American Journal of Infectious Diseases and Microbiology
ISSN (Print): 2328-4056 ISSN (Online): 2328-4064 Website: Editor-in-chief: Maysaa El Sayed Zaki
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American Journal of Infectious Diseases and Microbiology. 2018, 6(2), 62-65
DOI: 10.12691/ajidm-6-2-5
Open AccessArticle

Optimising a Fusarium solani Biofilm Formation Protocol in Vitro

Marwan Y. Al-Maqtoofi1,

1Department of Biology, College of Sciences, University of Basrah, 61004, Iraq

Pub. Date: October 16, 2018

Cite this paper:
Marwan Y. Al-Maqtoofi. Optimising a Fusarium solani Biofilm Formation Protocol in Vitro. American Journal of Infectious Diseases and Microbiology. 2018; 6(2):62-65. doi: 10.12691/ajidm-6-2-5


Opportunistic fungi belonging to the Fusarium solani have become increasingly recognised as life-threatening pathogens causing keratitis and disseminated fusariosis among both healthy individuals and patients with haematological malignancies. These infections are associated with biofilm formation on different biotic and abiotic surfaces. Considering, a biofilm is a virulence factor for causing infections, the aim of this study optimising and illustrating a simple, cost-effective and highly reproducible 96 well microtitre-based method for F. solani biofilm formation via using crystal violet stain. The results revealed that the possibility of using either 570nm or 595nm as a wavelength for quantifying fungal biofilm formation. The best time for crystal violet de-staining was 10 min of incubation. This model can be used in-vitro to quantify and understand the virulence factor of fungal biofilm during infections, and for antifungal susceptibility testing.

Biofilm formation Fusarium solani fungi crystal violet 96 well microtitre plates

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[1]  Al-Maqtoofi M, Thornton CR. Detection of human pathogenic Fusarium species in hospital and communal sink biofilms by using a highly specific monoclonal antibody, Environ Microbiol, vol. 18, no. 11, pp. 3620-3634, Nov. 2016.
[2]  Guarro J. Fusariosis, a complex infection caused by a high diversity of fungal species refractory to treatment, Eur. J. Clin. Microbiol. Infect. Dis., vol. 32, no. 12, pp. 1491-1500, Dec. 2013.
[3]  Kaur S, Singh, S. Biofilm formation by Aspergillus fumigatus, Med Mycol, vol. 52, no. 1, pp. 2-9, Jan. 2014.
[4]  Nett JE, Cain MT, Crawford K, Andes DR. Optimizing a Candida Biofilm Microtiter Plate Model for Measurement of Antifungal Susceptibility by Tetrazolium Salt Assay, J. Clin. Microbiol., vol. 49, no. 4, pp. 1426-1433, Apr. 2011.
[5]  Zago CE, Silva S, Sanitá PV, Barbugli PA, Dias CM, Lordello VB, Vergani CE. Dynamics of Biofilm Formation and the Interaction between Candida albicans and Methicillin-Susceptible (MSSA) and -Resistant Staphylococcus aureus (MRSA), PLoS One, vol. 10, no. 4, Apr. 2015.
[6]  Taff HT, Nett JE, Andes DR. Comparative analysis of Candida biofilm quantitation assays, Med Mycol, vol. 50, no. 2, pp. 214-218, Feb. 2012.
[7]  Mowat E, Butcher J, Lang S, Williams C, Ramage G. Development of a simple model for studying the effects of antifungal agents on multicellular communities of Aspergillus fumigatus, J. Med. Microbiol., vol. 56, no. Pt 9, pp. 1205-1212, Sep. 2007.
[8]  Pathak AK, Sharma S, Shrivastva P. Multi-species biofilm of Candida albicans and non-Candida albicans Candida species on acrylic substrate’, Journal of Applied Oral Science, vol. 20, no. 1, pp. 70-75, Feb. 2012.
[9]  Pierce CG, Uppuluri P, Tristan AR, Wormley FL, Mowat E, Ramage G, Lopez-Ribot JL. A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing, Nat Protoc, vol. 3, no. 9, pp. 1494-1500, 2008.
[10]  Silva-Dias A, Miranda IM, Branco J, Monteiro-Soares M, Pina-Vaz C, Rodrigues AG. Adhesion, biofilm formation, cell surface hydrophobicity, and antifungal planktonic susceptibility: relationship among Candida spp., Front. Microbiol., vol. 6, 2015.
[11]  Melo AS, Bizerra FC, Freymüller E, Arthington-Skaggs BA, Colombo AL. Biofilm production and evaluation of antifungal susceptibility amongst clinical Candida spp. isolates, including strains of the Candida parapsilosis complex, Med Mycol, vol. 49, no. 3, pp. 253-262, Apr. 2011.
[12]  Seidler MJ, Salvenmoser S, Müller MC. Aspergillus fumigatus Forms Biofilms with Reduced Antifungal Drug Susceptibility on Bronchial Epithelial Cells, Antimicrob. Agents Chemother., vol. 52, no. 11, pp. 4130-4136, Nov. 2008.
[13]  Chandra J, Kuhn DM, Mukherjee PK, Hoyer LL, McCormick T, Ghannoum MA, Biofilm Formation by the Fungal Pathogen Candida albicans: Development, Architecture, and Drug Resistance, J. Bacteriol., vol. 183, no. 18, pp. 5385-5394, Sep. 2001.
[14]  Peiqian L, Xiaoming P, Huifang S, Jingxin Z, Ning H, Birun L. Biofilm formation by Fusarium oxysporum f. sp. cucumerinum and susceptibility to environmental stress, FEMS Microbiol. Lett., vol. 350, no. 2, pp. 138-145, Jan. 2014.
[15]  Ramage G, Rajendran R, Gutierrez-Correa M, Jones B, Williams C. Aspergillus biofilms: clinical and industrial significance, FEMS Microbiol. Lett., vol. 324, no. 2, pp. 89–97, Nov. 2011.
[16]  Imamura Y, Chandra J, Mukherjee PK, Lattif AA, Szczotka-Flynn LB, Pearlman E, Jonathan HL, O’Donnell K, Ghannoum MA. Fusarium and Candida albicans Biofilms on Soft Contact Lenses: Model Development, Influence of Lens Type, and Susceptibility to Lens Care Solutions, Antimicrob Agents Chemother, vol. 52, no. 1, pp. 171-182, Jan. 2008.
[17]  Li X, Yan Z, Xu J. ‘Quantitative variation of biofilms among strains in natural populations of Candida albicans’, Microbiology (Reading, Engl.), vol. 149, no. Pt 2, pp. 353-362, Feb. 2003.
[18]  Pitts B, Hamilton MA, Zelver N, Stewart PS. A microtiter-plate screening method for biofilm disinfection and removal’, J. Microbiol. Methods, vol. 54, no. 2, pp. 269-276, Aug. 2003.