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Type: Artigo de periódico
Title: Proposal of protocols using D-glutamine to optimize the 2,3-bis (2-methoxy-4-nitro-5-sulfophenly)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) assay for indirect estimation of microbial loads in biofilms of medical importance
Author: Gobor, T
Corol, G
Ferreira, LEN
Rymovicz, AUM
Rosa, RT
Campelo, PMS
Rosa, EAR
Abstract: Due to technical problems, biofilm biomasses are difficult to be precisely determined. One reliable strategy is based on the colorimetry of formazan compounds derived from tetrazolium salt reduction. XTT presents some desirable properties that make the biofilm measurements easier. However, cells entrapped within the extracellular matrixes normally do not metabolize the tetrazolium equally, leading to underestimation of cell contents. This study evaluated the effectiveness of D-glutamine, a plerotic substrate of tricarboxilic acid cycle (TAC), as inducer of KIT reduction. The metabolic activities of aerobic and anaerobic 48 h-old monospecific biofilms of Pseudomonas aeruginosa ATCC (R) 27853 (TM). Klebsiella pneumoniae ATCC (R) 13883 (TM). Staphylococcus epidermidis ATCC (R) 12228 (TM). Streptococcus mutans ATCC (R) 25175 (TM), and Candida albicans SC5314 were evaluated. Results showed that D-glutamine 50 mM (for P. aeruginosa, K. pneumoniae, and S. epidermidis) and 25 mM (for S. mutans and C albicans) may enhance the detection of soluble formazan in a significant manner, what becomes the XTT reduction assay more robust. (C) 2011 Elsevier B.V. All rights reserved.
Subject: Biofilms
XTT reduction assay
Metabolic activity
Country: Holanda
Editor: Elsevier Science Bv
Rights: fechado
Identifier DOI: 10.1016/j.mimet.2010.12.018
Date Issue: 2011
Appears in Collections:Artigos e Materiais de Revistas Científicas - Unicamp

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