Pregnancy Rates in Angus Cross Beef Cows Bred at Observed Oestrus With or Without Second GnRH Administration in Fixed‐Time Progesterone‐Supplemented Ovsynch and CO‐Synch Protocols

Crossbred cows (n = 1073) from five locations had oestrous cycles synchronized with 100 μg of GnRH IM and insertion of controlled internal drug release device (CIDR) on Day 0 followed by 25 mg of PGF_2α IM and CIDR removal on Day 7. Kamar^® patches were placed on all cows at CIDR removal. Cows were observed three times daily for oestrus after PGF_2α administration. In the Ovsynch‐CIDR group, cows detected in oestrus (n = 193) within 48 h after PGF_2α were inseminated using the AM–PM rule. Among these cows, 80 received and 113 did not receive a second GnRH at 48 h after PGF_2α. Cows (n = 345) not detected in oestrus received a second GnRH at 48 h after PGF_2α on Day 9, and fixed‐time AI 16 h after the GnRH on Day 10. In the CO‐Synch‐CIDR group, cows detected in oestrus (n = 224) within 48 h after PGF_2α were inseminated using the AM–PM rule. Among these cows, 79 received and 145 did not receive a second GnRH at 64 h after PGF_2α. Cows (n = 311) not detected in oestrus received a second GnRH on Day 10 at the time of AI, 64 h after PGF_2α. The AI pregnancy rates were not different between the Ovsynch‐CIDR and CO‐Synch‐CIDR groups (p = 0.48). There were no differences in the AI pregnancy rates for cows inseminated at a fixed time (p = 0.26) or at detected oestrus (p = 0.79) between the treatment groups. Among cows inseminated in oestrus, there were no differences in the AI pregnancy rates between cows that received or did not receive the second GnRH (p = 0.47). In conclusion, acceptable AI pregnancy rates can be achieved with or without inclusion of oestrus detection in the Ovsynch‐CIDR and CO‐Synch‐CIDR protocols. Among cows detected in oestrus, cows that received a second GnRH yielded similar pregnancy rates when compared with cows that did not receive the second GnRH.     

Timing of Ovulation and Conception Rate in Primiparous and Multiparous Cows after Synchronization of Ovulation with GnRH and PGF2α

Conception rates after Ovsynch have been higher in primiparous than in multiparous cows. The objective of this study was to investigate whether this difference might be due to differences in ovulation rate or follicular size. The experiment was conducted with 136 Holstein Frisian cows from a commercial herd in Brandenburg, Germany. All cows were synchronized using Buserelin (GnRH analogue) at day ?10, Tiaprost (PGF_2α analogue) at day ?3 and again GnRH at day ?1. Timed artificial insemination (TAI) was carried out 16–20 h after the second dose of GnRH on day 0. Milk samples for analysis of milk progesterone were obtained on days ?17, ?10, ?3 and at TAI. Progesterone concentrations were used to determine the stage of oestrus cycle at the start of the synchronization protocol and to investigate the presence of functional luteal tissue before treatment with PGF_2α and TAI. All animals were examined by ultrasound at the second treatment with GnRH, at AI, 8 and 24 h after AI. Overall synchronization rate (proportion of cows with an ovulation within 40 h after GnRH) was 86.8% in primiparous and 88.2% in multiparous cows, respectively. Ovulation occurred earlier in primparous than in multiparous cows (p < 0.05) and ovulatory follicles were smaller. Conception rates were numerically higher in primiparous cows but the difference was not significant. Cows that displayed signs of oestrus on day ?1 and received an additional AI on this day were more likely to conceive than cows that only received TAI 16 to 20 h after GnRH2. It is concluded that ovulation occurs earlier in primiparous than in multiparous cows after Ovsynch. However, a significant relationship between these differences and the probability of conception could not be established.     

The Effect of Dose and Type of Cloprostenol on the Luteolytic Response of Dairy Cattle during the Ovsynch Protocol under Different Oestrous Cycle and Physiological Characteristics

The objective of this study was to characterize the effect of dose and type of cloprostenol (CLO) on the luteolytic response of dairy cattle during the Ovsynch protocol under different oestrus cycle and physiological characteristics. Twelve non‐lactating dairy cows and 111 lactating dairy cows were used in three experiments. In Experiment I, cows were synchronized so that they had only a 5.5‐ to 6‐day‐old corpus luteum (CL) at the time of the prostaglandin F_2α (PGF_2α) treatment of Ovsynch. In Experiment II, cows were synchronized so that they had at least a CL of approximately 14 days old at the time of PGF_2α treatment and an accessory CL if they had responded to the first GnRH of Ovsynch. Furthermore, in each experiment, cows received either a standard or a double dose of d‐CLO as the luteolytic treatment. In Experiment III, lactating cows were blocked by parity and assigned to one of three luteolytic treatments during Ovsynch: 500 μg d,l‐CLO, 150 or 300 μg of d‐CLO. In Experiment I, the dose of d‐CLO had an effect (p = 0.08) on the percentage of cows with full luteolysis, but not in Experiment II (p > 0.1). More cows in Experiment II had full luteolysis than did cows of Experiment I (87% vs 58%, respectively; p = 0.007). In Experiment III, 87.1%, 84.4% and 86.2% lactating dairy cows had full luteolysis and 37.8%, 36.8% and 36.1% of cows became pregnant after treatment with 500 μg d,l‐CLO, 150 or 300 μg of d‐CLO, respectively (p > 0.05).     

Ovsynch Plus protocol improves ovarian response in anovular Murrah buffaloes in low‐breeding season

The objective of the study was to evaluate the efficacy of ovarian response and pregnancy rate in anovular buffaloes following Ovsynch and Ovsynch Plus protocols. Buffaloes (n = 55) were divided into two groups: Ovsynch group (n = 26): GnRH (10 μg, GnRH1) on Day 0, PGF₂α (25 mg) on Day 7, GnRH (10 μg, GnRH2) on Day 9; Ovsynch Plus group (n = 29): 500 IU equine chorionic gonadotropin (eCG) 72 hr (day ?3) prior to Ovsynch protocol, followed by fixed timed artificial insemination (FTAI) 6 and 24 hr after GnRH2 injection in bot groups. Transrectal ultrasonography was performed daily, that is, from day 0 and ?3 in Ovsynch and Ovsynch Plus group, respectively for ovarian response and pregnancy diagnosis at day 30 post‐insemination. In Ovsynch Plus group, administration of eCG prior to GnRH1 increased (p < .001) the diameter (mm) of dominant follicle (DF) from 10.15 ± 0.26 to 12.23 ± 0.34 within 72 hr of treatment resulting higher ovulatory response to GnRH1. Ovulation after GnRH1 was higher (p < .01) in Ovsynch Plus group (96.6%) than Ovsynch group (61.5%). However, ovulation rate to GnRH2 was similar (p > .05) between groups (Ovsynch group: 76.9% vs. Ovsynch Plus group: 70.0%). Mean DF diameter (mm) that ovulated to both GnRHs was higher (p < .01) than non‐ovulated counterparts in both groups (Ovsynch group: 10.80 ± 0.27 vs. 8.47 ± 0.53; Ovsynch Plus group: 11.99 ± 0.24 vs. 9.5 ± 0.63). Pregnancy was established in buffaloes which responded to both GnRHs, irrespective of groups, being higher (p = .52) in Ovsynch Plus group (34.5%) than Ovsynch group (23.1%), though non‐significant. In summary, this study showed that eCG inclusion prior to Ovsynch regimen improves ovulatory response in anovular buffaloes during low‐breeding season.     

Evaluation of three hormonal protocols for anovulatory lactating cows under regulations restricting the use of estrogenic compounds

When European Union regulations restricted the use of estrogenic compounds in food‐producing animals, refined hormonal protocols were no longer applicable for anovulatory cows. However, Ovsynch and its adaptations are routinely and uniformly applied to all cows regardless of ovarian function. To evaluate their efficacy on anovulatory cows, 143, 147 and 144 anovulatory cows received Ovsynch, Presynch and G6G protocols, respectively. In comparison, 150 cyclic cows were bred without using a synchronized protocol. Results showed that cows in the Presynch group had luteolysis responding to the last prostaglandin F_2α (PGF_2α) injection greater than the Ovsynch group. The serous progesterone levels at the first gonadotropin‐releasing hormone of Ovsych and the last PGF_2α injection was greater in the G6G group than the other two hormonal treatment groups. Concentrations of Ca²⁺ and total protein in cervical mucus in all three hormone‐treated groups before artificial insemination (AI) were significantly different from the controls. The G6G group obtained a greater pregnancy rate compared with Ovsynch and Presynch, but significantly less than the controls. For open cows in the Ovsynch group, estrus rate within 24 days after the first AI was significantly less than the controls. In conclusion, the G6G treatment resulted to better reproductive performance in anovulatory cows.     

Strategies to optimize reproductive efficiency by regulation of ovarian function

Pregnancy rate to the Ovsynch protocol can be improved if cows are presynchronized (i.e., two PGF(2alpha) injections given 14 days apart and the second injection of PGF(2alpha) given 12 days prior to the first GnRH of the Ovsynch program) so that a greater proportion of cows during the Ovsynch protocol ovulate to the first GnRH injection and have a CL at PGF(2alpha) injection. Pregnancy rates were normal in anestrous cows (39.6%) if they ovulated to both injections of GnRH. Estradiol cypionate (ECP) can be used to replace GnRH to induce ovulation as a modification of the Presync-Ovsynch program (i.e., Presync-Heatsynch). Pregnancy rates after TI were 37.1+/-5.8% for Presync-Ovsynch compared to 35.1+5.0% for Presync-Heatsynch. Use of ECP to induce ovulation was an alternative to GnRH in which greater uterine tone, ease of insemination and occurrence of estrus, improved acceptance by inseminators. A GnRH agonist (Deslorelin; 750 microg) implant inserted at 48 h after injection of PGF(2alpha), as a component of the Ovsynch protocol, induced ovulation, development of a normal CL and delayed follicular growth until 24 d after implant insertion. Utilization of Deslorelin implants (450 microg and 750 microg) to induce ovulation compared to GnRH (100 microg) within the Ovsynch protocol resulted in 27 d pregnancy rates (GnRH 100 microg, 39%; Deslorelin implants 450 microg, 40% and 750 microg, 27.5%) with 12.7%, 5.0% and 9.5% embryonic losses by 41 d of pregnancy, respectively. Induction of an accessory CL with injection of hCG on day 5 after insemination improved conception rates by 7.1%. Bovine somatotrophin injected at first insemination following a Presync-Ovsynch program in cycling-lactating dairy cows increased 74 days pregnancy rates (57.1%>42.6%).     

Ultrasonographic and Endocrine Evaluation of Three Regimes for Oestrus and Ovulation Synchronization for Sheep in the Subtropics

This study aimed to evaluate three regimes for oestrus and ovulation synchronization in Farafra ewes in the subtropics. During autumn, 43 ewes were assigned to (i) controlled internal drug releasing (CIDR)‐eCG group, treated with CIDR for 12 days and eCG at insert withdrawal, n = 13; (ii) PGF₂α‐PGF₂α group, treated with two PGF₂α injections at 11 days interval, n = 14; and (iii) GnRH‐PGF₂α‐GnRH group, treated with GnRH, followed 5 days later with PGF₂α and 24 h later with a second GnRH, n = 16. Oestrus‐mating detection was carried out at 4 h intervals starting on day 0 [the day of CIDR withdrawal (CIDR‐eCG group), the day of second PGF₂α treatment (PGF₂α‐PGF₂α group) and the day of PGF₂α treatment (GnRH‐PGF₂α‐GnRH group)]. Ovarian dynamics was monitored by ultrasound every 12 h beginning on day 0 and continued for 4 days. Blood samples were obtained daily for progesterone (P₄) and oestradiol 17β (E₂) estimation starting on day 0 and continued for 4 days. The obtained results showed that, oestrus expression, ovulation and conception were greater (p < 0.05) in CIDR‐eCG and PGF₂α‐PGF₂α groups than in GnRH‐PGF₂α‐GnRH group. All ewes of PGF₂α‐PGF₂α group presented, on day of second PGF₂α injection with mature CL (P₄ > 2.0 ng/ml), compared to 42.9% in GnRH‐PGF₂α‐GnRH group (p = 0.01). The peak of oestrus occurred 32–52, 48–60 and 28–96 h after the end of treatment in CIDR‐eCG, PGF₂α‐PGF₂α and GnRH‐PGF₂α‐GnRH groups, respectively. Ovulation started 48 h after treatment in all groups and extended for 24, 36 and 48 h for CIDR‐eCG, PGF₂α‐PGF₂α and GnRH‐PGF₂α‐GnRH groups, respectively. Results demonstrated that oestrus and ovulation synchronization could be efficiently achieved in Farafra ewes using either CIDR‐eCG or PGF₂α‐PGF₂α regimes; however, the GnRH‐PGF₂α‐GnRH treatment induced a more spread oestrus and ovulation that may make the protocol inadequate for timed artificial insemination.     

Prostaglandin F2α Stimulates Endothelial Nitric Oxide Synthase Depending on the Existence of Bovine Granulosa Cells: Analysis by Co‐culture System of Endothelial Cells,Smooth Muscle Cells and Granulosa Cells

Prostaglandin F_2α (PGF_2α) induces luteolysis in the mid but not in the early luteal phase; despite this, both the early and the mid corpus luteum (CL) have PGF_2α receptor (FPr). We previously indicated that the luteal blood flow surrounding the CL drastically increases prior to a decrease of progesterone (P) in the cows, suggesting that an acute increase of luteal blood flow may be an early sign of luteolysis in response to PGF_2α and that this may be induced by a vasorelaxant nitric oxide (NO). The aim of this study was to investigate the luteal stage‐dependent and the site‐restricted effect of PGF_2α and NO on the mRNA expressions and P secretion. To mimic the local luteal region both of peripheral and central areas of the CL, we utilized co‐cultures using bovine aorta endothelial cells (EC), smooth muscle cells (SMC) and luteinizing granulosa cells (GC) or fully‐luteinized GC. PGF_2α stimulated the expression of endothelial NO synthase (eNOS) mRNA at 0.5 h in mix‐cultures of EC and SMC with fully‐luteinized GC but not with luteinizing GC. The expression of eNOS mRNA in EC was increased by PGF_2α at 1 h only when EC was cultured together with fully‐luteinized GC but not with luteinizing GC. In all co‐cultures, PGF_2α did not affect the mRNA expression of FPr. Treatment of NO donor inhibited P secretion at 0.5 h. In conclusion, the present study suggests that the coexistence of the mature luteal cells (fully‐luteinized GC) with EC/SMC may be crucial for acquiring functional NO synthesis induced by PGF_2α.     

Timing of Ovulation and Fertility of Heifers After Synchronization of Oestrus with GnRH and Prostaglandin F2α

Synchronization of oestrus and/or ovulation can reduce workload in heifer reproductive management. The objective of this study was to compare two protocols to synchronize oestrus and/or ovulation using GnRH and prostaglandin F_2α (PGF_2α) in dairy heifers concerning their effect on follicular dynamics and reproductive performance. Four trials were carried out. In trial 1, 282 heifers were treated with GnRH and PGF_2α 7 days apart (GP protocol). One group was inseminated on detection of oestrus (IDO 1), and the other group received two timed artificial inseminations (AI) 48 and 72 h after PGF_2α administration (TAI 1). In trial 2, 98 heifers were synchronized with the same GP protocol. Heifers in IDO 2 were treated as in IDO 1, heifers in TAI 2 received two TAI 48 and 78 h after PGF_2α administration. In trial 3, heifers in IDO 3 (n = 71) were again treated as in IDO 1. Heifers in TAI 3 (n = 166) received a second dose of GnRH 48 h after PGF_2α (GPG protocol) and TAI together with this treatment and 24 h later. Trial 4 compared the timing of ovulation after the GP and the GPG protocol, using a subgroup of the heifers from trials 1 to 3. The ovaries of the heifers were scanned via ultrasound at 48, 56, 72, 80, 96 and 104 h after PGF_2α administration. Timing of ovulation and size of the ovulatory follicles were compared between the two groups. In trials 1 to 3, conception rates to first service were between 49 and 66%. They did not differ significantly between IDO and TAI groups within or between trials. Pregnancy rates per synchronization were numerically higher in the TAI groups, but the difference was not significant. Conception rates to breeding on spontaneous oestrus in heifers returning to oestrus were higher than that after synchronized oestrus. In trial 4, more heifers ovulated before the end of the observation period in GPG than in GP (96.5% vs 74.7%; p < 0.001). Overall, ovulatory follicles were smaller in GPG (13.1 ± 1.9 mm vs 14.3 ± 1.9 mm; p < 0.001).     

13,14‐Dihydro‐15‐Keto Prostaglandin F2α Release in Response to Oxytocin Challenge Early Post‐Partum in Anoestrous Nelore Cows Submitted to Temporary Calf Removal and Progesterone Priming

The study evaluated, in early post‐partum anoestrous Nelore cows, if the increase in plasma oestradiol (E2) concentrations in the pre‐ovulatory period and/or progesterone priming (P4 priming) preceding ovulation, induced by hormonal treatment, reduces the endogenous release of prostaglandin PGF₂αand prevents premature lysis of the corpus luteum (CL). Nelore cows were subjected to temporary calf removal for 48 h and divided into two groups: GPE/eCG group (n = 10) and GPG/eCG group (n = 10). Animals of the GPE/eCG group were treated with a GnRH agonist. Seven days later, they received 400 IU of eCG, immediately after PGF₂α treatment, and on day 0, 1.0 mg of oestradiol benzoate (EB). Cows of the GPG/eCG group were similarly treated as those of the GPE/eCG group, except that EB was replaced with a second dose of GnRH. All animals were challenged with oxytocin (OT) 9, 12, 15 and 18 days after EB or GnRH administration and blood samples were collected before and 30 min after OT. Irrespective of the treatments, a decline in P4 concentration on day 18 was observed for cows without P4 priming. However, animals exposed to P4 priming, treated with EB maintained high P4 concentrations (8.8 ± 1.2 ng/ml), whereas there was a decline in P4 on day 18 (2.1 ± 1.0 ng/ml) for cows that received GnRH to induce ovulation (p < 0.01). Production of 13,14‐dihydro‐15‐keto prostaglandin F₂α (PGFM) in response to OT increased between days 9 and 18 (p < 0.01), and this increase tended to be more evident in animals not exposed to P4 priming (p < 0.06). In conclusion, the increase in E2 during the pre‐ovulatory period was not effective in inhibiting PGFM release, which was lower in P4‐primed than in non‐primed animals. Treatment with EB promoted the maintenance of elevated P4 concentrations 18 days after ovulation in P4‐primed animals, indicating a possible beneficial effect of hormone protocols containing EB in animals with P4 priming.     

Combined GnRH and PGF2α Application in Cows with Endometritis Puerperalis Treated with Antibiotics

The investigations were carried out on a total of 70 cows with puerperal endometritis. In addition to intrauterine antibiotic treatment, 30 experimental animals were administered 20 μg GnRH analogue, buserelin, between days 10 and 12 post‐partum followed by 500 μg PGF_2α analogue, cloprostenol, 10 days later. Forty control cows were treated only with intrauterine antibiotics. Blood samples for progesterone determination were collected from the tail vein twice weekly until day 70 post‐partum. The first rise in progesterone level above 3.18 nmol/l occurred significantly earlier in the experimental than in control cows (21.6 ± 9.2 versus 27.8 ± 12.3 days; p ≤ 0.05). The duration of the first cycle post‐partum was 15.0 ± 4.3 days in experimental and 19.7 ± 7.3 days in control animals (p ≤ 0.05). However, no significant differences were observed in the occurrence of first oestrus post‐partum. The involution of the uterus was improved after hormone treatment. At day 42 post‐partum, completion of uterine involution was found in 93.3% of hormone‐treated cows and in 82.5% of those treated with antibiotic only (p ≤ 0.05). Clinical recovery was 96.6% in the experimental and 82.5% in the control group (p ≤ 0.05). First service pregnancy rate was significantly better in hormone‐treated than control cows (51.7 versus 36.4%; p ≤ 0.05). Total pregnancy rate and insemination index values were not significantly improved following GnRH and PGF_2α treatment. The average service period was 89.8 ± 21.2 days in cows after hormone treatment, and 112.6 ± 24.5 days in control cows. The difference was statistically significant (p ≤ 0.05). These results indicate, that the sequential GnRH and PGF_2α application in cows with puerperal endometritis positively affected ovarian function and uterine involution, resulting in improved fertility performance.     

Effect of ovulatory follicle size and expression of estrus on progesterone secretion in beef cows

Induced ovulation of small dominant follicles (SF, < 12 mm; CO-Synch protocol) in postpartum beef cows resulted in formation of corpora lutea (CL) that exhibited a delayed rise in progesterone (P4) compared with CL from large dominant follicles (LF, > 12 mm). Experiment 1 characterized P4 concentrations from ovulation to subsequent estrus among GnRH-induced or spontaneously ovulated SF (or= 12 mm) to determine if P4 secretion by CL formed from GnRH-induced SF remains lower postovulation in nonlactating beef cows. Nonlactating beef cows were induced to ovulate 48 h after PGF(2alpha) (CO-Synch; GnRH on d - 9, PGF(2alpha) on d - 2, and GnRH on d 0) or exhibited estrus and spontaneously ovulated after PGF(2alpha). Follicle size was measured at the second GnRH in cows induced to ovulate or approximately 3 h after the onset of estrus for cows that ovulated spontaneously. Cows were classified into 1 of 4 groups: 1) GnRH-induced ovulation-SF (or= 12 mm; Ind-LF; n = 16); 3) spontaneous ovulation-SF (or= 12 mm; Spon-LF; n = 22). Serum concentrations of P4 from d 3 to 15 were reduced in the Ind-SF compared with the Ind-LF (P = 0.05), Spon-SF (P = 0.07), and Spon-LF (P = 0.03). Experiment 2 characterized P4 concentrations (0 to 60 d postAI) among GnRH-induced or spontaneously ovulated SF (or= 13 mm) to determine if P4 secretion by CL formed from GnRH-induced SF remained lower during early gestation. Ovulation was induced with GnRH 48 h after PGF(2) (CO-Synch) or occurred spontaneously, and ovulatory follicle size was measured at AI. Lactating cows were classified into 1 of 3 groups: 1) GnRH-induced ovulation-SF (or= 13 mm; Ind-LF; n = 43); or 3) spontaneous ovulation-LF (>or= 13 mm; Spon-LF; n = 27). The increase in P4 concentrations was greater (P = 0.06) in pregnant (d 2 to 12) compared with nonpregnant cows. Also, the increase in P4 from d 2 to 12 was greater (P = 0.01) in the Ind-LF compared with the Ind-SF groups, but there was no difference (P = 0.94) among groups in P4 from d 14 to 60 in pregnant cows. Follicle size at AI influenced the increase in P4 in cows that failed to conceive (P = 0.007), but not among cows that became pregnant (P = 0.32) to AI. In summary, P4 secretion after GnRH-induced ovulation of SF was decreased from d 2 to 12 compared with that of LF, but was similar among pregnant cows from d 14 to 60 postAI (d 0).     

Luteolytic Effect of Prostaglandin F2α on Bovine Corpus Luteum Depends on Cell Composition and Contact

Prostaglandin F_2α (PGF_2α) is a main luteolytic factor in vivo; however, its direct luteolytic influence on steroidogenic cells of bovine corpus luteum (CL) is controversial and not fully understood. The aim of the study was to clarify PGF_2α action on bovine CL in different in vivo and in vitro conditions and to examine whether the contact among all main types of CL cells is necessary for luteolytic PGF_2α action. In experiment 1, the bovine CL (day 15 of the oestrous cycle) was perfused using in vivo microdialysis system with dinoprost (an analogue of PGF_2α) for 0.5 h. Dinoprost caused a short‐time increase in progesterone (P4), whose concentration decreased thereafter (at 6‐, 10‐, 12‐ and 24‐h after treatment). In experiment 2, the direct effect of PGF_2α on P4 accumulation in CL steroidogenic cells cultured in monolayer (day 15 of the cycle) was determined. PGF_2α after 24 h of incubation increased P4 accumulation in steroidogenic CL cells. In experiment 3 steroidogenic, endothelial CL and immune cells (day 15 of the cycle) were incubated with PGF_2α in cocultures for 24 h in glass tubes and the levels of P4, stable metabolites of nitric oxide (NO) and leukotriene (LT) C₄ were determined. Although PGF_2α treatment increased P4 secretion in homogeneous steroidogenic CL cell culture, the decrease in P4 secretion in cocultures of all types of CL cells was observed. The secretion of NO and LTC₄ increased after the treatment of PGF_2α both in pure cultures of CL cells and in cocultures. The interactions between endothelial and immune cells with steroidogenic CL cells are needed for luteolytic PGF_2α action within the bovine CL. Our results indicate that the cell coculture model, including the main types of CL cells, is the most approximate to study PGF_2α role in vitro.     

Induction of ovulation with GnRH and PGF2alpha in lactating Bos taurus x Bos indicus cows

Induction of ovulation for timed artificial insemination (TAI) with the Ovsynch protocol was evaluated in 49 anoestrous and lactating Bos taurus x Bos indicus cows. Palpation per rectum and transrectal ultrasonography were used on Days -30, -20, -10 and 0 (start of treatment) to confirm anoestrus but with the presence of follicles > or = 10 mm, and every other day during treatment to determine ovarian activity. Cows were randomly assigned to: (1) Ovsynch (n = 24; Day 0, 200 microg GnRH; Day 7, 150 microg PGF2alpha; Day 9, 200 microg GnRH + TAI 16 to 20 h later) and (2) control (n = 25; no treatment). Rates of ovulation for the first GnRH injection, detection of a corpus luteum (CL) at PGF2alpha injection, pregnancy and induction of cyclicity were greater (P < 0.05) with Ovsynch. There was no effect of body condition score (P > 0.05). In conclusion, the Ovsynch protocol was not effective in obtaining acceptable pregnancy rate for TAI, but it was effective for induction of cyclicity in anoestrous and lactating Bos taurus x Bos indicus cows under tropical conditions.     

Regulation of uterine function by cytokines in cows: Possible actions of tumor necrosis factor-α, interleukin-1α and interferon-τ

When animals do not become pregnant, regression of the corpus luteum (CL) is essential for normal cyclicity because it allows the development of a new ovulatory follicle. Luteal regression is caused by a pulsatile release of prostaglandin (PG) F_2α from the uterus in the late luteal phase in most mammals including cattle. Although it has been proposed in ruminants that pulsatile PGF_2α secretion is generated by a positive feedback loop between luteal and/or hypophyseal oxytocin and uterine PGF_2α, the bovine endometrium may possess other mechanisms for initiation of luteolytic PGF_2α secretion. There is increasing evidence that several cytokines mainly produced by immune cells modulate CL and uterine function in many species. Tumor necrosis factor‐α (TNF‐α) stimulates PGF_2α output from bovine endometrium not only at the follicular phase but also at the late luteal phase. Administration of TNF‐α at a high concentration prolongs luteal lifespan, whereas administration of a low concentration of TNF‐α accelerates luteal regression in cows. The data obtained from the authors’ previous in vitro and in vivo studies strongly suggest that TNF‐α is a crucial factor in regulating luteolysis in cows. The authors’ recent study has shown that interleukin‐1α mediates PG secretion from bovine endometrium as a local regulator. Furthermore, interferon‐τ (IFN‐τ) suppresses the action of TNF‐α on PGF_2α synthesis by the bovine endometrium in vitro, suggesting that IFN‐τ plays a luteoprotective role by inhibiting TNF‐α‐induced PGF_2α production in early pregnancy. The purpose of the present review is to summarize current understanding of the endocrine mechanisms that regulate uterine function by cytokines during the estrous cycle and early pregnancy in cows.     

Presence of multiple corpora lutea affects the luteolytic response to prostaglandin F2α in lactating dairy cows

Since the 1970s, luteolytic doses used for synchronizing estrus in dairy cattle have remained unchanged. This study aimed to evaluate the dose-response effect of prostaglandin F_2α (PGF_2α), which is used for synchronizing estrus, and subsequent fertility in cows with two or more corpora lutea (CL). The study population consisted of 1,683 cows with a single CL (1CL), 501 cows with multiple CL receiving a single dose of PGF_2α (2CL1), and 252 cows with multiple CL receiving a 1.5 × PGF_2α dose (2CL1.5). Cows with a single CL (n = 1,245) showed estrus significantly (P < 0.01) earlier (3.01 ± 1.23 days; mean ± SD) than cows with multiple CL (n = 287; 3.33 ± 1.69 days). Using 1CL cows as reference, the odds ratio (OR) for the estrus response in 2CL1 cows was 0.13 (P < 0.0001), whereas the ORs for estrus response and pregnancy of 2CL1.5 cows were 1.8 (P = 0.0001) and 1.7 (P = 0.001), respectively. Based on the results for only the 2CL1 cows, the OR for the estrus response was 0.7 (P = 0.01) for cows producing ≥ 45 kg of milk at treatment, compared to the remaining cows producing < 45 kg of milk. Our results showed that the presence of multiple CL reduced the estrus response to that induced by a single PGF_2α dose and milk production was inversely associated with this response, whereas an increased PGF_2α dose improved the estrus response. Therefore, an increase in the standard PGF_2α dose is recommended.     

Effect of estradiol cypionate and GnRH treatment on plasma estradiol‐17β concentrations,synchronization of ovulation and on pregnancy rates in suckled beef cows treated with FTAI‐based protocols

Two experiments were conducted to evaluate the effect of different ovulation inducers on E‐17β plasma concentrations, synchronized ovulations and pregnancy rates. In Experiment 1, cows received a progesterone intravaginal device (PID) with 1 g of progesterone (P4) plus 2 mg of estradiol benzoate (EB) (day 0). At PID removal (day 8), cows received 0.150 mg of D‐cloprostenol and were randomly assigned to four treatment groups (n = 10/treatment): Group ECP: 1 mg of estradiol cypionate at PID removal, Group EB: 1 mg of EB 24 hr after PID removal, Group GnRH: 10 μg of GnRH 48 hr after PID removal, Group ECP‐GnRH: 1 mg of ECP at PID removal plus 10 μg of GnRH 48 hr later. Ultrasonographic examinations were performed to detect the dominant follicle and ovulation. GnRH‐treated cows ovulated later (p < .05) compared to ECP‐ and ECP+GnRH‐treated cows. There were effects of treatment, time and their interaction on E‐17β concentrations (p < .05). ECP treatment affected plasma E‐17β concentration, which increased earlier and decreased later compared to treatments without ECP. In Experiment 2, cows received (i) ECP: n = 126; (ii) EB: n = 126; (iii) GnRH: n = 136; (iv) ECP+GnRH: n = 139; FTAI was performed 48–50 hr after PID removal. Pregnancy rates did not differ among ovulation inducers (p > .05; ECP: 54.0%, 68/126; EB: 49.2%, 62/126; GnRH: 40.4%, 55/136; ECP+GnRH: 43.9%, 61/139). In conclusion, ECP administration (ECP and ECP+GnRH treatments) affected E‐17β concentrations, determining its earlier increase and later decrease compared to treatments without ECP (EB and GnRH treatments). ECP+GnRH‐treated cows achieved the best distribution of ovulations without affecting pregnancy rates.     

Effect of Progesterone Prior to GnRH-PGF2α Treatment on Induction of Oestrus and Pregnancy in Anoestrous Awassi Ewes

An experiment was conducted to examine the effect of progesterone prior to a GnRH‐PGF_2α treatment on oestrus and pregnancy in seasonally anoestrous Awassi ewes. Twenty‐four ewes were randomly assigned to three groups to be pre‐treated with 60 mg medroxyprogesterone acetate sponges (group A), 600 mg progesterone sponges (group B) or blank sponges (group C) for 4 days. All ewes were injected with 100 μg of GnRH 24 h after sponge removal followed, 5 days later, by 20 mg PGF_2α injection. Ewes were exposed to three fertile rams at the time of PGF_2α injection (day 0, 0 h) and were checked for breeding marks at 6‐h intervals for 5 days. Blood samples were collected from all ewes 1 day (day ?10) prior to sponge insertion, at the time of sponge removal (day ?6), 1 day following sponge removal (day ?5, at the time of GnRH injection) and at the time of PGF_2α injection (day 0) for analysis of progesterone. Progesterone concentrations on days ?10 and ?5 were basal and averaged 0.2 ± 0.04 and 0.2 ± 0.2 ng/ml, respectively. Progesterone concentrations on day ?6 were elevated only in group B ewes and were higher (p < 0.0001) than those of groups A and C. Progesterone concentrations on day 0 were higher (p = 0.002) in groups A and B than group C. Oestrous responses occurred only in ewes of groups A and B (p > 0.05). Induced oestrus conception rate was greater (p < 0.01) in group A than groups B and C. Ewes returned to oestrus 17–20 days following day 0 were two of eight, six of eight and three of eight of groups A, B and C, respectively, all of which eventually lambed. The overall lambing rate was 82% in progesterone‐primed ewes compared with only 38% non‐progesterone‐primed ewes (p < 0.05). Progesterone priming apparently sensitizes GnRH‐PGF_2α‐treated seasonally anoestrous ewes and increases their response in oestrus and pregnancy rates.     

Evaluation of the effect of a 3rd GnRH injection administered six days after the 2nd GnRH injection of Ovsynch on the reproductive performance of Japanese black cows

This study was designed to evaluate the reproductive performance of Japanese black cows following the 3rd injection of gonadotropin releasing hormone (GnRH) analogue administered concurrently with Ovsynch-based treatment on day 6 (day 1 = the day of ovulation). In Experiment 1, 12 cows were allocated into three groups: a control group that was subjected to Ovsynch treatment and then injected with a placebo on day 6; group 1 (Ovsynch + GnRH), which was subjected to Ovsynch treatment and was injected with GnRH analogue on day 6, and group 2 (Ovsynch + controlled internal drug-release (CIDR) + GnRH), which received Ovsynch-CIDR treatment and was injected with GnRH analogue on day 6. Blood collection and ultrasonographic observation of the ovaries were conducted daily. Both treatments induced the formation of an accessory corpus luteum and significantly increased the cross-sectional area of the luteal tissue when compared to the control. However, plasma progesterone (P₄) was significantly higher in the treatment groups than in the control group on days 11, 12, 17 and 18 in the group 1 and from day 10 to 21 in the group 2. In Experiment 2, 41 cows were assigned to the same three groups described above and then artificially inseminated on day 1. The pregnancy rates on day 45 did not differ among groups. In conclusion, administration of GnRH analogue on day 6 following Ovsynch-based treatment did not improve the reproductive performance of Japanese black cows, even though the P₄ concentration was higher in groups that received the GnRH.