Use of fluorogestone acetate after breeding to reduce the effect of premature luteal regression in dairy goats when superovulation is induced with FSH

The aim of the present study was to determine the effect of fluorogestone acetate (FGA) administered after mating, on embryo production in the dairy goat subjected to conventional superovulatory and embryo recovery protocols. Adult does, most of them of the French Alpine breed, were randomly assigned after a FSH-superovulatory estrus and fertile matings to a control group (n=20) or to a treated group (n=20) in which intravaginal sponges impregnated with FGA were inserted after mating and remove before embryo collection (day 6). Blood samples were collected every 12h from days 1 to 7 post-estrus and serum progesterone concentrations were determined. The FGA-group had a lesser percentage of does with normal corpora lutea (CL) and a greater percentage of animals with CL in regression or mixed (normal and in regression) when compared with the control group (13.3 and 64.7%, 53.3 and 23.5%, and 33.3 and 11.8%, respectively; P<0.05). Mean number of normal CL per doe was less and mean number of regressed CL greater in FGA as compared with the control group (4.2 compared with 10.7 and 8.5 compared with 3.6, respectively; P<0.05). There were no differences (P>0.05) in recovery rate, total number of CL, total recovered structures, oocytes and transferable and non-transferable embryos between groups. Serum progesterone concentrations from day 5 to 7 post-estrus were lower (P<0.05) in FGA as compared with the control group. Percentage of does with luteal failure on day 6 post-estrus was greater in FGA as compared with the control group (86.6 compared with 33.3%; P<0.01). When considering only does with luteal failure on day 6 post-estrus, mean total recovered structures, transferable embryos and percentage of does rendering > or =3 transferable embryos were greater in the FGA compared with the control group (6.3 and 1.3 structures, 4.5 and 1.2 embryos, 67 and 17%, respectively; P<0.05). In does not having luteal failure, FGA administration did not appear to affect embryo production or embryo survival. These results indicate that FGA administration after mating improves embryo recovery in dairy goats with luteal failure after superovulatory treatment. However, it also increases the incidence of luteal regression when administered early in the estrous cycle.     

Ovsynch synchronization and fixed-time insemination in goats

This study assessed the efficacy of an Ovsynch protocol (vs. the classical cronolone containing vaginal sponge+eCG treatment) to generate fixed-time insemination in goats during the breeding season. Each regimen was applied to 24 Boer goat does. Onset and duration of estrus were determined with an aproned male and follicular development was monitored by ultrasonography. Ovulation and quality of the corpora lutea were established from progesterone concentrations. In 10-11 goats per group, LH concentrations were determined throughout the preovulatory period. Does were inseminated at pre-determined times (16 h after the second GnRH injection and 43 h after sponge removal). Estrus was identified in 96% of the Ovsynch-treated goats (at 49 h after prostaglandin injection) and in 100% of the goats synchronized with sponges (at 37 h after sponge removal). Low progesterone concentrations at the time of AI were observed in 21/24 and 24/24 goats synchronized by Ovsynch and sponges, respectively. Synchronization of the LH surge was tighter following Ovsynch compared to sponge treatment. Kidding rates (at 58 and 46% in the Ovsynch and sponge groups, respectively) and prolificacy (at 1.86 and 1.83 in the Ovsynch- and sponge-treated goats) were similar for both groups, as were the number of ovulations (2.9 and 3.3) and the proportion of does with premature corpus luteum regression (29 and 17%). When excluding does with premature luteal regression and those with low progesterone levels when receiving prostaglandins, kidding rate reached 87.5% (14/16) after Ovsynch. During the breeding season, the Ovsynch protocol may thus be an useful alternative to the sponge-eCG treatment.     

Good quality sheep embryos produced by superovulation treatment without the use of progesterone devices

Multiple ovulation and embryo transfer (MOET) is a very important tool for the genetic improvement and preservation of endangered livestock. However, the success of a MOET programme highly depends on the number of transferable embryos in response to a superovulation treatment. Thus, the aim of this study was to compare the number and quality of embryos produced during natural oestrus under porcine FSH treatment without the use of progesterone devices to more traditional protocols. Forty Sarda sheep were divided into 2 groups: without sponges (WS) (n = 20) and with sponges (S) containing 40mg FGA for 12 d (n = 20) (control group); 350 I.U. of porcine FSH per sheep was administered in eight decreasing doses twice daily starting four days after estrus was detected (Day 0) in group WS and 48 h before sponge removal in group S. A single i.m. dose of 125 μg of cloprostenol was administered on Day 6 after estrus in group WS to induce luteolysis. Sheep were naturally mated 24 h after cloprostenol injection or sponge removal. Seven days after mating, an inguinal laparotomy was performed and the number of corpora lutea (CL) recorded. Embryos were recovered surgically by flushing each uterine horn. A total of 38 fresh and 22 vitrified embryos were transferred in pairs into 3 groups of recipients seven days after estrus detection: fresh embryos from group S (S-F) (n = 9), fresh embryos from group WS (WS-F) (n = 10) and vitrified embryos from group WS (WS-V) (n = 11). Data on the number of corpora lutea (CL), recovered ova and embryos (OER), and quality 1-2 and 3 embryos (EQ_1-2, EQ₃) per ewe were analyzed by ANOVA. Recovery (RR), fertility (FR) and quality 1-2 embryo (Q_1-2R) rates per treatment were analyzed by a Chi Square analysis. A Chi Square analysis was also applied to pregnancy rate (PR), lambing rate (LR) and twinning rate (TR) of fresh and vitrified embryos in order to analyze embryo transfer results. Among all superovulation variables analysed, results show statistically significant differences in mean number of CL/ ewe (9.3 ± 3.9 vs 7 ± 3.2), RR (67% vs 80 %) and FR (100% vs 80%) (P < 0.05) between WS and S groups respectively. There were no significant differences in PR (78%, 70% and 82%), LR (67%, 60% and 59%) and TR (71%, 71% and 44.4%) among S-F, WS-F and WS-V groups respectively. In conclusion, it is possible to produce a good number of transferable embryos during natural oestrus avoiding the use of sponges.     

Highly synchronous and fertile reproductive activity induced by the male effect during deep anoestrus in lactating goats subjected to treatment with artificially long days followed by a natural photoperiod

The response to the male effect was studied in two flocks of Saanen and three of Alpine goats during deep anoestrus in three consecutive years. Males and females were subjected to artificially long days for about 3 months (between December 4 and April 1) followed by a natural photoperiod. Bucks joined goats 42-63 days after the end of the long days treatment (between April 20 and June 3) and fertilisation was ensured by natural mating. In experiment 1 (n=248), female goats were treated or untreated with melatonin at the end of the long days treatment and treated or untreated for 11 days with fluorogestone acetate (FGA) before teasing. The males received melatonin implants. In experiment 2 (n=337), the factor studied was the association or non-association of the 11-day FGA treatment. Neither males nor females received melatonin implants. In experiment 3 (n=180), goats were treated for 11 days with FGA or with natural progesterone (CIDR). Neither males nor females received melatonin implants. In experiment 1, among the non-cycling goats (n=218), 99% ovulated and 81% kidded at 161+/-8 days after joining. Ninety-two percent of FGA-treated goats displayed an LH surge at 65+/-11h after teasing. Melatonin treatment did not affect any parameter but FGA advanced the kidding date. In experiment 2, 94% of the goats ovulated and 87% kidded. A major peak of conception was observed on days 3 and 8 after joining in FGA-treated and untreated goats, respectively. Among the FGA-treated goats, 83% displayed an LH surge. Over all flocks, most of the LH surges occurred over a 24-36 h interval, but the surge was initiated at different times in different flocks (36, 48 or 60 h after joining). FGA treatment did not influence the results, except for advancement of births of about 5 days. Differences among flocks were highly significant. In experiment 3, 94% of the goats displayed the LH surge, 93% ovulated and 68% kidded. Significant differences were found among flocks, but not between the FGA and CIDR groups. Bucks marked 85% of the goats 24-72 h after joining. The time interval between the detection of marked goats and detection of the LH surge depended on the time of marking (r=-0.62; p<0.05). In conclusion, treatment of both males and females goats with artificially long days followed by a natural photoperiod is very effective in inducing highly synchronous and fertile reproductive activity via the male effect in the middle of seasonal anoestrus.     

High fertility using artificial insemination during deep anoestrus after induction and synchronisation of ovulatory activity by the "male effect" in lactating goats subjected to treatment with artificial long days and progestagens

The response to the male effect was studied in two Saanen and two Alpine flocks over 5 consecutive years. Adult male and female goats were exposed to artificial long days (16h light and 8h darkness, 16L:8D) in open barns for approximately 3 months (between December 1 and April 15) followed by a natural photoperiod. Goats were treated for 11 days with fluorogestone acetate (FGA) or progesterone (CIDR) immediately before joining. Bucks carrying marking harnesses with adapted aprons joined females 49-63 days after the end of the long-day treatment (between April 30 and June 5) and were left with them for 5 days. In experiment 1 (n=142), FGA- and CIDR-treated goats were inseminated at a time based on the detection of oestrus. Two insemination groups were distinguished by the occurrence of marking over a 48-h period. Earlier (group 1) and later (group 2) buck-marked goats received one single insemination 12-24h or 0-12h after marking, respectively. Unmarked goats were inseminated along with group 2. In experiment 2 (n=344), FGA-treated goats were inseminated 52 and 70 h (52 h:70 h group) or 52 and 75 h (52 h:75 h group) after joining. In experiment 3 (n=285), FGA-treated goats were inseminated 52 h (1-AI group) or 52 and 75 h (2-AI group) after joining. In all experiments, an external control group given the "classical" insemination program was analysed. Over the 5-year period, 92% of the goats exhibited an LH surge during days 1-4 after joining and 98% of them ovulated. Eighty-seven percent of the LH surges detected in milk occurred during the 33-57 h interval after joining, indicating that ovulation took place around 45-69 h. In experiment 1, 96% of the goats were marked 22-70 h after joining. Kidding rate (KR; 78%) was similar between insemination groups and between FGA- and CIDR-treated goats (p>0.05). Most of the goats (95%) were inseminated during the interval between 15h before and up to 4h after ovulation. KR was not affected by the time between detection of marking and insemination or between insemination and ovulation (p>0.05). In experiment 2, KR (75%) was similar in both insemination groups (p>0.05). In experiment 3, KR was higher (p<0.05) in the 1-AI (71%) than the 2-AI group (57%). In all experiments, KR of the control group (68-73%) was similar to that achieved in goats induced to ovulate by the male effect. Prolificity (2.1+/-0.7) was not affected by any of the factors examined (p>0.05). In conclusion, high fertility can be achieved during anoestrus when 1 or 2 inseminations are performed over a 24h period, determined by oestrus or by the introduction of the buck, if light-treated goats receive 11-day FGA or CIDR treatment and are then induced to ovulate by the male effect.     

Premature luteal regression in goats superovulated with PMSG: effect of hCG or GnRH administration during the early luteal phase

Twenty-two goats were superovulated with PMSG; 84 h after the onset of estrus the goats were treated with saline solution (control group n = 7), hCG (hCG group, n = 7), or GnRH (GnRH group, n = 8). The ovaries of all the goats were laparoscopically examined 3 and 6 d after the onset of estrus. In each case the CL were counted and classified according to their appearance as normal-looking or as regressing. Blood samples for progesterone determination were collected every 12 h from Day 1 to Day 6. Premature luteal regression was considered to have occurred if progesterone concentrations declined to less than 1 ng/mL by Day 6. According to progesterone concentrations, 57.5, 0 and 37.5% of the goats underwent premature luteal regression in the control, hCG and GnRH groups, respectively. Progesterone concentrations were higher in the hCG group than in the other groups on Days 5 and 6 post estrus (P < 0.05). The control group was the only one in which there was a significant (P < 0.05) increase in the number of regressing CL between Day 3 (1.6 +/- 1.4) and Day 6 (7.3 +/- 1.4). It was also the only group in which there was a significant decrease in the number of normal-looking CL between Day 3 (12.6 +/- 2.1) and Day 6 (2.6 +/- 2.1). On Day 6 the animals treated with hCG had significantly more normal-looking CL (12.0 +/- 2.3) than those in the control group (2.6 +/- 2.1). The number of large follicles present on the ovaries on Day 6 post estrus had negative correlations with progesterone concentrations (P = 0.05) and with the number of normal-looking CL (P < 0.05). It is concluded that the administration of hCG 84 h after the onset of estrus prevents premature luteal regression in goats superovulated with PMSG.     

Endocrine responses of goats after induction of superovulation with PMSG and FSH

Goats in Group A were pretreated for 9 days with a synthetic progestagen, administered via intravaginal sponge, and 1000 i.u. PMSG s.c. on Day 12 of the oestrous cycle. Goats in Group B had the same PMSG treatment, but not the progestagen pretreatment. Group C goats received a s.c. twice daily injection of a porcine FSH preparation (8 mg on Day 12, 4 mg Day 13, 2 mg Day 14 and 1 mg Day 15). Oestrus was synchronized in all animals by 50 micrograms cloprostenol, 2 days after the start of gonadotrophin treatment. The vaginal progestagen sponges were removed from Group A at the same time. Mean ovulation rate was slightly higher in FSH-treated than in the PMSG-treated animals, whereas the incidence of large follicles that failed to ovulate was significantly elevated in PMSG-treated animals in Group B. More goats in Groups A and B than in Group C exhibited premature luteal failure. Progestagen pretreatment appeared to suppress both follicular and luteal activity, as indicated by numbers of large non-ovulating follicles and by the magnitude and duration of elevated plasma oestradiol levels following PMSG stimulation, and by decreased plasma progesterone levels before and after PMSG treatment. Oestrogenic response to FSH was considerably less than that to PMSG, as indicated both by a considerably shorter duration of elevation of circulating oestradiol levels during the peri-ovulatory period, and by lower maximal oestradiol levels. Differences in the ovarian responses to PMSG and FSH may be attributed primarily to differences in the biological half-life of each preparation.     

Induction/synchronization of oestrus and ovulation in dairy goats with different short term treatments and fixed time intrauterine or exocervical insemination system

Two experiments were carried out on Ionica dairy goats in order to test the efficiency of: (1) short term-5-day combined progestogen-PGF2α-GnRH treatments on induction/synchronization of oestrus and fertility after natural mating in lactating goats and during the transition period (Experiment 1); (2) short term-9-day FGA-PGF2α-eCG treatments on synchronizing oestrus and ovulation (Experiment 2.1) and artificial insemination (AI) fixed time system in synchronized does (Experiment 2.2), during the breeding season. In Experiment 1, four treatment groups (N=24) were considered: (1) FPe-11d - control, FGA intravaginal sponges (11 days)+PGF2α (9th d)+eCG (11th d); (2) FPe-5d, FGA (5 days)+PGF2α (5th d)+eCG (5th d); (3) PFe-5d, PGF2α (D0)+FGA (5 days)+eCG (5th d); (4) GPe-5d, GnRH (D0)+PGF2α (5th d)+eCG (5th d). Goats were checked for oestrus and naturally mated. The occurrence of oestrus was 75.0, 78.3, 86.4, and 58.3% for groups 1-4, respectively, with significant differences (P<0.05) between groups 3 and 4. Interval to oestrus was earlier (P<0.05) in GPE-5d than in FPe-11d control group. There were no differences between the groups (P>0.05) in fertility or in prolificacy. In Experiment 2.1, 22 goats were subdivided into two treatment groups (N=11): (T1) FPe-11d (control), FGA (11 days)+PGF2α (9th d)+eCG (11th d); (T2) FPe-9d, FGA (9 days)+PGF2α (7th d)+eCG (9th d). Oestrus and ovulation times were monitored every 4h; ovulation rate was also determined. The induction of oestrus ranged from 91 to 100% and all goats ovulated. Intervals to oestrus, from the onset of oestrus to ovulation, from sponge removal to ovulation, and ovulation rates were 28.2±4.9 and 26.0±4.0h, 25.3±9.2 and 28.9±7.4h, 53.5±7.6 and 54.9±7.1h, 3.7±1.6 and 2.4±1.4 corpora lutea (P<0.05) for T1 and T2, respectively. In T2 a great abnormal ovulatory response was observed. In Experiment 2.2, 48 goats were synchronized with FPe-9d treatment and subjected to AI, performed 50h after s.r. with frozen semen, and subdivided into 2 AI system groups (N=24): T3, exocervical AI (100×10(6)Spz/doe); T4, intrauterine AI (20×10(6)Spz/doe). Fertility rate was higher (P<0.05) in T4. It seems that short term-5-day combined progestogen-PGF2α-GnRH-eCG treatments need to be investigated for AI fixed time.     

The effect of vitamin E treatment during preovulatory period on reproductive performance of goats following estrous synchronization using intravaginal sponges

The objectives of this study were to investigate whether the use of intravaginal sponge for estrous synchronization of goats causes oxidative stress, and to examine the effect of administering vitamin E during preovulatory period on reproductive performance of estrous synchronized goats. Estrus was synchronized in 36 non-lactating adult does using intravaginal sponges containing 30 mg of fluorogestane acetate (FGA) for 14 days. All females received 500 IU of eCG at the sponge withdrawal. The goats were allocated at random to two groups balanced for breed, age and body weight. Treatment group (n=18) received 200mg of vitamin E i.m. at the time of sponge removal and again at the time of second artificial insemination. The other 18 goats (control) were administered 1 ml of physiological saline instead of vitamin E on each of these two occasions. All does in estrus was intracervically inseminated at 12 and 24h after the onset of estrus. Blood samples were collected every 72h during the experimental period for evaluation of malondialdehyde (MDA) and vitamin E concentrations. Serum MDA level increased and vitamin E concentration decreased during the period of vaginal sponge application. Following the sponge removal, MDA level declined rapidly to below basal level in the treatment group but remained high in the control group. Conversely, vitamin E concentration increased in the treatment group after the sponge withdrawal and remained at a low level in the control group. No statistically significant differences (P>0.05) were observed between groups in terms of estrous response, conception rate, gestation length or kidding rate. However, the number of multiple births (70.0% versus 50.0%) and prolificacy rate (2.40+/-0.37 versus 1.63+/-0.26 kids per kidding) were significantly higher (P<0.05) for the treatment group than those of the control group. The results indicate that the use of intravaginal sponges for estrous synchronization of goats causes an increase in level of oxidative stress. However, the vitamin E treatment during preovulatory period can prevent the overproduction of reactive oxygen species (ROS), and it may improve the multiple birth rates and the number of kids born in estrous synchronized goats.     

Embryo losses in Rasa Aragonesa ewes actively immunized against androstenedione or passively immunized against testosterone

Two-day-old embryos from untreated ewes were transferred to the oviducts of ewes actively immunized against androstenedione (n=26, Group A), passively immunized against testosterone (n=19, Group B) or left untreated (n=25, Group C). Donor ewes superovulated after treatment with follicle-stimulating hormone and fluorogestone acetate (FGA). Recipient ewes were treated with FGA and pregnant mare serum gonadotropin (PMSG, 300 I.U.). Group A received two injections of Fecundin at a 4-wk interval. FGA sponges were inserted when the second injection was given. Group B was treated with antitestosterone antiserum (35 ml) at sponge withdrawal. Each recipient received two morphologically viable embryos 52 to 62 h after the onset of estrus. Antibody titre at embryo transfer and progesterone concentration on Days 2, 4, 6, and 12 after estrus were determined. Fertility was lower in Group A when compared to Group C (42.3 vs 84.1%; P<0.01) while that of Group B (63.2%) did not differ from those of Groups A and C. In immunized groups, most of the embryo losses occurring were complete (both embryos were lost), resulting in a decreased fertility, while in the untreated group embryo losses were mainly partial (only one embryo was lost), hence lowering prolificacy. Fertility in immunized groups changed according to the antibody titre reached. Ewes from Groups A and B with higher antibody titres displayed lower fertility than control ewes. On Days 4 and 12 of the cycle, Group A plasma progesterone concentrations positively correlated with antibody titres and were higher with respect to those of Group C (P<0.05). Progesterone levels in Group B were similar to those of Group C. These results indicate that ewes reaching higher antibody levels had more embryo losses, attributable to the adverse influences of the oviductal and/or uterine environment on embryo development.     

Effects of norgestomet-implants and fluorogestone acetate-impregnated sponges on oestrous cycle length and luteal function of ewes

Plasma progesterone profiles were used to assess luteal function and length and synchronization of oestrous cycles in ewes after insertion of subcutaneous ear implants containing Norgestomet or intravaginal sponges impregnated with fluorogestone acetate (FGA) for 12 or 14 days. Insertions were made 2, 9 or 16 days after synchronization of the oestrous cycle with FGA-sponges. An i.m. injection of 500 IU pregnant mares' serum gonadotrophin was given at the time of sponge or implant removal. Norgestomet- implants inserted 9 or 16 days after FGA-sponge treatment had no effect on luteal function but delayed the onset of a new oestrous cycle for the duration of treatment. Following withdrawal of implants, oestrus was effectively synchronized. When Norgestomet-implants were inserted 2 days after FGA-sponge treatment, luteal function was normal. At the time of implant removal, plasma progesterone levels were elevated suggesting the presence of functional corpora lutea. In contrast, insertion of FGA-sponges early in the oestrous cycle shortened the luteal phase and a new oestrous cycle was initiated within 48 h after sponge removal. These results indicate that Norgestomet- implants can artificially prolong the length of the oestrous cycle and do not affect the functional lifespan of corpora lutea in cycling ewes. However, when Norgestomet-implants are inserted early in the oestrous cycle, they are unable to cause premature regression of corpora lutea.     

Induction of oestrus in Saanen goats at early breeding season by intravaginal progesterone sponges (MAP) or by prostaglandin F(2)alpha injections. Effect on different age groups

Twenty-one maiden and 29 pluriparous milking Ankara Saanen goats received either two i.m. injections of PGF(2)alpha (n=25) or intravaginal MAP sponges (n=25) early in November at the start of the breeding season. About twice as many pluriparous goats as maiden goats exhibited estrus after either treatment (87% vs. 47%). Breeding after this induced estrus caused pregnancies in 62% of the pluriparous goats, but only in 24% of the maiden animals. Maximal concentrations of progesterone were reached 11 days after the start of the MAP treatment. Progesterone declined to basal levels two to four days after sponge withdrawal. A significant slower progesterone increase also resulting in lower maximal concentrations could be observed in maiden goats. Luteolysis was evident in all animals within 24 h after PGF(2)alpha injection. Nine goats (six maiden and three pluriparous) did not exhibit Heat after the second injection and showed only a slow increase of progesterone. It seems that noncyclic animals are less sensitive to MAP treatment than to the first PGF(2)alpha injection. Goats at the beginning of the breeding season may react after a premature interruption of corpus luteum function (after second PGF(2)alpha injection) with delayed or inadequate follicular function.     

A radioimmunoassay for fluorogestone acetate (FGA) and its application to the measurement of plasma FGA and progesterone in ewes treated with FGA-impregnated intravaginal sponges

Simultaneous concentrations of endogenous progesterone and exogenous FGA have been measured in ewes treated with FGA-impregnated intravaginal sponges at several times relative to the expected time of release of LH. First, a direct double antibody radioimmunoassay (RIA) for FGA, with good precision, sensitivity and reproducibility, was developed and validated. An oxime derivative was prepared and then conjugated to human serum albumen at the 3-position to produce the antigen. Antibodies raised in New Zealand White rabbits showed little cross-reactivity with related steroids. FGA was estimated in extracted and unextracted plasma; results were indistinguishable. Second, sponges impregnated with 40 mg FGA were inserted into 20 anoestrous crossbred ewes for 12 days; 500 i.u. pregnant mare serum gonadotrophin (PMSG) was injected at withdrawal. Similar sponges were reintroduced into four ewes at each of the intervals 1, 3, 5, and 7 days later; three ewes served as controls. Plasma concentrations of progesterone and FGA were estimated by RIA daily during treatment and at intervals of 2 h for 12 h and at 18 and 24 h after withdrawal. The plasma profiles of FGA during the two successive periods of insertion were remarkably similar. A concentration of 3.0 ng/ml (s.e.m. +/- 0.22) was attained on day 1, falling to 1.5 ng/ml (+/- 0.15) by day 4. Thereafter, the concentration was maintained at 1.1 ng/ml (+/- 0.08). Plasma progesterone concentrations were at basal levels of less than 0.2 ng/ml during the first (acyclic) period of sponge insertion. During the second (cyclic) period there was a marked difference related to the time of sponge insertion. Insertion on day 1 (before LH release) resulted in complete inhibition of luteal activity; insertion on day 3, 5 or 7 was followed by apparently normal luteal function. There was no evidence of any feedback mechanism of exogenous progestagen on endogenous progesterone and no interaction. It is concluded that a 12-day treatment is needed in cyclic ewes for full synchronization and that sponges impregnated with 40 mg FGA will maintain an effective plasma concentration of greater than 1 ng/ml to the end of this period.     

How the FSH/LH ratio and dose numbers in the p-FSH administration treatment regimen, and insemination schedule affect superovulatory response in ewes

We wished to evaluate the effects of FSH/LH ratio and number of doses of p-FSH during a superovulatory treatment on ovulation rate and embryo production (Experiment I). In Experiment II, we studied the efficacy of fertilization after various insemination schedules in superovulated donors. In Experiment I estrus was synchronized in 40 ewes (FGA, for 9 days plus PGF2alpha on Day 7) and the ewes were randomly assigned to four treatment groups as follows (n = 10 ewes each): Group A: four p-FSH doses with the FSH/LH ratio held constant (1.6); Group B: four p-FSH doses with the FSH/LH ratio decreasing (FSH/LH 1.6-1.0-0.6-0.3); Group C: eight p-FSH doses with the FSH/LH ratio held constant (1.6); Group D: eight p-FSH doses and FSH/LH ratio decreasing (1.6-1.6, 1.0-1.0, 0.6-0.6, 0.3-0.3). p-FSH administrations were performed twice daily 12 h apart. The ewes were mated at the onset of estrus and again after 12 and 24 h; then, one ram per four ewes was maintained with the ewes for two additional days. Ovarian response and embryo production were assessed on Day 7 after estrus. Experiment II. Three groups (n = 10 each) of superovulated ewes were inseminated as follows: Group M: mated at onset of estrus; Group AI: artificial insemination 30 h after onset of estrus; M + AI) mating at onset of estrus and intrauterine AI performed 30 h from estrus with fresh semen. Results of Experiment I showed that treatment (D) improved (P < 0.05) ovulatory response in comparison to Groups (C) and (A). The fertilization rate was lower (P < 0.01) in Group D) than Group (A). Also the proportion of transferable embryos was lower in Group (D) in comparison to all the other treatments (P < 0.01). Group A gave the best production of embryos (7.3/ewe; 89.0% transferable). In Experiment II, combined mating plus AI improved fertilization rate (80.3%) compared to both mating (P < 0.01) and AI (P < 0.02) alone.     

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.     

Use of Chronogest implants and Folltropin for estrus synchronization and superovulation in goats.

Six Barbari goats each were assigned randomly to treatments 1,2 or 3, comprising im injections of FSH (folltropin) at 12, 14 or 16 mg dose level respectively. Estrus was synchronized with intravaginal sponge impregnated with flugestone acetate (30 mg; chronogest) inserted for 12 days and cloprostenol (125 micrograms) im at the insertion as well as at removal of sponge. FSH treatment started 48 hr before the sponge removal as 4-day declining dose scheme. Estrus could be effectively synchronized in all goats under the study, with significant difference (P less than 0.05) in the onset of estrus between the treatment groups. All goats were administered with 750 IU hCG i.v. at estrus. Recording of ovarian response and embryo recovery was done 45 hr after the onset of estrus. The prime aim of superovulation was effectively achieved in Barbari goats with the use of chronogest implants and folltropin. There was no difference (P greater than 0.05) between the treatment groups in recovery of transferable embryos, however, 14 mg folltropin appeared to be near optimal dose. There was no adverse effect on the quality of recovered embryos with high doses of folltropin.     

Ovarian response,embryo production and hormonal profile in superovulated goats treated with insulin

The influence of insulin on ovarian response and embryo production was investigated in 30 mixed breed goats, divided randomly into three equal (n=10) groups. Goats in Group 1 (control) were superovulated using 20 IU FSH i.m. in six divided descending doses, i.e. 4/4, 3/3 and 3/3 IU at 12 h interval for three consecutive days and were not given insulin treatment. Goats in Group 2 (insulin pretreatment) were pretreated with long acting purified bovine insulin 0.2 IU/kg body weight per day s.c. on Days 7, 8 and 9 of the estrous cycle prior to initiation of superovulatory treatment as in Group 1. Animals in Group 3 (insulin cotreatment) were treated as in Group I, but in addition received long acting purified bovine insulin 0.2 IU/kg body weight per day s.c. as a cotreatment along with the first, third and fifth FSH treatments on three consecutive days. Total ovarian response (corpus luteum and unovulated large follicle (UOLF)) was significantly (P<0.05) higher in insulin pretreatment (17.90+/-3.08) than in the cotreatment (11.50+/-2.34) and control (11.90+/-1.87) groups. The number of UOLF was significantly higher (P<0.05) in the insulin pretreatment (10.2+/-1.67) than the cotreatment (4.9+/-1.14) and control (3.6+/-1.09) groups. The mean transferable quality of embryos did not differ significantly among treatments. Progesterone concentration on the day of PGF(2)alpha treatment was not different (P>0.05) between the insulin treatment groups (5.28+/-0.79; 5.30+/-0.66 ng/ml). Estradiol-17beta concentration was significantly (P<0.05) higher on the day of PGF(2)alpha treatment in both the insulin treatment groups (36.67+/-6.40; 34.33+/-4.33 pg/ml) as compared to the control group (20.00+/-2.73 pg/ml). There is ample evidence to indicate beneficial effect of insulin on folliculogenesis and steroidogenesis in superovulated goats.     

Effect of type of intravaginal progestagen treatment of estrous response and reproductive performance of ewes

A comparison was made of the relative effectiveness of sponge pessaries impregnated with 40mg flourogestone acetate (FGA) or 60mg medroxyprogesterone acetate (MAP) to induce a synchronized estrus in ewes. Ewes were treated with sponge pessaries for 14 days and 500 IU pregnant mares' serum gonadotropin was injected i.m. at the time of sponge removal. The degree and pattern of mating response of ewes were similar, irrespective of the treatment used, approximately 92% of the ewes being marked by the ram by 72h after sponge removal. No significant differences in fertility or litter size were observed between the treatment groups. Ewes treated with FGA sponges had a fertility of 53% and litter size of 2.3 after mating at the synchronized estrus. The corresponding values for ewes treated with MAP sponges were 57% and 2.1. Use of MAP sponges was associated with a 17.8% sponge loss during treatment compared with 1% sponge loss in ewes treated with FGA sponges. Such losses could compromise the use of MAP sponges by reducing their overall efficacy.     

Evidence for prostaglandin involvement in early luteal regression of the superovulated nanny goat (Capra hircus)

Feral does of various ages were treated with intravaginal progestagen sponges for 16 days to synchronize oestrus. On Day 2 before sponge removal the goats were given 1200 i.u. PMSG to induce superovulation: 6 of the goats were also injected every 12 h with flunixin meglumine, a prostaglandin (PG) synthetase inhibitor, from Day 3 to 7 of the synchronized oestrous cycle. Jugular blood samples were collected from all females into heparinized syringes at daily intervals over the 2 days before sponge removal, twice daily for the next 2 days, then at hourly intervals from 09:00 to 17:00 h for 2 days and then twice daily for a further 2 days, for measurement of plasma progesterone and the PGF metabolite 13,14-dihydro-15-keto-PGF (PGFM) by radioimmunoassay. Intermittent surges in plasma PGFM concentrations were observed in hourly samples collected from 4/4 untreated females but in only 2/6 of the inhibitor-treated females (P less than 0.05), and the peak plasma PGFM concentrations were reduced in these 2 inhibitor-treated goats compared with the control goats. The corpora lutea (CL) of the inhibitor-treated females appeared to be functional as indicated by the plasma progesterone profile and endoscopic examination of CL. In the control females, however, there was evidence of premature regression of CL. These results suggest that the premature release of PGF-2 alpha may be the cause of premature regression of CL in nanny goats induced to superovulate.     

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.