Efficacy of Heatsynch protocol for induction of estrus, synchronization of ovulation and timed artificial insemination in yaks (Poephagus grunniens L.)

The objective of this study was to test the efficacy of induction of estrus, synchronization of ovulation and timed artificial insemination in anestrous yaks using the Heatsynch protocol. In Experiment 1, 10 anestrous yaks were administered an analogue of gonadotropin releasing hormone (GnRH) followed by prostaglandin (PG)F2alpha 7 days later and then estradiol cyponate (ECP) 24 h after that. Ovulation was detected by rectal palpation at 2h intervals beginning at the initial signs of estrus. Blood samples were collected at 2h intervals beginning at the time of ECP injection up to 2h after the occurrence of ovulation for the determination of LH and progesterone. All the animals responded to the Heatsynch protocol with expression of estrus and synchronization of ovulation. The mean time interval from the ECP injection to ovulation was 59.4+/-2.62 h (range 50-72 h). The interval from the LH peak to ovulation was 30.2+/-2.3 h. The high degree of synchrony in ovulation could be attributed to the synchrony in the timing of LH peaks. In Experiment 2, 10 anestrous yaks were treated with the Heatsynch protocol (as in Experiment 1) and TAI was performed at 48 and 60 h after the ECP treatment. Concurrently, 16 cycling yaks were inseminated approximately 12 h after detection of spontaneous estrus. Pregnancy rates were similar in both groups, 40% for TAI and 43.75% for yaks inseminated following spontaneous estrus (p>0.05). From this study, two conclusions can be drawn. First, the Heatsynch protocol can be successfully used to induce and synchronize estrus in anestrous yaks and, second, ovulation following the Heatsynch protocol is synchronized adequately to permit the use of fixed time AI in this species.     

Efficacy of the Ovsynch protocol for synchronization of ovulation and fixed-time artificial insemination in Murrah buffaloes (Bubalus bubalis)

Two experiments were conducted to assess the timing and synchrony of ovulation, plasma LH concentrations, and pregnancy rate in Murrah buffaloes (Bubalus bubalis) treated with the Ovsynch (GnRH-PGF(2 alpha)-GnRH) protocol. In Experiment 1, 10 non-lactating cycling buffaloes received 10 microg of a GnRH analogue i.m. (buserelin acetate) without regard to the stage of the estrous cycle (day of treatment, day 0), followed by 25mg of PGF(2 alpha) i.m. (dinoprost thromethamine) 7 days later. A second-treatment of the same GnRH analogue (10 microg, i.m.) was given 48 h after PGF(2 alpha). Ovulation was confirmed by transrectal palpation (at 2-h intervals) from the second-GnRH treatment to detection of ovulation or up to 96 h after the second-GnRH treatment. Plasma LH concentrations were determined in blood samples collected at 15-min intervals for 6h, starting at the second-GnRH treatment, and thereafter at 2-h intervals until 2h after detection of ovulation. Ovulation occurred in 9/10 buffalo (90%) 23.3+/-1.3h (mean+/-S.E.M.; range 20--32 h) after the second-GnRH treatment. Peak LH concentrations 13.5+/-3.5 ng/mL (range 3.9--40.0 ng/mL) occurred 2.1+/-0.1h (range 1.2-3.0 h) after the second-GnRH treatment. In Experiment 2, 15 lactating, cycling buffaloes were subjected to the Ovsynch protocol, with fixed-time AI 12 and 24h after the second-GnRH treatment and 75 lactating buffaloes were inseminated, approximately 12h after detection of spontaneous estrus. Pregnancy rates were 33.3% for TAI and were 30.7% for buffaloes inseminated following spontaneous estrus (P=0.84). In conclusion, the Ovsynch protocol effectively synchronized ovulation in Murrah buffaloes and resulted in conception rates (to two fixed-time inseminations) that were comparable to those achieved with a single AI after detection of spontaneous estrus.     

Strategies to optimize reproductive efficiency by regulation of ovarian function in yak (Poephagus grunniens L.)

Ovulatory response to the first GnRH of Ovsynch is the critical determinant for successful synchronization of ovulation in animals. An attempt was made in this study to design a pre-Ovsynch hormonal strategy in yaks to increase the ovulatory response to the first GnRH injection of Ovsynch so that overall synchronization rate to Ovsynch could be improved. Non-lactating cyclic yak cows (n=33) were assigned to receive either no treatment before Ovsynch (control) or 0.375 mg of PGF2alpha (PreP) followed 2 d later by 10 microg of GnRH (PreG), administered 4 (G4G), 5 (G5G), or 6 (G6G) d before initiating the Ovsynch protocol. Rectal palpation was performed to assess ovulation and blood samples were collected to measure progesterone concentrations during pre-treatment, treatment and post-treatment periods. All the animals received timed AI 12 and 24h after the final GnRH of Ovsynch. Diagnoses for pregnancy were performed by rectal palpation and profiles of plasma progesterone 35 d after AI. Percentage of yak cows that ovulated in response to the first GnRH injection of Ovsynch, synchronized to Ovsynch treatment, had a functional CL at PGF2alpha of Ovsynch, had circulating concentrations of P4 at PGF2alpha of Ovsynch and likely to be pregnant after 35 d after AI, were greater in G6G and G5G compared with control, whereas G4G did not differ from controls. In addition, animals that ovulated in response to first GnRH of Ovsynch had greater response to PGF2alpha of Ovsynch and greater synchronization rate to the overall protocol than those that did not ovulate. In summary, PGF2alpha-and-GnRH-based pre-Ovsynch strategies consisting of 5 or 6-d interval between PreG and first GnRH of Ovsynch resulted in a greater ovulatory and luteolytic response to first GnRH andPGF2alpha of Ovsynch, respectively, compared with control animals. These, in turn, optimized synchronization rate to Ovsynch in yaks.     

Induction of estrus and ovulation: why some mares respond and others do not

The two most common procedures for breeding management of mares involve induction of luteolysis and induction of ovulation. Although both of these events are usually achieved, physiologic conditions affect the timing of the response. In a diestrus mare treated with prostaglandin F(2alpha) (PGF), or a PGF analogue, it is well documented that, on average, the interval from treatment to the onset of estrus is 3-4 days, whereas ovulation occurs 8-10 days after treatment. However, the diameter of the ovulatory follicle, as well as its status at the time of PGF treatment, determines the intervals from treatment to onset of estrus and to ovulation; these intervals can range from 48h to 12 days. Ovulation is routinely induced with human chorionic gonadotropin (hCG), recombinant LH (rLH), or the GnRH analogue Deslorelin. On average, ovulation occurs approximately 36h after treatment, but the effectiveness of any of these treatments can be affected by the stage of the estrus cycle, follicle size and maturity.     

Comparison of two Ovsynch protocols (GnRH versus LH) for fixed timed insemination in buffalo (Bubalus bubalis)

We evaluated the efficiency of replacing GnRH with LH in the ovulation synchronization protocol in buffaloes. Buffaloes received GnRH on Day 0, (Buserelin; Conceptal, 20 microg), PGF2alpha (Luprostiol; Prosolvin, 15 mg) on Day 7 and GnRH (Buserelin; Conceptal, 10 microg; Group 1) or porcine LH (LH; Lutropin-V, 12.5 mg; Group 2) on Day 9. In Experiment 1, we studied the follicular dynamics of 30 buffaloes (Group 1, n = 15 and Group 2, n = 15). We performed ultrasonography every 12 h from Days 0 to 2, then on Day 7 and then every 6 h from the time of GnRH or LH treatment (Day 9) until the time of ovulation. All females not ovulating by 48 h after the second GnRH or LH injection were considered as nonresponders. In Experiment 2, we evaluated 305 buffaloes (Group 1, n = 154; Group 2, n = 151), using the same two treatments studied in Experiment 1. We also recorded and evaluated aspects like parity, lactational status, the presence of mucus, and uterine tone at the time of artificial insemination (Al). In Experiment 1, ovulation rate after the first GnRH was 86.6% (26/30). Ovulation rates were 93.3% (14/15; Group 1) after the second dose of GnRH and 93.3% (14/15) after LH (Group 2). Ovulation occurred 36.4+/-10.4 h after the first GnRH. The interval for treatment to ovulation was 26.5+/-9.6 h for buffaloes treated with GnRH (Group 1) and 24.4+/-7.9 h for buffaloes treated with LH (Group 2); the time of ovulation did not differ statistically between the two groups (GnRH versus LH; P > 0.05). In Experiment 2, conception rates of the animals AI in the field were 56.5% (Group 1) and 64.2% (Group 2), respectively (P = 0.08). The response to the treatment with LH was not different to the treatment with GnRH; however, multiparous buffaloes had higher conception rates than the primiparous buffaloes in both groups (P > 0.05). Buffaloes with mucus at the time of AI in Group 2 had higher conception rates than the buffaloes that had mucus in Group 1 (P < 0.05). Uterine tone and lactational status did not influence conception rates (P > 0.05). In summary, the results showed that both treatments resulted in synchronization of ovulation and acceptable conception rates. Therefore, the exogenous injection of LH can substitute the GnRH injections in the Ovsynch program in buffaloes.     

Luteal activity at the onset of a timed insemination protocol affects reproductive outcome in early postpartum dairy cows

This study was designed to compare two timed insemination protocols, in which progesterone, GnRH and PGF2alpha were combined, with the Ovsynch protocol in presynchronized, early postpartum dairy cows. Reproductive performance was also evaluated according to whether cows showed high or low plasma progesterone concentration, at the onset of treatment. One hundred and six early postpartum dairy cows were presynchronized with two cloprostenol treatments given 14 days apart, and then assigned to one of the three treatment groups. Treatments for the synchronization of estrus in all three groups started 7 days after the second cloprostenol injection, which was considered Day 0 of the actual treatment regime. Cows in the control group (Ovsynch, n=30) were treated with GnRH on Day 0, PGF2alpha on Day 7, and were given a second dose of GnRH 32 h later. Cows in group PRID (n=45) were fitted with a progesterone releasing intravaginal device (PRID) for 9 days, and were given GnRH at the time of PRID insertion and PGF2alpha on Day 7. In group PRID/GnRH (n=31), cows received the same treatment as in the PRID group, but were given an additional GnRH injection 36 h after PRID removal. Cows were inseminated 16-20 h after the administration of the second GnRH dose in the Ovsynch group, and 56 h after PRID removal in the PRID and PRID/GnRH groups. Ovulation rate was determined on Day 11 postinsemination by detecting the presence of a corpus luteum in the ovaries. Lactation number, milk production, body condition at the onset of treatment and treatment regime were included as potential factors influencing ovulation and pregnancy after synchronization. Logistic regression analysis for cows with high and low progesterone concentration on treatment Day 0 revealed that none of the factors included in the models, except the interaction between progesterone and treatment regime, influenced the risk of ovulation and pregnancy significantly. In cows with high progesterone concentration at treatment onset, Ovsynch treatment resulted in a significantly improved pregnancy rate over values obtained following PRID or PRID/GnRH treatment. In cows with low progesterone concentration, PRID or PRID/GnRH treatment led to markedly increased ovulation and pregnancy rates with respect to Ovsynch treatment. These findings suggest the importance of establishing ovarian status in early postpartum dairy cows before starting a timed AI protocol, in terms of luteal activity assessed by blood progesterone.     

Effect of timing of Cosynch on fertility of lactating Holstein cows after first postpartum and Resynch timed-AI services

To compare two intervals from the PGF(2alpha) injection to the second GnRH injection+timed artificial insemination (TAI) of Ovsynch, lactating Holstein cows received their first postpartum TAI after Presynch + Ovsynch (n=352) and second and greater postpartum TAI after resynchronization of ovulation using Ovsynch (Resynch; n=458). Each week, cows housed in each of four breeding pens were randomized by breeding pen to receive the second GnRH injection of Presynch + Ovsynch or Resynch and TAI either 48 h (Cosynch 48; n=382) or 72 h (Cosynch 72; n=428) after the PGF(2alpha) injection of Ovsynch or Resynch. Overall, pregnancies per AI (P/AI) did not differ for cows receiving Cosynch 48 (29%) versus Cosynch 72 (33%). Furthermore, treatment did not affect P/AI for cows receiving first postpartum TAI after Presynch + Ovsynch, for cows receiving second and greater TAI after Resynch, or the proportion of female calves born. In conclusion, delaying the second GnRH injection and TAI from 48 to 72 h after the PGF(2alpha) injection of Ovsynch did not affect P/AI or calf sex ratio. The lack of a difference in fertility between these Cosynch protocols may offer more flexibility for implementing a systematic synchronization protocol when a Cosynch strategy is used.     

Calves born from anestrus yaks (Poephagus grunniens L.) subjected to Ovsynch and superovulation treatment

An attempt was made to induce estrus and ovulation in eight anestrus yaks by use of the Ovsynch protocol. Six out of eight yaks were successfully induced into estrus, and ovulation occurred in all the responding yaks 1-2 days after the second GnRH administration. Out of the six animals that responded to the treatment, two mated naturally with yak bulls, and calves were obtained from them. The other four animals were further administered a superovulatory regimen of Folltropin (FSH-P). Following Folltropin and Ilerin (a PGF(2alpha) analog) treatment, the animals were subjected to natural insemination. Only one animal in which natural mating occurred was flushed non-surgically for embryo recovery 7 days post-insemination. Thereafter, all the donor animals were administered with Ilerin. After 48-72 h, they came into heat and mated naturally with yak bulls, and calves were obtained from them after expiration of the normal gestation period. Following superovulation, the average numbers of palpable corpora lutea in the right and left ovaries were 2.25+/-0.6 and 1.75 +/-0.3, respectively. Three embryos were recovered by non-surgical flushing from a single animal. One embryo was transferred to a recipient yak, who produced one female calf after 258 days. This is the first report of production of a yak calf through embryo transfer-technology.     

Synchronization rate, size of the ovulatory follicle, and pregnancy rate after synchronization of ovulation beginning on different days of the estrous cycle in lactating dairy cows

Recently a protocol was developed that precisely synchronizes the time of ovulation in lactating dairy cows (Ovsynch; GnRH-7d-PGF2 alpha-2d-GnRH). We evaluated whether initiation of Ovsynch on different days of the estrous cycle altered the effectiveness of this protocol. The percentage of cows (n = 156) ovulating to the first GnRH was 64% and varied (P < 0.01) by stage of estrous cycle. Treatment with PGF2 alpha was effective, with 93% of cows having low progesterone at second GnRH. The overall percentage of cows that ovulated after second GnRH (synchronization rate) was 87% and varied by response to first GnRH (92% if ovulation to first GnRH vs 79% if no ovulation; P < 0.05). There were 6% of cows that ovulated before the second injection of GnRH and 7% with no detectable ovulation by 48 h after second GnRH. Maximal diameter of the ovulatory follicle varied by stage of estrous cycle, with cows in which Ovsynch was initiated at midcycle having the smallest follicles. In addition, milk production and serum progesterone concentration on the day of PGF2 alpha affected (P < 0.05) size of the ovulatory follicle. Using these results we analyzed pregnancy rate at Days 28 and 98 after AI for cows (n = 404) in which Ovsynch was initiated on known days of the estrous cycle. Pregnancy rate was lower for cows expected to ovulate larger follicles than those expected to ovulate smaller follicles (P < 0.05; 32 vs 42%). Thus, although overall synchronization rate with Ovsynch was above 85%, there were clear differences in response according to day of protocol initiation. Cows in which Ovsynch was initiated near midcycle had smaller ovulatory follicles and greater pregnancy rates.     

Timing of ovulation in relation to onset of estrus and LH peak in yak (Poephagus grunniens L.)

The objective of the study was to determine the timing of ovulation in relation to onset of estrus and the preovulatory LH peak in yaks. For this purpose, a sensitive LH enzymeimmunoassay previously established in buffaloes was successfully validated for measuring the hormone in yak plasma. Plasma LH and progesterone were estimated from blood samples collected from eight non-lactating cycling yaks at 2 h intervals after estrus onset until 6 h after ovulation (ovulation was confirmed by palpation of ovaries per rectum). The mean+/-S.E.M. preovulatory plasma LH peak was 10.11+/-0.35 ng/ml with the values ranging from 8.75 to 11.51 ng/ml in individual yaks. The mean+/-S.E.M. duration of the LH surge was 7.25+/-0.55 h with a range of 6-10 h. Onset of LH surge (mean+/-S.E.M.) occurred 3.0+/-0.65 h after the onset of estrus. Mean plasma progesterone stayed low (<0.25 ng/ml) during the entire duration of sampling. Ovulation occurred 30.5+/-0.82 h (range, 28-34 h) after the onset of estrus and 20.25+/-1.03 h after the end of LH surge. The occurrence of the LH peaks within a narrow time frame of 4-8h post estrus onset in yaks could have contributed to the animals ovulating within a narrow time interval.     

Comparison of endocrine changes,timing of ovulations,ovarian follicular growth,and efficacy associated with Estradoublesynch and Heatsynch protocols in Murrah buffalo cows (Bubalus bubalis)

Poor estrus expression and the difficulty encountered in predicting the time of ovulation compromise the reproductive efficiency of Murrah buffalo cows. Synchronization of ovulation and timed artificial insemination are able to precisely control the time of ovulation and thus avoid the need for estrus detection. Recently, the Estradoublesynch protocol (administration of a PGF2α injection 2 days before Heatsynch protocol; GnRH 0, PGF2α 7, estradiol benzoate [EB] 8) was developed that precisely synchronized ovulation twice, i.e., after GnRH and EB injections and resulted in satisfactory pregnancy rates in Murrah buffaloes. The present study was conducted on 104 cycling and 31 anestrus buffaloes to compare (1) the endocrine changes, timing of ovulations, ovarian follicular growth, and efficacy of Estradoublesynch and Heatsynch protocols in cycling and (2) the efficacy of Estradoublesynch and Heatsynch protocols for the improvement of fertility in cycling and anestrus Murrah buffalo cows. Ovulation was confirmed after all GnRH and EB treatments by ultrasonographic examination at 2-hour intervals. Plasma progesterone and total estrogen concentrations were determined in blood samples collected at daily intervals, beginning 2 days before the onset of protocols until the day of second ovulation detection. Ovulatory follicle size was measured by ultrasonography at six time points (first PGF2α administration of Estradoublesynch protocol every 2 days before the onset of Heatsynch protocol, GnRH administration of both protocols, 2 hours before ovulation detection after GnRH administration of both protocols, second PGF2α injection of Estradoublesynch protocol, PGF2α injection of Heatsynch protocol, EB injection of both protocols and, 2 hours before ovulation detection after EB administration of both protocols). Plasma LH, total estrogen, and progesterone concentrations were determined in blood samples collected at 30-minute intervals for 8 hours, beginning GnRH and EB injections, and thereafter at 2-hour intervals until 2 hours after the detection of ovulation. The first ovulatory rate was significantly higher (P < 0.05) in the Estradoublesynch protocol (84.6%) than that in the Heatsynch protocol (36.4%). The first LH peak concentration (74.6 ± 10.4 ng/mL) in the Estradoublesynch protocol was significantly higher (P < 0.05) than that of the Heatsynch protocol (55.3 ± 7.4 ng/mL). In Estradoublesynch protocol, the total estrogen concentration gradually increased from the day of GnRH administration coinciding with LH peak, and then gradually declined to the basal level until the time of ovulation detection. However, in Heatsynch protocol, the gradual increase in total estrogen concentration after GnRH administration was observed only in those buffalo cows, which responded to treatment with ovulation. In both Estradoublesynch and Heatsynch protocols, ovulatory follicle size increased by treatment with GnRH and EB until the detection of ovulation. The pregnancy rate after the Estradoublesynch protocol (60.0%) was significantly higher (P < 0.05) than that achieved after the Heatsynch protocol (32.5%). Satisfactory success rate using the Estradoublesynch protocol was attributed to the higher release of LH after treatment with GnRH, leading to ovulation in most of the animals and hence creating the optimum follicular size at EB injection for ovulation and pregnancy to occur.     

Fertility following fixed-time AI or insemination at observed estrus in Ovsynch and Heatsynch programs in lactating dairy cows

The objective of this study was to compare the conception rate for fixed-timed artificial insemination (FTAI) and observed heat artificial insemination (HAI) prior to the scheduled FTAI in Ovsynch and Heatsynch synchronization protocols. In Experiment 1, lactating dairy cows (n=535) received two set-up injections of 25mg prostaglandin F(2alpha) (PGF(2alpha)) i.m., 14 days apart starting at 36+/-3 days in milk (DIM). Cows were blocked by parity and were randomly allocated to either Ovsynch or Heatsynch groups. All cows received 100 microg of GnRH i.m. 14 days after the second set-up injection of PGF(2alpha), followed by a third injection of 25mg PGF(2alpha) i.m., 7 days later. In the Ovsynch group, HAI cows (n=29) were bred on standing estrus after the third PGF(2alpha) before the scheduled second GnRH, whereas FTAI cows (n=218) that were not observed in estrus, received a second injection of 100 microg of GnRH i.m., 48 h after the third PGF(2alpha) and received TAI 8 h after the second GnRH. In the Heatsynch group, all cows (n=288) received 0.5 mg of estradiol cypionate (ECP) 24 h after third PGF(2alpha) and HAI cows (n=172) were bred on standing estrus and FTAI cows (n=116) that were not observed in estrus, received TAI 72 h after the third PGF(2alpha). In Experiment 2, repeat breeder cows (n=186) were randomly assigned to either Ovsynch or Heatsynch groups. The FTAI and HAI cows were inseminated similar to Experiment 1. All cows were observed for estrus three times daily. The associations with the conception rate were modeled with logistic regression separately for Experiments 1 and 2. Of all the variables included in the model in Experiment 1, type of AI (HAI versus FTAI, P=0.0003) and parity (primiparous versus multiparous, P=0.05) influenced the first service conception rate. Over-all conception rate and first service conception rate for HAI cows were higher compared to FTAI cows (33.8% versus 21.3%, and 35.3% versus 21.0%; P=0.001). In the Heatsynch group, cows that received HAI had significantly higher over-all conception rate and first service conception rate compared to FTAI (35.2% versus 17.3% and 36.0% versus 15.5%; P=0.0001). The conception rates in repeat breeder cows for HAI and FTAI (30.1% versus 22.3%) were not different (P>0.1). In conclusion, it was recommended to include AI at observed estrus and fixed-time AI for cows not observed in estrus in order to improve the conception rate in synchronization protocols.     

Pregnancy rate following GnRH + PGF 2alpha treatment of low body condition, anestrous Bos taurus by Bos indicus crossbred cows during the summer months in a tropical environment

Anestrous and lactating Bos taurus by Bos indicus crossbred cows with minimum body condition were studied to determine the efficacy of GnRH+PGF 2alpha combinations for induction of estrus and/or ovulation on pregnancy rate during the months of the year when temperatures are greater. On day 0 (start of treatment), cows were assigned randomly to either treatment or control groups. Treated cows (n = 74) received i.m. 200 microg of GnRH on day 0 and 150 microg of PGF 2alpha 7 days later (day 7). On day 7, treated cows were equally distributed to each of three protocols: (1) Select Synch (n = 25), artificial insemination (AI) 12 h after exhibiting estrus from day 7 (PGF 2alpha injection) until day 12; (2) Ovsynch (n = 24), 200 microg of GnRH at 48 h after PGF 2alpha (day 9) + timed-AI (TAI) 16-20 h later; (3) CO-Synch (n = 25), 200 microg of GnRH + TAI at 48 h after PGF 2alpha (day 9). Control cows (n = 25) received no treatment + AI 12 h after exhibiting estrus from days 0 to 12. Detection of estrus was performed daily during the early morning and evening hours from days 0 to 7 in all the cows, and from days 7 to 12 in the cows treated with Select Synch and in the control group, with the aid of a sterilized bull. Palpation per rectum and transrectal ultrasonography were used on days -30, -20, -10 and 0 to confirm anestrus (absence of CL and no signs of estrus at each evaluation) but with ovarian follicles > or = 10 mm on day 0. Pregnancy rate was 0% for Select Synch, 21% for Ovsynch and 28% for CO-Synch (P < 0.05). In conclusion, the Ovsynch and CO-Synch protocols resulted in greater pregnancy rates compared with the Select Synch protocol in Bos taurus/Bos indicus cows with minimum body condition that were anestrous and lactating during the summer months in a tropical environment.     

A progesterone-based timed AI protocol more effectively prevents premature estrus and incomplete luteal regression than an Ovsynch protocol in lactating Holstein cows

The objective of this study was to evaluate pregnancy rates in lactating Holstein cows treated with an Ovsynch protocol (GnRH-PGF(2alpha)-GnRH) or a progesterone-based timed AI (TAI) protocol, and to determine the factors that may influence pregnancy rate following protocol treatment. In experiment 1, lactating Holstein cows were randomly assigned to three treatments: (1) an injection of GnRH (Day 0), an injection of PGF(2alpha) on Day 7, a second injection of GnRH on Day 9, and TAI 16h after the second GnRH injection (GPG group, n = 34); (2) insertion of a CIDR intravaginal progesterone (1.9g) device combined with a capsule containing 10mg estradiol benzoate (Day 0), an injection of PGF(2alpha) and removal of the device on Day 7, an injection of GnRH on Day 9, and TAI 16h after the GnRH injection (CPG group, n = 34); (3) an injection of PGF(2alpha) after confirming the presence of CL by ultrasonographical observation and artificial insemination at estrus (AIE) (P group, n = 75). The pregnancy rate after TAI following the CPG protocol (41.2%) was higher (P<0.05) than that after TAI following the GPG protocol (20.6%) and that after AIE (20.0%). In experiment 2, lactating Holstein cows were randomly assigned to two treatments: a GPG group (n = 31) and a CPG group (n = 31). The GPG and CPG protocols were identical to those used in experiment 1. The proportion of cows with premature estrus prior to injection of PGF(2alpha) and with incomplete luteal regression tended (P = 0.056) to be or were greater (P<0.05) in the GPG group (4/31, 8/31) than in the CPG group (0/31, 2/31), respectively. Average diameters of dominant follicles (1.5+/-0.1mm versus 1.4+/-0.1mm) on Day 7 and preovulatory follicles (1.8+/-0.1mm versus 1.6+/-0.1mm) on Day 9, and the proportion of cows with synchronized ovulation by 40h after the second GnRH injection were not different (81.5% versus 87.1%, P>0.05) between groups, respectively. We conclude that the pregnancy rate after TAI following the CPG protocol was higher than that after TAI following the GPG protocol, probably due to a decreased incidence of premature estrus and incomplete luteal regression.     

Effects of pre-synchronization using combinations PGF(2alpha) and (or) GnRH on pregnancy rates of Ovsynch- and Cosynch-treated lactating Holstein cows

In Experiment 1, the effects of two pre-synchronization treatments on synchronized AI pregnancy rates of lactating dairy cattle were compared. Lactating Holstein cows (n=159) received 100 microg of GnRH (im) on day -7 and 25mg of PGF(2alpha) (im) on day 0 and were observed once daily for signs of estrus from day -3 to day 3. Cows detected in standing estrus and those that had lost significant amounts of tail-chalk in the previous 24h were immediately inseminated in a once-daily observation/AI program. Cows not detected in estrus by 72 h after PGF(2alpha) received fixed-time AI (TAI) and a concurrent 100 microg injection of GnRH (im). Cows were randomly assigned by parity and calving date to receive one of the following pre-synchronization treatments: (1) 25mg of PGF(2alpha) (im) on day -35 and day -21 (PGF-PGF) or (2) 100 microg of GnRH (im) on day -14 (GnRH). Fewer (P<0.05) GnRH- (49%, 41/84) than PGF-PGF-pretreated cows (65%, 49/75) were detected in estrus, however, overall pregnancy rates were not affected by pre-synchronization treatment (30 versus 32%, respectively). In Experiment 2, lactating Holstein cows received 100 microg of GnRH (im) on day -7, 25mg of PGF(2alpha) (im) on day 0 and TAI at 60-64 h after PGF(2alpha). Cows were randomized by parity and postpartum interval into pre- and post-synchronization treatments in a 2 x 2 factorial design. Pre-synchronization treatments included: (1) 25mg of PGF(2alpha) (im) on day -35 and on day -21 (PGF-PGF; n=168) or (2) 25mg of PGF(2alpha) (im) on day -21 and 100 microg of GnRH (im) on day -14 (PGF-GnRH; n=180). Within each pre-synchronization treatment, cows were further allocated by parity and postpartum interval to receive as a post-synchronization treatment 100 microg of GnRH (im) at either 48 h (Ovsynch; n=175) or 60-64 h (Cosynch; n=173) after PGF(2alpha). Pregnancy rates at TAI were not affected by pre- (PGF-PGF=26%, 44/168 versus PGF-GnRH=24%, 44/180) or post-synchronization treatments (Ovsynch=29%, 50/175 versus Cosynch=22%, 38/173). However, the numeric shift towards reduced pregnancy rates in Cosynch-treated cows suggests the 12h interval between GnRH and AI may be important to optimize conception rates in GnRH-PGF(2alpha)-based TAI protocols in dairy cattle. In conclusion, each of the pre-synchronization protocols evaluated in present study performed with comparable efficacy. Although the Cosynch protocol facilitates more efficient labor utilization, numeric trends toward reduced conception warrants further investigation.     

Incidence of premature estrus in lactating dairy cows and conception rates to standing estrus or fixed-time inseminations after synchronization using GnRH and PGF(2alpha).

Fixed-time AI (TAI) after GnRH-PGF(2alpha)-GnRH treatment is a method to achieve pregnancies in dairy herds without estrous detection. However, cows that fail to respond to the initial GnRH may have compromised TAI conception rates due to asynchronous ovarian response. This study documented the percentage of GnRH-treated Holstein cows (n=345) in two herds that displayed estrus at an inopportune time for optimum TAI conception rate (< or =48h post-PGF(2alpha); premature estrus (PE)) and compared conception rates of two TAI protocols in cows that did not display PE. At biweekly herd health exams, cows diagnosed as not pregnant to a previous AI and cows >80 days postpartum with no AI were treated with 100 microg GnRH (day -7) and 25mg PGF(2alpha) (day 0). Cows detected in PE by twice-daily visual observation from day -7 to day 2 were bred by AI 8-12h later. Cows not detected in PE were randomly assigned by parity, body condition score, and postpartum interval to receive either: (1) 100microg GnRH at 48h after PGF(2alpha) and TAI 16 to 18h later (Ovsynch); or (2) TAI at 72h post-PGF(2alpha) and a concurrent 100 microg GnRH injection to those cows not detected in estrus between 48 and 72h post-PGF(2alpha) (modified Ovsynch (MOV)). All hormone injections were im. Twenty percent (68/345) of the cows were detected in estrus before 48 after PGF(2alpha), of which 5% (17/345) were detected in estrus before PGF(2alpha) (< or =day 0). Herd influenced the percentage of cows in the PE group (herd A versus herd B; 25% versus 14%; P<0.05). Conception rates were not affected by treatment (PE versus Ovsynch versus MOV; 32% (21/65) versus 30% (37/125) versus 32% (47/145); P>0.10). However, within MOV-treated cows, conception rates were greater (P<0.05) in cows detected in estrus (46% (23/50)) compared with cows not detected in estrus (25% (24/95)). In conclusion, 20% of GnRH-treated cows displayed PE and necessitates estrous detection during this period if maximal pregnancy rates are to be achieved. Although additional estrous detection is required compared to Ovsynch, reduced cow handling and hormone usage, efficient use of expensive semen through greater conception rates in cows detected in estrus, and comparable TAI conception rates, suggests the MOV protocol may be a cost effective alternative to Ovsynch in many dairy herd reproductive management programs.     

Ultrasonographic evaluation of endometrial thickness near timed AI as a predictor of fertility in high-producing dairy cows

The objectives were to evaluate changes in endometrial thickness (ET) near the time of a synchronized ovulation and to assess the relationship of ET and fertility in lactating Holstein cows, with or without estrogen supplementation near timed ovulation. In Experiment 1, eight cows were examined with transrectal ultrasonography, once daily for 5 d, starting concurrent with PGF_2α (PGF) treatment during an Ovsynch protocol (GnRH - 7d - PGF - 72h - GnRH). The ET increased rapidly after PGF (from ∼7 to ∼9.5 mm), remained > 9 mm for the next 2 d, then decreased to ∼8 and 7.4 mm, 1 and 2 d, respectively, after the second GnRH. In Experiment 2,642 cows (total of 758 breedings) were subjected to an Ovsynch protocol (GnRH - 7d - PGF - 56h - GnRH - 16h - timed AI); cows received either no further treatment (Ovsynch) or 1 mg of estradiol-17β im 8 h before the second GnRH (Ovsynch + E2). For both uterine horns, ET was measured (∼2 cm from the internal uterine body bifurcation) before E2 treatment (48 h after PGF). In cows with ET ≤ 8 mm vs > 8 mm, rates of ovulation were 86.0% (n = 136) vs 98.1% (n = 472; P < 0.01), respectively, and percentage pregnant per AI (P/AI) were 26.7% (n = 146) vs 42.7% (n = 524; P < 0.01). Treatment with E2 increased P/AI in cows with lower ET (Ovsynch + E2 = 37.0% vs Ovsynch = 23.3%; P = 0.07), but did not significantly improve P/AI in cows with ET > 8 mm (Ovsynch + E2 = 43.4% vs Ovsynch = 42.1%). In conclusion, a single ultrasonographic evaluation of ET in Holstein cows 48 h after PGF treatment in an Ovsynch program was a good predictor of ovulation failure and pregnancy success. Perhaps poor fertility in cows with reduced ET was low peripheral E2 concentrations near AI, poor P4 priming, or luteolysis failure during timed AI procedures.     

Temporal relationships among estrus, body temperature, milk yield, progesterone and luteinizing hormone levels, and ovulation in dairy cows

Estrous cycles of 10 postpartum cyclic Holstein cows were synchronized using prostaglandin f(2alpha) (PGF(2alpha)) given twice 12 d apart to study the relationship of the onset of estrus, body temperature, milk yield, luteinizing hormone (LH) and progesterone concentration to ovulation. Blood samples and body temperatures (vaginal and rectal) were taken every 4 h until ovulation, starting 4 h prior to the second PGF(2alpha) treatment. All cows were observed for estrus following the second administration of PGF(2alpha). Ultrasound scanning of the ovaries commenced at standing estrus and thereafter every 2 h until the disappearance of the fluid filled preovulatory follicle (ovulation). Two cows failed to ovulate and became cystic following the second PGF(2alpha) treatment. The remaining eight cows exhibited a decline in progesterone to <1.0 ng/ml within 28 h, standing estrus and a measurable rise (> 1.0 degrees C) in vaginal but not rectal temperature, and ovulated 90 +/- 10 h after the second PGF(2alpha) treatment. Onset of standing estrus, LH peak and vaginal temperature were highly correlated (P<0.05) with time of ovulation (0.82, 0.81 and 0.74, respectively). Intervals to ovulation tended to depend upon parity. Pluriparous (n = 4) and biparous (n = 4) cows ovulated within 24 and 30 +/- 3 h from the onset of standing estrus; 22 and 31 +/- 2 h from the LH peak; and 22 and 27 +/- 3 h from peak vaginal temperature (mean +/- standard error of the mean), respectively. The results indicated that the onset of standing estrus and rise in vaginal temperature are good practical parameters for predicting ovulation time in dairy cattle.     

Comparison of pregnancy rates with two estrus synchronization protocols in Italian Mediterranean Buffalo cows

The aim in this study was to compare two estrus synchronization protocols in buffaloes. Animals were divided into two groups: Group A (n=111) received 100 microg GnRH on Day 0, 375 microg PGF(2alpha) on Day 7 and 100 microg GnRH on Day 9 (Ovsynch); Group B (n=117) received an intravaginal drug release device (PRID) containing 1.55 g progesterone and a capsule with 10mg estradiol benzoate for 10 days and were treated with a luteolytic dose of PGF(2alpha) and 1000 IU PMSG at the time of PRID withdrawal. Animals were inseminated twice 18 and 42 h after the second injection of GnRH (Group A) and 60 and 84 h after PGF(2alpha) and PMSG injections (Group B). Progesterone (P(4)) concentrations in milk samples collected 12 and 2 days before treatments were used to determine cyclic and non-cyclic buffaloes, and milk P(4) concentrations 10 days after Artificial insemination (AI) were used as an index of a functional corpus luteum. Cows were palpated per rectum at 40 and 90 days after AI to determine pregnancies. All previously non-cyclic animals in Group B had elevated P(4) (>120 pg/ml milk whey) on Day 10 after AI. Accordingly, a greater (P<0.01) relative percentage of animals with elevated P(4) 10 days after AI were observed in Group B (93.2%) than in Group A (81.1%). However, there was no difference in overall pregnancy rates between the two estrus synchronization protocols (Group A, 36.0%; Group B 28.2%). When only animals with elevated P(4) on Day 10 after AI were considered, pregnancy rate was higher (P<0.05) for animals in Group A (44.4%) than Group B (30.3%). The findings indicated that treatment with PRID can induce ovulation in non-cyclic buffalo cows. However, synchronization of estrus with Ovsynch resulted in a higher pregnancy rate compared with synchronization with PRID, particularly in cyclic buffalo.     

Estrus synchronization and artificial insemination of hair sheep ewes in the tropics

Hair sheep ewes (St. Croix White and Barbados Blackbelly) were used to evaluate 3 methods of estrus synchronization for use with transcervical artificial insemination (TAI). To synchronize estrus, ewes (n = 18) were treated with PGF2alpha (15 mg, im) 10 d apart, with controlled internal drug release (CIDR) devices containing 300 mg progesterone for 12 d (n = 18), or with intravaginal sponges containing 500 mg progesterone for 12 d (n = 18). On the day of the second PGF2alpha injection or at CIDR or sponge removal, sterile rams were placed with the ewes. Jugular blood samples were collected from the ewes at 6-h intervals until the time of ovulation, and daily for 16 d after estrus (Day 0). Plasma was harvested and stored at -20 degrees C until LH, and progesterone concentrations were determined by RIA. There was no difference (P>0.10) in time to estrus among the CIDR-, PGF2alpha- or sponge-treated ewes. All of the ewes in the CIDR group and 94.4% of the sponge treated ewes exhibited estrus by 36 h after ram introduction, while only 72.2% of PGF2alpha-treated ewes showed signs of estrus by this time (P<0.06). The time from ram introduction to ovulation was not different (P>0.10) among the CIDR-, PGF2alpha- or sponge-treated ewes. The time to the preovulatory LH surge was similar (P>0.10) among CIDR, PGF2alpha and sponge treated ewes. Progesterone levels through Day 16 after the synchronized estrus were not different (P>0.10) among treatment groups. Hair sheep ewes (n = 23) were synchronized using PGF2alpha and bred by TAI using frozen-thawed semen 48 h after the second injection. The conception rate to TAI was 2/23 (8.7%) and produced 3 ram lambs. In a subsequent trial, 17 ewes were synchronized with CIDR devices and bred by TAI using frozen-thawed semen 48 h after CIDR removal, resulting in a conception rate of 52.9% (9/17). It is possible to synchronize estrus in hair sheep using either CIDRs, sponges or PGF2alpha. Even though there were no significant differences in the timing of ovulation or the LH surge among the treatment groups, a higher conception rate was achieved in ewes synchronized with CIDR devices during the second trial. This may reflect an increase in the skill level of the TAI technician.