Improved 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay for measuring the viability of lactic acid bacteria

Abstract

The aim of this study was to adapt MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] colorimetric assay (MCA) (Wang et al., 2010) for determination of lactic acid bacteria cell count. Our study is helpful in developing protocols for measuring the viability of other lactic acid bacteria. We determined the cell concentration range which gives linear regression of CFU (colony forming units) and formazan production. In our experiment three authentic lactic acid bacteria strains were investigated: Lactobacillus rhamnosus VT1, L. plantarum 2142, and L. sakei DSM 2001. As data show beside the strain also the growth medium has influence on the MTT reduction activity, this means that for each special case separate calibration curve has to be prepared. Based on MTT reduction we also developed an improved microtiter plate assay which proved to be reliable for a rapid cell count determination. So our methods are only applicable for estimate cell number with fixed parameters, but under given circumstances they are fast and sensitive methods. With the modified methods we can rapidly measure the dehydrogenase enzyme activity of the lactic acid bacteria cells. With the microplate assay we can measure many conditions and strains at the same time.