Tetrazolium reduction allows assessment of biofilm formation by Campylobacter jejuni in a food matrix model
Article first published online: 20 AUG 2013
© 2013 Crown Copyright. Journal of Applied Microbiology © 2013 Society for Applied Microbiology. This article is published with the permission of the Controller of HMSO and the Queen's Printer for Scotland.
Journal of Applied Microbiology
Volume 115, Issue 5, pages 1212–1221, November 2013
How to Cite
Brown, H.L., van Vliet, A.H.M., Betts, R.P. and Reuter, M. (2013), Tetrazolium reduction allows assessment of biofilm formation by Campylobacter jejuni in a food matrix model. Journal of Applied Microbiology, 115: 1212–1221. doi: 10.1111/jam.12316
- Issue published online: 15 OCT 2013
- Article first published online: 20 AUG 2013
- Accepted manuscript online: 1 AUG 2013 01:55AM EST
- Manuscript Accepted: 25 JUL 2013
- Manuscript Revised: 28 JUN 2013
- Manuscript Received: 5 APR 2013
- Biotechnology and Biological Sciences Research Council (BBSRC
- BBSRC Institute Strategic Program. Grant Number: IFR/08/3 and BB/J004529/1
- BBSRC CASE studentship. Grant Number: BB/I15321/1
- 2,3,5 triphenyltetrazolium chloride;
- Campylobacter jejuni ;
- crystal violet;
- food matrix
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.
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.