Candida yeasts are common in the oral cavity and can cause candidosis in the presence of predisposing factors, especially diabetes. The manifestation of the disease is related to this set of local factors such as the presence of dental prostheses, salivary pH, salivary flow and tobacco and the ability to form biofilms. Biofilms are specific and organized communities of cells under the control of signaling molecules rather than random accumulations of cells resulting from cell division and frequently are drugs resistance. Aim: The objectives of this study were to determine the genetic patterns of these C. albicans isolates and to evaluate the in vitro activity amphotericin B and caspofungin against C. albicans biofilms. Methods: Microbial samples were collected from subgingival sites and seeded in CHROMagar for subsequent identification of C. albicans by PCR. Genotypes were defined based on the identification of the transposable introns in the 25S rDNA by PCR. Results: In this study, 6 strains were identified as C. albicans and of these, 3 strains were genotype A and 3 were genotype B. The results showed that both amphotericin B and caspofungin exhibited strong antifungal activities against C. albicans biofilm formation and inhibiting the biofilm formation ranging from 70.8 – 95.3% and 77.7 - 88.7%, respectively. The antifungals studied had low inhibitory effect on preformed biofims, ranging from 39.5 - 50.8% for amphotericin B and from 23.1 - 36.9% for caspofungin at the same concentration. The activity of the two drugs was most effective in inhibit biofilm formation.
This study contributes in existent in literature, demonstrating that clinical isolates that deserve special attention in connection with the difficulty of treatment.
P. Eggimann, J. Garbino, and D. Pittet, "Epidemiology of Candida species infections in critically ill non-immunosuppressed patients," Lancet Infect Dis., vol. 3, pp. 685-702, 2003.
L. C. Shao, C. Q. Sheng, and W. N. Zhang, "Recent advances in the study of antifungal lead compounds with new chemical scaffolds," Yao Xue Xue Bao., vol. 42, pp. 1129-1136, 2007.
CLSI, "Reference method for broth dilution antifungal susceptibility testing of filamentous fungi; approved standard-second edition. CLSI document M38-A2," Clinica and Laboratory Standards Institute (formely NCCLS), 940 West Valley Road, Wayne, Pa, 2008.
J. Sardi, C. Duque, J. Höfling, and R. Gonçalves, "Genetic and phenotypic evaluation of Candida albicans strains isolated from subgingival biofilm of diabetic patients with chronic periodontitis," Med Mycol., vol. 50, pp. 467-75, 2012.
J. Chandra, D. Kuhn, P. Mukherjee, L. Hoyer, T. McCormick, and M. Ghannoum, "Biofilm formation by the fungal pathogen Candida albicans: Development, architecture, and drug resistance," J Bacteriol. Sep., vol. 183, pp. 5385-94, 2001.
F. R?zicka, V. Holá, M. Votava, and R. Tejkalová, "Importance of biofilm in Candida parapsilosis and evaluation of its susceptibility to antifungal agents by colorimetric method," Folia Microbiol (Praha), vol. 52, pp. 209-14, 2007.
E. Nascimento, R. Martinez, and A. Lopes, "Detection and selection of microsatellites in the genome of paracoccidioides brasiliensis as molecular markers for clinical and epidemiological studies," J Clin Microbiol., vol. 42, pp. 5007-5014, 2004.
J. Sardi, A. Fusco-Almeida, and M. Mendes-Giannini, "New antimicrobial therapies used against fungi present in subgingival sites – A brief review," Arch Oral Biol., vol. 56, pp. 951-9, 2011.
M. McCullough, K. Clemons, and D. Stevens, "Molecular and phenotypic characterization of genotypic Candida albicans subgroups and comparison with Candida dubliniensis and Candida stellatoidea," J Clin Microbiol., vol. 37, pp. 417-421, 1999.
C. Pierce, P. Uppuluri, A. Tristan, F. J. Wormley, E. Mowat, G. Ramage, and J. Lopez-Ribot, "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, pp. 1494-500, 2008.
G. Ramage, E. Mowat, B. Jones, C. Williams, and J. Lopez-Ribot, "Our current understanding of fungal biofilms," Crit Rev Microbiol., vol. 35, pp. 340-55, 2009.
S. Tobudic, C. Kratzer, A. Lassnigg, W. Graninger, and E. Presterl, "In vitro activity of antifungal combinations against Candida albicans biofilms," J Antimicrob Chemother., Feb). Epub 2009 Dec 8. PubMed PMID: 19996142, vol. 65, pp. 271-4, 2010.
R. Rajendran, D. P. Robertson, P. J. Hodge, D. F. Lappin, and G. Ramage, "Hydrolytic enzyme production is associated with Candida albicans biofilm formation from patients with type 1 diabetes," Mycopathologia, vol. 170, pp. 229-235, 2010.
D. M. Kuhn and M. A. Ghannoum, "Candida biofilms: Antifungal resistance and emerging therapeutic options," Curr Opin Investig Drugs, vol. 5, pp. 186-197, 2004.
R. Rautemaa and G. Ramage, "Oral candidosis--clinical challenges of a biofilm disease," Crit Rev Microbiol., vol. 37, pp. 328-36, 2011.
P. Gilbert, D. G. Allison, and A. J. McBain, "Biofilms in vitro and in Vivo: Do singular mechanisms imply cross-resistance?," Symp Ser Soc Appl Microbiol., vol. 31, pp. 98-110, 2002.
E. Cateau, M. Rodier, and C. Imbert, "In vitro efficacies of caspofungin or micafungin catheter lock solutions on Candida albicans biofilm growth," J Antimicrob Chemother, vol. 62, pp. 153-155, 2008.
J. Shuford, K. Piper, J. Steckelberg, and R. Patel, "In vitro biofilm characterization and activity of antifungal agents alone and in combination against sessile and planktonic clinical Candida albicans isolates," Diagn Microbiol Infect Dis., vol. 57, pp. 277-281, 2007.
J. Shuford, M. Rouse, K. Piper, J. Steckelberg, and R. Patel, "Evaluation of caspofungin and amphotericin B deoxycholate against Candida albicans biofilms in an experimental intravascular catheter infection model," J Infect Dis., vol. 194, pp. 710- 713, 2006.
Q. Guillermo, y. María Villar-Vidal, and E. Elena, "Actividad de la micafungina contra las biopelículas de Candida," Rev Iberoam Micol., vol. 26, pp. 49-55, 2009.
N. Jain, R. Kohli, E. Cook, P. Gialanella, T. Chang, and B. Fries, "Biofilm formation by and antifungal susceptibility of Candida isolates from urine," Appl Environ Microbiol., vol. 73, pp. 1697-1703, 2007.