In vivo and in vitro studies of the role of the adrenergic system and follicular wall contractility in the pathogenesis and resolution of bovine follicular cysts

Bovine follicular cysts (FCs) are a common cause of economic loss in modern dairy herds. Their aetiopathogenesis is not completely understood, even though an inadequate hypothalamic release of GnRH at the time of ovulation is considered to be their main cause. Much evidence, however, suggests a role for adrenergic innervation in ovarian functions, such as follicular development, steroid hormone secretion, and follicular contractility, the latter being an event important for ovulation. Moreover, in humans, polycystic ovary syndrome, a disease very similar to bovine follicular cysts, is characterised by increased density of adrenergic nerves. Given these premises, the aim of our study was to evaluate the effectiveness and mode of action of a novel strategy for the treatment of bovine follicular cysts. In the in vivo experiment, 170 Friesian cows diagnosed with follicular cysts were assigned to four groups (groups A, B, C, and D) to assess the effects of epidural administration of a β-adrenergic antagonist (carazolol) alone or in combination with a GnRH analogue (lecirelin). The four groups underwent the following treatments: Group A was administered lecirelin in combination with carazolol; Group B was administered carazolol; Group C was administered lecirelin; and Group D was administered only normal saline solution. In the in vitro experiment, strips of the walls of cystic follicles recovered post-mortem were suspended in an organ bath, connected to an isometric force transducer and exposed to increasing doses of epinephrine or to the same treatment after exposure to carazolol for 15 min (n = 10). The amplitude and frequency of the contractile activity were recorded. None of the control cows was observed in oestrus or was submitted to AI. The combination of lecirelin and carazolol induced a significant increase in the number of cows in oestrus (88%) compared to lecirelin alone or to carazolol alone (P < 0.05 and P < 0.01, respectively). The combination of lecirelin and carazolol and lecirelin alone were significantly more efficacious than carazolol alone (P < 0.01 and P < 0.05, respectively). In the in vitro experiment, epinephrine increased the amplitude of the contractions of the strips in a dose-dependent manner. This response was significantly enhanced in strips pre-treated with carazolol. The treatments had no effect on the frequency of contractions. In conclusion, our work demonstrates that the epidural administration of a GnRH analogue and a β-adrenergic blocker is an effective means of treating cows with cystic ovarian disease. Moreover, it confirms, from a clinical point of view, that alterations of the adrenergic system and of the contractility of the follicular wall can be considered aetiopathogenic factors involved in the development of FCs. The results of this study lay the basis for a new therapeutic approach to FCs.     

Renin and ovarian vascularization in cows with follicular cysts after epidural administration of a GnRH analogue

The ovarian renin–angiotensin system may play an important role in follicular growth and maturation, as well as in the process of ovulation. The aim of this study was to investigate the effects of administration of a GnRH analogue to cows with ovarian follicular cysts on plasma renin concentrations and ovarian vascularization. This study was performed with 60 Friesian cows, which were diagnosed with follicular cysts, and randomly allocated into two groups: group A (treatment; n = 30) received 2 ml of lecirelin (Dalmarelin^® – Fatro), per head via sacro-coccygeal epidural, and group B (control; n = 30) received 2 ml saline solution (0.9% NaCl) per head by the same route. Blood samples were immediately collected prior to administration (T0) and then 24 h (T1), 48 h (T2) and 8 days (T3) after administration of the treatment, for both groups. Ovarian vascularization was evaluated utilizing Power Doppler on these same days in 10 animals from each group. The number of pixels detected by Power Doppler was used as an indicator of the degree of vascularization. Plasma renin concentrations remained relatively constant for the control (group B) animals, but increased as the sampling period progressed (NS) for the treated cows (group A). Similarly, there were no changes in ovarian vascularization (number of pixels) for the control cows, but vascularization increased throughout the sampling period in the treated animals. The number of pixels associated with cysts was significantly higher for treated compared to control cows at 24 h after treatment (P < 0.001). The epidural administration of a GnRH analogue was determined to be a highly effective therapy for follicular cysts (regression occurred in 82% of treated cows within 8 ± 2 days after treatment, but in none of the control cows), which also enhanced ovarian vascularization.     

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 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.     

Effectiveness of prostaglandin F2alpha in the initial treatment of bovine ovarian cysts

This study was conducted to evaluate the use of prostaglandin F2alpha (PGF2alpha) in the initial treatment of ovarian cysts in dairy cattle. Two hundred and sixty three cows diagnosed cystic on palpation per rectum were randomly assigned to one of three treatment groups (A, B or C). Cows in Groups A and B were treated with 25 mg i.m.of PGF2alpha at the time of diagnosis (Day 0), while cows in Group C received 100 mug of GnRH. Seven days following initial treatment (Day 7), cows from Group A that were not observed in estrus were treated with GnRH. Cows from Groups B and C were not treated. On Day 14, all cows that had not been inseminated received PGF2alpha. A blood sample was obtained from all cows on Days 0, 7 and 14 and was analyzed for progesterone (P4) using radioimmunoassay. Incidences of estrus were recorded and cows that were more than 60 d in milk at the time of diagnosis were bred when observed in estrus. The incidence of follicular cysts on Day 0 (as defined as P4 <0.5 ng/ml) was similar between groups and constituted about 40% of all cysts. There were significantly more cows pregnant to insemination within 7 d of initial treatment in Group B than in Groups A and C (P<0.05). After Day 14, the pregnancy rate was not statistically different between Group B and C, but Groups B and C had a statistically higher pregnancy rate than Group A from Day 21 to Day 35. At the end of the study, there was no statistical difference for the pregnancy rate between groups. We concluded that treatment of ovarian cysts diagnosed by per rectum examination with prostaglandin (at time of diagnosis and 14 d later for cows that were not inseminated) was as effective as initial treatment with GnRH followed by prostaglandins 14 d later for cows that were not inseminated previously. Cows that were initially treated with prostaglandins also tended to become pregnant sooner.     

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.     

Bovine ovarian follicular cysts: in vitro effects of lecirelin, a GnRH analogue

This study investigates the mechanisms of action by which a GnRH analogue may modulate the contractility of the bovine ovarian follicular wall. The in vitro evaluation of the spontaneous basal contractility of bovine preovulatory and cystic follicles was performed, followed by testing the effects of lecirelin, a GnRH analogue, on their basal contractility. Strips of tissue in isolated organ bath were employed.In addition, to better investigate the mechanism of action of lecirelin, the study of the effects of cumulative doses of nifedipine (a calcium channel blocker), phentolamine (an α-adrenoceptor antagonist) and reserpine (an inhibitor of the vesicular up-take of catecholamines) alone and, at the highest doses employed, associated to lecirelin, was set up.The results demonstrate that in basal conditions and after the addition of lecirelin, the strips from preovulatory follicles contract significantly more than strips from cysts.Furthermore, among the patterns of contractility evoked by the three drugs employed, the one induced by nifedipine was the only one unaffected by the addition of lecirelin.The data obtained provide the hypothesis that one of the main mechanisms of action of GnRH, could involve calcium channels.     

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

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

The effect of interval from day of administration of bovine somatotropin (bST) to synchronization of ovulation and timed-insemination on conception rate of dairy cows with and without ovarian cysts

The objective of this study is to evaluate the effect of the interval from the day of administration of bovine somatotropin (bST) to the day of initiation of synchronization of ovulation (Day 0 and 7) and timed-insemination (TAI) on conception rate (CR) of dairy cows with and without ovarian cysts, respectively. Lactating dairy cows (n = 359) were divided into two groups. Cows in Group 1 (n = 238, without ovarian cysts) were treated with 100 microg, i.m. GnRH on Day 0; 25 mg, i.m. PGF2a on Day 8. 100 mirog. i.m. GnRH on Day 10; and inseminated 16 h later without detection of estrus. Cows in Group 2 (n = 121, with ovarian cysts) were treated with 100 microg, i.m. GnRH on Day 0; 100 microg, i.m. GnRH on Day 7; 25 mg, i.m. PGF2alpha on Day 14, 100 ug, i.m. GnRH on Day 16; and inseminated 16 h later without detection of estrus. Between 60 and 63 days postpartum, all cows in the herd were given bST every 14 days for the duration of the study. Logistic regression was used to assess the risk of nonpregnancy associated with interval from bST treatment to Day 0 for cows without ovarian cysts. and both Day 0 and 7 for cows with ovarian cysts adjusting for parity and days in milk. The CR for cows in Group 1 was significantly higher when the interval from last treatment with bST to Day 0 was between 1 and 3 days (28%) compared to 4-6 days (14%). In addition, the risk of nonpregnancy was 2.19 times greater in cows 4-6 days after bST treatment compared to 1-3 days after adjusting for parity and days in milk. The CR for cows in Group 2 was not significantly different when the interval from last treatment with bST to both Day 0 and 7 was between 1 and 3, 4 and 6, and 7 and 14 days. In conclusion, the results of this study suggested bST treatment closer to Day 0 had a positive effect on CR of cows without ovarian cysts, but bST treatment closer to both Day 0 and 7 had no effect on CR of cows with ovarian cysts. This was interpreted to mean that bST had a beneficial effect on either, or both, the preovulatory follicle and the oocyte in dairy cows without ovarian cysts, but not in dairy cows with ovarian cysts.     

Resynchronization of ovulation and timed insemination in lactating dairy cows, II: assigning protocols according to stages of the estrous cycle, or presence of ovarian cysts or anestrus

Pregnancy rates were compared in lactating dairy cows (n = 1083) assigned to protocols for resynchronization of ovulation based on stages of the estrous cycle, or presence of ovarian cysts or anestrus. Cows were detected not pregnant by ultrasonography 30 d after a previous AI (study day 0) and classified as diestrus, metestrus, proestrus, with ovarian cysts or anestrus. Cows in diestrus (January-May) were assigned to either Ovsynch (GnRH day 0, PGF2alpha day 7, GnRH day 9, and timed-AI [TAI] 16 h later; n = 96), or Quicksynch (PGF2alpha day 0, estradiol cypionate [ECP] day 1, AI at detected estrus [AIDE] on day 2, or TAI on day 3; n = 96). Cows in diestrus (June-December) were assigned to either Ovsynch (n = 156) or Modified Quicksynch (PGF2alpha day 0, ECP day 1, AIDE days 2 and 3, and to Ovsynch on day 4 if not detected in estrus; n = 142). Cows in metestrus were assigned either to Ovsynch (n = 68), Heatsynch (GnRH day 0, PGF2alpha day 7, ECP day 8, AIDE day 9, or TAI day 10; n = 62), or GnRH + Ovsynch (GnRH on day 0, followed by Ovsynch on day 8; n = 64). Cows in proestrus, with ovarian cysts, or anestrus were assigned to either Ovsynch (proestrus n = 89, ovarian cysts n = 97, anestrus n = 8) or GnRH + Ovsynch (proestrus n = 87, ovarian cysts n = 109, anestrus n = 9). Pregnancy rate was evaluated 30, 55 and 90 d after resynchronized AI. For cows in diestrus (January-May), pregnancy rates were higher for Ovsynch (35.9, 29.2 and 26.0%) than for Quicksynch (21.7, 16.7 and 15.6%). For cows in diestrus (June-December), pregnancy rates were similar for Ovsynch (34.4, 24.0 and 23.6%) and Modified Quicksynch (27.1, 26.2 and 21.6%). For cows in metestrus, pregnancy rates were higher for GnRH + Ovsynch (33.3, 24.5 and 20.3%) than for Heatsynch (20.3, 12.9 and 9.8%). For cows with ovarian cysts, pregnancy rates were higher for GnRH + Ovsynch (30.3, 26.6 and 22.9%) than for Ovsynch (20.2, 18.5 and 14.7%). Assignment to resynchronization protocols based on the stages of the estrous cycle, or presence of ovarian cysts improved pregnancy rates.     

Hormone response of dairy cows with ovarian cysts after treatment with HCG or GnRH

Twenty lactating Holstein and Guernsey cows, diagnosed by rectal palpation as having ovarian cysts, were randomly divided within breed into two groups to receive either a single intramuscular injection of 100 μg of synthetic gonadotropin releasing hormone (GnRH) or an intravenous injection of 10,000 IU of human chorionic gonadotropin (HCG). The objective was to compare hormonal and clinical changes in cows with ovarian cysts following treatment with GnRH and HCG. Eight of ten and nine of ten cows given either GnRH and HCG, respectively, responded to treatment and subsequent fertility was not different between the two groups. Pre-injection plasma levels of LH, progesterone, and estradiol were highly variable. Mean plasma levels of LH, progesterone and estradiol did not differ between groups either following treatment (days 1–17 post-treatment), at the subsequent estrus, or during days 1–13 following the subsequent estrus. Mean LH levels did not differ significantly on the days either post-treatment or post-estrus except that levels were higher (P < .01) at the subsequent estrus as compared to the other days. Mean progesterone levels increased after treatment with either GnRH or HCG and were higher on days 5, 9 and 13 post-estrus and post-treatment as compared to the subsequent estrus. Mean levels of estradiol were higher (P < .05) at the subsequent estrus than any other time post-treatment or post-estrus. No other days were significantly different. In conclusion, GnRH and HCG are effective treatments for ovarian cysts in cattle. Endocrine response on days following treatment are similar for both compounds.     

Importance of estrus on pregnancy per insemination in suckled Bos indicus cows submitted to estradiol/progesterone-based timed insemination protocols

The objective was to evaluate the effect of estrus occurrence (based on removal of tail-head marks) on ovarian responses and pregnancy per AI (P/AI; 30 d after AI) in suckled Bos indicus beef cows submitted to timed AI (TAI) protocols. Cows received an intravaginal device containing 1.0 g progesterone, and 2.0 mg estradiol benzoate im; 8 d later, the intravaginal device was removed, and they were given PGF_2α (0.25 mg of cloprostenol sodium) and 300 IU of eCG, with TAI 48 to 52 h later. In Experiment 1, cows were assigned to receive one of three treatments: 1 mg of estradiol cypionate (ECP) im at progesterone (P4) device removal (N = 178); 10 μg of GnRH im at TAI (N = 190); or both treatments (N = 172). In cows given estradiol (ECP or ECP + GnRH), more displayed estrus (P = 0.002) and became pregnant (P < 0.0001) compared with those receiving only GnRH. In Experiment 2, the effect of the occurrence of estrus on ovarian responses was evaluated in cows (N = 53) synchronized using ECP at device removal. Cows that displayed estrus had a greater diameter of the largest follicle (LF) at device removal (P < 0.0001), a greater diameter at TAI (P < 0.0001), a greater ovulation rate (P = 0.02), a larger CL (P = 0.02), and a greater P4 concentration (P < 0.0001) than cows that did not display estrus. In Experiment 3, the effect of GnRH treatment on P/AI at TAI was evaluated in cows that received ECP at device removal, and either displayed, or did not display, estrus (N = 726). There was no estrus by GnRH interaction (P = 0.22); the P/AI was greater (P < 0.0001) in cows that displayed estrus (61.9%) than cows that did not display estrus (41.4%). However, GnRH did not improve (P = 0.81) P/AI (GnRH = 53.7% vs. no GnRH = 52.6%). In conclusion, exogenous estradiol at device removal increased both the proportion of suckled Bos indicus cows that displayed estrus and P/AI. Cows that displayed estrus had better ovarian responses (i.e., larger follicles at TAI, a greater ovulation rate, larger CL, and greater P4 concentrations) following an estradiol/P4-based synchronization protocol. Although occurrence of estrus improved pregnancy outcomes, GnRH at TAI did not improve P/AI in suckled Bos indicus cows treated with ECP, regardless of estrus occurrence.     

Influence of intrauterine infections and follicular development on the response to GnRH administration in postpartum dairy cows

The effects of intrauterine infections and prior follicular development on the response to gonadotropin releasing hormone (GnRH) administration in postpartum dairy cows were studied. Fifty lactating Holstein cows were assigned at random to one of two groups after calving. Group I (control) consisted of 25 cows given a single intramuscular injection of saline on Day 15 postpartum. Group II (treated) consisted of 25 herdmates given a single i.m. injection of 100 mug of GnRH on Day 15 postpartum. Palpation per rectum and real-time ultrasonography were used to monitor ovarian activity, and endometrial swabs were cultured to determine the presence of uterine infection. Blood samples were collected for progesterone (P(4)) and luteinizing hormone (LH) analysis. Fourteen cows (control, n = 5; treated, n = 9) did not ovulate during the first 60 d postpartum. Ovaries in these cows contained 4 to 8-mm size follicles and both P(4) and LH remained at basal concentrations. Fourteen other cows (control, n = 6; treated, n = 8) ovulated by Day 15 postpartum. Follicles >/= 10 mm were demonstrable in the ovaries of these cows before or by Day 12 postpartum. GnRH treatment had no effect on the lifespan of the existing corpus luteum in these cows. In the remaining cows, 7 of 14 Control and all 8 Treated cows ovulated within 3 d of treatment. All cows ovulating within this period were free of uterine infection and the ovaries contained follicles 相似文献   

A CIDR-based timed AI protocol can be effectively used for dairy cows with follicular cysts

The present study evaluated whether a controlled internal drug release (CIDR)-based timed AI (TAI) protocol could be used as an efficient tool for the treatment of ovarian follicular cysts in lactating dairy cows. In the first experiment, lactating dairy cows diagnosed with follicular cysts were randomly assigned to two treatments: (1) a single injection of GnRH at diagnosis (Day 0) and AI at estrus (AIE) within 21 days (GnRH group, n=70), or (2) insertion of a CIDR device containing progesterone and an injection of GnRH on Day 0, PGF(2alpha) injection at the time of CIDR removal on Day 7, GnRH injection on Day 9, and TAI 16h after the GnRH injection (CIDR-based TAI group, n=65). Conception rate after the CIDR-based TAI protocol (52.3%) was greater (P<0.05) than that after AIE following a single GnRH injection (26.9%). In the second experiment, lactating dairy cows diagnosed with follicular cysts (Cyst group, n=16) and cows having normal estrous cycles (CYC group, n=15) received the same treatment: a CIDR device containing progesterone and an injection of GnRH on Day 0, PGF(2alpha) injection at the time of CIDR removal on Day 7, and GnRH injection on Day 9. The proportion of cows with follicular wave emergence and the interval from treatment to follicular wave emergence did not differ (P>0.05) between groups. The mean diameters of dominant follicles on Days 4 and 7 as well as preovulatory follicles on Day 9, and the synchrony of ovulation following the second injection of GnRH did not differ (P>0.05) between groups. These data suggest that the CIDR-based TAI protocol results in an acceptable conception rate in dairy cows with follicular cysts.     

Resynchronization of ovulation and timed insemination in lactating dairy cows I: use of the Ovsynch and Heatsynch protocols after non-pregnancy diagnosis by ultrasonography

The objective was to compare pregnancy rates and pregnancy losses in lactating dairy cows that were diagnosed not pregnant and re-inseminated following either the Ovsynch or Heatsynch protocols. Also evaluated were the effects of stages of the estrous cycle, ovarian cysts and anestrus on pregnancy rates for both treatments. Non-pregnant cows (n = 332) as determined by ultrasonography on day 27 post-AI (study day 0) were divided into two groups. Cows in the Ovsynch group (n = 166) received GnRH on day 0, PGF2alpha on day 7, GnRH on day 9, and timed AI (TAI) 16 h later (day 10). Cows in the Heatsynch group (n = 166) received GnRH on day 0, PGF2alpha on day 7, estradiol cypionate (ECP) on day 8, and TAI 48 h later (day 10). Cows detected in estrus on days 8 and 9 were inseminated and included in the study. On day 0, cows were classified according to different stages of the estrous cycle, or presence of ovarian cysts or anestrus. Pregnancy rates were evaluated 27, 45 and 90 days after resynchronized AI. Overall, there was no difference in pregnancy rates on days 27, 45 and 90 between cows in the Ovsynch (25.2, 17.5, and 13.9%) and Heatsynch (25.8, 19.9, and 16.1%) groups. There was no difference in pregnancy losses from days 27 to 45 and days 45 to 90 for cows in the Ovsynch (25.0 and 17.9%) and Heatsynch (14.7 and 10.3%) groups. However, pregnancy rates were increased when cows in metestrus were subjected to the Heatsynch protocol and cows with ovarian cysts were subjected to the Ovsynch protocol.     

The effect of a GnRH analogue on the dynamics of follicular development and synchronization of estrus in lactating cyclic dairy cows

A GnRH analogue was used to synchronize ovarian follicular development prior to an injection of PGF(2alpha) for the synchronization of estrus in lactating Holstein cows. On Day 12 (estrus = Day 0) of the experimental cycle, cows (n = 8) were injected with 8 mug Buserelin (BUS group), followed by 25 mg PGF(2alpha) 7 d later (Day 19). Control cows (n = 7) received PGF(2alpha) on Day 12 (PGF group). Ovaries were scanned daily via ultrasonography, and plasma progesterone and estradiol concentrations were determined. Sizes of all visible follicles were recorded. Follicles were classified as small (3 to 5 mm), medium (6 to 9 mm), or large (>/= 10 mm). Between Days 12 and 16 of the cycle, the number of large follicles in PGF cows remained unchanged (1.2), whereas in the BUS group, the number of large follicles decreased from 1.3 on Day 12 to 0.5 on Day 15. Only 4 of 7 PGF cows ovulated a second-wave dominant follicle. In the BUS group, 7 of 8 cows ovulated a GnRH analogue induced dominant follicle that was first identified on Day 15. During the follicular phase (last 5 d prior to estrus), plasma progesterone declined in association with CL regression in both groups, and estradiol concentrations increased, reaching higher (P<.0.05) preovulatory peak concentration in BUS cows than in PGF cows (14.0 +/- 1.0 vs 10.4 +/- 1.1 pg/ml). The number of medium-size follicles was smaller and the number of small-size follicles tended to be higher in BUS cows than in the PGF-treated group. On the day of estrus, the size of the ovulatory follicle (16.1 vs 13.3 mm) and the size difference between the ovulatory and second largest follicle (11.4 vs 6.2 mm) were both larger in BUS cows than in PGF-treated cows, suggesting a more potent dominance effect of the ovulatory follicle in the BUS cows. This study suggests that a GnRH analogue can alter follicular development prior to synchronization of estrus with an injection of PGF(2alpha) in lactating dairy cows.     

Effect of gonadotropin-releasing hormone and prostaglandin F2α on reproduction in postpartum dairy cows

Thirty dairy cows serving as the treated group (Group A) were injected intramuscularly with 100 mcg gonadotropin-releasing hormone (GnRH) at 10 to 16 days postpartum followed by 25 mg prostaglandin F₂α (PGF₂α) 14 days later. Twenty-nine herdmate dairy cows (Group B) serving as controls were treated in a similar manner using saline injections rather than GnRH or PGF₂α treatments. Only cows without obvious uterine infection were assigned to the experimental groups, and any uterine pathology that developed during the treatment interval was treated accordingly following the experimental period. Internal genitalia were evaluated via rectal palpation prior to each injection. Blood samples were collected for progesterone analysis before each injection and at 30 hours following the PGF₂α or the second saline injection. Experimental animals were artificially inseminated at the first detected postpartum estrus starting 35 to 40 days following calving. Results indicated evidence of enhanced cyclicity when Group A cows were compared with those in Group B. However, there were no significant differences between the two groups for interval to first observed estrus, interval to first serive, first serive pregnancy rate, services per pregnancy and days open. Furthermore, no difference in the incidence of follicular or luteal cysts, incidence of repeat breeders or number of reproductive culls was observed. From observations in this study, the GnRH and PGF₂α treatment scheudule might not be economically beneficial in lactating dairy cows as long as reproductive tract abnormalities are promptly diagnosed and subsequently treated by the attending practitioner.     

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.     

Effects of an analog of PGF2(alpha) (ONO-1052) on cows with luteinized ovarian cysts following treatment with GnRH analog (fertirelin acetate)

Effects of PGF2(alpha) analog (ONO-1052) on cows with luteinized cysts following treatment with GnRH analog (fertirelin) in inducing estrus and shortening an interval from treatment to conception were investigated in a field trial. Seventy-five cows with follicular cysts diagnosed by rectal palpation were treated intramuscularly (i.m.) with 400 mug fertirelin (an analog of GnRH). Luteinization of follicular cysts were tentatively judged by rectal palpation. Forty-one cows were considered to have luteinized cysts and were either treated i.m. with ONO-1052 (28 cows) or left untreated as controls (13 cows). Thirty-four other cows were considered to have not responded to fertirelin and retreated with 10,000 MU hCG (19 cows) or fertirelin (five cows). This tentative judgment of luteinization of the cysts by rectal palpation after treatment was later confirmed by determining serum progesterone concentration before and 10 to 14 days after treatment. Only in the cows with luteinized cysts which were confirmed by serum progesterone analysis; increased from a pretreatment level below 1.0 ng/ml to a value of 1.0 ng/ml or higher 10 to 14 days after treatment, effects of ONO-1052 combined with fertirelin were investigated. Of the 18 cows with luteinized cysts thus confirmed following fertirelin injection and treated with ONO-1052, 15 (83.3 %) cows came into estrus within 26 +/- 14 (mean +/- SD) days after the first treatment; eight cows within three to five days and four cows within 22 to 28 days, and 14 cows (77.8 %) conceived within 100 days (42 +/- 26 days). On the other hand, the five control cows with luteinized cysts that were not treated with ONO-1052 required significantly longer to exhibit normal estrus (54 +/- 13 days; P<0.05) and to conceive (54 +/- 13 days). These results indicate that ONO-1052 combined with fertirelin may be useful to shorten the interval from treatment to normal estrus and to conception in cows with follicular cysts.     

Comparison of the effect of estradiol benzoate plus progesterone and GnRH on the follicular wave emergence and subsequent follicular development in CIDR-treated, lactating dairy cows with follicular cysts

This study examined the effect of estradiol benzoate (EB) plus progesterone (P4) as compared with GnRH on follicular wave emergence and follicular development, and synchrony of ovulation and pregnancy rates following a second injection of GnRH in a controlled internal drug release (CIDR)-based timed AI (TAI) protocol in lactating dairy cows with follicular cysts. Lactating dairy cows diagnosed with follicular cysts received a CIDR device, with an injection of 2mg EB plus 50mg P4 (EB+P4 group) or with an injection of 100 microg GnRH (GnRH group) at the beginning of the experiment (day 0). Thereafter, all received PGF(2alpha) at the time of CIDR removal on day 7, GnRH on day 9, and TAI 16 h later. Follicular wave emergence occurred within 7 days in 12/15 EB plus P4-treated and 14/15 GnRH-treated cows (P>0.05). The interval to wave emergence was longer in the EB+P4 group (4.8+/-0.4 days) than in the GnRH group (2.0+/-0.2 days). The mean diameters of preovulatory follicles and the proportion of cows with preovulatory follicles greater than 12 mm on day 9 did not differ between groups (P>0.05). The proportion of cows with synchronized ovulations by 40 h after the GnRH injection on day 11 and pregnancy rates to TAI did not differ between the EB+P4 (13/15 and 36.7%) and the GnRH (14/15 and 53.3%) groups, respectively. Results suggest that a single treatment with EB plus P4 as compared with GnRH simultaneously with CIDR insertion in lactating dairy cows with follicular cysts will result in relatively asynchronous emergence of a new follicular wave, but subsequently similar sizes of preovulatory follicles and synchronous ovulation, resulting in similar pregnancy rates to TAI.