Post-warming hatching and birth of live calves following transfer of in vitro-derived vitrified water buffalo (Bubalus bubalis) embryos

Abstract

Viability of in vitro-derived vitrified-warmed preimplantation stage buffalo embryos were assessed in vitro and in vivo. Oocytes were collected from ovaries of slaughtered riverine buffaloes, matured and fertilized in vitro with frozen semen from riverine buffalo bull and cultured on cumulus cell monolayers. Resultant preimplantation stage embryos were cryopreserved by vitrification with ethylene glycol, ficoll and sucrose. Seventy-one frozen embryos were warmed in 0.5M sucrose and were further cultured in vitro for 72 h to assess hatching rate. On the other hand, 95 embryos were transferred non-surgically to riverine buffalo recipients to assess development competence in vivo through detection of pregnancy and birth of live calves. Hatching rate of 83.10% (59/71) was noted among embryos cultured in vitro. Pregnancy rate was 16.36% (9/55) while calving rate was 10.91% (6/55) after transfer of in vitro-derived vitrified-warmed embryos to recipient animals. Six healthy and normal calves with average birth weight of 38.75+/-3.55 kg were born from the transferred embryos. These results indicate the viability of vitrified in vitro-derived buffalo embryos and the potential application of in vitro embryo production and vitrification techniques for production and transport of buffalo embryos from germplasm-rich sources to guarantee genetic improvement in many parts of the world.