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Evaluating the toxicity of metabolites derived from the trichothecene biotransformation using Biomin® BBSH 797 in vitro.
Biomin® BBSH 797, a Gen. nov. sp. nov. of the Familie Coriobateriaceae isolated from bovine rumen fluid, is able to specifically detoxify trichothecenes in the intestinal tract of animals. The live strain is able to produce enzymes that reduce the epoxide group of these mycotoxins, which results in the formation of non-toxic metabolites.
The objective of this trial was to prove that the metabolite de-epoxy-deoxynivalenol (DOM-1), formed during the degradation of deoxynivalenol (DON), has lower toxicity.
The lymphocyte proliferation assay (LPA) was used to show the effects of DON and DOM-1 on the clonal proliferation on isolated chicken lymphocytes. Lymphocytes were cultured with DON (10-0.08 µg/mL) and DOM-1 (232-1.18 µg/mL). As a control, the mitogen Concanavaline A (ConA) was used. The number of proliferated cells was quantified with a colorimetric immunoassay after in vitro stimulation. 5-bromo-2’-deoxyuridine (BrdU) was added and incorporated into the DNA of proliferating cells. Incorporated BrdU was measured with an ELISA-method. The absorbance values directly correlated with the amount of DNA synthesis and hence the number of proliferating cells.
Results show that at a concentration of 15 µg DON/mL the proliferation of lymphocytes was lower in comparison to ConA. After adding 0.3 µg DON/mL to the cells only one-third of them could proliferate whereas at a concentration of 0.63 µg DON/mL the growth of the lymphocytes stopped. In comparison, the metabolite DOM-1 only stopped the proliferation of cells at a concentration of 113 µg DOM-1/mL. The results show that the metabolite DOM-1 is almost 500 times less toxic than DON.
Keywords:
Biomin BBSH 797, Trichothecene, Biotransformation