| This experiment collected 180 ovaries from 48 yellow catties, 34 buffalos and 8 holstein catties at the slaughter house, and acquired 1050 oocytes. There were 973 oocytes used among them. Aspiration of 68 buffalo ovaries had provided an average of 3.22 utilizable oocytes per ovary, which was about 1/2 obtained from yellow cows or holstein catties ovaries(6.72 and 6.81 respectively), there was significant difference between them (P<0.01). Ratio (87.50%) of recovery buffalo oocytes applied to IVM was significantly lower than that of catties and Holstein catties oocytes(94.59% and 96.69% respectively, P<0.01). The results indicate that growing follicles of buffalo ovaries are little, the yield of oocytes is little and quantity is bad.This research compared the methods of three kinds of ovum collection. Recovery oocytes with the method of aspiration and incision together were an average of 8.39 per ovary, and recovery oocytes used were 7.71, significantly higher than those oocytes with the method of aspiration (6.31 and 6.19, respectively, P<0.01) and method of incision (6.97 and 6.44, respectively, PO.01); Recovery oocytes per ovary by incision were significantly higher than those by aspiration(P<0.05), but recovery oocytes used were no significantly difference from those by aspiration (P>0.05) .The results indicate that recovery naked oocytes with the method of incision were high. The method of aspiration and incision together is a good method of collecting more oocytes, but its procedure is complicated, it is suitable for collecting a few ovaries once.Without 17 3 -E2 in culture media for IVM of follicular oocytes, adding 0.3% PVP and 10% FBS respectively, the maturation rate of cattle oocytes is 60.69% and 63.46%respectively, the fertilization rate of cattle oocytes is 51.93% and 53.95% respectively, there was no significant difference between them(P>0.05); With 17 P -E2 in culture media, adding 0.3% PVP and 10% FBS respectively, the maturation rate of cattle oocytes is 77.71% and 80.48% respectively, the fertilization rate of cattle oocytes is 64.47% and 67.68% respectively, there was also no significant difference between them (P>0.05) . The results indicate that 0.3% PVP substituting for 10% FBS is feasible in culture media. The same conclusion was reached in this experiment with buffalo oocytes. In addition, adding 17 P -? in culture media for IVM of follicular oocytes of cattle, Holstein cattle and buffalo, the maturation rate and fertilization rate in vitro of oocytes were both significantly higher than those of without adding 17 P -E2( PO.01 or P<0.05). The result shows 17 P -E2 is indispensable to the maturation and fertilization in vitro of bovine oocytes. At the same time, the aim of this study was to elucidate the possibility of applying bovine IVF procedure on buffalo. The result shows that although buffalo oocytes can mature and fertilize in vitro culture, the maturation rate and fertilization rate of buffalo oocytes were significantly lower than those of cattle oocytes under identical conditions ( P<0.05 or PO.01 ), this shows applying bovine IVF procedure on buffalo is available, but possibly fertilization media or certain steps for using in this procedure does not really suit buffalo oocytes, still needs the further improvement. From now on the research point in buffalo IVF should be direct towards advancing culture effectiveness for IVM, in order to improve the entire efficiency on buffalo IVF.This experiment studied two kinds of different methods to separate frozen-thawed bovine spermatozoa before fertilization. After separation spermatozoa by swim-up and Percoll gradient, the fertilization rate of Holstein cattle oocytes was 61.75% and 59.76% respectively. There was no significantly difference between them (P>0.05) , the same conclusion was obtained on study of buffalo and cattle. The result shows swim-up or Percoll method is used to separate spermatozoa, but the Percoll prodcedure is more simple, faster than the swim-up method.Therefore, it is the first choice method, if we can a...
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