Synchronization of estrus and ovulation in sows not conceiving in a scheduled fixed-time insemination program

A field study was conducted to investigate the effectiveness of a treatment with altrenogest, eCG and hCG or the GnRH-analogue D-Phe(6)-LHRH to synchronize estrus and ovulation of sows diagnosed as non-pregnant in order to reintegrate them back into a scheduled fixed-time insemination program. Sows (n=531) diagnosed as non-pregnant by ultrasonography on days 21-35 after insemination were subjected to one of three treatments: (1) 16 mg altrenogest/day/animal orally for 15 days to block follicular growth, followed by injection of 1000 IU eCG intramuscularly (i.m.) 24h after withdrawal of altrenogest to stimulate follicular growth and 500 IU hCG i.m. 78-80 h after eCG to induce ovulation; (2) similar to (1) except that 20mg altrenogest and 800 IU eCG were used and (3) similar to (2) except that 50 microg D-Phe(6)-LHRH was used to induce ovulation. Females were artificially inseminated (AI) twice at 24 and 40 h, respectively, after hCG/D-Phe(6)-LHRH. Success of treatments was checked by ultrasonography of the ovaries. Rates of conception and farrowing (CR, FR), and number of total and live born piglets (TB, LB) were recorded and compared to those of synchronized first served sows. Females had differing ovarian structures prior to treatment. Altrenogest effectively blocked follicular growth in >80% of the females irrespective of dosage, but 16 mg increased the development of polycystic ovarian degeneration. Four to 18% of the females still had corpora lutea after altrenogest. Most females ovulated either between both inseminations or thereafter (P<0.05). Females treated with D-Phe(6)-LHRH tended to ovulate earlier than those injected with hCG. The CR and FR were up to 25% lower for sows diagnosed as non-pregnant than for sows after first service (P<0.05). Among sows diagnosed as non-pregnant the CR was higher in females treated with D-Phe(6)-LHRH (P<0.05). No differences were found in regard to numbers of TB and LB. In conclusion, a treatment with 20mg altrenogest per day per animal, followed by 800 IU eCG and 50 microg the GnRH-analogue D-Phe(6)-LHRH is appropriate to synchronize estrus and ovulation of sows diagnosed as non-pregnant. Whether there might be a need to feed altrenogest for a longer interval of 18 days has to be investigated.     

Synchronization of ovulation in cyclic gilts with porcine luteinizing hormone (pLH) and its effects on reproductive function

The overall objective was to evaluate the use of porcine luteinizing hormone (pLH) for synchronization of ovulation in cyclic gilts and its effect on reproductive function. In an initial study, four littermate pairs of cyclic gilts were given altrenogest (15 mg/d for 14 d). Gilts received 500 microg cloprostenol (Day 15), 600 IU equine chorionic gonadotropin (eCG) (Day 16) and either 5mg pLH or saline (Control) 80 h after eCG. Blood samples were collected every 4h, from 8h before pLH/saline treatment to the end of estrus. Following estrus detection, transcutaneous real-time ultrasonography and AI, all gilts were slaughtered 6d after the estimated time of ovulation. Peak plasma pLH concentrations (during the LH surge), as well as the amplitude of the LH surge, were greater in pLH-treated gilts than in the control (P=0.01). However, there were no significant differences between treatments in the timing and duration of estrus, or the timing of ovulation within the estrous period. In a second study, 45 cyclic gilts received altrenogest for 14-18d, 600 IU eCG (24h after last altrenogest), and 5mg pLH, 750 IU human chorionic gonadotropin (hCG), or saline, 80 h after eCG. For gilts given pLH or hCG, the diameter of the largest follicle before the onset of ovulation (mean+/-S.E.M.; 8.1+/-0.2 and 8.1+/-0.2mm, respectively) was smaller than in control gilts (8.6+/-0.2mm, P=0.05). The pLH and hCG groups ovulated sooner after treatment compared to the saline-treated group (43.2+/-2.5, 47.6+/-2.5 and 59.5+/-2.5h, respectively; P<0.01), with the most synchronous ovulation (P<0.01) in pLH-treated gilts. Embryo quality (total cell counts and embryo diameter) was not significantly different among groups. In conclusion, pLH reliably synchronized ovulation in cyclic gilts without significantly affecting embryo quality.     

Evaluation of systems for collection of porcine zygotes for DNA microinjection and transfer

Crossbred gilts and sows (n=116) were used for the collection of 1-cell zygotes for DNA microinjection and transfer. Retrospectively, estrus synchronization and superovulation schemes were evaluated to assess practicality for zygote collection. Four synchronization and superovulation procedures were used: 1) sows were observed for natural estrous behavior; 1000 IU human chorionic gonadotrophin (hCG) was administered at the onset of estrus (NAT); 2) cyclic gilts were synchronized with 17.6 mg altrenogest (ALT)/day for 15 to 19 days followed by superovulation with 1500 IU pregnant mares serum gonadotropin (PMSG) and 500 IU hCG (LALT); 3) gilts between 11 and 16 days of the estrous cycle received 17.6 mg ALT for 5 to 9 days and PMSG and hCG were used to induce superovulation (SALT); and 4) precocious ovulation was induced in prepubertal gilts with PMSG and hCG (PRE). A total of 505 DNA microinjected embryos transferred into 17 recipients produced 7 litters and 50 piglets, of which 8 were transgenic. The NAT sows had less (P < 0.05) ovarian activity than gilts synchronized and superovulated by all the other procedures. Synchronization treatments with PMSG did not differ (P > 0.05) in the number of corpora hemorrhagica or unovulated follicles, but SALT and PRE treaments had higher ovulation rates than LALT (24.7 +/- 2.9, 24.3 +/- 1.8 vs 11.6 +/- 2.7 ovulations; X +/- SEM). The SALT and PRE treatments yielded 12.3 +/- 2.6 and 17.7 +/- 1.7 zygotes. Successful transgenesis was accomplished with SALT and PRE procedures for estrus synchronization and superovulation.     

In vitro development of embryos from superovulated gilts treated with the progesterone agonist, altrenogest (Regu-Mate) or the prostaglandin analogue, cloprostenol (Planate)

This study was conducted to compare the in vitro development of embryos from superovulated postpubertal gilts synchronized with progesterone agonist altrenogest (REG, Regu-Mate) and those from superovulated prepubertal gilts synchronized with prostaglandin analogue cloprostenol (PLA, Planate). Ten postpubertal gilts that had exhibited estrus at least once were fed 20 mg/d of REG from Day 0 (the first day of treatment, may have been any day of the estrous cycle) to Day 17. The gilts received intramuscularly (im) 1500 IU of equine chorionic gonadotropin (eCG) on the afternoon of Day 17, followed by 1000 IU of human chorionic gonadotropin (hCG) 84 h later. Eight prepubertal gilts received intramuscularly one dose of a combination of 400 IU of eCG and 200 IU of hCG (PG 600) on Day 0 (the first day of treatment), followed by 750 IU of hCG on Day 3. From Day 16 to Day 19, the prepubertal gilts received 350 mg/d of PLA, followed by 1500 IU of eCG on the afternoon of Day 19, then 1000 IU of hCG 84 h later. Gilts were checked for estrus with an intact boar. At estrus, all gilts were artificially inseminated and/or mated twice at 12-h intervals. Then 50 to 54 h after the hCG injection, a mid-ventral laparotomy was performed on each gilt. Corpora albicans (CA) and corpora hemorrhagica (CH) were counted, and oviducts were flushed in situ. The embryos recovered (1- to 2-cell) were cultured in modified Whitten's medium at 38.5 degrees C under an atmosphere of 5% CO2 in air for 144 h. The number of CA per gilt did not differ between the postpubertal and prepubertal gilts (11.9 vs 7.9, respectively; P > 0.05). However, the number of CH per gilt (27.5 vs 18.1, P = 0.05) and the number of embryos per gilt (26.2 vs 15.3, P < 0.05) were higher in postpubertal gilts than in prepubertal gilts. Furthermore, after 144 h of in vitro culture, the percentage of embryos cleaving to the >-16-cell (morula + blastocysts) or > or =32-cell (blastocysts) was greater (P < 0.05) in prepubertal gilts than in postpubertal gilts (85.2 vs 68.5, 55.7 vs 44.2, respectively). The total numbers of embryos examined were 122 and 260 in prepubertal and postpubertal gilts, respectively. These results show that postpubertal gilts treated with REG produced a higher number of embryos. However, better embryo development was noted with zygotes from prepubertal gilts primed with exogenous gonadotrophin, followed by synchronization with prostaglandin before induction of superovulation and insemination.     

Effect of insemination of estrus-induced prepuberal gilts on ensuing reproductive performance and body weight

Prepuberal gilts reared and managed to 85-90 kg live weight in a common system were allocated at random to one of three first-mating treatments in an experiment conducted over a period of more than 5 years. In two of the treatments, gilts received a single i.m. injection of 400 IU equine chorionic gonadotropin (eCG) and 200 IU human chorionic gonadotropin (hCG) (PG600; Intervet) and were either inseminated 4 and 5 days later on a fixed-time basis regardless of oestrus (treatment A), or at the second oestrus following treatment (treatment B). The third group of gilts remained untreated and was inseminated on the first spontaneous oestrus (treatment C). Thereafter, all gilts were managed in the same way and those observed in oestrus were re-inseminated. Significantly more gilts returned to oestrus after the first service in treatment A (35%) than in treatment B and C (12 and 17%, respectively; P<0.01). Gilts farrowed to the first or repeat inseminations at a significantly younger age (P<0.01) in treatment A (304 days) than treatment B (324 days) and C (320 days). The age difference at farrowing remained in surviving gilts at the end of their third parity. The first farrowing performance of the gilts was significantly affected by treatment in terms of litter size at birth (A 7.0, B 8.4 and C 8.3 live piglets per gilt; P<0.01), litter size at weaning (A 6.2, B 7.2 and C 7.2 live piglets per gilt, P<0.05), and piglet birth weight (A 1.4, B 1.3 and C 1.3 kg; P<0.05) but piglet survival rate and weaning weight were not affected by treatment. The live weights of the gilts were significantly different between the treatments at first insemination (A 95.7, B 106.5 and C 109.2 kg; P<0.01) but not when the first litter was weaned (A 133.6, B 135.1 and C 136.6; P>0.05). After the first farrowing there were no differences between the treatments in terms of the survival rate, productive or reproductive performance of the gilts/sows and their offspring. Without conducting a detailed cost-benefit-calculation it was deduced that, from an economical point of view, differences between treatment A and treatments B and C are negligible because the savings associated with farrowing at a younger age on this treatment just about compensated for any additional costs associated with the treatment and the lower number of piglets born at the first farrowing.     

Effect of superovulation induction on embryonic development on day 5 and subsequent development and survival after nonsurgical embryo transfer in pigs

To evaluate the effects of eCG dosage on recovery and quality of Day 5 embryos and on subsequent development and survival after embryo transfer, batches of 5 to 10 donor sows were treated with 1000 or 1500 IU eCG. Recipients from the same batch were synchronously treated with 800 IU eCG. Ovulation was induced with 750 IU hCG (72 h after eCG) in donors and recipients. Donors were inseminated and embryos were collected at 162 h after hCG (120 h after ovulation). Ovulation rate was lower using 1000 IU eCG (28.5+/-11.7; n=48) than 1500 IU eCG (45.7+/-20.3; n=32; P<0.0001). Embryo recovery rate (82.9+/-16.9%) and percentage expanded blastocysts (56.2+/-31.4%) were similar (P>0.05). Expanded blastocysts from each group of sows were pooled into 2 groups within eCG treatment, containing embryos from normally ovulating sows (< or = 25 corpora lutea [CL]) or from superovulated sows (> 25 CL). Average diameter and number of cells of a random sample of the expanded blastocysts per pool were recorded. The average diameter of blastocysts (160.5+/-11.5 microm) was not affected by eCG dosage or ovulation rate (P>0.10). The average number of cells per embryo was higher in the 1000 IU eCG group (84.3+/-15.3) than in the 1500 IU eCG group (70.2+/-1.9; P<0.05) but was similar for normal and superovulated donors within each eCG group (P>0.10). Of the 4 groups, litters of 28 to 30 blastocysts were nonsurgically transferred to 27 synchronous recipients. Pregnant recipients were slaughtered on Day 37 after hCG treatment to evaluate embryonic development and survival. Pregnancy rate for the 1000 and 1500 IU eCG donor groups was 71% (10/14) and 46% (6/13; P>0.10), respectively. The number of implantations and fetuses for the 1000 IU eCG groups was 12.9+/-3.0 and 11.1+/-2.7, and 14.2+/-7.0 and 10.5+/-4.6, respectively, for the 1500 IU eCG groups (P>0.10). After post-priory categorizing the litters of blastocysts to below or above the average diameter (158 microm) of the transferred embryos, irrespective of eCG dosage or ovulation rate, the pregnancy rate was 43% (6/14) and 77% (10/13; P<0.10), respectively. Post-priory categorizing the transferred litters to below or above the average number of cells per embryo litter, irrespective of eCG dosage or ovulation rate, showed no differences in pregnancy rates or number of implantations and fetuses (P>0.10). It was concluded that eCG dosage affects embryonic development at Day 7 after hCG, and this effect was not due to ovulation rate. Embryonic survival after nonsurgical transfer was not related to eCG dosage but tended to be related to the diameter of the blastocysts.     

Fixed-time artificial insemination protocols on brazilian locally adapted breed gilts on ovulatory response and embryo production

The aim of this study was to use estrus synchronization protocols to favor fixed-time artificial insemination and consequently fixed-time embryo collection, and increase embryo production using eCG, in gits. In a cross over design, nine Piau breed gilts were subjected to 18 days of oral progesterone; P4 group did not receive any further; GnRH group received 25µg of GnRH 104 hours after the final application of P4; and eCG+GnRH group received 1000IU of eCG 24 hours after the final P4 in addition to GnRH for subsequent embryo collection, that was performed six days after first AI, by laparotomy. Artificial insemination was performed after 12 and 24 hours of estrus in P4 group, and 128 and 144 hours in GnRH and eCG+GnRH groups. The number of CL (8.6±3.9; 8.3±2.1; 26.7±15.0) and anovulatory follicles (4.3±3.7; 3.9±3.9; 17.2±9.5) was higher in the eCG+GnRH gilts (P<0.05). However, the use of 1000 IU of eCG reduced (P<0.05) the number of total structures (5.2±3.6; 5.1±3.1; 1.7±2.7), viable embryos (5.0±3.5; 4.8±3.3; 0.4±0.7), freezable embryos (3.6±3.4; 3.3±3.8; 0.1±0.3) and recovery rate (63.7±38.9; 58.6±24.7; 5.38±9.5). P4 and GnRH protocols were effective in the production and recovery of embryos. However, the use of 1000 IU of eCG, 24 hours after P4, was not effective in promoting the production of embryos, although the animals had superovulated.     

Induction and synchronization of ovulations of nulliparous and multiparous sows with an injection of gonadotropin-releasing hormone agonist (Receptal)

The objective of this study was to determine if administration of a set dose (10 μg) of a gonadotropin-releasing hormone agonist, buserelin (Receptal; Rc), at set times after altrenogest (Regumate; RU) treatment or after weaning was able to induce and synchronize ovulation in female swine (gilts and sows). The pubertal (n = 187) gilts were allocated to four groups, all synchronized with RU. Group 1 (RU) was inseminated twice at detected estrus, Group 2 (RU+Rc120) and Group 4 (RU+Rc104) received 10 μg Rc at 120 or 104 h after the end of RU treatment, respectively, and Group 3 (RU+eCG+Rc104) was treated with 800 IU equine chorionic gonadotropin (eCG) at 24 h and Rc 104 h after the end of RU treatment, respectively. Gilts were inseminated twice at predetermined times, namely 144 and 168 h (Group 2), 128 and 144 h (Group 3), and 144 and 152 h (Group 4) after the end of RU treatment, respectively. Pregnant gilts were slaughtered at 30 d. Administration of Rc 104 h after the end of RU feeding synchronized ovulation over a 24-h time window in 97.9% and 100% of the gilts of Groups 3 and 4, respectively, whereas Rc administration at 120 h (Group 2) only successfully synchronized 88.9% of the gilts over 24 h. Ovulation rates of gilts of Groups 2 and 4 were similar to that of the control group. Pregnancy rates were numerically higher in Groups 2 and 3 (92% and 96%, respectively) compared with those of Groups 1 and 4 (84% and 81%, respectively). Combination of eCG with Rc administration at 104 h (Group 3) increased ovulation rate (+4 CL) but decreased embryo survival to 62% at Day 30. The weaned sow experiment involved 61 sows of a range of parities (2.7 ± 0.9), allocated to two control groups (Control 104 group and Control 94 group) and two treated groups (Rc104 group and Rc94 group), which received 10 μg Rc at 104 and 94 h after weaning, respectively. The females were inseminated at detected estrus. All pregnant sows farrowed. After treatment with Rc 94 h after weaning, 100% of sows ovulated over a 24-h time window versus only 68.7% of controls. Farrowing rate and litter size of the sows treated with Rc at that time were unaffected compared with that of control sows. In contrast, Rc administration at 104 h after weaning may have been too late; only 66.7% of the treated sows ovulated during a 24-h period. This proportion was numerically lower but not significantly different than that for control sows. Farrowing rate and litter size of treated sows were not significantly different than that of controls. Administration of Rc at the dose and times selected in this study tightened synchrony of ovulation in gilts and in sows after weaning. It remains to be established if such a synchrony is suitable to obtain good fertility after a single artificial insemination at a predetermined time.     

Suppression of ovarian activity in the gilt and reversal by exogenous gonadotrophin administration

The aim of the current experiment was to study the regulation of follicle development in the pig using a potent GnRH agonist (GnRH-A) to initially suppress follicle development. Large-White hybrid gilts (n = 8) were treated during the luteal phase with GnRH-A. Four of these GnRH-A treated gilts and four control gilts were given a GnRH bolus on days 14 and 28 after GnRH-A administration or during the luteal phase in control gilts. Blood samples were collected for 10 h for FSH and LH, after which 1500 IU PMSG were administered and the ovaries and uteri recovered 72 h later. A further four GnRH-A treated gilts and four control gilts were slaughtered either 28 days after GnRH-A administration or during the luteal phase respectively, and all follicles > or = 1 mm diameter were dissected. The mean basal plasma FSH level was lower (P < 0.01) in GnRH-A treated than control gilts and showed no response to the GnRH challenge although levels increased (P < 0.01) in control gilts. The mean basal plasma LH levels were similar (P > 0.1) in GnRH-A treated and control gilts. Whilst in GnRH-A treated gilts plasma LH levels showed no response to the GnRH challenge, plasma LH levels were increased (P < 0.01) in control gilts. Pulsatile LH secretion was abolished in GnRH-A treated but not in control gilts. Plasma oestradiol levels were lower (P < 0.001) in GnRH-A treated gilts than in control gilts, but nevertheless both GnRH-A treated and control gilts responded to PMSG with increased plasma oestradiol levels. Treatment with GnRH-A reduced both the mean (2.1 vs. 2.7 mm; P < 0.01) and the maximal follicle diameter (4 vs. 6 mm) and reduced (P < 0.01) the total number of follicles > or = 2 mm diameter compared with control gilts. Administration of PMSG increased both mean follicle diameter (5.1 vs. 4.4 mm; P < 0.01) and maximal follicle diameter (7 vs. 9 mm) and caused a reduction (P < 0.001) in the total number of follicles > or = 2 mm diameter in both GnRH-A treated and control gilts. In summary, this study has demonstrated, for the first time in the pig, that the inhibition of follicle development as a result of pituitary down regulation/desensitisation can be reversed by exogenous gonadotrophin treatment. This model will be a powerful tool with which to investigate the precise regulation of follicle development in the pig.     

Seminal plasma does not advance ovulation in hCG-treated sows

In gilts, seminal plasma treatment before or during the LH-surge has been found to advance ovulation in all animals by as much as 8 to 14 h. Two experiments were performed to assess whether such an advancement occurs in multiparous sows in which ovulation is induced by 750 i.u. hCG at 68 h after weaning. In both experiments, seminal plasma was inseminated at 4, 5 and 6 h after hCG (7 and 6 sows, respectively) and control sows (6 and 6 sows, respectively) were not inseminated. In Experiment 1, using Meishan semen, all sows ovulated between 38 and 44 h after hCG; no advancement of ovulation was seen due to treatment. In Experiment 2, using GY seminal plasma, 3 and 4 sows, respectively had started ovulation at 44 h after hCG. Again, no advancement of ovulation was seen due to treatment. Therefore, in both experiments, seminal plasma treatment within 4–6 h after hCG failed to advance ovulation to a similar extent as found in spontaneously ovulating gilts. It is unclear what causes this lack of effect. Maybe seminal plasma treatment does not advance hCG-induced ovulation or batches of seminal plasma differ in their ovulation-advancing properties.     

Effect of eCG dose and ovulation induction treatments on embryo recovery and in vitro development post-vitrification in two selected lines of rabbit does

The aim of this work was to evaluate the effect of different doses of eCG administered subcutaneously (0, 50 and 200 IU) and the hormonal induction of ovulation (GnRH or hCG) on embryo recovery and in vitro development of embryos post-vitrification in two selected lines of rabbit does. The two selected lines were line V (selected for the litter size at weaning) and line R (selected for growth rate). Administration of 200 IU of eCG significantly increased ovulation rate (19.2 +/- 1.2 versus 15.5 +/- 1.1 and 12.2 +/- 1.3, and the number of haemorrhagic follicles (13.8+/-1.6 versus 3.8+ /- 1.4 and 3.8 +/- 1.7), but significantly decreased recovery rate (28.8 +/- 6.3 versus 47.7 +/- 5.7 and 48.7 +/- 6.7, 200 IU versus 50 IU and 0 IU eCG, respectively), the number of normal embryos recovered per doe with at least one embryo (5.8 +/- 0.9 versus 8.2 +/- 0.9, 200 IU versus 50 IU eCG doses) and the in vitro development of embryos post-vitrification (51.9% versus 66.1%, 200 IU versus 50 IU eCG doses, respectively). Inducing ovulation with hCG significantly increased ovulation rate when compared with GnRH (17.3 +/- 0.8 versus 13.8+/-1.4), but no significant differences in embryo recovery and embryo development post-vitrification were observed between the two treatments. No significant differences were observed between the two selected lines in ovulation and recovery rates, the number of haemorrhagic follicles and the number of recovered embryos per doe. However, the post-vitrification in vitro rate of development was 59.7% for line R and 51.9% for line V (p < 0.05). It was concluded that the use of 50 IU of eCG subcutaneous with hCG or GnRH prior to embryo cryopreservation programmes in rabbits achieves the best results for embryo recovery, with the best development of recovered embryos post-vitrification.     

Fixed-time deep uterine insemination in PGF2α-synchronized goats

The aim of this investigation was to optimize fixed-time insemination in goats by clustering ovulations in prostaglandin F_2α-synchronized goats either with gonadotropin releasing hormone (GnRH) or human chorionic gonadotropin (hCG). The underlying intention was to reduce the incidence of short cycles by providing a more sustained stimulation of the corpus luteum by substituting the commonly used GnRH with longer-acting hCG. It was conjectured that this might render the corpus luteum less prone to premature regression. Sixty pluriparous does were administered 5 mg of the prostaglandin F_2α preparation dinoprost (Dinolytic; Pharmacia and Upjohn, Erlangen, Germany) during the luteal phase of the estrous cycle. Twenty of these does were administered 0.004 mg of the GnRH analog buserelin (Receptal; Intervet, Unterschleissheim, Germany) 48 hours later; another 20 does received 500 IU hCG (Chorulon; Intervet, Unterschleissheim, Germany) instead. Sixteen hours later the does were inseminated with frozen-thawed semen. The remaining 20 does served as controls and were inseminated 16-18 h after the onset of detected estrus. All 60 treated goats displayed estrous symptoms, the time of onset being similar for all groups (42.6, 37.6, and 40.5 hours after treatment for GnRH-treated, hCG-treated, and control does, respectively). The duration of estrus in the GnRH-treated group was 10 h less than in the other groups (45.1 vs. 56.4 and 54.4 h, P < 0.05). The number of ovulations (assessed by ultrasound monitoring) did not differ among groups (2.4, 2.1, and 2.5, P > 0.05). Monitoring of serum progesterone revealed that the incidence of corpus luteum insufficiency was significantly higher in GnRH- and hCG-treated does than in the control group (40% and 35% vs. 5%, P < 0.05). The pregnancy rate was 50% in the GnRH and 35% in the hCG group as compared with 60% in the controls. Corresponding kidding rates were 40%, 35%, and 60% (P > 0.05). When disregarding does with corpus luteum insufficiency, pregnancy rates would have been 83%, 54%, and 63%, and kidding rates 67%, 54%, and 63%, respectively. The average number of kids born was 1.88, 1.71, and 1.83, respectively (P > 0.05). It may be concluded that fixed time insemination of cycling does treated with prostaglandin F_2α during the luteal phase, followed by ovulation induction with GnRH or hCG, would be an effective management tool if it were possible to control the high incidence of corpus luteum insufficiency. The attempt to achieve this by substituting GnRH with hCG, was not met with success. Until a solution for the problem has been found, it is advisable to inseminate prostaglandin-synchronized does 16-18 hours after the onset of detected estrus.     

Fixed-time deep uterine insemination in PGF2α-synchronized goats

The aim of this investigation was to optimize fixed-time insemination in goats by clustering ovulations in prostaglandin F_2α-synchronized goats either with gonadotropin releasing hormone (GnRH) or human chorionic gonadotropin (hCG). The underlying intention was to reduce the incidence of short cycles by providing a more sustained stimulation of the corpus luteum by substituting the commonly used GnRH with longer-acting hCG. It was conjectured that this might render the corpus luteum less prone to premature regression. Sixty pluriparous does were administered 5 mg of the prostaglandin F_2α preparation dinoprost (Dinolytic; Pharmacia and Upjohn, Erlangen, Germany) during the luteal phase of the estrous cycle. Twenty of these does were administered 0.004 mg of the GnRH analog buserelin (Receptal; Intervet, Unterschleissheim, Germany) 48 hours later; another 20 does received 500 IU hCG (Chorulon; Intervet, Unterschleissheim, Germany) instead. Sixteen hours later the does were inseminated with frozen-thawed semen. The remaining 20 does served as controls and were inseminated 16–18 h after the onset of detected estrus. All 60 treated goats displayed estrous symptoms, the time of onset being similar for all groups (42.6, 37.6, and 40.5 hours after treatment for GnRH-treated, hCG-treated, and control does, respectively). The duration of estrus in the GnRH-treated group was 10 h less than in the other groups (45.1 vs. 56.4 and 54.4 h, P < 0.05). The number of ovulations (assessed by ultrasound monitoring) did not differ among groups (2.4, 2.1, and 2.5, P > 0.05). Monitoring of serum progesterone revealed that the incidence of corpus luteum insufficiency was significantly higher in GnRH- and hCG-treated does than in the control group (40% and 35% vs. 5%, P < 0.05). The pregnancy rate was 50% in the GnRH and 35% in the hCG group as compared with 60% in the controls. Corresponding kidding rates were 40%, 35%, and 60% (P > 0.05). When disregarding does with corpus luteum insufficiency, pregnancy rates would have been 83%, 54%, and 63%, and kidding rates 67%, 54%, and 63%, respectively. The average number of kids born was 1.88, 1.71, and 1.83, respectively (P > 0.05). It may be concluded that fixed time insemination of cycling does treated with prostaglandin F_2α during the luteal phase, followed by ovulation induction with GnRH or hCG, would be an effective management tool if it were possible to control the high incidence of corpus luteum insufficiency. The attempt to achieve this by substituting GnRH with hCG, was not met with success. Until a solution for the problem has been found, it is advisable to inseminate prostaglandin-synchronized does 16–18 hours after the onset of detected estrus.     

Effect of a GnRH analogue (Maprelin) on the reproductive performance of gilts and sows

The ability of peforelin (l-GnRH-III) to stimulate follicular growth, FSH release, and estrus in gilts after altrenogest treatment and in sows after weaning was investigated. In three farrow-to-wean herds, with at least 600 sows and average production performance, 216 gilts, 335 primiparous, and 1299 pluriparous sows were randomly allocated to three treatments: peforelin (M group: Maprelin), eCG (F group: Folligon), and physiological saline solution (C group). Animals were treated 48 hours after their last altrenogest treatment (gilts) or 24 hours after weaning (sows). The weaning-to-estrus interval, estrus duration, estrus rate (ER), pregnancy rate, and total born (TB), live born, and stillborn (SB) numbers were recorded and compared between treatments for the different parity groups (gilts and primiparous and pluriparous sows). Follicle sizes were measured in representative animals from each group on the occasion of their last altrenogest treatment or at weaning, and also on the occasions of their first (FS1) and second (FS2) attempted inseminations. Blood samples were taken to determine FSH concentrations at weaning and 2 hours after injection, and progesterone concentrations 10 days after the first insemination attempt. The relative change in FSH concentrations was calculated. Significant differences were found for ER within 7 days of weaning in pluriparous sows (95%, 91%, and 90% for the M, F, and C groups, respectively, P = 0.005). Gilts in the F-group had high TB numbers, and pluriparous sows in the M group had high SB numbers (TB gilts = 13.6, 15.4, and 14.9 [P = 0.02] and SB pluriparous sows = 1.8, 1.4, and 1.7 [P = 0.05] for the M, F, and C groups, respectively). The M group had the highest FS1 (for gilts) and FS2 (for pluriparous sows) values: FS1 = 5.4, 4.9, and 4.9 mm [P = 0.02] and FS2 = 6.8, 5.3, and 6.3 mm [P = 0.03] for the M, F, and C groups, respectively. There were no significant differences between the different treatments within each parity group with respect to any of the other variables. Overall, peforelin treatment had small but positive effects on the ER and follicle growth in certain parity groups but did not seem to affect litter sizes or FSH and progesterone levels in sows on the occasions of the corresponding examinations.     

The effects of administration of gonadotrophins and estrogens on prenatal survival in gilts

In gilts ovulation occurs over a 4 to 8-hour period, with 70% of the ova being shed over a relatively short span of time. These oocytes supposedly give rise to more developed embryos at Days 10 to 12 which advance the uterine environment and reduce survival rates of less developed embryos because of an asynchronous environment. The aim of this experiment was to reduce embryo mortality by influencing the duration and pattern of ovulation. Crossbred gilts (n = 98) were bred at their first observed estrus after being exposed to boars at 200 days of age. Estrus detection was carried out daily at 0000, 0800 and 1600 hours. All gilts were artifically inseminated with fresh semen, with a minimum of 2.7 billion spermatozoa, at both 16 and 32 hours after detection of estrus. Gilts were randomly assigned to one of the following treatments at detection of estrus: 1) 500 IU (2ml) chorionic gonadotrophin (hCG) injected intravenously at the onset of estrus (n = 22); 2) 16 mug (4 ml) gonadotrophin releasing hormone (GnRH) injected intravenously at the onset of estrus (n = 25); 3) 11.5 mug estrogen added to the semen at the time of AI (n = 25); 4) control, untreated gilts (n = 26). All gilts were slaughtered at Day 30 of gestation (Day 0 = day of detected estrus). The mean (+/-SEM) number of ovulations in pregnant gilts per treatment was 13.0 +/- 0.52, 12.6+/-0.51, 13.6+/-0.54 and 13.3+/-0.52, while the mean (+/-SEM) number of normal embryos per treatment was 10.3+/-0.67, 10.5+/-0.66, 10.3 +/- 0.69 and 10.5 +/- 0.67 for hCG, GnRH, estrogen and control groups, respectively, for an embryonic survival rate of 80 +/- 4.2%, 83 +/- 4.1%, 74 +/- 4.3% and 79+/-4.2% in pregnant gilts. If nonpregnant gilts are included, the embryonic survival rate for treatments 1 to 4 was 76+/-7.0%, 73+/-6.5%, 60+/-6.5%, and 64+/-6.4%, respectively. There was no significant difference between treatments for any of these variables. There was no evidence that administration of hCG, or GnRH at the onset of estrus, or the addition of estrogen to semen improved embryonic survival in gilts by Day 30 in this experiment.     

The effects of superovulation of donor sows on ovarian response and embryo development after nonsurgical deep-uterine embryo transfer

This study aimed to evaluate the effectiveness of superovulation protocols in improving the efficiency of embryo donors for porcine nonsurgical deep-uterine (NsDU) embryo transfer (ET) programs. After weaning (24 hours), purebred Duroc sows (2–6 parity) were treated with 1000 IU (n = 27) or 1500 IU (n = 27) of eCG. Only sows with clear signs of estrus 4 to 72 hours after eCG administration were treated with 750 IU hCG at the onset of estrus. Nonhormonally treated postweaning estrus sows (n = 36) were used as a control. Sows were inseminated and subjected to laparotomy on Days 5 to 6 (Day 0 = onset of estrus). Three sows (11.1%) treated with the highest dosage of eCG presented with polycystic ovaries without signs of ovulation. The remaining sows from nonsuperovulated and superovulated groups were all pregnant, with no differences in fertilization rates among groups. The number of CLs and viable embryos was higher (P < 0.05) in the superovulated groups compared with the controls and increased (P < 0.05) with increasing doses of eCG. There were no differences among groups in the number of oocytes and/or degenerated embryos. The number of transferable embryos (morulae and unhatched blastocysts) obtained in pregnant sows was higher (P < 0.05) in the superovulated groups than in the control group. In all groups, there was a significant correlation between the number of CLs and the number of viable and transferable embryos, but the number of CLs and the number of oocytes and/or degenerated embryos were not correlated. A total of 46 NsDU ETs were performed in nonhormonally treated recipient sows, with embryos (30 embryos per transfer) recovered from the 1000-IU eCG, 1500-IU eCG, and control groups. In total, pregnancy and farrowing rates were 75.1% and 73.2%, respectively, with a litter size of 9.4 ± 0.6 piglets born, of which 8.8 ± 0.5 were born alive. There were no differences for any of the reproductive parameters evaluated among groups. In conclusion, our results demonstrated the efficiency of eCG superovulation treatments in decreasing the donor-to-recipient ratio. Compared with nonsuperovulated sows, the number of transferable embryos was increased in superovulated sows without affecting their quality and in vivo capacity to develop to term after transfer. The results from this study also demonstrate the effectiveness of the NsDU ET procedure used, making possible the commercial use of ET technology by the pig industry.     

The use of GnRH or estradiol to facilitate fixed-time insemination in an MGA-based synchronization regimen in beef cattle

Two experiments were conducted to compare pregnancy rates when GnRH or estradiol were given to synchronize ovarian follicular wave emergence and ovulation in an MGA-based estrus synchronization program. Crossbred beef cattle were fed melengestrol acetate (MGA, 0.5 mg per day) for 7 days (designated days 0-6, without regard to stage of the estrous cycle) and given cloprostenol (PGF; 500 microg intramuscular (im)) on day 7. In Experiment 1, lactating beef cows (n=140) and pubertal heifers (n=40) were randomly allocated to three groups to receive 100 microg gonadorelin (GnRH), 5 mg estradiol-17beta and 100 mg progesterone (E+P) in canola oil or no treatment (control) on day 0. All cattle were observed for estrus every 12 h from 36 to 96 h after PGF. Cattle in the GnRH group that were detected in estrus 36 or 48 h after PGF were inseminated 12 h later; the remainder were given 100 microg GnRH im 72 h after PGF and concurrently inseminated. Cattle in the E+P group were randomly assigned to receive either 0.5 or 1.0 mg estradiol benzoate (EB) in 2 ml canola oil im 24 h after PGF and were inseminated 30 h later. Cattle in the control group were inseminated 12 h after the first detection of estrus; if not in estrus by 72 h after PGF, they were given 100 microg GnRH im and concurrently inseminated. In the absence of significant differences, all data for heifers and for cows were combined and the 0.5 and 1.0 mg EB groups were combined into a single estradiol group. Estrus rates were 57.6, 57.4 and 60.0% for the GnRH, E+P and control groups, respectively (P=0.95). The mean (+/-S.D.) interval from PGF treatment to estrus was shorter (P<0.001) and less variable (P<0.001) in the E+P group (49.0+/-6.1 h) than in either the GnRH (64.2+/-15.9 h) or control (66.3+/-13.3 h) groups. Overall pregnancy rates were higher (P<0.005) in the GnRH (57.6%) and E+P (55.7%) groups than in the control group (30.0%) as were pregnancy rates to fixed-time AI (47.5, 55.7 and 28.3%, respectively). In Experiment 2, 122 crossbred beef heifers were given either 100 microg GnRH or 2 mg EB and 50 mg progesterone in oil on day 0 and subsequently received either 100 microg GnRH 36 h after PGF and inseminated 14 h later or 1 mg EB im 24 h after PGF and inseminated 28 h later in a 2 x 2 factorial design. Pregnancy rates were not significantly different among groups (41.9, 32.2, 33.3 and 36.7% in GnRH/GnRH, GnRH/EB, EB/GnRH and EB/EB groups, respectively). In conclusion, GnRH or estradiol given to synchronize ovarian follicular wave emergence and ovulation in an MGA-based synchronization regimen resulted in acceptable pregnancy rates to fixed-time insemination.     

Ovarian response, embryo recovery and results of embryo transfer in a Hungarian native pig breed

The objective of the study was to use embryo transfer (ET) for propagation of the Swallow Belly Mangalica population. Mangalica is a native Hungarian pig breed adapted to extreme climate and housing conditions and distinguished for excellent meat and fat quality. However, due to their weak reproductive characteristics and relatively high fat proportion, Mangalica pigs have been replaced by modern breeds. Now, there is an increased interest again to safeguard the properties of this breed. We conducted two experiments. First, we used a total of 18 puberal Mangalica gilts to determine an optimal superovulatory treatment. Following estrus synchronization with Regumate, we injected gilts with either 750, 1000 or 1250 IU PMSG, followed by 750 IU hCG 80 h later. We scanned ovaries endoscopically 3 days after hCG administration. The application of 1000 and 1250 IU PMSG resulted in a higher rate of ovulation compared to 750 IU (24.2 +/- 3.6 and 21.0 +/- 2.3 vs. 13.7 +/- 2.7 P<0.05). The number of follicular cysts increased after administration of 1250 IU PMSG compared to 750 and 1000 IU (2.0 +/- 1.3 vs. 0.3 +/- 0.7 and 0.2 +/- 0.3, P<0.05). Thus, we chose 1000 IU PMSG for further stimulation of Mangalica gilts. In the second experiment, we induced superovulation in 10 Mangalica donor gilts by 1000 IU PMSG and 750 IU hCG. Gilts were fixed-time inseminated, and then five days later embryo collection was carried out surgically (n=6) or endoscopically (n=4). Out of the 187 ova recovered, 92.5% were at the morula/blastocyst stage. The embryo recovery rate was higher following surgical flushing than following endoscopy (91.5 +/- 4.4% vs. 71.4 +/- 12.7%, P<0.05). Altogether 143 embryos were transferred surgically or endoscopically into 8 Landrace recipients. Surgical and endoscopic transfer of Mangalica embryos into Landrace gilts resulted in pregnancies in 3 and 2 gilts, respectively; thus the overall farrowing rate was 62.5%. The birth of 59 Mangalica piglets from 5 embryo recipients equals an average litter size of 11.8 +/- 1.3, which is two times larger than usual in this breed. Therefore, we concluded that an appropriate inter-breed ET program is a suitable tool to propagate the endangered Mangalica breed.     

Ovulation and pregnancy outcomes in response to human chorionic gonadotropin before resynchronized ovulation in dairy cattle

We first determined a dose of human chorionic gonadotropin (hCG) sufficient to induce ovulation in lactating Holstein cows. Ovaries of 85 previously inseminated cows were mapped using transrectal ultrasonography 7 d before pregnancy diagnosis and assigned randomly to treatments of saline, 100 μg gonadotropin-releasing hormone (GnRH), or 500, 1000, 2000, or 3000 IU hCG. Appearance of new corpus luteum (CL) in response to ≥1000 IU hCG was similar to that for GnRH but greater (P < 0.001) than that for saline. Ovarian structures and serum progesterone then were monitored in 334 previously inseminated Holstein cows 0 and 7 d after treatment with GnRH, hCG (1000 IU), or saline. The incidence of ovulation was greater (P = 0.01) after GnRH than after saline in cows having pretreatment progesterone < 1 ng/mL, whereas in cows having progesterone ≥1 ng/mL, GnRH or hCG was more (P = 0.01) effective than saline, and hCG also differed from GnRH. Holstein cows of unknown pregnancy status in three herds were treated with either GnRH, hCG, or as controls to initiate an ovulation-resynchronization procedure 7 d before pregnancy diagnosis. In 1109 treated pregnant cows, pregnancy loss during 4 wk after treatment tended (P = 0.06) to be greater in those treated with hCG. Treated cows (n = 1343) diagnosed not pregnant were then given prostaglandin F_2α and inseminated and received GnRH 72 h later. A treatment by herd interaction (P = 0.06) resulted in more pregnancies after GnRH in two herds and after hCG in one herd compared with saline. We concluded that (1) ≥ 1000 IU hCG resulted in more CL than did treatment with saline, and the incidence of new CL after either GnRH or hCG depended on pretreatment progesterone status; (2) hCG tended to increase pregnancy loss in pregnant cows; and (3) pregnancies per artificial insemination after initiating resynchronization with either hCG or GnRH produced ambiguous results.