconazole prophylaxis on incidence and outcome. Bone Marrow Transplant 28:873–878

    Article  CAS  PubMed  Google Scholar 

  • Segal BH, Bow EJ, Menichetti F (2002) Fungal infections in nontransplant patients with hematologic malignancies. Infect Dis Clin North Am 16:935–964

    Article  PubMed  Google Scholar 

  • Shin JH, Kee SJ, Shin MG, Kim SH, Shin DH, Lee SK, Suh SP, Ryang DW (2002) Biofilm production by isolates of Candida species recovered from nonneutropenic patients: comparison of bloodstream isolates with isolates from other sources. J Clin Microbiol 40:1244–1248

    Article  PubMed  Google Scholar 

  • Waltimo TM, Orstavik D, Meurman JH, Samaranayake LP, Haapasalo MP (2000) In vitro susceptibility of Candida albicans isolates from apical and marginal periodontitis to common antifungal agents. Oral Microbiol Immunol 15:245–248

    Article  CAS  PubMed  Google Scholar 

  • Wingard JR, Niehaus CS, Peterson DE, Jones RJ, Piantadosi S, Levin LS, Saral R, Santos GW (1991) Oral mucositis after bone marrow transplantation. A marker of treatment toxicity and predictor of hepatic veno-occlusive disease. Oral Surg Oral Med Oral Pathol 72:419–424

    Article  CAS  PubMed  Google Scholar 

  • Zink S, Nass T, Rosen P, Ernst JF (1996) Migration of the fungal pathogen Candida albicans across endothelial monolayers. Infect Immun 64:5085–5091

    CAS  PubMed  Google Scholar 

Download references

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Authors and Affiliations

  1. Laboratoire de Bactériologie, Hôpital Régionale de Kairouan, Kairouan, Tunisie

    Kamel Chaieb, Jamel Eddouzi, Yousra Souiden & Kacem Mahdouani

  2. Laboratoire d’Analyse, Traitement, Valorisation des Polluants de l’Environnement et des Produits, Faculté de Pharmacie, Monastir, Tunisie

    Kamel Chaieb & Amina Bakhrouf

Authors
  1. Kamel Chaieb
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  2. Jamel Eddouzi
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  3. Yousra Souiden
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  4. Amina Bakhrouf
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  5. Kacem Mahdouani
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Correspondence to Kamel Chaieb.

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Kamel Chaieb and Jamel Eddouzi contributed equally to this manuscript.

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Chaieb, K., Eddouzi, J., Souiden, Y. et al. Biofilm formation and virulence properties of Candida spp. isolated from hospitalised patients in Tunisia. Ann Microbiol 60, 481–488 (2010). https://doi.org/10.1007/s13213-010-0066-8

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  • DOI: https://doi.org/10.1007/s13213-010-0066-8

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