Peroxisome proliferator-activated receptor delta-PPARδ agonist (L-165041) enhances bovine embryo survival and post vitrification viability.

The effect of L-165041 (PPARδ-agonist) on decreasing apoptosis and intracellular lipid content was assessed in fresh and vitrified-warmed in vitro -produced bovine embryos. It was hypothesised that the addition of L-165041 to the culture medium enhances development and cryopreservation. Oocytes were allocated to one of two treatments: control-standard culture medium, or L-165041 added to the medium on day1 with no media change. Ultrastructure, cleavage, and blastocyst rates were evaluated in fresh, and in post-vitrification cultured embryos by optical and electronic microscopy. A subset of fresh embryos were fixed for TUNEL assay and for Sudan-Black-B histochemical staining. Vitrified-warmed embryos were assessed using MALDI-MS technique. Cleavage and blastocyst rates (control 49.4±5.2, L-165041 51.8±4.3) were not influenced by L-165041. The proportion of inner cell mass cells (ICM) was higher in fresh embryos, and the rate of total and ICM apoptosis was lower in L-165041. In warmed-embryos, total and ICM apoptosis was lower in L-165041. The overall hatching rate was higher in L-165041 (66.62±2.83% vs 53.19±2.90%). There was less lipid accumulation in fresh L-165041-embryos. In conclusion, the use of L-165041 is recommended to improve the viability of in vitro -derived bovine embryos.

Antral follicular count and its relationship with ovarian volume, preantral follicle population and survival, oocyte meiotic progression and ultrastructure of in vitro matured bovine cumulus-oocyte complexes.

This study aimed to evaluate the relationship between antral follicular count (AFC) and ovarian volume (OV), preantral follicular population and survival, meiotic progression and ultrastructure of cumulus-oocyte complexes (COCs) after in vitro maturation. In experiment 1, the relationship between AFC and preantral follicle population and survival was evaluated by classical histology. In experiment 2, the relationship among AFC, OV, ability of oocytes to resume meiosis and ultrastructure of in vitro matured bovine COCs was studied. A positive correlation (P < 0.05) between AFC and the numbers of healthy primordial, degenerate and total follicles was observed, as well as with healthy secondary follicles and total follicles. The numbers of grades I and II oocytes in ovaries of high AFC class were higher compared with those with intermediate or lower AFC. After in vitro maturation, COCs from ovaries of high AFC had a higher percentage of oocytes in metaphase II compared with those of intermediate and low AFC (P < 0.0001). Ovaries of intermediate AFC had a higher percentage of oocytes in metaphase II compared with ovaries with low AFC (P < 0.0001). The proportion of oocytes in metaphase I, telophase I and anaphase I in COCs from ovaries of intermediate AFC (26.04%) was higher (P < 0.05) compared with that seen in COCs of ovaries with high (8.55%) and low (14.15%) AFC. No differences in the ultrastructure of oocytes were seen. In conclusion, after in vitro maturation, cow ovaries with high AFC have higher numbers of oocytes that reach in metaphase II (MII), but they also have higher numbers of degenerated primordial and primary follicles.

Effects of vulvar width and antral follicle count on oocyte quality, in vitro embryo production and pregnancy rate in Bos taurus taurus and Bos taurus indicus cows.

It was hypothesized that Brahman cows have greater in vitro embryo production than Simmental cows, and that regardless of breed, antral follicle count (AFC) and vulvar width (VW) are positively associated with oocyte quality, embryo number and pregnancy rate. Brahman (Bos taurus indicus, n = 184) and Simmental (Bos taurus taurus, n = 99) cows were classified as having a large AFC (Brahman: ≥50, Simmental: ≥25); intermediate AFC (Brahman: >30 and ≤49; Simmental: >16 and <24) and small AFC (Brahman: <30, Simmental: ≤15). The VW was considered to be large (Brahman: >123 mm, Simmental: >80 mm); intermediate (Brahman: >102 mm and ≤123 mm; Simmental: >65 mm and ≤80 mm) and small (Brahman: ≤102 mm and Simmental: ≤65 mm). For each group of cows, ovum pick up, oocyte in vitro maturation, fertilization and embryo culture were performed. Embryo transfers were subsequently performed and pregnancy rate was evaluated. Brahman cows had a larger number of viable oocytes, greater efficiency of embryo production and greater pregnancy rate (P < 0.05) than Simmental cows. Pregnancy percentages in Simmental cows with a small AFC were greater than those cows with an intermediate and large AFC. With the Brahman cows, there was no difference in pregnancy rate among the cows with different AFC classifications. In contrast to Simmental cows, Brahman cows with an intermediate VW had a greater pregnancy percentage than those with a small and large AFC. In conclusion, Brahman cows have a larger AFC, number of viable oocytes, and pregnancy rate than Simmental cows.