The study was conducted to determine the effects of holding water buffalo and cattle ovaries during transport and storage at various temperatures on in vitro developmental capacity of the retrieved oocytes. Ovaries were collected from slaughterhouses and held in saline solution at temperatures of 37, 32, and 15ºC during transport to laboratory and further storage. Aspiration and culture of oocytes were done at 4-6 or 16-18 hr post collection. In experiment 1, oocytes from ovaries stored at different temperatures were matured in vitro for 22-24 hr and were investigated for nuclear maturation based on 1st polar body extrusion. In experiment 2, oocytes were matured, fertilized in vitro and evaluated for cleavage and blastocyst development. Results indicate that oocytes from ovaries held for 4- 6 hr at 37, 32 and 15ºC did not differ significantly in terms of rates of maturation, cleavage and development to blastocyst in both species. When ovaries were stored for 16-18 hr, the rates of maturation and subsequent development in 32 and 37ºC groups were significantly lower than those of 15ºC group. The results suggest that prolonged storage of ovaries at high temperatures decreases the developmental competence of oocytes, while storage of ovaries at low temperature (15ºC) does not affect in vitro production of embryos in both species. The present study demonstrated for the first time the success of storing water buffalo ovaries at low temperature (15ºC) for longer period which can maximize the utilization of valuable germplasm for animal production with huge potential application in animal biodiversity conservation.