Adaptation of an [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay to evaluate the cytotoxicity of the extracellular products of micro-organisms pathogenic to fish.
OBJECTIVE
Adaptation of a colorimetric assay using [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] (MTT) to evaluate the cytotoxicity of the extracellular products of micro-organisms pathogenic to fish.


RESULTS
The optimal conditions for the colorimetric assay were determined and this method was compared with the trypan blue exclusion assay. The protein concentration of extracellular products causing the death of 50% of the cell population (CI50) was determined.


CONCLUSIONS
This assay enables quantitative and objective comparison of the cytotoxicity of the extracellular products of micro-organisms pathogenic to fish. It was shown to be more accurate than conventional counting with the trypan blue exclusion assay.


CONCLUSIONS
This method may also be useful for characterizing the cytotoxicity of specific components of extracellular products.
Zorrilla
I
I
Department of Microbiology, Faculty of Sciences, University of Malaga, Campus de Teatinos, 29071-Malaga, Spain.
Balebona
M C
MC
Moriñigo
M A
MA
D000818:Animals; D001419:Bacteria / Q000472:pathogenicity*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000494:pharmacology*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003124:Colorimetry; D021541:Sea Bream / Q000382:microbiology*; D013778:Tetrazolium Salts / Q000378:metabolism*; D013844:Thiazoles / Q000378:metabolism*; D014343:Trypan Blue
D001426:Bacterial Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; C022616:thiazolyl blue; D014343:Trypan Blue