G. Baril

Current status of embryo technologies in sheep and goat

This review presents an overview of the technical bases of in vivo and in vitro embryo production in sheep and goat. The current limitations of in vivo production, such as variability of response to the hormonal treatment, fertilization failure in females showing a high ovulatory response, and the importance of premature regressed CL in the goat, are described along with possibilities for improvement. The new prospects offered by in vitro embryo production, by repeated ovum pick-up from live females and by juvenile breeding, are presented along with their limiting steps and research priorities. The recent improvements of embryo production and freezing technologies could be used for constitution of flocks without risks of disease transmission and will allow wider propagation of valuable genes in small ruminants populations in the future.

Artificial breeding of adult goats and kids induced with hormones to ovulate outside the breeding season

Abstract A total of 28 336 adult goats of the Saanen and Alpine breeds were induced to ovulate before the breeding season with either a 21 day fluorogestone acetate (FGA) treatment administered vaginally and associated with Pregnant Mare Serum Gonadotrophin (PMSG; 10 745 females) or an 11 day FGA treatment, also administered vaginally but associated with PMSG and cloprostenol (17 591 females). After the first treatment, goats were inseminated twice during the induced oestrus with either liquid semen stored for a few hours at + 4°C (n = 4505) or frozen-thawed spermatozoa stored up to 6 years at −196°C (n = 6240): the percentage of goats delivering kids was higher when frozen-thawed spermatozoa were inseminated (56.7% vs. 52.1%; P Goats inseminated twice with frozen-thawed spermatozoa after administration of the second treatment (n = 6970) were more fertile than those inseminated under the same conditions after administration of the first treatment (n = 6240): 61.1% vs. 56.7%; P a.m. to 2:00 p.m. , the fertility rate was higher than for treatments which ended from 3:00 to 9:00 p.m. : 65.7% (n = 4338) compared with 60.7% (n = 2671), respectively, P After one insemination, fertility rates for all goats increased with time from the non-breeding to the breeding season (58.7%; 61.1% and 65.9%). This was more pronounced for Saanen goats (56.0%; 61.3% and 63.7%) than for Alpine goats in which fertility increased only in the 45 day period preceding the beginning of the full breeding season (61.0%–66.4%). Attempts were also made to adapt hormonal treatments to the anatomy and physiology of the female kid. For the same reason, artificial insemination (A.I.) conditions differing from those prevailing in adult females were tested. The vaginal administration of progesterone by means of a CIDR (Controlled Internal Drug Releaser) resulted in fertility levels as high as those obtained after administration of FGA through vaginal sponges. The insemination of 0.2 ml inseminates containing 100 × 106 spermatozoa produced fertility levels above 60% and no lower than those obtained with 0.5 ml inseminates containing 150 × 106 or 200 × 106 spermatozoa.

Synchronization of estrus in goats: the relationship between time of occurrence of estrus and fertility following artificial insemination.

The fertility rate for goats following artificial insemination (AI) is usually analyzed according to herd or treatment groups. However, these general information are insufficient to allow identification of specific factors which affect this individual reproductive performance. In the present experiment 640 dairy goats were used to analyze to what extent the interval from sponge removal to estrus affects the results of AI, performed at a predetermined time following sponge removal. Estrus occurred in 98.1% of experimental animals between 24 and 72 hours after sponge removal. The fertility rate was lower for goats that came into estrus later than 30 hours after sponge removal (33.3%, n = 108 than for goats that exhibited estrus earlier (65.0%, n = 520; P<0.001). The occurrence of late estrus is not age dependent, but it increases with the number of treatments that an individual animal has previously received. These results show that the low fertility rate observed in some herds after synchronization of estrus and AI may be related to the high proportion of goats with a late occurrence of estrus, and this phenomenon increases in animals that are treated repeatedly.

Estrus synchronization in dairy goats: use of fluorogestone acetate vaginal sponges or norgestomet ear implants

The ultimate aim of any estrus synchronization method is to allow artificial insemination at a predetermined time after the end of treatment. This requires a very tight synchronization of estrus which is not observed in goats after administration of the usual fluorogestone acetate (FGA)/prostaglandin (PG) F2 alpha/equine chorionic gonadotrophin (eCG) treatment. The possibility to improve the synchronization of estrus and luteinizing hormone (LH) peak with different progestagens (FGA versus norgestomet) and routes of administration (vaginal sponge versus subcutaneous ear implant) was evaluated in two experiments where goats received one of three progestagen treatments: (1) a vaginal sponge impregnated with 45 mg of FGA, (2) a half-implant of norgestomet, or (3) a whole implant containing 3 mg of norgestomet. The progestagens were left in place for 11 days and intramuscular injections of 400 or 500 IU of eCG (according to milk yield) and 50 micrograms of an analogue of PGF2 alpha (cloprostenol) were given 48 h prior to progestagen removal. In Experiment 1, 117 cycling goats were checked for the time of onset of estrus, preovulatory LH peak and ovulation rate following estrus synchronization treatment. Goats treated with half-implants came into estrus earlier than those receiving vaginal sponges (27.8 +/- 5.0 h vs. 33.0 +/- 6.6 h, respectively; P < 0.05). No effect of progestagen priming was observed on the variability of the onset of estrus. However, the interval between the time of onset of estrus and LH peak was more variable (P < 0.05) in goats treated with half-implants. In Experiment 2, 170 non-cycling goats were monitored for the time of onset of estrus, percentage of females ovulating, fertility and prolificacy after estrus induction treatment and artificial insemination with frozen-thawed semen performed 24 h after the onset of estrus. No effect of progestagen treatment was observed either on the time or the variability of onset of estrus. The percentage of goats ovulating and overall fertility rate were higher (P < 0.05) in goats receiving vaginal sponges (98.2% and 75.0%, respectively) than those treated with half-implants (81.8% and 45.5%, respectively). However, no significant difference was observed, for the same parameters, in animals receiving implants (86.3% and 58.8%, respectively). In conclusion, estrus synchronization with a norgestomet implant or half-implant did not reduce the variability in the onset of estrus and LH peak. The fertility tended to be lower in goats treated with a whole implant and was significantly decreased in goats which received a half-implant.

Synchronization of estrus in goats: The relationship between eCG binding in plasma, time of occurrence of estrus and fertility following artificial insemination

Abstract Radioimmunoassay (RIA) was used to measure plasma eCG binding in dairy goats (n = 524) at the beginning of a progestagen/eCG treatment and 25 d after eCG administration. The eCG binding was not dependent on the age of the females but increased with the number of treatments they had previously received (3.4 % ± 4.8, n = 47 vs 9.6 % ± 13.2, n = 249; mean ± SD; P

Successful direct transfer of vitrified sheep embryos

The use of a simple cryopreservation method, adapted to direct transfer of thawed embryos may help to reduce the costs of embryo transfer in sheep and increase the use of this technique genetic improvement of this species. Two experiments were made to test a vitrification method that is easy to apply in field conditions. All embryos were collected at Day 7 of the estrous cycle of FSH-stimulated donor ewes and were assessed morphologically, washed in modified PBS and incubated for 5 min in 10% glycerol, for 5 min in 10% glycerol and 20% ethylene glycol and were transferred into the vitrification solution (25% glycerol and 25% ethylene glycol). All solutions were based on mPBS. Embryos were loaded in straws (1 cm central part, the remaining parts being filled with 0.8 M galactose in mPBS) and plunged into liquid N2 within 30 sec of contact with the vitrification solution. The straws were thawed (10 sec at 20 degrees C) and the embryos were either transferred directly or after 5 min of incubation in the content of the straw (followed by washing in PBS) into the uterus of a recipient ewe. In Trial 1, the pregnancy rates at term (72 vs. 72%) as well as the embryo survival rates (60 vs 50% respectively) were not different between fresh (n = 48 embryos) and vitrified (n = 50) embryos. In a second trial no difference was observed between vitrified embryos transferred after in vitro removal of the cryoprotectant (n = 86 embryos) or directly after thawing (n = 72) both in terms of lambing rate (67 vs. 75%, respectively) and embryo survival rate (lambs born/embryos transferred; 49 vs. 53%). This method of sheep embryo cryopreservation provided high pregnancy and embryo survival, even after direct transfer of the embryos.

Induction and synchronization of estrus in goats: The relative efficiency of one versus two fluorogestone acetate-impregnated vaginal sponges

The purpose of the experiment was to test the hypothesis that a variable and/or insufficient level of progestagen at the end of a treatment to synchronize estrus in goats could explain variability in the onset of estrus. The experiment was performed during the anestrous season on 2 herds, one of Alpine (n = 49) the other of Saanen (n = 53) dairy goats. The animals were allocated to 1 of 3 treatments: Group 1 received a vaginal sponge impregnated with 45 mg of fluorogestone acetate (FGA) on Day 0; Group 2 received a sponge on Day 0 plus a second sponge on Day 7; Group 3 received a sponge on Day 0 plus a second sponge on Day 9. The sponges were withdrawn on Day 11. All goats received 400 or 500 IU eCG and 50 mug PGF(2alpha) analog 48 h prior to sponge removal. They were inseminated with frozen-thawed semen 24 h after the onset of estrus. Among treatment groups no difference (P > 0.05) was observed for the following parameters: percentage of goats in estrus, percentage of goats ovulating, mean time and variability of onset of estrus. The fertility of Alpine goats in Group 3 was significantly decreased (P < 0.05). No effect on prolificacy was noticed. These observations show that to increase progestagen level at the end of treatment did not improve estrus synchronization. They provide further evidence that treatments with too high progestagen amounts can decrease fertility.

A new method for controlling the precise time of occurrence of the preovulatory gonadotropin surge in superovulated goats

In goats treated to induce superovulation, insemination at a predetermined time after the end of progestagen treatment leads to a low fertilization rate. To solve this problem we developed a new treatment based on the control of the occurrence of the endogenous LH peak with a GnRH antagonist (Antarelix). The first experiment was designed to determine the dose of LH required to mimic a spontaneous LH preovulatory discharge; the injection of 3 mg, i.v. of pLH induced a peak of the same amplitude and duration as the spontaneous peak. Subsequently, in the second experiment, we compared 2 doses of Antarelix (0.5 and 1 mg, sc) administered 12 h after sponge removal (9 goats/treatment group). The dose of 0.5 mg was selected for further experiments because it was effective in the inhibition of the endogenous LH peak and had no detrimental effect on the quality of embryos. In the final experiment, 48 goats received the new treatment and were inseminated (intrauterine) only once 16 h after LH injection; 41 were flushed and produced 5.3 +/- 4.5 (m +/- SD) transferable embryos. The developmental stage and the number of cells/embryo were within the range that has been reported for embryos produced with conventional treatments. In conclusion, with the described method, it is possible to inseminate goats at a predetermined time without decreasing the number of transferable embryos. This technique will encourage the development of embryo transfer within genetic programs, and it will be a valuable tool for the production of zygotes for gene transfer.

Are antibodies responsible for a decreased superovulatory response in goats which have been treated repeatedly with porcine follicle-stimulating hormone ?

Repeated administration of xenogenic gonadotropins in human or animal species may be responsible for antibody production and refractoriness. An experiment was conducted in which goats were treated with porcine FSH (p-FSH) at 6-week intervals for a period of 7 months. A sensitive radioimmunoassay (RIA) was used to detect antibodies to p-FSH in plasma samples taken at short-term intervals during a 7-month period. Antibodies appeared after the first injection, and levels increased following booster injections. A high correlation rate existed between antibody level and superovulatory response. Refractoriness in goats was associated with a high level of antibodies.

The influence of ovarian status on response to estrus synchronization treatment in dairy goats during the breeding season

The influence of ovarian status (presence of a corpora lutea and follicles) on the times of the onset of estrus, LH peak and ovulation rate at a synchronized estrus was evaluated in 73 Alpine and Saanen cyclic goats. Does were treated for 11 d with 3 mg norgestomet implants or 45mg fluorogestone acetate (FGA) sponges. They also received 400 IU of PMSG and 50 μg of a PGF2α analog on Day 9 of progestagen priming. Follicles (1 to 7 mm) and corpora lutea (CL) were counted by laparoscopy on Days 0 and 9 of progestagen treatment and 5 or 6 d after the synchronized estrus. Estrus was detected every 4 h from 16 to 60 h after the end of progestagen treatment using a vasectomized buck. The LH concentration was determined by radioimmunoassay (RIA) in blood samples collected every 4 h for 24 h beginning at the time of the onset of estrus. The number of follicles on Days 0 and 9 of progestagen treatment was not related to the time of the onset of estrus and occurrence of the LH peak or to ovulation rate. The number of CL on Day 9 influenced the time of occurrence of the LH peak but not the time of the onset of estrus. Thus, in does with 2 or 3 CL on Day 9, the LH peak occurred at 46.9 h after the end of progestagen treatment, and in does with 1 or 0 CL at 42.2 and 42.5 h, respectively, after treatment, suggesting that the number of CL at luteolysis is a factor in the variability of response after the synchronization of estrus.