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Rapid colorimetric bioassay for screening of Fusarium mycotoxins
B A Rotter1, B K Thompson, S Clarkin
1Centre for Food and Animal Research, Agriculture, Canada, Ottawa, Ontario.
Abstract:
The cytotoxicity of Fusarium mycotoxins was evaluated using a trichothecene sensitive cell line (BHK-21, baby hamster kidney cells) in combination with the MTT-cleavage test as an end-point measurement. Cells tended to be more sensitive to the type A trichothecenes with midpoint cytotoxicity values ranging from 1.6 ng/ml for T-2 toxin to 60 ng/ml for scirpentriol. The cytotoxicity value for deoxynivalenol (type B) was 112 ng/ml. The inherent disadvantage of the MTT-assay (formation of insoluble formazan) was overcome by using the analog MTS and measuring the water-soluble formazan directly in the culture media. The MTS-midpoint cytotoxicity values for T-2 toxin and deoxynivalenol (2.1 and 141 ng/ml, respectively), although slightly higher, showed a good correspondence to the MTT-test. Both the MTT- and MTS-cleavage tests are useful for evaluating the cytotoxicity of Fusarium mycotoxins. The replacement of MTT by MTS substantially reduced the number of sample processing steps and the length of time required to complete the cytotoxicity assay.
Insights
This study assessed Fusarium mycotoxin cytotoxicity using baby hamster kidney cells. The MTS assay offers a faster, simpler method for evaluating mycotoxin toxicity compared to the MTT assay.
Area of Science:
- Toxicology
- Cell Biology
- Biochemistry
Background:
- Fusarium mycotoxins pose a significant threat to human and animal health.
- Accurate assessment of mycotoxin cytotoxicity is crucial for risk evaluation.
- Traditional assays like MTT have limitations in terms of processing and time.
Purpose of the Study:
- To evaluate the cytotoxicity of Fusarium mycotoxins using a trichothecene-sensitive cell line.
- To compare the efficacy of the MTT-cleavage assay with its analog, the MTS assay.
- To determine the suitability of the MTS assay for rapid and efficient mycotoxin cytotoxicity testing.
Main Methods:
- Utilized a baby hamster kidney (BHK-21) cell line, sensitive to trichothecenes.
- Employed the MTT-cleavage assay to measure cytotoxicity.
- Implemented the MTS assay, which measures water-soluble formazan directly in culture media, overcoming MTT's limitations.
Main Results:
- Type A trichothecenes, such as T-2 toxin (1.6 ng/ml) and scirpentriol (60 ng/ml), exhibited higher cytotoxicity than Type B deoxynivalenol (112 ng/ml) using the MTT assay.
- MTS assay yielded comparable cytotoxicity values for T-2 toxin (2.1 ng/ml) and deoxynivalenol (141 ng/ml), showing good correlation with MTT results.
- The MTS assay significantly reduced sample processing steps and assay time.
Conclusions:
- Both MTT and MTS assays are effective for evaluating Fusarium mycotoxin cytotoxicity.
- The MTS assay provides a more streamlined and time-efficient alternative to the MTT assay.
- The findings support the use of the MTS assay for routine and high-throughput screening of mycotoxin toxicity.