Synchronising oestrus with oestradiol benzoate after using a two-dose prostaglandin treatment to synchronise luteolysis in dairy heifers

OBJECTIVE: To compare the reproductive performance and pattern of onset of oestrus in dairy heifers in which oestrous cycles were synchronised with two doses of prostaglandin (PG) F2alpha and oestrus was synchronised with oestradiol benzoate (ODB). PROCEDURE: Dairy heifers in two herds (herd A, n = 192; herd B, n = 267) were treated with two doses of an analogue of PGF2alpha (cloprostenol, 375 microg, IM) 12 days apart. Heifers not detected in oestrus 48 h after the last dose of PGF2alpha were either left untreated (No ODB, n = 147) or treated with ODB (0.75 mg IM, n = 126). Onset of oestrus was monitored at 0, 24, 48, 80, 96 and 120 h after the last dose of PGF2alpha Heifers were inseminated on detection of oestrus. RESULTS: After the last dose of PGF2alpha, oestrous detection rates at 80 h (43.5 vs 72.6%, P < 0.001), 96 h (74.1 vs 84.9%, P =0.025) and 120 h (78.2 vs 86.3%, P = 0.082) were less in the No ODB compared to the ODB heifers, respectively. Conception rates (percentage pregnant that were inseminated) were greater in the No ODB compared to the ODB heifers (64.3% vs 47.6%, respectively; P = 0.006), while pregnancy rates (percentage pregnant that were treated) were also greater in the No ODB compared to the ODB heifers, but differences were not significant (50.3% vs 41.1%, respectively; P = 0.068). CONCLUSION: Administration of ODB to heifers not in oestrus 48 h after a two-dose PGF2alpha treatment increases the percentage of heifers detected in oestrus by 80 h, 96 h and 120 h after treatment, by an estimated 29%, 11% and 8%, respectively. However, administration of ODB decreases conception rates by an estimated 17%, and may decrease pregnancy rates (estimated 9% difference). Results are consistent with the hypothesis that ODB can increase submission rates but reduce conception rates following a two dose treatment with PGF2alpha.     

Effectiveness of a novel prostaglandin analog in postpartum mares and mares with early season anestrus

Contents Fifty thoroughbred mares, located at Central-Southern Brazil near Sao Paulo (13 anestrous maiden, 18 anestrous barren mares and 19 postpartum mares) were treated with one to three injections, 48 hrs apart, of 2 mg of the novel PGF analog K 11941*, during the early part of the breeding season. Early season anestrus in these problem mares was compounded by the adverse effects of a drought. Plasma progesterone determinations revealed unexpectedly elevated plasma progesterone levels in 61.5% of the maiden mares, in 77.8% of the barren mares and in 68.4% of the postpartum mares (20 to 25 days postpartum). Treatments with K 11941 initiated luteolysis, heat and ovulations in 91.4, 45.5 an 63.3% of these mares, respectively, but initiated also heat and ovulation in 13% and 33% of mares with baseline progesterone levels. Treatments initiated covert and overt cyclic functions in 17 and 12 of those animals which did not conceive immediately. Fifteen animals with recurrent anestrus received further treatment with K 11941. Ten mares cycling silently were force bred when palpations indicated ovulatory follicles, and 7 conceived. Of the 50 mares treated, 38 became pregnant (76%: maiden mares: 76.9%; barren mares: 77.8% and postpartum mares: 73.7%) with breeding indices of 1.9, 1.36 und 1.57, respectively; and 37 mares gave birth to a live foal (74%), At the same stud, 138 control mares had a foaling rate of 82.6%.     

Induction of luteolysis and oestrus in mares with a synthetic prostaglandin analogue (ICI 81008)

Extract

Cyclical regression of the corpus luteum is induced by an uterine luteolysin which has been shown in several species to be prostaglandin F_2a (PGF_2a , although it is not yet known how it achieves its effect.     

The effect of systemic administration of cloprostenol on ovulation in mares treated with a prostaglandin synthetase inhibitor

Prostaglandins (PGs) are essential to trigger the cascade of events that degrade the extracellular matrix of follicles leading to follicular rupture and ovulation. In mares, systemic administration of flunixin meglumine (FM), a PG synthetase inhibitor, blocks ovulation by inducing luteinized unruptured follicles (LUF). In the rat, the administration of PGF(2α) (PGF) and PGE restored ovulation in indomethacin treated animals. The mares were treated with FM 0, 12, 24 and 36 h after human chorionic gonadotrophin (hCG) administration to induce experimentally LUF (n = 15) or were left untreated (controls, n = 5). In addition, 250 μg of cloprostenol were administered intravenously to the mares 33, 35 and 36 h (CLO 33, n = 5) or 48, 49 and 50 h (CLO 48, n = 5) after hCG. One group was treated with FM but not with cloprostenol (FM-control, n = 5). The ovulation rate, follicular diameter and progesterone concentration were compared amongst groups. The ovulation rate at 48 h was higher (p < 0.05) in the controls (100%) than in the FM-control (0%), CLO 33 (0%) or CLO 48 (20%) mares. All but one FM treated mares developed LUF by 48 h after hCG administration. Two LUF collapsed between 48 and 60 h and 72 and 84 h in one mare from FM-control and from the CLO 33 group each, respectively. Progesterone concentration was significantly higher (p < 0.05) in the control mares than in any of the FM treated mares 5, 9 and 13 days after hCG. In conclusion, FM administered during the periovulatory period blocked ovulation in the mares. In contrast, the administration of cloprostenol, a PGF analogue, in the previously FM treated mares failed to restore ovulation.     

Evaluation of clinical and luteolytic effects of a novel prostaglandin analogue in normal and problem mares

Alfaprostal (K 11941), a novel prostaglandin F2 alpha analogue was clinically tested in 24 dioestrous mares, 40 anoestrous mares and 31 postpartum mares, all being given 2 or 3 mg intramuscularly. Blood samples were taken for the determination of plasma progesterone levels and the objective confirmation of luteolytic drug effects. Two hundred and thirty-six mares at the same location were used for comparisons of the rates of pregnancy, early embryonic loss and foaling. Alfaprostol was found to act as a potent luteolytic agent with good oestrus induction, follicular development and normal conception (72.6%) and foaling rate (65.3%). No side effects were observed. Treatments commenced early in the season (October), when only 35% of the anoestrous mares showed elevated progesterone levels, but in 80% of anoestrous mares with baseline progesterone levels, alfaprostol initiated heat and ovulation followed by normal fertility.     

Melengestrol acetate as a tool for inducing early ovulation in transitional mares

The efficacy of melengestrol acetate (MGA) to shorten the vernal transition of mares by synchronising and accelerating the first ovulation of the year after 60 days of phototherapy was determined by ultrasonographic monitoring. Sixteen mares in late transition were fed two doses of MGA (150 mg/mare/day and 100 mg/mare/day, respectively) for 10 days. A luteolytic dose of prostaglandin was administered to each mare one day after the end of MGA treatment. The presence and duration of oestrus, follicular growth, uterine oedema and presence of ovulation were monitored by ultrasonography and the cervical tone was evaluated by rectal palpation. Ovulation was detected in 87.5% of the mares treated with 150 mg MGA/mare/day for 10 days, and in 62.5% of the mares receiving 100 mg MGA/mare/day for 10 days. This was statistically different (P = 0.03) from the untreated control mares having an ovulation rate of 20%. Mares that received 150 mg MGA/day for 10 days had a mean treatment to ovulation interval of 13.1 +/- 5.97 days after the end of treatment, while mares that received 100 mg MGA/day for 10 days had a mean of 25.6 +/- 10.50 days (P = 0.01) to ovulation. These results suggest that MGA can be used for synchronising and hastening the first ovulation of the year in mares.     

Effects of dihydrotestosterone administration with and without estradiol pretreatment on gonadotropin secretion in ovariectomized pony mares

Twenty ovariectomized pony mares were used to determine if dihydrotestosterone propionate (DHTP) administration, with or without estradiol benzoate (EB) pretreatment, would have the same effects on follicle stimulating hormone (FSH) and luteinizing hormone (LH) secretion as testosterone propionate (TP) administration. All mares were given an initial injection of gonadotropin releasing hormone (GnRH) to characterize their LH and FSH response, and then two groups of mares (n = 4/group) were administered EB (22 micrograms/kg of body weight), two groups were administered vehicle (safflower oil) and a fifth group was administered TP (175 micrograms/kg of body weight) daily for 10 days. Following a second injection of GnRH, one group of EB-treated mares and one group of oil-treated mares were administered DHTP (175 micrograms/kg of body weight) daily for 10 days; the other EB- and oil-treated mares were administered oil and the TP-treated mares were continued on the same dose of TP for 10 days. A final injection of GnRH was then given. Treatment with EB increased (P less than .01) concentrations of LH in daily blood samples and increased (P less than .05) the LH response to exogenous GnRH. Administration of TP or DHTP reduced (P less than .05) both daily LH concentrations and the LH response to exogenous GnRH. Concentrations of FSH in daily blood samples were reduced (P less than .05) and the FSH response to exogenous GnRH was increased (P less than .05) by administration of EB alone, DHTP alone or TP.(ABSTRACT TRUNCATED AT 250 WORDS)     

Intrauterine administration of plant oils inhibits luteolysis in the mare

Reasons for performing the study: The maternal recognition of pregnancy (MRP) signal in the mare has not been determined, although oestrogens have been proposed as a potential candidate. Objectives: To determine effects of intrauterine administration of oestrogen and various oils on cyclic luteolysis in the mare. Hypothesis: Intrauterine oestradiol or fatty acids may suppress luteolysis in the cycling mare when administered during late dioestrus. Methods: A single 1 ml dose of slow‐release oestradiol (10 mg/ml) in fractionated coconut oil was infused into the uterine lumen of cycling mares on Days 6, 8, 10, 12 or 14 post ovulation (n = 12 in each group). Four further groups, each of 12 mares, received an intrauterine infusion of either 1 ml of fractionated coconut oil, peanut oil, mineral oil or a slow‐release preparation of oestradiol (10 mg/ml) in mineral oil on Day 10 post ovulation. Serial blood samples were assayed for progesterone concentrations to monitor luteal function. Results: Intrauterine administration of oestradiol in fractionated coconut oil showed peak efficacy at Day 10 when luteolysis was delayed in 11/12 (92%) mares. The ability of the treatment to delay luteolysis was not significantly different when administered on Days 8 (9/12; 75%), 12 (10/12; 83%) or 14 (6/12; 50%) of dioestrus, but declined significantly when given on Day 6 (3/12; 25%). Oestradiol was not needed to initiate luteostasis since fractionated coconut oil alone or peanut oil administered at Day 10 induced the same high rate of luteal persistence (11/12; 92% for both oils). In contrast, mineral oil did not prolong luteal lifespan, either when administered alone (2/12; 17%) or combined with oestradiol (3/12; 25%). Conclusion: These results do not unequivocally rule out a possible involvement of embryonic oestrogens in MRP in the mare but suggest it is unlikely. The results demonstrate that plant oils can postpone luteolysis, suggesting they may modulate synthesis or release of prostaglandins from the mare's endometrium. Potential relevance: Administration of fractionated coconut or peanut oil on Day 10 post ovulation provides an effective and practical method of prolonging luteal function (‘pseudopregnancy’) thereby suppressing unwanted oestrous behaviour. Further studies to elucidate the mechanism by which this is achieved may increase understanding of both luteostasis and MRP signal in the mare.     

Termination of pseudopregnancy by administration of prostaglandin F2alpha and termination of early pregnancy by administration of prostaglandin F2alpha or colchicine or by removal of embryo in mares.

At day 24 of gestation, pregnant mares were allotted to 1 of 5 treatment groups (3 to 5 mares/group): group A--nontreated controls; group B--intraembryonic injection of 4 mg of colchicine on day 24; group C--removal of embryo on day 24; group D--subcutaneous injection of 1.25 mg of prostaglandin F2alpha (PGF2alpha) on day 32; and group E--removal of embryo on day 24 and subcutaneous injection of PGF2alpha on day 32. In all mares treated with colchicine (group B), the fetal bulge was absent within 2 days. The interval from injection of colchicine to onset of estrus was very short (mean, 4 days). These results indicated that treatment with colchicine was lethal to the 24-day embryo, and pseudopregnancy did not occur. Surgical removal of the embryo (group C) resulted in pseudopregnancy characterized by a prolonged interval from treatment to return to estrus (mean, greater than 31 days), prolonged production of progesterone, and prolonged maintenance of tense uterine and cervical tone. The interval from treatment to ovulatory estrus was longer (P less than 0.05) for group C mares than for group B mares. The mean interval from treatment to complete loss of tense tubular uterine tone was not significantly different between group A pregnant controls (28.3 days) and group C pseudopregnant mares (30 days). Treatment of pregnant mares (group D) with a single injection of PGF2alpha on day 32 resulted in loss of pregnancy in 4 of 4 mares within 2 to 5 days, and in all group D mares a large decrease in progesterone concentration occurred on day 33, 34, or 35. Although subsequent reproductive activity was variable, all group D mares rapidly lost the tense uterine and cervical tone characteristic of early pregnancy. These results indicated that a single subcutaneous injection of 1.25 mg of PGF2alpha caused loss of pregnancy, and pseudopregnancy did not occur. Treatment of group E mares, which had been made pseudopregnant by removal of embryo, with 1.25 mg of PGF2alpha resulted in termination of pseudopregnancy in 5 of 5 mares. All group E mares returned to estrus within 2 to 5 days after treatment, and progesterone concentration decreased (P less than 0.05) within 2 days after treatment. There was no significant difference in loss of tense tubular uterine or cervical tone between pregnant (group D) and pseudopregnant (group E) mares after PGF2alpha treatment.     

Hematological adverse effects and pharmacokinetics of ribavirin in pigs following intramuscular administration

Ribavirin (RBV) is a synthetic guanosine analog that is used as a drug against various viral diseases in humans. The in vitro antiviral effects of ribavirin against porcine viruses were demonstrated in several studies. The purposes of this study were to evaluate the adverse effects and pharmacokinetics of ribavirin following its intramuscular (IM) injection in pigs. Ribavirin was formulated as a double‐oil emulsion (RBV‐DOE) and gel (RBV‐Gel), which were injected into the pigs as single‐dose IM injections. After injection of RBV, all of the pigs were monitored. The collected serum and whole blood samples were analyzed by liquid chromatography–tandem mass spectrometry and complete blood count analysis, respectively. All of the ribavirin‐treated pigs showed significant decreases in body weight compared to the control groups. Severe clinical signs including dyspnea, anorexia, weakness, and depression were present in ribavirin‐treated pigs until 5 days postinjection (dpi). The ribavirin‐treated groups showed significant decrease in the number of red blood cells and hemoglobin concentration until 8 dpi. The mean half‐life of the RBV‐DOE and RBV‐Gel was 27.949 ± 2.783 h and 37.374 ± 3.502 h, respectively. The mean peak serum concentration (C_max) and area under the serum concentration–time curve from time zero to infinity (AUC_inf) of RBV‐DOE were 8340.000 ± 2562.577 ng/mL and 16 0095.430 ± 61 253.400 h·ng/mL, respectively. The C_max and AUC_inf of RBV‐Gel were 15 300.000 ± 3764.306 ng/mL and 207526.260 ± 63656.390 h·ng/mL, respectively. The results of this study provided the index of side effect and pharmacokinetics of ribavirin in pigs, which should be considered before clinical application.     

Effect of administration of prostaglandin F2 alpha on embryo recovery from the uterus on day 5 after ovulation in mares

Ten mares were used to investigate the effect of administration of prostaglandin F2 alpha on uterine tubal motility, as reflected by embryo recovery from the uterus 5 days after ovulation (day 0). Mares were assigned to 3 groups: group A, uterine flush for embryo recovery on day 7; group B, uterine flush for embryo recovery on day 5; and group C, uterine flush for embryo recovery on day 5, after treatment with prostaglandin F2 alpha (10 mg, IM) on day 3. Each mare was assigned to each group once. Embryo recovery rates for the 3 groups were: A, 6 of 10; B, 2 of 8; and C, 0 of 10. The embryo recovery rate for group C was significantly lower (P less than 0.01) than that for group A. Embryo recovery rate for group B was not significantly different from group A or group C. Administration of prostaglandin on day 3 did not increase embryo recovery rate from the uterus on day 5. Additionally, the 25% embryo recovery rate (2 of 8) for group B mares suggested an earlier time for entry of the embryo into the uterus than has previously been reported.     

Effects of fluprostenol administration in mares during late pregnancy

The effectiveness of the prostaglandin F analogue fluprostenol in inducing labour in the mare was examined by giving sequential injections over the last 50 days of gestation. The behavioural and endocrine changes elicited by the drug in pregnant and non-pregnant animals and in foals were also studied. Fluprostenol (250 or 500 micrograms intramuscularly) failed to induce labour before 320 days gestation; thereafter its effect was capricious. Twelve mares foaled 1 to 36 h after the last test; eight delivered normal, viable, apparently 'term' foals and four produced stillborn/premature animals. Eight of the deliveries (five term and three pre-term foals) could be ascribed to the action of fluprostenol because they occurred 1 to 6 h after its administration, at a time when spontaneous foaling would have been unlikely. The other four mares foaled between 12 and 36 h after the fluprostenol injection and it is therefore doubtful whether there was a causal relationship between the two events. In the mares which delivered viable foals the pre-partum milk samples were characteristic of full term samples with respect to calcium, sodium and potassium. Those which delivered premature/stillborn foals had low calcium and a high sodium/potassium ratio in the pre-partum milk. Behavioural changes (sweating, increased respiration, defaecation etc), which varied in intensity between tests and individuals, were seen in all three groups of animals following the administration of fluprostenol. These changes were accompanied by rises in plasma cortisol and adrenocorticotrophic hormone concentration during the 2 h sampling period, suggesting a centrally mediated response to the drug.(ABSTRACT TRUNCATED AT 250 WORDS)     

Exogenous eFSH, follicle coasting, and hCG as a novel superovulation regimen in mares

The objective of this study was to evaluate various equine follicle-stimulating hormone (eFSH) treatment protocols and the effect of “follicle coasting” on ovulation and embryo recovery rates in mares. Cycling mares (n = 40) were randomly assigned to one of four groups 7 days after ovulation: (1) 12.5 mg eFSH twice daily until follicles were 35 mm or larger; (2) 12.5 mg eFSH twice daily until follicles were 32 mm or larger; (3) 12.5 mg eFSH twice daily for 3.5 days followed by 12.5 mg eFSH enriched with luteinizing hormone (LH) twice daily until follicles were 35 mm or larger; and (4) 25 mg eFSH once daily until follicles were 32 mm or larger. Mares in groups 1 and 3 were injected with human chorionic gonadotropin (hCG) (2500 IU intravenously) at the end of eFSH treatment, whereas mares in groups 2 and 4 were given hCG approximately 42 and 54 hours, respectively, after the last eFSH treatment (“follicle coasting”). Nonsurgical embryo collection was performed 6.5 to 7.5 days after ovulation. Each mare experienced a nontreated estrous cycle before being reassigned to a second treatment. Ovulation rates for mares in treatment groups 1 to 4 were 3.3 ± 0.4, 4.1 ± 0.4, 3.5 ± 0.4, and 2.8 ± 0.4 (mean ± SEM; P < .05), respectively. One or more embryos were recovered from more than 80% of mares in each treatment group, and embryo recovery rate per flush was similar among treatment groups (1.9 ± 0.3, 2.6 ± 0.3, 1.9 ± 0.3 and 1.9 ± 0.3, respectively; P > .05). The overall embryo recovery rate was 2.1 ± 1.5 embryos per flush. In summary, ovulation rate was higher for mares treated with eFSH (3.4 ± 0.4) compared with non-treated controls (1.1 ± 0.2). Ovulation rate in mares in which hCG was delayed (follicle coasting) was higher (P < .05) when treatments were given twice per day versus once per day. Administration of equine luteinizing hormone (eLH) in conjunction with eFSH did not have an advantage over mares treated only with eFSH.     

Evaluation of luteolysis and estrous synchronization by a prostaglandin analog (Luprostiol) in Brahman cows and heifers

A trial was conducted to evaluate the ability of a prostaglandin analog, Luprostiol (LP), to synchronize estrus in Brahman cows and heifers. Animals were injected with either 0, 3.75, 7.5, 15 or 30 mg LP or 500 micrograms cloprostenol (CLP) on d 8 or 9 after estrus (d 0). All concentrations of LP (greater than 0 mg) and CLP caused luteolysis in cows and heifers, as indicated by a decline (P less than .01) in serum progesterone concentration after injection. Animals receiving 0 or 3.75 mg LP had a longer (P less than .04) interval to estrus after injection than did animals in other treatment groups. The proportion of animals exhibiting estrus by 120 h after injection was influenced by dose of LP (P less than .0001; 0, 3.75 mg less than 7.5, 15 and 30 mg and CLP) but not by age. Cows had a lower (P less than .01) progesterone concentration than heifers on d 10, 11 and 12 after LP-induced estrus. Progesterone concentration was lowest (P less than .01) on d 10, 11 and 12 after LP-induced estrus in cows given 15 mg LP or CLP. First-service conception rate was similar between cows and heifers, but it was lower (P less than .01) in animals given 15 or 30 mg LP. Both estrogen and LH concentrations were decreased (P less than .01) at the time of estrus by the 15 and 30 mg of LP. Luprostiol can cause luteolysis and estrous synchrony in Brahman cattle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Oxytocin modulates the pulsatile secretion of prostaglandin F2alpha in initiated luteolysis in cattle

Subluteolytic doses of prostaglandin F2alpha analogue (oestrophan) given i.m. and oxytocin (OT) antagonist (CAP) and noradrenaline (NA) infused into the abdominal aorta were used to test the importance of luteal OT in pulsatile secretion of prostaglandin F2alpha (PGF) during luteolysis in heifers (n = 17). In experiment 1, heifers were pre-infused for 30 minutes with saline on either day 17 of the oestrous cycle (group 1; n = 4) or on day 18 of the oestrous cycle (group 2; n = 3), and with CAP (8 mg per animal) on day 17 of the oestrous cycle (group 3; n = 4). Next, heifers were injected with oestrophan (30 microg per animal). Injection of oestrophan in Group 3 increased OT concentrations (P < 0.001) to values similar to those observed during spontaneous luteolysis (50 to 70 pg ml(-1)). PGFM concentrations in this group also increased (P < 0.001), but were lower (P < 0.05) than the values in groups 1 and 2, CAP given prior to oestrophan decreased both PGFM elevation (P < 0.06) and its area under the curve (P < 0.01), compared to the saline pretreated heifers. In experiment 2 NA (4 mg) was infused twice for 30 minutes at five hour intervals to release OT on day 17 of the oestrous cycle (n = 6). However, during hormone analysis it appeared that three of six heifers had elevated PGFM concentrations (group 1) and three others did not (group 2). NA caused the correlated increase of progesterone and OT secretion (r = 0.68; P < 0.05) in both groups but it only influenced PGF secretion in group 1 only (P < 0.05). We postulate that OT can amplify and modulate the course of induced luteolysis as a regulator of the amplitude of pulsatile PGF secretion. PGF analogue stimulates secretion of endogenous PGF from the uterus in cattle and this may be an important component of the luteolytic response to exogenous PGF.