Inherent inferior quality of follicular fluid in repeat breeder heifers as evidenced by low rates of in vitro production of bovine embryos

- Bovine in vitro oocyte maturation, fertilization and blastocyst yield are adversely affected using follicular fluid of the ovulatory follicle from repeat breeder dairy heifers.
- Our findings suggest the existence of an inherent inferior quality of the ovulatory follicle microenvironment in repeat breeding Holstein heifers.

The aims of the present study were to determine the effect of follicular fluid obtained from the ovulatory follicle of repeat breeder heifers on in vitro oocyte maturation (Experiment 1), fertilization (Experiment 2) and production of bovine embryos (Experiment 3). Holstein virgin heifers (VH, n = 5) with normal fertility or repeat breeder syndrome (RBH, n = 5) were used in the present study. Follicular fluid of VH and RBH was aspirated from ovulatory follicles and used as maturation medium. Bovine oocytes were aspirated from follicles of slaughterhouse ovaries and randomly allocated in three groups; in Group 1, oocytes cultured in TCM-199 supplemented with 10% heat-treated fetal calf serum and hormones (5 IU/mL hCG plus 0.1 IU/mL rFSH); in Group 2, oocytes cultured in TCM-199 supplemented with 10% filtered follicular fluid of VH without hormones; in Group 3, oocytes cultured in TCM-199 supplemented with 10% filtered follicular fluid of RBH without hormones. The mean (±SEM) percentage of matured oocytes was different between VH and RBH groups (72.2 ± 4.0 vs 56.4 ± 4.6%; P < 0.05, respectively). Further, the mean (±SEM) percentage of normal oocyte fertilization was higher in the VH than the RBH group (49.3 ± 2.1 vs 32.0 ± 4.2; P < 0.05, respectively). The mean percentage of embryos developed to the blastocyst stage was higher in the VH than the RBH group (12.0 ± 1.3 vs 7.0 ± 1.6; respectively; P < 0.05). In conclusion, our findings support our hypothesis that the ovulatory follicle microenvironment of Holstein repeat breeder heifers places their oocytes at a developmental disadvantage compared with Holstein fertile virgin heifers and that this suggest the existence of an inherent inferior quality of the ovulatory follicle microenvironment in repeat breeding Holstein heifers.