Superovulating cattle with corifollitropin-alpha, a long-acting recombinant human FSH (rhFSH): Dose-response, half-life, effects on the ovaries, and embryo outcomes.

The aim of this study was to evaluate the superestimulatory and superovulatory responses of cattle treated with corifollitropin-alpha, a long-acting human recombinant FSH (rhFSH). In the first and second experiments, we used Nelore (Bos indicus) heifers previously submitted to follicular wave suppression by active immunization against GnRH. In Experiment 1 (a dose-response study), heifers (n = 20) were randomly allocated into five groups, which received placebo (saline) or a single sc dose of 7.5, 15.0, 22.5 or 30.0 µg rhFSH. The heifers were subjected to daily ovarian scan and blood sampling during 11 days. We observed group, time, and group x time effects (P<0.0001) for both average follicle size and circulating FSH concentrations, with a strong correlation (R = 0.82, P<0.0001) between the area under curve (AUC) for both parameters. The peak concentration of FSH 24h after treatment and average follicle size at all timepoints, however, were similar (P>0.05) between groups 22.5 and 30.0 µg. In Experiment 2, heifers (n = 18) were allocated into three groups, which received (0h) either placebo (control), 25 µg rhFSH or 130 mg pFSH (Folltropin). There was no difference (P>0.05) in average follicle size at any moment, as well as in intrafollicular E2 at 120h or in plasma P4 seven days later between groups rhFSH and pFSH. In Experiment 3, cycling Nelore heifers (n = 20) were subjected to a wave synchronization protocol and superovulated (day 0) using a standard pFSH protocol (120 mg split in eight decreasing im doses) or with a single sc injection of 20 µg rhFSH. The number of follicles >7 mm on day 4 did not differ (P=0.4370). Heifers receiving rhFSH had greater average follicle size on day 4 (P=0.0005), ovulation rate (P<0.0001), and number of CL (P=0.0155), as well as a trend towards a greater number of ova (P=0.07) and viable embryos (P=0.0590). In Experiment 4, superovulation was induced with a single sc injection of 25 µg rhFSH in Girolando and Nelore cows and heifers (n = 20). None of the embryo yield endpoints differed between the two breeds (P>0.05). In conclusion, cattle superstimulation and superovulation can be successfully induced with a single dose of a long-acting rhFSH (corifollitropin-alpha).

Evaluation of the effects of the recommended oral dose of diflubenzuron on bovine sperm and oocyte quality using CASA and OPU-IVEP.

Introduction: The aim of this study was to evaluate potential effects of diflubenzuron on the production and quality of gametes, and on in vitro embryo production (IVEP) outcomes, in cattle. Methods: Two experiments were performed, the first to evaluate effects on semen, and the second on cumulus-oocyte complexes (COC) and on IVEP. Nelore (Bos taurus indicus) bulls (n = 14) or heifers (n = 16) were allocated into control (CG) or treatment (DIF) groups. All groups received a mineral mix supplement added (DIF) or not (CG) with diflubenzuron (30 mg/head/day), during 8 weeks. Animals were weighed and blood samples were collected throughout the experimental period. Every other week, bulls were subjected to semen collection and heifers to transvaginal ultrasound-guided follicle aspiration sessions. Semen underwent physical and morphological evaluation, and samples were stored for further computer-assisted sperm analysis. The COC recovered were evaluated according to morphology and those classified as viable were sent to an IVEP laboratory. Results: Diflubenzuron had no effect (P > 0.05) on average body weight or in any blood hematological or biochemical endpoints, regardless of gender. In experiment 1, there was no difference (P > 0.05) between DIF and CG groups for sperm concentration, morphology, or kinetics. In experiment 2, there was also no effect of diflubenzuron on the number of total, viable, or grade I oocytes, as well as on cleavage or blastocyst rates (P > 0.05). Discussion: In summary, the oral administration of diflubenzuron, within the recommended dose, has no short-term negative effects on sperm production and quality or on oocyte yield and developmental potential in vitro, in cattle.

Dynamics of the Reproductive Changes and Acquisition of Oocyte Competence in Nelore (Bos taurus indicus) Calves during the Early and Intermediate Prepubertal Periods

The purpose of this study was to characterize the reproductive physiology, oocyte competence, and chromatin compaction in Nelore calves in the early-prepubertal period (EPP) and the intermediate-prepubertal period (IPP). Calves aged 2-5 (EPP) and 8-11 months old (IPP) were assigned to Trial 1 (morpho-physiological-endocrine evaluations, n = 8) or Trial 2 (oocyte donors, n = 8) vs. the respective control groups of cows (n = 8, each). All morphological endpoints, except the antral follicle count, increased from the EPP to the IPP. The EPP LH-FSH plasma concentrations were similar to cows, whereas LH was lower and FSH was higher in the IPP than in cows. . Cows produced more Grade I (12.9% vs. 4.1% and 1.7%) and fewer Grade III COC (30.1% vs. 44.5% and 49.0%) than the EPP and IPP calves, respectively. The IPP calves' oocyte diameter was similar to those from cows but greater than those from EPP females (124.8 ± 8.5 and 126.0 ± 7.5 µm vs. 121.3 ± 7.5 µm, respectively). The expression of the chromatin compaction-related gene HDAC3 was downregulated in calves. The proportion of the blastocyst rate to the controls was lower in EPP than in IPP calves (43.7% vs. 78.7%, respectively). Progressive oocyte competence was found during the prepubertal period, which can help to decide whether to recover oocytes from calves.

Hormonal stimulation in 4 to 7 months old Nelore (Bos taurus indicus) females improved ovarian follicular responses but not the in vitro embryo production.

The inclusion of pre-pubertal bovine females in reproductive management could allow in vitro embryo production and reduce generation interval, thereby causing faster genetic gain of the herd. However, oocytes of pre-pubertal females have lower competence, blastocyst production, and pregnancy rates than those collected from pubertal animals. This study aimed to evaluate the effect of an induced hormonal stimulation on the serum concentrations of Anti-Mullerian hormone (AMH) and FSH, ovarian responses, ovum pick up (OPU), and in vitro produced embryos (IVP) from oocytes obtained from four-to seven-months old Nelore female cattle. In a crossover design, these females were randomly allocated into: 1) Treated Group (TG, n = 9): the animals were subjected to a hormonal protocol (implanted progesterone device, estradiol benzoate, LH, and FSH) from Day 0 (the start of the treatment) to Day 7 (OPU day), and 2) Control Group (CG, n = 9): the females did not receive any hormonal stimulation, but they had ablation of their largest follicles on Day 2 of experiment. Blood collection for serum FSH measurements was done on Days 5, 6, 7, and 8, and collection for serum AMH measurements was done on Days 5 and 8. As hypothesized, TG had higher serum FSH concentrations (p < 0.05) on Day 5 (1.16 ±â€¯0.31 ng/mL), Day 6 (1.21 ±â€¯0.45 ng/mL), and Day 7 (0.95 ±â€¯0.26 ng/mL) than CG (0.56 ±â€¯0.17 ng/mL on Day 5, 0.60 ±â€¯0.25 ng/mL on Day 6, and 0.60 ±â€¯0.14 ng/mL on Day 7). However, serum AMH concentrations were neither significantly different (p > 0.05) between CG and TG, nor between the collection days. Hormonal stimulation also increased (p < 0.05) total follicular population (20.0 ±â€¯4.95 CG vs 26.66 ±â€¯4.24 TG), ovarian diameter (13.08 ±â€¯1.0 mm CG vs 14.81 ±â€¯1.38 mm TG) and number of follicles ≥2.5 mm (6.88 ±â€¯2.14 CG vs 11.55 ±â€¯4.09 TG). In TG, grades I and II oocytes predominated, whereas, in CG grades III and IV oocytes were more abundant (p < 0.05). No significant increases (p > 0.05) in the cleavage (49.33% CG vs 51.42% TG), cleavage > 4 cells (9.33% CG vs 16.19% TG), and blastocysts rates (1.33% CG vs 8.57% TG) were seen in TG. This hormonal protocol increased serum FSH concentrations that possibly contributed to increases in the observed follicle, as well as improving oocyte quality. This exogenous hormonal stimulation increased available oocytes numbers for IVP, despite no increase in the in vitro embryo production efficiency.