Collection of oocytes through transvaginal ultrasound-guided aspiration of follicles in an Indian breed of cattle

摘要

The present study was undertaken in Karan Fries, an Indian breed of cattle to (1) determine the number of follicles available for puncture and (2) explore the potential of this breed as a donor of developmentally competent oocytes. Ovum pick-up (OPU) was performed using an ultrasound machine with a transvaginal convex transducer (5 MHz) with a needle guide, single lumen 19-gauge 60 cm long needle and a vacuum pressure of 90 mmHg. The number and size of follicles in each ovary was determined before puncture. The follicles were characterized on the basis of their diameter as small (3-5 mm), medium (6-9 mm) and large (greater than or equal to10 mm). The oocytes recovered were classified by quality. They were matured in vitro, irrespective of their grade, in 50 mul droplets of the in vitro maturation (IVM) medium (TCM-199 + 10% fetal bovine serum (FBS) + 5 mug/ml follicle stimulating hormone (folltropin) + 1 mug/ml estradiol-17beta+0.2 mM sodium pyruvate), covered with paraffin oil, in 35 mm petridish for 24 h in a CO2 incubator (5% CO2 in air) at 38.5degreesC. The cleavage rate was recorded at day 2 post-insemination after subjecting the oocytes to in vitro fertilization (IVF). The differences in follicular populations of all size categories among individual donors were not significant. A total of 92 oocytes were recovered by aspiration of 157 follicles, with an overall recovery rate of 59% (range 35-79%). Of these, 32% were of grades A and B and the rest of grades C and D. The mean numbers of total follicles and the oocytes recovered per session did not differ significantly among individual donors. Out of the 73 oocytes subjected to IVM and IVF, 24 reached 2-4 cell stage at day 2 post-fertilization, with a cleavage rate of 33%. The total number of oocytes recovered was correlated with the number of small (R = 0.54, P < 0.01) but not with the number of medium and large follicles. This study demonstrates the use of OPU as a means of obtaining developmentally competent oocytes from an Indian breed of cattle for obtaining cattle oocytes in India where cow slaughter is not allowed for religious reasons.