The role of honeybee pollen as a natural source of antioxidants in the in vitro maturation medium of sheep oocytes and its effect on gene expression.

This study aimed to investigate the effect of honeybee pollen as an antioxidant source in a maturation medium of sheep oocytes on the in vitro maturation rate, glutathione concentration, and gene expression. To our knowledge, this study might be the first of its kind in this field. Sheep oocytes were cultured in vitro with honeybee pollen at four different concentrations (0.0, 1.0, 10.0, and 50.0 µg/ml). The results indicated that the ratio of oocytes that reached metaphase II stage was higher in the honeybee pollen-treated groups than in the control group (p ≤ 0.05). The reduced glutathione (GSH) mean content of matured oocytes was 9.85 nmol/25 oocytes, when honeybee pollen was added to the in vitro maturation (IVM) medium at a concentration of 1.0 µg/ml, compared with 5.84 and 4.44 nmol when using 10.0 and 50.0 µg/ml honeybee pollen, respectively. On the other hand, there was no significant difference in glutathione concentration between the control and 1.0 µg/ml honeybee pollen groups. Expression of candidate genes (GDF-9, BAX, Cyclin B, C-MOS, and IGF1) was upregulated in oocytes cultured with honeybee pollen when compared with oocytes cultured without honeybee pollen. In conclusion, the addition of honeybee pollen at a concentration of 1.0 µg/ml to IVM medium improved the in vitro maturation rate of sheep oocytes, increased the glutathione concentration, and improved gene expression.

Use of black seed (Nigella sativa) honey bee to improve sheep oocyte maturation medium.

Sheep are important livestock and a source of milk, meat, and wool globally. The increasing demand for animal protein requires increased productivity in sheep. In vitro fertilization and maturation can improve sheep productivity. The aims of this study were to evaluate the effects of honey bee addition as a supplementation medium on in vitro maturation improvement, gene expression of matured sheep oocytes, and determine the optimum concentration from honey bee for in vitro maturation of sheep oocytes. Cumulus oocyte complexes were obtained from the ovaries of slaughtered female sheep. Grade A and B oocytes were cultured for 24 h in medium without honey bee (control, G1) or medium supplemented with 5% (G2), 10% (G3), or 20% (G4) honey bee. Oocyte maturation rate, glutathione concentration, and the expression of candidate genes (GDF-9, BAX, Cyclin B, C-MOS, IGF1) were determined in the matured oocytes. The maturation rate of sheep oocyte was better in the presence of 5% and 10% honey bee; the mean number of oocytes in metaphase II stage was higher than that in G1 and G4 groups. Glutathione concentration was highest in G2 (10.93 ± 0.57). In general, gene expression levels were similar in G2 and G3, which were greater that in G1 and G4. In conclusion, the optimal concentration of black seeds honey bee that can be added to the maturation medium is 5% to obtain the highest mean MII and glutathione concentration values, and to improve gene expression in in vitro matured sheep oocytes.