Assessment of biofilms formation of bacterial and fungal isolates using qualitative Congo red agar and semiquantitative crystal violet microtiter methods

Xiomara Moreno , Melanie Ventura, María Mercedes Panizo, María Fátima Garcés, .

Keywords: Biofilms, Gram-positive bacteria, Gram-negative bacteria, yeasts, Congo red

Abstract

Introduction. Sixty-five percent of human infections are caused by bacteria or yeasts able to form biofilms. This feature makes them more resistant to antimicrobials and antifungals.
Objective. To determine biofilm formation capacity of bacterial and fungal isolates by quantitative crystal violet microtiter and qualitative Congo red agar methods.
Materials and methods. Brain-heart infusion, trypticase soy broth and Müeller‑Hinton culture media were used in bacterial isolates for the quantitative method; brain-heart infusion broth and Sabouraud dextrose were used for yeasts. The same culture media plus 3% Congo red and 10% dextrose were used to apply the qualitative method in agar. The proposal by Stepanovic, et al. was used as a reference method.
Results. We evaluated 103 bacterial isolates and 108 yeasts isolates. We did not recommend substitute brain-heart infusion broth for trypticase soy and Müeller-Hinton broths for biofilm formation assessment in bacterial isolates using the quantitative method. Sabouraud dextrose medium, both broth and agar, can replace brain-heart infusion to assess biofilm formation in yeasts, quantitatively and qualitatively.
Conclusion. The study of biofilms in the microbiology laboratory, using Congo red agar qualitative method, is a simple, fast, and inexpensive procedure that provides precise information for the diagnosis and treatment of persistent infections caused by bacteria and yeasts.

Downloads

Download data is not yet available.

References

Cheong JZA, Johnson CJ, Wan H, Liu A, Kernien JF, Gibson ALF, et al. Priority effects dictate community structure and alter virulence of fungal-bacterial biofilms. ISME J. 2021;15:2012-27. https://doi.org/10.1038/s41396-021-00901-5

Ibarra C, Villar M, Gaitán L, Pozos A, Mendoza R, Sánchez L. Ensayo de formación y cuantificación de biopelículas mixtas de Candida albicans y Staphylococcus aureus. Rev Iberoam Micol. 2012;29:214-22. https://doi.org/10.1016/j.riam.2012.02.003

Moreno X, Armas A, Panizo M, Garcés M. Formación de biopelículas por Klebsiella pneumoniae aislada en heces de pacientes con trastorno del espectro autista. Rev Soc Ven Microbiol. 2020;40:12-8. https://doi.org/10.5281/zenodo.3660110

Moreno X, Marcano E, Panizo M, Ferrara G. Formación de biopelículas por Candida parapsilosis sensu stricto y su perfil de susceptibilidad en Venezuela. Kasmera. 2020;48:1-7. https://doi.org/10.5281/zenodo.3660110

Ortega S, Hernández E. Biopelículas microbianas y su impacto en áreas médicas: fisiopatología, diagnóstico y tratamiento. Bol Med Hosp Infant Mex. 2018;75:79‑88. https://doi.org/10.24875/BMHIM.M18000012

Díaz A, Vivas R, Puerta L, Ahumedo M, Arévalo L, Cabrales R, et al. Biopelículas como expresión del mecanismo de quorum sensing: una revisión. Av Periodon Implantol. 2011;23:195-201.

Murray P, Rosenthal K, Pfaller M. Microbiología médica. 7th Edition. Washington, D.C.: ASM Press; 2013.

Eraso M. Principales características de las biopelículas relacionadas con procesos patológicos descritos en humanos en los últimos 10 años, revisión sistemática. Investig Andina. 2016;18:1491-506. https://doi.org/10.33132/01248146.556

Gómez J, Gómez M, Bas P, Ramos C, Cafini F, Maestre J, et al. ¿Es la cuantificación del biofilm un elemento diferenciador en la patogenia de bacilos gramnegativos? Rev Esp Quimioter. 2013;26:97-102.

Stepanović S, Vuković D, Dakić I, Savić B, Švabić-Vlahović M. A modified microtiter-plate test for quantification of staphylococcal biofilm formation. J Microbiol Met. 2000;40:175-9. https://doi.org/10.1016/S0167-7012(00)00122-6

Kragh KN, Alhede M, Kvich L, Bjarnsholt T. Into the well -A close look at the complex structures of a microtiter biofilm and the crystal violet assay. Biofilm. 2019;1:100006. https://doi.org/10.1016/j.bioflm.2019.100006

Rampelotto R, Lorenzoni V, Silva C, Coelho S, Wust V, Garzon L, et al. Assessment of different methods for the detection of biofilm production in coagulase-negative staphylococci isolated from blood cultures of newborns. Rev Soc Bras Med Trop. 2018;51:761-7. https://doi.org/10.1590/0037-8682-0171-2018

Oliveira A, Cunha M. Comparison of methods for the detection of biofilm production in coagulase-negative staphylococci. BMC Res Notes. 2010;3:260. https://doi.org/10.1186/1756-0500-3-260

Hassan A, Usman J, Kaleem F, Omair M, Khalid A, Iqbal M. Evaluation of different detection methods of biofilm formation in the clinical isolates. Braz J Infect Dis. 2011;15:305-11. https://doi.org/10.1016/s1413-8670(11)70197-0

da Costa Lima JL, Alves LR, Pereira da Paz JN, Rabelo MA, Vieira-Maciel MA, Camargo de Morais MM. Analysis of biofilm production by clinical isolates of Pseudomonas aeruginosa from patients with ventilator-associated pneumonia. Rev Bras Ter Inten. 2017;29:310-6. https://doi.org/10.5935/0103-507x.20170039

Achek R, Hotzel H, Nabi I, Kechida S, Mami D, Didouh N, et al. Phenotypic and molecular detection of biofilm formation in Staphylococcus aureus isolated from different sources in Algeria. Pathogens. 2020;9:153. https://doi.org/10.3390/pathogens9020153

Peña J, Uffo O. Producción de biofilm en genotipos de Staphylococcus aureus aislados de mastitis bovina en Cuba. Rev Salud Anim. 2013;35:189-96.

Vuotto C, Longo F, Pascolini C, Donelli G, Balice MP, Libori MF, et al. Biofilm formation and antibiotic resistance in Klebsiella pneumoniae urinary strains. J Appl Microbiol. 2017;123:1003-18. https://doi.org/10.1111/jam.13533

IIknur D, Kiraz N, Oz Y. Evaluation of different detection methods of biofilm formation in clinical Candida isolates. Afr J Microbiol Res. 2010;4:2763-8. https://doi.org/10.5897/AJMR.9000302

Rivera J, Aranaga V, Reyes G, Vega O, Ciancio L, Arencio L, et al. Adherencia y formación de biopelícula sobre superficies abióticas lisas en Staphylococcus spp., aislados de quesos artesanales e industriales. Boletín del Centro de Investigaciones Biológicas de la Universidad del Zulia. 2018;52:38-55.

Mathur T, Singhal S, Khan S, Upadhyay DJ, Fatma T, Rattan A. Detection of biofilm formation among the clinical isolates of staphylococci: An evaluation of three different screening methods. Indian J Med Microbiol. 2006;24:25-9. https://doi.org/10.1016/S0255-0857(21)02466-X

Moreno X, Núñez G, Rosales O, Ferrara G, Panizo MM. Evaluación de factores de virulencia y perfil de susceptibilidad en Candida albicans provenientes de secreciones vaginales. Ac SVBE. 2021;24:70-78221;24.

How to Cite
1.
Moreno X, Ventura M, Panizo MM, Garcés MF. Assessment of biofilms formation of bacterial and fungal isolates using qualitative Congo red agar and semiquantitative crystal violet microtiter methods. biomedica [Internet]. 2022 Aug. 31 [cited 2024 May 13];43(Sp. 1):77-88. Available from: https://revistabiomedica.org/index.php/biomedica/article/view/6732

Some similar items:

Published
2022-08-31

Altmetric

Article metrics
Abstract views
Galley vies
PDF Views
HTML views
Other views
Crossref Cited-by logo
QR Code