Effect of alpha-tocopherol and ascorbic acid on bovine in vitro fertilization

The aim of this work was to study the effect of alpha-tocopherol (vitamin E) and ascorbic acid on the in vitro fertilization process. Frozen bovine semen was prepared using extenders with and without addition of vitamin E. Samples were capacitated with heparin in the fertilization medium. In vitro matured oocytes were inseminated with spermatozoa frozen with and without vitamin E and, after thawing, fertilized in TALP medium (control) and in TALP medium with vitamin E (1 mg/ml), with ascorbic acid (5 mM) and with vitamin E plus ascorbic acid. Gametes were incubated in the respective fertilization medium for 48 h; those frozen without vitamin E yielded 75, 76, 69 and 49% of fertilized oocytes in the control, vitamin E, ascorbic acid and vitamin E plus ascorbic acid media, respectively. The last value was significantly different (P < 0.01). In bovine sperm frozen with vitamin E, fertilization rates were 74, 50, 47 and 34%, respectively for the 4 groups. Values observed for the different supplements were significantly different inter se (P < 0.01), except between the media with vitamin E and with ascorbic acid. These results indicate that preserved antioxidant capacity of vitamin E impairs the success of the in vitro fertilization process.