Effect of bovine follicular fluid administration during or after norgestomet treatment on the fate of the proestrus dominant follicle in heifers

The objective was to examine the influence of follicle stimulating hormone (FSH) on the maintenance and ovulation of the proestrus dominant follicle (DF) in cattle. This was investigated by subjecting a proestrus DF, maintained by a single norgestomet (N) implant, to bovine follicular fluid (BFF) injections during N treatment and immediately after its removal. Earlier, we demonstrated that with an insertion of a single N implant the proestrus DF could be maintained for 9 days without affecting its ovulatory capacity. Eighteen cycling Holstein heifers were treated with a N implant at proestrus. The day of implant insertion was designated day 1 of the implant period and the implant was retained for 9 days. Heifers (n = 6 per group) were randomly allocated to receive saline for 4 days from day 5 to day 8 of the implant period and for 4 days from the day of implant removal to day 3 after removal (CONTROL) or BFF from day 5 to day 8 of implant period (BFF-DURING) or BFF from the day of implant removal to day 3 after implant removal (BFF-AFTER). Injections (10 ml) were given i.v. twice daily and the ovaries monitored by ultrasonography daily, throughout and after the implant period. All CONTROL heifers maintained the DF during treatment and ovulated following implant withdrawal. In all BFF-DURING heifers, the BFF injections caused regression of the DF and its disappearance. In three of the BFF-AFTER heifers, BFF injections caused regression of the DF. In the remaining three BFF-AFTER heifers, the DF ovulated. Mean plasma FSH concentrations did not differ (P > 0.05) between the CONTROL and BFF-DURING heifers. However, the mean plasma FSH concentrations were lower in BFF-AFTER heifers compared with CONTROLS (P < 0.05). Mean plasma LH concentrations did not differ among treatment groups (P > 0.05). In summary, BFF treatment caused atresia of the proestrus DF when maintained by N and this was not associated with suppression of circulating FSH. Administration of BFF after implant removal resulted in an equal chance of ovulation or regression of DF. Regression was associated with suppression of FSH and LH.     

Comparison of three methods of acute administration of progesterone on ovarian follicular development and the timing and synchrony of ovulation in Bos indicus heifers

The aim of this study was to induce the formation of a persistent dominant ovarian follicle and to compare the effects of 3 methods of acute administration of P4 on ovarian follicular development and on the timing and synchrony of ovulation. Stage of the estrous cycle was initially synchronized in Bos indicus heifers with a norgestomet implants (3 mg) for 10 d and with an analogue of PGF2 alpha (15 mg) on the first and last day of norgestomet treatment. Eight days after removal of the implants, heifers were randomly assigned to 4 groups. All heifers received a norgestomet implant (Day 0), which was removed 17 d later (Day 17); PGF2 alpha was administered on Days 0 and 4. Heifers in the control group (n = 5) received no other treatment. On Day 10 heifers in Group P4C (n = 5) were treated with a CIDR for 24 h; heifers in Group P4O (n = 5) were administered 100 mg i.m. of P4 in oil, while heifers in Group P4S (n = 5) were administered 100 mg i.m. of P4 in saline/alcohol. Data were analyzed using bootstrap estimates of location (mean) and spread (standard deviation; SD). Compared with the control heifers, day of emergence of the ovulatory follicle was delayed, and age and duration of dominance of the ovulatory follicle were reduced in the P4C and P4O heifers (P < 0.05) but not in the P4S heifers (P > 0.05). In all groups treated with P4 both the mean and variability (SD) in the timing of ovulation did not differ with that of the control group (P > 0.05) but there was less variability in the day of emergence, age, duration of dominance and diameter of the ovulatory follicle than in the control group (P < 0.05). Delayed timing and reduced synchrony (SD) of ovulation and greater age of the ovulatory follicle (P < 0.05) occurred in P4S heifers than in P4C heifers. We conclude that administration of 100 mg of P4 in oil is as effective as treatment with a CIDR for synchronizing emergence and ovulation of a newly recruited dominant follicle. However, reduced synchrony of ovulation, greater age of the ovulatory follicle and delayed timing of ovulation occurred following administration 100 mg of P4 in saline/alcohol compared with the CIDR device.     

Influence of perioestrous suprabasal progesterone levels on cycle length,oestrous behaviour and ovulation in heifers

In order to induce suprabasal plasma concentrations of progesterone after luteolysis and to determine their effect on oestrous behaviour and ovulation, heifers subcutaneously received silicone implants containing 2.5 (n = 4), 5 (n = 4), 6 (n = 3), 7.5 (n = 3) or 10 (n = 4) g of progesterone, or an empty implant (controls, n = 5) between days 8 and 25 of the cycle (ovulation designated Day 0). Growth of dominant follicles and time of ovulation were determined by ultrasound, and signs of oestrus were recorded and scored. Blood was collected at 2–4 h intervals from Days 15 to 27 and assayed for progesterone concentration. In all heifers, plasma concentrations of progesterone sharply decreased during Days 16–18. Control heifers had their lowest progesterone levels on Days 20.5 and 21, standing oestrus on Day 19.5 ± 0.4 (mean ± SEM), and ovulated on Day 20.7 ± 0.4. A similar pattern was observed in heifers treated with 2.5 and 5 g progesterone. Heifers treated with 6, 7.5 and 10 g of progesterone showed an extended (P < 0.05) interovulatory interval. Onset of prooestrus and time of maximum expression of signs of oestrus were not significantly different from those in controls. However, there was an absence of standing oestrus in most of the cases, signs of oestrus lasted longer (P < 0.05) and were weaker in intensity when doses increased. In these groups, the lowest progesterone concentrations were attained shortly after implant removal. Some heifers treated with 6 and 7.5 g of progesterone had standing oestrus and post oestrous bleeding as seen in the controls but ovulation occurred from Days 24.5 to 27. When plasma progesterone concentrations were over 1 nmol 1⁻¹, disturbed oestrus and delayed ovulation occurred. The extended period of prooestrus and oestrus and delayed ovulation were similar to that described in cases of repeat breeding. It is suggested that suprabasal plasma concentrations of progesterone, after luteolysis, may lead to asynchrony between onset of oestrus and ovulation and consequently be a cause of repeat breeding in cattle.     

Follicular dynamics and temporal relationships among body temperature, oestrus, the surge of luteinizing hormone and ovulation in Holstein heifers treated with norgestomet

The effects of chronic treatment with norgestomet on follicular dynamics, corpus luteum growth and function as well as the temporal relationships among body temperature, oestrous behaviour, the luteinizing hormone (LH) surge and ovulation following implant removal were studied in 16 Holstein heifers. Oestrous cycles of the heifers were initially synchronized using 2 injections of prostaglandin F-2 alpha (PGF-2 alpha) 12 days apart. The heifers were then implanted with a norgestomet ear implant for 9 days, beginning either at the middle of the synchronized cycle (dioestrus) or at the end of the synchronized cycle (pro-oestrus). Follicular dynamics, corpus luteum growth and regression, and plasma progesterone were not affected by norgestomet treatment at dioestrus. The dominant follicle present at the time of norgestomet implantation in the pro-oestrus group was maintained during the 9-day implant period of 6 of 8 heifers and ovulated after implant removal. Time from implant removal to onset of standing oestrus and time to LH peak following implant removal were highly correlated with the time of ovulation (r = 0.92 and 0.96, respectively). Onset of standing oestrus and the LH peak and the onset of standing oestrus and peak vaginal and rectal temperatures were also highly correlated (r = 0.96, 0.82 and 0.81, respectively). It is concluded that any decrease in pregnancy rates following treatment with norgestomet is not due to asynchrony among oestrus, the LH surge and ovulation.     

Comparison of long-term CIDR-based protocols to synchronize estrus in beef heifers

Two experiments evaluated long-term controlled internal drug release (CIDR) insert-based protocols to synchronize estrus and compare differences in their potential ability to facilitate fixed-time artificial insemination (FTAI) in beef heifers. In Experiment 1 estrous cycling heifers (n = 85) were assigned to one of two treatments by age and body weight (BW). Heifers with T1 received a CIDR from days 0 to 14, gonadotropin releasing hormone (GnRH) on day 23, and prostaglandin F_2α (PG) on day 30. Heifers with T2 received a CIDR from days 2 to 16, GnRH on day 23, and PG on day 30. Ovaries were evaluated by ultrasonography on days 23 and 25 to determine ovulatory response to GnRH. In Experiment 2 heifers (n = 353) were assigned within reproductive tract scores by age and BW to one of four treatments. Heifers in T1 and T2 received the same treatments described in Experiment 1. Heifers in T3 and T4 received the same treatments as T1 and T2, respectively, minus the addition of GnRH. In Experiments 1 and 2, heifers were fitted with HeatWatch transmitters for estrous detection and AI was performed 12 h after estrus. In Experiment 1 heifers assigned to T1 had larger dominant follicles at GnRH compared to T2 (P < 0.01) but response to GnRH, estrous response after PG, mean interval to estrus, and variance for interval to estrus after PG did not differ (P > 0.10). AI conception and final pregnancy rate were similar (P > 0.50). In Experiment 2 estrous response after PG did not differ (P > 0.70). Differences in mean interval to estrus and variance for interval to estrus (P < 0.05) differed based on the three-way interaction of treatment length, GnRH, and estrous cyclicity status. AI conception and final pregnancy rates were similar (P > 0.10). In summary, the greater estrous response following PG and resulting AI conception and final pregnancy rates reported for heifers assigned to the two treatments in Experiment 1 and among the four treatments in Experiment 2 suggest that each of these long-term CIDR-based protocols was effective in synchronizing estrus in prepubertal and estrous cycling beef heifers. However, the three-way interaction involving treatment length, GnRH, and estrous cyclicity status in Experiment 2 clearly suggests that further evaluation of long-term CIDR-based protocols is required with and without the addition of GnRH and on the basis of estrous cyclicity status to determine the efficacy of these protocols for use in facilitating FTAI.     

Fertility of sows injected with exogenous oestradiol and/or gonadotrophins to control post-weaning oestrus

In the first of two experiments 28 multiparous sows were allocated to one of the following treatments 2 days after weaning at approximately 35 days post partum: (1) untreated; (2) i.m. injection 10 μg oestradiol benzoate (OB)/kg body weight (b.wt.); and (3) i.m. injection 20 μg OB/kg b.wt. Sows were bred at first post-weaning oestrus and ovulation rate assessed at slaughter. The mean interval from weaning to oestrus in each group was: (1) 5.6 ± 0.2; (2) 4.7 ± 0.2; and (3) 4.7 ± 0.2 days; the mean ovulation rates in groups 1 and 2 (18.7 ± 0.6 and 17.4 ± 1.8, respectively) were significantly higher (P < 0.01) than that of 12.0 ± 1.7 for treatment 3 sows. Two untreated and one each of the treated sows were not cycling at slaughter.In the second experiment 75 multiparous sows weaned at 28 ± 3 days post partum (day 0) were evenly allocated with respect to parity to one of four treatment groups: (1) untreated; (2) i.m. injection 10 μg OB/kg b.wt. on day 2; (3) PG600 (400 iu PMSG + 200 iu hCG) injection subcutaneous day 0; and (4) combined PG600/OB treatment as in (2) and (3) above. Sows were bred naturally at the first post-weaning oestrus and fertility assessed at farrowing. Control animals had a significantly longer (P < 0.05) weaning to oestrus interval (4.53 ± 0.25 days) compared to treatment 2 (4.03 ± 0.13) treatment 3 (3.97 ± 0.12) and treatment 4 (3.81 ± 0.07) sows. Sows treated with PG600 alone showed a significant increase (P < 0.05) in numbers born live compared to pre-treatment values. A smaller and non-significant increase in numbers born live in control sows (probably related to increasing parity) was not observed in either OB- or PG600/OB-treated animals.These results suggest that with further modification of the treatments, a system may be developed for introducing fixed-time artificial insemination (AI) or mating as a means of controlling the reproductive performance of the weaned sow.     

Reproductive performance of heifers induced to oestrous asynchrony by suprabasal plasma progesterone levels

Suprabasal progesterone concentrations around oestrus have induced disturbances in oestrous behaviour and ovulation. To determine whether fertility in such an altered oestrus can be maintained at normal levels with additional inseminations (AI) until ovulation, fertility was compared in heifers (n = 11) inseminated in normal oestrous cycles and thereafter in cycles in which the animals were treated with progesterone in order to create suprabasal concentrations after luteolysis. The treatment consisted of silicone implants containing 10.6 mg kg⁻¹ of progesterone inserted subcutaneously on Day 8 of the oestrous cycle (day of ovulation designated Day 0) and removed on Day 25. Both in control oestrous cycles and oestrous cycles under progesterone treatment, growth of the ovulatory follicle and ovulation were determined by frequent ultrasound scanning. Blood was collected frequently for further analysis of progesterone, oestradiol-17β and luteinising hormone (LH). Insemination was performed 12 h after onset of standing oestrus. if ovulation did not occur 24 h after AI, heifers were inseminated again until ovulation. Pregnancy was diagnosed by ultrasound 25 days after ovulation.In control oestrous cycles, plasma progesterone decreased to 0.3 ± 0.3 nmol 1⁻¹. Duration of oestrus was 22.9 ± 2.0 h, the interval from onset of oestrus to ovulation was 32.4 ± 2.3 h and the interval from LH peak to ovulation was 28.6 ± 1.4 h. The interovulatory interval was 20.7 ± 0.6 days. In oestrous cycles in treated heifers, progesterone decreased to 1.0 ± 0.3 nmol l⁻¹ (P > 0.10) and the interovulatory interval was prolonged to 23.5 ± 1.0 days (P < 0.05). Standing oestrus lasted 47.2 ± 12.0 h (P = 0.09, n = 7). The interval from the onset of oestrus to ovulation was 59.4 ± 13.0 h (P = 0.08) and the interval from LH peak to ovulation 25.8 ± 1.3 h (P > 0.10). The prolonged oestrus was associated with increased (P < 0.05) growth of the ovulatory follicle and higher (P < 0.05) release of oestradiol-17β. Conception rates were 90% and 46% (P < 0.05), and the numbers of AI per heifer were 1.1 ± 0.1 and 3.4 ± 0.6 (P < 0.01) for control oestrous cycles and after treatment, respectively.The induction of suprabasal concentrations of progesterone caused asynchronies similar to those observed in cases of repeat breeding. The repeated AI did not maintain fertility at normal levels. It is suggested that the extended growth of the ovulatory follicle may cause impaired oocyte maturation or it may alter the maternal milieu owing to the prolonged release of oestradiol.     

Superovulatory response following ablation-induced follicular wave emergence at random stages of the oestrous cycle in cattle

Based on the premise that superovulation in cattle is optimal when superstimulation is initiated at the time of follicular wave emergence, the present study was done in beef heifers to determine if the superovulatory response following a single bolus of gonadotrophin treatment after follicle ablation (induced wave) at random stages of the oestrous cycle is comparable to the same gonadotrophin treatment at mid-dioestrus (spontaneous wave). In Experiment 1, heifers were assigned to nonablation (n = 18) and ablation (n = 20) groups. In nonablated heifers, superstimulatory treatment was given as a single subcutaneous injection (Folltropin-V, 400 mg) at mid-dioestrus to coincide with emergence of the spontaneous follicular wave 8 to 12 days after oestrus. In ablated heifers, the same superstimulatory treatment was given 1 day after ablation of all follicles ≥ 5 mm at random stages of the oestrous cycle to coincide with emergence of the ablation-induced wave. In both the nonablation and ablation groups, PGF_2α (Estrumate, 500 μg) was given 48 h after the superstimulatory treatment and artificial insemination was done 60 and 72 h later. Reproductive tracts were collected at the time of slaughter 6 or 7 days after insemination. Observations made in Experiment 1, indicated that some ablated heifers had only partial luteal regression at the time of insemination, while some others exhibited behavioral oestrus as early as 24 h after PGF_2α treatment. The design was amended in Experiment 2 to address these problems. Heifers were assigned to nonablation (n = 17), ablation-alone (n = 20) or ablation plus progestogen (n = 20) groups. Follicle ablation, superstimulatory treatment, artificial insemination and collection of reproductive tracts were done as in Experiment 1. However, all heifers were given two doses of PGF_2α (500 μg/dose) 48 and 60 h after superstimulatory treatment to ensure complete luteal regression, and heifers in the ablation plus progestogen group received a norgestomet ear implant at the time of follicle ablation to prevent early ovulations. The implant was removed at the time of the second PGF_2α treatment. In Experiments 1 and 2, the means for the ovarian and superovulatory responses were not significantly different between groups. Averaged over the nonablation and all ablation groups for Experiments 1 and 2, the mean number of corpora lutea, fertilized ova and transferable embryos were 22.9 vs 18.6, 7.3 vs 7.8 and 5.4 vs 5.6, respectively. In summary, follicle ablation at random stages of the oestrous cycle followed by a single bolus of gonadotrophin treatment 1 day later resulted in a superovulatory response that was comparable to the same superstimulatory treatment administered around the time of spontaneous wave emergence at mid-dioestrus. The ablation/superstimulation method described herein offers the advantage of initiating superstimulatory treatment forthwith and assuring that treatment is concomitant with wave emergence to achieve an optimal superovulatory response. Moreover, the full extent of the oestrous cycle is available for superstimulation and the need for detecting oestrus or ovulation and waiting 8 to 12 days to initiate treatment is eliminated.     

The effect of a progesterone (P4) intravaginal device (CIDR) on resynchronisation of oestrus and embryonic loss in previously timed inseminated dairy heifers

A study was done to evaluate the effect of using progesterone (P4) intravaginal device (CIDR: controlled internal drug-releasing dispenser) to synchronise the return to oestrus of previously timed inseminated (TAI) dairy heifers, and to evaluate embryo survival and pregnancy rate (PR) in the return to oestrus heifers. At the onset of the artificial insemination (AI) breeding period (day -9), heifers were randomly assigned into two groups (treated group CGPG, n = 79) and (control group GPG, n = 83). Every heifer in both groups was injected with gonadotropin-releasing hormone (GnRH) agonist and prostaglandin F2-alpha (PGF2α) as follows: GnRH on day -9; PGF2α on day -2; GnRH and TAI on day 0. Heifers in both groups received TAI within 30 min after the second GnRH injection. Artificial insemination at first breeding was conducted for all heifers during 55 days from day 0. On day 14 after timed insemination, every heifer in the CGPG group received CIDR device for 6 days. Within 3 days after CIDR removal, more heifers in CGPG group showed oestrus within 1.9 days compared to heifers that showed oestrus within 2.9 days in the control. Within 10 days after CIDR removal, more heifers in the CGPG group showed oestrus within 2.4 days compared to heifers that showed oestrus within 6.7 days in the control. PRs on days 30 and 55 were not different between both groups, while PR on day 55 during September were higher (P = 0.032) in CGPG group (58.0%) than GPG group (37.0%). In addition, PR from first to second AI was higher (P = 0.037) for CGPG group (79.8%) than for GPG group (65.1%) but it was similar after that. Pregnancy losses between days 30 and 55 tended to be lower (P = 0.089) for the CGPG group (12.7%) compared to 25.1% for the GPG group. Interval between first and second AI was lower (P = 0.052) for the CGPG group (27.5 ± 1.6 days) compared to 31.6 ± 1.3 days for heifers in the GPG group but no differences were detected for intervals from second to third AI and from third to fourth AI between the two groups. Number of services per pregnancy was not different between CGPG and GPG groups. Results indicate that the CIDR device improved synchronisation to return to oestrus and increased PR to first AI during high temperature months by reducing embryonic losses.     

Variable progesterone response and estradiol secretion in prepubertal beef heifers following treatment with norgestomet implants

Two experiments were conducted to determine the effects of norgestomet ear implants on progesterone response and estradiol secretion in prepubertal beef heifers. In the first experiment, 47 beef heifers were treated with norgestomet. The implants were implanted subcutaneously for 9 d. After implant removal, blood samples were taken from heifers 2 to 4 d per week for 40 d. Following progesterone determination in jugular venous plasma, heifers were classified according to their progesterone response: 1) no response (Group 1); no rise in progesterone above 1 ng/ml throughout the sampling period; 2) one cycle (Group 2); one increase in progesterone above 1 ng/ml for at least 2 d followed by no further increase in progesterone during the sampling period; and 3) two cycles (Group 3); a rise in progesterone above 1 ng/ml for at least 2 d followed by another cycle of normal duration. Heifers treated with norgestomet were classified as 23 with no response, 9 with 1 cycle and 15 with 2 cycles. Concentrations of estradiol were measured in jugular venous samples on Day 2 after implant removal. Mean concentrations of estradiol were greater in Group 3 than in Group 1 (P < or = 0.01). In Experiment 2, 29 prepubertal beef heifers were assigned randomly to either a 9-d treatment with norgestomet (n = 14) or to serve as untreated controls (n = 15). Blood plasma samples were collected daily from Days 0 to 44 after implant removal. After progesterone determination, heifers were classified as 8 with no response, 4 with 1 cycle and 3 with 2 cycles in the control group, and 5 with no response, 3 with 1 cycle and 6 with 2 cycles in the norgestomet group (frequencies did not differ; P > 0.1). Jugular venous blood plasma was also collected at 4-h intervals from 0 h to 96 h after implant removal and concentrations of estradiol were measured. Patterns of estradiol secretion differed (P < or = 0.05) and overall mean concentrations of estradiol over the first 96 h following implant removal were greater (P < or = 0.01) in norgestomet-treated heifers versus the controls. We conclude that norgestomet can produce a variable progesterone response with heifers with 2 cycles secreting more estradiol. Implants of norgestomet also causes more acute secretion of estradiol in prepubertal beef heifers.     

Effect of chronic treatment with bromocryptine on the corpus luteum function of the cow

Six heifers received an intramuscular injection of 15 mg bromocryptine twice daily from day 1 (the day of standing oestrus was defined as day 0) until 50 h after the start of luteal regression. The overall mean level of prolactin was 0.22 ±0.01 μg/l (SEM; n=6) in the bromocryptine-treated group and 10.7±2.7 μg/1 (SEM; n=6) in the control group. No significant differences in the overall mean level of progesterone and LH, the mean length of the early-luteal phase, the luteal phase and the period of luteal regression were measured between the two groups. The results provide strong evidence that prolactin has no luteotrophic properties in the cow during the oestrous cycle.     

Estrus after treatment with Syncro-Mate B* in ovariectomized heifers is dependent on the injected estradiol valerate

A series of experiments was conducted to determine why ovariectomized heifers exhibit estrus after they are treated with the estrus synchronization product, Syncro-Mate B(*) (SMB). In Experiment 1, 23 of 40 (58%) ovariectomized heifers exhibited estrus after treatment with SMB. The mean concentration of estradiol-17beta (E(2)) in serum was lower (P < 0.001) before treatment than after implant removal in ovariectomized heifers treated with SMB. Six of 10 heifers from which serum was collected to determine concentrations of LH exhibited estrus and 5 of 6 had a surge of LH in serum after implant removal. In Experiment 2, when no estradiol valerate (EV) was given or when the norgestomet implant period was extended from 9 to 18 d, no heifer exhibited estrus after implant removal. The mean concentration of E(2) for 3 d after implant removal was lower (P < 0.001) in ovariectomized heifers with implants for 18 d versus those with implants for 9 d and was also lower (P < 0.001) in ovariectomized heifers treated only with norgestomet compared with those receiving the standard SMB treatment. When estradiol-17beta was substituted for EV in the SMB treatment, serum E(2) was lower (P < 0.001) after implant removal than in heifers receiving the standard SMB treatment. Experiment 3 demonstrated that combining a norgestomet implant or implant plus a 3-mg injection of norgestomet with EV did not alter concentrations of E(2) in serum on the days when synchronized estrus would be expected following SMB treatment. The results indicate that the SMB-induced estrus in ovariectomized heifers is dependent upon EV in the SMB treatment. Apparently, EV elevates the concentration of E(2) in serum, and the E(2) remains sufficiently high to induce estrus after implant removal.     

Effects of repetitive norgestomet treatments on pregnancy rates in cyclic and anestrous beef heifers

Sixty-eight 12- to 14-month-old crossbred beef heifers averaging 285 kg were assigned at random to treated (n = 35) and control (n = 33) groups to evaluate the use of repetitive norgestomet treatments. Treated heifers received an ear implant containing 6 mg norgestomet on two occasions 16 days apart. Injections of 3 mg norgestomet and 5 mg estradiol valerate (EV) were given intramuscularly the same day as first implantation (Syncro-Mate-B). Implants were removed after eight days. Four bulls were then placed in each of two pastures containing half of the treated and half of the control heifers for 24 days after the time of the first implant removal. Progesterone concentrations from blood samples collected prior to the first treatment were used to determine reproductive status. The overall pregnancy rate 64 days after first implant removal for treated anestrous heifers (61%; 14 23 ) was similar (p > 0.25) to untreated (73%; 11 15 ) and treated (75%; 9 12 ) cyclic heifers, but higher (p < 0.1) than for untreated anestrous heifers (33%; 6 18 ). This treatment advantage resulted from an increased (p < 0.01) pregnancy rate after the second implant removal. In summary, repetitive norgestomet treatments enhanced pregnancy rate in anestrous heifers within a 24-day breeding season.     

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.     

Normal or induced secretory patterns of luteinising hormone and follicle-stimulating hormone in anoestrous gonadotrophin-releasing hormone-immunised and cyclic control heifers

The objective was to determine the effect of gonadotrophin-releasing hormone (GnRH), GnRH analogue (GnRH-A) or oestradiol administration on luteinising hormone (LH) and follicle-stimulating hormone (FSH) release in GnRH-immunised anoestrous and control cyclic heifers. Thirty-two heifers (477 ± 7.1 kg) were immunised against either human serum albumin (HSA; controls; n = 8), or a HSAGnRH conjugate. On day 70 after primary immunisation, control heifers (n = 4 per treatment; day 3 of cycle) received either (a) 2.5 μg GnRH or (b) 2.5 μg of GnRH-A (Buserelin®) and GnRH-immunised heifers (blocked by GnRH antibody titre; n = 6 per treatment) received either (c) saline, (d) 2.5 μg GnRH, (e) 25 μg GnRH or (f) 2.5 μg GnRH-A, intravenously. On day 105, 1 mg oestradiol was injected (intramuscularly) into control (n = 6) and GnRH-immunised anoestrous heifers with either low (13.4 ± 1.9% binding at 1:640; n = 6) or high GnRH antibody titres (33.4 ± 4.8% binding; n = 6). Data were analysed by ANOVA. Mean plasma LH and FSH concentrations on day 69 were higher (P < 0.05) in control than in GnRH-immunised heifers (3.1 ± 0.16 vs. 2.5 ± 0.12 ng LH ml⁻¹ and 22.5 ± 0.73 vs. 17.1 ± 0.64 ng FSH ml⁻¹, respectively). The number of LH pulses was higher (P < 0.05) in control than in GnRH-immunised heifers on day 69 (3.4 ± 0.45 and 1.0 ± 0.26 pulses per 6 h, respectively). On day 70, 2.5 μg GnRH increased (P < 0.05) LH concentrations in control but not in GnRH-immunised heifers, while both 25 μg GnRH and 2.5 μg GnRH-A increased (P < 0.05) LH concentrations in GnRH-immunised heifers, and 2.5 μg GnRH-A increased LH in controls. FSH was increased (P < 0.05) in GnRH-immunised heifers following 25 μg GnRH and 2.5 μg GnRH-A. Oestradiol challenge increased (P < 0.05) LH concentrations during the 13–24 h period after challenge with a greater (P < 0.05) increase in control than in GnRH-immunised heifers. FSH concentrations were decreased (P < 0.05) for at least 30 h after oestradiol challenge. In conclusion, GnRH immunisation decreased LH pulsatility and mean LH and FSH concentrations. GnRH antibodies neutralised low doses of GnRH (2.5 μg), but not high doses of GnRH (25 μg) and GnRH-A (2.5 μg). GnRH immunisation decreased the rise in LH concentrations following oestradiol challenge.     

Peripheral progesterone levels in pregnant and non-pregnant heifers following use of HCG.

Progesterone concentration in jugular blood of bred beef heifers was determined on days 9 and 16 in two trials. Human chorionic gonadotrophin (HCG) was administered to some of the heifers in each trial in an attempt to improve pregnancy percentage.In Trial 1, 183 heifers were divided into a control group and three groups of animals which were subjected to a 9-day estrous synchronization treatment prior to breeding. The treatment consisted of an ear implant containing 17 α acetoxy-11-beta-methyl 17 nor preg 4-ene 3, 20 dione (norgestomet) left in place for 9 days and an injection of 5 mg estradiol valerate (EV) and 3 mg of norgestomet given at the time of implantation. The heifers in one group received .25 mg estradiol-17β at time of implant removal; heifers in the 2nd group received 1500 IU of HCG in 5% beeswax and 95% sesame oil at breeding time, while heifers in the 3rd group received a placebo injection containing 5% beeswax and 95% sesame oil at breeding time. No differences in serum levels of progesterone were observed (P>0.5) between treatments or between pregnant and non-pregnant heifers on day 9 or 16 (P>.05). Pregnancy percentage in heifers receiving HCG was similar to that noted in the control heifers or the placebo injected heifers while injection of estradiol 17β decreased the proportion of heifers which became pregnant.In trial 2, 58 heifers which had been bred 1 or 2 times without becoming pregnant were divided into a control group and a group in which heifers received 1000 IU of HCG 96 hr. after observed estrus. In heifers receiving HCG, serum levels of progesterone were higher (P<.01), on day 9 and 16 post estrus than in controls but no difference in serum progesterone was noted (P>.05) between pregnant and non-pregnant heifers on day 9 or 16. The proportion pregnant did not differ (P>.05) between heifers receiving HCG and control heifers.     

GnRH treatment at artificial insemination in beef cattle fails to increase plasma progesterone concentrations or pregnancy rates

Treatment with GnRH at the onset of standing estrus increased pregnancy percentages and circulating concentrations of progesterone in repeat breeder dairy cows. The objective of this study was to determine the effect of treatment with GnRH at AI on concentrations of progesterone and conception rates in beef cattle that exhibited estrus. Two hundred ninety-three heifers at four locations were synchronized with the Select Synch plus CIDR protocol (given GnRH and a CIDR was placed into the vagina, and 7 d later, given PGF_2α and CIDR removed; n = 253) or the 14-19 melengestrol acetate (MGA) protocol (MGA fed at 0.5 mg/head/d for 14 d, with PGF_2α 19 d after MGA withdrawal n = 40) and AI was done after detection of estrus. At Location 1, blood samples were collected on Day 2, 4, 6, 10, 15, and 18 after AI (Day 0 = AI). Two hundred and fifty postpartum cows at two locations were synchronized with the Select Synch plus CIDR protocol, and AI was performed after detection of estrus. At AI, cattle were alternately assigned to one of two treatments: (1) treatment with GnRH (100 μg) at AI (n = 127 heifers and n = 108 cows); or (2) non-treated control (n = 120 heifers and n = 119 cows). Concentrations of progesterone tended to be greater in control heifers compared to GnRH-treated heifers on Days 6 (P = 0.08), 10 (P = 0.07), and 15 (P = 0.11). Overall conception rates were 68% and 66% for GnRH treated and control, respectively, and were not different between treatments (P = 0.72). In summary, treatment with GnRH at time of AI had no influence on conception rates in cattle that had exhibited estrus.     

Effect of progesterone concentrations,follicle diameter,timing of artificial insemination,and ovulatory stimulus on pregnancy rate to synchronized artificial insemination in postpubertal Nellore heifers

Two experiments were designed to evaluate the effects of treatments with low versus high serum progesterone (P₄) concentrations on factors associated with pregnancy success in postpubertal Nellore heifers submitted to either conventional or fixed timed artificial insemination (FTAI). Heifers were synchronized with a new controlled internal drug release device (CIDR; 1.9 g of P₄ [CIDR1]) or a CIDR previously used for 18 days (CIDR3) plus 2 mg of estradiol (E₂) benzoate on Day 0 and 12.5 mg of prostaglandin F2α on Day 7. In experiment 1 (n = 723), CIDR were removed on Day 7 or 9 and heifers were inseminated after estrus detection. In experiment 2 (n = 1083), CIDR were all removed on Day 9 and FTAI was performed either 48 hours later in heifers that received E₂ cypionate (ECP) on Day 9 (0.5 mg; E48) or 54 or 72 hours later in conjunction with administration of GnRH (100 μg; G54 or G72). Synchronization with CIDR1 resulted in greater serum P₄ concentrations and smaller follicle diameters on Days 7 and 9 in both experiments. In experiment 1, treatment with CIDR for 9 days decreased the interval from CIDR removal to estrus (Day 7, 3.76 ± 0.08 days vs. Day 9, 2.90 ± 0.07; P < 0.01) and improved conception (Day 7, 57.1% vs. Day 9, 65.8%; P = 0.05) and pregnancy rates (Day 7, 37.6% vs. Day 9, 45.3%; P = 0.04). In experiment 2, treatment with ECP improved (P < 0.01) the proportion of heifers in estrus (E48, 40.9%^a; G54, 17.1%^c; and G72, 32.0%^b), but the pregnancy rate was not affected (P = 0.64) by treatments (E48, 38.8%; G54, 35.5%; G72, 37.5%). Synchronization with CIDR3 increased follicle diameter at FTAI (CIDR1, 11.07 ± 0.10 vs. CIDR3, 11.61 ± 0.10 mm; P < 0.01), ovulation rate (CIDR1, 82.8% vs. CIDR3, 88.0%; P < 0.01) and did not affect conception (CIDR1, 42.2 vs. CIDR3, 45.1%; P = 0.38) or pregnancy rates (CIDR1, 34.7 vs. CIDR3, 39.4%; P = 0.11). In conclusion, length of treatment with P₄ affected the fertility of heifers bred based on estrus detection. When the heifers were submitted to FTAI protocol, follicle diameter at FTAI (≤10.7 mm, 23.6%; 10.8–15.7 mm, 51.5%; ≥15.8 mm, 30.0%; P < 0.01) was the main factor that affected conception and pregnancy rates.     

Effect of estradiol valerate on ovarian follicle dynamics and superovulatory response in progestin-treated cattle

Three experiments evaluated the effects of estradiol valerate (EV) on ovarian follicular and CL dynamics, intervals to estrus and ovulation, and superovulatory response in cattle. Experiment 1 compared the efficacy of two norgestomet ear implants (Crestar and Syncro-Mate B; SMB) for 9 d (with PGF at implant removal), combined with either 5 mg estradiol-17beta and 100 mg progesterone (EP) or 5 mg EV and 3mg norgestomet (EN) im at the time of implant insertion on CL diameter and follicular wave dynamics. Ovaries were monitored by ultrasonography. There was no effect of norgestomet implant. Diameter of the CL decreased following EN treatment (P < 0.01). Mean (+/- S.D.) day of follicular wave emergence (FWE) was earlier (P < 0.0001) and less variable (P < 0.0001) in EP- (3.6 +/- 0.5 d) than in EN- (5.7 +/- 1.5 d) treated heifers. Intervals from implant removal to estrus (P < 0.001) and ovulation (P < 0.01) were shorter in EN- (45.7 +/- 11.7 and 74.3 +/- 12.6 h, respectively) than in EP- (56.4 +/- 14.1 and 83.3 +/- 17.0 h, respectively) treated heifers. Experiment 2 compared the efficacy of EP versus EN in synchronizing FWE for superovulation in SMB-implanted cows. At random stages of the estrous cycle, Holstein cows (n = 78) received two SMB implants (Day 0) and were randomly assigned to receive EN on Day 0 or EP on Day 1. Folltropin-V treatments were initiated on the evening of Day 5, with PGF in the morning and evening of Day 8, when SMB were removed. Cows were inseminated after the onset of estrus and embryos were recovered 7 d later. Non-lactating cows had more CL (16.7 +/- 11.3 versus 8.3 +/- 4.9) and total ova/embryos (14.7 +/- 9.5 versus 7.9 +/- 4.6) than lactating cows (P < 0.05). EP-treated cows tended (P = 0.09) to yield more transferable embryos (5.6 +/- 5.2) than EN-treated cows (4.0 +/- 3.7). Experiment 3 compared the effect of dose of EV on ovarian follicle and CL growth profiles and synchrony of estrus and ovulation in CIDR-treated beef cows (n = 43). At random stages of the estrous cycle (Day 0), cows received a CIDR and no further treatment (Control), or an injection of 1, 2, or 5 mg im of EV. On Day 7, CIDR were removed and cows received PGF. Follicular wave emergence occurred within 7 d in 7/10 Control cows and 31/32 EV-treated cows (P < 0.05). In responding cows, interval from treatment to FWE was longer (P < 0.05) in those treated with 5 mg EV (4.8 +/- 1.2 d) than in those treated with 1 mg (3.2 +/- 0.9 d) or 2 mg (3.4 +/- 0.8 d) EV, while Control cows were intermediate (3.8 +/- 2.0 d). Diameter of the dominant follicle was smaller (P < 0.05) at CIDR removal and tended (P = 0.08) to be smaller just prior to ovulation in the 5 mg EV group (8.5 +/- 2.2 and 13.2 +/- 0.6 mm, respectively) than in the Control (11.8 +/- 4.6 and 15.5 +/- 2.9 mm, respectively) or 1mg EV (11.7 +/- 2.5 and 15.1 +/- 2.2 mm, respectively) groups, with the 2mg EV group (10.7 +/- 1.5 and 14.3 +/- 1.7 mm, respectively) intermediate. Diameter of the dominant follicle at CIDR removal was less variable (P < 0.01) in the 2 and 5mg EV groups than in the Control group, and intermediate in the 1mg EV group. In summary, treatment with 5mg EV resulted in a longer and more variable interval to follicular wave emergence than treatment with 5mg estradiol-17beta, which affected preovulatory dominant follicle size following progestin removal, and may have also affected superstimulatory response in Holstein cows. Additionally, 5 mg EV appeared to induce luteolysis in heifers, reducing the interval to ovulation following norgestomet removal. Conversely, intervals to, and synchrony of, follicular wave emergence, estrus and ovulation following treatment with 1 or 2 mg EV suggested that reduced doses of EV may be more useful for the synchronization of follicular wave emergence in progestogen-treated cattle.     

Effects of short-term oilseed supplementation on plasma fatty acid composition,progesterone and prostaglandin F metabolite in lactating beef cows

Twenty-four 3-year-old Angus cows (512.2±21.6 kg) and six ruminally cannulated beef heifers (523.1±16.9 kg) were used to determine the impact of feeding oilseeds starting at the beginning of estrous synchronization until maternal recognition of pregnancy on plasma fatty acid composition. Starting ~60 days postpartum cows were synchronized with the Select Synch+controlled internal drug-release (CIDR) device and timed artificial insemination (AI) protocol. The day CIDR was inserted; cattle were randomly assigned to one of the three treatments being grazing only (CON) or a supplement containing whole soybeans (SOY); or whole flaxseed (FLX). Cattle continued to receive these diets for 28 days. Blood was collected every 3 days until 10 days after insemination and then every day until 18 days after insemination. All cattle grazed a common pasture and supplemented cattle were individually fed their respective supplements once daily. Ruminally cannulated heifers were used to evaluate the impact supplements had on forage intake, which was reduced (P=0.05) with oilseed supplementation. Feeding oilseeds increased total fatty acid intake (P<0.001) across treatments with SOY having greater (P<0.001) 18:2n-6 intake than either CON or FLX. Likewise, cattle fed FLX had greater (P<0.001) 18:3n-3 intake than either CON or SOY. There was a treatment×time interaction (P⩽0.05) for all fatty acids identified except for 20:5n-3 (P=0.99). Within 3 days after the start of supplementation, plasma concentrations of 18:2n-6 increased (P<0.001) for cattle fed SOY compared with CON or FLX, whereas flax-fed cattle did not exhibit an increase (P=0.02) until day 15 of supplementation over that of CON. Plasma concentrations for 18:3n-3 was greater (P<0.013) for FLX than both CON and SOY by day 12. Feeding flaxseed tended to (P=0.07) increase and increased (P=0.01) plasma concentrations of 20:4n-6 by day 18 over CON and SOY, respectively. Overall, treatment did not affect serum concentration of progesterone (P=0.18) or prostaglandin F metabolite (P=0.89). However, day after breeding had an effect on serum progesterone (P=0.01) with day 16 after timed AI being lower compared with other days. Feeding oilseeds during the time of estrous synchronization will not only increase the energy density of the diet but will provide key fatty acids around the time of maternal recognition of pregnancy.