Evaluation of aflatoxin B1 embryotoxicity using the frog embryo teratogenesis assay-Xenopus and bio-activation with microsome activation systems.

BACKGROUND

Aflatoxins are a group of mycotoxins produced by Aspergillus, A. flavus, and A. parasiticus. Aflatoxin B1 (AFB1) should be a strong teratogen in hamsters, but its effect in rats is equivocal and extremely limited in mice. Therefore, the AFB1 embryotoxic potential in mammals remains unclear.

METHODS

Little is known about the AFB1 effects on amphibians, therefore its embryotoxic potential was evaluated using the frog embryo teratogenesis assay-Xenopus (FETAX). X. laevis blastulae were exposed to: 1) positive controls for bio-activation (4 g/L cyclophosphamide monohydrate, Cy, and 4 g/L Cy+30 mg/L MAS-rat; 4 g/L Cy+30 mg/L MAS-human); 2) positive controls for MAS (30 mg/L MAS-rat and 30 mg/L MAS-human); 3) exposed groups to AFB1 (1 mg/L AFB1); and 4) AFB1 bio-activation (1 mg/L AFB1+30 mg/L MAS-rat and 1 mg/L AFB1 +30 mg/L MAS-human).

RESULTS

In MAS-rat and human, Cy did not induce a statistically significant increase of mortality and malformed larvae percentage, but when bio-activated Cy increased the percentage of mortality. Instead, MAS-rat and human alone did not show any increase of mortality and malformed larvae percentages. When bio-activated by MAS-rat and human, AFB1 increased significantly both the mortality and malformed larvae percentages. The malformed larvae were mainly plurimalformed, i.e., affected by generalized edema, abnormal gut coiling, and microphthalmia.

CONCLUSIONS

This research shows that AFB1 alone is not embryotoxic but, when bio-activated with MAS-rat or MAS-human the percentage of mortality and malformed larvae increased significantly. These results also show that AFB1 must be bio-activated to exert its embryotoxic effects.