scholarly journals Effects of Extra-Long-Acting Recombinant Bovine FSH (bscrFSH) on Cattle Superovulation

Animals ◽  
2022 ◽  
Vol 12 (2) ◽  
pp. 153
Author(s):  
Miguel A. Gutiérrez-Reinoso ◽  
Constanza J. Aguilera ◽  
Felipe Navarrete ◽  
Joel Cabezas ◽  
Fidel O. Castro ◽  
...  

Over the last few years, several commercial FSH products have been developed for cattle superovulation (SOV) purposes in Multiple Ovulation and Embryo Transfer (MOET) programs. The SOV response is highly variable among individuals and remains one of the main limiting factors in obtaining a profitable number of transferable embryos. In this study, follicle stimulating hormone (FSH) from different origins was included in two SOV protocols, (a) FSH from purified pig pituitary extract (NIH-FSH-p; two doses/day, 12 h apart, four consecutive days); and (b) extra-long-acting bovine recombinant FSH (bscrFSH; a single dose/day, four consecutive days), to test the effects of bscrFSH on the ovarian response, hormone profile levels, in vivo embryo production and the pluripotency gene expression of the obtained embryos. A total of 68 healthy primiparous red Angus cows (Bos taurus) were randomly distributed into two experimental groups (n = 34 each). Blood sample collection for progesterone (P4) and cortisol (C) level determination was performed together with ultrasonographic assessment for ovarian size, follicles (FL) and corpora lutea (CL) quantification in each SOV protocol (Day 0, 4, 8, and 15). Moreover, FSH profiles were monitorised throughout both protocols (Day 0, 4, 5, 6, 7, 8, 9, 10, and 15). In vivo embryo quantity and quality (total structures, morulae, blastocysts, viable, degenerated and blocked embryos) were recorded in each SOV protocol. Finally, embryo quality in both protocols was assessed by the analysis of the expression level of crucial genes for early embryo development (OCT4, IFNt, CDX2, BCL2, and BAX). P4 and cortisol concentration peaks in both SOV protocols were obtained on Day 15 and Day 8, respectively, which were statistically different compared to the other time-points (p < 0.05). Ovarian dimensions increased from Day 0 to Day 15 irrespective of the SOV protocol considered (p < 0.05). Significant changes in CL number were observed over time till Day 15 irrespective of the SOV protocol applied (p < 0.05), being non- significantly different between SOV protocols within each time-point (p > 0.05). The number of CL was higher on Day 15 in the bscrFSH group compared to the NIH-FSH-p group (p < 0.05). The number of embryonic structures recovered was higher in the bscrFSH group (p = 0.025), probably as a result of a tendency towards a greater number of follicles developed compared to the NIH-FSH-p group. IFNt and BAX were overexpressed in embryos from the bscrFSH group (p < 0.05), with a fold change of 16 and 1.3, respectively. However, no statistical differences were detected regarding the OCT4, CDX2, BCL2, and BCL2/BAX expression ratio (p > 0.05). In conclusion, including bscrFSH in SOV protocols could be an important alternative by reducing the number of applications and offering an improved ovarian response together with better embryo quality and superior performance in embryo production compared to NIH-FSH-p SOV protocols.

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e12077
Author(s):  
Khalid Mahmood ◽  
Muhammad Zahid Tahir ◽  
Mahboob Ahmad Butt ◽  
Shazia Mansoor Qureshi ◽  
Amjad Riaz

Multiple Ovulation and Embryo Transfer (MOET) technology is a potential technique to upgrade livestock species’ genetics. The varied response to super-stimulatory treatments remains one of the limiting factors to this technology’s widespread use. The present study was aimed to improve the superovulation response and in-vivo embryo production by using controlled internal drug release (CIDR)-GnRH or CIDR-EB (Estradiol Benzoate) along with conventional superovulation protocol in Holstein Frisian (HF): Bos taurus; n = 42) and Crossbred (XB: Cholistani (Bos indicus) × HF; n = 28) cows. In the CIDR-GnRH/CIDR-EB treatment, CIDR was implanted in the cows after confirming the presence of a corpus luteum (CL) on the 8th day after estrus. 2 ml GnRH (Lecirelin acetate 0.0262 mg/ml) or 2 mg EB was also administered in CIDR-GnRH/CIDR-EB groups, respectively. Both groups were given super-stimulatory treatment from the 11th day after estrus (FSH in tapering doses twice a day for four consecutive days). On day 13, two doses of 2 ml prostaglandin (75 µg/ml of dextrorotatory cloprostenol) were administered (am: pm), and CIDR was removed the following day. Two artificial inseminations (AI) of the cows were performed (12 h apart) on the 15th day. No CIDR and GnRH/E.B were given in the control group, but the remaining superovulation protocol was the same. Later on, seven days after the first AI, non-surgical embryo flushing was done. The transferable embryos produced from three different superovulation protocols were then transferred into the recipient cows (n = 90) for determining their fertility. Statistical analysis revealed that the number of super-estrus follicles (SEF), multiple corpora lutea (MCL), ovulation/fertilization percentage, fertilized structures recovered (FSR), and transferable embryos (TEs) remained significantly higher (p < 0.05), and days taken for return to estrus (RTE) after embryo collection remained significantly lower (p < 0.05) in CIDR-GnRH (n = 18) and CIDR-EB (n = 15) groups as compared to the control (n = 37). The comparison between XB and HF cows revealed that the TEs production in CIDR-GnRH (XB = 5 vs HF = 13) and CIDR-EB (XB = 6 vs HF = 9) based superovulation protocols were 11.60  ±  4.08 vs 04.31  ±  0.98 and 09.33  ±  1.78 vs 05.22  ±  1.36, respectively. TEs production in XB cows (n = 5) of the CIDR-GnRH group was significantly higher (11.60  ±  4.08) than other groups. On the other hand, the days taken for RTE after embryo collection remained significantly lower (p < 0.05) in HF cows of treatment groups. However, the fertility of TEs was neither affected significantly (p > 0.05) by the superovulation protocol used nor by breed differences among donor cows. In conclusion, using CIDR-GnRH or CIDR-EB along with conventional superovulation protocol may enhance the efficiency of MOET programs in cattle. Furthermore, XB donor cows demonstrated a better performance than HF donor cows under subtropical conditions.


2021 ◽  
Author(s):  
STEFAN CIORNEI ◽  
Dan DRUGOCIU ◽  
Liliana Margareta Ciornei ◽  
Petru ROŞCA

Abstract BackgroundThe success of an embryo transfer protocol in sheep depends on many factors, but the choice of drugs for the desired superovulation as well as the conception rate are most essential. Reproductive activity in sheep is characterized by a seasonality influenced by several factors such as photoperiod, latitude, temperature, nutrition and breed. Reproductive seasonality and nutritional condition are the main factors that influence embryo production in sheep. In sheep, some anatomical peculiarities limit the application of traditional reproductive biotechnologies used in cattle. MethodsIn vivo embryo production is often referred to as “multiple ovulation and embryo transfer” and involves ovarian superstimulation of the donor female, insemination or mating, uterine flushing for embryo recovery, and either cryopreservation or transfer of collected embryos to recipients. A total number of 60 sheep and 3 rams were included in this study, divided into 2 groups (receptors/donors). Donor Suffolk sheep were treated for superovulation using the P4‐PGF‐FSH protocol while the cross-bred recipients’ group was synchronized with P4-PGF-PMSG. ResultsOn the first day after superovulation, all ovaries had more than 5 dominant follicles, while corpora lutea were later observed in 83.3% sheep. The recovery rate was 83.3% while 72,9% embryos were transferable. Embryos were transferred directly into recipients. Fertility after 30 days was 68.57%, lambing rate was 91.6%, and CR 62.85%. This study showed that veterinary drugs (P4, FSH, LH, PMSG, PGF) used for superovulation were capable to induce estrus and synchronize ovulation in sheep, are topical and in increasing use worldwide. ConclusionsThe aim of this study was to conclude on the effectiveness of a wider on farm in vivo embryo transfer development program in Suffolk sheep, using several veterinary hormones. The application of a multiple ovulation embryo transfer (MOET) protocol has a positive effect in the production of in vivo derived embryos in Suffolk sheep and can guarantee the success of embryo transfer activity to ewes with lower genetic merit. Our research aimed at representing a model for sheep farms for a rapid improvement of productive traits.


2021 ◽  
Vol 33 (2) ◽  
pp. 181
Author(s):  
C. Brochado ◽  
B. J. Duran ◽  
J. C. L. Motta ◽  
J. D. Kieffer ◽  
A. Pinczak ◽  
...  

The objective of the present study was to evaluate the effect of length of the FSH superstimulatory treatment on ovarian response and embryo production in sheep. Poll Dorset ewes (n=63) 3.2±0.2 years old weighing 58.5±1.5kg and with a body condition score of 2.7±0.1 (0=emaciated; 5=obese) were used during the transition from the breeding to the non-breeding season. All ewes received an ovarian superstimulatory treatment during the first follicular wave (Day 0 protocol). On Day −9 all ewes received a 0.3-g progesterone intravaginal device (CIDR, Zoetis) for 6 days. On Day −3, the CIDR was removed and all ewes were administered 125μg of cloprostenol sodium intramuscularly (IM) (PGF, estroPLAN, Parnell) and 200IU of equine chorionic gonadotrophin (eCG)+100IU of human chorionic gonadotrophin (hCG) IM (P.G. 600, Merck). Thirty-six hours after CIDR removal (Day −1.5) ewes were administered 100μg of gonadorelin acetate IM (gonadotrophin-releasing hormone, GnRH; Gonabreed, Parnell). Superstimulatory treatments were initiated on Day 0 (84 h after CIDR removal) with a total of 240mg of NIH-FSH-P1 (12 mL, Folltropin, Vetoquinol), and a new CIDR was used from the first to the last FSH dose (6-dose=3 days; 8-dose=4 days). Ewes were randomised to receive the total dose of FSH distributed in eight decreasing doses (8-dose group, n=33) or six decreasing doses (6-dose group, n=30) twice daily 12h apart. Two doses of PGF were administered with the last two doses of FSH, and GnRH was administered 12h after the last FSH administration. Intrauterine AI with fresh semen was performed by laparoscopy 16h after GnRH administration. Ovarian response was determined by laparoscopy 6 days after AI and ova/embryos were collected surgically in ewes with &gt;2 corpora lutea (CL). Data were analysed using generalized linear mixed models (SAS 9.4; SAS Institute Inc.) and presented as mean±s.e.m. in Table 1. The percentage of responding donors (&gt;2 CL) was 93.3% (28/30) and 78.8% (26/33; P=0.5) for the 6-dose and 8-dose groups, respectively. The number of CL tended (P=0.06) to be lesser in ewes in the 6-dose than the 8-dose group. However, no differences (P&gt;0.10) were found in total ova/embryo, fertilized ova, transferable embryos, or freezable embryos. In conclusion, despite a tendency for a greater ovarian response in ewes treated with 8 doses of FSH, embryo yield did not appear to differ after either 6 or 8 doses of FSH administered during the first follicular wave in sheep. Table 1. Ovulatory response and embryo production in responding ewes (&gt;2 corpora lutea) superstimulated with either 6 or 8 doses of FSH during the first follicular wave Outcome Treatment P-value 6-dose 8-dose Number of ewes 28 26 Corpora lutea (n) 14.2±1.7 20.3±2.8 0.06 Total ova/embryos (n) 9.0±1.3 10.9±1.8 0.38 Fertilized ova (n) 8.1±1.4 10.2±1.9 0.34 Fertilized ova (% per ewe) 89.0±5.0 81.7±7.0 0.40 Degenerate embryos 1.3±0.4 2.0±0.8 0.89 Transferable embryos (Grade 1–3) 6.8±1.2 8.2±1.8 0.52 Transferable embryos (% per ewe) 85.7±4.4 75.4±8.4 0.46 Freezable embryos (Grade 1–2) 6.6±1.1 8.2±1.8 0.43 Freezable embryos (% per ewe) 80.5±5.6 75.4±8.4 0.61


2015 ◽  
Vol 36 (6Supl2) ◽  
pp. 4215
Author(s):  
João Bosco Loiola Filho ◽  
Alane Pains Oliveira do Monte ◽  
Thais Thatiane Dos Santos Souza ◽  
Mayara De Souza Miranda ◽  
Lívia Correia Magalhães ◽  
...  

To evaluate the effect of pFSH dose on the in vivo embryo production of Dorper ewes in the semi-arid northeast of Brazil, 40 sheep females were distributed into two groups of 20 animals that received intravaginal CIDR for 14 days, and two days before device removal, they received one of the following treatments: in the FSH200 group, the ewes received 200 mg of pFSH; and in the FSH128 group, the ewes received a total of 128 mg in decreasing doses every 12 h. Beginning 12 h after the conclusion of the treatments, estrus detection was performed every four hours using two Dorper rams of proven fertility. The ewes were mated at estrus onset and 24 hours later. Seven days after intravaginal device removal, the superovulatory response was evaluated, and embryo collection was performed using the laparotomy method. The recovered flushings were subjected to embryo searches under a stereomicroscope and classified according to their qualities. Analyses of variance (ANOVAs) and LSD tests were used to compare the different parameters. The data expressed as percentages were analysed by chi-square test. The ovulation rate was higher in the FSH200 group, which had 16.3 ± 0.3 corpora lutea (CL), than in the FSH128 group, which had 11.3 ± 0.3 CL (P<0.05). However, higher fertilization rate (83.6% vs. 62.4%) and higher transferable (86.0% vs. 71.6%) and freezable (67.9% vs. 40.8%) embryo rates were observed in the FSH 128 group compared with the group that received 200 mg. Furthermore, no significant differences in the remaining parameters were observed between the experimental groups (P>0.05), demonstrating that pFSH dose reduction promoted a greater production of freezable and transferable embryos in Dorper ewes subjected to MOET.


2015 ◽  
Vol 36 (6Supl2) ◽  
pp. 4215 ◽  
Author(s):  
João Bosco Loiola Filho ◽  
Alane Pains Oliveira do Monte ◽  
Thais Thatiane Dos Santos Souza ◽  
Mayara De Souza Miranda ◽  
Lívia Correia Magalhães ◽  
...  

<p>To evaluate the effect of pFSH dose on the <em>in vivo </em>embryo production of Dorper ewes in the semi-arid northeast of Brazil, 40 sheep females were distributed into two groups of 20 animals that received intravaginal CIDR for 14 days, and two days before device removal, they received one of the following treatments: in the FSH200 group, the ewes received 200 mg of pFSH; and in the FSH128 group, the ewes received a total of 128 mg in decreasing doses every 12 h. Beginning 12 h after the conclusion of the treatments, estrus detection was performed every four hours using two Dorper rams of proven fertility. The ewes were mated at estrus onset and 24 hours later. Seven days after intravaginal device removal, the superovulatory response was evaluated, and embryo collection was performed using the laparotomy method. The recovered flushings were subjected to embryo searches under a stereomicroscope and classified according to their qualities. Analyses of variance (ANOVAs) and LSD tests were used to compare the different parameters. The data expressed as percentages were analysed by chi-square test. The ovulation rate was higher in the FSH200 group, which had 16.3 ± 0.3 corpora lutea (CL), than in the FSH128 group, which had 11.3 ± 0.3 CL (P&lt;0.05). However, higher fertilization rate (83.6% vs. 62.4%) and higher transferable (86.0% vs. 71.6%) and freezable (67.9% vs. 40.8%) embryo rates were observed in the FSH 128 group compared with the group that received 200 mg. Furthermore, no significant differences in the remaining parameters were observed between the experimental groups (P&gt;0.05), demonstrating that pFSH dose reduction promoted a greater production of freezable and transferable embryos in Dorper ewes subjected to MOET.</p>


2014 ◽  
Vol 26 (4) ◽  
pp. 527 ◽  
Author(s):  
Ricardo S. Surjus ◽  
Alexandre B. Prata ◽  
Marta Borsato ◽  
Fernanda C. S. Z. Mattos ◽  
Mariana C. Martins da Silveira ◽  
...  

The present study evaluated superovulatory responses and in vivo embryo production in cows treated with FSH starting 1 or 2 days after ovum pick-up (OPU). Thirty-three non-lactating Nelore cows were subjected to aspiration of all follicles ≥3 mm for OPU. After OPU, cows were randomly divided into two groups in which the follicle superstimulatory treatments with FSH started 1 or 2 days after OPU (Groups D1 and D2, respectively). Data are presented as the least squares mean ± s.e.m. The number of follicles ≥3 mm before OPU was similar between groups (~34); however, cows in Group D2 had more follicles ≥3 mm on the first day of FSH (15.2 ± 2.3 vs 7.6 ± 1.7; P = 0.04) and a higher ratio of the number of follicles at first FSH/number of follicles before OPU (0.41 ± 0.04 vs 0.24 ± 0.02; P = 0.01). In addition, Group D2 cows had a greater superovulatory response than did cows in Group D1 (18.9 ± 2.8 vs 9.1 ± 1.9 corpora lutea, respectively; P < 0.03). However, there was no difference in the total number of recovered ova and embryos from cows in Groups D2 and D1 (5.1 ± 1.4 vs 4.9 ± 1.3, respectively; P > 0.10). Nevertheless Group D2 cows had more freezable embryos than Group D1 cows (3.2 ± 1.1 vs 1.3 ± 0.5, respectively; P < 0.05). Cows from Group D2 had a much higher proportion (P < 0.001) of follicles ≥8 mm compared with follicles ≥6 mm and <8 mm at the time of the last treatment with FSH. In conclusion, to obtain a greater production of viable embryos in superovulated cows after OPU, it is recommended to wait at least 2 days before starting FSH treatment.


2013 ◽  
Vol 25 (1) ◽  
pp. 268
Author(s):  
M. Kaymaz ◽  
A. R. Agaoglu ◽  
K. Karakas ◽  
I. Pir Yagci ◽  
O. Korkmaz Agaoglu ◽  
...  

The Angora, Kilis, Honamli, Hair, and Norduz are native goat breeds in Turkey and are currently in danger of extinction. This study aimed to assess the efficacy of the repeated administration of a superovulatory (SOV) protocol for in vivo embryo production in these breeds. A total of 14 Angora, 15 Kilis, 10 Honamli, 10 Hair, and 9 Norduz goats were used in this work. The synchronization procedure was started on Day 5 after visible oestrus by using controlled internal drug release dispensers (CIDR®) for 11 days. Administration of FSH (Folltropin®) began on Day 9 (twice daily) and continued for 3 days (total dose: 200 mg; 50 mg × 2.30 mg × 2.20 mg × 2). A dose of prostaglandin F2α (1.6 mg; Dalmazin®) was injected together with first FSH injection. Gonadotropin-releasing hormone (Receptal®: 100 µg) was injected 6 h before mating. All goats in oestrus were naturally mated twice a day for 3 days. Ovarian examination (number of corpora lutea) and embryo recovery were performed by laparotomy on Day 6 after CIDR® withdrawal. Each uterine horn was flushed, and the embryos were recovered and counted. To avoid intra-abdominal adhesions, a 2.5% heparin solution was used during flushing. The SOV procedure was repeated once per year during the breeding season (2009 to 2011). Fertilization and recovery rates were calculated. Differences in the SOV response and embryo yields were evaluated by Friedman’s test. In Hair goats, the number of corpora lutea decreased significantly (P < 0.05) during the third SOV cycle (12.7 ± 6.2, 14.0 ± 9.1, and 6.8 ± 5.6, respectively, for cycles 1, 2, and 3), whereas no effect of the cycle was observed in the remainder of breeds. The number of expanded blastocysts increased considerably during the third cycle in Angora (0, 0.2 ± 0.8, and 1.4 ± 2.9, respectively, for cycles 1, 2, and 3), Kilis (0.2 ± 0.4, 0.3 ± 1.3, and 4.2 ± 5.0), and Honamli (0, 1.3 ± 1.7, and 3.6 ± 4.5) goats, whereas a significant decrease was observed in Norduz goats (2.4 ± 5.0, 1.8 ± 2.0, and 0.1 ± 0.3; P < 0.05). The mean numbers of unfertilized oocytes were found to be significantly increased in Angora (0.4 ± 1.6, 0, and 2.1 ± 4.1, respectively, for cycles 1, 2, and 3), Kilis (0, 1.3 ± 3.9, and 3.1 ± 5.2), and Honamli (0, 4.9 ± 5.2, and 4.5 ± 7.8) goats (P < 0.05). As a result, fertilization rates (%) showed a decrease in Angora (50, 100, 24.5, respectively, for cycles 1, 2, and 3) and Honamli (100, 42.5, and 56.3) goats (P < 0.05), whereas recovery rates showed no difference among the different breeds. The methodology presented in this study was found to be an efficient technique for superovulation of the Angora, Kilis, and Honamli goats. Because Hair and Norduz are relatively small breeds, the dosage of FSH might have had a negative effect on the superovulation and embryo yield. Additionally, the use of intra-abdominal washing solutions for preventing adhesions as observed in previous works (data not shown) is believed to have a positive effect on achieving high levels of efficiency in in vivo embryo production.


2013 ◽  
Vol 25 (1) ◽  
pp. 302
Author(s):  
J. M. Palomino ◽  
R. J. Mapletoft ◽  
M. Anzar ◽  
M. R. Woodbury ◽  
M. P. Cervantes ◽  
...  

Superovulation protocols are being developed in wood bison (Bison bison athabascae), a threatened Canadian species. In initial studies, 2 doses of FSH diluted in hyaluronan given 48 h apart were successful for inducing ovarian superstimulation in wood bison, and ovulation rate was improved by final treatment with hCG instead of LH (Palomino et al. 2012 Reprod. Fertil. Dev. 24, 226). In that study, exogenous progesterone had no effect on the number of ovulations, but embryo quality could not be evaluated because of the low number of embryos collected. In beef cattle, replacement of the final doses of FSH with eCG has resulted in the recovery of a greater number of ova/embryos. The objective of the present study was to evaluate the effect of exogenous progesterone (PRID) on embryo quality and to determine if the addition of eCG increases the ovulation rate in superstimulated wood bison. Follicular ablation was done to synchronize follicular wave emergence in 26 wood bison cows during the anovulatory season (May). Cows were assigned randomly to 3 groups: PRID/no eCG (n = 8), PRID/eCG (n = 9), and no PRID/eCG (n = 9). A PRID was inserted on the day of follicular ablation (Day –1) in respective groups. In all bison, FSH diluted in hyaluronan (5 mg mL–1, MAP-5, Bioniche Animal Health Inc., Belleville, Ontario, Canada) was given intramuscularly on Day 0 (300 mg) and Day 2 (100 mg). On Day 3, a single dose of 450 IU of eCG (Pregnecol, Bioniche Animal Health Inc.) was given intramuscularly and the PRID were removed in the corresponding groups. On Day 5, all bison were given 2500 IU of hCG (Chorulon, Merck, Whitehouse Station, NJ, USA) intramuscularly to induce ovulation. Bison were inseminated with chilled semen 12 and 24 h later. Nonsurgical embryo collection was performed on Day 14. Transrectal ultrasonography was done to record the ovarian response, ovulation rate, and number of corpora lutea (CL). Results were compared by ANOVA and Chi-squared test (Table 1). The number of ovulatory-sized follicles (≥9 mm) on Day 5 did not differ among groups. Ovulation rate was lowest in bison treated with both a PRID and eCG (P < 0.05). There were no differences among groups in the number of CL on Day 14, the number of ova/embryos collected, or the number of transferable embryos. The superovulatory response and embryo collection rate in the present study were higher than in any previous reports in bison. The ovulation rate was not improved by the addition of eCG treatment, and exogenous progesterone had no effect on embryo quality. Table 1.Response of superstimulated wood bison (mean ± SEM) to treatment with exogenous progesterone (PRID) and/or eCG


2017 ◽  
Vol 29 (1) ◽  
pp. 208 ◽  
Author(s):  
R. Fry

The administration of sustained-release FSH-MAP5 in a 2-injection protocol has been shown to be as effective as a multiple-FSH injection protocol in inducing superovulation in cattle (Bo and Mapletoft 2014 Theriogenology 81, 38) and in sheep (Fry et al. 2016 Reprod. Fertil. Dev. 28, 250); however, the effect on embryo quality in the latter experiment was unclear. The following experiment further investigated the effect of FSH-MAP5 on ovulation rate and embryo quality in a sheep multiple-ovulation embryo transfer (MOET) program conducted in the breeding season. Two hundred sixteen Dohne merino ewes received a 12-day CIDR-S device containing 0.33 g of progesterone (Zoetis, Florham Park, NJ, USA) plus 200 mg of FSH IM (Folltropin-V; Vetoquinol, Belleville, Canada) and 400 IU of eCG IM (Pregnecol: Vetoquinol) in 4 treatment groups. Group 1 (n = 51) received 7 injections (a.m., p.m.) of FSH in saline (2.5, 2.0, 1.5, 1.5, 1.0, 1.0, and 0.5 mL) starting 2.5 days before CIDR withdrawal and 400 IU of eCG in saline at the time of the first FSH injection. Group 2 (n = 53) received 6 injections (a.m., p.m.) of FSH in saline (3.0, 2.0, 1.5, 1.5, 1.0, and 1.0 mL) starting 2.5 days before CIDR withdrawal and 400 IU of eCG in saline at CIDR withdrawal. Group 3 (n = 56) received 3.3 mL of FSH in hyaluronan (50 mg of MAP5, Vetoquinol) and 400 IU of eCG in saline 2.5 days before CIDR withdrawal and 1.7 mL of FSH-MAP5 at 0.5 days before CIDR withdrawal. Group 4 (n = 56) received 3.3 mL of FSH-MAP5 at 2.5 days before CIDR withdrawal, 1.7 mL of FSH-MAP5 at 0.5 days before CIDR withdrawal, and 400 IU of eCG in saline at CIDR withdrawal. Ewes were inseminated with semen collected and pooled from 5 rams at 36 to 40 h after CIDR withdrawal. Donor ewes were slaughtered 6 days after AI, corpora lutea were counted on the ovary, and ova/embryos were collected. Data for corpora lutea, total ova/embryo, and transferable embryo was analysed by the Kruskal–Wallis test and differences between groups were determined by the Dunn’s test. Results are shown in Table 1. There were no differences in the mean number of ovulations or total ova/embryo collected between any group (P > 0.05); however, those receiving eCG at the time of CIDR withdrawal (Groups 2 and 4) had significantly more transferable embryos than those receiving eCG at the time of the first FSH injection (Groups 1 and 3). In conclusion, the administration of the sustained-release FSH-MAP5 in a 2-injection protocol in sheep was as effective as a multiple-FSH injection protocol in inducing multiple ovulations and yielded similar number of transferable embryos, whereas the administration of eCG at the time of CIDR withdrawal resulted in more transferable embryos, possibly by improving fertilisation rates. Table 1. Results1


Reproduction ◽  
2003 ◽  
pp. 101-111 ◽  
Author(s):  
BK Campbell ◽  
DT Baird ◽  
CJ Souza ◽  
R Webb

The aim of this study was to differentiate between pituitary and ovarian actions of the FecB gene by measuring the ovarian response to a standardized treatment with gonadotrophins designed to mimic the changes in FSH and LH that occur in the follicular phase of the ovarian cycle in ewes, with (Fec(B/-), n=6) and without (Fec(+/+), n=9) the gene, that were rendered hypogonadotrophic by pretreatment with a potent antagonist of GnRH. Ewes with ovarian autotransplants were used to facilitate the assessment of follicular function by the collection of ovarian venous blood and ultrasonography. The gonadotrophin regimen resulted in concentrations of FSH and LH that were similar to concentrations found in a normal cycle and did not differ between genotypes. Follicular development and ovulation occurred in all animals, and patterns of secretion of oestradiol, androstenedione and inhibin A were normal. Despite these endocrine similarities, the antral follicle population stimulated by FSH infusion retained the characteristic genotypic difference with the ovaries of Fec(+/+) animals containing a range of follicle sizes with decreasing proportions of small (<3.5 mm in diameter) and medium (3.5-4.5 mm in diameter) follicles as well as large follicles (> or =4.5 mm in diameter), whereas the ovaries of Fec(B/-) ewes contained no follicles of >4.5 mm in diameter. This genotypic difference was retained after ovulation with gene carriers having more preovulatory follicles/corpora lutea (3.8+/-0.3) of a smaller diameter (5.3+/-0.3 mm) than did non-gene carriers (1.7+/-0.3; 11.4+/-0.9 mm; P<0.05). As ewes carrying the FecB gene mutation were able to ovulate more follicles than non-gene carriers, despite identical concentrations and patterns of FSH and LH stimulation, the results of this study support the hypothesis that the FecB gene acts at the ovary to enhance ovarian sensitivity to gonadotrophic stimulation.


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