title: Cytotoxicity of four trichothecenes evaluated by three colorimetric bioassays.
abstract: The application of cell culture technique for screening of low concentrations of Fusarium mycotoxins was examined. Three colorimetric bioassays were used to determine the cytotoxicity of the trichothecenes T-2 toxin (T-2), HT-2 toxin (HT-2), deoxynivalenol (DON) and nivalenol (NIV) to 3T3 mouse fibroblasts (3T3 cells). The bioassays assess DNA synthesis (incorporation of 5-bromo-2'-deoxyuridine; BrdU), metabolic activity (cleavage of 3-(4,5-dimethyltiazol-2-yl)-2,5-diphenyltetrazolium bromide; MTT) and cell membrane damage (release of lactate dehydrogenase; LDH), respectively. The BrdU bioassay was the most sensitive and the IC50 values (50% response compared to untreated cells) of T-2, HT-2, DON and NIV were 4.6, 13, 263 and 365 ng/ml, respectively. At the same toxin concentrations used in the BrdU bioassay, only T-2 and HT-2 were toxic enough to obtain IC50 values using the MTT bioassay. The IC50 values for T-2 and HT-2 were 12 and 68 ng/ml, respectively. When determined by the LDH bioassay, the IC50 values of T-2 and HT-2 were 18 and 42 ng/ml, respectively. At the tested concentrations, DON and NIV had a minor effect on the 3T3 cells when evaluated by the MTT and LDH bioassays. The BrdU bioassay in combination with 3T3 cells was found to be a suitable method for determination of trichothecene-induced toxicity at low concentrations.
authors.affiliation: Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences, Uppsala, Sweden. Johan.Widestrand@huv.slu.se
authors.forename: J
authors.initials: J
authors.lastname: Widestrand
authors.forename: T
authors.initials: T
authors.lastname: Lundh
authors.forename: H
authors.initials: H
authors.lastname: Pettersson
authors.forename: J E
authors.initials: JE
authors.lastname: Lindberg
mesh_terms: D016475:3T3 Cells / Q000187:drug effects*; D000818:Animals; D001973:Bromodeoxyuridine; D003124:Colorimetry; D004305:Dose-Response Relationship, Drug; D007770:L-Lactate Dehydrogenase; D051379:Mice; D013605:T-2 Toxin / Q000031:analogs & derivatives / Q000633:toxicity; D013778:Tetrazolium Salts; D018675:Toxicity Tests / Q000379:methods; D014255:Trichothecenes / Q000633:toxicity*
chemical_list: D013778:Tetrazolium Salts; D014255:Trichothecenes; C038405:nivalenol; D007770:L-Lactate Dehydrogenase; D001973:Bromodeoxyuridine; D013605:T-2 Toxin; C007262:deoxynivalenol; C012351:HT-2 toxin