Effects of cloprostenol, human chorionic gonadotropin and estradiol benzoate treatment on estrus synchronization in dairy cows

Two consecutive experiments were conducted. In Experiment 1, 24 Friesian lactating cows were randomly assigned to two groups. Cows in Group I received intramuscularly (i.m.) 500 mcg of cloprostenol, 1250 IU of human chorionic gonadotropin (hCG) and 5 mg of estradiol benzoate 12 h after cloprostenol treatment. Cows in Group II received 750 IU i.m. of hCG and 3 mg of estradiol benzoate 12 h after cloprostenol treatment. Treatment was given on Day 16 after estrus in both groups. All animals showed estrus within 24 to 48 h after cloprostenol treatment. The average interval from cloprostenol injection to the onset of estrus was not influenced by treatments. Four cows in Group I failed to ovulate and became cystic. In Experiment 2, 71 Friesian lactating cows were randomly assigned to two groups. Cows in Group I received 500 mcg i.m. of cloprostenol after corpus luteum detection by palpation per rectum. Cows in Group II received 500 mcg of cloprostenol plus 750 IU of hCG and 3 mg of estradiol benzoate 12 h after. When estrus ready for service was confirmed by rectal examination, cows were inseminated. The percentage of cows ready for service tended to be lower (P < 0.06) between cows in Group I (88%) and those in Group II (100%). The average interval from cloprostenol treatment to service was longest (P < 0.001) in Group I (78.7 h +/- 14.9, X +/- SD) vs Group II (48 h +/- 2.9). The degree of readiness for service synchrony was lowest (P < 0.001) in Group I (59.3%) vs Group II (94.2%). The pregnancy rates of cows synchronized or treated were not altered by hCG-estradiol benzoate treatment (P > 0.25). These results suggest that in dairy cows treated with cloprostenol following palpation per rectum of a corpus luteum and then with 750 IU of hCG and 3 mg of estradiol benzoate 12 h later, a single fixed-time insemination at 48 h after cloprostenol treatment should be performed.     

Short synchronization system for estrus cycles in dairy heifers: a preliminary report

In previous studies we demonstrated that the administration of a luteolytic dose of cloprostenol to dairy cows in luteal phase, followed by hCG plus estradiol benzoate (EB) 12 h later, led to successful timed AI 48 h after the initiation of treatment. This article reports two consecutive studies. In Study 1 we determined the pregnancy rate of dairy heifers in luteal phase (established by palpation per rectum) treated with cloprostenol followed by 250 IU of hCG plus 1 mg of EB 12 h later, and inseminated 48 h after cloprostenol injection. Study 2 was designed to evaluate the efficiency this synchronization protocol, irrespective of the estrus stage of the animals. In Study 1, 1272 Friesian heifers aged 14 to 16 months with a palpable corpus luteum received 500 mcg cloprostenol. Heifers were then synchronized either according to the hCG plus EB protocol (hCG-EB, n=637), or by a second dose of cloprostenol 11 d later (PG, n=636). Animals in this last group served as controls and were inseminated 72 and 96 h after the second cloprostenol injection. The pregnancy rate was significantly higher (P<0.0001) in the hCG-EB group (59.5%, 379/637) than in PG (44.8%, 285/636). In Study 2, 135 contemporary heifers (with no corresponding information on estrus stage) were subjected to the same protocol as those in the hCG-EB group of Study 1. These animals were classified in retrospect according to estrus stage established by plasma progesterone concentration. Pregnancy rates were 66.7% (24/36), 51% (25/49) and 58% (29/50) for animals in the follicular, early/late luteal, and mid-luteal phase, respectively. The total pregnancy rate was 57.8% (78/135). These findings indicate an improved pregnancy rate for heifers subjected to single insemination after cloprostenol/hCG/EB synchronization, compared to double insemination after synchronization by 2 cloprostenol injections 11 d apart. The cloprostenol/hCG/EB protocol did result in acceptable pregnancy rates after timed AI of dairy heifers regardless of their estrus cycle phase.     

Compared to dinoprost tromethamine, cloprostenol sodium increased rates of estrus detection, conception and pregnancy in lactating dairy cows on a large commercial dairy

Using two PGF_2α treatments 14 days apart as a way to enhance estrus detection rate following the 2nd treatment is a reproductive management tool that continues to be used on large dairy farms. In one study, in cows with a functional CL and a dominant follicle, treatment with cloprostenol vs. dinoprost resulted in greater peripheral estradiol concentrations. The objective of the present study was to determine if cloprostenol could enhance pregnancy rates of cows in a large dairy herd using a PGF_2α program for 1st artificial insemination (AI). Lactating dairy cows (n = 4549) were randomly assigned to receive two treatments of either 500 μg cloprostenol or 25 mg dinoprost 14 days apart, with the 2nd treatment on the 1st day of the voluntary waiting period (57 DIM). Cows detected in estrus within 5 days after the 2nd treatment were inseminated. There was no effect of treatment on day of estrus detection, with 78% of cows inseminated on Days 3 or 4 following treatment. Cloprostenol increased (P < 0.01) estrus detection rates in 1st parity cows compared to dinoprost, 42.4 vs. 34.0%. In cows inseminated on Days 3 or 4 after treatment, cloprostenol increased (P = 0.05) conception rates compared to dinoprost, 38.3 vs. 34.4%. When treatments and parities were combined, conception rates increased (P < 0.02) with interval after treatment (27.0, 36.4, and 44.5% for Days 1 or 2, Days 3 or 4, and Day 5, respectively). Cloprostenol increased (P = 0.02) overall pregnancy rate compared to dinoprost, 14.4 vs. 12.2%. In summary, cloprostenol increased fertility in 1st parity cows inseminated on Days 3 or 4 following treatment and subsequently enhanced pregnancy rates of 1st parity lactating dairy cows compared to dinoprost. Fertility appeared greater in cows expected to have had a young antral ovarian follicle at treatment.     

Reproductive performance of lactating dairy cows and heifers resynchronized for a second insemination with an intravaginal progesterone-releasing device for 7 or 8d with estradiol benzoate injected at the time of device insertion and 24h after removal

One aim of this study was to compare the reproductive performance of cows and heifers when resynchronizing returns to estrus for a second insemination by treating with an intravaginal progesterone-releasing device (IVD) for 7 or 8d when estradiol benzoate (EB) was administered at the start of treatment and again 24h after device removal. An additional aim was to document the pattern of onset and characteristics of estrus with each resynchrony treatment. Lactating cows in three herds were synchronized for a first estrus and AI by treatment with an IVD for 8d, starting on Day 0, cloprostenol (0.5 mg im) at device removal and EB at device insertion (2.0 mg im) and 24h after removal (1.0 mg im). Cows were resynchronized for a second estrus starting on Day 23 by reinsertion of IVDs for 7 (IVD-7-EB; n=449) or 8d (IVD-8-EB; n=445) with EB (1.0 mg im) administered at device insertion and 24h after removal. Cows were resynchronized for a third estrus by administration of EB (1.0 mg im) on Day 46, but subsequent treatments (no further treatment, reinsertion of CIDR or administration of EB on Day 55) varied among herds as part of separate studies. Maiden heifers (7-Day, n=68; 8-Day, n=69) were similarly treated as cows in a separate herd, but doses of EB were always 1.0 mg im at device insertion and 0.75 mg im 24h after removal. Heifers were not resynchronized for a third estrus. Cattle were inseminated on detection of estrus at each synchronized estrus. Cumulative pregnancy rates 4 week (66.0%, 276/418 versus 59.1%, 247/418) and 7 week (72.7%, 304/418 versus 67.7%, 283/418) after the start of AI were greater (P<0.05) in the IVD-7-EB cows compared to the IVD-8-EB cows, respectively; this was associated with a 9% increase in conception rates at the second estrus (P=0.051) in the IVD-7-EB cows. Treatment did not significantly affect reproductive performance in heifers. Characteristics of estrus measured with radiotelemetry did not differ significantly between the two treatment groups, but more cows were detected in estrus 36 h after removal of IVDs in the IVD-8-EB cows compared to the IVD-7-EB cows (P<0.05). We concluded that reproductive performance in resynchronized dairy cows but not heifers was greater following resynchronization of estrous cycles after AI with an IVD for 7 compared to 8d when EB was injected at the start of treatment and 24h after device removal.     

Reproductive performance of lactating dairy cows treated with cloprostenol at the time of insemination

The effect of intravenous cloprostenol treatment at the time of insemination on reproductive performance was consecutively evaluated in three different subpopulations of high producing lactating dairy cows: Study (1) early postpartum synchronized and fixed-time inseminated (about 50 days in milk) cows (n = 379: 187 control and 192 treated cows); Study (2) presumed high fertility cows first inseminated between 90 and 120 days postpartum (n = 248: 124 control and 124 treated cows); and Study (3) heat stressed repeat breeder cows (n = 183: 93 control and 90 treated cows). Data were analyzed using multiple regression methods. Study 1: Parity (primiparous versus multiparous), milk production, body condition score at AI, insemination season (cool versus warm period) and treatment were included in the analysis as potential factors affecting ovulation, double ovulation, return to estrus, and pregnancy to first AI and to second AI (first AI plus return AI) rates. Logistic regression analysis indicated that the final model for ovulation rate only included the interaction (P = 0.002) between insemination season and treatment. Cloprostenol treatment at insemination led to a 4.2-fold increase in the ovulation rate in cows inseminated during the warm period. There were no significant effects of treatment, parity, milk production, body score or the insemination season on the return to estrus rate. The only variables included in the final logistic model for double ovulation and pregnancy to first AI rates were treatment and season, respectively. Treatment led to a 2.6-fold increase (P = 0.001) in the double ovulation rate, whereas cows inseminated in the warm period were 2.1 times less likely (P = 0.007) to become pregnant at first AI compared to those inseminated in the cool season. The variables included in the final logistic model for the pregnancy rate to second AI were treatment and season. Cloprostenol given at AI increased the risk of pregnancy 1.9 times (P = 0.002), and cows inseminated during the warm season were two times less likely to become pregnant (P = 0.003). No significant interactions were found among these three dependent variables (double ovulation and pregnancy to first and to second AI rates). Study 2: Logistic regression analysis of all the dependent variables: return to estrus, and pregnancy to first and to second AI (first AI plus return to AI) rates indicated no significant effects of treatment, parity, days in milk, milk production or body score at AI. No significant interactions were found. Study 3: The final model for the pregnancy rate only included the interaction between parity (primiparous versus multiparous) and treatment. Days in milk, milk production and insemination number showed no significant effect on pregnancy rate. Cloprostenol treatment at insemination increased the pregnancy rate in primiparous repeat breeder cows (odds ratio: 3.6). The treatment group and parity showed significant (P < 0.0001) interaction. This interaction suggests that cloprostenol treatment of primiparous cows at insemination might enhance pregnancy yet have no effect in multiparous cows. Our findings indicate that cloprostenol administered at insemination promotes ovulation and double ovulation in lactating dairy cows. Cloprostenol treatment showed no benefit in cows with acceptable reproductive performance, suggesting that cloprostenol treatment at AI may only be useful in cows in which stress factors affect ovulation and in repeat breeder cows.     

Comparison of synchronization of ovulation and induction of estrus as therapeutic strategies for bovine ovarian cysts in the dairy cow

The benefit of using timed-insemination in lactating dairy cows for the treatment of ovarian cysts lies in the fact that cows do not have to be detected in estrus for insemination and achieving pregnancy. We compared the effectiveness of synchronization of ovulation with timed-insemination and induction of estrus with insemination at estrus in the treatment of bovine ovarian cysts in lactating dairy cows. After Day 65 post partum, a total of 368 lactating dairy cows was divided into 3 groups. Cows in Group 1 (n = 209, normal, noncystic) were treated with 100 ug, i.m. GnRH on Day 0; 25 mg, i.m. PGF2 alpha on Day 7; and 100 ug, i.m. GnRH on Day 9 and then were time-inseminated 16 h later. Cows in Group 2 (n = 76, abnormal, cystic) were treated with 100 ug, i.m. GnRH on Day 0; 25 mg, i.m. PGF2 alpha on Day 7; and 100 ug, i.m. GnRH on Day 9 and time-inseminated 16 h later. Cows in Group 3 (n = 83, abnormal, cystic) were treated with 100 ug, i.m. GnRH on Day 0; 25 mg, i.m. PGF2 alpha on Day 7; and inseminated at induced estrus within 7 d after treatment with PGF2 alpha. Day 0 was the day of initiation of the study. Conception and pregnancy rates among groups were compared using logistic regression and adjusted for parity, time of year and days in milk. Conception and pregnancy rates of Group 1 cows (31.5%) were not significantly different from those of Group 2 cows (23.6%). However, the pregnancy rate in normal cows (Group 1) was higher (P < 0.01) than in cystic cows (Groups 2 and 3). Cows in Group 3 had a higher conception rate than cows in Group 2 (51.7% > 23.6%; P < 0.01). However, pregnancy rates for cows in Groups 2 (23.6%) and 3 (18%) were not significantly different. The finding indicated that synchronization of ovulation and timed-insemination resulted in pregnancy rates similar to those of synchronization of estrus and insemination at an induced estrus within 7 d for the treatment of ovarian cysts in lactating dairy cows.     

Synchronization of estrus with PGF2 alpha administered 18 days after a progesterone treatment in lactating dairy cows

In a previous study we showed that estrus synchronization with 2 treatments of PGF2 alpha 13 d apart reduced conception rate at the synchronized estrus and that this reduction occurred mainly in cows in the early luteal phase at the second PGF2 alpha treatment. The objective of the present study was to determine the efficacy of a synchronization regimen in which PGF2 alpha was administered during the mid- to late-luteal phase to cows that had previously been synchronized with progesterone. Spring-calving cows from 6 dairy herds were used in this study. On Day -32 (Day 1 = the start of the breeding season), cows that had calved 2 or more weeks ago were randomly assigned to a synchronization (S, n = 732) or control (C, n = 731) group. Cows in Group S were treated with an intravaginal progesterone device (CIDR) for 12 d from Day -32 to Day -20, while those in Group C were left untreated. Similar percentages of cows in Group S (80.6%) and C (82.9%) had cycled by Day -7. The CIDR treatment synchronized the onset of estrus, resulting in 92.9% of cows in estrus being detected within 7 d after CIDR removal. Cows in Group S that had cycled by Day -7 were treated with PGF2 alpha (25 mg, i.m., Lutalyse) on Day -2. Cows in both groups that were anestrous on Day -7 were treated with a combination of progesterone and estradiol benzoate (EB) to induce estrus and ovulation (CIDR and a 10 mg EB capsule on Day -7, CIDR removal on Day -2, and injection of 1 mg EB 48 h after CIDR removal). The PGF2 alpha treatment synchronized the onset of estrus in 87.5% of the cows. Group S and C cows had similar conception rates to first (61.0 vs 58.3%) and second (58.4 vs 60.9%) AI; similar pregnancy rates over the AI period (82.8 vs 79.2%) and over the whole breeding season (91.9 vs 90.6%); and required a similar number of services per pregnancy to AI (1.7 vs 1.8). The interval from the start of the breeding season to conception for cows conceiving to AI or to combined AI and natural mating was shorter (P < 0.001) by 5.7 and 6.2 d, respectively, for the Group S cows. It is concluded that the treatment regimen tested in the present study achieved satisfactory estrus synchronization, had no detrimental effect on fertility at the synchronized estrus, and shortened the interval from start of the breeding season to conception.     

Effect of sequential treatment with prostaglandin F2 alpha and/or oxytocin on estrus and pregnancy rate of lactating dairy cows

A total of 335 lactating dairy cows was used to determine the effect of oxytocin or PGF2a given 8 h after treatment with a luteolytic dosage of PGF2a on the percentage of cows exhibiting estrus within 7 d after treatment, and the pregnancy rate to a single insemination at this time. On the initial day of treatment (Day 0), cows with a palpable corpus luteum on the ovary were treated with 25 mg, im of PGF2a. At 8 h later, the cows were divided into 3 groups. Cows in Group 1 (n = 112) were treated with oxytocin (0.33 IU/kg bwt im); cows in Group 2 (n = 112) were treated with 25 mg, im of PGF2a; and cows in Group 3 (n = 111) served as the untreated controls. Cows in all 3 groups were continuously observed for estrus visually or by way of an activated heatmount detector within 7 d after treatment, and were inseminated within 12 h of the observed estrus.Plasma progesterone (P4) concentration was determined using radioimmunoassay on Day 0 and Day 2. Of the cows with P4 greater than 1 ng/ml on Day 0, the percentage of cows observed in estrus within 7 d after treatment was 75, 89 and 72% for cows in Group 1, Group 2 and Group 3, respectively. When all cows were evaluated, the percentage of cows observed in estrus within 7 d after treatment was 60, 70 and 55% for cows in Group 1, Group 2 and Group 3, respectively. In both instances, the value for cows in Group 2 was significantly higher than that for either cows in Group 1 or Group 3. The pregnancy rate for cows inseminated within 7 d was similar for cows in all 3 groups. The results of this study demonstrated that treatment of dairy cows with 2 luteolytic dosages of PGF2a at an 8-h interval resulted in more cows being observed in estrus within 7 d than with 1 treatment with PGF2a, or with oxytocin given at an 8-h interval after a luteolytic dosage of PGF2a.     

Reproductive performance of lactating dairy cows treated with gonadotrophin-releasing hormone (GnRH) and/or prostaglandin F2a (PGF2a) for synchronization of estrus and ovulation

The objective of this study was to determine the reproductive performance of lactating dairy cows treated with GnRH and/or PGF2a for synchronization of estrus and ovulation. Between Days 43 and 57 post partum, a total of 374 dairy cows was divided into 4 groups. Cows in Group 1 (n = 62) were treated with 25 mg, i.m. PGF2a on Days 43 and 57; cows in Group 2 (n = 65) were not treated at this time; cows in Group 3 (n = 118) were treated with 100 ug, i.m. GnRH on Day 50, 25 mg, i.m. PGF2a on Day 57, 100 ug, i.m. GnRH on Day 59, and time-inseminated 16 h later; cows in Group 4 (n = 129) were treated with 25 mg, i.m. PGF2a once on Day 57. Cows in Groups 1 and 4 were inseminated at an induced estrus within 7 d after the last PGF2a treatment, and cows in Group 2 were inseminated at a noninduced estrus within a corresponding period of time. Conception rate, estrus detection rate and pregnancy rate were analyzed using logistic regression, and controlled for lactation number, body condition score and time of year. Days from calving to conception were analyzed using the GLM procedures of SAS, and the model included group, body condition score, lactation number, time of year, and their interactions. Cows in Group 3 had a significantly higher pregnancy rate than cows in Groups 1, 2 and 4. Orthogonal contrasts of mean days from calving to conception showed that cows in Group 3 had significantly (P < 0.01) less days from calving to conception than cows in Group 1 and Group 4. There was a significant effect of time of year on pregnancy rate and days from calving to conception, but there was no interaction between time of year and these reproductive characteristics. There was no effect of body condition score and lactation number on the reproductive characteristics evaluated. From the results of this study, it was concluded that better reproductive performance was observed in cows inseminated at a synchronized ovulation than in those inseminated at a synchronized estrous period.     

Comparison of two estrus synchronization and resynchronization treatments in lactating dairy cows

Reproductive performance in cows following synchronization of estrus with intravaginal progesterone releasing devices (IVD) has varied with the length of treatment, cyclic status and prolonged return to estrus intervals in some cows following first AI. The objective of this study was to compare two methods of synchronizing and resynchronizing estrus on the reproductive performance of lactating dairy cows. Cows were treated with an IVD (Day 0) for 7 days (n = 350) or 8 days (n = 350), cloprostenol (0.5 mg i.m.) at the time of device removal and estradiol benzoate (EB) at the time of device insertion (1.5mg i.m.), and again 9 days later (1.0 mg i.m.). Cows were also resynchronized starting on Days 23 and 46 by reinsertion of IVDs for either 7 or 8 days and treatment with EB (1mg i.m.) at the time of device insertion and again 9 days later. Cows were inseminated on detection of estrus for 4 days after removal of devices at each of the synchronized estrous cycles. No significant differences in reproductive performance were detected between each treatment throughout the study period. Synchrony of estrus was more precise at the first and second estrus after treatment with an IVD for 8 days compared to 7 days. Cows classified as anestrous had lower reproductive performance than cows classified as cycling and had longer intervals to estrus at the second (P < 0.001) and third estrus (P < 0.06), but not at the first estrus (P = 0.09). Mean time to onset of estrus after IVD removal was less in cows treated with an IVD for 8 days compared to 7 days at each synchronized estrus (P < 0.01). More Holstein-Friesian cows were classified as non-pregnant and not detected in estrus than crossbreed cows (15.7%, 54/343 versus 9.0%, 24/266; [P < 0.05). The results of the study suggested that the main effects of the treatments that were used to synchronize and resynchronize estrus were to alter the timing and synchrony of estrus without affecting fertility.     

Reproductive performance of dairy cows with ovarian cysts after different GnRH and cloprostenol treatments

Cystic ovarian disease is an important cause of reproductive failure and economic loss for the dairy industry. This report describes two consecutive studies. The objective of the first was to evaluate the response of cows with ovarian cysts to two therapeutic treatments. In the second study, we compared the effectiveness of the best treatment established in Study 1 with that of the Ovsynch protocol. For Study 1, cows were considered to have an ovarian cyst if it was possible to observe a single follicular structure with a follicular antrum diameter > 25 min in the absence of a corpus luteum in three ultrasonographic examinations performed at 7 days intervals. At diagnosis (Day 0), cows were assigned to one of two treatment groups. Cows in Group GnRH/CLP (n = 31) were treated with 100 microg GnRH i.m. and 500 microg cloprostenol (CLP) i.m. on Day 14. Cows in Group GnRH-CLP/CLP(n = 32) were treated with 100 microg GnRH i.m. plus 500 microg CLP i.m. on Day 0, and 500 microg CLP i.m. on Day 14. The animals were inseminated at observed estrus and monitored weekly by ultrasonography for 4 weeks or until Al. Cows in the GnRH-CLP/CLP group showed a lower cystic persistence rate (15.6% < 45.2%; P = 0.01); a higher estrus detection rate (84.4% > 41.9%; P < 0.0001); a higher ovulation rate (75% versus 32.3%; P < 0.0001) and a higher early response rate (31% > 3%; P = 0.02) than those in the GnRH/CLP group. For the second study, 128 cows with ovarian cysts were randomly assigned to one of two treatment groups: cows in Group Ovsynch (n = 64) were treated with 100 microg GnRH i.m. on Day 0, 500 microg CLP on Day 7, and 100 microm GnRH i.m. 36 h later. Cows in this group were inseminated 24 h after the second GnRH dose (Ovsynch protocol). Cows in Group GnRH-CLP/CLP/GnRH (n = 64)were treated as those in the GnRH-CLP/CLP group of Study 1 but received GnRH 32 h after the second CLP treatment and were inseminated 24 h after this. A further group of cows without ovarian cysts inseminated at natural estrus served as the Group Control (n = 64). Cows in the GnRH-CLP/CLP/ GnRH group showed a lower cystic persistence rate (10.9% < 46.9%; P < 0.0001); higher ovulation rate (79.7% > 17.2%; P < 0.0001); higher return to estrus rate (34.3% > 12.5%; P < 0.01) and higher pregnancy rate (28.1% > 3.1%; P < 0.01) than those in Ovsynch; and a similar pregnancy rate (28.1% versus 35.9%) to Control cows. These findings indicate that lactating cows with ovarian cysts can be successfully synchronized and time inseminated using a protocol that combines GnRH and CLP, starting treatment by simultaneously administering both products. This protocol also allows the insemination of cows showing estrus within the first week of treatment. Ovarian cysts were less responsive when treatment was started with GnRH alone.     

Comparison of pregnancy rates of repeat-breeder dairy cows given gonadotropin releasing hormone at or prior to the time of insemination

A total of 585 repeat-breeder dairy cows was used to study the effect of GnRH treatment, either at or prior to insemination, on the pregnancy rate. The cows were divided into 6 treatment groups. Cows in Group 1 (n = 142) were observed in estrus, and 11 +/- 0.42 hours (mean +/- SEM) later they were given 100 ug, i.m. gonadotropin releasing hormone (GnRH) and were inseminated. Cows in Group 2 (n = 139) were observed in estrus and were inseminated 11.4 +/- 0.43 hours later. Cows in Group 3 (n = 33) were monitored for estrus with an activated heatmount detector but were not observed in estrus; they were inseminated 1.5 +/- 0.87 hours later and were given 100 ug, i.m. GnRH. Cows in Group 4 (n = 35) were not observed in estrus, but they did activate the heatmount detector and were inseminated 2.2 +/- 0.87 hours later. Cows in Group 5 (n = 107) were observed in estrus, given 100 ug, i.m. GnRH 2.0 +/- 0.40 hours later, and were inseminated 9 +/- 0.60 hours after GnRH treatment. Cows in Group 6 (n = 129) were observed in estrus and were inseminated 10 +/- 0.50 hours later. Pregnancy rates were analyzed by Chi-square. Interactions between pregnancy rate, treatment and time of insemination were evaluated using ANOVA and LSM (P < 0.05). There was no effect on pregnancy rate when GnRH was given at or prior to insemination. Cows inseminated on the basis of observed estrus had a higher pregnancy rate (P < 0.05) than cows inseminated on the observation of an activated heatmount detector. From the results of this study, it is concluded that treatment with GnRH at or prior to insemination did not improve the pregnancy rate of repeat-breeder dairy cows.     

Rates of luteolysis and pregnancy in dairy cows after treatment with cloprostenol or dinoprost

Our objective was to determine whether rates of luteolysis or pregnancy differed in lactating dairy cows of known progesterone status and either known or unknown luteal status after either cloprostenol or dinoprost was injected as part of a timed-insemination program. In Experiment 1, 2358 lactating dairy cows in six herds were given two injections of PGF_2α 14 d apart (Presynch), with the second injection given 12 to 14 d before the onset of a timed AI protocol (Ovsynch). Cows (n = 1094) were inseminated when detected in estrus after the Presynch PGF_2α injections. Cows not inseminated (n = 1264) were enrolled in the Ovsynch protocol and assigned randomly to be treated with either cloprostenol or dinoprost as part of the timed-AI protocol. In cows having pretreatment concentrations of progesterone ≥ 1 ng/mL and potentially having a functional corpus luteum (CL) responsive to cloprostenol (n = 558) or dinoprost (n = 519), dinoprost increased (P < 0.05) luteal regression from 86.6 to 91.3%. Despite a significant increase in luteolysis, pregnancies per AI did not differ between luteolytic agents (dinoprost = 37.8% and cloprostenol = 36.7%). Fertility was improved in cows of both treatments having reduced concentrations of progesterone at 72 h and in cows showing signs of estrus. In Experiment 2, an ovulation-resynchronization program was initiated with GnRH or saline in 427 previously inseminated lactating dairy cows of unknown pregnancy status in one herd. Seven days later, pregnancy was diagnosed and nonpregnant cows were blocked by number of CL and assigned randomly to be treated with cloprostenol or dinoprost. Compared with cloprostenol, dinoprost increased (P < 0.05) luteal regression from 69.1 to 78.5%, regardless of the number of CL present or the total luteal volume per cow. Pregnancies per AI did not differ between dinoprost (32.8%) and cloprostenol (31.3%). Although dinoprost was more effective than cloprostenol at inducing luteolysis in lactating dairy cows exposed to an Ovsynch or ovulation-resynchronization protocol, resulting fertility did not differ between products.     

Persistent ovarian follicles in dairy cows: a therapeutic approach

Anestrus is common during the postpartum period in high-producing dairy cows. In a previous investigation, we were able to diagnose persistent follicles of 8 to 12 mm in anestrous cows. This report describes 2 consecutive studies. The objectives of the first were to 1) assess the association of persistent follicles with anestrus; and 2) evaluate 2 therapeutic treatments. In the second study, we compared the effectiveness of the best treatment established in Study 1 with the Ovsynch protocol. For Study 1, anestrous cows were considered to have a persistent follicle if it was possible to observe a single follicular structure > 8 mm in the absence of a corpus luteum or a cyst in 2 ultrasonographic examinations performed at an interval of 7 d. At diagnosis (Day 0), cows were assigned to 1 of 3 treatment groups. Cows in Group GnRH/PGF (n=17) were treated with 100 microg GnRH i.m., and 25 mg PGF2alpha i.m. on Day 14. Cows in Group PRID (n=18) were fitted with a progesterone releasing intravaginal device (PRID, containing 1.55 g of progesterone) for 9 d and were given 100 microg GnRH i.m. at the time of PRID insertion, and 25 mg PGF2alpha i.m. on Day 7. Cows in Group Control (n=18) received no treatment. The animals were inseminated at observed estrus and were monitored weekly by ultrasonography until AI or 5 weeks from diagnosis. Blood samples were also collected on a weekly basis for progesterone determination. The mean size of persistent follicles on Day 0 was 9.4 +/- 0.04 mm. Progesterone levels were < 0.2 ng/mL during the first 35 d in 16 of 18 Control cows. Cows in the PRID group showed a lower persistent follicle rate (16.7% < 70.6% < 88.9%; P < 0.0001; PRID vs GnRH/PGF vs Control, respectively); a higher estrus detection rate (83.3% > 29.4% > 11.1%; P < 0.0001) and a higher pregnancy rate (27.8% > 5.9% > 0%; P = 0.02). For the second study, 145 cows with persistent follicles were randomly assigned to 1 of 2 treatment groups: cows in Group Ovsynch (n=73) were treated with 100 microg GnRH i.m. on Day 0, 25 mg PGF2alpha i.m. on Day 7, and 100 microm GnRH i.m. 32 h later. Cows in this group were inseminated 16 to 20 h after the second GnRH dose (Ovsynch protocol). Cows in Group PRID (n=72) were treated as those in the PRID group of Study 1, and were inseminated 56 h after PRID removal. Cows in the PRID group showed a higher ovulation rate (84.8% > 8.2%: P < 0.0001); a higher pregnancy rate (34.2% > 4.1%; P < 0.0001) and lower follicular persistence rate (22.2% < 63%; P < 0.0001) than those in Ovsynch. Our results indicate that persistent follicles affect cyclic ovarian function in lactating dairy cows. Cows with persistent follicles can be successfully synchronized and time inseminated using progesterone, GnRH and PGF2alpha but show a limited response to treatment with GnRH plus PGF2alpha.     

Effects of presynchronization during the preservice period on subsequent ovarian activity in lactating dairy cows

Among the strategies aimed at overcoming difficulties in estrus detection in dairy herds, presynchronization with two PGF2alpha treatments 14 days apart before a timed AI protocol has been related to a significant increase in pregnancy rates. The aim of the present study was to evaluate the effects of presynchronization during the preservice period on subsequent ovarian activity in clinically normal lactating dairy cows. A second objective was to evaluate the incidence of reproductive disorders on Day 50 postpartum. Depending on the chronological order of parturition, cows were alternately assigned to a control (n=102) or treatment (n=101) group. Animals in the treatment group were administered two cloprostenol treatments 14 days apart, beginning on Day 22 postpartum. The reproductive tract of each animal was examined ultrasonographically on Days 43 and 50 postpartum to monitor ovarian structures and uterine contents. Blood samples were collected on Day 50 for progesterone determination. Cows were inspected for signs of estrus between Days 50 and 71 postpartum and were then inseminated. Follicular persistence rates were similar in the presynchronized (14.9%) and control (13.7%) groups. Cows in the presynchronized group showed a lower metritis-pyometra rate (0% < 3.9%; P=0.045); a lower ovarian cyst rate (3% < 10.8%; P=0.03); a higher luteal activity rate (progesterone concentrations>/=1ng/ml) on Day 50 postpartum (76.2% > 52.9%; P=0.0005); a higher estrus detection rate (73.3% > 47.1%; P<0.0001); a higher ovulation rate (72% > 44%; P<0.0001) and a higher pregnancy rate (29.7% > 15.7%; P=0.02) than controls. Our results indicate that presynchronization during the preservice period reduces the incidence of ovarian cysts and metritis-pyometra determined on Day 50, and improves ovarian activity from Days 50 to 71 postpartum along with pregnancy rates in clinically normal lactating dairy cows.     

Effect of presynchronization strategy before Ovsynch on fertility at first service in lactating dairy cows

The aim of this study was to evaluate the effect of presynchronization with or without the detection of estrus on first service pregnancy per artificial insemination (P/AI) and on Ovsynch outcome in lactating dairy cows. A total of 511 cows were divided randomly but unevenly into 3 treatment groups at 44 to 50 days in milk (DIM). Ovsynch was started at the same time (69 to 75 DIM) in all three groups. Cows in the Ovsynch group (CON, N = 126) received no presynchronization before Ovsynch, and all cows were bred by timed AI (TAI). Cows in the presynchronization with estrus detection (PED) and the presynchronization with only TAI (PTAI) groups received two doses of prostaglandin F_2α (PGF) 14 days apart, starting at 44 to 50 DIM. Ovsynch was initiated 11 days after the second PGF treatment. Cows in the PED group (N = 267) received AI if estrus was detected after either PGF injection. Cows that were not determined to be in estrus after PGF injection received Ovsynch and TAI. Cows in the PTAI group (N = 118) were not inseminated to estrus, with all cows receiving TAI after Ovsynch. The ovulatory response to the first GnRH injection administered as part of Ovsynch differed (P = 0.002) among treatment groups (83.1% in PTAI, 72.6% in PED, and 62.7% in CON). However, the ovulatory response to the second injection of GnRH during Ovsynch did not differ among treatment groups. Of the 267 PED cows, a total of 132 (49.4%) exhibited estrus and were inseminated. The P/AI at the 31-day pregnancy diagnosis was similar between the cows in the PED group with AI after estrus detection (37.9%; 50/132) and those bred with TAI (34.1%; 46/135). The P/AI in the CON group (46.8%; 59/126) was greater (P < 0.05) than that in the PED group (36.0%; 96/267). In addition, the P/AI in the CON group was greater (P = 0.04) than that in the PED cows receiving TAI (34.1%; 46/135) but less than that in the PED cows bred to estrus (37.9%; 50/132) (P = 0.16). At the 31-day pregnancy diagnosis, the cows in the PTAI group had greater P/AI (55.9%; 66/118) than both those in the PED group (P < 0.01; either estrus or TAI) and those in the CON group (P = 0.08). Thus, presynchronization with PGF (PTAI) increased the ovulatory response to Ovsynch and improved P/AI in dairy cows. Interestingly, the breeding of cows to estrus during presynchronization reduced fertility to the TAI and overall fertility, including cows bred to estrus and TAI. These results indicate that maximal fertility is obtained when all cows receive TAI after the presynchronization protocol.     

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.     

Reproductive performance of postpartum dairy cows under a highly intervenient breeding program involving timed insemination and combinations of GnRH, prostaglandin F2alpha and human chorionic gonadotropin

Lactating Holstein cows (n=288) were grouped as pairs at parturition and randomly assigned to two treatments (control, C vs intervenient treatment, T). The reproductive management of the Group C cows (n=130) consisted of the intramuscular administration of 500 microg PGF2alpha analogue (PG) on Days 28 and 63 postpartum and breeding on the basis of estrus signs with the a.m.-p.m. rule after Day 63. Cows that were not bred by 77 d postpartum received another injection of PG and were bred at estrus or 84 h after PG treatment. Pregnancy diagnoses were perfomed by palpation of the uterus per rectum 42 to 48 d after AI. Cows in the T group (n=139) received intramuscular injections of 100 microg GnRH 14 d and PG 28 d after calving. On Day 56 postpartum, cows were given a second dose of GnRH followed by PG on Day 63 postpartum and a third GnRH injection 48 h after PG (OvSynch). Cows were inseminated at a fixed time (22+/-1 h) after GnRH. Five days after the fixed-time insemination cows were given 1500 IU hCG i.m.. Group C and T cows that returned to service or were diagnosed as non-pregnant continued to receive PG at intervals of 14 d with breeding at estrus or 84 h after the second PGF2alpha dose. A sustained increase in milk progesterone concentration was observed in 59.0% of T cows after GnRH administration on Day 14. A similar rise in milk progesterone concentrations was observed in 53.8% of C cows. The PG on Day 28 induced luteolysis more in Group T cows (53.2%) than in Group C cows (36.9%). The PG on Day 63 reduced milk progesterone concentrations to basal levels in 50.7% of T and 49.2% of Group C animals. The first service pregnancy rates (T, 40.3% vs C, 36.2%) and the overall pregnancy rates (all services, T, 83.5% vs C, 86.9%) were not different between the two groups. The two treatments did not differ in the interval from first service to pregnancy, calving to pregnancy or in calving interval, number of services per pregnancy or culling rates.     

Comparison of timed insemination with insemination at estrus following synchronization of estrus with a MGA-prostaglandin system in beef heifers

Three trials utilizing 231 beef heifers were conducted in 1993 to determine if a timed insemination would result in similar synchronized pregnancy rates as insemination by estrus following synchronization of estrus using the 14-d MGA-prostaglandin system. All heifers were fed 0.5 mg MGA/h/d fof 14 d and given a 25 mg injection of PGF(2)alpha im 17 d after the final day of MGA feeding. Heifers in Group 1 (timed AI treatment) were inseminated at 72 h after the prostaglandin injection independent of whether or not they were observed in estrus. Heifers in Group 2 (AI by estrus) were inseminated 12 to 18 h after the onset of estrus. Since the trial was a significant source of variation for synchronized pregnancy rate, the effect of treatment on pregnancy rate was analyzed for each trial. Synchronized pregnancy rates in Trials 2 and 3 were similar in both treatment groups; 37 vs 35% and 61 vs 58% for the timed AI vs AI by estrus (Groups 1 and 2) in Trials 2 and 3, respectively. In both of these trials the degree of estrous synchrony was high. In Trial 1, the synchronized pregnancy rate in heifers that were time-inseminated was significantly lower than that of heifers that were inseminated by estrus (29 vs 57%). The lower synchronized pregnancy rate of Group 1 (timed AI) heifers in Trial 1 appeared to be due to the low degree of estrous synchrony in this trial. Our results indicate that using timed insemination with the 14-d MGA-prostaglandin system will give similar synchronized pregnancy rates as inseminating by estrus in groups of beef heifers where the degree of synchrony is high. However, in heifers where the degree of estrous synchrony is low, a timed insemination reduces synchronized pregnancy rates.     

Estrus and pregnancy rate of dairy cows given one or two doses of prostaglandin F2 alpha 8 or 24 hours apart

A total of 329 cows was used in 2 experiments to study the effect of PGF2a given 8 or 24 h apart on the number of cows observed in estrus within 7 d and the pregnancy rate to a single insemination at this time. In Experiment 1, 233 cows were divided into 2 groups. Cows in Group 1 (n = 117) were treated twice with 25 mg, im of PGF2a (0 and 8 h) while cows in Group 2 (n = 116) were treated only once (0 h). In Experiment 2, 96 cows were divided into 2 groups. Cows in Group 1 (n = 49) were treated twice with 25 mg, im of PGF2a (0 and 24 h) while cows in Group 2 (n = 47) were treated only once (0 h). In Experiment 1, blood samples were obtained from cows in both groups on Days 0 and 2. However, in Experiment 2, blood samples were obtained from cows in both groups only on Day 0. Plasma progesterone concentration was determined using radioimmunoassay. Cows in both experiments were observed for estrus within 7 d of treatment and were inseminated within 12 h of the observed estrus. In Experiment 1, more cows in Group 1 were observed in estrus within 7 d than in Group 2 (P <0.03). In Experiment 2, there was no significant difference in the number of cows in both groups that were observed in estrus within 7 d. However, the interval from treatment to the first observed estrus for cows not observed in estrus within 7 d was significantly longer in cows treated at 0 and 24 h compared with the cows treated once at 0 h. In both experiments, the pregnancy rate for cows inseminated within 7 d was similar for cows in all groups. From the results of this study, it is concluded that treatment of dairy cows with 2 luteolytic dosages of PGF2a at an 8-h interval resulted in more cows being observed in estrus than at a 24-h interval.