207 Oocyte Recovery Rates and in Vitro Blastocyst Production in Cattle Treated With a Single Injection of Folltropin-v Diluted in a Slow-Release Formulation

Abstract

We have previously shown that superstimulation of beef donors with a single intramuscular (IM) injection of Folltropin-V (Bioniche Animal Health, Belleville, Ontario, Canada) diluted in a proprietary slow-release formulation (SRF; Bioniche Animal Health) results in in vivo embryo production comparable with the traditional twice-daily IM administration over 4 days (Tribulo et al. 2010 Reprod. Fertil. Dev. 22, 367). Two experiments were designed to evaluate oocyte recovery and in vitro blastocyst production of oocyte donors superstimulated with Folltropin-V, administered in twice-daily injections or diluted in SRF and given by a single injection. In Experiment 1, crossbred beef cows (n = 11) and heifers (n = 5) were randomly allocated to 3 treatment groups in a crossover design (i.e. all animals were treated 3 times, and all treatments were represented in the 3 replicates). All donors had all follicles >8 mm in diameter ablated by ultrasound-guided follicle aspiration on Day 0, received 500 μg of cloprostenol (prostaglandin F2α, Ciclase, Syntex, Argentina), and were treated as follows: Group 1 (multiple FSH): 160 mg of Folltropin-V divided into 4 IM injections administered twice daily (i.e. Days 1 and 2); Group 2 (single FSH): 160 mg of Folltropin-V diluted in 4 mL of 25% SRF and given by a single IM injection in the neck on Day 1; Group 3 (control): no FSH treatment. On Day 4, oocytes were aspirated from all follicles >3 mm in diameter by ultrasound-guided follicular aspiration, classified, matured in TCM-199 supplemented with 0.4% BSA, fertilized in Fert medium, and in vitro cultured in SOF supplemented with 0.4% BSA under oil at 37°C, 5% CO2, 5% O2, 90% N2, and saturated humidity for 7 days. Data were analyzed by ANOVA. The mean (±SEM) numbers of follicles aspirated and oocytes recovered were higher (P < 0.05) in Groups 1 (multiple FSH, 13.6 ± 1.9 and 8.5 ± 1.0) and 2 (single FSH, 14.7 ± 1.7 and 9.4 ± 1.3) than in Group 3 (control, 8.7 ± 0.9 and 5.8 ± 1.0). The mean number of blastocysts after 7 days in culture was numerically higher (P = 0.2) in Groups 1 (2.1 ± 0.7) and 2 (2.6 ± 0.6) than in Group 3 (1.1 ± 0.4). In Experiment 2, lactating Holstein cows (n = 10) received 2.5 mg of estradiol benzoate and 50 mg of progesterone IM on Day 0. On Day 4, all cows received prostaglandin F2α and were randomly allocated to 2 treatment groups, to receive 200 mg of Folltropin-V divided into 4 twice-daily IM injections or 200 mg of Folltropin-V diluted in 25% SRF and given as a single IM injection. On Day 7, oocytes were collected and treated like those in Experiment 1. The mean (±SEM) numbers of follicles aspirated (21.2 ± 3.5 and 14.4 ± 2.9), oocytes recovered (10.2 ± 1.2 and 13.8 ± 2.9), and blastocysts produced (4.6 ± 0.7 and 6.0 ± 2.0) did not differ for Groups 1 and 2, respectively (P = 0.3). In conclusion, superstimulatory treatment increased the number of cumulus–oocyte complexes available for IVF compared with no treatment, and ovarian superstimulatory treatment may be simplified by the use of a single IM dose of a sustained-release product, compared with multiple doses of a conventional product, without a reduction in the recovery or quality of cumulus–oocyte complexes.

Efficacy of a single intramuscular injection of porcine FSH in hyaluronan prior to ovum pick-up in Holstein cattle

“…Although a two injection protocol seemed to be required to superovulate donor cows for in vivo embryo production, the results of the present study support the notion that only one injection of pFSH diluted in 0.5% HA is required for in vitro embryo production after OPU in nonlactating Holstein donors. These results are also in agreement with those obtained after a single injection of pFSH diluted in 0.5% HA in beef cows [44]. The present data verify a shortened but extended period of elevated plasma concentrations of FSH in HA groups.…”

“…Therefore, considering the shorter period of FSH support required for superstimulation before OPU [19,43] compared to traditional multiple ovulation and in vivo embryo procedures (reviewed by Mapletoft and Bó [36]), the use of a single injection of pFSH in 0.5% HA (5 mg/mL) might be an alternative to simplify the superstimulatory protocol for OPU-IVP programs. In a preliminary study performed in beef donor cows, the administration of 160 mg of pFSH diluted in 0.5% HA resulted in a comparable number of cumulus-oocyte complexes (COCs) recovered and blastocysts produced as the administration of 160-mg pFSH in twice-daily IM injections [44].…”

<b>Suplementação exógena com gonadotrofinas para aumentar a produção <i>in vitro</i> de embriões em doadoras Holandesas</b>

“…Although a two injection protocol seemed to be required to superovulate donor cows for in vivo embryo production, the results of the present study support the notion that only one injection of pFSH diluted in 0.5% HA is required for in vitro embryo production following OPU in non-lactating Holstein donors. These results are also in agreement with those obtained after a single injection of pFSH diluted in 0.5% HA in beef cows (ONGARATTO et al, 2011). The present data verify a shortened, but extended period of elevated plasma concentrations of FSH in HA groups.…”

“…Therefore, considering the shorter period of FSH support required for superstimulation prior to OPU (BLONDIN et al, 2002;VIEIRA et al, 2014a) compared to traditional multiple ovulation and in vivo embryo procedures [reviewed by Mapletoft and Bó (2012)], the use of a single injection of pFSH in 0.5% HA (5 mg/mL) might be an alternative to simplify the superstimulatory protocol for OPU-IVP programs. In a preliminary study performed in beef donor cows, the administration of 160 mg of pFSH diluted in 0.5% HA resulted in a comparable number of COCs recovered and blastocysts produced as the administration of 160 mg pFSH in twice daily IM injections (ONGARATTO et al, 2011).…”

Ovum Pick Up (Opu) by Transvaginal Ultrasonography in Cattle

“…Since the ultimate goal of pre-OPU superstimulation is to generate additional follicles rather than inducing multiple ovulations, changes in the dose and timing of gonadotropins used are required (Bols and Stout 2018). For this purpose, many methods have been tried in pre-OPU superstimulation protocols, such as using a single dose of FSH, different administration routes, reducing the amount of FSH and dissolving it in different polymers (Bó et al 1994;Chaubal et al 2007;Ongaratto et al 2010;Vieira et al 2016).…”