New approaches to superovulation and embryo transfer in small ruminants

2010 ◽  
Vol 22 (1) ◽  
pp. 113 ◽  
Author(s):  
A. Menchaca ◽  
M. Vilariño ◽  
M. Crispo ◽  
T. de Castro ◽  
E. Rubianes

The present paper reviews the current state of knowledge in multiple ovulation and embryo transfer (MOET) technology in small ruminants, focusing on recently reported information. Major new findings are related to follicular wave patterns in small ruminants, the elucidation of follicular dominance and the integration of this information into ovarian superstimulatory treatment protocols. Follicular dynamics determine steroid and gonadotrophin secretion, follicular responses to FSH, ovulatory responses and embryo yield. Protocols that control follicular dominance have been designed to allow the initiation of superstimulation at the beginning of a follicular wave. New approaches consist of (1) synchronisation of ovulation for superstimulation during Wave 1 (i.e. Day 0 protocol), (2) pretreatment with a gonadotrophin-releasing hormone (GnRH) antagonist from 10 days prior to FSH treatment to avoid follicular dominance and (3) progesterone–oestradiol cotreatment to synchronise follicle wave emergence. These protocols provide a homogeneous pool of small follicles that are gonadotrophin responsive, enhancing the superovulatory response and embryo yield with a reduction in the incidence of unovulated follicles and early regression of corpora lutea. In addition, the rate of fertilisation failure has been reduced by using an inducer of ovulation (i.e. GnRH) associated with intrauterine insemination. In summary, the application of recently acquired knowledge has resulted in relevant improvements in MOET programmes in small ruminants.

2001 ◽  
Vol 49 (4) ◽  
pp. 431-441 ◽  
Author(s):  
S. Cseh ◽  
L. Solti

The objectives of this study were (a) to assess the ovulatory response and embryo production of Hungarian Merino ewes after superovulation, (b) to investigate the factors influencing the efficiency of embryo transfer (ET) in Hungarian Merino ewes, (c) to compare the results of two ovarian stimulation protocols (PMSG and PMSG+FSH treatment) in Hungarian Merino ewes, and (d) to study how superovulation, laparoscopic insemination and surgical embryo retrieval (ER) affect the subsequent reproduction of Hungarian Merino donor females after an ET programme. There was no significant difference between the ovarian stimulation protocols in the ratio of donor ewes responding to superovulation nor in the average number of corpora lutea. However, the number of transferable embryos recovered per donor ewe was higher in the PMSG+FSH group. The proportion of transferable embryos, unfertilised oocytes and degenerated embryos did not differ between the treatment protocols. The total pregnancy rate was 53.4% (179/335). Neither the developmental stage of the embryo nor the number of transferred embryos affect the implantation of embryos. However, the increased number of transferred embryos positively influenced the pregnancy rate. No difference was found in the pregnancy rate between synchronised and nonsynchronised groups of recipients. Thirty-six out of 45 donor ewes (80%) became pregnant within one year after the ET programme, indicating that ovarian stimulation and surgical ER did not affect adversely their reproduction.


2012 ◽  
Vol 24 (1) ◽  
pp. 228
Author(s):  
M. E. F. Oliveira ◽  
C. C. D'Amato ◽  
L. G. Oliveira ◽  
S. D. Bicudo ◽  
J. F. Fonseca ◽  
...  

This study was designed to investigate if the superovulatory response and embryo yield in Santa Ines ewes are influenced by the time that FSH treatments are inititiated: near the emergence of the first or last follicular wave of a progesterone-based (P4) protocol. Days of emergence of the follicular waves was defined in a previous study that evaluated follicular dynamics in oestrus synchronization treatments (Oliveira et al. 2011 Acta Sci. Vet. 40). Twenty Santa Ines cyclic ewes were submitted to 2 superovulatory protocols according to the time that FSH treatments were initiated (G-first wave, n = 10 and G-last wave, n = 10). Thus, the FSH treatment started on Day 4 and 10 of protocol for G-first and G-last, respectively (Day 0 = onset of protocol). Ewes were treated with a P4 device (CIDR®; Pfizer-New Zealand) for 6 and 12 days for G-first and G-last, respectively. Additionally, in G-last, the CIDR was replaced by a new one on Day 7. All ewes received 2 injections of 37.5 μg of D-cloprostenol (Prolise®, Arsa-Argentina) on Day 0 and at CIDR removal. The superovulatory regimen consisted of 8 IM injections of pFSH (Folltropin®; Bioniche Animal Health, Belleville, Ontario, Canada) administrated twice daily (40, 40, 30, 30, 20, 20, 10 and 10 mg of porcine FSH). A single IM dose of 200 IU of eCG (Novormon®, Syntex, Buenos Aires, Argentina) was given concurrently with CIDR removal. Ewes were mated by a fertile ram. Embryo collections were accomplished 7 days after CIDR removal. The superovulatory response was classified in scores: (0) 4 or fewer corpora lutea (CL); (1) between 5 and 10 CL and (2) 11 or more CL. Means (± SD) were compared using Kruskal-Wallis test and percentages using Chi-square (P < 0.05). Most donors (70%, 7/10) from G-last had a superovulatory response classified as score 2 and the remainder (30%, 3/10) as score 1, whereas half of ewes from G-first were classified as score 2 and half as score 1 (P > 0.05).There was no effect between treatments (P > 0.05) for ovulation rate (G-first: 90.7 ± 10.2% vs G-last: 93.0 ± 7.2%), number of ovulations (G-first: 13.1 ± 7.3 vs G-last: 14.0 ± 5.8), or number of luteinized unovulated follicles (G-first: 1.4 ± 1.3 vs G-last: 0.9 ± 1.0). Furthermore, there was no effect between G-first and G-last (P > 0.05) in the rate of ova/embryos recovered (55.1 ± 27.1% vs 67.0 ± 19.4%), mean number of ova/embryos recovered (7.0 ± 5.5 vs 9.1 ± 3.8), number of viable embryos (3.9 ± 6.1 vs 5.7 ± 4.4), or viability rate (49.1 ± 43.7 vs 58.5 ± 34.6). In conclusion, superovulatory response and embryo yield did not differ between FSH treatments initiated during the first or last follicular wave of a progesterone-based treatment in Santa Ines ewes. Financial support of FAPESP is acknowledged.


2012 ◽  
Vol 24 (1) ◽  
pp. 278 ◽  
Author(s):  
Reuben J. Mapletoft ◽  
Gabriel A. Bó

Superovulation protocols have improved greatly since the early days of bovine embryo transfer when purified gonadotrophins were not available, follicular wave dynamics were unknown physiological phenomena and prostaglandins were not available. Although superstimulatory protocols in cattle are normally initiated mid-cycle, elective control of follicular wave emergence and ovulation have had a great impact on the application of on-farm embryo transfer. However, the most common treatment for the synchronisation of follicular wave emergence involves the use of oestradiol, which cannot be used in many parts of the world. Therefore, the need for alternative treatments has driven recent research. An approach that has shown promise is to initiate follicle-stimulating hormone (FSH) treatments at the time of the emergence of the new follicular wave following ovulation induced by gonadotrophin-releasing hormone. Alternatively, it has been shown that it may be possible to ignore follicular wave status and, by extending the treatment protocol, induce subordinate follicles to superovulate. Finally, the short half-life of pituitary FSH necessitates twice-daily treatments, which are time-consuming, stressful and subject to error. Recent treatment protocols have permitted superstimulation with a single FSH treatment or two treatments 48 h apart, reducing the need for animal handling during gonadotrophin treatments.


2013 ◽  
Vol 25 (1) ◽  
pp. 306
Author(s):  
M. E. F. Oliveira ◽  
C. C. D'Amato ◽  
C. S. Oliveira ◽  
F. F. P. C. Barros ◽  
A. P. Perini ◽  
...  

This study was designed to investigate if the time of onset of FSH treatment [near the emergence of first or last follicular wave on progesterone (P4) protocol] influenced the superovulatory response and embryo yield in Santa Ines ewes during breeding season. Days of emergence of the follicular waves were defined in a previous study that evaluated the follicular dynamic in oestrus synchronization treatments (Oliveira et al. 2011 Acta Sci. Vet. 40). We observed emergence of the first and last follicular wave on 5.69 ± 0.42 and 11.25 ± 0.39 days of protocol, respectively. Twenty Santa Ines ewes were submitted to 2 superovulatory protocols according to the time that FSH treatments were initiated (G-first wave, n = 10; G-last wave, n = 10). On Day 0, all ewes received a P4 device (CIDR®; Pfizer Animal Health, New York, NY, USA) and injection of 37.5 µg of d-cloprostenol, IM. The FSH treatments started on Day 6 and Day 11 of protocol for G-first and G-last, respectively. The superovulatory regimen consisted of 8 IM injections of pFSH administrated twice daily (40, 40, 30, 30, 20, 20, 10, and 10 mg of pFSH). The P4 device was removed on Day 8 and Day 13 for G-first and G-last, respectively. At these times, all ewes received another injection of 37.5 µg of d-cloprostenol and a dose of 200 IU of eCG. During 4 days after the P4 device removal, ewes were mated by a fertile ram. Embryo collections were accomplished 7 days after CIDR withdrawal. The ovaries were evaluated by ultrasonography (3 times daily, during the mating period) and laparotomy (concomitantly with embryo collection). The superovulatory response was observed by classified by score: 0 = 4 or fewer corpora lutea (CL); 1 = between 5 and 10 CL; and 2 = 11 or more CL. Data were analysed by GLIMMIX using SAS software (SAS Institute Inc., Cary, NC, USA). All donors from G-first had superovulatory response classified as score 2, whereas 60% of ewes from G-last were classified as score 2, 20% as score 1, and 20% as score 0 (P < 0.05). There were effects between treatments (P < 0.05) for ovulation rate (G-first: 97.9 ± 1.4% v. G-last: 88.5 ± 4.4%) and number of ovulations (G-first: 17.0 ± 2.3 v. G-last: 12.5 ± 2.6). The numbers of luteinized unovulated follicles were 0.7 ± 0.5 for G-first and 1.2 ± 0.4 for G-last (P > 0.05). There was no difference between G-first and G-last (P > 0.05) in the rate of ova/embryos recovered (54.9 ± 5.7% v. 49.3 ± 8.5%), mean number of ova/embryos recovered (9.0 ± 1.4 v. 6.3 ± 1.1), number of viable embryos (3.8 ± 1.5 v. 3.4 ± 0.8), or viability rate (40.3 ± 10.8 v. 53.4 ± 12.1). In conclusion, the FSH treatment started near the emergence of the first follicular wave of progesterone protocol in Santa Ines ewes during the breeding season resulted in a higher superovulatory response than that started near the last follicular wave; however, no improvements in embryo yield were observed. Financial support: FAPESP.


2010 ◽  
Vol 22 (1) ◽  
pp. 132 ◽  
Author(s):  
Ciro M. Barros ◽  
Ronaldo L. Ereno ◽  
Renato A. L. Simões ◽  
Paulo Fernandes ◽  
José Buratini ◽  
...  

Embryo transfer is a biotechnology that has been used worldwide to increase the production of offspring from female bovines. Treatments to induce multiple ovulations (superovulation) have evolved from superstimulatory protocols that depended upon detection of oestrus to treatments that synchronise follicle growth and ovulation, allowing for improved donor management and fixed-timed AI (FTAI). The protocols associated with FTAI facilitate animal handling and produce at least as many viably embryos as conventional treatment protocols that required detection of oestrus. Recent knowledge regarding LH receptors (LHR) and follicular development can be applied to improve embryo transfer protocols. In fact, improvements in the superstimulatory treatment called the ‘P-36 protocol’, which include hormones that stimulate LHR, indicate that adjustments related to LHR availability may increase bovine embryo yield compared with conventional protocols based on the detection of oestrus.


2019 ◽  
Vol 31 (1) ◽  
pp. 17 ◽  
Author(s):  
Jeferson F. Fonseca ◽  
Maria Emilia F. Oliveira ◽  
Felipe Z. Brandão ◽  
Ribrio I. T. P. Batista ◽  
Alexandre R. Garcia ◽  
...  

Brazil has presented tremendous progress in non-surgical embryo transfer (NSET) in sheep and goats. New instruments and techniques for non-surgical embryo recovery (NSER) and NSET in small ruminants were implemented. Recent improvements include refinement of the protocols for cervical relaxation combining oestradiol–oxytocin–cloprostenol treatment at specific times before NSER in sheep; recipient goats do not require any hormonal drugs to induce cervical dilation and direct embryo transfer by the cervical route yields excellent results. Transrectal ovarian ultrasonography (B-mode but especially colour Doppler) have proven to be accurate methods to localise and enumerate corpora lutea and luteinised unovulated follicles in recipient and donor does and ewes. An array of new criteria for selecting superior animals for NSER and NSET (e.g. cervical mapping) have been developed by Brazilian researchers. Extensive studies on both technologies were initially conducted in commercial breeds of goats and sheep but have been gradually extended to some native breeds of sheep (germplasm conservation) and dairy goat operations. It is speculated that, in future, NSER and NSET may become methods of choice for caprine and ovine embryo recovery and transfer in Brazil, and then globally. Due primarily to the efficiency of NSET in goats, a novel interspecies (e.g. bovine) IVP method may soon be developed on a large scale. The Brazilian experience is an invaluable source of information and know-how promoting the replacement of conventional surgical assisted reproductive technologies with non-surgical procedures and hence supporting the rapid development of the embryo transfer industry in small ruminants.


2012 ◽  
Vol 24 (1) ◽  
pp. 226
Author(s):  
I. Singh ◽  
R. K. Sharma ◽  
S. K. Phulia ◽  
D. Kumar ◽  
O. Saxena ◽  
...  

Effect of the removal of inhibitory consequences of a dominant follicle (DF) on the recruitment, growth and ovulation of follicles in cyclic buffaloes induced to superovulate, was evaluated. Eight elite, multiparous, donor Murrah buffaloes, 80 to 120 days postpartum, were included in an integrated single-ovulation embryo transfer–multiple-ovulation embryo transfer (SOET-MOET) schedule. Superovulation with FSH (Folltropin® NIH-FSH-P1, Bioniche Animal Health, Belleville, ON, Canada) was preceded by nonsurgical single embryo (SOET) recovery on Day 5 or 6 after prostaglandin F2α (Lutalyse®, Pfizer, Madison, NJ, USA)-induced oestrus and AI (Day 0). For MOET, Folltropin was administered from Day 9 p.m. through to Day 13 p.m. in a twice-daily descending dose schedule (0–5, 5–4, 4–3, 3–2, 2–1 mL; total dose of 580 mg of NIH-FSH-P1). A dose of 25 mg of Lutalyse was administered on Day 12 a.m. and p.m. Donors were inseminated with proven semen on Day 14 a.m. and p.m. and Day 15 a.m. Nonsurgical embryo collection was carried out on Day 19 or 20 followed by administration of Lutalyse on Day 25. Following sexual rest for one complete oestrous cycle, Day 5 or 6 SOET was repeated. On Day 9, ultrasound-guided follicle aspiration was used to ablate the DF and the second MOET treatment was initiated at the same time. Mean diameter of DF at the start of superovulation treatment on Day 9 was similar between the first (12.5 ± 0.66 mm) and second (12.2 ± 0.33 mm) MOET cycles. At AI, mean number (9.6 ± 1.43 vs 13.4 ± 1.87) of follicles >9 mm were higher (P < 0.05) in the second (ablation) MOET cycle, though their mean diameters were similar (11.7 ± 0.21 vs 12.3 ± 0.21 mm; P = 0.17). On the day of ova/embryo collection, almost similar numbers of large follicles (4.1 ± 0.93 and 4.4 ± 0.84), with mean diameters of 12.9 ± 0.74 and 13.4 ± 0.83 mm, were observed for the first and second MOET, respectively. Mean (± SEM) numbers of corpora lutea (3.9 ± 0.81 and 4.9 ± 0.61; P = 0.19) and transferable quality embryos (0.6 ± 0.37 and 1.6 ± 0.37; P = 0.077) were counted in the first and second (ablation) MOET cycles, respectively. These preliminary findings suggest that ablation of DF before start of superovulatory treatment may be beneficial in improving the yield of transferable embryos in a buffalo MOET program. Director, CIRB, is acknowledged for the experimental facilities.


2015 ◽  
Vol 13 (3) ◽  
pp. e04SC01 ◽  
Author(s):  
Juan F. Aller ◽  
Marcos C. Abalos ◽  
Francisco A. Acuña ◽  
Rosana Virgili ◽  
Francisco Requena ◽  
...  

<p>The objectives of this study were to compare the effects of two intravaginal devices (ID) containing the same dose (0.5 g) of progesterone (P<sub>4</sub>) on subsequent ovarian response, embryo production and circulating P<sub>4</sub> concentration profile in llamas (<em>Lama glama</em>) treated with equine chorionic gonadotropin (eCG) for ovarian superstimulation. Female llamas were randomly assigned (n = 10 llamas per group) to one of the following groups and treated (Day 0) with an ID containing 0.5 g of vegetal P<sub>4</sub> to synchronize the emergence of a new follicular wave: i) DIB 0.5<sup>®</sup> and ii) Cronipres M15<sup>®</sup>. On Day 3 llamas were intramuscularly treated with 1000 IU of eCG. The IDs were removed on Day 7. Llamas were naturally mated (Day 9) and treated with GnRH analogue to induce ovulation. A second mating was allowed 24 h later. Embryos were collected between 7 and 8 days after the first mating. Blood samples were taken every day from Day 0 to Day 7 to measure circulating P<sub>4 </sub>concentrations. The results indicated that DIB device maintained greater plasma P<sub>4</sub> levels as compared to Cronipres until Day 2. However, the mean (± SD) number of corpora lutea and recovered embryos was not affected (<em>p </em>&lt; 0.05) by the type of ID (5.3 ± 2.6 <em>vs</em> 4.2 ± 2.2 and 3.5 ± 2.7 <em>vs</em> 2.6 ± 3.0 for DIB and Cronipres, respectively). In conclusion, both DIB and Cronipres devices can be successfully used to synchronize the emergence of follicular wave prior to a single dose of eCG in superovulation protocol in llamas.</p>


2011 ◽  
Vol 23 (3) ◽  
pp. 438 ◽  
Author(s):  
J. A. Skidmore ◽  
M. Billah

The aim of the present study was to investigate the use of exogenous progesterone and equine chorionic gonadotrophin (eCG) in non-ovulated and ovulated, asynchronous dromedary camel recipients being prepared for an embryo transfer programme. The uteri of 12 mated donor camels were flushed non-surgically 7 days after ovulation and 42 embryos were recovered. In Experiment 1, 16 embryos were transferred non-surgically to recipients on Day 3 or 4 after ovulation (ov+3 and ov+4, respectively). Each recipient received a daily dose of 75 mg, i.m., progesterone-in-oil from 2 days before embryo transfer until 6 days after ovulation. Thereafter, the progesterone dose was reduced to 50 mg on Day 7 and finally to 25 mg day–1 on Days 8 and 9. Nine of 16 recipients (56%; ov+3, n = 4; ov+4, n = 5) became pregnant compared with none of eight non-progesterone treated controls, into which embryos were transferred on Day 4 after ovulation. In Experiment 2, 18 non-ovulated recipients received 75 mg, i.m., progesterone-in-oil daily from 3 days before until 12 days after non-surgical transfer of a Day 7 blastocyst, at which time pregnancy was diagnosed by ultrasonography. All pregnant recipients continued to receive 75 mg progesterone-in-oil daily for a further 6 days, when each camel received 2000 IU, i.m., eCG. Progesterone treatment was then reduced to 50 mg day–1 and, when a follicle(s) ≥1.3 cm in diameter were present in the ovaries, each animal received 20 μg buserelin to induce ovulation. Once the corpora lutea had developed, progesterone treatment was reduced to 25 mg day–1 for a final 3 days. Fourteen of 18 recipients (78%) became pregnant and seven of these (50%) remained pregnant after eCG treatment. Of the seven pregnancies that were lost, two were lost before eCG treatment, two did not respond to eCG treatment and three responded to eCG treatment and ovulated, but lost their pregnancies 6–8 days after the last progesterone injection.


1972 ◽  
Vol 55 (3) ◽  
pp. 599-607 ◽  
Author(s):  
B. T. DONOVAN ◽  
A. N. LOCKHART

SUMMARY The release of ovulating hormone after acute treatment with gonadal steroids, or corpus luteum removal on different days of the oestrous cycle, was studied in the guinea-pig. Injection of 25, 50 or 100 μg oestradiol or 2·5 mg progesterone on day 13 of the cycle had no effect upon gonadotrophin secretion as judged by follicular histology, but markedly altered the sizes of the corpora lutea of the previous ovulation. Treatment with oestradiol on day 14 did not elicit gonadotrophin secretion. However, administration of the same hormones to animals given 10 μg oestradiol benzoate 24 h earlier caused ovulation or follicular luteinization. Progesterone (2·5 mg) appeared least effective in stimulating gonadotrophin release; 25 μg oestradiol were more effective when given at 12.00 h than at 24.00 h but treatment with both hormones caused ovulation when given at either time of day. Luteal volumes were not affected. Removal of corpora lutea during the second half of the cycle advanced the time of expected ovulation to day 15 or earlier when the procedure was carried out on days 8 or 9, but not on days 10–13. It is concluded that 4–5 days must elapse between the fall in plasma progesterone level associated with corpus luteum regression and the release of ovulating hormone.


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