A rapid and sensitive cytotoxicity screening assay for trichothecenes in cereal samples.
The application of a bioassay for toxicity screening of cereal samples has been studied. The sensitive Swiss mouse 3T3 fibroblasts (3T3 cells) were used to determine the cytotoxicity of Fusarium trichothecenes T-2 toxin (T-2), HT-2 toxin (HT-2), deoxynivalenol (DON) and nivalenol (NIV) in cereal samples. Clean-up of the sample extracts was performed using the MycoSep #225 column and the cytotoxic effects of the samples were determined using the 5-bromo-2'-deoxyuridine (BrdU) bioassay assessing DNA-synthesis. Trichothecene-free samples of wheat, wheat bran and barley had no toxic effect on the 3T3 cells at sample concentrations up to 400 mg/ml cell culture medium while a few oat samples had an inhibiting effect on the DNA-synthesis. IC(50) values (50% response compared with untreated cells) of trichothecene-free wheat and oats spiked with T-2, HT-2, DON and NIV were similar to IC(50) values of pure toxins. A cytotoxicity screening of 33 different cereal samples showed that all samples contaminated with a trichothecene concentration higher than the IC(50) values of T-2, HT-2, DON and NIV had a clear inhibiting effect on DNA-synthesis of the 3T3 cells. The culture of 3T3 cells on microtiter plates in combination with the simple and rapid sample clean-up using the MycoSep #225 column provides a sensitive and easy method for the screening of trichothecenes in cereal samples.
Widestrand
J
J
Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences, S-75007 Uppsala, Sweden.
Lundh
T
T
Pettersson
H
H
Lindberg
J E
JE
D016475:3T3 Cells; D000818:Animals; D000963:Antimetabolites; D018554:Avena / Q000737:chemistry; D001681:Biological Assay; D001973:Bromodeoxyuridine; D002470:Cell Survival / Q000187:drug effects*; D002523:Edible Grain / Q000737:chemistry*; D001467:Hordeum / Q000737:chemistry; D007202:Indicators and Reagents; D051379:Mice; D009183:Mycotoxins / Q000633:toxicity; D019384:Nucleic Acid Synthesis Inhibitors / Q000633:toxicity*; D011500:Protein Synthesis Inhibitors / Q000633:toxicity*; D014255:Trichothecenes / Q000633:toxicity*; D014908:Triticum / Q000737:chemistry
D000963:Antimetabolites; D007202:Indicators and Reagents; D009183:Mycotoxins; D019384:Nucleic Acid Synthesis Inhibitors; D011500:Protein Synthesis Inhibitors; D014255:Trichothecenes; D001973:Bromodeoxyuridine