Tetrazolium reduction allows assessment of biofilm formation by Campylobacter jejuni in a food matrix model

Brown, Helen, van Vliet, Arnoud, Betts, Roy and Reuter, Mark (2013) Tetrazolium reduction allows assessment of biofilm formation by Campylobacter jejuni in a food matrix model. Journal of Applied Microbiology, 115 (5). pp. 1212-1221. ISSN 1364-5072

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Abstract

AIMS: To develop a staining method for specific detection of metabolically active (viable) cells in biofilms of the foodborne pathogen Campylobacter jejuni. METHODS AND RESULTS: Conversion of 2,3,5 triphenyltetrazolium chloride (TTC) to insoluble, red 1,3,5-triphenylformazan (TPF) was dependent on metabolic activity of Camp. jejuni. When used with chicken juice, TTC staining allowed quantification of Camp. jejuni biofilm levels, whereas the commonly used dye, crystal violet, gave high levels of nonspecific staining of food matrix components (chicken juice). The assay was optimized to allow for monitoring of biofilm levels and adapted to monitor levels of Camp. jejuni in broth media. CONCLUSIONS: Staining with TTC allows for the quantification of metabolically active Camp. jejuni and thus allows for quantification of viable cells in biofilms and food matrices. The TTC staining method can be adapted to quantify bacterial cell concentration in a food matrix model, where the accepted method of A600 measurement is not suitable due to interference by components of the food matrix. SIGNIFICANCE AND IMPACT OF THE STUDY: 2,3,5 Triphenyltetrazolium chloride (TTC) staining is a low-cost technique suitable for use in biofilm analysis, allowing rapid and simple imaging of metabolically active cells and increasing the methods available for biofilm assessment and quantification.

Item Type:	Article
Uncontrolled Keywords:	2,3,5 triphenyltetrazolium chloride,biofilm,campylobacter jejuni ,crystal violet,food matrix
Faculty \ School:	Faculty of Science > School of Biological Sciences Faculty of Medicine and Health Sciences > Norwich Medical School
Depositing User:	Pure Connector
Date Deposited:	30 Oct 2014 14:32
Last Modified:	31 Oct 2022 16:31
URI:	https://ueaeprints.uea.ac.uk/id/eprint/50500
DOI:	10.1111/jam.12316

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