Effects of follicle-stimulating hormone followed by gonadotropin-releasing hormone on embryo production by ovum pick-up and in vitro fertilization in the river buffalo (Bubalus bubalis)

In this study, we examined the effects of superstimulation using follicle-stimulating hormone (FSH) followed by gonadotropin-releasing hormone (GnRH) on buffalo embryo production by ultrasound-guided ovum pick-up (OPU) and in vitro fertilization (IVF). Nine Murrah buffaloes were subjected to OPU-IVF without superstimulation (control). The morphologies of the oocytes collected were evaluated, and oocytes were then submitted to in vitro maturation (IVM). Two days after OPU, same nine buffaloes were treated with twice-daily injections of FSH for 3 days for superstimulation followed by a GnRH injection. Oocytes were collected by OPU 23-24 hr after the GnRH injection and submitted to IVM (the superstimulated group). The total number of follicles, number of follicles with a diameter > 8 mm, and number of oocytes surrounded by multi-layered cumulus cells were higher in the superstimulated group than in the control group (p = 0.05). After IVF, the percentages of cleavage and development to blastocysts were higher in the superstimulated group than in the control group (p < 0.05). In conclusion, superstimulation improved the quality of oocytes and the embryo productivity of OPU-IVF in river buffaloes.