315 USE OF PERIFOLLICULAR BLOOD FLOW TO PREDICT THE DEVELOPMENTAL COMPETENCE OF BOVINE CUMULUS–OOCYTE COMPLEXES COLLECTED DURING REPEATED OVUM PICKUP SESSIONS ONCE OR TWICE WEEKLY

Abstract

On average, only 20% of the cumulus–oocyte complexes (COC) develop to the blastocyst stage (Merton et al. 2003 Theriogenology 59, 651–674). An increase in the blood supply to individual follicles appears to be associated with follicular growth rates, whereas a reduction seems to be closely related to follicular atresia (Acosta et al. 2003 Reproduction 125, 759–767). The purpose of this study was to determine whether qualitative perifollicular blood flow changes can be used to predict the developmental competence of COC collected during repeated ovum pickup (OPU) sessions once or twice weekly. Lactating Holstein cows (n = 20) were used as oocyte donors. After dominant follicle removal, OPU was performed twice (group 1, for 3 weeks) or once (group 2, for six weeks) weekly employing a 7.5-MHz transducer (GE 8C-RS) of an ultrasound scanner (GE Logiq Book). Follicle size and Doppler characteristics were recorded by transvaginal ultrasonography just before COC collection using color flow imaging. Owing for technical limitations for measurement of blood flow in small individual follicles, only the presence or absence of blood flow was assessed for each follicle. When a clearly visible blue or red spot (blood flow) was detected in the follicle wall, it was considered as a follicle with detectable blood flow. Follicles with or without detectable blood flow from each individual cow were aspirated separately. After morphological classification of COC, standard protocols for IVP were used for embryo production (Wrenzycki et al. 2001 Biol. Reprod. 65, 323–331). Cleavage and blastocyst rates were recorded at Day 3 and Day 8, respectively. In total, 464 (246 with and 218 without detectable blood flow) and 243 (125 with and 118 without detectable blood flow) follicles ≥3 mm were aspirated in group 1 and group 2, respectively. Morphology of the COC was similar in all groups. Developmental rates for COC stemming from follicles with or without detectable blood flow in group 1 did not show differences for cleavage rates, 54.0% (34/63) and 56.7% (45/81), and for blastocyst rates, 25.4% (16/63) and 22.2% (18/83), respectively. In group 2, the cleavage rates were also similar for COC originating from follicles with and without detectable blood flow, 54.3% (25/46) and 51.5% (34/66). However, developmental rates up to the blastocyst stage did show a significant difference, 23.9% (11/46) and 15.2% (10/66) for COC aspirated from follicles with or without detectable blood flow (P ≤ 0.05). These results show that using COC originating from follicles with detectable perifollicular blood flow collected once weekly may have a higher developmental competence compared to those from follicle without detectable blood flow. Within the detection limits of this study, differences in perifollicular blood flow during repeated OPU sessions once weekly were predictive of oocyte competence.

Ruthe Research Farm, Germany, for providing the animals; Masterrind GmbH, Germany, for donation of the semen; and the HW Schaumann Stiftung for financial support.