Effect of oestradiol benzoate given after prostaglandin at two stages of follicle wave development on oestrus synchronisation,the LH surge and ovulation in heifers

Oestrus synchronization following prostaglandin-induced luteolysis is variable and dependent on follicle wave status in cattle. Oestradiol benzoate (ODB) has been used following prostaglandin to reduce the interval to oestrus and ovulation, but the effect of follicle wave status at the time of ODB administration is not clear. The aim of this study was to characterize the endocrine and follicular responses following ODB after luteolysis at different stages of the follicle wave. Prostaglandin was administered at either emergence or dominance of the second follicle wave. Twenty-four hours later animals received either 0.5mg ODB in oil or a control oil injection. Follicular development was monitored daily by ultrasonography, oestrous behavior was determined and blood samples were collected. In animals treated with ODB at emergence, there was a reduction (P<0.05) in the maximum diameter of the ovulatory follicle (11.7+/-1.2 mm versus 13.1+/-0.1 mm) and in the interval from prostaglandin to oestrus (52.0+/-2.3 h versus 88.0+/-9.6h), to the LH surge (53.3+/-3.5 h versus 89.1+/-6.5 h) and to ovulation (96+/-0.0 h versus 129.6+/-9.6h), compared with controls. In animals treated with ODB at dominance, there was a reduction (P<0.05) in the interval from prostaglandin to the LH surge (54.0+/-3.1 h versus 70.9+/-4.8 h), but not in the interval from prostaglandin to oestrus (53.3+/-2.7 h versus 65.7+/-4.5 h; P=0.11), to ovulation (96.0+/-0.0 h versus 110.4+/-4.8 h; P=0.12) or the maximum diameter of the ovulatory follicle (12.7+/-0.3 mm versus 13.6+/-0.4 mm; P=0.12), compared with controls. Treatment did not affect (P>0.05) the length of the subsequent oestrous cycle or corpus luteum size. In conclusion, the use of ODB advanced, but did not alter the temporal relationships among oestrus, the LH surge and ovulation, regardless of stage of follicle development at treatment.     

The effect of dose and route of oestradiol benzoate administration on plasma concentrations of oestradiol and FSH in long-term ovariectomised heifers

Oestradiol (E(2)) suppresses FSH and affects follicle wave dynamics in cattle. However, neither the optimum dose of ODB required to suppress FSH nor the effect of route of ODB administration on blood concentrations of E(2) are known; hence, the aim of this experiment was to answer these questions. Ovariectomised heifers received Progesterone Releasing Intravaginal Device (PRID) for 7 days, and 4 days later heifers received one of eight ODB treatments at second PRID insertion as follows; (1) 0.0 mg (Control; n=3), (2) 0.5 mg (n=4), (3) 1.0 mg (n=4), (4) 2.5 mg (n=6), (5) 5.0 mg (n=4), (6) 10. 0 mg (n=4), (7) 5.0 mg (n=4), and (8) 10.0 mg (n=5). For treatments 2-6 inclusive, ODB was administered intramuscularly in oil, while for treatments 7 and 8, the ODB in powder form was administered topically in the vagina by gelatine capsule attached to the PRID. Blood samples were collected every 6 h for the first 48 h, every 12 h for the next 48 h, and twice daily for a further 6 days. The interval from ODB administration to peak E(2) concentration was similar (P0.05) for treatments 2-6 where ODB was administered intramuscularly (mean 13.4+/-1.24 h), and was longer (P<0.05) for the intravaginal capsule treatments (mean 25.5+/-2.84 h). Plasma concentrations of E(2) increased with increasing intramuscular dose of ODB injected, (plasma E(2)=-0.237+16.109 (dose)-0.74 (dose)(2), R(2)=0.75; P<0.05). Peak plasma concentrations of E(2) following the 5- and 10-mg capsules were similar to each other and to those following the 0.5-mg injection (P0.05), but were lower than concentrations obtained following injection of 1.0-5.0 mg (P<0.05). Across all treatments, both the maximum percentage decline in FSH and the interval to FSH nadir were related to the peak plasma concentrations of E(2) (maximum % decline in FSH=11.17+1.564 (peak E(2))-0.009 (peak E(2))(2), R(2)=0.75; P<0.01), (hours to FSH nadir=10.628+1.486(hours to peak E(2))-0.0282(hours to peak E(2))(2), R(2)=0.22; P<0.05). Concentrations of FSH increased as E(2) declined from its peak value, irrespective of maximum value achieved. It was concluded that the intramuscular administration of ODB in oil to ovariectomised heifers given a PRID results in higher plasma concentrations of E(2) and causes a greater reduction in FSH than administration topically by intravaginal gelatine capsule. E(2) transiently suppresses FSH in ovariectomised heifers, and the magnitude of the suppression is dose-dependent; however FSH concentrations begin to increase 1-2 days after ODB administration while concentrations of E(2) were declining but still high.     

Ovarian responses to progesterone and oestradiol benzoate administered intravaginally during dioestrus in cattle

This study examined the effects of administering progesterone and oestradiol benzoate (ODB) during mid-dioestrus, on ovarian follicular dynamics in cattle. Twelve cycling cows were used in a 4 x 4 latin square design, with the 4 treatments being initiated on Day 13 of the cycle (oestrus = Day 0) and comprising intravaginal insertion for 5 days of: (i) a progesterone releasing device (CIDR; 'P4'); (ii) a CIDR device with a gelatin capsule containing 10 mg ODB and 1 g lactose (CIDIROL; 'P4/ODB') attached; (iii) a placebo CIDR device with the 10 mg ODB capsule (ODB); and, (iv) a placebo CIDR device alone (CTRL). The ovaries of each cow were examined daily by transrectal ultrasonography from Day 7 of the cycle until subsequent ovulation. Blood samples were collected daily from Day 11, and at intervals of 2-4 h during the 24 h period either side of treatment initiation. The second dominant follicle (DF2) emerged on Day 10.7 +/- 0.2 (mean +/- SEM), and was 8.5 +/- 0.2 mm in diameter by Day 13. The DF2 developed through to ovulation (2-wave cycles) in half of the animals in the CTRL group; while in the other half of cases, the ovulatory follicle originated from the third follicle wave that emerged on Day 17.2 +/- 0.4. Administration of a CIDR device alone (P4 group) did not alter the 1:1 ratio of 2 and 3-wave cycles, but the third dominant follicle (DF3) in those cows with 3-wave cycles emerged earlier on Day 15.6 +/- 0.2. In contrast, the DF2 of every animal in the ODB and P4/ODB groups became atretic and was replaced by a DF3 which emerged 4.0 +/- 0.3 days later. The effects of ODB on luteal function were limited to an earlier decline in plasma progesterone concentrations from 2 to 4 days after device insertion and a reduction in diameter of the corpus luteum when administered concurrently with progesterone. Intravaginal administration of 10 mg ODB on Day 13 of the oestrous cycle, with or without progesterone, was effective in promoting follicle wave turnover. In the absence of ODB, progesterone administration alone (P4 group) did not alter the ratio of animals with 2 or 3-wave cycles from that observed in animals in the CTRL group, but did advance the timing of subsequent follicle wave emergence in those animals with 3-wave cycles.     

Effect of early luteolysis in progesterone-based timed AI protocols in Bos indicus, Bos indicus x Bos taurus, and Bos taurus heifers

The objective of this study was to evaluate the effects of treatment with an intravaginal progesterone-releasing device (CIDR) and estradiol benzoate (EB) on follicular dynamics in Bos indicus (n=23), Bos taurus (n=25), and cross-bred (n=23) heifers. To assess the influence of reduced serum progesterone concentrations during 8 days of treatment with a progesterone-releasing device on follicular dynamics, half of the heifers received PGF at CIDR insertion (Day 0; 3 x 2 factorial design). Mean (+/-S.E.M.) serum progesterone concentrations during CIDR treatment varied (P<0.05) among genetic groups: B. indicus (5.4+/-0.1 ng/mL), B. taurus (3.3+/-0.0 ng/mL), and cross-bred (4.3+/-0.1 ng/mL). Maximum diameter of the dominant follicle (DF) was smaller (P<0.01) in B. indicus heifers (9.5+/-0.5 mm) than in cross-bred (12.3+/-0.4 mm) or B. taurus heifers (11.6+/-0.5 mm). B. indicus experienced lower (P<0.01) ovulation rate (39.1%) than did B. taurus (72.7%) and cross-bred (84.0%). Heifers treated with PGF on Day 0 had lower (P<0.05) serum progesterone concentrations during progesterone treatment. The PGF treatment on Day 0 increased (P<0.01) the diameter of the DF (11.9+/-0.4 mm vs. 10.5+/-0.4 mm). Moreover, greater (P=0.02) ovulation rates (78.8 vs. 54.0%) occurred in heifers treated with PGF on Day 0. In summary, B. indicus heifers had greater serum progesterone concentrations, smaller DF diameter, and a lower ovulation rate compared to B. taurus heifers. Prostaglandin treatment on the day of CIDR insertion reduced serum progesterone during treatment, and resulted in increased maximum DF diameter and ovulation rate.     

Ovarian follicular development in Holstein cows following synchronisation of oestrus with oestradiol benzoate and an intravaginal progesterone releasing insert for 5-9 days and duration of the oestrous cycle and concentrations of progesterone following ovulation

The aim of this study was to determine if the duration of treatment with an intravaginal progesterone releasing insert (IVP(4)) after treatment with oestradiol benzoate (ODB) at the time of insertion and 24 h after removal would affect selected variables including: size of ovarian follicles at the time of removal of inserts, diameter of ovulatory follicles, plasma concentrations of progesterone following ovulation, and duration of the following oestrous cycle. Characteristics of oestrus at a synchronised and spontaneous oestrus were also monitored. Non-lactating Holstein cows were synchronised with an IVP(4) for 5 (n = 10), 7 (n = 10), 8 (n = 9) or 9 (n = 9) days together with injections of ODB at device insertion (2 mg) and 24 h after removal (1 mg). Ultrasonography showed no significant effect of treatment on the day of emergence of preovulatory follicles relative to the day of removal of inserts (overall mean = -4.22 +/- 0.58; P = 0.15) for cows that ovulated within 120 h insert removal (n = 36). Treatment with ODB and an IVP(4) for 5 days reduced the diameter of preovulatory follicles at the time of removal of inserts and for the following 2 days compared to cows treated for 7-9 days (mean difference 2.56 +/- 1.15 mm; P = 0.033) but did not reduce the diameter of the ovulatory follicle (P = 0.21). Day of emergence relative to removal of inserts was associated with the diameter of the ovulatory follicle (R2 = 0.69; P < 0.001). Concentrations of progesterone and the diameter of the corpus luteum following ovulation were not affected by treatment (P > 0.20), but were affected by the diameter of the ovulatory follicle (P < 0.01). Diameter of the ovulatory follicle did not affect interoestrous and interovulatory intervals (P > 0.40). We conclude that treatment with an IVP(4) for 5 compared to 7-9 days with ODB administered at device insertion, and 24 h after removal reduced the diameter of preovulatory follicles at the time of removal of the insert but did not reduce the diameter of the ovulatory follicle or concentrations of progesterone in plasma. Emergence of preovulatory follicles closer to the time of removal of inserts reduced the diameter of the ovulatory follicle when oestrus was induced with ODB. Ovulation of smaller follicles reduced concentrations of progesterone in plasma following ovulation but did not affect oestrous cycle duration.     

Effects of estradiol cypionate (ECP) on ovarian follicular dynamics,synchrony of ovulation,and fertility in CIDR-based,fixed-time AI programs in beef heifers

Estradiol cypionate (ECP) was used in beef heifers receiving a controlled internal drug release (CIDR; insertion = Day 0) device for fixed-time AI (FTAI) in four experiments. In Experiment 1, heifers (n = 24) received 1mg ECP or 1mg ECP plus 50mg commercial progesterone (CP) preparation i.m. on Day 0. Eight or 9 days later, CIDR were removed, PGF was administered and heifers were allocated to receive 0.5mg ECP i.m. concurrently (ECP0) or 24h later (ECP24). There was no effect of treatment (P = 0.6) on mean (+/-S.E.M.) day of follicular wave emergence (3.9+/-0.4 days). Interval from CIDR removal to ovulation was affected (P<0.05) only by duration of CIDR treatment (88.3+/-3.8h versus 76.4+/-4.1h; 8 days versus 9 days, respectively). In Experiment 2, 58 heifers received 100mg progesterone and either 5mg estradiol-17beta or 1mg ECP i.m. (E-17beta and ECP groups, respectively) on Day 0. Seven (E-17beta group) or 9 days (ECP group) later, CIDR were removed, PGF was administered and heifers received ECP (as in Experiment 1) or 1mg EB 24h after CIDR removal, with FTAI 58-60h after CIDR removal. Follicular wave emergence was later (P<0.02) and more variable (P<0.002) in heifers given ECP than in those given E-17beta (4.1+/-0.4 days versus 3.3+/-0.1 days), but pregnancy rate was unaffected (overall, 69%; P = 0.2). In Experiment 3, 30 heifers received a CIDR device and 5mg E-17beta, with or without 100mg progesterone (P) i.m. on Day 0. On Day 7, CIDR were removed and heifers received ECP as described in Experiment 1 or no estradiol (Control). Intervals from CIDR removal to ovulation were shorter (P<0.05) in ECP0 (81.6+/-5.0h) and ECP24 (86.4+/-3.5h) groups than in the Control group (98.4+/-5.6h). In Experiment 4, heifers (n = 300) received a CIDR device, E-17beta, P, and PGF (as in Experiment 3) and after CIDR removal were allocated to three groups (as in Experiment 2), with FTAI 54-56h (ECP0) or 56-58h (ECP24 and EB24) after CIDR removal. Pregnancy rate did not differ among groups (overall, 63.6%, P = 0.96). In summary, although 1mg ECP (with or without progesterone) was less efficacious than 5mg E-17beta plus 100mg progesterone for synchronizing follicular wave emergence, 0.5mg ECP (at CIDR removal or 24h later) induced a synchronous ovulation with an acceptable pregnancy rate to fixed-time AI.     

Effect of treatment with progesterone and oestradiol benzoate on ovarian follicular turnover in postpartum anoestrous cows and cows which have resumed oestrous cycles.

Two experiments were carried out to determine the effect of a low dose of progesterone (P) with and without the addition of an injection of oestradiol benzoate (ODB) on ovarian follicle dynamics, oestradiol production and LH pulsatility in postpartum anoestrous cows, compared with cows which had resumed oestrous cycles (cycling cows). In the first experiment, anoestrous Jersey cows were treated with (AN+P, n=8) or without (AN-3, n=3) a previously used intravaginal progesterone releasing (CIDR) device for 10 days, commencing 3 or 4 days after emergence of a new dominant follicle (DF1) as determined by transrectal ultrasonography. Contemporary cycling cows (CYC+P, n=8) were similarly treated with used CIDR devices and injected with prostaglandin F(2alpha) (PGF) at the time of device insertion. Follicle turnover was monitored by daily ultrasonography and pulsatile release of LH was measured on the ninth day after device insertion. During the period of CIDR device insertion, a second dominant follicle emerged in 4/8 of the CYC+P group and 7/8 of the AN+P group (P=0.14). Maximum diameter of DF1 was greater in cows in the CYC+P compared with the AN+P group (P=0.02), but did not differ between cows in the AN+P and AN-P groups (P>0.1). Frequency of LH pulses was greater in cows in the CYC+P than AN+P group (P=0.06), and in cows in the AN+P than AN-P group (P=0.02).In the second experiment, anoestrous (n=20) and cycling (n=11) Friesian cows were treated with a new CIDR device for 6 days commencing 3 days after emergence of a new dominant follicle (DF1). Cycling cows were also injected with PGF on the day of device insertion. Half of the cows in each group were injected with 2mg ODB on the day of device insertion. Daily ultrasonography was used to monitor follicular dynamics throughout the experimental period. Follicular turnover was increased by ODB in cycling (5/5 versus 1/6; P<0.05), but not anoestrous cows (5/9 versus 4/11). Persistence of DF1 was reduced by ODB treatment in both cycling and anoestrous cows (P<0.001). Maximum diameter of DF1 was influenced by ODB treatment and reproductive status (P<0.05). In anoestrous cows in which a second dominant follicle did not emerge during the period of device insertion, the interval from emergence of DF1 to emergence of a second dominant follicle was significantly delayed by treatment with ODB (P=0.04).In conclusion, P treatment of anoestrous cows increased pulsatile release of LH, but did not induce the development of persistent follicles. Injection of ODB in association with P treatment reduced the persistence of dominant follicles in both cycling and anoestrous cows, but delayed subsequent follicular development in a proportion of anoestrous cows.     

Effect of exogenous progesterone and oestradiol on plasma progesterone concentrations and follicle wave dynamics in anovulatory anoestrous post-partum dairy cattle

The effect of exogenous progesterone (P4) and of oestradiol benzoate (ODB) on plasma progesterone concentration and follicle dynamics was studied in anovulatory anoestrus (AA) post-partum pasture-fed dairy cattle. Cows (n=32) were defined AA based on not detecting a corpus luteum upon transrectal ultrasonography of the ovaries. Cows were randomly assigned to treatment with an intravaginal P4-releasing device containing 1.56 g of P4 (1Q; Cuemate, Pfizer Animal Health, Auckland, NZ; n=11) or with modified devices with double (2Q; n=11) or triple (3Q; n=10) the normal P4 dose for 8 days. Half of each group received 2 mg ODB at device insertion (Day 0) while the other half did not receive ODB at this time. All cows were treated with 1 mg ODB 1 day after intravaginal device removal (Day 9). Ultrasonography occurred daily until either ovulation or Day 15 whichever occurred sooner. Blood samples were drawn on Days 0, 1, 3, 5, 7, 8, 9, 15 and 22 for plasma P4 determination. Increasing P4 dose was associated with an increase in plasma P4 concentration during the time of device insertion (P <0.05). The highest P4 dose was associated with a delay in emergence of, but a shorter interval from emergence to maximum diameter and ovulation of, the subsequent dominant follicle (DF2) compared to the lowest P4 dose. Treatment with ODB resulted in a delay in emergence of DF2 (4.2 (0.4) versus 2.0 (0.4) days (S.E.M.) to emergence for ODB versus no-ODB; P=0.01), a smaller maximum diameter of DF2 (15.2 (0.5) versus 17.9 (0.6)mm (S.E.M.) for ODB versus no-ODB; P <0.01), a shorter interval to maximum DF2 diameter (5.0 (0.3) versus 6.8 (0.3) days (S.E.M.) for ODB versus no-ODB; P=0.03), a shorter interval from DF2 emergence to ovulation (6.3 (0.4) versus 8.5 (0.4) days (S.E.M.) for ODB versus no-ODB; P=0.02) and a tendency for a lower average plasma P4 concentration post-ovulation (i.e. average of Days 15 and 22; 2.5 (0.4) versus 3.4 (0.4) ng/ml plasma P4 for ODB versus no-ODB, respectively; P=0.08). The DF present at device insertion, was still present at device removal in three (9%) cows of which two were treated with 1Q + no-ODB and one with 3Q + ODB. It is concluded that increasing P4 dose and ODB treatment are associated with a delay in subsequent follicle wave emergence and more rapid follicle growth. Oestradiol benzoate treatment also tends to reduce the plasma P4 concentration in the subsequent luteal phase in post-partum, anoestrous dairy cattle.     

Induction of follicular wave emergence for estrus synchronization and artificial insemination in heifers

The objective was to synchronize follicular wave emergence among cattle for synchronization of estrus and ovulation, and to determine pregnancy rate after AI at observed estrus. At random stages of the estrous cycle, a controlled internal drug release device (CIDR-B) was inserted intravaginally (Day 0) in 67 cross-bred beef heifers, and they were randomly allocated to receive either no further treatment (Control; n = 18); 5 mg of estradiol-17beta and 100 mg of progesterone im (E/P; n = 16); 100 microg im of GnRH (GnRH; n = 16); or transvaginal ultrasound-guided follicular ablation of all follicles > or = 5 mm (FA; n = 17). All heifers received a luteolytic dose of PGF (repeated 12 h later), and CIDR-B were removed on Days 9, 8, 6 or 5, in Control, E/P, GnRH or FA groups, respectively, so the dominant follicle of the induced wave was exposed to exogenous progesterone for a similar period of time in each group. Mean (+/- SEM) intervals (and range, in days) from treatment to follicular wave emergence in these groups were 3.5 +/- 0.6 (-2 to 8), 3.4 +/- 0.1 (3 to 4), 1.5 +/- 0.3 (-1 to 4), and 1.0 +/- 0.1 (0 to 2), respectively. Although the interval was longest (P<0.01) in the E/P and Control groups, it was least variable (P<0.01) in the E/P and FA groups. Intervals (and range, in days) from CIDR-B removal (and first PGF treatment) to estrus were 2.3 +/- 0.2 (1.5 to 4.5), 2.2 +/- 0.2 (1.5 to 3.0), 2.1 +/- 0.1,(1.5 to 3.5), and 2.5 +/- 0.1 (2.0 to 3.5), and to ovulation were 3.5 +/- 0.2 (2.5 to 5.5), 3.4 +/- 0.1 (3.0 to 4.5), 3.5 +/- 0.1 (2.5 to 4.5), and 3.8 +/- 0.1 (3.0 to 4.5), for Control, E/P, GnRH and FA groups, respectively (ns). The proportion of heifers displaying estrus was higher in the Control than in the FA group (94% versus 65%, P<0.05) and intermediate in EP and GnRH groups (87% and 75%). Heifers were inseminated approximately 12 h prior to ovulation (based on estrous behavior and ultrasound examinations). Pregnancy rates were 78%, 80%, 69% and 65% for Control, E/P, GnRH and FA groups, respectively (P=0.73). Results support the hypothesis that synchronous follicular wave emergence results in synchronous follicle development and, following progesterone removal, synchronous estrus and ovulation with high pregnancy rates to AI. The synchrony of estrus and ovulation in the E/P, GnRH and FA groups suggest that these treatments, in combination with CIDR-B, could be adapted to fixed-time insemination programs.     

Estradiol benzoate delays new follicular wave emergence in a dose-dependent manner after ablation of the dominant ovarian follicle in cattle

Administration of estradiol benzoate (EB) induces atresia of the dominant follicle (DF) in the ovaries of cattle within 36 h but emergence of a new wave of follicular development is delayed by 3-5 days. The present study investigated the role of EB in determining timing of emergence of a new follicular wave after removing the influence of the DF. At 6.4+/-0.2 days after ovulation in Angus and Angus/Simmental cattle (n=26), aged 4.9+/-0.6 years and weighing 634+/-20 kg, all ovarian follicles > or =5mm in diameter were aspirated with a 17-gauge needle using an ultrasound-guided transvaginal approach (Day 0 or Hour 0) and animals immediately received 0 (0EB), 1 (1EB), 2 (2EB) or 4 (4EB) mg EB i.m./500 kg body weight (n=6 or 7 per treatment). Ovarian structures were monitored by ultrasonography on a daily basis until emergence of a new wave of follicular development. Concentrations of estradiol (E2) were different among all treatments between Hours 24 and 72, increasing (P<0.01) with greater doses of EB administered. Hour of peak follicle-stimulating hormone (FSH) was 29.3+/-4.0, 53.3+/-4.5, 81.1+/-15.5, and 91.4+/-8.2 for the 0EB, 1EB, 2EB, and 4EB treatments, respectively, and emergence of a new wave of follicular development occurred on Days 1.5+/-0.2, 3.3+/-0.3, 4.0+/-0.6 and 4.4+/-0.4, respectively. Timing of peak FSH and emergence of a new wave of follicular development was earliest (P<0.05) in the 0EB treatment, similar (P>0.1) among the 1EB and 2EB treatments, and most delayed (P<0.05) in the 4EB treatment when compared to the 0EB or 1EB treatments. The overall mean interval from peak FSH to emergence of a new wave of follicular development was 15.7+/-3.3 h and was not affected by treatment. Concentrations of E2 at 24 h before new emergence were not different among EB-treated animals (20.2+/-5.5 pg/ml), but lower (P<0.01) in the 0EB treatment (1.6+/-0.2 pg/ml). In a dose-dependent manner, EB delayed the pre-emergence surge in FSH that stimulates new follicular development after the DF has ceased to be functional. The importance of using an 'optimal' dose of EB in hormonal regimens using this agent to strategically regulate follicular development is emphasized by the outcomes of this study.     

Follicle deviation and ovulatory capacity in Bos indicus heifers

The objectives of Experiment 1 were to determine the interval from ovulation to deviation, and diameter of the dominant follicle (DF) and largest subordinate follicle (SF) at deviation in Nelore (Bos indicus) heifers by two methods (observed and calculated). Heifers (n = 12) were examined ultrasonographically every 12 h from ovulation (Day 0) to Day 5. The time of deviation and diameter of the DF and largest SF at deviation did not differ (P>0.05) between observed and calculated methods. Overall, deviation occurred 2.5+/-0.2 d (mean +/- S.E.M.) after ovulation, and diameters for DF and largest SF at deviation were 6.2+/-0.2 and 5.9 +/- 0.2 mm, respectively. Experiment 2 was designed to determine the size at which the DF acquires ovulatory capacity in B. indicus heifers. Twenty-nine heifers were monitored every 24 h by ultrasonography, from ovulation until the DF reached diameters of 7.0-8.4 mm (n=9), 8.5-10.0 mm (n=10), or >10.0 mm (n=10). At that time, heifers were treated with 25 mg of pLH and monitored by ultrasonography every 12 h for 48 h. Ovulation occurred in 3 of 9, 8 of 10, and 9 of 10 heifers, respectively (P<0.05). In summary, there was no significant difference between observed and calculated methods of determining the beginning of follicle deviation. Deviation occurred 2.5 d after ovulation when the DF reached 6.2 mm, and ovulatory capacity was acquired by DF as small as 7.0 mm.     

Effect of treatment with progesterone and oestradiol when starting treatment with an intravaginal progesterone releasing insert on ovarian follicular development and hormonal concentrations in Holstein cows

Ovarian follicular development and concentrations of gonadotrophin and steroid hormones were studied in non-lactating Holstein cows following administration of progesterone (P(4)) or oestradiol benzoate (ODB) at the start of treatment with an intravaginal progesterone releasing insert (IVP(4)) in a 2 by 2 factorial experiment. Cows were treated at random stages of the oestrous cycle with an IVP(4) device (Day 0) and either no other treatment (n=8), 200 mg of P(4) IM (n=9), 2.0 mg of ODB IM (n=8) or both P(4) and ODB (n=9). Seven days later devices were removed and PGF(2alpha) was administered. Twenty-four hours later 1.0mg of ODB was administered IM. Oestrus was detected in 97.1% and ovulation in 64.7% (effect of treatment, P=0.41) of cows within 96 h of removing inserts. In the cows that ovulated, day of emergence of the ovulatory follicle was delayed (P<0.01) and more precise (P<0.05) in cows treated with ODB compared to the cows treated with P(4). Interval from wave emergence to ovulation and the diameter of the ovulatory follicle was less (P<0.05) in cows treated with ODB compared to cows treated with P(4). Combined treatment with P(4) and ODB at the time of starting treatment with an IVP(4) device did not significantly change the pattern of ovarian follicular development compared to treatment with ODB alone. Concentrations of LH and FSH in plasma were less in cows treated with ODB between Days 0 and 4 (P<0.05) while treatment with P(4) increased concentrations of FSH in plasma between Days 0 and 4 (P<0.05). When anovulatory cows were compared to ovulatory cows, diameters of follicles (P<0.001) and growth rate of follicles (P<0.01) were less in anovulatory cows between Days 7 and 9, while concentrations of FSH in plasma were greater (P<0.01), concentrations of LH similar (P>0.90) and concentrations of oestradiol were less (P=0.01) in the anovulatory cows between Days 4 and 10. Our findings support a hypothesis that ovarian follicular development following administration of P(4) or ODB at the start of treatment with an IVP(4) device differs. Anovulatory oestrus may have been associated with reduced maturity and/or later emergence of ovarian follicles.     

Effect of dietary intake on pattern of growth of dominant follicles during the oestrous cycle in beef heifers

Friesian x Hereford heifers (n = 19; mean +/- s.e.m. body weight (BW) = 375 +/- 5 kg) were used in a randomized incomplete block design. Heifers were fed 0.7 (n = 7; L), 1.1 (n = 7; M) or 1.8% (n = 5; G) of BW in dry matter (DM)/day for 10 weeks. Ovaries were examined by ultrasound, for one oestrous cycle, from week 5 of treatment. Maximum diameter of dominant follicles was smaller (P less than 0.05) in L (11.8 +/- 0.1 mm) than in M (13.7 +/- 0.2 mm) or G (13.2 +/- 0.3 mm) heifers. Growth rate (mm/day) of dominant follicles during the oestrous cycle was not affected (P greater than 0.05) by dietary intake. Persistence of dominant follicles was shorter (P less than 0.05) in L (9.8 +/- 0.2 days) than in M (11.9 +/- 0.3 days) or G (12.7 +/- 0.4 days) heifers. Three dominant follicles were identified during the oestrous cycle of 5 of 7 L, 3 of 7 M and 1 of 5 G heifers (P less than 0.10); 2 dominant follicles were identified in the remaining heifers (n = 2 of 7, 4 of 7 and 4 of 5, respectively). Length of the luteal phase and luteal-phase concentrations of progesterone were not affected (P greater than 0.05) by treatment. Low dietary intake reduced the diameter and persistence of dominant follicles during the oestrous cycle of beef heifers and tended to increase the proportion of oestrous cycles with 3 dominant follicles.     

Evaluation of two different oestrus-synchronisation methods with timed artificial insemination and resynchronisation of returns to oestrus in lactating Holstein cows

To examine the outcomes of adding medroxyprogesterone acetate (MAP) to the ovsynch protocol with the traditional ovsynch protocol in both cycling and anoestrus cows, and to evaluate a resynchronisation protocol, 742 cows averaging more than 40 days postpartum were assigned to the following four treatments: (1) ovsynch (OVS): day 0: GnRH; day 7: PGF2alpha; day 9: a similar dose of GnRH; day 10: timed artificial insemination (TAI), approximately 16-20h later; (2) ovsynch+MAP (MAP): the same ovsynch protocol plus an intravaginal insert made of polyurethane sponge impregnated with 300mg of MAP immediately after the first GnRH treatment and on day 7, at the time of the PG treatment, the sponge was removed; (3) resynchronisation (MAP+ODB): 1mg of oestradiol benzoate (ODB) on day 13 after TAI and a new sponge impregnated with MAP was inserted and; on day 20, 1mg of ODB was given and the sponge removed; and (4) no resynchronisation (No MAP): only oestrus detection and AI at any repeat oestrus detected after TAI. Progesterone was measured in milk samples collected on days -17, -10, -3, 13 and 20 (TAI=day 0). Based on milk P4 at days -17 and -10, 27.4% of the cows were still anoestrus. At PG injection, 67.7% of the cycling and 21.3% of the anoestrus cows had elevated P4. Farm, days postpartum and parity variations were detected in both cases. On day 20 after TAI 42.6% of cycling and 8.3% of the anoestrous cows had elevated P4. Pregnancy rates were similar in both pre-breeding treatments (20%), but interactions (P<0.001) were detected between treatment and cycling activity (for anoestrous cows: MAP=34.9%; OVS=11.1%. Average interval from TAI to subsequent AI was 37+/-3 days. Resynchronisation resulted in more (P<0.001) cows in oestrus between days 18 and 25 after TAI. Conception rate in the MAP+ODB treatment was lower (P<0.05) than the No MAP group (22.8% versus 47.4%). It was concluded that the addition of a progestin to the ovsynch protocol resulted in increased pregnancy rates of cows treated during anoestrus. The benefit of including MAP with the ovsynch protocol for cycling cows is equivocal.     

Ovarian responses in Bos indicus heifers treated to synchronise ovulation with intravaginal progesterone releasing devices, oestradiol benzoate, prostaglandin F(2α) and equine chorionic gonadotrophin

The objectives were: (i) improve understanding of the ovarian responses of Bos indicus heifers treated with different ovulation synchronisation protocols, (ii) compare ovarian responses of B. indicus heifers treated with intravaginal progesterone releasing device (IPRD)+oestradiol benzoate (ODB) versus a conventional prostaglandin F(2α) (PGF(2α)) protocol and (iii) investigate whether reducing the amount of progesterone (P(4)) in the IPRD, and treatment with equine chorionic gonadotrophin (eCG) would increase the proportion of heifers with normal ovarian function during the synchronised and return cycles. Two-year-old Brahman (n=30) and Brahman-cross (n=34) heifers were randomly allocated to three IPRD-treatment groups: (i) standard-dose IPRD (Cue-Mate(?) 1.56g P(4); n=17); (ii) half-dose IPRD (Cue-Mate(?) 0.78g P(4); n=15); (iii) half-dose IPRD+300IU eCG at IPRD removal (n=14), and a non-IPRD control group (iv) 2×PGF(2α) (500μg cloprostenol) on Days -16 and -2 (n=18). IPRD-treated heifers received 250μg cloprostenol at IPRD insertion (Day -10) and IPRD removal (Day -2) and 1mg ODB on Days -10 and -1. Ovarian function was evaluated by ultrasonography and plasma P(4) throughout the synchronised and return cycles. The mean diameter of the dominant follicle observed at 54-56h after IPRD removal, was greater for heifers which ovulated than heifers which did not ovulate (P<0.001; 14.5±1.1 vs. 9.3±0.6mm, respectively). The prevalence of IPRD-treated heifers with ovarian dysfunction (persistent CL, failure to re-ovulate, shortened luteal phase) was 39%. This relatively high prevalence of ovarian dysfunction may explain the commonly reported, lower than expected pregnancy rates to FTAI in B. indicus heifers treated to synchronise ovulation.     

Induction of a new follicular wave in holstein heifers synchronized with norgestomet

Treatments with progestin to synchronize the bovine estrous cycle in the absence of the corpus luteum, induces persistence of a dominant follicle and a reduction of fertility at doses commonly utilized. The objective of the present research was to induce a new wave of ovarian follicular development in heifers in which stage of the estrous cycle was synchronized with norgestomet. Holstein heifers (n=30) were used, in which estrus was synchronized using two doses of PGF2alpha i.m. (25 mg each) 11 days apart. Six days after estrus (day 0=day of estrus) heifers received a norgestomet implant (6 mg of norgestomet). On day 12, heifers were injected with 25 mg of PGF2alpha i.m. and assigned to treatments (T1 to T4) as follows: treatment 1, heifers received a second norgestomet implant (T1: N+N, n=6), treatment 2, received 100 microg of GnRH i.m. (T2: N+GnRH, n=6), treatment 3, 200 mg of progesterone i.m. (T3: N+P4, n=6), treatment 4, control treatment with saline solution i.m. (T4: N+SS); in the four treatments (T1 to T4) implants were removed on day 14. For treatment 5, heifers received 100 microg of GnRH i.m. on day 9 and 25 mg of PGF2alpha i.m. (T5: N+GnRH+PGF2alpha) at the time of implant removal (day 16). Ovarian evaluations using ultrasonographic techniques were performed every 48 h from days 3 to 11 and every 24 h from days 11 to 21. Blood samples were collected every 48 h to analyze for progesterone concentration. A new wave of ovarian follicular development was induced in 3/6, 6/6, 3/6, 1/6 and 6/6, and onset of estrus in 6/6, 0/6, 6/6, 6/6 and 6/6 for T1, T2, T3, T4 and T5, respectively. Heifers from T1, T3 and T4 that ovulated from a persistent follicle, showed estrus 37.5 +/- 12.10 h after implant removal and heifers that developed a new wave of ovarian follicular development showed it at 120.28 +/- 22.81 h (P<0.01). Ovulation occurred at 5.92 +/- 1.72 and 2.22 +/- 1.00 days (P<0.01), respectively. Progesterone concentration was <1 ng/ml from days 7 to 15 in T1, T2 and T4; for T3 progesterone concentration was 2.25 +/- 0.50 ng/ml on day 13 and decreased on day 15 to 0.34 +/- 0.12 ng/ml (P<0.01). For T5, progesterone concentration was 1.66 +/- 0.58 ng/ml on day 15. The more desirable results were obtained with T5, in which 100% of heifers had a new wave of ovarian follicular development induced, with onset of estrus and ovulation synchronized in a short time period.     

Effect of dominant follicle persistence on follicular fluid oestradiol and inhibin and on oocyte maturation in heifers

The aim of the present study was to characterize in detail the cytoplasmic and nuclear morphology of cattle oocytes recovered from follicles that are dominant for more than 9 days (with low fertility after ovulation), and to relate morphological changes to intrafollicular markers of follicle health. Beef heifers received prostaglandin F2 alpha and a synthetic progestagen (3 mg Norgestomet) for 2 or 10 days on the first day of dominance of the second dominant follicle (DF2) of the oestrous cycle, to give a 4 day (n = 19; N2) or 12 day (n = 21; N10) duration of dominance of the dominant follicle at ovariectomy 18 h after implant removal and before the predicted gonadotrophin surge. Ultrasound scanning determined emergence of a new wave of follicles in five N10 heifers the day before (n = 1) or day of ovariectomy (n = 4) (N10-NonDom). Dominant follicles from the remaining N10 heifers (N10-Dom) were larger (P < 0.05) on the day of ovariectomy (17.8 +/- 0.6 mm) than those from N2 heifers (13.6 +/- 0.4 mm). The oestradiol:progesterone ratio of follicular fluid from N10-Dom heifers was reduced (21.7 +/- 3.1 versus 34.1 +/- 4.4; P < 0.05), while inhibin A (as measured by immunoradiometric assay) was increased (12.7 +/- 1.0 versus 9.0 +/- 0.7 micrograms ml-1; P < 0.05) compared with N2 heifers. Eleven of twelve N2 oocytes demonstrated nuclear activation without germinal vesicle breakdown, while seven of eight N10-Dom oocytes had undergone germinal vesicle breakdown and had progressed to metaphase I (6/8) or II (1/8). In contrast to N2 oocytes, N10-Dom oocytes showed a larger perivitelline space containing more cumulus cell process endings, vacuoles, irregular vesicles, and more mitochrondia and lipid droplets throughout the ooplasm, yet the degree of cumulus cell expansion and atresia was similar. Thus, final oocyte maturation leading to metaphase I is initiated in most dominant follicles with a dominance period of > 9 days before the gonadotrophin surge and is associated with a reduction in dominant follicle health. However, ovulatory ability is maintained and will lead to the ovulation of aged oocytes, markedly reducing subsequent pregnancy rates.     

Effect of LH or GnRH on the dominant follicle of the first follicular wave in beef heifers

A study was designed to characterise ovarian follicular dynamics in heifers treated with porcine luteinizing hormone (pLH) or gonadotropin releasing hormone (GnRH) on days 3, 6 or 9 (ovulation = day 0), corresponding to the growing, early-static, and late-static phases of the first follicular wave. Following ovulation, 65 beef heifers were assigned, by replicate, to the following seven treatment groups: 25 mg im of pLH on days 3, 6 or 9 (n = 9 per group); 100 microg im of GnRH on days 3, 6 or 9 (n = 9 per group); or controls (no treatment; n = 11). Ovulation occurred within 36 h in 67%, 100% and 67% of heifers treated with pLH and in 89%, 56% and 22% of heifers treated with GnRH on days 3, 6 or 9, respectively (treatment-by-day interaction, P < 0.09). Combined for all treatment days, ovulation rates were 78% and 56% in pLH- and GnRH-treated groups, respectively (P < 0.09). Overall, mean day (+/- SD) of emergence of the second follicular wave in heifers that ovulated was different from that in controls or in heifers that did not ovulate (P < 0.05). Mean (+/- SD) day of emergence of the second wave occurred earlier (day 5.6+/-1.2; P < 0.05) in heifers that ovulated after treatment on day 3 (n = 14) than in controls (day 8.7+/-1.6; n = 11); however, wave emergence in all heifers treated on day 6 (day 8.1+/-0.5; n = 18) did not differ from controls, regardless of whether or not ovulation occurred. In the heifers that ovulated in response to treatment on day 9 (n = 8), the emergence of the second follicular wave was delayed (day 10.9+/-0.4; P < 0.05). The day of emergence of the second wave in the 14 treated heifers that failed to ovulate, irrespective of the day of treatment (day 8.9+/-1.4) did not differ from control heifers. The emergence of the second wave was more synchronous in day 6 heifers (regardless of whether they ovulated) and in day 9 heifers that ovulated compared to control heifers (P < 0.05). Results did not support the hypothesis that the administration of pLH or GnRH at known stages of the follicular wave in cycling heifers would consistently induce ovulation or atresia and, thereby, induce emergence of a new follicular wave at a predictable interval. New wave emergence was induced consistently (1.3 days post-treatment) only in those animals that ovulated in response to treatment. However, 22% of LH-treated heifers and 44% of GnRH-treated heifers failed to ovulate. Treatments did not induce atresia of the dominant follicle or alter the interval to new wave emergence in animals that did not ovulate in response to treatment.     

Relationship between the onset of oestrus, the preovulatory surge in luteinizing hormone and ovulation following oestrous synchronization and superovulation of farmed red deer (Cervus elaphus).

The timing of ovulation relative to the onset of oestrus and the preovulatory surge in luteinizing hormone (LH) was studied in red deer following treatments to synchronize oestrus and induce either a monovulatory or superovulatory response. Mature hinds (n = 36) were allocated randomly to two mating groups (n = 16 + 20), with respective treatments staggered by 4 weeks during the 1990 rut (March-April). Each hind was treated with an intravaginal controlled internal drug releasing (CIDR)-type S device for 14 days. Treatments to induce a monovulatory response included CIDR device alone (treatment A; n = 4 + 8) and additional injection of 200 iu pregnant mares' serum gonadotrophin (PMSG) at device removal (treatment B; n = 4 + 4). Treatments to induce a superovulatory response included injections of 200 iu PMSG and 0.5 units ovine follicle-stimulating hormone (FSH) at about time of removal of CIDR devices (treatment C; n = 4 + 4) and further treatment with gonadotrophin-releasing hormone (GnRH) analogue 18 h after removal of CIDR devices (treatment D; n = 4 + 4). The hinds were run with crayon-harnessed stags from insertion of CIDR devices (12 March or 9 April) and blood samples were taken every second day to determine plasma progesterone. Further blood samples were collected for determination of plasma LH and progesterone via indwelling jugular cannulae every 2 h for 72 h from removal of CIDR devices. Hinds were allocated randomly to an initial ovarian examination by laparoscopy at either 16 or 20 h (A and B), or 12 or 16 h (C and D) after the onset of oestrus, with laparoscopy repeated at intervals of 8 h until either ovulation was recorded (A and B), or for four successive occasions (C and D). All hinds received cloprostenol injections 15 days after device removal. A total of 28 hinds (78%) exhibited oestrus and a preovulatory LH surge, with mean (+/- SEM) times to onset of oestrus of 44.6 +/- 1.0 h (A; n = 7), 37.4 +/- 2.0 h (B; n = 7), 16.3 +/- 1.7 h (C; n = 6) or 14.0 +/- 1.7 h (D; n = 8). Failure to exhibit oestrus or LH surge was most prevalent among hinds in treatment A early in the rut.(ABSTRACT TRUNCATED AT 400 WORDS)