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.

Fatty acid profiles of the plasma and follicular fluid mares fed a combination of linseed and salmon oil.

This study evaluated the presence of polyunsaturated fatty acids in circulating blood and in the ovarian follicular fluid of mares, after supplementation of the diet with linseed oil. Six Mangalarga Marchador mares, weighing 397.00±31.89 kg, were kept on native pasture, and assigned to the current study. In a switch over design, mares were randomly allocated to receive 150 ml of vegetable oil daily, containing polyunsaturated fatty acids n3 (62.23 g ALA, 20.34 g LA, 2.27 g EPA, 2.32 g DHA), (n=3) or no supplementation (n=3) in two replicates. Blood and follicular fluid samples were taken on the first day (D0) and every 30 days until the end of the supplementation period (D60). After 60 days of supplementation, mares were switched across the treatments. Plasma concentrations of linolenic acid in total fatty acids were higher (P=0.006) in the supplemented compared to the control group (1.89±0.13 vs. 1.49±0.13%). There were positive correlations between plasma linoleic acid and follicular fluid arachidonic acid (P=0.0106; r2=0.13) and between plasma alpha linolenic acid and follicular fluid EPA (P=0.0004; r2=0.2544). Data indicated a low to moderate relationship between the dietary linseed-based oil supplementation studied and circulating and follicular fluid polyunsaturated fatty acids contents in mares.

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.

Anti-Müllerian Hormone (AMH), antral follicle count (AFC), external morphometrics and fertility in Tabapuã cows.

The intense selection of characteristics related to animal performance may affect the reproductive efficiency of beef cows negatively. Selection for fertility along with production traits is, therefore, readily justifiable. The objective of the present study was to evaluate possible relationships among AFC, serum AMH concentrations and external morphometrics with reproductive efficiency in Tabapuã (a Brazilian Bos indicus beef breed) females. Antral follicle counts and external morphometrics were measured in nulliparous (n = 162), primiparous (n = 80) and multiparous (n = 351) cows, from four farms. Age at first parturition (AFP), parturition interval (PI), maternal ability (MA), precocity, rusticity and survival (PRS) and overall reproductive efficiency (RE) were evaluated according to the Brazilian Association of Zebu Breeders (ABCZ) data bank indexes. A single blood sample per animal was collected at random stages of the reproductive cycle from a subset of animals (nulliparous = 84, primiparous and multiparous = 136) to determine serum AMH concentrations. The AFC classes were defined as lesser (<28), intermediate (28-50) and greater (>50). Correlations between all variables and the effects of parity and AFC on reproductive traits, AMH and external genitalia size were analyzed by the PROCORR and by the PROCGENMOD procedures of SAS® (SAS, Cary, NC, USA), respectively. Antral follicle count did not differ (P = 0.71) among nulliparous (38.6 ±â€¯23.96) primiparous (47.54 ±â€¯26.16) and multiparous (41.08 ±â€¯25) cows and was negatively correlated with pregnancy interval (PI), such that, as PI decreased (r = -0.28; P < 0.005), AFC increased. Vulva width was not affected (P = 0.08) by parity and was larger (P < 0.05) for females in the greater AFC class (8.81 ±â€¯0.12 cm) compared with the intermediate (8.42 ±â€¯0.11 cm) and lesser (8.38 ±â€¯0.13 cm) classes. As vulva width increased, parturition interval decreased (r = -0.15; P < 0.005) and overall reproductive efficiency increased (r = 0.17, p < 0.005). Thoracic depth was associated with greater AFCs (r = 0.10; P < 0.001). Anti-Müllerian hormone concentrations were greater (P < 0.05) for animals in the greater AFC class (1.15 ±â€¯0.09 ng/mL) compared with the lesser (0.44 ±â€¯0.02 ng/mL) and intermediate (0.73 ±â€¯0.05 ng/mL) classes. Vulva width, AFC, external morphometrics and AMH concentrations were moderately associated, considering the low heritability of fertility traits, and should be studied further to be considered in the selection for fertility in Bos indicus cattle.

Vulvar width and rima length as predictors of the ovarian follicular reserve in bovine females.

The aim of this study was to investigate possible relationships between morphometric characteristics of the bovine reproductive tract and measures of fertility, such as antral follicle counts (AFCs) and the number and quality of recovered cumulus oophorus complexes (COCs). First, the genital tracts of 360 abattoir Zebu/Holstein crossbred cows were studied. Rima vulvae length (RL), vulvar width (VW), ovarian size, pelvic fat, number of aspirated COCs, and AFCs were recorded. An index of COC quality (CQI) was established, which weighed overall gamete quality based on the IETS classification. Second, the same external measurements and AFCs (by ultrasound) were analyzed in live Tabapuã (Zebu) cows (n = 48). Relative RL and vulvar width were defined as the original measurements divided by the body weight of each cow for statistical analyses. In abattoir tracts, the AFC was smaller (P < 0.05) in animals with large VW (34.88 ± 3.50) than in animals with small (45.71 ± 2.57) and intermediate (42.25 ± 2.45) VW. The CQI was higher (P < 0.05) when the pelvic fat score was intermediate (3.22 ± 0.06) than that in lean (2.99 ± 0.08) and obese (2.90 ± 0.12) animals. The CQI was higher (P < 0.05) in the intermediate ovary group (3.19 ± 0.08) than in the small (2.96 ± 0.08) and large (2.95 ± 0.09) ovary groups. In live cows, the AFC was higher (P = 0.035) in females with large relative rima length than in those with small relative rima length (18.96 ± 1.97 and 14.76 ± 1.51, respectively). Similarly, AFC was greater (P = 0.0001) in females with large relative VW than in those with small relative VW (20.08 ± 1.36 and 11.16 ± 1.60, respectively). In conclusion, larger external genitalia relative to body size were good predictors of the ovarian follicular reserve in live animals.